Chapter 5 - Aeromicrobiology

Objectives

1. Be able to discuss and give examples of microorganisms of concern

in disease, biological warfare and closed environments that may be
spread through aerosols
2. Be able to describe the three components of the aeromicrobiological
pathway: launching, transport, and deposition
3. List factors that impact microbial survival in bioaerosols
4. Describe how microorganisms in air are measured
5. Describe how bioaerosols can be controlled
6. Describe the biosafety levels used in research and medicine.
Up to 70% of all plant disease is airborne.

Fungal plant disease Pathogen

Dutch Elm disease Ceratocystis ulmi
Potato late blight Phytophthora infestans
Leaf rust Puccinia recondite
Loose smut of wheat Ustilago tritici
Downy mildew Pseudoperonospora humuli
Maize rust Puccinia sorghi
Powdery mildew of barley Erysiphe graminis
Southern corn leaf blight Helminthosporium maydis
Example: Late blight of potato caused 1845 epidemic that spread from Belgium
(mid-June) throughout Europe by mid-October.
Famine related deaths 750,000 to 1,000,000.
Population of Ireland went from 8,200,000 to 4,400,000 from 1840 to 1911.

Norway
Sweden

Ireland
UK
Poland
Belgium

France Germany

Italy
Spain
 Important Airborne Pathogens

• Animal Disease
– Bacterial diseases
• Tuberculosis (Mycobacteriumm bovis)
• Brucellosis (Brucella spp.)
– Fungal diseases
• Aspergillosis (Aspergillus spp.)
• Coccidioidomycosis (Coccidiodes immitis)
– Viral diseases
• Influenza (Influenza virus)
• Rabies (Rhabdoviridae)
• Foot-and-mouth disease (Aphthovirus)
 Important Airborne Pathogens
• Human disease
– Fungal diseases
• Aspergillosis (Aspergillus fumigatus)
• Coccidioidomycosis (Coccidiodes immitis)
– Bacteria diseases
• Pulmonary tuberculosis (Mycobacterium tuberculosis)
• Pneumonia (Klebsiella pneumoniae)
• Pulmonary anthrax (Bacillus anthracis)
• Legionellosis (Legionella spp.)
• Whooping cough (Bordetella pertussis)
• Diphtheria (Corynebacterium diphtheriae)
– Viral diseases
• Influenza (Influenza virus)
• Hantavirus pulmonary syndrome (Hantavirus)
• Hepatitis (Hepatitis virus)
• Chicken pox (Herpesvirus)
• Common cold (Picornavirus)
 Important Airborne Toxins
• Botulinal toxin
– Neurotoxin
– Lethal dose: 0.3 µg
• Staphylococcal enterotoxin
– Lethal dose ~ 25 µg
• LPS
– Endotoxin
– Sources: cotton mills, haystacks, sewage treatment
plants, solid waste handling facilities, swine
confinement buildings, poultry houses, homes & office
buildings.
– 10 ng can produce a toxic effect.
Examples:

Legionella causes two types of disease

• Pneumonia 2-5% exposed get sick, mortality rate is 5-15%
• Pontiac fever 95% exposed get flu-like symptoms
Humans are an incidental host to Legionella and successful infection requires:
1) A natural reservoir – any aquatic environment
2) Amplifying factors – warm water
3) Aerosolization – production of droplets of 5 um or less
4) A virulent strain of Legionella
5) Legionella must reach the alveolar region (lung)
6) Susceptibility of the host

Coccidiodes immitis – valley fever

Humans inhale spores released from the soil by wind, construction,

farming activities.

In the U.S. southwest, 30-40% of the population tests positive for

antibodies to Coccidiodes and 60% of the population has been infected
(many have no or mild symptoms).
 Aerosols
• Particles suspended in air.
• Respiratory intake and deposition in nasal and
bronchial airways.
• USEPA
– PM10 (diameter ≤ 10 µm)
– PM2.5 (diameter ≤ 2.5 µm)
– National Ambient Air Quality Standard (NAAQS)
• PM10 : 150 µg/m3 (1 day average), 50 µg/m3 (annual average)
• PM2.5 : 65 µg/m3 (1 day average), 15 µg/m3 (annual average)
Symptoms of particulate inhalation:
• Decreased pulmonary function
• Chronic coughs
• Bronchitis
• Asthmatic attacks
• Small particles are a greater concern
– Stay suspended longer and travel farther
– Move farther into respiratory system
– Viruses vs. bacteria

Particle Diameter Particle type Rate of fall in air

(mm) (cm /s)
1 Sand 7880
0.1 Silt 79
0.001 Clay 7.9 x 10-5
0.002 Clostridial spore 0.016
 London, 1952

Level of smoke and sulfur dioxide aerosols

associated with coal combustion was
elevated
4000 deaths in 10 days
(cardiovascular and lung disorders)

Gobi Desert, 1998-2001

Intense dust storms elevated aerosol

levels in western North America, which
is several thousands miles away
Biological Warfare – there is a long history

Romans threw dead animals into enemy water supplies

Tartars catapulted dead bodies infected with bubonic plague into Kaffa

British gave blankets used on smallpox victims to Native Americans

WWII: Japan sprayed bubonic plague over parts of China

1943: Great Britain tested anthrax spore viability and spread spores over
Gruinard Island

1951: U.S. sprayed Serratia marcesens over San Franciso

1966: U.S. released Bacillus subtilis into New York subway system

1984: The Dalles, OR. Salmonella and salad bars

 What are bioaerosols?
• Particles that range from 0.02 to 100 µm.
• Bioaerosols can be liquid or solid or both (e.g., a liquid droplet
containing a microbe).

Bacteria
Fungi
Protozoa

Viruses

0.01 um 0.1 um 1 um 10 um 100 um

Nuclei mode Accumulation Coarse mode
mode

fine particles coarse particles

 Aerobiological pathway

1. Launching (point source vs area source)

2. Transport (distance vs. survival)

3. Deposition (gravity and weather)

A sneeze vs. Release of wheat rust fungi

from a field
1. Launching

Total global emission of bacteria containing particles to

the atmosphere is estimated at 7.6 x 1023 to 3.5 x 1024.

Point source: isolated and well-defined site of

launching

-- Instantaneous point source: sneeze

-- continuous point source: biosolid pile

Linear source and area source involve larger, less

well-defined area

-- instantaneous linear source: passing aircraft

releasing biological warfare agent

-- continuous area source: release of bioaerosols

from a large field that has received an application
of biosolids or animal manures
 2. Transport (distance vs. survival)

distance time
Submicroscale up to 100 m 10 min
Microscale 100 m to 1 km 10 min to 1 hr
Mesoscale up to 100 km days
Macroscale larger longer
An example of mesoscale transport:

Aphthovirus (Picornaviridae)
The virus can persist outside the host for a month or more in
damp soil, aided by cold temperatures. Plumes of virus
contained within droplets, excreted at very high concentrations
from symptomatic animals, are dispersed by wind over long
distances (up to 60 km over land and 250 km over water).

A 1967 hoof and mouth disease outbreak in England impacted

2300 farms and 450,000 animals

It is estimated that bioaersols traveled > 60 km

A 2001 hoof and mouth disease outbreak in England caused >

760,000 animals to be slaughtered
 3. Deposition (gravity and weather)

a. gravitational settling
b. downward molecular diffusion
c. rainfall/electrostatic deposition

Stokes Law

ν (cm/sec) = ρ d2 g
18 η

Where: ν = terminal velocity

ρ = particle density (g/cm3)
d = particle diameter (cm)
g = acceleration due to gravity (cm/sec2)
η = viscosity of air (g/cm-sec)
Example: Consider a small particle such as an airborne microbe of 2
µm in diameter. How long does it take to fall to the ground from a
height of 3.05 m in the absence of any wind?

g = 981 cm/sec2
ρ = 1.3 g/cm3
η = 1.8 x 10-4 g/cm-sec
Downward molecular diffusion

Natural air currents and eddies enhance

the downward movement of particles and
increase the rate of deposition predicted
from gravitational settling by Stokes Law

Surface Impaction

Airborne particles can collide with numerous objects including other

small particles, trees, buildings, etc. After a collision, the particle can
stick or bounce off.
Rain and Electrostatic Deposition
Both processes condense a small particle into a larger one either associated
with a raindrop or associated electrostatically with another particle.

Example: Consider a microbe associated with a raindrop of 0.1 cm in

diameter. How long does it take to fall to the ground from a height of 3.05 m
in the absence of any wind?

g = 981 cm/sec2
ρ = 1.0 g/cm3
η = 1.8 x 10-4 g/cm-sec

Compare to earlier example

Microbial survival in aerosols

Depends on:
1. Type of microorganism Inactivation: Xt = X0-kt
2. Relative humidity
Where: X = number of microbes
3. Temperature k = inactivation constant
4. Radiation t = time

5. O2, OAF and Ions

Gram – microbes survive better at low humidity
Gram + microbes survive better at high humidity
Enveloped virus survive better at low humidity
Nonenveloped virus survive better at high humidity

Increasing temperature decreases survival

Open air factor: environmental factors produced when O3 reacts with

hydrocarbons
Extramural Aeromicrobiology

Antrax
1997: accidental release of bioaerosol from military microbiological
facility causing at 66 death in USSR

The Spanish Influenza Pandemic

1918: affecting 1/3 of world population with 50-100 millions death
Caused by H1N1

Microbes in the clouds:

Some microbes are ice nucleators that catalyzing ice formation
and may play a role in the formation and precipitation within
clouds.
Cloud environmental factors that can adversely influence microbes
Closed Environments

• Newer workplace buildings rely on air circulation systems instead

of windows to provide fresh air.

• Similarly, new home construction allows less air loss to save on

heating and air conditioning

• Extended space travel will require recirculation of all air and

water supplies.

• All of these situations lead to increased potential for LPS and

microbes to be recirculated throughout these spaces.
 Bioaerosol control
1. Ventilation – open the windows

2. Filtration – air circulation filters are rated on the dust spot

percentage which is an index of the size of particle the filter
removes. Typical rating for a building is 30 to 50%. This does
not remove particles such as LPS or virus. HEPA filters can
remove virus particles but are expensive so are not used in
buildings. This is the type of filter found in laminar flow hoods.

3. Biocides – thermal or UV treatment

4. Isolation/containment – may be used in a hospital setting

- Positive pressure
- Negative pressure
Biosafety levels

Level 1 – standard microbiology lab safety practices

Level 2 – special training

use of biological safety class II cabinets

Level 3 – restricted access labs

use of biological safety class III cabinets

Level 4 – restricted access to isolated labs

use of biological safety class III cabinets or biosafety
suits in combination with class II cabinets

Microbes identified as health risks are classified as class II, class

III, and class IV agents and must be worked with accordingly.
Some class II examples:
Class II Bacterial Campylobacter spp.
Clostridium spp.
E. Coli spp.

Fungal Penicillium spp.

Cryptococcus spp.
Microsporum spp.
Protozoal Cryptosporidium spp.
Giardia spp.
Encephalitozoon spp.
Enterocytozoon spp.
Entamoeba spp.
Leishmania spp.
Plasmodium spp.
Schistosoma spp.
Trypanosoma spp.
Viral Adenoviruses
Cowpox virus
Coxsackie A and B viruses
Hepatitis viruses A, B, C, D, & E
Epstein-Barr virus
Influenza viruses
Rhinoviruses
Some class III and IV examples:

Class III Bacterial Brucella spp.

Mycobacterium bovis
Mycobacterium tuberculosis
Rickettsia spp.
Yersinia pestis
Fungal Coccidioides immitis
Histoplasma capsulatum
Protozoal None
Viral Dengue virus
Monkey pox virus
Yellow fever virus

Class IV Bacterial None

Fungal None
Protozoal None
Viral Hemmorrhagic fever agents
Ebola fever virus
Marburg virus
HEPA filtered air Class II Hood
Room air
Contaminated air
A
B

C
Class III Hood B

A D

Front view

Room air
Contaminated air
HEPA filtered air