Name: Velchez, Sheila Mae Date: 05/11/23

Section: BSMLS 2-B-10 Group #: 2

A. Discuss the basis of Biochemical Tests.

- Biochemical tests are laboratory techniques used to analyze and identify specific
chemical reactions and metabolic processes occurring in biological systems. These tests
are widely used in various fields such as clinical diagnostics, microbiology, biochemistry,
and genetics. They provide valuable information about the composition, function, and
characteristics of biological molecules, cells, and organisms.

The basis of biochemical tests lies in the fact that different substances exhibit distinct
chemical properties and reactions. By subjecting biological samples to specific test
reagents or conditions, scientists can observe and interpret the resulting changes or
reactions, which provide clues about the presence or absence of certain biochemical
compounds or activities.

Here are some common principles and basis for biochemical tests:

Enzymatic Reactions: Many biochemical tests rely on the detection of specific enzymes
or enzyme activities. Enzymes are proteins that catalyze chemical reactions in living
organisms. By introducing a substrate and observing the conversion of the substrate into
a product, the presence or absence of a particular enzyme can be determined. Examples
include the detection of specific enzymes in clinical blood tests, such as liver function
tests.

Chemical Reactions: Biochemical tests also exploit the chemical properties of

substances. For example, a color change or precipitation reaction may occur when a
specific chemical reagent reacts with a particular molecule or compound. These
reactions can indicate the presence or absence of specific substances. The Biuret test,
which detects the presence of proteins based on a color change, is an example of a
chemical reaction-based biochemical test.
Metabolic Pathways: Many biochemical tests are designed to assess the activity of
metabolic pathways in cells. These tests measure the production or consumption of
specific metabolites or the release of byproducts. For example, the glucose tolerance
test measures how effectively the body metabolizes glucose and can aid in diagnosing
diabetes.

Antibody-Antigen Interactions: Immunoassays are biochemical tests that rely on the

specific binding of antibodies to antigens. Antibodies are produced by the immune
system in response to foreign substances or specific target molecules. By using labeled
antibodies that recognize and bind to a specific antigen, scientists can detect and
quantify the presence of the antigen. These tests are widely used in medical diagnostics,
such as the detection of infectious agents or the measurement of hormone levels.

Molecular Techniques: With advancements in molecular biology, biochemical tests now

incorporate techniques like polymerase chain reaction (PCR), DNA sequencing, and gene
expression analysis. These techniques allow the detection, identification, and analysis of
specific nucleic acid sequences, genetic mutations, or gene expression patterns,
providing valuable information about genetic disorders, microbial identification, or gene
regulation.

Overall, biochemical tests are based on the principles of specific chemical reactions,
enzymatic activities, metabolic pathways, immune interactions, and molecular
techniques. These tests enable researchers and healthcare professionals to identify and
understand the biochemical characteristics of biological samples, contributing to
diagnosis, research, and the advancement of various scientific fields.
B. Enumerate and briefly discuss the different biochemical tests for Enterobacteriaceae
identification.

- Enterobacteriaceae is a family of gram-negative bacteria that includes several clinically

relevant genera such as Escherichia, Salmonella, Klebsiella, and Enterobacter.
Identification of Enterobacteriaceae is crucial for diagnosing and treating infections
caused by these bacteria. Various biochemical tests are commonly used to differentiate
and identify different members of the Enterobacteriaceae family. Here are some of the
key tests:

Catalase Test: This test determines the presence of the enzyme catalase, which breaks
down hydrogen peroxide into water and oxygen. Enterobacteriaceae bacteria are
catalase-positive, producing bubbles of oxygen when hydrogen peroxide is added.

Oxidase Test: The oxidase test is used to identify bacteria that produce the enzyme
cytochrome oxidase. Enterobacteriaceae bacteria are oxidase-negative, meaning they do
not produce this enzyme.

Triple Sugar Iron (TSI) Agar: TSI agar is a differential medium used to identify
Enterobacteriaceae based on their ability to ferment sugars and produce hydrogen
sulfide (H2S) gas. It contains three sugars (glucose, lactose, and sucrose) and an indicator
for acid production (phenol red) and H2S production (ferrous sulfate). The appearance of
yellow (acid production), black (H2S production), or red (no fermentation) can provide
clues for identification.

Citrate Utilization Test: This test determines the ability of bacteria to use citrate as a
sole carbon source. A positive result is indicated by a change in the color of the medium
from green to blue due to the production of alkaline byproducts.

Indole Production Test: This test detects the production of indole, a byproduct of
tryptophan metabolism. Bacteria that produce the enzyme tryptophanase can convert
tryptophan into indole. The addition of Kovac's reagent to a tryptone broth inoculated
with the bacteria produces a red color in the presence of indole.

Methyl Red and Voges-Proskauer (MR-VP) Tests: These tests are performed together to
differentiate between two different metabolic pathways. The Methyl Red test detects
the production of stable acids as a result of glucose fermentation, while the Voges-
Proskauer test detects the production of acetoin, a precursor to 2,3-butanediol, by
certain bacteria.
Urease Test: This test determines the ability of bacteria to produce the enzyme urease,
which hydrolyzes urea into ammonia and carbon dioxide. A positive test is indicated by a
color change due to the increase in pH caused by ammonia production.

These are just a few of the many biochemical tests used for the identification of
Enterobacteriaceae. In a clinical laboratory setting, a combination of these tests, along
with other tests like serotyping, antibiotic susceptibility testing, and molecular
techniques, is often used to accurately identify and differentiate the various members of
this bacterial family.