Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2018-06-13T12:50:57Z No. of bitstreams: 1 texto completo.pdf: 739948 bytes, checksum: e22b7ad0fc203f73756f4e4f2c810e59 (MD5) / Made available in DSpace on 2018-06-13T12:50:57Z (GMT). No. of bitstreams: 1 texto completo.pdf: 739948 bytes, checksum: e22b7ad0fc203f73756f4e4f2c810e59 (MD5) Previous issue date: 2018-03-14 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Biofilmes bacterianos formados em áreas de processamento de alimentos podem se tornar fonte de contaminação cruzada. Dessa forma, torna-se importante a correta manipulação das matérias-primas que chegam até a indústria, além do uso adequado de sanitizantes no processo de higienização de equipamentos e superfícies. Objetivou-se com este estudo avaliar a formação de biofilme e potencial de biotransferência de Pseudomonas aeruginosa em aço inoxidável, em contato com leite integral UHT e caldo de vegetais. E o efeito de três surfactantes, em concentrações abaixo, próxima e acima da concentração crítica de micelas, na redução de biofilmes. Cada meio de cultivo foi inoculado com P. aeruginosa ATCC 15442, distribuídos em poços de microplaca contendo cupons de aço inoxidável AISI 304 # 4 e incubados a 15 °C. O número de células aderidas foi determinado após 24, 48, 72, 96 e 120 h e o potencial de biotransferência a partir de 48 h. Soluções de cloreto de benzalcônio (CB), dodecilbenzeno sulfonato de sódio (DBSS) e Tween 80 foram avaliados sobre os biofilmes formados por 72 h. Cupons de aço, antes e após os tratamentos, foram observados por microscopia confocal de varredura a laser. Verificou-se que, apesar do leite integral conter maior concentração de nutrientes em relação ao caldo de vegetais, não houve diferença na formação de biofilme de P. aeruginosa. O potencial de biotransferência do micro-organismo aos meios de cultivo foi observado durante todo o período avaliado. As soluções surfactantes mostraram-se eficientes na redução do número de células aderidas com o aumento da concentração, principalmente o CB. Tween 80 e DBSS reduziram menos de 2 log UFC∙cm -2 de células viáveis, enquanto o CB reduziu até 6 log UFC∙cm -2 . A partir dos resultados obtidos verificou-se que P. aeruginosa pode produzir biofilmes em superfície utilizada no processamento de alimentos, independente do tipo de resíduo alimentar presente. Os surfactantes aplicados apresentaram melhor eficiência em concentrações mais elevadas, acima da crítica de micelas, e a classe desses agentes também contribuiu para a redução das células aderidas. / Bacterial biofilms formed in food processing areas can become a source of cross-contamination. Thus, the correct handling of the raw materials coming into the industry becomes important, as well as the proper use of sanitizing in the cleaning process of equipment and surfaces. The aim of this study was to evaluate the biofilm formation and biotransferential potential of Pseudomonas aeruginosa in stainless steel, in contact with UHT whole milk and vegetable broth. Also, it was evaluated the effect of three surfactants at concentrations below, above and near the critical micelle concentration, in the reduction of biofilms. Each culture medium was inoculated with P. aeruginosa ATCC 15442, distributed in microplate wells containing stainless steel coupons AISI 304 # 4 and incubated at 15 °C. The number of adhered cells was determined after 24, 48, 72, 96 and 120 h and the potential of biotransference after 48 h. Benzalkonium chloride (BAC), sodium dodecylbenzene sulfonate (SDBS) and Tween 80 solutions were evaluated on biofilms formed for 72 h. Steel coupons, before and after treatments, were observed by confocal laser scanning microscopy. It was verified that, although the milk had higher concentration of nutrients in relation to the vegetable broth, there was no difference in the formation of P. aeruginosa biofilm. The biotransfer potential of the microorganism to the culture media was observed throughout the evaluated period. The surfactant solutions were efficient in reducing the number of cells adhered with the concentration increase, mainly of the BAC. Tween 80 and SDBS reduced less than 2 log CFU∙cm -2 of viable cells, while BAC reduced up to 6 log CFU∙cm -2 . From the results obtained it was verified that P. aeruginosa can produce biofilms on the surface used in food processing, regardless of the type of food residue present. The applied surfactants presented better efficiency at higher concentrations, above the critical micelle, and the class of these agents also contributed to the reduction of the adhered cells.

Submitted by Biblioteca da Faculdade de Farmácia (bff@ndc.uff.br) on 2017-03-24T17:22:23Z No. of bitstreams: 1 Deutsch, Gabriela [Dissertação, 2014].pdf: 1124546 bytes, checksum: 4c2efe5d127b5965bea25e458da2afc4 (MD5) / Made available in DSpace on 2017-03-24T17:22:23Z (GMT). No. of bitstreams: 1 Deutsch, Gabriela [Dissertação, 2014].pdf: 1124546 bytes, checksum: 4c2efe5d127b5965bea25e458da2afc4 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A balneoterapia é um importante procedimento realizado diariamente em pacientes queimados. Esta prática consiste na limpeza mecânica com fricção manual sobre as áreas lesionadas pela queimadura utilizando antisséptico. Poucas são as evidências de que sua prática seja efetiva na higienização das feridas e na prevenção de infecção cruzada entre pacientes que utilizam o mesmo tanque. Neste projeto, buscou-se estudar a influência da balneoterapia na descolonização da superfície corporal queimada (SCQ) de pacientes internados em um centro de tratamento de queimaduras (CTQ), quanto à presença de cepas de Staphylococcus aureus e Pseudomonas aeruginosa. Swabs foram coletados durante a realização da balneoterapia de 18 pacientes por 14 semanas. A água utilizada também foi avaliada. Testes fenotípicos e genotípicos foram utilizados para identificação de S. aureus e P. aeruginosa. A susceptibilidade a antimicrobianos e biocidas foi verificada segundo os critérios do Clinical and Laboratory Standards Institute (CLSI). Pulsed-field gel electrophoresis (PFGE) foi utilizado para avaliar a diversidade genômica. Análise exploratória das variáveis envolvidas no processo da balneoterapia foi determinado pela estatística descritiva e testes estatísticos não paramétricos foram utilizados na análise dos fatores de risco. Trezentos e cinquenta e dois swabs foram coletados dos quais 214 (61%) da SCQ, 60 (17%) da cavidade nasal e 78 (22%) da mesa onde ocorreu a balneoterapia. Detectou-se 13 cepas de S. aureus e 39 de P. aeruginosa. A concentração mínima inibitória (CMI) para sulfadiazina de prata foi ≥32μg/mL para as cepas de S. aureus enquanto que para a P. aeruginosa a maioria das cepas apresentou CMI de 128μg/mL. Com relação a clorexidina, as cepas de S.aureus apresentaram um CMI variando de 2 a 8g/mL enquanto para P. aeruginosa a variação foi de 16 a 64μg/mL. Cinco amostras foram identificadas como S. aureus resistentes a meticilina (MRSA) e nove como P. aeruginosa resistentes a carbapenêmicos. A análise do perfil de fragmentação do DNA total (PFGE) nas cepas de P. aeruginosa demonstrou a existência de 10 clones entre 35 cepas analisadas. O tipo A foi o mais prevalente, com 23 cepas distribuídas em 8 subtipos. Estes estavam presentes na SCQ coletada antes e após o banho e nas superfícies da mesa de banho, sugerindo que há contaminação cruzada de um indivíduo para o outro, de uma área queimada para outra no mesmo indivíduo, da mesa da balneoterapia para indivíduos e finalmente do indivíduo para mesa. Os resultados não se mostraram estatisticamente significativos, no entanto, quatro pacientes que não apresentaram contaminação antes do banho se tornaram positivos após este procedimento, e 10 pacientes que apresentaram contaminação antes do banho, assim permaneceram. Conclui-se que o procedimento de descontaminação não está sendo eficaz uma vez que houve similaridade clonal entre as cepas de P. aeruginosa coletadas em vários pontos e momentos / Balneotherapy is an important procedure usually performed in burn patients. This practice consists on a mechanical cleaning with manual friction on the damaged areas using an antiseptic. There is little evidence that this practice is effective to clean the wounds and avoid cross infection between patients using the same table. In this project, we study the influence of hydrotherapy in the decolonization of burned body surface area (BSA) of patients admitted to a burn center (BC) for the presence of Staphylococcus aureus and Pseudomonas aeruginosa isolates. Thus, swabs were used to collect bacteria from 18 patients submitted to balneotherapy during 14 weeks. The material from bath table and the water used were also evaluated. Genotypic and phenotypic tests were used to identify S. aureus and P. aeruginosa. Susceptibility to antimicrobials and biocides has been verified according to the criteria of the Clinical and Laboratory Standards Institute (CLSI). Genomic diversity was assessed by pulsed-field gel electrophoresis (PFGE). Descriptive statistics were used in the exploratory analysis of the variables involved in the balneotherapy and non-parametric statistical tests were used in process analysis of risk factors. Three hundred fifty-two swabs were collected of which 214 (61%) were from BSA, 60 (17%) from nasal cavity and 78 (22%) from table where balneotherapy occurred. Thirteen isolates were identified as S. aureus and 39 as P. aeruginosa. Minimum inhibitory concentration (MIC) for silver sulfadiazine was ≥ 32μg/mL for S. aureus isolates and 128μg/mL for P. aeruginosa. It is possible that the increased MIC to silver sulfadiazine has occurred by the constant use of this antimicrobial in balneotherapy. However, MIC to chlorhexidine for S. aureus isolates range from 2 to 8mg/mL and for P. aeruginosa range from 16 to 64μg/mL. Furthermore, five S. aureus isolates were identified as methicillin-resistant Staphylococcus aureus (MRSA) and nine as P. aeruginosa resistant to carbapenems. A profile analysis of total P. aeruginosa DNA fragmentation showed 10 clones among 35 strains analyzed. Type A was the most prevalent, with 23 isolates distributed into eight subtypes. These subtypes were present in BSA collected before and after the bath and on the surfaces of the bath table, suggesting that there is cross-contamination from one individual to another, from a burned area to another in the same individual, from the balneotherapy table to an individual and finally from the individual to the table. The results were not statistically significant, however, four patients who were not contaminated before bathing became positive after this procedure, and 10 patients who were contaminated before bathing, remained so. Thus, it is possible to conclude that the procedure is not efficient for the decontamination because there was similarity between the clonal isolates of P. aeruginosa collected at various points and times

Current smoking cessation guidelines recommend nicotine replacement therapy (NRT) to assist pregnant smokers to quit, but this is without strong evidence for effectiveness and safety. Nicotine, the main addictive component of tobacco, is known to exert physiological effects by binding to its receptor, the nicotinic acetylcholine receptor (nAChR). Recent studies have identified the presence of nAChRs in non-neuronal cells, and in macrophages, functional alteration upon stimulation with nicotine has been documented. To understand the impact of in utero nicotine exposure on various immune cell disposition and function, we designed preliminary studies using an in vivo model of P. aeruginosa infection. In this model, pregnant mice were exposed to nicotine and after weaning, offspring were infected intra-tracheally and humanely killed 5 days later. Nicotine-exposed mice had a greater weight reduction post-infection. This was accompanied by a decreased number of neutrophil, resident macrophages, and B lymphocytes in the lungs, while the number of B lymphocytes in the lymph nodes were greater than that of the control group. In the lung lavage fluids, IL-6, MCP-1, and TNFα concentrations were elevated in nicotine-exposed mice. In an in vitro system using bone marrow-derived macrophages, a significantly reduced production of IFNγ was observed in nicotine-exposed mice when cells were stimulated with LPS. To characterize and compare gene expression in macrophages isolated from neonates developmentally exposed to nicotine, we designed a clinical study to recruit pregnant mothers who 1) did not smoke during pregnancy, 2) smoked throughout pregnancy, or 3) used NRT during pregnancy. We found that successful RNA isolation can be achieved from neonatal tracheal aspirate samples and cell number and reagent volumes were important determinants of acceptable RNA quality and quantity. Together, these preliminary findings demonstrate a possible alteration in immune response as a result of in utero nicotine exposure and sets a groundwork for future studies in identifying mechanisms underlying the impact of developmental nicotine exposure.

Resistant bacteria are becoming more and more of a problem but treatment with antibiotics in combination may overcome and prevent resistance development. The combination of meropenem and colistin against Pseudomonas aeruginosa has been proposed as a promising treatment to increase the bactericidal effect and a synergistic effect has been proposed. A Pharmacokinetic-Parmacodynamic (PKPD) model that describes the dynamics of bacteria kill could be used to evaluate if the effects are additive or not. The model could later also be used to find optimal dosing for both of the antibiotics used alone or in combination with each other. The aim of the present study was to develop a PKPD model that describes the bactericidal activity of the two antibiotics, both in mono-therapy and in combination. The data were from in vitro static time kill-curve experiments that had been conducted on two strains of Pseudomonas aeruginosa; the wild-type (ATCC 27853) and the resistant-type (PL0603761). Resistance was observed in the experimental data and thus it had to be taken into account in the modelling. PKPD models were fitted to the bacterial counts in NONMEM with pharmacodynamic compartments for susceptible and resting bacteria. In the resting compartment the bacteria could not be killed. The bacteria moved into the resting compartment from the susceptible compartment when a certain concentration of bacteria was obtained. A pharmacokinetic compartment characterized changes in drug concentrations and the drug degradation during the experimental time was considered. Two different drug effects were tried on the susceptible bacteria, linear effect and Emax models.. The resistance development occurring during the experiments was described by two compartments where the parameter kon determined the rate of onset of resistance development. In the final model, kon was found to either be concentration-independent or dependent, depending on antibiotics and bacteria. The degree of resistance development produced an overall inhibitory effect on the drug effect. The growth rate was estimated to be lower and the EC50 to be higher for the resistant compared to the wild-type bacteria. The model was used to predict the expected time-kill curve if the effect of the two drugs are additive when combining the two drugs. The observed  bacteria kill was lower than the model predicted for the wild-type bacteria. For the resistant bacteria the assumption of additive bacteria kill for the two drugs-seemed adequate.

Multidrug resistant bacteria (MDR) have the potential to push back society to the pre-antibiotic era. Although discovered before penicillin, the inexorable rise in antibiotic resistance has revitalised interest in bacteriocins as treatments for bacterial infections. Bacteriocins are protein antibiotics principal to competition amongst pathogens and commensals, but the mechanisms by which they translocate across the Gram-negative cell envelope are poorly understood. The work presented in this thesis demonstrates how the endonuclease bacteriocin pyocin S2 (pyoS2) exploits the iron transporter FpvAI to translocate across the outer membrane (OM) of Pseudomonas aeruginosa. FpvAI is a 22-strand β-barrel and virulence factor in P. aeruginosa that transports iron into the cell in the form of a small siderophore, ferripyoverdine (Fe-Pvd). Uptake of Fe-Pvd requires the proton motive force (PMF), which is transduced to the ligand-bound receptor by TonB1 and its partner proteins ExbB-ExbD in the inner membrane (IM). The crystal structure of the high affinity complex (Kd = 240 pM) formed between the N-terminal domain of pyoS2 (pyoS2^NTD) and FpvAI is presented, which shows pyoS2^NTD mimics Fe-Pvd, and induces the same conformational changes in the receptor. Fluorescently-labelled pyoS2^NTD was actively imported into P. aeruginosa PAO1 cells and this import was dependent on the PMF, TonB1 and a TonB1-box motif at the N-terminus of pyoS2^NTD. Finally, photo-activated crosslinking of stalled translocation intermediates demonstrated pyoS2^NTD translocates through the FpvAI β-barrel lumen by a process analogous to that of Fe-Pvd. Following binding to FpvAI, translocation begins by the unfolding of a force-labile portion of the plug domain, opening a narrow channel through FpvAI. This enables pyoS2 to deliver its own TonB1-box to the periplasm where contact with TonB1 activates its import through the same channel, most likely as an unfolded polypeptide. Hence, this study demonstrates that bacteria possess a rudimentary protein import system that exploits energised nutrient transporters in the OM.

Biofilms are surface-bound bacterial colonies that are held together by a self-produced extracellular polymeric matrix. They are highly resistant to antibiotics and host defence mechanisms, and are known to be the cause of persistent infections. Biofilm-degrading enzymes have been shown to prevent biofilm formation, remove mature biofilm, and enhance the efficacy of antibiotics. This study investigated the antibacterial and antibiofilm actions of the commercial enzyme Pectinex Ultra SP-L (Pectinex), alone and in combination with antibiotics, on standard and clinical cultures of Staphylococcus aureus and Pseudomonas aeruginosa. The cytotoxicity of Pectinex was determined on human cell cultures in vitro. Pectinex (7.42 – 950 PGU/ml) was not bactericidal, and had no effect on the antibacterial efficacy of amoxicillin-clavulanate and ciprofloxacin in cultures of S. aureus (ATCC 12600) and P. aeruginosa (ATCC 9027), respectively. However, in clinical cultures of P. aeruginosa, Pectinex caused an 89.0% (from 1.0 to 1.89 μg/ml) and 92.8% (from 1.67 to 3.22 μg/ml) increase in the MIC and MBC of ciprofloxacin, respectively. In clinical cultures of S. aureus, both bactericidal indices of amoxicillin-clavulanate were increased by 28.0% (from 2.0 to 2.56 μg/ml). In all bacterial cultures, low concentrations of Pectinex (≤ 118.75 PGU/ml) and prolonged incubation periods (≥ 6 h) were both associated with increased viability and biofilm biomass. Over a short incubation period (≤ 6 h), higher concentrations of Pectinex (237.5 – 950 PGU/ml) effectively inhibited biofilm formation in P. aeruginosa ATCC (237.5 – 950 PGU/ml) and clinical (950 PGU/ml) strains but not in S. aureus cultures. Pectinex (237.5 – 950 PGU/ml) was cytotoxic to HeLa cells, lymphocytes and neutrophils, and induced morphological features that included shrunken rounded cells, blebs, apoptotic bodies, cytoplasmic vacuoles and cell debris. The effects at 475 and 950 PGU/ml were comparable to mitomycin C 10 μg/ml and staurosporine 1 μg/ml. Pectinex was shown to either enhance or reduce biofilm biomass and cell viability in cultures of S. aureus and P. aeruginosa. The manifested effects depended on the concentration of the enzyme, the specific bacterial species and strain, and the maturity of the biofilms. Further studies are still needed in order to determine the actions of Pectinex on other clinical pathogens. / Thesis (PhD)--University of Pretoria, 2014. / lk2014 / Pharmacology / PhD / Unrestricted

M.Cur / According to data from the Centers for Disease Control and Prevention’s(CDC) National Nosocomial Infections Surveillance System of 1996, Pseudomonas aeruginosa(P. aeruginosa) can be rated as the number two cause of nosocomial pneumonia(Chen & Rudoy,2006). Nosocomial pneumonia increases hospital cost and morbidity and mortality in patients. Most of the patients in the critical care unit are immune compromised because of underlying illnesses. Antibiotics eliminates the patient’s normal flora which causes opportunity for pathogens to colonise. Indwelling procedures like endotracheal intubation cause a point of entrance for pathogens like P.aeruginosa. The endotracheal tube bypasses the normal physiological processes and inhibits the cough reflex. It is the nurse’s responsibility to remove secretion through endotracheal suctioning. During the past ten years the closed suction method was increasingly implemented to remove secretions because studies showed closed suction caused less infection than open suction. In a spesific critical care unit in a private hospital in Pretoria the nurses are of the opinion that closed suctioning does not effectively remove secretion. Patients are therefore suctioned open which can cause air contamination because the colonised ventilator circuit is opened. The following question can be asked in view of the above arguments and problem statement: Is there a difference in aircontamintion between open and closed suctioning? The aim of the study is to determine whether any difference in air contamination exists between open and closed suctioning in a spesific critical care unit in Pretoria. v A comparitive contextual design with crossover methods was used. Patients are allocated to group 1 or group 2 through random sampling. An air exstractor is used to take airsamples before, during and after suctioning. There was no significant difference in terms of air contamination for open and closed suction. This is probably because of too small a sample. The null hypothesis is accepted and that is there is no significant difference in air contamination between open and closed suction.

Le monitoring des écosystèmes terrestres nécessite une connaissance approfondie des interactions entre microorganismes, minéraux et métaux dans les sols. Afin d'évaluer in vivo la disponibilité de métaux tel que le fer dans des systèmes bactéries-minéraux, une approche basée sur l’utilisation de biosenseurs bactériens fluorescents et d’une analyse spectroscopique non-invasive a été explorée. Ce travail a notamment conduit à la construction chez Pseudomonas aeruginosa de fusions génétiques couplant des promoteurs régulés par le fer à des rapporteurs fluorescents multicolores. Les souches obtenues ont été utilisées comme senseur de la disponibilité du fer constitutif de différents minéraux (Nontronites). La réponse de ces biosenseurs bactériens a été étudiée en couplant la spectroscopie de fluorescence à balayage synchrone (SFS) à la décomposition canonique polyadique Candecomp / Parafac (CP). Avec des plans d’expérience privilégiant la diversité des réponses, le couplage SFS-CP garantit une estimation conjointe et rapide de l’expression de plusieurs promoteurs d’intérêts, y compris dans des milieux auto-fluorescents. Cette méthode originale permet, entre autres, de s’affranchir des problèmes liés aux recouvrements spectraux des protéines fluorescentes et fournit une estimation robuste et précise de la réponse des biosenseurs. Appliquée à d’autres plans d’expériences, elle démontre également que la bio-dissolution des nontronites par P. aeruginosa est assurée par la production de sidérophores et contrôlée par la cristallochimie des feuillets des smectites, notamment par la distribution des atomes de fer(III) entre les tétraèdres et les octaèdres / Monitoring terrestrial ecosystems requires a better understanding of the interactions between microorganisms, minerals and metals in the environment. To assess in vivo availability of metals such as iron in bacteria-mineral system, an approach based on whole-cell fluorescent biosensors and non-invasive spectroscopy was explored. This work led to the construction in Pseudomonas aeruginosa of a set of gene fusions coupling iron-regulated promoters to multicolour fluorescent reporters. The recombinant strains were used as sensors of structural iron availability in nontronites NAu-1 and NAu-2. The response of these biosensors was studied by coupling synchronous fluorescence spectroscopy (SFS) with canonical polyadic Candecomp/Parafac (CP) decomposition. On the basis of experimental designs favouring response diversity, the coupled SFS-CP method guarantees a joint estimate of gene expression from multiple promoters, even in highly fluorescent media. This novel method can solve the issue of spectral bleed-through of fluorescent proteins and provides a means to integrate multiple signals from combinations of whole-cell fluorescent bioreporters. In addition, we could show using SFS-CP that P. aeruginosa indirectly mobilize Fe(III) from nontronites primarily through the production of pyoverdine siderophore. The structural Fe(III) present on the edges of NAu-2 rather than NAu-1 particles appears to be more bioaccessible, suggesting that the distribution of Fe, in the tetrahedron and/or in the octahedron sites, governs the solubilization process

Kött är skelettmuskler från olika djur som till exempel gris, nötdjur eller får. På kött kan många bakterier tillväxa. Pseudomonas är ett släkte bakterier som vanligen orsakar att mat blir dålig. De finns i vår omgivning och kan ge problem bland annat på grund av biofilmbildning inom sjukvård och på livsmedelsindustrier. Slakteriet KLS Ugglarps har tidigare sett att vissa styckningsdetaljer av gris blivit missfärgade med en blå färg och ville ta reda på orsaken till detta. Berörda delar var främst karré och kotlett. Pseudomonas aeruginosa hade tidigare hittats i lokalerna och misstänktes även i detta fall.Kött, både med och utan missfärgningar, undersöktes med hjälp av olika mikrobiologiska metoder för att se vilken bakterie som var orsaken till den blå färgen. Renodling och isolering utfördes och analyser gjordes med hjälp av API 20NE, gramfärgning och oxidastest för att kunna identifiera bakterien och ett referensisolat användes som kontroll. Det visade sig att det inte var P. aeruginosa utan istället en Pseudomonas fluorescens och denna kunde säkerställas som orsaken till blåfärgen genom att den isolerade bakterien från köttet ympades till sterilt kött och då gav en blå färg igen efter inkubering. En smittspårning utfördes i produktionslokalen för att se om bakterien kunde hittas innan uppstart samt under produktionens gång. Proverna visade vid odling att det fanns ytor i lokalen som var odlingspositiva för Pseudomonas. / Meat is skeletal muscle from different animals, such as pigs, cattle or sheep. Pseudomonas are bacteria that may cause food spoilage. The bacteria live in our environment and can cause problems due to biofilm formation in hospitals and industries. The slaughterhouse KLS Ugglarps has found that some pig cuttings have become discolored with at blue color and they wanted to find out what caused it. Pseudomonas aeruginosa had previously been found in the production area and was suspected as the cause.Meat, with and without discoloration, was investigated using various microbiological methods to see which bacterium cause the blue color. Different colonies were isolated and identified. The methods used were API 20NE, gram staining and oxidation test. A reference isolate was used as control. It was found that it was not P. aeruginosa but instead Pseudomonas fluorescens that caused the blue color. This was confirmed by applying the isolated bacteria to sterile meat and the blue color did appear after incubation. A screening for the source of contamination was performed in the production area to see if the bacterium could be found before start-up and during production. The samples taken showed that there were bacteria at both time points and that the production surfaces at the beginning of the production line were more prone to contamination than the other surfaces.

Please read the abstract in the section 00front of this document / Thesis (DPhil (Medical Immunology))--University of Pretoria, 2007. / Immunology / unrestricted