Bioluminescence Assay for Measuring the Number of Bacteria Adhering to the Hydrocarbon Phase in the BATH Test

Vanhaecke, Erwin; Pijck, J.

doi:10.1128/aem.54.6.1436-1439.1988

Cited by 23 publications

(12 citation statements)

References 27 publications

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“…Although variable results were found depending on the hydrocarbon, buffer and strain employed, we consider, in agreement with Vanhaecke and Pijck [21], more appropriate the use of hexadecane and PUM buffer, since a higher correlation with the other methods was detected.…”

Section: Discussionsupporting

confidence: 84%

Influence of the growth conditions on the hydrophobicity of Renibacterium salmoninarum evaluated by different methods

Bandín¹

1989

FEMS Microbiology Letters

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The influence of medium and salinity on the cell surface hydrophobicity of Renibacterium salmoninarum was investigated using three different methods: bacterial adherence to hydrocarbons (BATH), salt agglutination test (SAT), and binding to nitrocellulose filters (NCF). The possible relationship among hydrophobicity, haemagglutination and adherence to cell lines was also evaluated. R. salmoninarum showed to be highly hydrophobic regardless of the growth conditions or technique employed. Nevertheless, slight differences can be detected depending on the method used. In the SAT and NCF assays very uniform values were obtained within the strains. All the R. salmoninarum isolates agglutinated in (NH4)2SO4 in a range of 0.05-0.2 M and displayed a 77-100% of adherence to nitrocellulose filters. However, more variable results were observed in the BATH method depending on the hydrocarbon, buffer and strain employed. Although all of the isolates produced haemagglutinins for homeotherm erythrocytes, the majority of them failed to agglutinate poikilothermic red blood cells and were unable to adhere to fish cell lines. Therefore, a general correlation among hydrophobicity, agglutinating capacity for fish erythrocytes and adherence to fish cells can not be established for R. salmoninarum.

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Section: Discussionsupporting

confidence: 84%

Influence of the growth conditions on the hydrophobicity of Renibacterium salmoninarum evaluated by different methods

Bandín¹

1989

FEMS Microbiology Letters

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“…The bacterial adherence to hydrocarbons (BATH) test was performed as described by and Weiss et al (44), using hexadecane and octane as organic test fluids. Xylene was excluded from the test because it was found to produce unreliable hydrophobicity results (38).…”

Section: Measurement Of Bacterial Cell Hydrophobicity (I) Bathmentioning

confidence: 99%

“…(i) Instrumentation and reagents. Light production was measured by using Lumit-PM (luciferin-luciferase reagent) in a Lumac Biocounter M2010 (Lumac bv, Landgraaf, The Netherlands) (20,27,38). Nucleotide-releasing reagent for microbial cells (NRB) (Lumac) was used as the bacterial ATP-releasing agent.…”

Section: Measurement Of Bacterial Cell Hydrophobicity (I) Bathmentioning

confidence: 99%

Kinetics of Pseudomonas aeruginosa adhesion to 304 and 316-L stainless steel: role of cell surface hydrophobicity

Vanhaecke¹,

Remon²,

Moors³

et al. 1990

Appl Environ Microbiol

186

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Fifteen different isolates of Pseudomonas aeruginosa were used to study the kinetics of adhesion to 304 and 316-L stainless steel. Stainless steel plates were incubated with approximately 1.5 x 107 CFU/ml in 0.01 M phosphate-buffered saline (pH 7.4). After the plates were rinsed with the buffer, the number of adhering bacteria was determined by a bioluminescence assay. Measurable adhesion, even to the electropolished surfaces, occurred within 30 s. Bacterial cell surface hydrophobicity, as determined by the bacterial adherence to hydrocarbons test and the contact angle measurement test, was the major parameter influencing the adhesion rate constant for the first 30 min of adhesion. A parabolic relationship between the CAM values and the logarithm of the adhesion rate constants (In k) was established. No correlation between either the salt aggregation or the improved salt aggregation values and the bacterial adhesion rate constants could be found. Since there was no significant correlation between the bacterial electrophoretic mobilities and the In k values, the bacterial cell surface charge seemed of minor importance in the process of adhesion of P. aeruginosa to 304 and 316-L stainless steel.

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“…Many investigators have modified the original MATH test and have found that seemingly small variations in experimental conditions, such as the diameter of the test tubes, the pH of the suspension medium, and the volume of hydrocarbon used, can significantly alter the results (4,17,30). Aliphatic hydrocarbons are used in most cases, since aromatic hydrocarbons cause lysis of some bacterial species (55).…”

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confidence: 99%

Cell Surface Analysis Techniques: What Do Cell Preparation Protocols Do to Cell Surface Properties?

Pembrey

Marshall

Schneider

1999

Appl Environ Microbiol

215

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Cell surface analysis often requires manipulation of cells prior to examination. The most commonly employed procedures are centrifugation at different speeds, changes of media during washing or final resuspension, desiccation (either air drying for contact angle measurements or freeze-drying for sensitive spectroscopic analysis, such as X-ray photoelectron spectroscopy), and contact with hydrocarbon (hydrophobicity assays). The effects of these procedures on electrophoretic mobility, adhesion to solid substrata, affinity to a number of Sepharose columns, structural integrity, and cell viability were systematically investigated for a range of model organisms, including carbon- and nitrogen-limited Psychrobacter sp. strain SW8 (glycocalyx-bearing cells), Escherichia coli(gram-negative cells without a glycocalyx), and Staphylococcus epidermidis (gram-positive cells without a glycocalyx). All of the cell manipulation procedures severely modified the physicochemical properties of cells, but with each procedure some organisms were more susceptible than others. Considerable disruption of cell surfaces occurred when organisms were placed in contact with a hydrocarbon (hexadecane). The majority of cells became nonculturable after air drying and freeze-drying. Centrifugation at a high speed (15,000 × g) modified many cell surface parameters significantly, although cell viability was considerably affected only in E. coli. The type of washing or resuspension medium had a strong influence on the values of cell surface parameters, particularly when high-salt solutions were compared with low-salt buffers. The values for parameters obtained with different methods that allegedly measure similar cell surface properties did not correlate for most cells. These results demonstrate that the methods used to prepare cells for cell surface analysis need to be critically investigated for each microorganism so that the final results obtained reflect the nature of the in situ microbial cell surface as closely as possible. There is an urgent need for new, reliable, nondestructive, minimally manipulative cell surface analysis techniques that can be used in situ.

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Bioluminescence Assay for Measuring the Number of Bacteria Adhering to the Hydrocarbon Phase in the BATH Test

Cited by 23 publications

References 27 publications

Influence of the growth conditions on the hydrophobicity of Renibacterium salmoninarum evaluated by different methods

Influence of the growth conditions on the hydrophobicity of Renibacterium salmoninarum evaluated by different methods

Kinetics of Pseudomonas aeruginosa adhesion to 304 and 316-L stainless steel: role of cell surface hydrophobicity

Cell Surface Analysis Techniques: What Do Cell Preparation Protocols Do to Cell Surface Properties?

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