An Improved Method for Viable Counts of Bacteria of the Ovine Rumen which ferment Carbohydrates

Kistner, A.

doi:10.1099/00221287-23-3-565

Cited by 38 publications

(24 citation statements)

References 12 publications

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“…These were adapted from those used by Bryant & Burkey ( I 953) and Kistner (1960). Anaerobic conditions were obtained by purging with oxygen-free gases.…”

Section: Methodsmentioning

confidence: 99%

Characteristics of Cellulolytic Cillobacteria from the Rumens of Sheep Fed Teff (Eragrostis tef) Hay Diets

Gylswyk

Hoffman

1970

Journal of General Microbiology

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“…These were adapted from those used by Bryant & Burkey ( I 953) and Kistner (1960). Anaerobic conditions were obtained by purging with oxygen-free gases.…”

Section: Methodsmentioning

confidence: 99%

Characteristics of Cellulolytic Cillobacteria from the Rumens of Sheep Fed Teff (Eragrostis tef) Hay Diets

Gylswyk

Hoffman

1970

Journal of General Microbiology

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“…Roll bottles of 7 ml. capacity, spun mechanically, were used as culture vessels (Kistner, 1960). Colonies of cellulolytic bacteria were counted after incubation for 4 weeks at 39".…”

Section: Methodsmentioning

confidence: 99%

“…In the 'direct' method as used for instance by Hungate (1947Hungate ( , 1950Hungate ( , 1957, Kistner (1960) ~ Gilchrist & Kistner (1962) ~ Kistner, Gouws & Gilchrist (1962) ' Gilchrist (1965) and De Wet (1966), colony counts of cellulolytic bacteria are made on a selective medium containing finely divided cellulose. The cellulolytic colonies were identified by the circular clearings formed through the action of cellulase secreted by the bacteria.…”

mentioning

confidence: 99%

A Comparison of Two Techniques for Counting Cellulolytic Rumen Bacteria

Gylswyk

1970

Journal of General Microbiology

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S U M M A R YCellulolytic bacteria were counted by the ' direct' method; estimated from the number of clearings produced in films of cellulose-containing agar medium inoculated with high dilutions of rumen ingesta and by the 'indirect' method; numbers were calculated from 'total culturable' counts on films of a non-specific agar medium and from the percentages of bacteria found to be cellulolytic after isolation in pure culture. The two methods yielded similar results.

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“…Measurements of growth of bacteria on their insoluble substrates are problematic for several reasons. Viable counts on plates of agar media (35)(36)(37) are complicated by the propensity of cellulose-decomposing cells to adhere to their substrate, frustrating efforts to accurately dilute bicolor (WT sorgum), or reduced-lignin S. bicolor double mutant (bmr-6 bmr-12 sorghum). Cultures were prepared for microscopy using the dualstaining procedure.…”

Section: Visualization Of C Phytofermentans Cells Growing On Insolublementioning

confidence: 99%

Direct Image-Based Enumeration of Clostridium phytofermentans Cells on Insoluble Plant Biomass Growth Substrates

Alvelo-Maurosa

Lee

Hazen

et al. 2016

Appl Environ Microbiol

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A dual-fluorescent-dye protocol to visualize and quantify Clostridium phytofermentans ISDg (ATCC 700394) cells growing on insoluble cellulosic substrates was developed by combining calcofluor white staining of the growth substrate with cell staining using the nucleic acid dye Syto 9. Cell growth, cell substrate attachment, and fermentation product formation were investigated in cultures containing either Whatman no. 1 filter paper, wild-type Sorghum bicolor, or a reduced-lignin S. bicolor double mutant (bmr-6 bmr-12 double mutant) as the growth substrate. After 3 days of growth, cell numbers in cultures grown on filter paper as the substrate were 6.0-and 2.2-fold higher than cell numbers in cultures with wild-type sorghum and double mutant sorghum, respectively. However, cells produced more ethanol per cell when grown with either sorghum substrate than with filter paper as the substrate. Ethanol yields of cultures were significantly higher with double mutant sorghum than with wild-type sorghum or filter paper as the substrate. Moreover, ethanol production correlated with cell attachment in sorghum cultures: 90% of cells were directly attached to the double mutant sorghum substrate, while only 76% of cells were attached to wild-type sorghum substrate. With filter paper as the growth substrate, ethanol production was correlated with cell number; however, with either wild-type or mutant sorghum, ethanol production did not correlate with cell number, suggesting that only a portion of the microbial cell population was active during growth on sorghum. The dual-staining procedure described here may be used to visualize and enumerate cells directly on insoluble cellulosic substrates, enabling in-depth studies of interactions of microbes with plant biomass. Microbial decomposition of plant biomass is central to nutrient cycling in numerous varied environments, and this process plays a key role in the cycling of carbon on the planet. In terrestrial environments, carbon storage occurs primarily in forests where soils are estimated to store 1,200 gigatons of carbon, approximately two thirds more carbon than found in the atmosphere (1). A major thrust of terrestrial microbial ecology is centered on understanding the composition and function of microbial communities in order to assess their influence on carbon cycling (2). In anoxic environments rich in decaying plant material, diverse communities of interacting microbes are responsible for the degradation of cellulose, complex polysaccharides, and other abundantly produced plant cell wall components. Given that most of these substrates are insoluble, microbial decomposition of plant biomass occurs extracellularly, and breakdown products may become available to other community members, forming a basis for multifarious interactions that occur in these environments (3).Anoxic decomposition of plant biomass is tied to health and nutrition in animals by way of gastrointestinal tract microbial communities. Most animals lack the enzymatic capacity required to digest cellulose and m...

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An Improved Method for Viable Counts of Bacteria of the Ovine Rumen which ferment Carbohydrates

Cited by 38 publications

References 12 publications

Characteristics of Cellulolytic Cillobacteria from the Rumens of Sheep Fed Teff (Eragrostis tef) Hay Diets

Characteristics of Cellulolytic Cillobacteria from the Rumens of Sheep Fed Teff (Eragrostis tef) Hay Diets

A Comparison of Two Techniques for Counting Cellulolytic Rumen Bacteria

Direct Image-Based Enumeration of Clostridium phytofermentans Cells on Insoluble Plant Biomass Growth Substrates

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