MICROORGANISMS

K C C A P U L O N G | Z H E R R E R A | K D I N F A N T E
I C P A C I A | J P S E V I L L A
M C B 1 5 0 M I C R O B I A L E C O L O G Y
IN
OBJECTIVES:
Identify different microorganisms present in the air;
Understand and demonstrate the sedimentation
procedures, the gravity plate method and impingement
technique;
Compare the number and types of airborne
microorganisms present in the intramural and
extramural environments;
Learn to compute for the determination of the microbial
load present in the air using the formulas given.

GRAVITY PLATE METHOD
also called as settle plate
method
plates were exposed in the
assigned areas for 15 minutes
microorganisms were collected
by sedimentation or by gravity
to deposit air particles
(microorganisms) on the plates
ADVANTAGES
inexpensive and easily performed
very useful for directly monitoring airborne contamination of
specific surfaces
Useful for qualitative analysis of airborne microorganisms
very limited since this method is only capable of
monitoring viable biological particles that sediment into
air and settle at time of exposure
it will not detect smaller particles or droplets suspended in
the air and it cannot sample specific volumes of air
vulnerable to interference and contamination from non-
airborne sources and growth medium might deteriorate
if exposed for too long
microorganisms may easily become overgrown in heavily
contaminated conditions
DISADVANTAGES
The laboratory is the area expected to have the most number of
microorganisms since microorganisms in air are most likely to
spread in areas where pathogens are handled for research
purpose (Pepper et al., 2014).


IMPINGEMENT METHOD
When air is dispersed in the liquid, particles in the air are trapped.
The usual volume of collection medium is 20 mL and the typical
sampling duration is approximately 20 minutes which prevents e
vaporation during the sampling of warm climates or freezing of
the liquid medium when sampling at lower temperatures.
Quantification of airborne microorganisms is accomplished by
plating the collection fluid.
A suitable collecting medium for liquid impingement samplers
must preserve the viability of the microorganism while
inhibiting its multiplication.

ADVANTAGES
gives quantitative results where sample volume can be
calculated using the flow rate and sampling time
more accurate representation of present microorganisms in air
will be obtained

DISADVANTAGES
the technique may damage some microbial cells and affect
viability
no particle size discrimination which prevents accurate
characterization of the sizes of the airborne particles that are
collected, and at overlong sampling times it allow
multiplication of cells in the liquid medium
DISCUSSIONS
More bacterial air spores grew on PCA compared to that of
the NA plate mainly because PCA has glucose, making
it richer in terms of nutrient content and carbon source.
(In samples obtained from lab and rooftop).
The glass beads acted as dispersants. They
were used to prevent bubble
formation which could have affected the microbial cell dif
fusion. It breaks aggregates of microorganisms thus
uniformly releasing them into the solution and further
trap the microorganisms in the solution because of its
pores.

RESULTS
SITE IMPINGEMENT TECHNIQUE GRAVITY PLATE METHOD
bacteria (PCA) fungi (PDA) NA bacteria
(PCA)
fungi
(PDA)
10
0
10
-1
10
0
10
-1

Lab 21,68 52,81 0,1 1,1 58,55 68,29 0,4
Street 86,121 122,60 1,3 2,0 50,49 32,28 5,13
Rooftop 86,144 65,spreader 2,0 1,0 17,15 26,34 1,3
Table 3.1. Bacterial and fungal counts recorded from air sampled in
different environments.
Table 3.2. Computed microbial load (cfu / ft
3
air) for the various
environments sampled.






* sampling time= 20 mins; ft
3
of air sampled = 7.1429; V
i
= 50mL


SITE cfu / mL of impingement liquid cfu / ft
3
air*
bacteria fungi bacteria fungi
Lab 1.68 x 10^3 <100 ESPC 1.18 X 10^4 <700
Street 1.77 x 10^3 <100 ESPC 1.24 X 10^4 <700
Rooftop 1.40 x 10^3 <100 ESPC 9.80 x 10^3 <700
RESULTS
Table 3.3. Cultural and morphological characteristics of predominant
bacteria and fungi present in air sampled from different environments.
SITE DOMINANT BACTERIA DOMINANT FUNGI
cultural
(PCA)
microscopic
(OIO)
cultural
(PDA)
microscopic
(HPO)
Lab Circular, white, flat,
entire margination
G(-), cocci cottony, circular,
white
filamentous
Street White colony
(spreader), opaque
G(-), long rods, in
chains

Green mold, cottony,
compact
conidia borne on a
vesicle, septate,
possibly Aspergillus
Rooftop circular, entire margin,
flat, yellow-pigmented
colony
G(+), circular shape a) cottony,
filamentous form and
margin, white
club-shaped
conidiophore, septate,
possibly Aspergillus
RESULTS

COMPUTATIONS
cfu / ft
3
air

Given:
sampling time = 20min
capacity of limiting orifice = 10 L / min
volume of impingement liquid after
sampling = 50mL




Number of liters per sample = [sampling time (in
minutes)] x [capacity of limiting orifice]
= 20min x 10 L / min
= 200 L

Cubic feet of air sample = liters of sample / 28
= 200 L / 28
= 7. 1429 ft
3



FOR BACTERIA
CFU per cubic feet of air =
N Vi
Va

N= CFU/ml of impingement
V
i
= 50ml
V
a
= 7. 1429 ft
3
1.68 10
3
(50)
7.1429
3
= 1.18 10
4

1.77 10
3
(50)
7.1429
3
= 1.24 10
4

1.40 10
3
(50)
7.1429
3
= 9.80 10
3

Laboratory

Street

Rooftop

FOR FUNGI

=
< 100 (50)
7.1429 ft
3

= < 700
= < 100
REFERENCES
Compendium of Methods for the Microbiological Examination of Foods (Frances
Pouch Downes, Keith Ito) American Public Health Association, Apr 1, 2001

Food Quality Magazine: Air Sampling 101. Judie Buddemeyer. June/July 2005 issue.
http://www.foodquality.com/details/article/878155/Air_Sampling_101.html?tzcheck=1

Pepper, I.L., Gerba, C.P. and Gentry, T.J. 2014. Environmental Microbiology. 3
rd
edition.
Londom: Elsevier Inc.