The aim of this study was to develop improved methods to detect viable Coxiella burnetii in wastes from livestock production. The impetus for this work arose because there is a significant risk of infection for humans attributed to contact with waste products from the livestock production industry. This situation is further compounded by the lack of suitable tools to detect viable C. burnetii in these wastes. In addition, effective disinfection strategies for livestock wastes are also required to reduce the risk of infection with C. burnetii for individuals that come into contact with these waste products. A quantitative real-time PCR system (qPCR) with high sensitivity and specificity was developed to detect the C. burnetii in environmental samples associated with domestic ruminants and native Australian marsupials. Different detection chemistries and procedures were evaluated based on their sensitivity, specificity and reproducibility. Overall it was found that the TaqMan PCR targeting the IS1111a locus provided the most sensitive and reproducible test. The Geneworks PowerSoil(tm) DNA isolation kit provided the best compromise between reproducibility and recovery of DNA from livestock wastes. When combined, the IS1111a TaqMan qPCR and Geneworks PowerSoil DNA Extraction Kit provided a test which was capable of detecting as few as two C. burnetii genome equivalents in 0.2g of soil or faeces. Coxiella burnetii has been shown to display extreme resistance to environmental exposure. Therefore, assessment of the viability of the organism in environmental matrices is more useful for risk assessment programs than detection of DNA alone. A quantitative reverse transcriptase PCR was developed that was able to detect viable C. burnetii cells in soil. The sensitivity of the assay was enhanced by heat-treating the soil samples prior to extraction of RNA. The factor most often associated with transfer of C. burnetii to humans is exposure to livestock or their waste. Therefore, decontamination of waste from livestock production industries is a key factor in preventing outbreaks of Q fever. A system was developed to determine the efficacy of various disinfectant treatments against the environmental pathogen C. burnetii. Treatments evaluated included sodium hypochlorite, ozone, ultraviolet light, peracetic acid (PAA), and Virkon S®. Sodium hypochlorite at a concentration of 0.1 mM reduced the infectivity of C. burnetii by over 92% while treatment with the same sodium hypochlorite concentration in wastewater showed significantly reduced efficacy. Despite this reduced potency, sodium hypochlorite is still useful for control of C. burnetii in the liquid waste of animal production. Commercially available ELISA and CFT assays exist for ruminants but there are no immunological tests available for detecting C. burnetii in marsupials even though Australian marsupials are known to be susceptible to C. burnetii. An indirect ELISA for detecting anti-Coxiella antibodies in kangaroos was developed. Paired serum and faecal samples were taken from 379 ruminants from Western Australia and the serum was tested with a commercially available ELISA and the complement fixation test while the faeces was tested using the qPCR developed during this study. Paired serum and faecal samples were taken from 343 kangaroos from WA and were tested with the antibody-ELISA developed during this study and by qPCR. A very low prevalence of anti-Coxiella antibodies was observed in the ruminants sampled and results from immunological tests correlated poorly with qPCR data. The development of an ELISA for use with kangaroo serum was problematic because of the lack of reference sera from animals known to be infected with C. burnetii. Despite this results from the ELISA developed suggested that the apparent seroprevalence in the WA animals surveyed was approximately 34%. Results from testing kangaroo faeces with the qPCR correlated poorly with the results from the antibody-ELISA. These data suggest that kangaroos may be a significant reservoir of C. burnetii in Western Australia and due to cohabitation of kangaroos and domestic ruminants, may provide a link between the wildlife and domestic cycles of C. burnetii.

Until recently, occurrence of milk fever (MF) has been attributed to prepartum calcium intake. However, researchers in Iowa have concluded that high prepartum dietary potassium (K) is the major cause of MF. Potassium concentrations have been increasing on manure fertilized soils over the last 20 years. Grasses grown on these fields mirror the increase in available soil K. When high K forages are fed to dry cows, it has been associated with increased MF in some, but not all cases. Our objective was to identify factors that differed between low and high occurrence of disorders when high K forages were fed. For 1 yr, monthly interviews were conducted on 10 dairies in Western Oregon. Close-up rations, dystocia rating, crowding, and cow comfort data were collected. Cows were diagnosed healthy or having one or more of the following metabolic diseases: MF, retained fetal membranes (RFM), and left displacement of the abomasum (LDA). Feedstuffs were collected each month and analyzed for dry matter, crude protein (CP), acid detergent fiber (ADF), and macrominerals. Of the multiparous cows (n=3,587) included, incidences of MF, RFM and LDA were 3.6, 11.3, and 1.5%, respectively. Increasing MF occurrence was associated with uncomfortable conditions, dystocia, increasing prepartum dietary Na and ADF, and increasing Ca to P ratios; there was also a dietary K by Mg concentration interaction. Increased dietary concentrations of Mg can prevent MF if dietary K is <2.6%. In addition, dietary conditions of K >2.6% and Mg >0.4% increase the occurrence of MF. Inversely, high concentrations of K can prevent MF in a Mg deficient diet. Dietary factors associated with an increase in the occurrence of RFM are increased dietary concentrations of Ca, S, CP, use of anionic salts, and the K forage source. Factors associated with an increase in hypocalcemia in the postpartum cow are crowding, uncomfortable housing, and a dietary K by Mg concentration interaction. In conclusion, improved cow comfort, and the addition of Mg to high K forage diets could decrease the risk of the cow exhibiting MF, RFM, and hypocalcemia. / Graduation date: 1999

As early as in 1915, 1931 and 1942, there were serious outbreaks of dengue fever occurred to the entire island of Taiwan. In 1942, there were five million people in the whole Taiwan infected with dengue fever. After the significant outbreaks of dengue fever in the entire Kaohsiung District of Taiwan Island in 1987 and 1988, except that the regional epidemics occurred in three places of Northern and Central Taiwan, namely Jhonghe City of Taipei County, Taichung City and Taipei City, all other regional outbreaks of dengue fever mostly occurred in Kaohsiung County or City, Tainan City and Pingtung County. Apparently, the epidemic situation of dengue fever has become an epidemic disease of Southern Taiwan, and gradually developed as a ¡§community disease¡¨ or ¡§environmental disease.¡¨ Coincidently, the year of 2006 was a peak period for the propagation of dengue fever. Under the circumstances that the number of dengue fever patients was getting higher and higher, both the government and the competent authorities took compulsory measures or epidemic prevention acts (such administrative acts as insecticide spraying against mosquitoes in residences after forceful unlocking of doors, emergency public spraying of chemicals against mosquitoes, taking blood tests, and so on). It seemed so obvious that the liberty of living, liberty of property and personal liberty, which are the basic rights of civilians under the protection of the Constitution, were involved. Besides, whenever dengue fever is epidemic, it is common to see conflicts (arguments) over the door unlocking and insecticide spraying problems frequently happened between the citizens and the government departments concerned. In the legal aspect, it is necessary to review whether these acts comply with the provisions of the Constitution. Apart from collecting the related local and foreign literatures to investigate the change of the administrative system of epidemic control, the study introduces the details of the three major directions in taking epidemic prevention measures against dengue fever in Taiwan, including such administrative and management acts as the preventive measures to be taken during the ordinary days, the epidemic control measures after receiving the report of dengue fever cases, and the epidemic prevention measures after the confirmation of dengue fever cases. In addition, regarding the various compulsory epidemic control measures taken by the health administrative authorities, such as the problems of forceful unlocking of doors for intrusion of residences, forceful (emergency) spraying of insecticide, and the compensation for the loss of property rights after the death of real estate owners¡¦ animals or plants caused by the chemical spraying, etc., the study reviews the reference of laws and the proper legal procedures to be complied with. It includes the theories and principles of the Constitution that Taiwan government should comply with. Although the paper encounters limitations in the studying process, it is still hoped that from the limited literatures with limited information, major directions can be found out as a reference for the competent authorities or scholars to improve or solve the abovementioned problems in the later days. It is also expected that the later researchers can have further understanding of the implementation or practical aspects of the epidemic prevention measures of dengue fever, and finally achieve in-depth development and breakthrough in the studies of the solutions. Keywords: dengue fever, basic human rights, epidemic prevention measures

The impact of Community - on health is one of the core issues of sociology and social epidemiology. Dengue fever, whose occurrence and eradication highly depend in the environment of the community, provides an opportunity to explore the relationship between the community and the health. This study focuses on the outbreak of Dengue fever in 2006 in Kaohsiung City and analyzes how community factors affect the cluster infections of. This study uses the theories of social capital and social vulnerability to explore the effect population, socioeconomic status, and public participation in the community on cluster infection. Social capital is formed by community members through participating in public affairs and voluntary organizations; social vulnerability refers to the overall socio-economic conditions that shape the ability of community to respond to the undesirable events. The social capital index was constructed composed of the Community Development Association, the voting rate of warden elections. Social vulnerability index is based on the compositions of the population, including the proportion of elderly males, population change rate, ratio of low-income households the proportion of dependents than average household income. In addition to consideration the spatial characteristics of dengue fever, this thesis uses Moran's I and LISA test dengue fever outbreaks, and correct the result by Geographically Weighted Regression. The results shows the impact of community factors on the health of residents, provide health sociology and control of dengue fever different reflections direction.

Coxiella burnetii is an intracellular pathogen that can cause both acute and chronic disease (Q fever) in humans and infects many animals with varying clinical illness and persistence. A guinea pig aerosol-challenge model of acute Q fever was developed using infection with C. burnetii across a 5-log range of challenge doses. Clinical signs included fever, weight loss, respiratory difficulty, and death, with degree and duration of response corresponding to dose of organism delivered. Histopathologic evaluation revealed coalescing panleukocytic bronchointerstitial pneumonia 7 days after a high-dose challenge, resolving to multifocal lymphohistiocytic interstitial pneumonia by 28 days. Clinical and pathologic changes noted in these guinea pigs were comparable to those seen in human acute Q fever, making this an accurate and valuable animal model. This model was used to compare the relative virulence of eight isolates from four different genotypic groups: I (RSA493, RSA334, and RSA270), IV (Q177 and Q173), V (Q212 and Q217), and VI (5J108-111). Guinea pigs infected with group I acute-diseaseassociated isolates had severe respiratory disease, while no to moderate clinical illness was observed in animals given group IV or V chronic-disease-associated isolates. 5J108- 111 appeared avirulent. These data suggest that C. burnetii isolates have a range of disease potentials and support a distinction in strain virulence between established genotypic groups, though isolates within the same genomic group cause similar pathologic responses. Heterologous protection was confirmed by cross vaccination and challenge with RSA493 and Q217. A marked non-specific suppression of lymphoproliferation was noted at 14 and 28 days post infection with RSA493; similar suppression was seen after infection with Q173 and Q212 but not 5J108-111. Proinflammatory cytokines IFN-γ and TNF-α were produced during early C. burnetii infection, at which time anti-inflammatory cytokines TGF-β and IL-10 were repressed. A vaccine made from phase I C. burnetii was found to be completely protective against lethal infection in the guinea pig model, while vaccination with killed phase II organisms conferred only partial protection, preventing death and reducing but not precluding fever and respiratory illness. Protective vaccination significantly stimulated cell-mediated immunity and elicited increases in IFN-γ, TNF-α, and IL-12p40 mRNA levels.

published_or_final_version / Microbiology / Master / Master of Philosophy

Spatio-temporal systems with heterogeneity in their structure and behavior have two major problems. The first one is that such systems extend over very large spatial and temporal domains and consume a lot of resources to simulate that they are infeasible to study with current platforms. The second one is that the data available for understanding such systems is limited. This also makes it difficult to get the data for validation of their constituent processes while simultaneously considering their global behavior. For example, the valley fever fungus considered in this dissertation is spread over a large spatial grid in the arid Southwest and typically needs to be simulated over several decades of time to obtain useful information. It is also hard to get the temperature and moisture data at every grid point of the spatial domain over the region of study. In order to address the first problem, we develop a method based on the discrete event system specification which exploits the heterogeneity in the activity of the spatio-temporal system and which has been shown to be effective in solving relatively simple partial differential equation systems. The benefit of addressing the first problem is that it now makes it feasible to address the second problem.We address the second problem by making use of a multilevel methodology based on modeling and simulation and systems theory. This methodology helps us in the construction of models with different resolutions (base and lumped models). This allows us to refine an initially constructed lumped model with detailed physics-based process models and assess whether they improve on the original lumped models. For that assessment, we use the concept of experimental frame to delimit where the improvement is needed. This allows us to work with the available data, improve the component models in their own experimental frame and then move them to the overall frame. In this dissertation, we develop a multilevel methodology and apply it to a valley fever model. Moreover, we study the model's behavior in a particular experimental frame of interest, namely the formation of new sporing sites.

No description available.

No description available.

The spirochete Borrelia hermsii avoids the immune response of its mammalian host through multiphasic antigenic variation. Serotype specificity is determined by Variable major proteins (Vmp), in the outer membrane. Through a non reciprocal recombination between linear plasmids, a formerly silent vmp gene replaces another vmp gene at a telomeric expression locus downstream from a common expression site. B. hermsii before and after the switch from serotype 7 to serotype 21, was examined in detail. The nucleotide sequence of the vmp7 and vmp21 genes and flanking regions was determined. The vmp7 and vmp21 are 77% identical in their coding sequence, and the deduced translation products are 63% identical. No antigenic cross reactivity is observed between Vmp7 and Vmp21. This suggests a folding of the proteins in which the similar regions are buried, and not exposed when it is presented at the bacterial surface. Vmp7 and Vmp21 have consensus sequences of prokaryotic lipoproteins and are processed as such when expressed in E. coli. The 5' regions of silent and expressed vmp7 and vmp21 were compared. Silent and active vmp7 and vmp21 genes shared a block of homologous sequence at their 5' ends. Sequences upstream of silent vmp7 and vmp21 genes lacked a promoter and differed substantially from each other. In this antigenic switch a vmp gene was activated by a recombination event which placed it downstream of a promoter. The vmp gene promoter is preceded by a poly(dT dA) ran and three imperfectlyrepeated elements of 2 kb. Each of the 2 kb repeats contains inverted repeats of approximately 0.2 kb at their termini. There is no evidence of the presence of similar elements elsewhere in the genome of B. hermsii. One or more of these elements may stimulate vmp gene switch or expression. The African relapsing fever species Borrelia crocidurae and the American species B. hermsii display many similarities. In both species the vmp genes are localised to linear plasmids, and the vmp genes are activated on the transcriptional level. The nucleotide sequence of their expression sites, however, are not related. Still, the possibility that the switch is mechanistically similar in B. crocidurae and B. hermsii, cannot be ruled out. The binding of B. crocidurae causes aggregation of erythrocytes around the spirochete. The aggregation is reminiscent of the erythrocyte rosetting seen in malarial infections. The erythrocytes at the B. crocidurae surface may protect them from clearance by the host. Thus, the rosetting may constitute an additional mechanism in B. crocidurae for the evasion of the immune reaction. / <p>Diss. (sammanfattning) Umeå : Umeå universitet, 1994, härtill 5 uppsatser.</p> / digitalisering@umu