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Nitrate reduction test • 3 possibilities • Nitrate – nitrate reductase – nitrite • Nitrate – nitrate reductase – nitrite – Nitrite reductase – nitrogen gas • Neither enzymes were produced by the bacterium – bacterium cannot use nitrate. • Some bacteria make only nitrate reductase • Some bacteria make both nitrate reductase, nitrite reductase • Some bacteria neither of the enzymes • Nitrite indicators are added to each tube. • Medium becomes red – nitrite in the medium • Bacterium produced nitrate reductase only. • Add a pinch of zinc to those tubes that did not become red • Medium becomes red after adding zinc • Zinc converted nitrate in the medium to nitrite – chemical reduction, not bacterial reduction. • Bacteria did not use the nitrate. Therefore zinc was able to use it – convert it. • Bacterium produced neither of the enzymes • No red color after adding zinc - bacterial reduction • Bacteria made both nitrate reductase and nitrate reductase • Nitrate – nitrite – nitrogen gas Urea hydrolysis • Urea – urease – ammonium ions • Change the pH indicator to fuchsia Litmus milk • • • • • • Lactose – milk sugar Casein – milk protein Litmus – pH indicator Acidic – pink Basic – blue Neutral - lavender Streptococcus lactis • • • • • • Lactose – acids Acids then coagulate the milk protein Milk becomes a solid E. coli Not a strong acid producer as S. lactis Enough to change litmus to pink, but not enough to coagulate the protein Pseudomonas aeruginosa • Positive for peptonization ( protein hydrolysis). • Casein – amino acids • Milk becomes a brownish solution