Before agar, microbiologists had experimented with other foodstuffs as microbial media. They turned to substances rich in the starches, proteins, sugars, fats, and minerals that organisms need for growth, testing with broths, bread, potatoes, polenta, egg whites, coagulated blood serums, and gelatine. However, none worked particularly well: all were easily broken down by heat and microbial enzymes, and their surface, once colonized, became mushy and unsuitable for isolating microbes.
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This was not the first time that microbiologists experienced problems with agar. A phenomenon called “The Great Plate Count Anomaly” baffled researchers in the early 20th century when they observed that the number of cells seen under a microscope didn’t match the actual number of colonies growing on an agar plate. Investigating this discrepancy, researchers found agar itself to be the culprit: when nutrient broths are heated with agar during boiling, harmful byproducts (hydroperoxide) can form due to the reaction of agar with phosphate minerals contained in the media. Researchers can avoid this by autoclaving agar separately from the nutrient broth, or by reducing the amount of agar used.，这一点在safew官方版本下载中也有详细论述
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