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Journal of Clinical Microbiology, July 2005, p. 3244-3246, Vol. 43, No. 7
0095-1137/05/$08.00+0 doi:10.1128/JCM.43.7.3244-3246.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.
Comparative Recovery of Microorganisms from BacT/ALERT Plastic and Glass FA and FN Blood Culture Bottles
J. A. Riley,
B. J. Heiter, and
P. P. Bourbeau*
Geisinger Medical Center, Danville, Pennsylvania
Received 23 December 2004/
Returned for modification 25 January 2005/
Accepted 9 March 2005

ABSTRACT
bioMerieux, Inc., has recently introduced plastic bottles to
replace glass bottles for use in the BacT/ALERT blood culture
system. We compared the performance of the plastic to the glass
bottles in a large clinical evaluation. Two blood cultures were
collected from each patient, one using glass FA (aerobic) and
FN (anaerobic) bottles and one using plastic FA and FN bottles.
Of the 4,040 sets of four bottles collected, 3,110 contained
the recommended 8 to 12 ml of blood, yielding 524 microorganisms
with 359 judged to be clinically significant. Of the 359 significant
organisms, 255 were recovered in either one or two bottles from
both pairs of bottles in a set while 56 organisms were recovered
only from the glass bottles and 48 were recovered only from
the plastic bottles (
P, not significant [NS]). Of the 286 significant
organisms recovered only in the FA bottles (glass and plastic),
180 were recovered in both bottles, 57 in the plastic bottles
only, and 49 in the glass bottles only (
P, NS). Of the 303 significant
organisms recovered in the FN bottles only (glass and plastic),
212 were recovered in both bottles, 46 in the plastic bottles
only, and 45 in the glass bottles only (
P, NS). For individual
organisms, the only significant difference in recovery was obtained
for
Escherichia coli, with more isolates recovered in the FN
plastic than in the FN glass bottles (
P = 0.02). These data
suggest that recovery of microorganisms with plastic FA/FN bottles
is at least equal to that with glass FA/FN bottles while offering
greater safety for users.

INTRODUCTION
bioMerieux, Inc. (Durham, NC), recently introduced plastic blood
culture bottles to replace the previously utilized glass blood
culture bottles. This change was made to reduce or eliminate
the possibility of breakage. The purpose of this study was to
compare the performance of the glass and plastic aerobic (FA)
and anaerobic (FN) bottles with a full 10-ml blood draw in each
bottle.

MATERIALS AND METHODS
The standard of care for adult patients at Geisinger Medical
Center is two pairs of blood cultures collected sequentially
via separate venipunctures or separate line access. For this
study, sets of four BacT/ALERT blood culture bottles were specially
prepared in the laboratory. Sets included one each of a plastic
FA (PFA), plastic FN (PFN), glass FA (GFA), and glass FN (GFN)
blood culture bottle. Each pair of PFA-PFN or GFA-GFN bottles
was labeled with the number one or two in order to facilitate
collecting first into the PFA-PFN pair for approximately one-half
of the patients while the remainder of the patient draws were
collected first into the GFA-GFN pair. The standard Geisinger
Medical Center protocol was followed for skin preparation prior
to collection. Blood volumes were verified (8 to 12 ml) by the
use of a visual standard.
For the sake of clarity, the following terminology is consistently utilized in this report: the term "set" refers to all four bottles collected during a patient episode, the term "pair" refers to two bottles collected by a single collection procedure, and the term "bottle" refers to a single bottle. Hence, there are two bottles in a pair and two pairs in a set. All bottles were incubated in BacT/ALERT blood culture cabinets at 35°C, using a 5-day protocol. Culture workup and identification were performed using the standard Geisinger Medical Center microbiology laboratory procedures. All cultures were worked up and reported by the standard protocols used by the microbiology laboratory at Geisinger Medical Center. A 5-day incubation period was used for all negative bottles.
If one or more bottles from a set gave a positive signal by the instrument and there was growth upon subculturing, any remaining bottles from the set that were instrument negative after 5 days of incubation were subcultured.
For the purposes of this study, isolates of coagulase-negative staphylococci, Bacillus spp., viridans group streptococci, Propionibacterium spp., and aerobic diphtheroids isolated from only one pair of bottles in a set were considered not to be clinically significant. Statistical analysis was performed as described by Ilstrup (1).

RESULTS
A total of 4,040 sets of four bottles were collected. Of these
4,040 sets, 3,110 contained the recommended 8 to 12 ml of blood
in each of the four bottles. These 3,110 sets yielded 524 microorganisms,
including 359 judged to be clinically significant and 165 judged
to be probable contaminants. Of the 359 significant organisms,
255 were recovered in either one or two bottles from both pairs
of bottles in a set while 56 were recovered only from the glass
bottles and 48 were recovered only from the plastic bottles
(
P > 0.05; not significant [NS]) (Table
1). Separate analyses
were performed to compare the performances of the two aerobic
(PFA and GFA) and the two anaerobic (PFN and GFN) bottles. Of
the 286 significant organisms recovered in the FA bottles (both
glass and plastic), 180 were recovered in both bottles, 57 in
the PFA bottles only, and 49 in the GFA bottles only (
P, NS)
(Table
2). Of the 303 significant organisms recovered in the
FN bottles (both glass and plastic), 212 were recovered in both
bottles, 46 in the plastic bottles only, and 45 in the glass
bottles only (
P, NS) (Table
3). For individual organisms, the
only significant difference in recovery was obtained for
Escherichia coli, with more isolates recovered in the plastic FN bottles
(
P = 0.02). There were no significant differences in the number
of probable contaminants isolated between the glass and the
plastic bottle sets overall or between the aerobic bottles.
However, more contaminants were recovered from the plastic anaerobic
bottles than were recovered from the glass anaerobic bottles
(
P = 0.03).
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TABLE 2. Comparative yield of 388 isolates from plastic aerobic (FA) and glass aerobic (FA) blood culture bottles by bottle type
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TABLE 3. Comparative yield of 391 isolates from plastic anaerobic (FN) and glass anaerobic (FN) blood culture bottles by bottle type
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Subculture of bottles that were reported as negative by the
instrument but had a companion positive bottle from the set
detected by the instrument yielded 17 organisms. There were
10
Pseudomonas aeruginosa isolates (5 from PFN and 5 from GFN
bottles) and 1
Alcaligenes sp. isolate (PFN) detected by subculture
from an anaerobic bottle of a pair that had a positive aerobic
bottle. One isolate of
Eubacterium lentum was detected in a
PFN bottle but not in the GFN bottle, while one isolate of
Pseudomonas aeruginosa was instrument positive in a PFA bottle but detected
only by subculture from the GFA bottle. One
Alcaligenes sp.
isolate was detected by subculture of the PFN bottle, while
the GFA bottle from the set was instrument positive for an isolate
of
Klebsiella pneumoniae and the GFN bottle from the same set
was instrument positive for an isolate of
Serratia marcescens.
Two
Propionibacterium sp. isolates and one
Corynebacterium sp.
isolate were detected by subculture of bottles while each of
the paired bottles was instrument positive for isolates of a
Staphylococcus sp. other than
Staphylococcus aureus.

DISCUSSION
Since the introduction of the original BacT/ALERT blood culture
system, several "generations" of new bottles have been introduced.
Initially, only bottles with the designations "standard aerobic"
and "standard anaerobic" were available; at a later time, additional
bottles with the designations "FAN aerobic" and "FAN anaerobic"
were introduced. The FAN bottles were designed to enhance the
recovery of fastidious bacteria, bacteria from patients receiving
antimicrobial therapy, and yeasts in comparison to recovery
with the standard BacT/ALERT blood culture bottles (
4,
5). Subsequently,
nonvented BacT/ALERT bottles, designated FA and FN, replaced
the FAN aerobic and FAN anaerobic blood culture bottles. The
FA and FN bottles have a different medium formulation and a
different type of sensor than the FAN bottles (
2). The FA bottles
contain a different concentration of charcoal, a smaller volume
of medium, and more headspace gas than the FAN aerobic bottles.
Lastly, the FA bottles are unvented, while the FAN aerobic bottles
required transient venting.
In addition to the obvious change from glass to plastic in the bottles, the plastic bottle necessitated a change in the liquid sensor used in the bottles (Steven Rothenberg, personal communication). No changes were made in media or atmosphere in the bottles. Consequently, it is not unexpected that there would be little if any difference in recovery between the glass and plastic bottles. Petti et al. reported no difference in recovery between plastic and glass PF bottles (3).
The plastic bottles weigh approximately 4 pounds less per 100 bottles than glass bottles. In our laboratory, which performs around 25,000 blood cultures per year, use of plastic bottles results in approximately 2,000 pounds less of biohazard waste per year than use of glass bottles. The potential cost savings for biohazard waste disposal would be determined by disposal costs for each laboratory.
When we routinely used glass bottles in our laboratory, we periodically received a bottle that was cracked during transit to the laboratory. Because the glass bottles had a plastic wrapper to curtail shattering if dropped, cracks that developed as a result of dropping were not always visually apparent. On several occasions, we did not detect cracked bottles until after they were loaded into the instruments. Subsequent leaks required decontamination of individual cells and, on one occasion, decontamination of an entire block. This potential problem should be eliminated with plastic bottles.
In conclusion, in our evaluation, which included 3,110 blood culture sets, we found no significant difference in recovery of clinically significant microorganisms between the glass and plastic FA and FN bottles, although the plastic bottles did recover more organisms. Moreover, plastic bottles offer greater safety and, potentially, lower disposal costs than glass bottles.

FOOTNOTES
* Corresponding author. Mailing address: Division of Laboratory Medicine, Geisinger Medical Center, Danville, PA 17822-0131. Phone: (570) 271-7467. Fax: (570) 271-6105. E-mail:
pbourbeau{at}geisinger.edu.


REFERENCES
1 - Ilstrup, D. M. 1978. Statistical methods employed in the study of blood culture media, p. 31-39. In J. A. Washington (ed.), The detection of septicemia. CRC Press, West Palm Beach, Fla.
2 - Mirrett, S., R. J. Everts, and L. B. Reller. 2001. Controlled comparison of original vented aerobic FAN medium with new nonvented BacT/ALERT FA medium for culturing blood. J. Clin. Microbiol. 39:2098-2101.[Abstract/Free Full Text]
3 - Petti, C. A., S. Mirrett, C. W. Woods, and L. B. Reller. 2005. Controlled clinical comparison of plastic versus glass bottles of BacT/ALERT PF medium for culturing blood from children. J. Clin. Microbiol. 43:445-447.[Abstract/Free Full Text]
4 - Weinstein, M., S. Mirrett, L. Reimer, M. Wilson, S. Smith-Elekes, C. Chuard, K. Joho, and L. Reller. 1995. Controlled evaluation of BacT/Alert standard aerobic and FAN aerobic blood culture bottles for detection of bacteremia and fungemia. J. Clin. Microbiol. 33:978-981.[Abstract]
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Journal of Clinical Microbiology, July 2005, p. 3244-3246, Vol. 43, No. 7
0095-1137/05/$08.00+0 doi:10.1128/JCM.43.7.3244-3246.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.
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