Due to recent declines regarding pollinators, The Swedish Civil Contingencies Agency launched a project investigating possible biological threats towards wild bumblebees. This led to an extensive collection of bumblebees from areas in southern Sweden during the summer of 2018. The purpose of this project was to investigate the prevalence of ten bumblebee pathogens and see if there were any difference in prevalence in areas with and areas without commercial bumblebees. This study also aimed to investigate if the prevalence of the pathogens tended to fluctuate during the season. Of the collected bumblebees (Bombus spp) the ones from the late summer season were analyzed in this project, and data from the already analyzed samples from the early summer season was used. To screen the bumblebees for the pathogens a laboratory process was followed with dissections – homogenization – DNA and RNA extractions – conversion of RNA to cDNA and lastly real-time quantitative PCR. The results showed significant differences between areas with and without commercial bumblebees for some pathogens, but the results were only consistent between early and late season for Apicystis bombi and Black queen cell virus. Apicystis bombi had a higher prevalence (2,0 and 3,0 for late and early season) in areas with commercial bumblebees. Black queen cell virus had a higher prevalence (1,3 and 1,6 for late and early season) in areas without commercial bumblebees. For the seasonal fluctuation a significant difference between early and late season could be seen for the pathogens – Crithidia bombi, Apicystis bombi, Locustacarus buchneri and Sphaerularia bombi.

A prototype of a haptic and virtual reality simulator has been developed for simulation of the bone milling and material removal process occurring in several operations, e.g. temporal bone surgery or dental milling. The milling phase of an operation is difficult, safety critical and very time consuming. Reduction of operation time by only a few percent would in the long run save society large expenses. In order to reduce operation time and to provide surgeons with an invaluable practicing environment, this licentiate thesis discusses the introduction of a simulator system to be used in both surgeon curriculum and in close connection to the actual operations. The virtual reality and haptic feedback topics still constitute a young and unexplored area. It has only been active for about 10-15 years for medical applications. High risk training on real patients and the change from open surgery to endoscopic procedures have enforced the introduction of haptic and virtual reality simulators for training of surgeons. Increased computer power and the similarity to the successful aviation simulators also motivate to start using simulators for training of surgical skills. The research focus has been twofold: 1) To develop a well working VR-system for realistic graphical representation of the skull itself including the changes resulting from milling, and 2) to find an efficient algorithm for haptic feedback to mimic the milling procedure using the volumetric Computer Tomography (CT) data of the skull. The developed haptic algorithm has been verified and tested in the simulator. The visualization of the milling process is rendered at a graphical frame rate of 30 Hz and the haptic rendering loop is updated at 1000 Hz. Test results show that the real-time demands are fulfilled. The visual and haptic implementations have been the two major steps to reach the over all goal with this research project. A survey study is also included where the use of VR and haptic simulators in the surgical curriculum is investigated. The study starts with a historical perspective of the VR and haptic topics and is built up by answering different questions related to this topic and the implementation of simulators at the medical centres. The questions are of general concern for those developing surgical VR and haptic simulators. Suggested future work includes modelling, development and validation of the haptic forces occurring in the milling process and, based on this, implementation in the simulator system. Also, further development of the simulator should be done in close cooperation with surgeons in order to get appropriate feedback for further improvements of the functionality and performance of the simulator. / QC 20101112

Background: Urinary tract infection (UTI) is a common bacterial infection that is usually diagnosed by symptoms such as dysuria and frequency, and the golden standard is to take a urine culture to identify bacteria that may cause UTI. This method was founded with the idea that normal urine is sterile, but this is now being questioned because of growing evidence of a urinary microbiota thus giving a new approach to methods for UTI diagnosis. Aim: To identify and re-evaluate findings of bacteria from urine cultures in the ongoing paradigm shift of a potential urinary microbiome, and dysbiosis as a cause for UTI. Materials and Methods: This study used MALDI-TOF MS to identify approximately 250 bacteria isolates that had been cultured by Expanded Quantitative Urine Culture (EQUC) from 162 women with symptoms of cystitis. EQUC had allowed the bacteria to grow in both CO2 and anaerobic conditions, which differs from standard techniques.   Results and Conclusion: Escherichia coli and Enterococcus faecalis dominated the results of most frequently identified bacteria. However, other bacteria were commonly present within the same culture which is traditionally considered as contamination but may now indicate a urinary flora. Anaerobic bacteria – such as Porphyromonas sp. – were also identified, but their connection to UTI is unclear. Lactobacillus sp. – which are associated with a healthy flora in women – were found in urine cultures and often in smaller quantities which could suggest dysbiosis. More research on Lactobacillus sp. and their correlation with UTI is suggested for a more accurate indication of urinary dysbiosis in women.

Treponema pallidum (T. pallidum) is the bacteria that causes syphilis, which is a sexually transmitted disease that might give the carrier a multisystemic infection. A combination of serological tests is needed to put a diagnosis of T. pallidum infection, because the bacteria cannot be cultured in vitro. Serological tests categorize intro nontreponemal test and treponemal test, which both are needed for a correct diagnosis. The principle of a nontreponemal test is the measure of antibodies directed towards lipoid antigens, whereas the principle of a treponemal test is the measure of antibodies directed towards specific T. pallidum antigens. One kind of a treponemal test that are used are treponemal antibody test which is a manual agglutination test. At Sundsvall County hospital TP-PA was used for detection of antibodies, this kit is no longer available for purchase which means that a new kit must be validated to take its place. The purpose of this study was to evaluate the possible replacing kit, TP-HA. To validate TP-HA, ten positive serum samples were analyzed and compared with the two kits. A positive kit-control were also used to confirm the results of this study. The results showed that TP-PA was more sensitive than TP-HA according to a Wilxocon ranking test (p<0.05). Because TP-PA has to be replaced due to stricter in vitro diagnostic regulation (IVD-R laws), TP-HA will be taking its place even though it differs in sensitivity. This might effect tests with a lower titre, they might give a negative result even though the test has a low concentration of antibodies which needs to be further evaluated.

Background: Cystic fibrosis (CF) is a hereditary disorder caused by genetic mutations, which affect the chloride ion channels, leading to disrupted salt balance in different organs. A lack of properly functioning chloride ion channels can lead to formation of thick mucus, which hinders organ function, especially in the lungs where repeated inflammation occurs. Early diagnosis is critical to prevent further deterioration of the patient's condition. Current method of analysis of CF diagnostics uses conductivity meters to measure sweat electrolytes. However, current guidelines suggest using a chloridometer to directly measure chloride concentration, is the most reliable marker of cystic fibrosis. The aim of this project was to conduct a comprehensive evaluation of the new instrument's safety, reliability, validity, and conformity of the reference range to international chloride meter guidelines. Additional aims were to investigate the effect of storage conditions on sweat chloride concentration levels and examine the effect of increased salt intake on sweat test results. The study recruited healthy participants and took samples of their sweat by inducing sweat gland secretion. The chloride ion concentration was determined using a coulometric method. The results of the study found that the new method was reliable and matched international protocols. It also revealed that an increased salt consumption can impact chloride concentration in sweat, but not to an extent that it can affect medical decisions. Additionally, the study demonstrated that sweat samples can be frozen for up to two weeks without affecting the outcome of the chloride determination.

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Sleep is very important for health and academic performance. The amount of sleep is variable and depends among other things, on the age of the individual. Today there are many varieties of motion sensors that can be used in healthcare and clinical research for sleep registration, such as Actigraph GT9X Link and Actiwatch 2. The purpose of this study was to compare these two sensors, Actigraph and Actiwatch, for sleep registration by analyzing different sleep variables. The more precise sensor of the two will be used in the Energymetabolic laboratory in the case of survey or research. The method involved participants wearing both sensors for two nights and one day and trying to live as normal as possible. A sleep diary was used to complete the registrations, with related questions about sleep. There were 17 healthy participants between the ages 21 and 31. The results showed that the registrations from both sensors have varying spread and deviations. Overall the results were similar between the sensors and the sleep diary, and it was shown that there is no significant difference between the sensors. However, Actigraph deviated on most occasions from the diary. The Energymetabolic laboratory have access to many Actigraph GT9X Link sensors. This means that an evaluation has to be made to decide whether the cost of replacing the Actigraph GT9X Link sensors with Actiwatch 2 is worth it for the small improvement in precision.

Neuroendocrine tumours generally show an overexpression of somatostatin receptors on their cell membranes, mainly subtype 2. This is taken advantage of in diagnosis and therapy by using synthetic somatostatin analogues that can be labelled with radionuclides to visualize and treat tumours with an overexpression of somatostatin receptors. The method traditionally used for visualization is somatostatin receptor scintigraphy (SRS) with 111In-DTPA-octreotide but this method is gradually being substituted by 68Ga-DOTATOC PET/CT. To evaluate patients for peptide receptor radionuclide therapy, it is mandatory for the patient to be examined by both methods. In the evaluation, the tumours are graded according to the Krenning scale on the images from the SRS. Patients with sufficient tumour uptake of somatostatin analogues are eligible for peptide receptor radionuclide therapy (PRRT). The aim of this study was to compare the tumour’s Krenning scores from SRS to the Krenning scores, quantitative indices and TNR-values from the 68Ga-DOTATOC PET/CT images. This was done to investigate if the Krenning scale could be applied to PET/CT enabling the patient to undergo only PET/CT for diagnosis and evaluation prior to PRRT. This study, including 28 patients, found no strong correlation between the Krenning scores from the SRS and the scores from 68Ga-DOTATOC PET/CT. However, a better correlation was shown between the Krenning scores from SRS and TNR-values where the quantitative indices SUVmax and SUVmean were divided with the SUVmean of the spleen. These findings could be worth exploring further in future studies, incorporating larger number of patients.

Background: Valvular heart disease (VHS) can be lethal. An auscultated murmur could be a first indication of VHS. Lately auscultation has been evaluated as non-accurate, while a murmur also can be normal/functional. The next step of verifying VHS, is a transthoracic echocardiography (TTE). The echocardiography clinic at Uppsala University Hospital has seen a lot of non-pathological referrals regarding murmur evaluation. Therefore, a fast-track screening TTE, performed by a biomedical scientist was of interest. Aim: The aim was to evaluate pathological possibility, regarding remitted patients with a new heart murmur. Material: The clinical quality work was based on remitted patients of ages 18 to 50. Sampling took place between November 2022 and Mars 2024, by Radiology Information System. Method: Type of murmur, outcomes and referring clinic was documented. Normal outcome group consisted of: absent VHS and mild VHS. Pathological outcome group consisted of: moderate and severe VHS. Probability was calculated based on systolic- and non-specified murmur. Result: Normal outcome group had 116 referrals and pathological outcome group had three referrals. Possibility of a pathological outcome became 2,5 %. Majority of the referrals came from the primary care (92 %). Conclusion: A systolic- and non-specified murmur had low possibility of a pathological outcome, which could indicate that a shorter screening TTE by a biomedical scientist is an option. A limitation was that the type of the remitted murmur could not be trusted. Majority of the referrals came from the primary care, which indicates that further clinical work at these facilities is necessary.

Biocides and antibiotics are commonly used in Irish agriculture. This could lead to accumulation at sublethal levels in water and resistance development. The risk of this has earlier been assessed as non-existent. However, resistant strains have been found in Irish farm waste water. Due to possible horizontal gene transfer between bacterial populations the risk may be higher. Antibiotic resistance mechanisms have worked against certain biocides and antibiotic resistant strains have in certain cases showed reduced biocidal susceptibility. The aim of this project was to characterise bacterial populations from Irish aquatic samples and investigate their susceptibility to agriculturally common biocides and relevant antibiotics. Isolates from Dublin Bay water samples (n=15) were characterised using basic techniques. Minimum inhibitory concentration (MIC) tests with a broth microdilution method were performed with eight biocides and complimentary minimum bactericidal concentration (MBC) tests. Antibiotic disc diffusions were performed with eight antibiotics. The samples contained gram-negative isolates (n=3), Staphylococcus aureus (n=1) and Bacillus spp. (n=8) isolates. All isolates were on average resistant towards methylated spirits and iodine at the 2% v/v starting concentration. MIC values for Tri Scrub and the generic biocide were high. All MBC values were on average higher than the corresponding MIC values. A significant amount of the Bacillus spp. isolates were resistant towards β-lactams. As there is statistical uncertainty around the results, further investigations are needed. In conclusion, a trend of both high MIC and MBC values while showing resistance towards the largest number of antibiotics could be seen in Bacillus spp. isolates.