Microbio Lab 6
Microbio Lab 6
Microbio Lab 6
USTMED ’07 Sec C – AsM; Photos provided by JV.N & MeaM. rhusiopathiae. Colonies are
smooth, small (less than
Amino acid hydrolysis reactions of Nocardia asteroids. All 1.0mm in diameter), and
amino acid reactions are negative on this identification plate: transparent. A greenish color
casein, tyrosine, xanthine and hypoxanthine. and/or α-hemolysis appears as
the colonies age.
Erysipelothrix rhusiopathiae on
tripe sugar iron (TSI) agar.
Erysipelothrix rhusiopathiae
demonstrates H2S production along
the stab line in a TSI agar slant within
48 hours of incubation at 35oC. This is
a helpful feature that separates this
organism from other gram-positive
bacilli.
Colonies of Nocardia asteroides on 5% sheep blood agar.
Colonies initially appear as powdery, and as they mature they
can develop a number of colors, ranging from a chalky-white to
yellow-orange, buff, red, purple, brown and black. Most
colonies have an earthy odor. Amino acid hydrolysis
reactions of
Streptomyces spp. All
reactions are positive on
this identification plate.
Unlike any Nocardia spp.,
Streptomyces spp.
hydrolyze casein,
tyrosine, xanthine, and hypoxanthine. Colonial morphology
resembles the chalky, dry white colonies of Nocardia spp.
Although Streptomyces spp. are urea-positive,
they are nitrate-negative, distinguishing the
genera from Nocardia spp.
Colonies of Nocardia asteroids growing on BAY late (x15).
Colonies are large, dry, bumpy, and heaped after 9 days
Molar tooth appearance of Actinomyces israelii
incubation at 27oC. This morphology has been referred to as
after incubation for 1 week.
glabrous orange form.
Gram stain Nocardia spp. (x1250). Nocardia species are Skin punch biopsy with
branching, beaded, filamentous gram-positive bacilli, positive acid fast stain of
approximately 1um in diameter. They can also appear as Mycobacterium leprae.
coccoid or coccobacillary forms. Care should be taken when
examining the slides because of the faint staining properties of
these microorganisms and other actinomycetes.
Colonies of M.
tuberculosis growing on
Middlebrook agar.
Cording characteristics of
M. tuberculosis.
Colonies of Bacteroides
fragilis on Bacteroides bile
esculin (BBE) agar. This
medium contains gentamicin to
Sputum smear stained with inhibit facultative gram-
Gram’s stain shows negative bacilli. Bile salts
neutrophils, amorphous inhibit most anaerobes other
debris, and filamentous, than those of the B. fragilis
beaded, branched gram- group, which grow as large,
positive bacilli (oil gray to black colonies due to
immersion) their hydrolysis of esculin in the medium.
Clostridium botulinum
Colonies of Bacteroides
fragilis on Brucella blood
agar. Bacteroides spp. as
well as other anaerobic
gram-negative bacilli grow
best on this nonselective
mediu. Agar must be
supplemented with hemin
and vitamin K1 to support Reverse CAMP test for
the growth of many species presumptive identification
of anaerobes. These colonies appear as circular, entire and of Clostridium perfringens. C. perfringens is streaked verticall
raised. and Streptococcus agalactiae is streaked horizontally in this
test. The hemolysis of the clostridium is synergistically
Colonies of Listeria enhanced by the hemolysin of the streptococcus in an
monocytogenes on 5% sheep arrowhead-shaped pattern.
blood agar. Colonies of
Listeria monocytogenes are Clostridium tetani on
small (less than 1mm in Brucella blood agar.
diameter), smooth, irregular, Growth on agar is
and translucent. The colonies characterized by
are surrounded by a very swarming, such that
characteristic narrow zone of beta hemolysis on 5% sheep blood growth will cover the
agar and may be confused with group B beta-hemolytic entire plate in a thin film
streptococci. Differentiating characteristics include Listeria’s within a few days.
narrow zone of beta hemolysis, positive catalase production,
and growth on bile and hydrolysis of esculin. Both species
hydrolyze hippurate.
Clostridium perfringens
on Brucella blood agar.
Colonies display the
Colonies of C. double-zone of beta
diphtheriae on 5% hemolysis typical of this
sheep blood agar. pecies. Colonies are large
Corynebacterium with peaked centers and
diphtheriae grow well on irregular edges after 48
5% sheep blood agar as hours incubation.
whitish, opaque colonies
Clostridium difficile and C.
perfringens on egg yolk agar.
The C. perfringens (upper half of
plate) produces abundant
lecithinase, while the C. difficile
(lower half) grows but shows no
reaction.
Gram stain of
Clostridium spp. (x1250).
Cells are parallel-sided,
long, thin, gram-variable,
and some show swollen
ends indicative of spore
formation.
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