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Journal of Clinical Microbiology, August 1998, p. 2377-2379, Vol. 36, No. 8
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Breast Abscess Associated with
Helcococcus kunzii
Abdul H.
Chagla,1,*
Alexander A.
Borczyk,2
Richard R.
Facklam,3 and
Marguerite
Lovgren4
London Public Health Laboratory,
London,1 and
Central Public Health
Laboratory, Toronto,2 Ontario, and
National Centre for Streptococcus, Edmonton,
Alberta,4 Canada, and
Centers for
Disease Control, Atlanta, Georgia3
Received 16 January 1998/Returned for modification 6 March
1998/Accepted 12 May 1998
 |
ABSTRACT |
Helcococcus kunzii, a nonvirulent member of the human
skin flora, has recently been implicated in causing infections in
immunosuppressed patients. We report a case of breast abscess
associated with H. kunzii in an immunocompetant patient and
discuss the criteria used in its identification and our observations of
susceptibility testing for this species.
 |
TEXT |
Helcococcus is a newly
described genus of catalase-negative, gram-positive cocci
(2). H. kunzii, the only member of this genus, is
thought to be an avirulent member of the human skin flora that is
rarely associated with wound infections (1, 5). We report
the isolation of H. kunzii from a case of breast abscess in
an immunocompetent patient and our experience with the identification and susceptibility testing of this species.
Case report.
A 57-year-old immunocompetent, nondiabetic female
presented to her physician with an infected sebaceous cyst in the skin
around the upper inner quadrant of her left breast. She was afebrile and had no systemic signs of infection. Two days prior to her presentation, she had developed local redness, swelling, and tenderness around the left breast area. The cyst was incised and drained under
local anesthesia. She was treated postsurgically with 0.5 g of
cephalexin orally every 8 h for 5 days.
A Gram stain of the cyst drainage showed numerous polymorphonuclear
leukocytes and gram-positive cocci in pairs and clumps. The pus from
the incision was inoculated onto Trypticase soy agar containing 5%
sheep blood, onto chocolate agar, onto MacConkey agar, and into cooked
meat medium. The 5% sheep blood agar and chocolate agar plates were
incubated at 36°C in a CO2-enriched (5%) atmosphere for
24 h. A separate 5% sheep blood agar plate was also inoculated
with pus and incubated anaerobically for 72 h at 36°C. Cooked
meat medium and MacConkey agar plates were incubated aerobically for
48 h at 36°C. After 24 h of incubation, a pure and heavy
growth of pinpoint, greyish, nonhemolytic colonies appeared on the 5%
sheep blood agar and chocolate agar plates incubated in
CO2-enriched (5%) atmosphere. Identical growth
characteristics were observed for the 5% sheep blood agar plate that
was incubated under anaerobic conditions. A subculture from the cooked
meat medium also yielded similar growth on 5% sheep blood agar and chocolate agar and no growth on MacConkey agar. A Gram stain of the
isolate showed gram-positive cocci in pairs and clusters. Initial spot
biochemical tests were negative for catalase and positive for
pyrrolidonyl arylamidase (PYR). A leucine aminopeptidase (LAP) test
using specific aminopeptidase substrate (diagnostic) tablets (A/S
Rosco, Taastrup, Denmark) was negative after 24 h of incubation at
36°C. Biochemical identification by use of the API 20S Strep system
(bioMerieux Vitek, Inc., Hazelwood, Mo.) resulted in profile no.
4100413 (esculin and PYR positive, LAP negative, and acid reaction from
lactose, trehalose, starch, and glycogen), indicating a doubtful
identification of Aerococcus viridans. The isolate was
identified as H. kunzii by the National Centre for
Streptococcus (NCS), Edmonton, Alberta, Canada, and confirmed by the
Centers for Disease Control and Prevention, Atlanta, Ga. The
biochemical characteristics of this strain are presented in Table
1 together with typical reactions for
eight other H. kunzii clinical isolates that have been
examined at the NCS. All strains, including the case isolate, were
further confirmed as H. kunzii by 16S rRNA sequencing done
at the AFRC Institute of Food Research, Reading, United Kingdom.
Antimicrobial susceptibility testing with various antibiotics was
performed by the disk diffusion method using Mueller-Hinton agar
supplemented with 5% sheep blood. The disk diffusion plates were
incubated at 36°C in 5% CO2 for 24 h. Based on the
recommendations made by the National Committee for Clinical Laboratory
Standards (6) for interpretive standards for
Streptococcus species, the results of the disk diffusion
assay suggested that the current isolate was sensitive to penicillin,
ampicillin, and vancomycin but resistant to erythromycin and
clindamycin. The MICs of drugs for the isolate were determined by use
of the Etest system (AB Biodisk, Piscataway, N.J.) on Mueller-Hinton
agar supplemented with 5% sheep blood. The Etest susceptibility plates
were incubated at 36°C in 5% CO2 for 24 h. The MICs
of penicillin, ampicillin, and cephalothin for the isolate were 0.12, 0.25, and 0.5 mg/liter, respectively.
Examination of the patient 1 week posttreatment revealed some residual
pus. The patient was asked to irrigate the area once
or twice daily by
allowing warm water to flow over her breast
while showering. On
reassessment after a month, she showed no
sign of tenderness, erythema,
or palpable induration. Since there
was no lump, excision of the cyst
was not done.
Weaknesses of commercial gram-positive identification systems require
the use of conventional testing media for accurate classification
of
this species. Due to close phenotypic resemblance,
Helcococcus may be misidentified as
A. viridans
(
2,
3). However unlike
Aerococcus,
Helcococcus is lipophilic (
1,
2), and this can
be
demonstrated by enhanced growth in Todd-Hewitt broth supplemented
with
0.1% Tween 80. At the NCS, we have observed that
Helcococcus strains grow best in heart infusion-based
carbohydrate media as
described by Facklam and Elliot (
3).
It is important to note,
however, that even though the
Helcococcus strains that we examined
grew well in this
medium, fermentation, as indicated by color
change, was not
demonstrated unless this medium was supplemented
with 0.1% Tween 80. We do not use bovine serum as a medium supplement
for this genus since
it may cause a color change in the base and
make interpretation
difficult. The CDC has not observed fermentation
in the carbohydrate
media for the case isolate or for other
Helcococcus isolates
that they have tested since they do not supplement the
media with 0.1%
Tween 80 or bovine serum. The fermentation of
inulin and the failure to
ferment trehalose in conventional media
observed for the case isolate
are atypical for this species both
in our experience and in previous
reports (
2,
8). Laboratories
using the API 20S Strep system
(
2,
4,
8) have reported
this species as inulin negative and
an acidifier of trehalose,
but one investigator (
4) also
noted acidification of inulin
for an isolate which they called
"
Helcococcus-like." Failure to
ferment trehalose in
unsupplemented heart infusion-based media
has been reported previously
(
2). Acidification of inulin and
failure to ferment
trehalose in Tween 80-supplemented heart infusion-based
media for our
case isolate were reproducible in repeat testing,
and these atypical
reactions may reflect our limited knowledge
of this newly described
organism.
H. kunzii should be considered a possible identification for
any isolate that grows on blood agar as tiny, nonhemolytic or
very
slightly alpha-hemolytic, grey, translucent colonies which
are catalase
negative and vancomycin susceptible and which stain
as gram-positive
cocci arranged in pairs or clusters. Demonstration
of enhanced growth
in broth supplemented with 0.1% Tween 80 offers
further support for
this identification. Like other reports (
2,
4,
8),
biochemical testing of our isolate resulted in API
profile no. 4100413. Even though
H. kunzii is not currently in
the API database,
laboratories using this system may find this
profile number useful for
preliminary identification.
Susceptibility testing of
Helcococcus is complicated by its
lipophilic nature and growth requirement for hemin, as demonstrated
by
the inability to synthesize porphyrin (
2). At the NCS, we
have observed that this genus grows very poorly, or not at all,
in
Mueller-Hinton broth, even when the broth is supplemented with
3%
lysed blood. The difficulties encountered when broth-based
susceptibility testing systems are used have been noted by others
(
1). Susceptibility testing by Etest has been used by some
laboratories (
1,
8). At the NCS we have applied National
Committee for Clinical Laboratory Standards testing criteria for
Streptococcus to
Helcococcus species
(
7). Our experience suggests
that agar dilution on
Mueller-Hinton agar supplemented with 5%
sheep blood provides the most
reliable method for MIC testing
of drugs for this species. By this
method, the MICs of penicillin
and ampicillin for our case isolate were
both 0.25 mg/liter, which
is similar to those produced by the Etest.
However, the MIC of
cephalothin was 2.0 mg/liter, fourfold higher than
the Etest MIC
that was originally reported. Resistance to erythromycin
(MIC
> 2.0 mg/liter) was confirmed by agar dilution for the case
isolate,
but the result for clindamycin was susceptible (MIC = 0.12 mg/liter).
It should be noted that of nine
Helcococcus
isolates currently
in the NCS collection, eight are resistant to
erythromycin (MIC

1.0 mg/liter) and all are susceptible to
clindamycin at MICs
of

0.25 mg/liter.
Aerococcus and
Helcococcus are considered to be
part of the normal flora of the human skin; however, unlike
Aerococcus, which
is present in the environment, the natural
habitat of
Helcococcus is not well characterized
(
9). In humans, these organisms can
cause opportunistic
infections such as endocarditis, bacteremia,
meningitis, and wound
infections (
1).
H. kunzii can cause wound
infections in immunosuppressed patients; however, in the present
case,
the patient was immunocompetent. Peel and coworkers (
8)
have
reported the isolation of
H. kunzii in pure culture and
heavy
growth from an infected sebaceous cyst on the shoulder of a male
patient. These investigators have suggested that
H. kunzii
can
act as an opportunistic pathogen. Recently, Haas and coworkers,
in
a study of skin colonization by
H. kunzii in diabetic and
nondiabetic
subjects, have suggested that this species, in addition to
being
a colonizer of human skin, may also be a previously unrecognized
member of the polymicrobic flora of lower-extremity ulcers
(
4).
Accurate identification of
H. kunzii is
important when investigating
the role
H. kunzii as a skin
colonizer and a potential pathogen.
Furthermore, our observations
suggest that resistance to erythromycin
may be expected for this
species. Our finding provides further
support for the opportunistic
role of
H. kunzii in causing infection
in both
immunosuppressed and immunocompetent patients.
 |
ACKNOWLEDGMENTS |
We thank M. D. Collins at ARFC UK for the 16S rRNA sequence
testing and P. Rawte for his technical assistance.
 |
FOOTNOTES |
*
Corresponding author. Mailing address: London Public
Health Laboratory, P.O. Box 5704, Terminal A, London, Ontario, Canada N6A 4L6. Phone: (519) 455-9310. Fax: (519) 455-3363. E-mail:
chaglaa{at}gov.on.ca.
 |
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|
Journal of Clinical Microbiology, August 1998, p. 2377-2379, Vol. 36, No. 8
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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