CHAPTER 3

Air Sampling Techniques

1. Introduction

To investigate biological particles in the air, they have to be caught by a sampling device
with a design format suitable for viewing particles under a microscope or other form of
analysis. Although quantitative analytical methods such as immunological and
molecular techniques (real-time PCR), which can quantify target DNA accurately, are
now available, this book is primarily concerned with the visual identification of airbor-
ne biological particles. As we have discussed in previous chapters, airborne particles vary
greatly in size, number and type depending on time of day, weather, season and geogra-
phical location. As a result, the location of a sampling site, time and duration of opera-
tion has to be considered carefully to ensure a representative sample is taken.
Furthermore, the type of sampling device used is important as each has different samp-
ling efficiencies or suits a particular form of analysis.
Particles can either settle onto a surface by gravitational sedimentation or be impac-
ted. Air sampling techniques described in this chapter and relying on these principles
include sticky microscope slides (or other surfaces), the cascade impactor, the Andersen
sampler, the whirling arm trap, the Burkard trap and the cyclone.
Much work on the sampling of the air spora has been done during the last 50 years,
with methods and samplers described in the Bioaerosols Handbook (Cox and Wathers,
1995). Hirst (1995) in his introduction describes a bioaerosol ‘as an aerosol comprising
particles of biological origin or activity which may affect living things through infectivi-
ty, allergenicity, toxicity, pharmacological or other processes. Particle sizes may range
from aerodynamic diameters of ca. 0.5 to 100 μm.’ In his chapters on ‘Inertial
Samplers’, and ‘Non-inertial Samplers’ Crook (1995a and b) describes many samplers
that are mainly used indoors and often testing for viable microorganisms. Outdoor air
sampling techniques are described by Lacey and Venette (1995). The simplest samplers
collects spores passively, by sedimentation onto sticky slides or agar media, or by impac-
tion due to air currents onto thin vertical sticky surfaces. A common feature of active air
samplers is the principle of impaction. Impactors use inertial impaction to capture par-
ticles on surfaces. The surfaces can be coated in a “sticky” film to retain the particles as
with the cascade impactor, whirling arm trap and Burkard spore trap, or it can be a cul-
ture medium as in an Andersen sampler. Impactors are usually fairly low volume sam-
plers e.g. the Burkard spore trap which samples at 10 l min-1, but some e.g. the VWR
(formerly Merk) MAS 100 Air Sampler and the Oxoid MAQS II microbiological air

AIR SAMPLING TECHNIQUES 35

 sampler, have sample rates as high as 100 and 120 l min-1 respectively. The size collecti-
on efficiency of inertial impactors depends on the speed of the airflow through the inlet,
the width of the inlet and the separation between the inlet and the collecting surface.
The Burkard spore trap for instance has a sample cut-off at about 1-2 μm as particles
smaller than this are able to change direction and remain within the airflow. For long-
term sampling there may be problems with sample loss through re-suspension or boun-
ce-off, unless the sticky surfaces are moved past the air intake or orifice (as with the
Hirst or Burkard spore traps), or the collection surfaces are continuously washed with
liquid (as with wet-cyclone samplers and the SKC BioSampler. For collection onto solid
surfaces the choice of collection medium is important as its efficacy depends on the
ambient temperature. If the collecting surface is a clear film or glass slide coated with a
sticky substance then the sample can be examined by microscopy.
Impaction samplers are available using a narrow slit to direct the airstream onto a
clear glass slide coated with a sticky substance in a preloaded sterile cassette, e.g. the
VersaTrapTM (produced by SKC, Dorset), can be used at flow rates of 5-30 l min-1 and
is analysed by microscopy. Particles can impact on falling raindrops and therefore rain-
water can be sampled for particles impacted on the raindrops as they fell or originating
as the nucleus around which the raindrop formed.
In some recently developed formats, the sample collection surface or collecting
medium is compatible with DNA analysis or immunological techniques e.g. miniature
cyclone sampler (Emberlin and Baboonian, 1995), and the Burkard microtiter immu-
nospore trapping device (MTIST) (Kennedy et al., 2000)]. Even established formats
such as the Burkard spore sampler can allow samples to be extracted and analysed using
molecular methods (Calderon et al., 2002).

2. Passive traps

This is the simplest way of collecting airborne biological particles. To trap particles by
sedimentation, a microscope slide is made sticky by coating one side of it with petro-
leum jelly, glycerine jelly or silicone grease. The slide is placed horizontally with the stic-
ky surface upwards. As an alternative, Petri dishes containing selective medium may be
left open to sample viable propagules. These are usually exposed for 10-30 minutes but
possibly longer or much shorter durations would be appropriate depending on the envi-
ronment. For outdoor use, a rain-shield may be mounted above the coated-slide or Petri
dish.
The technique is less efficient for small particles as due to Stoke’s Law, large particles
settle more quickly and consequently are deposited preferentially. The rate of settling, S,
is proportional to the particles’ concentration in the air (S=Cvd) where the constant of
proportionality, vd , is the velocity of deposition (Chamberlain, 1975). In non-turbulent
air, vd is the same as the terminal velocity, vt. however in turbulent air, e.g. within a crop
canopy, vd is often greater than vt (McCartney and Fitt, 1985) and vd is affected by par-
ticle size with the ratio of vd / vt for spores trapped on horizontal slides over a barley
crop decreasing from about 6 for spores with vt = 0.1 cm s-1, to about 3 for spores with

36 THE AIR SPORA