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Egg yolk extract

Manufactured by Thermo Fisher Scientific

Egg yolk extract is a raw material derived from the yolk of chicken eggs. It is a complex mixture of proteins, lipids, and other biomolecules. The core function of egg yolk extract is to serve as a source of these naturally occurring compounds for use in various applications, such as research, manufacturing, and product formulations. This description is factual and unbiased, without interpretation or extrapolation on the intended use of the product.

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3 protocols using egg yolk extract

1

Phospholipase and Gelatinase Activity Assays

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The phospholipase/lecithinase activity was assayed using agar plates supplemented with egg yolk emulsion as a lecithin source. Ten microliters of TSB-1 overnight cultures for each P. damselae subsp. damselae strain were spotted onto TSA-1 plates supplemented with 3% egg yolk extract (Oxoid), and results were evaluated after 24 h of culture at 25°C. Hydrolysis of lecithin by the phospholipase yields water-insoluble diglycerides that cause the appearance of an opaque precipitate. The gelatinase activity assay was carried out by spotting 10 μl of a TSB-1 overnight culture onto TSA-1 plates supplemented with 1% gelatin (Oxoid), and results were developed after 48 h of incubation at 25°C by covering the agar plate surface with a 12.5% (wt/vol) HgCl2 solution. Hydrolysis of gelatin by the gelatinase enzyme causes the appearance of a translucent halo around the bacterial colony upon addition of HgCl2.
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2

Extracellular Enzyme Assays of Bacterial Strains

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Hemolysis assays on agar plates were conducted by picking a colony of each strain previously grown on TSA-1 and inoculating it on sheep blood agar plates (Oxoid) followed by growth at 25°C. For the phospholipase/lecithinase activity assay, 3 μl of overnight cultures in TSB-1 were spotted onto TSA-1 plates supplemented with 3% egg yolk extract (Oxoid), and results were evaluated after 24 h of culture at 25°C. Hydrolysis of lecithin by the phospholipase yields water-insoluble diglycerides that cause the appearance of an opaque precipitate. The gelatinase activity assay was carried out by spotting 3 μl of a TSB-1 overnight culture onto TSA-1 plates supplemented with 1% gelatin (Oxoid), and results were developed after 48 h of incubation at 25°C by covering the agar plate surface with a 12.5% (wt/vol) HgCl2 solution. Hydrolysis of gelatin by the gelatinase enzyme causes the appearance of a translucent halo around the bacterial colony upon addition of HgCl2.
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3

Evaluating Bacterial Virulence Factors

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Hemolysis and phospholipase activities were evaluated on TSA plates supplemented with 5% sheep blood agar (Thermo Scientific) and 3% egg yolk extract (Oxoid), respectively. To determine the impact of NaCl concentration in the phenotypical observation of these two activities, NaCl was added to a final concentration of 1% or 3% during plate preparation. RM-71 cultures were grown in TSB-1 and TSB-3 up to an OD600 of 1.2, and 2 µL of the suspension were spotted on each assay plate and incubated at 25°C for 24–48 hours. The hemolytic and phospholipase activities were calculated by dividing the halo diameter by the colony diameter. A Student’s t-test was used to determine statistical significance.
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