United States
               Environmental Protection
               Agency
National Risk Management
Research Laboratory
Cincinnati, OH 45268
               Research and Development
EPA/600/SR-95/088
September 1995
EPA      Project Summary
               Microbiological Screening of the
               Indoor Air  Quality  in the  Polk
               County Administration  Building

               D. W. VanOsdell, K. E. Leese, and K. K. Foarde
                Designing and operating a ventila-
               tion system for increased  outdoor air
               rates, as required by ASHRAE Stan-
               dard 62-1989, improves indoor air qual-
               ity  (IAQ) but  is thought to extract a
               penalty in energy costs and, potentially,
               increased microbial contamination in a
               hot humid climate. A two-part research
               program into the impact of increased
               outdoor air rates (per ASHRAE 62-1989)
               on  building microbial contamination
               and the cost of providing that outdoor
               air  was initiated by the Research Tri-
               angle Institute for the U.S. EPA.  The
               Polk County Administration Building
               (PCAB), a  large negatively pressur-
               ized  building,  not  known to  be
               biocontaminated, was selected for the
               study. The microbiological screening
               of the PCAB is the subject of this re-
               port; the energy/cost analysis is the
               subject of a separate  report.
                The  microbiological screening  in-
               cluded bioaerosol, bulk material, con-
               densate, surface,  and building  floor
               dust samples taken  at multiple  loca-
               tions. In general, the  microbial results
               were consistent with the PCAB's being
               a non-problem building. However, the
               study was too limited  in both duration
               and number of sample locations to
               completely  evaluate the building.  The
               results of a few samples indicated mi-
               crobiological conditions that might war-
               rant further investigation but were not
               of themselves  adequate to indicate a
               building-wide problem.
                This Project Summary was developed
               by  the National Risk Management Re-
               search Laboratory's Air Pollution  Pre-
               vention and Control Division, Research
 Triangle Park,  NC, to  announce key
 findings of the research project that is
 fully documented in a separate report
 of the same title (see  Project Report
 ordering information at back).

 Introduction
  This research  was an  indoor microbio-
 logical screening of the Polk County Ad-
 ministration Building (PCAB)  in Bartow,
 FL. Its goal was to generate  a baseline
 measurement that could  be used, in
 conjuction with additional sampling that
 has not been undertaken at this time, to
 evaluate the impact of ventilation system
 design and operation on the microbiologi-
 cal aspects of indoor air quality (IAQ).
  Indoor microbiological contamination can
 be a significant cause of poor  IAQ and is
 known to be associated with building ven-
 tilation systems. The impact of  a building's
 ventilation system on biocontamination is
 complicated. On the positive side,  build-
 ing  pressurization reduces the infiltration
 of biocontaminants, while maintenance of
 relatively dry indoor environmental condi-
 tions prevents the growth of the microor-
 ganism spores inside. Biocontaminants will
 grow and amplify on building materials at
 lower moisture levels than previously re-
 ported, and the appropriate level of mois-
 ture remains under investigation. Filtration
 equipment in the ventilation systems can
 similarly reduce the influx of environmen-
 tal microorganisms. On the other  hand,
 improperly designed, maintained, or oper-
 ated ventilation systems can contribute to
 indoor biocontamination.  In addition, the
 increased outdoor  air  rates called  for in
 ASHRAE Standard  62-1989 are said to
 both increase building energy requirements

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and lead to increased microbiological con-
tamination  in  hot  and humid  climates.
  The  PCAB was  not thought to have a
microbiological  (or any  other)  problem.
However, a neighboring building with some
characteristics similar to the PCAB was
known  to have had a very significant  mi-
crobial  contamination problem. The PCAB
operated  under negative pressure, which
had the potential to bring in outdoor  mi-
croorganisms. Microbiological  investiga-
tions are commonly conducted in problem
buildings, and the  mere  presence of  mi-
croorganisms in a building does  not in
itself indicate a problem. Microorganisms
are endemic in buildings, their concentra-
tions can vary widely over short periods of
time, and  the  "grab-sample" nature  of
bioaerosol samplers makes  the results of
a small number of short-term samples  dif-
ficult to analyze. The indoor and outdoor
levels and  types of organisms  must  be
interpreted  in relation to  those in nearby
buildings  or  control areas  of the  same
building. For  these reasons, a screening
study such as this one  is too limited in
both duration and number of sample loca-
tions to completely  evaluate a  building.
  During  this study, microbiological data
were collected  from bulk,  surface,  and
bioaerosol samples, and the moisture con-
tent of  some  building materials was mea-
sured.  Each  of the  measurements  ap-
proaches the  question of biocontamination
from a different perspective,  thus address-
ing the problem of  identifying the sources
of biocontamination.
  The  PCAB is a 5-story, 14,000  m2
(149,000 ft2)  brick-faced  building con-
structed in  1988. It has a permanent  oc-
cupancy of approximately 300 county em-
ployees and elected officials, and also has
a transient population who come  to  the
building to conduct  county  business.
Bartow is  in  central Florida and has  the
hot, humid climate typical of that area.
  The  heating, ventilating,  and air-condi-
tioning  (HVAC) system in the PCAB  uti-
lizes variable  air volume delivery of condi-
tioned air and a plenum return. The air is
conditioned in chilled water coils located
in variable  air volume (VAV) air handling
units (AHUs), filtered with 5-cm  (2-in.)
ASHRAE-30 filters, then  reheated as  re-
quired for delivery to the space. The air is
distributed to fan-powered VAV terminal
boxes.  The relative humidity in the  build-
ing  is  controlled to approximately  40%.
Each floor has multiple HVAC zones.

Procedure
  The screening study included a building
walk-through,   outdoor   and  indoor
bioaerosol  sampling,  bulk  and surface
sampling,  and occasional building mate-
rial moisture  measurement. All sampling
was conducted in April 1994. Bulk samples
consisted  of  HVAC  fiberglass ductliner,
condensate from  drain pans, and  com-
posite  carpet dust.  Surface  samples in-
cluded swabs from inside selected AHUs
and the back side  of ceiling tiles.  This
study  was HVAC-system driven, and the
test plan allowed some adjustment of test
sites and other aspects of the study based
on conditions in the  building.  All microbial
samples were shipped to RTI for analysis.

Test Locations
  A walk-through  of the entire building to
note any visible potential microbial  prob-
lems was  the first step of the screening
study. Study locations were chosen to be
in  both the  interior  and  exterior  HVAC
zones  and would have  been chosen to
coincide with a potential problem area had
one   been  observed.  Each  indoor
bioaerosol sample was paired with an out-
door sample  near the outdoor air intake
for the HVAC unit serving the indoor site.
The majority  of the screening  samples
were collected in  two rooms on the first
floor, two rooms on the fourth floor, and at
one location on the fifth floor.

Sampling and Inspection
  The  HVAC  systems serving  the test
zones were inspected for visible problems
(standing  water, plugged  condensate
drains, duct leaks, etc.), and samples were
obtained when appropriate.
  Cleanliness  near  the air sampling re-
gions  was evaluated qualitatively  by in-
spection and  noted  on the data sheet by
location.
  Building material moisture  content was
evaluated  when appropriate using a  con-
ductivity meter internally calibrated  and
set on the  concrete and plaster scale. The
readings are relative, and the instrument
was used  to  help  identify any moist loca-
tions that might be microbial reservoirs or
have the  potential to become microbial
sources.

Biocontaminant Sampling
  All bioaerosol sampling was conducted
in temporarily vacated offices  or after work
hours  to avoid disturbing the  PCAB occu-
pants.  Indoor  and  outdoor  air  samples
were obtained with Mattson-Garvin slit-to-
agar samplers operated for  30 minutes
indoors and  5  minutes  outdoors. Tests
were conducted at nine sites, and at each
site duplicate samples were collected se-
quentially with each of two fungal  media
and one bacterial media, for  a total of 54
Mattson-Garvin runs. At a single test site,
the total period during which sampling was
conducted was about 90 minutes indoors
and 45 minutes outdoors.
  Swab surface  and bulk material sam-
ples  were collected at appropriate loca-
tions to assess microbial flora within the
PCAB. Three microbiological media were
employed during  the analysis to  cover as
broad a range of microorganisms as pos-
sible.

Results and Discussion

Air Samples
  Table 1 summarizes the mean levels of
colony forming units (CFU)/m3 for the xe-
rophilic (low water requirement) fungi and
the bacteria  at  each of the nine sites
sampled. The results of the analyses on
the general fungal  media are not shown
but supported those for the xerophilic me-
dia. Each entry is the mean of sequential
duplicate measurements. A comparison of
the outdoor and indoor mean levels shows
that for all  the pairs  there were  less or-
ganisms isolated indoors than out. This
result is consistent with that found in a
non-problem building. However, the mean
values given in Table 1 do not show that
for Rooms  170 and 413 there were con-
siderable differences  between the results
for the two  sequential duplicates. In both
rooms, one of the two replicates was sev-
eral times higher than the other, and that
result was  observed  on both  media col-
lected  at the  same  time. Thus the  el-
evated values are not experimental error,
but true measurements of a short-term
elevated  level. This  difference  between
sequential duplicate sampling runs in the
same room suggested  that comparisons
should be made  between the runs based
on the identification of the organisms.
  Cladosporium spp. predominated in all
the outdoor samples taken at the  PCAB,
with over 50% of the total CFUs identified
as belonging  to that  genus. The second
most commonly isolated mold in the out-
door air was Penicillium with less than
25% of the total colonies. It is generally
expected that the  numbers and distribu-
tion of indoor airborne fungi  in mechani-
cally ventilated non-problem buildings will
reflect those found  in the outdoors, but at
lower levels.  In most of the indoor sam-
pling locations in the PCAB, Cladosporium
was the predominant genus followed by
Penicillium.  The duplicate samples  for
Rooms 170 and  413  were noticeably dif-
ferent in that there was a change in the
distribution  of the  predominant  fungi.  In
Room  170  there was an increase in the
Penicillium  isolated from  7  to  28%.  In
Room  413,  the  numbers were  14 and

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Table 1. Mean Total Airborne Fungi and Bacteria in CFU/m3
Location
                         Room
                                               Xerophilic Fungi
                                                                         Bacteria
1st floor NE
1st floor NE
1st floor SW
1st floor SW
4th floor NE'
4th floor NE
4th floor SW
4th floor SW
5th floor NE
5th floor NE
Outdoor Air
Room 170
Outdoor Air
Room 138A
Outdoor Air
Room 413
Outdoor Air
Room 440
Outdoor Air
Indoors
1100
610
580
110
530
210
830
80
530
30
520
330
1900
270
250
80
440
190
250
30
* The outdoor air sample collected on the 5th floor NE was paired with both the 5th floor indoor sample and the sample
 collected in Room 413 because it was near both the 4th and 5th floor outdoor air intakes.
67%.  By itself,  the 28% Penicillium spp.
isolated from Room 170 might not be ex-
cessive. However, there was also a 10-
fold increase in total counts  between the
duplicates,  suggesting that an  indoor
source  might  be present.  In  the  same
samples, the airborne concentrations of
Aspergillis spp.  also increased. For Room
413, while the counts on the second repli-
cate increased, the total level was only
332 CFU/m3 and, therefore,  not  exces-
sively high.  However, that 67% of those
were  Penicillium suggests again that  an
indoor source might be present. Although
airborne fungal measurements are grab
samples and subject to considerable vari-
ability, in both of these rooms the increase
in  Penicillium was detected by two differ-
ent samplers on two different media at the
same  time. The combined evidence indi-
cates  that potential source reservoirs of
Penicillium spp. may be contaminating the
rooms.

Surface and Bulk Samples
  A number of different surface and bulk
samples were collected: condensate from
drain  pans,  swabs of ceiling tiles and
AHUs, bulk samples of fiberglass ductliner,
and composite carpet  dust. None of these
samples showed any remarkable levels or
distribution of bacteria or fungi.
  The  other bulk  samples,  fiberglass
ductliner samples from the 4th floor AHU
and swab samples from the 1st floor south-
west AHU  and the 4th floor northeast AHU,
yielded  potentially significant numbers of
Penicillium'm practically pure culture. Swab
samples of a small  patch of white  myce-
lial-like  material were taken  in AHU2  lo-
cated  on  the  1st floor (southwest) and
AHU9 located on the 4th floor (northeast).
Analysis showed they were a pure growth
of Penicillium. Isolation of Penicillium spe-
cies from  both  the  AHU swabs and the
fiberglass  ductliner suggests that possible
source  reservoirs may have been  identi-
fied. Although speciation of the Penicil-
lium was  not performed, isolation of the
colonies in some of the AHUs is consis-
tent with the evidence  of potential  con-
tamination suggested by the air sampling,
though it does not confirm the identifica-
tion of a source reservoir.

Moisture and Cleanliness
  Moisture meter readings were taken at
a variety of locations within the building.
Only one potential water stain was identi-
fied during inspection of the building. No
readings suggested  moisture  problems.
Cleanliness was  also determined visually.
Overall, the impression of the building was
that of a clean, well-maintained facility.

Conclusions  and
Recommendations
  The overall impression of the PCAB
was of a clean,  well-maintained, low oc-
cupancy (relative to the HVAC flows) struc-
ture. The combined results of the air,  bulk,
and  surface sampling did not  indicate  a
clear biocontaminant problem in the build-
ing. On the other hand,  the sampling pe-
riod was short and samples were taken in
only a few  locations. The  elevated air-
borne levels for one  of two sequential
airborne  fungi samples in  each  of two
different rooms (confirmed by the second
fungal media), coupled with the isolation
of essentially pure Penicillium in some of
the  AHUs, give some cause for concern.
Considering that  the building is located in
a hot, humid climate, that biological con-
tamination problems have occurred in ad-
jacent county buildings (and some occu-
pants have been exposed and may  have
been sensitized to fungal contamination),
further investigation for potential  source
reservoirs might  be prudent.
  The PCAB is negatively pressurized and
appears to have  restricted outdoor air in-
takes. Infiltration  air is unfiltered and un-
conditioned, and the potential exists for
transport of biocontaminants  in the infil-
trating air, condensation of water vapor in
infiltration paths, and consequent building
contamination.  On  the  other  hand,  the
PCAB is  operated at a low relative humid-
ity that tends to prevent microbial growth,
though it is presumably expensive to op-
erate. This combination  of characteristics
presents  a number of research opportuni-
ties:
  1)The  results of this screening study
    are  not conclusive  to either identify
    the PCAB as a biocontaminated prob-
    lem  building  or  clearly  show that
    biocontamination  is not an issue in
    the  PCAB.  The study was not  de-
    signed to  accomplish that task. The
    results do show that fungi (Penicil-
    lium  spp.) may have become estab-
    lished in some AHUs and are either
    established in some parts of the ven-
    tilation system downstream of the fil-
    ters  or  at least occasionally trans-
    ported through  the  filters  to  some
    rooms at levels above those found in
    most PCAB indoor  locations and
    roughly  equivalent to outdoor levels.
    The  building may be in transition from
    non-problem to problem, and as such
    presents an  unusual opportunity to
    study some important questions, such
    as:  a)  How extensive  is the HVAC
    system contamination? b) What con-
    ditions led to that  contamination? c)
    Is  PCAB becoming a problem build-
    ing and is the  contamination  getting
    worse?  and  d) Can conditions be
    modified to prevent a serious  con-
    tamination problem from developing?
  2)The  PCAB could be modified physi-
    cally to  operate at a controlled posi-
    tive pressure to ensure that air enter-
    ing the  building is conditioned.  Both
    short- and long-term  studies of the
    microbial ecology in the building would
    provide valuable information concern-
    ing the impact of building pressuriza-
    tion  in a hot and humid climate.
  3) In combination  with pressurization,  a
    reduced-energy operating mode could
    be designed for the PCAB to, poten-
    tially, provide both reduced costs and
    reduced microbial contamination po-
    tential.
  4) In addition to building pressurization,
    the impact of building ventilation rates
    on microbial contamination  and gen-
    eral  indoor air quality could be stud-
    ied  by modifying the outdoor air in-
    takes to allow  increased  outdoor air
    delivery.  Such  operation should be
    optimized  for energy efficiency con-
    sistent with  prevention of conditions
    conducive to microbial  growth  in the
    building.

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   D. I/I/ VanOsdell, K. E. Lease, andK. K. Foarde are with Research Triangle Institute,
     Research Triangle Park, NC 27709-2194.
   Russell N. Kulp is the EPA Project Officer (see below).
   The complete report, entitled "Microbiological Screening of the Indoor Air Quality in
     the  Polk County Administration Building," (Order No.  PB95-243085;  Cost:
     $17.50, subject to change) will be available only from:
          National Technical Information Service
          5285 Port Royal Road
          Springfield, VA 22161
          Telephone: 703-487-4650
   The EPA Project Officer can be contacted at:
          Air Pollution Prevention and Control Division
          National Risk Management Research Laboratory
          U.S. Environmental Protection Agency
          Research Triangle Park, NC 27711
United States
Environmental Protection Agency
National Risk Management Research Laboratory (G-72)
Cincinnati, OH 45268

Official Business
Penalty for Private Use $300
      BULK RATE
POSTAGE & FEES PAID
         EPA
   PERMIT No. G-35
EPA/600/SR-95/088

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