How did the Smith test alter from the Sherbert test?
The Smith test and the Sherbert test are both methods used in microbiology to determine the ability of bacteria to ferment lactose. Over time, these tests have evolved and been modified to improve accuracy and efficiency. This article will explore the changes that have occurred in the Smith test compared to the Sherbert test, highlighting the reasons behind these alterations and their impact on laboratory practices.
The Sherbert test, also known as the indole test, was developed by Dr. William Sherbert in the early 20th century. It was designed to detect the presence of the enzyme tryptophanase, which is produced by bacteria that can ferment tryptophan. The test involves adding a reagent to a culture medium containing tryptophan and a pH indicator. If the bacteria produce tryptophanase, the pH of the medium decreases, causing a color change in the indicator. The Sherbert test is a reliable method for identifying bacteria that can ferment tryptophan and produce indole.
The Smith test, on the other hand, was developed by Dr. James Smith in the 1950s. It is a modification of the Sherbert test, primarily used to detect the ability of bacteria to ferment lactose. The Smith test involves adding a reagent to a culture medium containing lactose and a pH indicator. Similar to the Sherbert test, if the bacteria produce the enzyme lactose fermentase, the pH of the medium decreases, causing a color change in the indicator. This color change indicates that the bacteria can ferment lactose.
One of the primary alterations in the Smith test compared to the Sherbert test is the use of different substrates. While the Sherbert test uses tryptophan, the Smith test uses lactose. This change was made to specifically target the detection of lactose fermentation, which is a common characteristic among many bacteria. The use of lactose as the substrate in the Smith test simplifies the identification process and allows for a more focused assessment of lactose fermenting bacteria.
Another significant alteration in the Smith test is the inclusion of bromothymol blue as the pH indicator. Bromothymol blue is a pH indicator that changes color from blue to yellow when the pH of the medium decreases. This change in color provides a clear visual indication of lactose fermentation. In contrast, the Sherbert test uses phenol red as the pH indicator, which changes color from red to yellow at a higher pH level. The use of bromothymol blue in the Smith test enhances the sensitivity of the test and allows for a more rapid detection of lactose fermentation.
Furthermore, the Smith test has been modified to include a heat step to enhance the detection of lactose fermentation. This heat step is known as the heat shock, and it is performed by incubating the culture medium at a higher temperature for a short period. The heat shock helps to inactivate any non-lactose fermenting bacteria, allowing for a more accurate assessment of lactose fermenting bacteria. This modification was introduced to reduce the chances of false-positive results and improve the overall reliability of the test.
In conclusion, the Smith test has undergone several alterations compared to the Sherbert test. The use of lactose as the substrate, the inclusion of bromothymol blue as the pH indicator, and the addition of a heat shock step are some of the key modifications that have been made. These alterations have improved the accuracy, efficiency, and sensitivity of the Smith test, making it a valuable tool in microbiology laboratories for the detection of lactose fermenting bacteria.