MODULE 5 IN

CLINICAL BACTERIOLOGY

MLS 223

SCHOOL OF NATURAL SCIENCES

Department of Medical Laboratory Science

Prepared by:
Kathyren C. Estimada, RMT, MSMT
Arlene A. Mangiduyos, RMT

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 MODULE 5
GRAM-POSITIVE BACILLI

Describe each bacterium/related group of gram-positive bacilli as to:

a. general characteristics (morphology and physiology)
b. epidemiology (habitat and transmission)
c. pathogenesis and clinical manifestations
d. laboratory diagnosis
e. prevention and control

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 MODULE 5
GRAM-POSITIVE BACILLI

Unit 1: Non-spore-forming Gram-positive Bacilli

Unit Learning Outcome:

Describe the medically-significant non-spore-forming, gram-positive bacilli as to their

general characteristics, epidemiology, pathogenesis and clinical manifestations,
diagnosis, prevention and control.

Engage

In this module unit you shall encounter a large, diverse group of bacteria which consists
of those gram-positive non-spore-forming bacilli that are encountered in clinical
microbiology laboratory in varying frequencies.

Explore

The gram-positive non–spore-forming bacilli are a mixed group of genera subdivided

on the basis of morphology and staining characteristics. Bacteria that belong to the
gram-positive non–spore-forming bacilli group, includes the genera Corynebacterium,
Listeria, Erysipelothrix, and Lactobacillus.

A wide range of clinical conditions result from infection with these organisms. Although
several of these organisms are frequently isolated in the clinical laboratory, they are
typically considered contaminants or commensals (e.g., Corynebacterium and
Lactobacillus). Several of these organisms are rarely encountered but cause significant
disease (Listeria, Erysipelothrix, and Corynebacterium diphtheriae).

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 Explain

Corynebacterium diphtheriae

Common name: Klebs-Loeffler's bacillus

T he bacterium was first observed in diphtheritic membranes by Edwin Klebs in 1883 and cultivated by Friedrich Löffler in 1884.
T

GENERAL CHARACTERISTICS
• Gram-positive bacilli
• Pleomorphic, possessing irregular swellings at one end that give them the club-
shaped” appearance. The term diphtheroid, meaning “diphtheria-like,” is
sometimes used in reference to this Gram staining morphology.
• Arranged in pairs of cells at angles to one another (X, Y V, or L configuration, or
“Chinese letters”); or single cells that tend to lie parallel to each other
(“palisades”)
• Metachromatic granules (called volutin granules or Babe-Ernst granules) are
irregularly distributed in the cytoplasm (often near the poles) that give the rod
a beaded appearance in stained smear.
• Nonmotile
• Non-spore-forming
• Aerobic/Facultative anaerobic
• Catalase(+)
• Oxidase (+)

HABITAT AND TRANSMISSION

• Habitat is the human nasopharynx.
• Transmission is through direct contact with respiratory secretions or exudates
from skin lesions.

PATHOGENESIS AND CLINICAL MANIFESTATIONS

Diphtheria is an acute disease caused by toxigenic (toxin-producing) strains of C.

diphtheriae. The incubation period for diphtheria is 2 to 5 days, with a range of 1 to 10
days.

For clinical purposes, it is convenient to classify diphtheria by anatomic site:

a. Respiratory diphtheria (pharyngeal, tonsillar, laryngeal, nasal)

• The most common sites of diphtheria infection are the
pharynx and the tonsils.
• Characterized by gradual onset of pharyngitis; early
symptoms include sore throat with low-grade fever and
dyspnea.
• Localized manifestations:
- Pseudomembrane forms in the throat (i.e., gray-white Respiratory diphtheria
patches composed of fibrin, necrotic host cells and results in pseudomembrane
bacteria) over the pharynx, tonsils, uvula, and palate. formation.
The membrane is firmly adherent to the tissue, and

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 forcible attempts to remove it cause bleeding. Extension of membrane
formation into the larynx, or anterior nasal may result in respiratory
obstruction (suffocation), coma, and death.
- In severe disease, marked edema of the
submandibular areas and the anterior neck along
with lymphadenopathy giving a characteristic "bull
neck" appearance is usually present.
• Systemic manifestations:
- Major complications include myocarditis and
p o l y n e u r i t i s , a n d a l s o n e p h r i t i s a n d Enlarged regional lymph
thrombocytopenia nodes in the neck with marked
i. Paralysis of eye muscles, limbs, and the edema, referred as “bull
diaphragm can occur after the fifth week. neck”, is present in severe
disease.
Secondary pneumonia and respiratory
failure may result from diaphragmatic
paralysis.
ii. Damage to the heart causes heart failure, which is the most
common cause of mortality in diphtheria.
- Death occurs in 5-10%.

b. Cutaneous diphtheria (wound diphtheria)

• Presents as infected skin lesions which lack a
characteristic appearance; a membrane forms on
the infected wound that fails to heal.
• May be associated with non-toxigenic strains of C.
diphtheriae
• Appears to result less frequently in systemic Cutaneous diphtheria is often
complications seen with pseudomembrane
forming over on an infected
wound that fails to heal.

Corynebacterium ulcerans and Corynebacterium pseudotuberculosis are closely related to C. diphtheriae

and may carry the diphtheria tox gene. Both are zoonotic organisms. Whereas the toxigenic C. ulcerans can
cause disease similar to clinical diphtheria, C. pseudotuberculosis rarely causes disease in humans.

VIRULENCE FACTORS

Diphtheria exotoxin (Diphtherotoxin)

• Produced by lysogenized strains of C. diphtheriae infected by β-prophages
(corynebacteriophages) that carry the tox gene; all strains produce the same
antigenic type of toxin.
• Absorption of toxins through the mucous membranes and into the blood
circulation causes local tissue destruction damage to the peripheral nervous
system, heart, and other organs of the body.
• Diphtheria toxin consists of two polypeptide fragments:
a. Fragment A interacts metabolically with factors in the cytoplasm and
stops protein synthesis by inhibiting polypeptide chain elongation. It is
assumed that the abrupt arrest of protein synthesis is responsible for the
necrotizing and neurotoxic effects of diphtheria toxin.
b. Fragment B binds to and facilitates the entry of the toxin into the
cytoplasm of the cells of the heart and nervous system through
receptor-mediated endocytosis.

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PREVENTION AND CONTROL
• Diphtheria can be easily prevented by a series of vaccinations with diphtheria toxoid,
a modified diphtheria toxin that induces protective antitoxin antibodies. Toxin-
producing C. diphtheriae is grown in liquid media and the toxin is converted to the
inactive toxoid by treatment with formalin. It is usually given as part of a vaccine
against tetanus and pertussis called the DPT (diphtheria, pertussis, tetanus) vaccine.
Three vaccinations are recommended, starting at 6-8 weeks of age followed by at 15
months and again at school age. DPT-containing multi-antigen vaccines (with Hep B,
Hib, or IPV --- respectively, Hepatitis B vaccine, Haemophilus influenzae type B
vaccine, or inactivated poliovirus vaccine) are increasingly being used.
• Persons with suspected respiratory diphtheria should be promptly given diphtheria
antitoxin, produced in horses, in adequate dosage, without waiting for laboratory
confirmation. Diphtheria antitoxin will neutralize circulating toxin and prevent
progression of disease, but does not neutralize toxin that is already fixed to tissues,
• Diphtheria infections are also managed by chemotherapy, i.e., patients are treated
with antibiotics. The disease is usually no longer contagious 48 hours after antibiotics
have been given. Antibiotic therapy should not be relied alone. This must be done in
addition to antitoxin therapy,

Schick test is performed to determine the following:

• susceptibility (due to lack of antitoxins), or immunity (due to presence of circulating
antitoxins) of an individual against diphtheria.
• hypersensitivity to diphtheria toxin or other proteins of the diphtheria cell.

This serves as a basis whether a person may receive vaccination against diphtheria
or not. Individuals who are susceptible to diphtheria are to be vaccinated, and if they
are hypersensitive, they should be vaccinated with caution --- the doses of the
vaccine shall be reduced with increased number of injections. It should also be Diphtheria toxin
performed first before antitoxin therapy since antitoxin is normally of horse origin,
which may cause hypersensitivity.

The procedure involves injection of 0.1 mL of diphtheria toxin on the test arm (TA)
and 0.1 mL of diphtheria toxoid (vaccine preparation or heat-inactivated toxin ---
whose toxicity is destroyed but retains capacity to elicit allergic reactions) on the
control arm (CA)
• The injection site (TA and CA) is inspected daily up to the 6th day for erythema,
induration or necrosis which consists of a positive reaction.
• Results interpretation are as follows:
(1) POSITIVE REACTION:
Diphtheria toxoid
TA – reaction persists until the 6th day
CA – no reaction
*Susceptible; NOT hypersensitive
(2) NEGATIVE REACTION:
TA – no reaction
CA – no reaction
*Immuned; Not hypersensitive
(3) COMBINED REACTION:
TA – reaction persists until the 6th day
CA – reaction peaks at about 48 hours and subsides by
Positive reaction in Schick test.
day 5
*Susceptible and hypersensitive
(4) PSEUDOREACTION:
TA – reaction subsides by day 5
CA – reaction subsides by day 5
*Immuned and hypersensitive

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LABORATORY DIAGNOSIS

Specimens

• Swabs from the oropharynx

If a pseudomembrane is present, swab specimens from beneath the
membrane should be collected.

• Swabs from the nasopharynx, or from cutaneous lesions.

A. Microscopy

• Gram staining: Pleomorphic gram-positive rods

that occur in angular arrangements (commonly
referred to as Chinese letters or palisades) and
whose ends may be swollen, producing a club
shape.

• Loeffler's alkaline methylene blue (LAMB), e.g.,

Albert's staining: Pleomorphic beaded rods,
reddish purple metachromatic granules or bars C. diphtheriae stained with LAMB.
of polyphosphates are apparent.

C. ulcerans and C. pseudotuberculosis resemble C. diphtheriae, producing distinctive, club-

shaped, diphtheroidal cells containing metachromatic granules.

B. Cultural method

• Culture media

1. 5% sheep BAM, Columbia CNA agar

- Used to screen and rule out group A β-hemolytic streptococci
from throat specimen.

2. Media containing cystine and potassium tellurite - selective and

differential for Corynebacterium species.

a. Tinsdale agar (TIN) - AKA Cystine-sodium thiosulfate-tellurite

medium

- High concentration of potassium

tellurite is inhibitory to most upper
respiratory tract normal flora
(other than Corynebacterium
species) and majority of gram-
negative bacteria; organisms
c a p a b l e o f g ro w i n g on t h e
medium are differentiated based
on the tellurite reductase activity,
resulting in the reduction to Tinsdale agar.
tellurium, thus will grow as black
colonies.

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 - Bovine serum and horse serum provide essential growth factors.

- Sodium thiosulfate provide sulfur for H2S production; and cystine

detects cystinase activity producing brown halos around the
colonies.

b. Cystine-tellurite blood agar (CTBA)

- A heart infusion agar supplemented

with 5% rabbit blood, tellurite, and L-
cystine.

- A selective and differential medium

for Corynebacterium species similar Cystine-tellurite-blood agar.
to TIN.

3. Loeffler's serum medium

- Contains eggs and beef serum, rather

than agar, to coagulate to produce a
solid medium.

- Stimulates the growth of distinctive,

club-shaped, diphtheroidal cells of C. Loeffler's serum medium
diphtheriae and the production of
metachromatic granules in the cells.

- "Poached egg" colonies show no characteristic differential

features to distinguish from other gram-positive bacilli.

4. Pai medium

- Contains coagulated egg in distilled water and glycerin.

• Inoculation and incubation

- Specimens should be inoculated to a blood agar and a selective

medium such as a tellurite plate (eg, CTBA or TIN)

- Incubate at either in ambient air or in 5% to 10% CO2 for 24 - 48 h.

• Colonial characterization

- C. diphtheriae may appear as four distinct colony types (biotypes)

designated gravis, mitis, intermedius, and belfanti.

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 These biotypes also differ slightly in Gram’s stain morphology,
certain biochemical reactions, and historically, in the severity of the
disease processes they produce.

- On BAM:

C. diphtheriae produce colonies

ranging from small, gray, and
translucent (biotype intermedius) to
medium, white, and opaque
(biotypes mitis, belfanti, and gravis);
C. diphtheriae biotype mitis may be β-
hemolytic. Colonies of C. diphtheriae on
BAM.

- On CTBA:

Colonies of C. diphtheriae appear

black or gray.

Colonies of C. diphtheriae on
CTBA.

Growth on CTBA Starch &

Gram Stain NO3
Biotypes Glycogen
Morphology reduction
Colonial Morphology Hemolysis Hydrolysis

Large (2-4 mm),

flatter, dark gray
with radial
Short,
striations and
C. diphtheriae coccoid, or
irregular edges. - + +
biotype gravis pyriform.
“DAISY HEAD"
colonies.

Small (0.5 mm),

sometime
Highly
pinpoint,
C. diphtheriae pleomorphic,
flat, and gray.
biotype - from very long - +
intermedius to very short
“FROG’S EGGS"
rods.
colonies.

Medium-sized
C. diphtheriae (1-2 mm), convex,
+
biotype mitis very black with Long,
May be
regular edges. pleomorphic,
weakly β- -
rigid club-
hemolytic.
“COOLIE HAT" shaped rods.
C. diphtheriae
colonies. -
biotype belfanti

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 - On TIN:

All four biotypes of C. diphtheriae grow

as black colonies surrounded by dark
brown halos.

Colonies of C. diphtheriae on TIN.

Colonies of C. diphtheriae on TIN.

C. ulcerans and C. pseudotuberculosis resemble C. diphtheriae, producing black colonies

that are surrounded by brown halos on TIN.

Other diphtheroids, S. aureus and some streptococci which can

also grow in the presence of tellurite will develop black
colonies due to tellurite reduction although brown halos around
the colonies are NOT present.

Acid production from

NO3 Reverse
Urease Gelatinase reduction Lipophilic
Starch or
Glucose Maltose Sucrose Lactose Glycogen CAMP*

- + -
C. except except except
+ + - - - V- -
diphtheriae biotype biotype biotype
gravis belfanti intermedius

C. ulcerans + + - - + + + - + -

C. pseudo-
+ + V - - + - V + -
tuberculosis

* The reverse CAMP reaction is based on the

production by the organism of phospholipase C or D
that inhibits the staphylococcal β-hemolysin.
An arrowhead zone of NO hemolysis is formed at the
junction of the organism being tested with the
staphylococci.

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D. Toxigenicity test

• Once an organism is biochemically identified as a possible C. diphtheriae,

the isolate must be tested for the ability to produce diphtheria toxin.

• Elek Test
- For demonstration of toxin production by C. diphtheriae.

A filter paper strip impregnated with diphtheria antitoxin is

buried just beneath the surface of a rabbit serum agar plate
before the agar hardens. Strains to be tested and known
positive and negative toxigenic strains are streaked on the
agar’s surface in a line across the plate and at a right angle to
the antitoxin paper strip. After 24 hours of incubation at 37°C,
the plates are examined with transmitted light for the presence
of fine precipitin lines at a 45-degree angle to the streaks. The
presence of precipitin lines indicates that the strain produced
toxin that reacted with the homologous antitoxin. Line 1 is the
negative control. Line 2 is the positive control. Line 3 is a test
organism that is a nontoxigenic strain. Line 4 is a test organism
that is a toxigenic strain.

(1)

(2)

? Quick Quiz!
(3)
Which strain (1, 2, or 3) of
C. diphtheriae is toxigenic?

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 Elaborate

Other Corynebacterium species

Corynebacterium jeikeium is most commonly isolated from acutely ill or

immunocompromised patients. It produces infections, including bacteremia that have
a high mortality rate and is resistant to many commonly used antimicrobial drugs.
Corynebacterium pseudodiphtheriticum is a normal inhabitant of the human
nasopharynx, can colonize natural and artificial heart valves. It has been associated
with respiratory tract infections.
Corynebacterium xerosis normally lives in the eye, skin, and mucous membranes and is
an occasional opportunist in eye and postoperative
infections.
Corynebacterium minutissimum is the causative agent of
erythrasma, a superficial skin infection characterized by
small, brown-red macular areas, commonly of the axillary (a) (b)
and inguinal skin, and also between the toes and finger, Erythrasma of the axillary and inguinal skin.
axilla. It gives a coral (brick red) fluorescence when
exposed to Wood’s light (long-wave UV radiation, also
called black light) due to porphyrin.
Corynebacterium striatum has been associated with
hospital-acquired respiratory tract and other infections.
Erythrasma under Wood's light shows brick red
Corynebacterium urealyticum is a slowly growing species fluorescence.
that is multiply resistant to antibiotics. It has been associated with acute or chronic
encrusted urinary tract infections manifested by alkaline urine pH and crystal formation.
Corynebacterium amycolatum is most commonly isolated species in the clinical
laboratory and may represent a skin contaminant or a significant agent of infection

Acid production from NO3 Reverse Lipophilic

Urease Gelatinase
reduction
Glucose Maltose Sucrose Lactose CAMP **

C. jeikeium + V - - - - - - +
C. pseudodiph-
- - - - + - + - -
theriticum
C. xerosis + + + - - - V - -
C. minutissimum + + + - - - - - -
C. striatum + - V+ - - - + V -
C. urealyticum - - - - + rapid - - - +
C. amycolatum + V+ V+ - V- - V+ - -

** Separation of lipophilic and nonlipophilic species can be determined by comparing growth on sheep blood agar and sheep blood agar with
1% Tween 80 or growth in brain-heart infusion broth with and without 1 drop of Tween 80 or rabbit serum.

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 Listeria monocytogenes

Named after Joseph Lister, the English surgeon who pioneered antiseptic surgery; the species name is derived from its effect on
monocytes, i.e., marked monocytosis in some animals.

CHARACTERISTICS
• Small, gram-positive bacilli (coccobacilli), resembling corynebacteria in
Chinese letter configuration
• Nonmotile at 37oC; motile at 25oC (22–28°C)
• Non-spore-forming
• Nonencapsulated
• Facultative anaerobic
• Catalase (+)
• Oxidase (+)
• Is capable of growing and surviving over a wide range of environmental
conditions: refrigerator temperatures (4°C), low pH, and high salt

HABITAT AND TRANSMISSION

• Reservoir is widespread. The organisms can be isolated from the environment
--- soil, water, and vegetation. They are part of the fecal flora of a wide
variety of animals. They are transient constituents of the human intestinal flora
and may be excreted in the feces by 1-10% of healthy humans.
• Human infections in adults or children are
usually food-borne, associated with ingestion
of contaminated milk, cheeses, ice cream,
raw vegetables, poultry, and meat.
• Vertical transmission can occur prenatally ---
across the placenta, or postnatally --- by
contact during delivery.

PATHOGENESIS AND CLINICAL MANIFESTATIONS

• Disease is known as listeriosis.
• Are facultative intracellular pathogens, within
phagocytes and non-phagocytic cells; the
organisms can penetrate the epithelial cells of
the gastrointestinal (GI) tract and grow within
hepatic and splenic macrophages; from
there, the organism can spread to the CNS or
the pregnant uterus.
• Clinical manifestations of listeriosis differ The principal routes of infection of listeriosis.
among the high-risk groups.
1. In normal adults, it is often a mild or subclinical infection with non specific
symptoms of fever, diarrhea, and sore throat.
2. In children and immunocompromised adults, esp. renal transplant
patients and cancer patients, and the elderly, listeriosis usually presents as
meningitis and septicemia.
3. In pregnant women, most common in the third trimester, listeriosis presents
as mild flu-like illness without meningitis, Bacteremia occurs concomitantly,
during which time the uterine contents are infected. Progression to
amnionitis may lead to abortion, stillbirth, or delivery of an acutely ill
infant.

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 4. In neonates, listeriosis may have an early or late onset.
a. Early-onset syndrome (granulomatosis infantiseptica) is the result of
infection in utero and is a disseminated form of the disease
characterized by neonatal sepsis, pustular lesions, and granulomas
containing L monocytogenes in multiple organs. Death may occur
before or after delivery.
b. The late-onset syndrome causes the development of meningitis
between birth and the third week of life; it is often caused by serotype
4b and has a significant mortality rate.

VIRULENCE FACTORS
• Adhesin proteins (Ami, Fbp A, and flagellin) which facilitate bacterial binding
to the host cells.
• Internalins (A and B) are cell wall surface proteins that interact with E-cadherin,
a receptor on epithelial cells, promoting phagocytosis of L. monocytogenes
cells into the epithelial cells.
• Listeriolysin O is an enzyme produced by the bacterium at low pH. It lyses the
phagolysosome membrane allowing the listeriae to escape into the cytoplasm
of the epithelial cell.
• Act A, another listerial surface protein, induces host cell actin polymerization,
which propels the listeriae to the cell membrane of the host’s epithelial cells
and permits it to move from cell to cell without being exposed to antibodies,
complement, or polymorphonuclear cells.

PREVENTION AND CONTROL

• Prevention can be improved by:
- avoidance of unpasteurized dairy products
- adequate pasteurization temperatures and thorough cooking foods
that are suspected of being contaminated with animal manure or
sewage
• Unlike diphtheria, there is no vaccine available to prevent listeriosis.
• Cold storage is not an effective control measure because the microbe can
grow at most refrigeration temperature.

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LABORATORY DIAGNOSIS

Specimens: : CSF, blood, amniotic fluid/placenta

1. Microscopy:
- Gram staining: Short, gram-positive bacilli
(coccobacilli)may occur intracellularly or
extracellularly, at times, may occur in pairs and
be mistaken for pneumococci; it tends to
appear gram-negative when overdecolorized.
- In wet mounts observed at 25oC, it exhibits
“tumbling" or "head over heels” motility.

2. Culture: L. monocytogenes in gram-

- Cold enrichment technique: Recovery of the stained CSF sediment.
organism from clinical samples is enhanced by
inoculation in broth medium and incubation at
4°C for 4 hours prior to incubation on primary culture plates.
- Requires 5% to 10% CO2 for growth.
- Media:
a. 5% sheep BAM
- L. monocytogenes has a narrow zone of beta-hemolysis on
sheep BAM.
b. McBride Agar
- McBride Agar can be used as a plating medium with or without
supplementation of blood.
- Partial selectivity is provided by lithium chloride, glycine and
phenylethanol, which aid in suppressing both G+ and G-
bacteria other than Listeria.
- L. monocytogenes develops into small, blue-green colonies,
with small zone of hemolysis around and under colonies.

L. monocytogenes
on sheep BAM.

L. monocytogenes
on McBride agar.

3. Motility test: In semi-solid medium, the identification

can be made by observing for a thin, umbrella-like
growth from the stab line.

Growth of L. monocytogenes
stabbed into a semi-solid medium.

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 4. Anton Test: This is a pathogenicity test that differentiates pathogenic from
non-pathogenic strains of L. moncytogenes. It Involves instillation of a 24hr
broth culture of L. monocytogenes in the conjunctiva of a lab animal.
Development of purulent conjunctivitis within 24 to 36 hours constitutes a
positive result.

(a) and (b) show a positive Anton test.

Erysipelothrix rhusiopathiae

• Gram-positive, nonmotile bacilli that are arranged singly, in short chains, or in long
non-branching filaments. It sometimes look gram-negative since it decolorizes easily.
• Forms small, transparent, and glistening colonies that may be alpha-hemolytic or
gamma-hemolytic on blood agar medium.
• Catalase(-), oxidase (-).
• Exhibits a “test tube brush-like” or "pipe cleaner" growth in gelatin stab culture

A. Gram reaction and morphology of

E. rhusiopathiae.
B. E. rhusiopathiae colonies on BAM
C. Growth of E. rhusiopathiae in a
semi-solid medium A B C
• E. rhusiopathiae is distributed in land and sea animals worldwide, including a variety
of vertebrates and invertebrates. It causes disease in domestic swine, turkeys, ducks,
and sheep. The most important impact is in swine, in which it causes erysipelas.
• Humans, acquire E. rhusiopathiae infection by direct
inoculation from animals or animal products. Persons at
greatest risk are fishermen, fish handlers, abattoir workers,
butchers, and others who have contact with animal products.
• The most common E. rhusiopathiae infection in humans is
erysipeloid, a nodular type of cellulitis. It usually occurs on the
fingers by direct inoculation at the site of a cut or abrasion
(and has been called “seal finger” and “whale finger”). Erysipeloid on the hand of an
• Additional clinical forms of infection (both rare) are a diffuse animal handler.
cutaneous form and bacteremia with or without endocarditis.
Septic arthritis has also been reported.

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• Prevention and control:
- Swine erysipelas can be prevented by vaccinating pigs, but the vaccine is not
protective for humans.
- Animal handlers can lower their risk by wearing protective gloves.

Lactobacillus species

• The lactobacilli are gram-positive bacilli within square ends; arranged in pairs and in
chains. They are anaerobes that can be aerotolerant and α-hemolytic; nonmotile,
catalase (-); and majority are vancomycin resistant.
• Lactobacillus species are major members of the normal microbiota of the human
vagina, gastrointestinal tract, and oropharynx.
• Glycogen deposited in vaginal epithelial cells under the influence of estrogenic
hormones is metabolized by lactobacilli to lactic acid.
• The lactic acid product of their metabolism helps maintain the low pH (pH 4 to 5) of
the normal adult female genital tract which is optimal for growth and survival of the
lactobacilli, but inhibits many other organisms.
• They rarely cause disease, and if present, they may be found occasionally in deep-
seated infections.

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MLS 223_Evaluate 5.1.

Matching type:. Match the descriptions in each item in column A with the options
provided in column B.

A B

1. Originally a disease of animals, the characteristic A. C. diphtheirae

lesions are dark, swollen, and often found on the B. C. jeikeium
hands. C. C. pseudodiphtheriticum
D. L. monocytogenes
2. Disease manifestation is associated with toxin E. E. rhusiopathiae
produced by lysogenized strains. F. Lactobacillus species

3. Infections are associated to consumption of

poultry products such as milk, cheese, ice
cream, and meat.

4. Metabolic by-products confers protection to the

host.

5. A lipid-loving bacterium that causes infection in

immunocompromised individuals.

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MLS 223_Evaluate S22

Multiple choice: Select the BEST answer.

1. For the isolation of Corynebacterium diphtheriae in culture, specimens are obtained

from the following, EXCEPT:
A. throat
B. nasopharynx
C. blood
D. skin lesions

2. Which of the following culture media contains coagulated eggs?

A. Columbia CNA
B. Tinsdale agar
C. Cystine-tellurite blood agar
D. Loeffler's slant

3. Brown halos around colonies on Tinsdale agar indicates:

A. hemolysis of red blood cells
B. tellurite reductase activity
C. cystinase activity
D. toxin production

4. Elek test is performed on a Corynebacterium diphtheriae isolate to detect:

A. diphtheria toxin expression
B. four distinct colony types
C. weakly β-hemolytic reaction of biotype mitis
D. none of the above

5. Which test/characteristic can be used to distinguish between Corynebacterium

ulcerans and Corynebacterium pseudotuberculosis?
A. Presence of polyphosphate inclusions
B. Black colonies on CTBA
C. Urease test
D. Production of acid from starch or glycogen

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References:

Carroll, K. C. (2016). Jawetz, Melnick & Adelberg's medical microbiology (27th

ed.). New York: McGraw-Hill Education.

Melnick, J. L., Jawetz, E., Adelberg, E. A., & Riedel, S. (2020). Jawetz, Melnick y
Adelberg Microbiología médica. México: McGraw-Hill.

Procop, G. W., Church, D. L., Hall, G. S., Janda, W. M., Koneman, E. W.,
Schreckenberger, P. C., & Woods, G. L. (2017). Color Atlas and Textbook of
Diagnostic Microbiology (7th ed.). Philadelphia: Wolters Kluwer Health.

Talaro, K. P., & Chess, B. (2018). Foundations in Microbiology (10th ed.). McGraw
Hill.

Tille, P. M. (2017). Bailey & Scott's Diagnostic Microbiology (14th ed.). St. Louis,
Missouri: Elsevier.

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