2005
DOI: 10.1128/aem.71.12.8714-8720.2005
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Microcolony Cultivation on a Soil Substrate Membrane System Selects for Previously Uncultured Soil Bacteria

Abstract: Traditional microbiological methods of cultivation recover less than 1% of the total bacterial species, and the culturable portion of bacteria is not representative of the total phylogenetic diversity. Classical cultivation strategies are now known to supply excessive nutrients to a system and therefore select for fast-growing bacteria that are capable of colony or biofilm formation. New approaches to the cultivation of bacteria which rely on growth in dilute nutrient media or simulated environments are beginn… Show more

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Cited by 197 publications
(152 citation statements)
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“…[29][30][31][32][33][34][35][36][37][38][39][40] Although some novel bacterial species were successfully cultured by varying media and growth conditions, 41 significant departures from conventional techniques were clearly in order, and indeed the new technologies substantially diverged from traditional cultivation methods by adopting single cell and high-throughput strategies, 32,37,38,40 better mimicking the natural milieu, 29,31,34,39 increasing the length of incubation and lowering the concentration of nutrients. 33 High-throughput extinction culturing is based on the dilution of natural communities of bacteria to 1-10 cells per well in low nutrient, filtered marine water.…”
Section: Methods For Growing Uncultured Microorganismsmentioning
confidence: 99%
“…[29][30][31][32][33][34][35][36][37][38][39][40] Although some novel bacterial species were successfully cultured by varying media and growth conditions, 41 significant departures from conventional techniques were clearly in order, and indeed the new technologies substantially diverged from traditional cultivation methods by adopting single cell and high-throughput strategies, 32,37,38,40 better mimicking the natural milieu, 29,31,34,39 increasing the length of incubation and lowering the concentration of nutrients. 33 High-throughput extinction culturing is based on the dilution of natural communities of bacteria to 1-10 cells per well in low nutrient, filtered marine water.…”
Section: Methods For Growing Uncultured Microorganismsmentioning
confidence: 99%
“…In a comparison by 16S rRNA gene sequencing to standard actinobacterial petri plate isolation techniques per-formed on the same samples, the traps gave nearly complete enrichment for filamentous actinobacteria, with greater diversity and number, including isolates of rare groups. In a similar study, Ferrari and coworkers had cultured microcolonies of rare soil bacteria by growing them on filters suspended on soil slurry in the absence of added nutrients (22). Micromanipulation was used to grow one isolate in pure culture, and a follow-up study showed that these microcolonies could be reliably micromanipulated for downstream cultivation (23).…”
Section: Approaches To Culturing the Missing Bacterial Diversitymentioning
confidence: 99%
“…26,29) Microcultivation combined with FISH allows the identification of previously "unculturable" bacteria by fluorescence microscopy. 30,31) Thus microcolony-FISH on selective medium containing antibiotics would be a valuable tool with which to rapidly gain information on the number and phylogenetic affiliation of so far uncultured, antibiotic-resistant bacteria from aquatic environments.…”
Section: Discussionmentioning
confidence: 99%