DISSCUSSION

1. What are the different characteristics of non-Enterobacteriaciae versus Enterobacteriaciae

group? (8 marks)
N NON - ENTEROBACTERIACIAE
O ENTEROBACTERIACIAE
1 Not part of normal flora Usually part of normal flora
2 Obligate aerobes Facultative anaerobes
3 Mostly oxidase positive Mostly oxidase negative
4 Do not use carbohydrates as a source Use carbohydrate as aa source of energy
of energy

2. Describe the mode of action of antibiotics against bacteria. (6 marks)

I. Cell Wall Synthesis Inhibitors:
Some antibiotics like beta-lactams, cephalosporins and glycopeptides inhibits the
ability of microorganism to synthesize the cell the wall which mainly contains murein
and peptidoglycan.
II. Protein Synthesis Inhibitors:
 Some antibiotics like Streptomycine, Erythromycine, tetracyclines, chloramphenicol,
lincomycin, kanamycin disulphate salts, aminoglycosides and macrolides interfere in
the formation of protein synthesis of microbes
III. Cell Membrane Inhibitors:
Antibiotics like polymyxins interrupt the integrity and structure of cell membranes,
hence killing them. These types of antibiotics are generally effective on gram –ve
bacteria as they have a definite cell membrane
IV. Effect on Nucleic Acids: 
The whole existence and life of any living system depends on DNA and RNA present
in every living cell. Some antibiotics like etopocide, rifampcin, quinolones and
rifamycins bonded with those proteins which are essential for the processing of these
nucleic acids. Hence these antibiotics block the synthesis of nucleic acid and therefore
affect the growth of the living cells.
V. Competitive Inhibitors: 
Some antibiotics competitively inhibit the important metabolic pathways happen
inside the bacterial cell. These antibiotics resemble a microbial substrate and compete
with that substrate for the limited microbial enzyme. The antibiotic ties up the enzyme
and blocks a step in metabolism. They are also called as anti-metabolites or growth
factor analogs. For example; Sulfonamides like Gantrisin and Trimethoprim
3. Give FOUR (4) the names of other media that commonly used to classify
Enterobacteriaciae group in the hospital and state it principles. (6 marks)
N MEDIA
O
1 MacConkey Agar
2 TCBS agar
PRINCIPLE
MacConkey agar is a selective and differential
culture media commonly used for the isolation of
enteric Gram-negative bacteria. It is based on the
bile salt-neutral red-lactose agar of MacConkey.
Crystal violet and bile salts are incorporated in
MacConkey agar to prevent the growth of Gram-
positive bacteria and fastidious Gram-negative
bacteria, such as Neisseria and Pasteurella.
Gram-negative enteric bacteria can tolerate bile
salts because of their bile-resistant outer
membrane. Gram-negative enteric bacteria that
grow on MacConkey agar are differentiated by
their ability to ferment lactose. If the lactose is
fermented by the bacteria, the production of the
acid drops the pH of the media. The drop in pH
is indicated by the change of neutral red indictor
to pink (Neutral read appears pink at pH’s below
6.8).
Strongly lactose fermenting bacteria produce
sufficient acid which causes precipitation of the
bile salts around the growth. It appears as a pink
halo surrounding individual colonies or areas of
confluent growth. Pink halo in not seen around
the colonies of weaker lactose fermenting
bacteria.Gram-negative bacteria that grow on
MacConkey agar but do not ferment lactose
appear colorless on the medium and the agar
surrounding the bacteria remains relatively
transparent.
TCBS Agar is used for the selective isolation of
Vibrio cholerae and other enteropathogenic
vibrios. Thiosulfate and sodium citrate, as well
as the alkalinity of the medium, considerably
inhibit the growth of Enterobacteria. Ox bile and
sodium cholate slow the growth of enterococci
and inhibit the development of Gram-positive
bacteria. The acidification of the medium
resulting from the fermentation of sucrose by
Vibrio makes bromthymol blue turn yellow.
Bromthymol Blue and Thymol Blue are pH
indicators. Using thiosulfate as a sulfur source,
the production of hydrogen sulfide is visualized
in the presence of ferric citrate. Yeast extract and
 peptone provides the nitrogen, vitamins, and
amino acids in TCBS Agar. Sodium chloride
provide optimum growth and metabolic activity
of halophilic Vibrio spp. Agar is a Solidifying
agent.

An increased pH is used to enhance growth of

Vibrio cholerae, because this organism is
sensitive to acid environments.

3 Salmonella-Shigella (SS) agar Salmonella-Shigella (SS) agar is a selective and

differential medium . It is used for the isolation,
cultivation and differentiation of gram-negative
enteric microorganisms isolated from both
clinical and non-clinical specimens such as from
feces, urine, and suspected food items (fresh and
canned foods). This medium is not
recommended for the primary isolation of
Shigella as some of Shigella strains may not
grow on SS agar due to relatively high level of
selectivity. The presence of bile salts mixture
and dyes (brilliant green) inhibits the growth of
gram-positive species to a varying degree.
Differentiation of enteric organisms is achieved
by the incorporation of lactose in the medium.
Organisms which ferment lactose produce acid
which, in the presence of the neutral red
indicator, results in the formation of red/pink
colonies. Lactose non-fermenters form colorless
colonies. The latter group contains the majority
of the intestinal pathogens, including Salmonella
and Shigella.The sodium thiosulfate and ferric
citrate enable the detection of hydrogen sulfide
production as evidenced by colonies with black
centers.

4 CLED Agar CLED Agar is an abbreviation for Cystine

Lactose Electrolyte-Deficient Agar. It is a type
of differential medium recommended for
diagnostic urinary bacteriology. The medium
supports the growth of all urinary potential
pathogens and provides distinct colony
morphology. CLED Agar also supports the
growth of a number of contaminants such as
diphtheroids, lactobacilli, and micrococci. It is
electrolyte deficient to prevent the swarming of
Proteus species. The nutrients in BD CLED Agar
are supplied by the gelatin and casein peptones,
and beef extract. Lactose is included to provide
an energy source for organisms capable of
utilizing it by a
fermentative mechanism. Bromthymol blue is
used as a pH indicator to differentiate lactose
fermenters from lactose-nonfermenters.
Organisms which ferment lactose will lower the
pH and
change the color of the medium from green to
yellow. The cystine permits the growth of "dwarf
colony" coliforms.3 Electrolyte sources are
reduced in order to minimize the swarming of
Proteus
species. Thus, the medium allows quantitative
determination of urinary pathogens including
Proteus when calibated loops are used for
inoculation