Submitted in partial compliance with the requirements for a Master Degree in Technology: Chiropractic at the Durban University of Technology, 2009. / Background: Research has indicated the majority of bacteria on chiropractic adjustment beds (beds), can persist on dry inanimate surfaces for months. Thus, insufficient disinfection procedures create continuous sources of pathogens endangering patients and healthcare workers alike. This research study aimed to assess the beds as reservoirs for micro-organisms, at a chiropractic teaching clinic (clinic) in South Africa. Method: A selection of samples obtained from the headrests and armrests of the beds were serially diluted, plated in duplicate (using the spread plate technique) and incubated for 24-48 hours at 37°C. After inspection for the presence of micro-organisms, those present were enumerated to determine their quantities, the microbial build-up throughout the day, as well as the degree of the transmission from the patients to the beds during treatment. The incidence of the micro-organisms was established, along with their identities, using microscopic and macroscopic characteristics. These micro-organisms were also used to assess the efficacy of the disinfectant currently in use by the clinic. Results: Microbial growth was present on 89.4% of the beds sampled. The quantities of the micro-organisms increased significantly (p=0,027) from 7:30 am to 16:30 pm, with the median increasing from 25 colony forming units (cfu) / cm2 to 714 792 cfu/ cm2. The microbial build-up was highly significant (p<0.001), with a median of 346 cfu/ cm2 at 7:30 am and 10:30 am; increasing to 162 291 cfu/ cm2 by 13:30 pm and 250 million cfu/ cm2 by 16:30 pm. There was also a significant increase (p<0.001) in the quantity of micro-organisms during treatment with a median of 0 cfu/ cm2 before treatment that rose to 23 479 cfu/cm2 after treatment, indicating that the micro-organisms present on the beds were being deposited by the patient`s skin during the treatment. The most prevalent micro-organisms identified were Staphylococci and Serratia, with an average of 59% and 40% of colonies; while Micrococci and Bacilli were relatively uncommon. No growth was evident after 5 minutes of exposure to the disinfectant during the growth inhibition test. For the Kirby Bauer test, the average size of the zone of inhibition increased as the dilution decreased. The disinfectant is effective but more so against the Gram-positive than the Gram-negative bacteria. The disinfectant was 5,0, 5,5 and 5,6 times more effective than phenol in eradicating Staphylococci, Serratia and Bacilli, respectively. Conclusions and Recommendations: This study showed that micro-organisms were present on the beds. Staphylococci and Serratia have been implicated in many healthcare associated infections. The present disinfectant is effective, but should be used in between every patient. A different or additional disinfectant that is more effective against the Gram-negative bacteria should be considered for future use.

Salmonella enteritidis (S. enteritidis) is a bacterial pathogen of chickens, and is currently one of the leading causes of human food poisoning in the world. It is believed that contaminated poultry products, especially eggs and egg products, have been responsible for the dramatic increase in the incidence of this Salmonella serotype. Detection of S. entertidis has conventionally involved bacteriological examination of samples, yet these procedures are time-consuming which could lead to the rapid spread of S. enteritidis through commercial flocks and potentially cause a human health risk. A number of alternative detection techniques, mostly based on serological methods, have been reported as effective diagnostic assays. However, some of these reports have not been supported by representations of SDS-PAGE gels or Western blots. The objective of this study was the evaluation of these serological techniques as well as a PCR amplification technique, which has been reported to show promising results as a diagnostic method. The techniques discussed in these reports were evaluated with regards to how rapid they were, their specificity and their potential for use in local diagnostic laboratories. Antigens from the outer surface of S. enteritidis were purified by several methods and their antigenicity was tested by separating the antigens by means of SDS-PAGE, followed by Western blotting using sera of chickens infected with S. enteritidis. A high degree of cross reactivity was observed with many of the antigens tested, especially the lipopolysaccharides (LPSs) and outer membrane proteins (OMPs) which had previously been reported as containing antigens which could be used for specific detection of S. enteritidis. This cross-reactivity could be explained by the conserved nature of many of the LPS and OMP antigens among the Salmonella serotypes tested. A fimbrial antigen, SEF14, which has been reported as a novel antigen, was seen as a prominent band at 14.3 kDa and was found to react with antibodies against S. enteritidis, yet not to the specificity levels described in previous reports. PCR amplification of the sefA gene sequence, which encodes for the SEF14 fimbrial antigen, was found to give a predicted product of 310 bp when using a previously described oligonucleotide primer pair. This amplified product was found to be specific for S. enteritidis and other serogroup D Salmonella serotypes that are not poultry pathogens The cross-reactivity observed with many of the serological techniques used in this study, meant that detection of S. enteritidis infection in chickens was considerably hindered. However, the identification of further novel antigens by serological means, could result in the development of new vaccines. The specificity and speed afforded by PCR amplification indicated that this technique showed excellent potential for use in local diagnostic laboratories. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.

This research validates electro-disinfection as a potential secondary ballast water treatment technology. Electricity applied to bacteria laden water produced bactericidal effects, reactive oxygen species and chlorine generation which annihilated bacteria. Evaluation of electro-disinfection experiments showed titanium electrodes had the maximum kill efficacy while disinfection with aluminum and stainless steel electrodes had lesser kill efficacy. A continuous flow electro-disinfection reactor was evaluated utilizing artificial brackish and fresh ballast water. Brackish water had a 100% bacteria kill efficiency utilizing titanium electrodes at a current density of 10 mA/cm2. Fresh water was augmented with the addition of salt to increase its electrical conductivity from 232 μS/cm to 873 μS/cm to ascertain 100% bacteria kill efficiency with titanium electrodes and a current density of 9.8 mA/cm2.

A leptospirose é uma zoonose causada por bactérias patogênicas do gênero Leptospira. Apesar de sua importância, a patogênese e virulência permanecem não elucidadas. As leptospiras não apresentam proteases conhecidas de degradação de matriz extracelular, atividade crucial para a penetração e disseminação nos hospedeiros. Assim, foi proposta a investigação da interação de leptospiras com plasminogênio/plasmina e as implicações para a infecção. As leptospiras capturam plasminogênio na superfície, e este é convertido à plasmina por ativadores do hospedeiro. A plasmina associada propicia degradação de componentes de matriz extracelular, habilidade de penetração e evasão imune. Adicionalmente, as leptospiras estimulam a expressão de ativadores de plasminogênio e metaloproteases de matriz. Os resultados contribuem para o conhecimento do processo infeccioso das leptospiras, descrevendo um novo mecanismo de patogenicidade. / Leptospirosis is a zoonosis caused by pathogenic bacteria from genus Leptospira. Despite its importance, the pathogenicity and virulence remain to be elucidated. The leptospires do not present known proteases able to degrade extracellular matrix, an activity essential for the penetration and dissemination within the hosts. Therefore, we proposed the investigation of the leptospiral interaction with plasminogen/plasmin and its implications for infection. Leptospires capture plasminogen on the surface, which is converted to plasmin by hosts activators. Surface-bound plasmin confers extracellular matrix components degradation, penetration ability and immune evasion. Additionally, leptospires stimulate plasminogen activators and matrix metaloproteases expressions. The results constitute one possible mechanism that contributes to the invasion process and the rapid dissemination of Leptospira.

Abstract not available

Bacteria cause significant morbidity and mortality throughout the world, including deadly diseases such as tuberculosis, meningitis, cholera, and pneumonia. Timely and accurate bacterial identification is critical in areas such as clinical diagnostics, environmental monitoring, food safety, water and air quality assessment, and identification of biological threat agents. At present, there is an established need for high throughput, sensitive, selective, and rapid methods for the detection of pathogenic bacteria, as existing methods, while nominally effective, have failed to sufficiently reduce the massive impact of bacterial contamination and infection. The work presented in this thesis focuses on addressing this need and augmenting conventional microorganism research through development of mass spectrometry (MS)-based proteomic applications. MS, a well established tool for addressing biological problems, offers a broad range of laboratory procedures that can be used for taxonomic classification and identification of microorganisms. These methods provide a powerful complement to many of the widely used molecular biology approaches and play critical functions in various fields of science. While implementation of modern biomolecule-identifying instrumentation, such as MS, has long been postulated to have a role in the microbiology laboratory, it has yet to be accepted on a large scale. Described in this document are MS methods that erect strong foundations on which new bacterial diagnostics may be based. A general introduction on key aspects of this work is presented in Chapter 1, where different approaches for detection of pathogenic bacteria are reviewed, and an overview regarding MS and microbial identification is provided. Chapter 2 presents the first implementation of microbial identification via rapid, open air Direct Analysis in Real Time MS (DART MS) to generate ions directly from microbial samples, including the disease-causing bacteria, Coxiella burnetii, Streptococcus pyogenes, and Escherichia coli. Chapter 3 expands on whole cell C. burnetii MS analysis and presents a rapid differentiation method to the strain-level for C. burnetii using mass profiling/fingerprinting matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS and multivariate pattern recognition. Chapter 4 presents a unique "top-down" proteomics approach using 15N-labeled bacteriophage amplification coupled with MALDI-TOF MS as a detector for the rapid and selective identification of Staphylococcus aureus. Chapter 5 extends the idea of using isotopically labeled bacteriophage amplification by implementing a "bottom-up" proteomics approach that not only identifies S. aureus in a sample, but also quantifies the bacterial concentration in the sample using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS) as a detector. In conclusion, Chapter 6, summarizes and contextualizes the work presented in this dissertation, and outlines how future research can build upon the experimentation detailed in this document.

Mannheimia haemolytica is an opportunistic pathogen in cattle and the main bacterial agent in bovine respiratory disease. Despite its economic importance, few studies have characterized the genetic diversity of M. haemolytica, particularly from feedlots. Three genotyping techniques (BOX-PCR, (GTG)5-PCR and PFGE) were compared to discriminate M. haemolytica and strains from the family Pasteurellaceae. PFGE was the most discriminating and repeatable, although BOX-PCR was most accurate in clustering isolates together according to species. Mannheimia haemolytica was isolated from nasal swab samples collected from cattle upon entry and exit from two feedlots in southern Alberta. These were characterized by PFGE and antimicrobial susceptibility using a disk-diffusion assay. Select gene determinants were screened for using PCR. PFGE analysis revealed the isolates to be highly diverse. Ten percent of the isolates exhibited resistance. At present, the development and spread of antimicrobial resistance in M. haemolytica observed within the feedlots examined appears to be low. / xi, 116 leaves : ill. ; 29 cm

No description available.

Submitted in partial compliance with the requirements for a Master Degree in Technology: Chiropractic at the Durban University of Technology, 2009. / Background: Research has indicated the majority of bacteria on chiropractic adjustment beds (beds), can persist on dry inanimate surfaces for months. Thus, insufficient disinfection procedures create continuous sources of pathogens endangering patients and healthcare workers alike. This research study aimed to assess the beds as reservoirs for micro-organisms, at a chiropractic teaching clinic (clinic) in South Africa. Method: A selection of samples obtained from the headrests and armrests of the beds were serially diluted, plated in duplicate (using the spread plate technique) and incubated for 24-48 hours at 37°C. After inspection for the presence of micro-organisms, those present were enumerated to determine their quantities, the microbial build-up throughout the day, as well as the degree of the transmission from the patients to the beds during treatment. The incidence of the micro-organisms was established, along with their identities, using microscopic and macroscopic characteristics. These micro-organisms were also used to assess the efficacy of the disinfectant currently in use by the clinic. Results: Microbial growth was present on 89.4% of the beds sampled. The quantities of the micro-organisms increased significantly (p=0,027) from 7:30 am to 16:30 pm, with the median increasing from 25 colony forming units (cfu) / cm2 to 714 792 cfu/ cm2. The microbial build-up was highly significant (p<0.001), with a median of 346 cfu/ cm2 at 7:30 am and 10:30 am; increasing to 162 291 cfu/ cm2 by 13:30 pm and 250 million cfu/ cm2 by 16:30 pm. There was also a significant increase (p<0.001) in the quantity of micro-organisms during treatment with a median of 0 cfu/ cm2 before treatment that rose to 23 479 cfu/cm2 after treatment, indicating that the micro-organisms present on the beds were being deposited by the patient`s skin during the treatment. The most prevalent micro-organisms identified were Staphylococci and Serratia, with an average of 59% and 40% of colonies; while Micrococci and Bacilli were relatively uncommon. No growth was evident after 5 minutes of exposure to the disinfectant during the growth inhibition test. For the Kirby Bauer test, the average size of the zone of inhibition increased as the dilution decreased. The disinfectant is effective but more so against the Gram-positive than the Gram-negative bacteria. The disinfectant was 5,0, 5,5 and 5,6 times more effective than phenol in eradicating Staphylococci, Serratia and Bacilli, respectively. Conclusions and Recommendations: This study showed that micro-organisms were present on the beds. Staphylococci and Serratia have been implicated in many healthcare associated infections. The present disinfectant is effective, but should be used in between every patient. A different or additional disinfectant that is more effective against the Gram-negative bacteria should be considered for future use.

HIV infection has risen to pandemic proportions. Interleukin-10 (IL-10), a potent antiinflammatory cytokine has been shown to enhance the establishment and persistence of chronic viral infections through inactivation of effector antiviral immune responses and it may also directly influence HIV-1 replication in cells of diverse lineages. IL-10 promoter polymorphisms have been shown to affect HIV-1 susceptibility and pathogenesis. However, the underlying mechanisms are poorly understood. We investigated the relationship between IL-10 promoter variants, plasma IL-10 levels, and markers of disease outcome in chronically HIV-1-infected individuals. To investigate the mechanistic role of IL-10 and its genetic variants on HIV pathogenesis, we studied markers of activation on B cells, CD4+ and CD8+ T cells, and assessed effects on CD4+ T cell proliferation with and without blockade of the IL- 10 pathway. We used Taqman genotyping assays to genotype three IL-10 promoter single nucleotide polymorphisms (SNPs) in our study cohort. Baseline plasma IL-10 levels were measured using Luminex technology for 112 individuals. Viral load, CD4+ T cell counts and cytotoxic T lymphocyte (CTL) immune responses were measured at baseline. The rate of CD4+ T cell decrease was calculated in 300 individuals with a median follow-up of 25 months. CD38, CD95, Ki67, IgG and PD-1, markers of activation or exhaustion were measured on B cells, and CD38, CD95, Ki67, HLA-DR and PD-1 were measured on CD4+ and CD8+ T cells in a subset of 63 individuals. CD4+ T cell proliferation was measured using Carboxyfluorescein succinimidyl ester (CFSE) assays, following IL-10 receptor blockade in a subset of 31 individuals. The IL-10 -1082G, -592A and -3575 variants were observed at frequencies of 0.3, 0.34 and 0.23 respectively, in our study cohort. Plasma IL-10 levels were significantly higher in the - 1082GG group than in the combined AA/AG group (p=0.0006). There was a significant association between the 592AA genotype and a greater breadth of CTL responses compared to the CC and CA (p= 0.002 and 0.004 respectively). The -592AA genotype associated significantly with an attenuated loss of CD4 cells (p= 0.0496), with -592AA having the least change in CD4 cells per year. The median expression of HLA-DR, a marker of T cell activation was significantly higher in the-1082AA group for CD8 cells (p= 0.047), and the - 592AA group for CD4 T cells (p= 0.01). The median expression of IgG on the surface of B cells was significantly higher in the -1082GG genotype and the -592CC genotype (p=0.0183 and 0.0659 respectively). Overall, IL-10 variants correlated with IL-10 expression and CD4 decline during chronic HIV-1 infection. IL-10 promoter variants may influence the rate of HIV-1 disease progression by regulating IL-10 levels, which in-turn, may affect the breadth of CTL responses. Furthermore, the increased expression of HLA-DR and PD-1 on CD8+ and CD4+ T cells, indicates that lower IL-10 levels are associated with increased immune activation and immune exhaustion. The increased expression of IgG on B cells, suggests that in a setting of lower IL-10, there is possibly a bias towards a Th2 immune response. These data suggest a significant role for IL-10 genetic variants and IL-10 in HIV pathogenesis. Further studies to determine whether and how the IL-10 pathway may be manipulated for therapeutic or vaccine strategies for HIV are warranted. / Thesis (Ph.D.)-University of KwaZulu-Natal, Durban, 2012.