TITTLE: SELECTIVE AND DIFFERENTIAL MEDIA

AIM:
To characterize and describe the growth and colony appearance of distinct species of
bacteria inoculated on three different media, Eosin Methylene Blue agar, Mannitol
Salt Agar, and Blood agar.

OBJECTIVES
▪ To study the functional basis of different media
▪ To explain the results observed in this experiment
INTRODUCTION
Differential media is used to differentiate bacteria with morphological and
biochemical similarities. It has substances incorporated in it, enabling it to bring out
differing characteristics of bacteria and thus helping to distinguish between them
(Kumar, 2015). After incubation, different species of bacteria give characteristic
colony morphologies. The differential media used in this experiment was Eosin
Methylene Blue agar (EBM) which contains the dyes eosin and methylene blue
and it distinguishes strains based on lactose fermentation. As EMB contains
methylene blue that inhibits the growth of Gram-positive bacteria it is also a selective
medium. Another differential and selective media used is Mannitol salt agar (MSA)
and it contains 7.5% sodium chloride which helps select only those bacteria that
can tolerate high salt concentrations however Staphylococci species can tolerate
this salt concentration, but other pathogenic bacteria may not, the sugar mannitol
which is a differential ingredient in MSA. Organisms which can use mannitol as a
food source will produce acidic byproducts of fermentation that will lower the pH
of the media and it is this acidity that will cause the pH indicator, phenol red, to
turn yellow. Blood agar which is both a differential and selective medium as it can
help detect and differentiate hemolytic bacteria such as streptococcus species,
detect hemolysis by cytolytic toxins which are secreted by a strain of bacteria.
PROCEDURE
As per the manual
RESULTS

Bacteria Observation
MSA EBM BLOOD AGAR
Escherichia Coli No growth Green-metallic
(E.c) sheen colonies
Staphylococcus White colonies No growth
epidermis ( S.e)
Staphylococcus Yellow colonies No growth
aureus (S.a) with yellow zones
Microorganisms No growth No growth
from the streak
plate
Bacteria from the White colonies
throat

DISCUSSION
Escherichia Coli had no growth on MSA
Mannitol Salt Agar (MSA) is a type of medium that is both selective and
differential (Primm. & Siegesmund, 2017). As seen on the results, Escherichia Coli
does not grow on MSA because it is gram-negative and gram-negative are not
tolerant to salt (sodium chloride) which constitutes MSA. Staphylococcus aureus,
ferments mannitol and as a result turns the medium yellow as seen on the results.
Likewise, Staphylococcus epidermidis is salt-tolerant (halophilic) and will produce
colonies on MSA. Microorganisms from the streak plate fail to grow on MSA
because of their inability to tolerate high salt environment. EBM is a both
selective and differential media and it contains eosin and methylene blue dyes
which inhibit the growth of gram-positive bacteria and therefore select the
growth of gram-negative bacteria, it also contains lactose which differentiates
non-enteric bacteria based on their ability to ferment lactose. Only Escherichia
Coli grows on EBM because it ferments lactose and produce strong acids reducing
the pH of the EBM agar hence the formation of the green metallic sheen colonies
observed. White colonies form on the culture of bacteria from the throat on the
blood agar because it is normally nonselective and permits the growth of normal
throat flora as well as other potential pathogens
CONCLUSION
The practical was carried out successfully and different principles of different
growth media were studied and results were recorded.
REFERENCES
Bauman. R (2009) Microbiology with Diseases by Body System, 2nd ed. Pearson
Education, Inc.
Primm. P & Siegesmund. M (2017), Microbiology with diseases by body system 5th
ed Pearson Education, Inc.
Kumar.S (2015), Essentials of Microbiology, Jaypee Brothers Medical Publishers