Dissertations / Theses on the topic 'Colony forming unit assay'

Thesis (Ph.D. in Pharmaceutical Sciences) -- University of Colorado, 2006.
Typescript. Includes bibliographical references (leaves 191-216). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;

Le système murin est utilisé pour étudier les mécanismes de régulation du choix de différenciation que fait la cellule souche pluripotente hématopoïétique (CFU-S) vers l’une ou l’autre des lignées sanguines. La méthode employée pour cette étude est basée sur l’analyse histologique des nodules spléniques formés par le développement des clones de CFU-S dans la rate de souris irradiée ayant reçu de la moelle 10 jours auparavant. Les proportions de chaque catégorie histologique étant définies et constantes pour une souche de souris donnée, il apparait que tout changement des proportions des différentes catégories histologiques à la suite d’une agression infligée au système hématopoïétique reflète nécessairement une modification de la détermination des cellules souches pluripotentes. L’étude des effets de cytosine arabinoside, administrée sous forme fractionnée à une souris, montre que ce protocole de traitement, in vivo, oriente préférentiellement la détermination des CFU-S médullaires et spléniques vers la granulopoïèse et la mégacaryocytopoïèse au détriment de l’érythropoïèse. De plus, le sérum de souris ainsi traité contient des facteurs humoraux capables d’orienter, in vitro, la détermination de CFU-S réceptrices de moelle de souris normale vers la lignée granulocytaire : ces facteurs, appelés pluripoïétines G, sont sécrétés par la rate et probablement par d’autres tissus. Ces résultats confirment l’hypothèse d’un contrôle humoral de la détermination des CFU-S, notion déjà établie pour la lignée érythrocytaire dans notre laboratoire. La sécrétion des pluripoïétines spécifiques des lignées érythrocytaire et granulocytaire est soumise à un mécanisme de rétrocontrôle qui dépend, tout au moins en partie, des proportions relatives des précurseurs unipotents de chacune de ces lignées
Studies have been carried out in mice to investigate mechanisms of cell differenciation, in particular the determination of the pluripotent haemopoietic stem cell (CFU-S) towards one cell lineage. The method employed is the histological analysis of spleen colonies formed by stem cells 10 days after injection into irradiated mice. Changes in the proportions of colonies of different types as the result of perturbation of haemopoiesis must necessarily reflect a change in the determination of the pluripotent stem cells. A study of the effect of repeated administration of cytosine arabinoside demonstrated a preferential shift in CFU-S determination towards granulopoiesis and megakaryocytopoiesis. In addition, the serum of mice treated in this way contains humoral factors capable, in vitro, of alter determination toward granulopoiesis. These factors have been called pluripoietis G and have been shown to be secreted by the spleen and probably other tissues. These results support the suggestion for a humoral control of CFU-S determination which has been proposed for erythropoiesis in our laboratory. The production of specific pluripoietis for erythroid and granulocytic cell lineages would appear to be under a feedback control at least in part on the relative proportions of unipotents cells in each line of differentiation

This thesis deals with the relationship between HVAC systems and internal microclimate of buildings. The work discusses briefly about different components which are formulating internal microclimate. More details are then focused on the issue of microbial microclimate.

Biologische Systeme sind in der Lage sich an eine Niedrig-Dosis-Bestrahlung anzupassen und eine geringere Sensitivität gegenüber einer nachfolgenden Hoch-Dosis-Bestrahlung zu entwickeln. Dieses Phänomen wird als adaptive Antwort bezeichnet und wurde nach der Bestrahlung mit externen Strahlungsquellen wiederholt in vivo und in vitro untersucht. Im Gegensatz dazu gibt es für die Bestrahlung mit offenen Radionukliden keine systematischen und vergleichenden Untersuchungen. Im Mittelpunkt dieser Arbeit standen der Nachweis sowie die Analyse der adaptiven Antwort an PC Cl3-Zellen nach Bestrahlung mit den offenen Radionukliden Re-188 und Tc-99m. Die Zellschädigung wurde mit dem alkalischen Komet-Assay, zur Bestimmung des initialen DNA-Schadens und dem Koloniebildungstest, zur Ermittlung des klonogenen Überlebens, untersucht. Zur Aufklärung von möglichen Regulationsmechanismen der adaptiven Antwort wurde die Induktion und Reparatur von DSB mit dem gamma-H2AX-Immunfluoreszenz-Assay und die intrazelluläre Radionuklidaufnahme betrachtet. In dieser Arbeit erfolgte erstmals eine systematische Untersuchung der adaptiven Antwort nach Bestrahlung mit offenen Radionukliden in vitro. Insgesamt zeigen diese Ergebnisse, dass nach Bestrahlung mit offenen Radionukliden eine adaptive Antwort ausgebildet wird. Diese ist von der Strahlenqualität während Vor- und Folgebestrahlung sowie der Art der DNA-Schädigung und den initiierten Reparaturprozessen abhängig. Weiter Einflussfaktoren sind die Erholungszeit, die Vorbestrahlung (Dosis, Strahlenqualität) und die Art des Schadensnachweises. Neben den bekannten Regulationsmechanismen wurde erstmals die Reduktion der intrazellulären Radionuklidaufnahme als weitere mögliche adaptive Antwort beschrieben.

The growing demand for strawberries has imposed challenges, especially regarding the control of pests. Many farmers report problems with reduced chemical control efficiency, probably due to selection of resistant populations of insects and mites. An alternative is the use of biological control using pathogenic fungi as a tool in integrated pest management. Metarhizium spp. (Hypocreales: Clavicipitaceae) are generalist entomopathogenic fungi with worldwide distribution and can cause diseases in a large number of hosts. Many studies on the development of Metarhizium as a biological control agent were performed, but this bulk of knowledge is in remarkable contrast to the lack of research on the fundamental ecology of Metarhizium in agroecosystems. This thesis aimed to evaluate the establishment, persistence and dispersal of these entomopathogenic fungi in strawberry crop soil in Inconfidentes, Minas Gerais, Brazil; and to study the diversity and abundance of species of Metarhizium isolated from organic and conventional strawberry crop soils, and the field margins in Brazil and Denmark. The effectiveness of new species of Metarhizium recently found in Brazil, was evaluated against two spotted spider mite, Tetranychus urticae. Applied isolates of M. anisopliae (ESALQ1037) and M. robertsii (ESALQ1426) were able to persist for up to 12 months after the application within the soil, and disperse to other plots and colonize the rhizosphere of strawberry plants. In the plots where ESALQ1037 and ESALQ1426 were applied, 25% and 87.5% of the isolates recovered after 12 months consisted of the same isolates inoculated. A new taxonomically unassigned lineage, referred to as Metarhizium sp. Indet. 5 in this study, was found in strawberry crop margins. The dominant species of Metarhizium in Brazil and Denmark was Metarhizium robertsii and M. brunneum respectively. Further, Metarhizium pemphigi was first detected in Denmark in this study. Soil in organically grown strawberries harbored a more diverse population of Metarhizium spp. compared with conventionally grown strawberries. These studies showed for the first time the potential of new species of Metarhizium as spider mite biological control agents, the lowest median lethal time (LT50 = 4 ± 0.17 days) was observed in mites treated with the isolate ESALQ1638 of Metarhizium sp. indet. 1. The best isolates were ESALQPL63 of B. bassiana, ESALQ1608 and ESALQ1638 of Metarhizium sp. indet. 1 and ESALQ3069 and ESALQ3222 of M. pingshaense based on the survival curve, total mortality, percentage of sporulated cadavers and LT50. Knowledge of the diversity of Metarhizium spp. and persistence in strawberry soil generated in this study may be useful in developing conservation strategies and maximize the natural biological pest control.
A crescente demanda por morangos vem impondo desafios, especialmente quanto ao controle das pragas. Muitos agricultores relatam problemas com a redução da eficiência do controle químico, provavelmente devido à seleção de populações resistentes de insetos e ácaros. Uma alternativa é o uso de controle biológico com fungos entomopatogênicos como ferramenta dentro do manejo integrado de pragas. Metarhizium spp. (Hypocreales: Clavicipitaceae), são fungos entomopatogênicos generalistas com distribuição cosmopolita e que podem causar doenças em um grande número de hospedeiros. Muitos estudos sobre o desenvolvimento de Metarhizium como agente de controle biológico foram realizados, mas este leque de conhecimento está em contraste com a notável falta de investigação sobre a ecologia de Metarhizium nos agroecossistemas. Esta tese teve como objetivo avaliar o estabelecimento, persistência e dispersão destes fungos entomopatogênicos em solo de morangueiro em Inconfidentes, Minas Gerais, Brasil; bem como estudar a diversidade e abundância de espécies de Metarhizium isolados do solo de cultivos orgânico e convencional de morangueiro, e das margens das plantações no Brasil e Dinamarca. A eficácia de novas espécies de Metarhizium, encontradas recentemente no Brasil, foi avaliada contra o ácaro rajado, Tetranychus urticae. Os isolados inoculados de M. anisopliae (ESALQ1037) e M. robertsii (ESALQ1426) foram capazes de persistir por até 12 meses após a aplicação no solo, além de dispersar para outras parcelas e colonizar a rizosfera dos morangueiros. Nas parcelas onde ESALQ1037 e ESALQ1426 foram aplicados, 25% e 87,5% dos isolados recuperados após 12 meses consistiam dos mesmos isolados inoculados. Uma nova linhagem não taxonomicamente identificada, referida neste trabalho como Metarhizium sp. Indet. 5, foi encontrada nas margens de morangueiros cultivados. A espécie dominante de Metarhizium no Brasil e Dinamarca foi Metarhizium robertsii e M. brunneum, respectivamente. Além disso, Metarhizium pemphigi foi detectado pela primeira vez na Dinamarca neste estudo. Solos de cultivo orgânico de morangueiro em geral apresentaram uma diversidade maior de Metarhizium do que solos de cultivos convencionais. Estes estudos revelaram pela primeira vez o potencial de novas espécies de Metarhizium como agentes de controle biológico do ácaro rajado, sendo o menor tempo letal mediano (TL50= 4 ± 0.17 dias) observado em ácaros tratados com o isolado ESALQ1638 de Metarhizium sp. indet. 1. Os melhores isolados foram ESALQPL63 de B. bassiana, ESALQ1608 e ESALQ1638 de Metarhizium sp. indet. 1 e ESALQ3069 e ESALQ3222 de M. pingshaense baseado na curva de sobrevivência, mortalidade total, porcentagem de cadáveres esporulados e TL50. O conhecimento da diversidade de Metarhizium spp. e persistência em solos de morango, gerados neste estudo, poderão ser úteis no desenvolvimento de estratégias de conservação e maximizar o controle biológico natural de pragas.

The history of surgery is nearly as old as the human race. Control of wound infection has always been an essential part of any surgical procedure, and is still an important challenge in hospital operating rooms today. For patients undergoing surgery there is always a risk that they will develop some kind of postoperative complication. It is widely accepted that airborne bacteria reaching a surgical site are mainly staphylococci released from the skin flora of the surgical staff in the operating room and that even a small fraction of those particles can initiate a severe infection at the surgical site.  Wound infections not only impose a tremendous burden on healthcare resources but also pose a major threat to the patient. Hospital-acquired infection ranks amongst the leading causes of death within the surgical patient population. A broad knowledge and understanding of sources and transport mechanisms of infectious particles may provide valuable possibilities to control and minimize postoperative infections. This thesis contributes to finding solutions, through analysis of such mechanisms for a range of ventilation designs together with investigation of other factors that can influence spread of infection in hospitals, particularly in operating rooms. The aim of this work is to apply the techniques of computational fluid dynamics in order to provide better understanding of air distribution strategies that may contribute to infection control in operating room and ward environments of hospitals, so that levels of bacteria-carrying particles in the air can be reduced while thermal comfort and air quality are improved.  A range of airflow ventilation principles including fully mixed, laminar and hybrid strategies were studied. Airflow, particle and tracer gas simulations were performed to examine contaminant removal and air change effectiveness. A number of further influential parameters on the performance of airflow ventilation systems in operating rooms were examined and relevant measures for improvement were identified. It was found that airflow patterns within operating room environments ranged from laminar to transitional to turbulent flows. Regardless of ventilation system used, a combination of all airflow regimes under transient conditions could exist within the operating room area. This showed that applying a general model to map airflow field and contaminant distribution may result in substantial error and should be avoided. It was also shown that the amount of bacteria generated in an operating room could be minimized by reducing the number of personnel present. Infection-prone surgeries should be performed with as few personnel as possible. The initial source strength (amount of colony forming units that a person emits per unit time) of staff members can also be substantially reduced, by using clothing systems with high protective capacity. Results indicated that horizontal laminar airflow could be a good alternative to the frequently used vertical system. The horizontal airflow system is less sensitive to thermal plumes, easy to install and maintain, relatively cost-efficient and does not require modification of existing lighting systems. Above all, horizontal laminar airflow ventilation does not hinder surgeons who need to bend over the surgical site to get a good view of the operative field. The addition of a mobile ultra-clean exponential laminar airflow screen was also investigated as a complement to the main ventilation system in the operating room. It was concluded that this system could reduce the count of airborne particles carrying microorganisms if proper work practices were maintained by the surgical staff. A close collaboration and mutual understanding between ventilation experts and surgical staff would be a key factor in reducing infection rates. In addition, effective and frequent evaluation of bacteria levels for both new and existing ventilation systems would also be important.
Tidigt i mänsklighetens utveckling har kirurgin funnits med i bilden. Hantering av infektioner har genom tiderna varit en oundviklig del av alla kirurgiska ingrepp, och finns kvar ännu idag som en viktig utmaning i operationssalar på sjukhus. För patienter som genomgår kirurgi finns alltid en risk att de efter ingreppet utvecklar någon behandlingsrelaterad komplikation. Allmänt accepterat är att de luftburna bakterier som når operationsområdet huvudsakligen består av stafylokocker frigjorda från hudfloran av operationspersonalen i operationssalen, och att endast en liten del av dessa partiklar behövs för att initiera en allvarlig infektion i det behandlade området. Sårinfektioner innebär inte bara en enorm börda för hälso- och sjukvårdsresurser, utan utgör också en betydande risk för patienten. På sjukhus förvärvad infektion finns bland de främsta dödsorsakerna i kirurgiska patientgrupper.. En bred kunskap och förståelse av spridningsmekanismer och källor till infektionsspridande partiklar kan ge värdefulla möjligheter att kontrollera och minimera postoperativa infektioner. Denna avhandling bidrar till lösningar genom analys av en rad olika ventilationssystem tillsammans med undersökning av andra faktörer som kan påverka infektionsspridningen på sjukhus, främst i operationssalar. Syftet med arbetet är att med hjälp av CFD-teknik (Computational Fluid Dynamics) få bättre förståelse för olika luftspridningsmekanismers betydelse vid ventilation av operationssalar och vårdinrättningar på sjukhus, så att halten av bacteriebärande partiklar i luften kan minskas samtidigt som termisk komfort och luftkvalité förbättras.  Flera luftflödesprinciper för ventilation inklusive omblandade strömning, riktad (laminär) strömning och hybridstrategier har studerats. Simuleringar av luft-, partikel- och spårgasflöden gjordes för alla fallstudier för att undersöka partikelevakuering och luftomsättning i rummet. Flera viktiga parametrar som påverkar detta undersöktes och relevanta förbättringar  föreslås i samarbete med industrin. Av resultaten framgår att mängden genererade bakterier i en operationssal  kan begränsas genom att minska antalet personer i operationsteamet. Infektionsbenägna operationer skall utföras med så lite personal som möjligt. Den initiala källstyrkan (mängden kolonibildande enheter som en person avger per tidsenhet) från operationsteamet kan avsevärt minskas om högskyddande kläder används. Av resultaten framgår också att ett horisontellt (laminärt) luftflöde kan vara ett bra alternativ till det ofta använda vertikala luftflödet. Ett horisontellt luftflöde är mindre känsligt för termisk påverkan från omgivningen, enkelt att installera och underhålla, relativt kostnadseffektivt och kräver vanligen ingen förändring av befintlig belysningsarmatur. Framför allt begränsar inte denna ventilationsprincip kirurgernas rörelsemönster. De kan luta kroppen över operationsområdet utan att hindra luftflödet. En flyttbar flexibel skärm för horisontell spridning av ultraren ventilationsluft i tillägg till ordinarie ventilation undersöktes också. Man fann att denna typ av tilläggsventilation kan minska antalet luftburna partiklar som bär mikroorganismer om operationspersonalen följer en strikt arbetsordning. Bra samarbete och förståelse mellan ventilationsexperter och operationsteamet på sjukhuset är nyckeln till att få ner infektionsfrekvensen. Det är också viktigt med effektiva och frekventa utvarderingar av bakteriehalten i luften, för såväl nya  som befintliga ventilationssystem.

QC 20160129

Accurate quantitative analysis of microorganisms is recognized as an essential tool for gauging safety and quality in a wide range of fields. The enumeration processes of viable microorganisms via traditional culturing techniques are methodically convenient and cost-effective, conferring high applicability worldwide. However, manual counting can be time-consuming, laborious and imprecise. Furthermore, particular pathologies require an urgent and accurate response for the therapy to be effective. To reduce time limitations and perhaps discrepancies, this work introduces an intelligent image processing software capable of automatically quantifying the number of Colony Forming Units (CFUs) in Petri-plates. This rapid enumeration enables the technician to provide an expeditious assessment of the microbial load. Moreover, an auxiliary system is able to differentiate among colony images of Echerichia coli, Pseudomonas aeruginosa and Staphylococcus aureus via Machine Learning, based on a Convolutional Neural Network in a process of cross-validation. For testing and validation of the system, the three bacterial groups were cultured, and a significant labeled database was created, exercising suited microbiological laboratory methodologies and subsequent image acquisition. The system demonstrated acceptable accuracy measures; the mean values of precision, recall and F-measure were 95%, 95% and 0.95, for E. coli, 91%, 91% and 0.90 for P. aeruginosa, and 84%, 86% and 0.85 for S. aureus. The adopted deep learning approach accomplished satisfactory results, manifesting 90.31% of accuracy. Ultimately, evidence related to the time-saving potential of the system was achieved; the time spent on the quantification of plates with a high number of colonies might be reduced to a half and occasionally to a third.
A análise quantitativa de microrganismos é uma ferramenta essencial na aferição da segurança e qualidade numa ampla variedade de áreas. O processo de enumeração de microrganismos viáveis através das técnicas de cultura tradicionais é económica e metodologicamente adequado, conferindo lhe alta aplicabilidade a nível mundial. Contudo, a contagem manual pode ser morosa, laboriosa e imprecisa. Em adição, certas patologias requerem uma urgente e precisa resposta de modo a que a terapia seja eficaz. De forma a reduzir limitações e discrepâncias, este trabalho apresenta um software inteligente de processamento de imagem capaz de quantificar automaticamente o número de Unidades Formadoras de Colónias (UFCs) em placas de Petri. Esta rápida enumeração, possibilita ao técnico uma expedita avaliação da carga microbiana. Adicionalmente, um sistema auxiliar tem a capacidade de diferenciar imagens de colónias de Echerichia coli, Pseudomonas aeruginosa e Staphylococcus aureus recorrendo a Machine Learning, através de uma Rede Neuronal Convolucional num processo de validação cruzada. Para testar e validar o sistema, os três grupos bacterianos foram cultivados e uma significativa base de dados foi criada, recorrendo às adequadas metodologias microbiológicas laboratoriais e subsequente aquisição de imagens. O sistema demonstrou medidas de precisão aceitáveis; os valores médios de precisão, recall e F-measure, foram 95%, 95% e 0.95, para E. coli, 91%, 91% e 0.90 para P. aeruginosa, e 84%, 86% e 0.85 para S. aureus. A abordagem deep learning obteve resultados satisfatórios, manifestando 90.31% de precisão. O sistema revelou potencial em economizar tempo; a duração de tarefas afetas à quantificação de placas com elevado número de colónias poderá ser reduzida para metade e ocasionalmente para um terço.

碩士
國立成功大學
口腔醫學研究所
101
Post-natal stem cells originated from pulp of wisdom teeth may be applied for regenerative medicine. The reason and timing for wisdom teeth removal should be re-evaluated. In the aspect of embryology, teeth develop through bud stage, cap stage, bell stage, and then root formation. Our aim is to compare the quantity of CFU in cells isolated from pulp tissue or germ of human wisdom teeth at different developmental stages. Clinically, we can see the developing teeth in radiographs only until calcification of tissue in the late bell stage. We divided patients' teeth into 5 groups: bell stage (group 1), 1/2 root formation (group 2), 3/4 root formation (group 3), well-formed root but open apex (group 4), well-formed root with closed apex (group 5), according to patients' panoramic films. If patients fit the indication of wisdom teeth removal, teeth were removed by a standard oral surgical procedure and the soft tissues inside the crown were taken. After enzymatic dissociation of these soft tissues, we isolated cells from them and performed the primary cell culture. Positive correlation between CFU and tissue weight was noted. But colony was not observed in the group 5.