MODULE Enterococcus
Microbiology
17
Notes
ENTEROCOCCUS
17.1 INTRODUCTION
Enterococcus is a genus of lactic acid bacteria. Enterococci are catalase negative
Gram-positive cocci that often occur in pairs (diplococci) or short chains, and are
difficult to distinguish from streptococci on physical characteristics alone.
OBJECTIVES
After reading this lesson, you will be able to:
z describe Enterococci
z identify Enterococci in the laboratory
z explain the pathogenesis
z discuss the antibiotic resistance
17.2 ENTEROCOCCI
Enterococci are facultative anaerobic organisms, i.e., they are capable of cellular
respiration in both oxygen-rich and oxygen-poor environments. Though they are
not capable of forming spores, enterococci are tolerant of a wide range of
environmental conditions: extreme temperature (10-45°C), pH (4.5-10.0) and
high sodium chloride concentrations. They are part of the normal enteric flora.
Two species are common agents causing infection among humans: E. faecalis
(90-95%) and E. faecium (5-10%). Rare clusters of infections occur with other
species, including E. casseliflavus, E. gallinarum, and E. raffinosus. The
enterococci were previously classified as group D streptococci.
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Enterococcus MODULE
Microbiology
Notes
Fig. 17.1
Enterococci can exhibit beta/gamma-haemolysis on sheep’s blood agar.
Identification: Because the group D cell wall specific antigen is a teichoic acid,
it is not an antigenically good marker; enterococci are usually identified by
characteristics other than immunologic reaction with group-specific antisera.
z They are usually non-haemolytic, but occasionally á-haemolytic.
z The enterococci are gram positive cocci, occuring in singles, pairs, and short
chains.
z Cells are sometimes coccobacillary when gram stains are prepared from
agar plate growth. Cells are more oval and in chains when gram stains are
prepared from thioglycolate broth.
z The enterococci are facultative anaerobe and optimum growth occurs at
35°C.
z They grow well at between 10°C and 45°C whereas streptococci generally
grow at a much narrower temperature range.
z All strains grow in broth containing 6.5% NaCl and hydrolyze esculin in the
presence of 40% bile salts (bile-esculin medium).
z Enterococci hydrolyze pyrrolidonyl-ß-naphthylamide (PYR), the exceptions
to this are E. cecorum, E. columbae, and E. saccharolyticus.
z Most strains produce leucine aminopeptidase (LAP). Some strains belonging
to Group I enterococci give negative LAP tests.
z They are more resistant to penicillin G than the streptococci, and rare
isolates have plasmids that encode for â-lactamase.
z Many isolates may be vancomycin-resistant.
MICROBIOLOGY 185
MODULE Enterococcus
Microbiology
z Presumptive identification of a catalase negative gram positive cocci as an
Enterococcus can be accomplished by demonstrating that the unknown
strain is PYR and LAP positive, and grows in 6.5% NaCl and at 45°C.
INTEXT QUESTION 17.1
Notes
1. Enterococci are .............. cocci
2. Culturally Enterococci are .............. anerobes
3. Enterococci most frequently cause .............. Infection
4. .............. is highly antibiotic resistance
17.3 SPECIES IDENTIFICATION
Once established that the unknown catalase-negative gram-positive coccus is an
Enterococcus the tests can be used to identify the species. The species are
divided into 5 Groups based on the reactions in acid formation in mannitol,
sorbitol, and sorbose broths and hydrolysis of arginine.
The pigmentation test aids in the identification of E. casseliflavus, E. mundtii, E.
pullins, E. gilvus and E. sulfureus. These enterococci produce a yellow pigment
that can be detected on several different media.
The pyruvate utilization test aids in the differentiation of E. faecalis and E.
faecium. This test is also used to help differentiate between E. faecalis variant
strains and E. hirae.
The tellurite tolerance test aids in the differentiation of E. faecalis and E.
faecium.
E. haemoperoxidus is variable in the mannitol reaction and may be in group II
or III.
Diseases caused: There are at least 12 species of enterococci. Enterococcus
faecalis is the most common and causes 85–90% of enterococcal infections,
while Enterococcus faecium causes 5–10%, while E. faecalis remains the
predominant species in clinical infection, [Link] isolates are increasing in
proportion. The trend is particularly true for blood isolates.
The enterococci are among the most frequent causes of nosocomial infections,
particularly in intensive care units, and are selected by therapy with cephalosporins
and other antibiotics to which they are resistant.
186 MICROBIOLOGY
Enterococcus MODULE
Enterococci are transmitted from one patient to another primarily on the hands of Microbiology
hospital personnel, some of whom may carry the enterococci in their
gastrointestinal tracts. Enterococci occasionally are transmitted on medical
devices.
In patients, the most common sites of infection are the urinary tract, wounds,
biliary tract, and blood. Enterococci may cause meningitis and bacteraemia in
neonates. In adults, enterococci can cause endocarditis. Notes
However, in intra-abdominal, wound, urine, and other infections, enterococci
usually are cultured along with other species of bacteria, and it is difficult to
define the pathogenic role of the enterococci.
Antibiotic Resistance
A major problem with the enterococci is that they can be very resistant to
antibiotics. E faecium is usually much more antibiotic-resistant than E faecalis.
Intrinsic Resistance
Enterococci are intrinsically resistant to cephalosporins, penicillinase-resistant
penicillins, and monobactams. They have intrinsic low-level resistance to many
aminoglycosides, are of intermediate susceptibility or resistant to
fluoroquinolones, and are less susceptible than streptococci (10- to 1000-fold) to
penicillin and ampicillin. Enterococci are inhibited by â-lactams (eg, ampicillin)
but generally are not killed by them.
Vancomycin Resistance
The glycopeptide vancomycin is the primary alternative drug to penicillin (plus
an aminoglycoside) for treating enterococcal infections. Enterococci that are
resistant to vancomycin have increased in frequency. Vancomycin resistance has
been most common in E faecium, but vancomycin-resistant strains of E faecalis
also occur.
INTEXT QUESTIONS 17.2
1. Bile aesculin positive Gram positive coccus is ...............
2. Vancomycin resistant catalase negative gram positive cocci can be ...............
3. Two important species of Enterococci are ............... & ...............
4. Most common route of transmission of Enterococci from one to another is
by ...............
MICROBIOLOGY 187
MODULE Enterococcus
Microbiology
WHAT YOU HAVE LEARNT
z Enterococci are catalase negative Gram-positive cocci
z Enterococci can exhibit beta/gamma-haemolysis on sheep’s blood agar
z They grow well at between 10°C and 45°C
Notes z All strains grow in presence of 6.5% NaCl and are bile-aesculin test positive
z Most strains are PYR and LAP positive
z They can be very much resistant to antibiotics
TERMINAL QUESTIONS
1. Define genus enterococci?
2. What are the diseases caused by enterococci?
3. How will you identify Enterococcus in laboratory?
ANSWERS TO INTEXT QUESTIONS
17.1
1. Gram positive
2. Facultative anaerobes
3. Nosocomial
4. E. Faecium
17.2
1. Enterococcus
2. Enterococcus
3. E. faecalis and E. Faecium
4. Contaminated hands
188 MICROBIOLOGY