M. Tech. / Yeast such as Candida albicans are the major cause of human diseases such as genital thrush and oral thrush. Some of the genital isolates of C. albicans that were studied by Shah et al. (1991 & 1995) were found to produce the medically important immunosuppressing mycotoxin gliotoxin, which has potential important medical consequences. The biosynthesis of this mycotoxin is regulated and expressed by the presence of the gliP and gliZ genes, which were identified on the putative gene cluster of A. fumigatus. Most Candidal infections are treated using a single or a combination of antifungal agents such as amphotericin B (AmB), fluconazole, flucytosine, voriconazole, caspofungin, itraconazole, posaconazole and ketoconazole. The mode of action for these antifungal agents differs in terms of what molecule or processes are inhibited. The details of each antifungal agent and its mode of action are discussed in chapter 6 (page 54). These antifungal agents are usually recommended for the treatment of candidosis and currently the most common Candida spp. have developed resistance to these antifungal agents. The identification of Candida isolates was done using 2 different types of identification methods i.e., the chromogenic medium CHROMagar Candida and the biochemical test kit API 10 Candida. The chromogenic medium was inoculated with the Candida spp. supplied and incubated for 3 days at 37oC. The API 10C test strips were loaded with the culture suspension and incubated for 24 hours at 37oC. For the screening of gliotoxin, 2 supplemented mediums were used to cultivate the isolates that is the yeast extract sucrose (YES) and Eagles minimal essential medium (EMEM) and the isolates were grown at 37oC for 72 days. The methods that were used to identify the gliotoxin were thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) to quantify the levels of gliotoxin.

Thesis (MSc (Microbiology))--University of Pretoria, 2008. / Summary in English. Includes bibliographical references.

Background: Respiratory diseases are common worldwide, which are caused by various respiratory viruses. As symptoms caused by these viruses are similar, laboratory diagnosis is essential to distinguish the virus. Conventionally, respiratory viruses are isolated by cell culture with a panel of cell lines. However, handling of several cell lines is labour intensive, and the turnaround time of conventional culture is long. In previous study, the use of human colon adeno-carcinoma (Caco-2) in conventional culture was investigated. The study has proven that Caco-2 is generally susceptible to the eight common respiratory viruses, i.e. Adenovirus, Influenza A and B, Respiratory Syncytial virus, Parainfluenza virus 1, 2,3 and 4. As turnaround time of conventional culture is long; therefore, in this study, rapid shell vial culture using Caco-2 cells were evaluated. Moreover, the application of Caco-2 shell vial culture on recovering human metapneumovirus (hMPV) was also investigated. Materials and methods: This study consisted of four stages. First, recovery of viruses by conventional culture and shell vial culture of Caco-2 were compared. Specimens were added to conventional culture and shell vial simultaneously. For conventional culture, formation of CPE was examined daily and IF staining was performed when CPE was indicated; meanwhile, shell vial culture were incubated for seven days and stained with IF to detect infected cells. In stage two, the effect of incubating shell vial culture in rolling drum was investigated. Shell vials inoculated with the same specimen in duplicate were incubated in rolling drum and without rolling drum simultaneously. IF staining was performed in day 2, and results were obtained. For those which are IF negative in day 2, second shell vial was further incubated to seven days before harvest. In the next stage, a large batch of samples was used to evaluate on the use of Caco-2 shell vial culture in day 2 and day 7. Lastly, Caco-2 shell vial and conventional culture and LLC-MK2 conventional culture were tested for isolation of hMPV. Results: Compared to Caco-2 conventional culture, recovery rate of shell vial culture was elevated slightly. When experimenting on the effect of incubation in rolling drum, results showed that recovery rate was raised in shell vial with rolling drum in day 2, moreover, the percentage of positive cells were increased significantly (p value < 0.05). Furthermore, in the evaluation of Caco-2 shell vial in day 2 and day 7, 75% of samples were isolated in day 2 while 85% were recovered in day 7. Lastly, in the investigation on recovery of hMPV, 53%, 42% and 17% hMPV positive cases were isolated by Caco-2 shell vial, Caco-2 conventional culture and LLC-MK2 conventional culture respectively. Conclusion: First, although recovery rate by shell vial and conventional culture were similar, turnaround time was reduced from a week to a few days by shell vial culture. Therefore, Caco-2 shell vial culture is a more efficient than Caco-2 conventional culture in isolating respiratory viruses. The study also showed that incubation of shell vial in rolling drum able to increase the number of positive cells. Furthermore, in this study, Caco-2 cells were also shown to be more efficient in isolating hMPV when compare to LLC-MK2 cells. / published_or_final_version / Microbiology / Master / Master of Medical Sciences

Made available in DSpace on 2015-12-10T14:23:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-05-17. Added 1 bitstream(s) on 2015-12-10T14:29:41Z : No. of bitstreams: 1 000850005.pdf: 1236403 bytes, checksum: b1877b4be5ffb104086b39681ebd86a7 (MD5) / As hortaliças são itens indispensáveis na dieta dos seres humanos, atuando como adjuvantes na prevenção e tratamento de doenças. Porém, destaca-se o aumento de surtos de Doenças Transmitidas por Alimentos (DTA) associados ao consumo de produtos hortícolas. O objetivo desse trabalho foi verificar o perfil microbiológico de diferentes tipos de saladas de cozinhas hospitalares. No período de 2010 a 2014, o Serviço de Orientação a Alimentação Pública (SOAP) recebeu 641 amostras de saladas provenientes de hospitais da região de Bauru e Botucatu, onde foram submetidas às análises microbiológicas para a determinação do número mais provável de coliformes a 35°C e 45°C, pesquisa de Salmonella spp e contagem de estafilococos coagulase positiva. Os resultados revelaram que em 30,56% das amostras a contagem de coliformes a 35°C foi maior que 1.100 NMP/g e 12,17% apresentaram coliformes a 45°C acima de 100 NMP/g, limite máximo estabelecido pela legislação brasileira. A prevalência de amostras contaminadas sem tratamento térmico foi de 97,44% e de 2,56% para amostras com tratamento térmico, cozidas ou refogadas. Todas as amostras foram negativas para presença de Salmonella spp e apresentaram contagem de estafilococos coagulase positiva < 1,0X102 UFC/g. Apesar das amostras de saladas não oferecerem risco microbiológico associado à patógenos, deve-se atentar para os micro-organismos indicadores, pois as refeições são servidas a indivíduos hospitalizados / Vegetables crops are indispensable items in human diet, acting as adjuvants in the prevention and treatment of diseases. However, the number of Foodborne Diseases outbreaks associated with their consumption increases. The aim of this study was determine the microbiological profile of different salads from hospital kitchens. In the period between 2010 and 2014 the Public feeding orientation service (SOAP) received 641 samples of salads from hospital kitchens in the region of Botucatu and Bauru. They, where subjected to microbiological analysis to determine the most probable number of coliforms at 35 ° C and 45 ° C, besides Salmonella spp presence and coagulase-positive staphylococci counts. The results reveal that 30.56% of the samples had counts for coliform at 35°C greater than 1.100 MPN/g and 12.17% had MPN/g of coliforms at 45°C above 100 MPN/g, the maximum limit established by Brazilian legislation. The prevalence of samples without heat treatment was 2.56% and 97.44% with heat treatment, cooked or braised. All samples were negative for Salmonella spp count and coagulase positive staphylococci < 1,0x102 UFC/g. Although most salads had an adequate microbiological quality, attention should be paid to the safety indicators as the meals are served to hospitalized individuals

Orientador: Germano Francisco Biondi / Banca: Anee Valéria Mendonça Stachissini / Banca: Kate Aparecida Buzi / Resumo: As hortaliças são itens indispensáveis na dieta dos seres humanos, atuando como adjuvantes na prevenção e tratamento de doenças. Porém, destaca-se o aumento de surtos de Doenças Transmitidas por Alimentos (DTA) associados ao consumo de produtos hortícolas. O objetivo desse trabalho foi verificar o perfil microbiológico de diferentes tipos de saladas de cozinhas hospitalares. No período de 2010 a 2014, o Serviço de Orientação a Alimentação Pública (SOAP) recebeu 641 amostras de saladas provenientes de hospitais da região de Bauru e Botucatu, onde foram submetidas às análises microbiológicas para a determinação do número mais provável de coliformes a 35°C e 45°C, pesquisa de Salmonella spp e contagem de estafilococos coagulase positiva. Os resultados revelaram que em 30,56% das amostras a contagem de coliformes a 35°C foi maior que 1.100 NMP/g e 12,17% apresentaram coliformes a 45°C acima de 100 NMP/g, limite máximo estabelecido pela legislação brasileira. A prevalência de amostras contaminadas sem tratamento térmico foi de 97,44% e de 2,56% para amostras com tratamento térmico, cozidas ou refogadas. Todas as amostras foram negativas para presença de Salmonella spp e apresentaram contagem de estafilococos coagulase positiva < 1,0X102 UFC/g. Apesar das amostras de saladas não oferecerem risco microbiológico associado à patógenos, deve-se atentar para os micro-organismos indicadores, pois as refeições são servidas a indivíduos hospitalizados / Abstract: Vegetables crops are indispensable items in human diet, acting as adjuvants in the prevention and treatment of diseases. However, the number of Foodborne Diseases outbreaks associated with their consumption increases. The aim of this study was determine the microbiological profile of different salads from hospital kitchens. In the period between 2010 and 2014 the Public feeding orientation service (SOAP) received 641 samples of salads from hospital kitchens in the region of Botucatu and Bauru. They, where subjected to microbiological analysis to determine the most probable number of coliforms at 35 ° C and 45 ° C, besides Salmonella spp presence and coagulase-positive staphylococci counts. The results reveal that 30.56% of the samples had counts for coliform at 35°C greater than 1.100 MPN/g and 12.17% had MPN/g of coliforms at 45°C above 100 MPN/g, the maximum limit established by Brazilian legislation. The prevalence of samples without heat treatment was 2.56% and 97.44% with heat treatment, cooked or braised. All samples were negative for Salmonella spp count and coagulase positive staphylococci < 1,0x102 UFC/g. Although most salads had an adequate microbiological quality, attention should be paid to the safety indicators as the meals are served to hospitalized individuals / Mestre

In infected patients, the rapid identification of pathogens is critical. After a long period of slow technological improvement, the microbiology laboratory is now undergoing significant evolution. This work evaluated the contribution of recent MALDI-TOF MS technology in terms of the diagnosis and clinical management of patients and its implementation in the laboratory of tomorrow. The studies were conducted over a 3.5-year period, mostly in the iris public hospital network of Brussels. <p>First, we confirmed the accurate performance of MALDI-TOF MS in the identification of routine isolates, regardless of whether the Biotyper (92.7% correct species identification) or VITEK MS (93.2%) (n=986) commercial system was used, and demonstrated the supremacy of this technology over conventional identification techniques for fastidious bacteria, including Campylobacter and related organisms (98.3%, 72.2% and 79.9% correct species identification by Biotyper, Vitek NH Card and API Campy, respectively; n=234). <p>Second, we showed that the direct MALDI-TOF MS identification of bacteria from positive blood cultures was not only feasible but also led to an 24-h reduction in the time-to-identification. In an adult population, more than 13% of the direct identifications from positive blood cultures resulted in the faster adaptation of the antimicrobial treatment. <p>Third, we demonstrated that MALDI-TOF MS could easily be implemented in a network, which was associated with significant cost savings and reduction in the time-to-identification. Finally, our promising Blastocystis subtyping results suggest that the number of MALDI-TOF MS applications may be increased.<p>In the future, automation of the technique will make its use in clinical laboratories even easier, eliminating the use of conventional identification techniques. Improvement of the preanalytical procedures is also important to make MALDI-TOF MS a suitable instrument for resistance and toxicity mechanism detection and subtyping. <p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished