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1.
J Microbiol Methods ; 63(1): 29-35, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16157210

RESUMO

An effective microbial preservation technology for the long-term storage of viable prokaryotic cells is described. The method combines an almost instantaneous drying step with minimal stress to the cells during drying to maximize survival on rehydration. This is achieved by contact of a microlitre aliquot of a bacterial suspension with a novel, pre-dried activated charcoal cloth based matrix contained within a re-sealable system that can then be stored. The simple methodology completely circumvents the requirement for further drying or preparation. Using this method, a standard laboratory Escherichia coli strain was successfully revived following 390 days storage at 4, 20 and 30 degrees C. Data obtained yielded approximately 20%, 6% and 0.1% viable organisms at the aforementioned temperatures, respectively, following initial inoculations of 1.1 x 10(8)/microl cells. While these figures represent a significant viability loss, there is sufficient recovery of microorganisms required for maintaining culture collections.


Assuntos
Técnicas Bacteriológicas/métodos , Preservação Biológica/métodos , Aliivibrio fischeri , Carvão Vegetal , Escherichia coli , Fatores de Tempo
2.
Biosens Bioelectron ; 19(11): 1457-63, 2004 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-15093217

RESUMO

The design and performance characteristics of a small volume (20 ml) continuous culture device for the cultivation of luminous bacteria are described. This simply constructed device can be used to supply luminescent bacteria with constant properties for either laboratory use or the assay of environmental pollutants. Furthermore, bacteria can be deployed to make sensitive (<1 nM) oxygen measurements. The culture device may be configured, alone or in combination, as a chemostat, turbidostat or a "bioluminostat" where bacterial bioluminescence becomes the controlling variable. Over extended periods (>1 week) it was possible to maintain steady-state luminescence within 5% of a pre-set value, although occasionally a non-bioluminescent "mutant" became dominant; in this case light emission was irreversibly lost. A secondary chamber provided additional flexibility and easy manipulation of the cultivated bacteria for testing. The continuous culture system is also suitable for the growth of recombinant microorganisms that either constitutively express luciferase, or do so in response to stress promoter activity. The non-standard control methodologies reported may have important research and industrial applications, for example in providing immediate process control or as an inferential method to optimize biomass: product yield ratios.


Assuntos
Técnicas de Laboratório Clínico/instrumentação , Photobacterium , Aliivibrio fischeri , Técnicas Biossensoriais , Técnicas de Cultura de Células/instrumentação , Técnicas de Cultura de Células/métodos , Técnicas de Laboratório Clínico/métodos , Oxigênio/metabolismo , Photobacterium/metabolismo , Fatores de Tempo
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