Isolation of Typical Marine Bacteria by Dilution Culture: Growth, Maintenance, and Characteristics of Isolates under Laboratory Conditions

Schut, Frits; Vries, E.J. de; Gottschal, Jan C.; Robertson, Betsy R.; Harder, W.; Prins, R. A.; Button, D. K.

doi:10.1128/aem.59.7.2150-2160.1993

Cited by 268 publications

(162 citation statements)

References 82 publications

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“…Sphingopyxis alaskensis is a marine bacterium that was isolated as a numerically abundant species from permanently cold (4-10°C) waters in the Alaskan and Japanese North Pacific, and the North Sea (Schut et al, 1993;1997;Eguchi et al, 1996;2001). Its ability to proliferate to high numbers illustrates its capacity to compete effectively at low temperature, and under all the other biotic and abiotic factors prevailing at the times of sampling.…”

Section: Introductionmentioning

confidence: 99%

Cold adaptation in the marine bacterium, Sphingopyxis alaskensis, assessed using quantitative proteomics

Ting

Williams²,

Cowley

et al. 2010

Environmental Microbiology

123

101

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The cold marine environment constitutes a large proportion of the Earth's biosphere. Sphingopyxis alaskensis was isolated as a numerically abundant bacterium from several cold marine locations, and has been extensively studied as a model marine bacterium. Recently, a metabolic labelling platform was developed to comprehensively identify and quantify proteins from S. alaskensis. The approach incorporated data normalization and statistical validation for the purpose of generating highly confident quantitative proteomics data. Using this approach, we determined quantitative differences between cells grown at 10°C (low temperature) and 30°C (high temperature). Cold adaptation was linked to specific aspects of gene expression: a dedicated protein-folding system using GroESL, DnaK, DnaJ, GrpE, SecB, ClpB and PPIase; polyhydroxyalkanoate-associated storage materials; a link between enzymes in fatty acid metabolism and energy generation; de novo synthesis of polyunsaturated fatty acids in the membrane and cell wall; inorganic phosphate ion transport by a phosphate import PstB homologue; TonB-dependent receptor and bacterioferritin in iron homeostasis; histidine, tryptophan and proline amino acid metabolism; and a large number of proteins without annotated functions. This study provides a new level of understanding on how important marine bacteria can adapt to compete effectively in cold marine environments. This study is also a benchmark for comparative proteomic analyses with other important marine bacteria and other cold-adapted organisms.

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Section: Introductionmentioning

confidence: 99%

Cold adaptation in the marine bacterium, Sphingopyxis alaskensis, assessed using quantitative proteomics

Ting

Williams²,

Cowley

et al. 2010

Environmental Microbiology

123

101

View full text Add to dashboard Cite

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“…It has been suggested that most of the bacterial species are nonplatable because they are adapted to low nutrient concentrations and reach only low maximum growth rates (K-selected) [2,10]. The physiology and ecological role of these unknown bacterial species in most cases can only be assessed after their isolation in pure culture.…”

Section: Introductionmentioning

confidence: 99%

“…These parameters were then used to systematically improve culturability. In addition, we established de¢ned growth media to replace the often used complex media (e.g., R2A [23]), or autoclaved sitewater [2,19].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of cell activity and of methods for the cultivation of bacteria from a natural lake community

Bartscht

1999

FEMS Microbiology Ecology

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The viability and culturability of chemoheterotrophic bacteria in Zwischenahner Meer, a shallow eutrophic lake in Northern Germany, were assessed by different techniques. The fraction of metabolically active cells was measured by the CTC reduction method. At three sampling times, numbers of CTC-reducing cells represented only a minor fraction (98%) of the total cell count. Only a slight stimulation of CTC reduction was observed upon the addition of single carbon substrates or substrate mixtures. A medium mimicking the low natural ion concentrations in freshwater was devised and used for the cultivation of bacteria in most probable number (MPN) series and on membrane filters. Similar to the number of CTC-reducing cells, the number of culturable cells as determined in MPN series never exceeded 7% of the total cell count with any of the carbon substrates tested. This indicated that the type of carbon substrate was not the major determinant of the rather low culturability. However, we observed a significant inhibition of bacterial growth in MPN series in which the typical R2A medium was used, or when phosphate buffer instead of HEPES was employed. In contrast to the low numbers of CTC-reducing cells and the low MPN values, up to 58% of the bacterial cells could divide on membrane filters incubated on top of the same type of medium that had been used in the MPN series. It is concluded that most of the bacteria in Zwischenahner Meer are able to grow and divide, but do not reach high cell densities even in improved culture media. Therefore, the majority of bacteria may be classified as viable but nonculturable, but would not be detected by the CTC reduction method. z

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“…Enrichment of microbial communities that couple anaerobic acetate oxidation to Mn 31 reduction Enrichment cultures were initiated by transferring 2.5 g of homogenized sediment from the Mn 21 -rich redox zone (30-50 mm depth interval) to 100 mL of anaerobic growth medium in serum bottles under a N 2 atmosphere. Growth medium consisted of sterile sulphate-free artificial seawater (Schut et al, 1993) diluted two-fold with anoxic deionized water to mimic salt marsh ionic strength. Replicate enrichments were amended with either 100 mM, 1 mM, or 14 mM sodium sulphate.…”

Section: Sediment Sampling and Field Measurementsmentioning

confidence: 99%

Microbial manganese(III) reduction fuelled by anaerobic acetate oxidation

Szeinbaum

Lin

Brandes

et al. 2017

Environmental Microbiology

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Soluble manganese in the intermediate +III oxidation state (Mn ) is a newly identified oxidant in anoxic environments, whereas acetate is a naturally abundant substrate that fuels microbial activity. Microbial populations coupling anaerobic acetate oxidation to Mn reduction, however, have yet to be identified. We isolated a Shewanella strain capable of oxidizing acetate anaerobically with Mn as the electron acceptor, and confirmed this phenotype in other strains. This metabolic connection between acetate and soluble Mn represents a new biogeochemical link between carbon and manganese cycles. Genomic analyses uncovered four distinct genes that allow for pathway variations in the complete dehydrogenase-driven TCA cycle that could support anaerobic acetate oxidation coupled to metal reduction in Shewanella and other Gammaproteobacteria. An oxygen-tolerant TCA cycle supporting anaerobic manganese reduction is thus a new connection in the manganese-driven carbon cycle, and a new variable for models that use manganese as a proxy to infer oxygenation events on early Earth.

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Isolation of Typical Marine Bacteria by Dilution Culture: Growth, Maintenance, and Characteristics of Isolates under Laboratory Conditions

Cited by 268 publications

References 82 publications

Cold adaptation in the marine bacterium, Sphingopyxis alaskensis, assessed using quantitative proteomics

Cold adaptation in the marine bacterium, Sphingopyxis alaskensis, assessed using quantitative proteomics

Evaluation of cell activity and of methods for the cultivation of bacteria from a natural lake community

Microbial manganese(III) reduction fuelled by anaerobic acetate oxidation

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