Inactivation of Bacterial Bioterrorism Agents in Water:
Summary of Seven Studies
INTRODUCTION
In the United States, chlorine and monochloramine are the primary chemical disinfectants used
to inactivate microbes in drinking water distribution systems. Although many microbes are
inactivated by common water treatments, some are more resistant. Conditions for inactivating
many waterborne disease-causing microbes have been
established, but there are only limited data on inactivating
bacterial bioterrorism agents.
U.S. EPA's Homeland Security Research Program
(HSRP) develops products based on scientific
research and technology evaluations. Our products
and expertise are widely used in preventing,
preparing for, and recovering from public health and
environmental emergencies that arise from terrorist
attacks. Our research and products address
biological, radiological, or chemical contaminants that
could affect indoor areas, outdoor areas, or water
infrastructure. HSRP provides these products,
technical assistance, and expertise to support EPA's
roles and responsibilities under the National
Response Framework, statutory requirements, and
Homeland Security Presidential Directives.
U.S. EPA and the Centers for Disease Control and
Prevention (CDC) have conducted seven laboratory-based
inactivation studies in water using non-disease causing
surrogates for Bacillus anthracis and microbes identified
as potential bioterrorism agents. One of the studies also
examined the conditions under which boiling water could
inactivate microscopic resistant structures (spores) formed
by surrogates.
A number of factors influence the effectiveness of
chemical disinfectants in drinking water treatment
systems, including:
• the type and quantity of microbes present
• whether the microbes form spores or exist primarily as vegetative cells
• the type of disinfectant and its concentration
• the amount of time the disinfectant is in contact with the microbes
• water temperature
• water acidity or alkalinity (pH)
• the type and quantity of organic and inorganic particles in the water
. water flow and pipe materials
Different species and strains of bacteria, whether bioterrorism agents or not, can have different
degrees of resistance to disinfectants. If nutrients are available prior to inactivation treatments,
this can increase the resistance of some species to chemical disinfection. Clumping or
attachment to floating organic materials can increase resistance. Some strains produce material
outside their cell wall. This extracellular material can permit attachment to other organisms or
surfaces and help form biologically active layers (biofilms), which are generally more resistant to
chemical disinfection than free floating (planktonic) cells.
Conditions in the inactivation studies were controlled. The data obtained, while suggestive,
cannot be directly applied to water distribution systems without factoring in circumstances that
will affect the how long a specific disinfectant and its residuals will be in contact with the
microorganisms.
May 2012
EPA/600/R-12/521
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OVERVIEW OF STUDIES ON THE INACTIVATION OF BIOTERRORISM
AGENTS AND SURROGATES IN WATER
The agents investigated can cause diseases in humans or animals from one or more of these
exposure routes [see References, Inactivation Studies 1, 2, 3, 5, 6, 7]:
• Direct contact with mucous membranes or broken skin
• Ingestion of contaminated food or water
• Inhalation of contaminated aerosols, dust, or particles
All of the agents have been investigated as possible biological weapons in the state-sponsored
research of one or more countries. Some have been used as biological weapons. All are
considered inhalation threats. In addition, four spore-forming surrogates, which are used by
many researchers in place of the virulent Bacillus anthracis Ames, were investigated: B.
anthracis Sterne; B. cereus; B. globigii; B. thuringiensis subsp. israelensis [see References,
Inactivation Studies 3, 4, 5, 6, 7].
The two common inactivation methods in water (chlorine and monochloramine) were tested on
26 strains of the following seven bioterrorism agents:
Bacterial Agent
Bacillus anthracis a
Brucella melitensis
Brucella suis
Burkholderia mallei
Burkholderia pseudo-mallei
Francisella tularensis
Yersinia pestis
Disease Caused
anthrax
brucellosis
brucellosis
glanders
melioidosis
tularemia
plague
CDC Category
A
B
B
B
B
A
A
On Select Agents List
HHS/APHIS
HHS/APHIS
HHS/APHIS
HHS/APHIS
HHS/APHIS
HHS
HHS
a forms spores
Bacterial agents are classified, transported, handled, and tested
according to definitions and regulations
CDC Category - The Centers for Disease Control and Prevention (CDC), U.S. Department of Health
and Human Services, categorize bioterrorism agents and diseases based on the degree to which they
pose a national security risk. Highest-priority agents (Category A) can be easily spread in the
environment or from person-to-person, result in high death rates, would potentially cause panic and
social disruption, and require special public health preparedness. Category B agents are moderately
easy to spread in the environment, result in a moderate number of illnesses and low death rates, and
require some changes to public health preparedness.
On Select Agents List - The Select Agents and Toxins List is defined by the Centers for Disease
Control and Prevention, U.S. Department of Health and Human Services (HHS) and the Animal and
Plant Health Inspection Service, U.S. Department of Agriculture (APHIS) and lists biological agents or
toxins deemed a threat to the public, animal or plant health, or to animal or plant products. There are
regulations on handling, transporting, and using select agents for research and testing.
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Table 1 summarizes the bacterial bioterrorism agent strains and treatments with the reference
numbers of the studies.
Table 1 Inactivation Studies of Bioterrorism Agents
Agents
Bacillus anthracis a
Brucella melitensis
Brucella suis
Burkholderia mallei
Burkholderia
pseudomallei
Francisella tularensis
Yersinia pestis
Strains Tested
Ames
ATCC 23456 C[NCTC1 0094] d
EAM562
M0562
M-9
M-13
ATCC 11 668
[NCTC11642]
ATCC 23343
[NCTC 12939]
ATCC 1688
[NCTC 1688]
AU 631 (soil)
CA 650 [ART]6
CA 652 [ART]
KC872
SC 763 [ART]
SC 764 [764]
TH 694 (water)
subsp. holarctica KY99-3387 (type B)
subsp. holarctica LVS (type B) (vaccine)
subsp. holarctica NY98
subsp. holarctica OR96-0246 (type B)
subsp. tularensis Schu S4 (A1)
subsp. tularensis WY96-3418 (A2)
subsp. tularensis MAOO-2987 (A1)
subsp. tularensis NM99-1823 (A2)
A1122 (low virulence)
Harbin
Inactivation Method
and Reference Number of Study
Chlorine
6b
6
6
6
2,6
1,6
6
Monochloramine
5
5
2
2,5
5
5
B. anthracis Ames and its surrogates form spores, the other bacteria in the inactivation studies live in vegetative
cell stages, which are less resistant to inactivation; B. anthracis surrogates are itemized in Table 2.
Study [6] has the original experimental data derived at 25 °C, which is cited in studies [3] and [7] with the temperature
listed as 23 °C.
c ATCC and associated number are registered or nonregistered trademarks of the American Type Culture Collection,
Manassas, Virginia, USA.
[NCTC] - the strain currently is listed by the National Collection of Type Cultures, Health Protection Agency, Salisbury, UK,
but no longer listed in the ATCC.
e [ART] - is from the "Antimicrobials Resistance Team, CDC" (see reference [2])
Table 2 summarizes the surrogates for B. anthracis and treatments with the reference numbers
of the studies.
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Table 2 Inactivation Studies of Surrogates for Bacillus anthracis Ames
Bacteria Used as Surrogates for
Bacillus anthracis Ames
Bacillus anthracis Sterne 34F2 [NCTC 8234] a
Bac/7/os cereus ATCC 7039 b
Bacillus cereus (commercial)
Bacillus globigii (Dugway) c
Bacillus thuringiensis subsp. israelensis ATCC 35646
Inactivation Method and Reference Number
Boiling in
Tap Water
4
4
4
4
Free Available
Chlorine
3,6
3
7
3
Monochloramine
5
[NCTC] — the strain currently is listed by the National Collection of Type Cultures, Health Protection Agency, Salisbury, UK.
ATCC and associated number are registered or nonregistered trademarks of the American Type Culture Collection,
Manassas, Virginia, USA.
c Strain from the U.S. Army Dugway Proving Ground, Utah
RESULTS FROM THE CHEMICAL INACTIVATION STUDIES [1,2, 3, 5, 6, 7]
A chemical disinfectant must be in contact with organisms for the length of time needed to
inactivate them and keep them inactivated. The condition needed for inactivation is expressed
as the Ct value (mg*min/L).
The Ct value is derived from experimental data and represents the disinfectant concentration
(C, in milligrams per liter) multiplied by the contact time (t, in minutes). Ct values are used to
establish the required conditions needed to achieve the desired amount of inactivation (Iog10
reduction) for a particular microorganism under a specific temperature and pH. Different
concentration and contact time combinations can result in the same Ct value.
Conditions that have an effect on the values include:
• the number and characteristics of the microbes,
• water temperature and pH
• quantity of suspended or attached particles
• disinfectant concentration
• water treatment system infrastructure (pipe materials, pipe loop designs, and age).
The typical conditions for chlorine (free available chlorine or FAC) and monochloramine - At the time
the inactivation studies were conducted, the conditions for chlorine treatment a at surveyed water treatment
plants were a median FAC of 1.1 mg/L and a median contact time from the treatment facility to the drinking
water customer of 45 minutes. For monochloramine treatment , the conditions were a median target
concentration of 2 mg/L and a median contact time from facility to customer of 45 minutes.
Water Quality Disinfection Committee. 1992. Survey of water utility disinfection practices. J. Am. Water Works Assoc. 84(9): 1-128.
Seidel, C.J., McGuire, M.J., Summers. R.S., and Via, S. 2005. Have utilities switched to chloramines? J. Am. Water Works Assoc. 97(10): 87-97.
Table 3 summarizes either the highest Ct value (mg*min/L) for a species that had multiple
strains tested in one or more studies ( = highest) or the only Ct value (mg*min/L) observed for a
species and strain in only one study (= single). These Ct values are indicative of the efficacy of
the disinfectants on a particular species and strain under particular temperatures and pH values.
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Table 3 Summary of Bioterrorism Agent and Surrogate Inactivation Results Expressed as Ct
Values
Microorganisms Tested
Bacillus anthracis
Bacillus cereus
Bacillus globigii
Bacillus thuringiensis subsp. israelensis
Brucella melitensis
Brucella suis
Burkholderia mallei
Burkholderia pseudomallei
Francisella tularensis subsp. holarctica
Yersinia pestis
Strains With
Highest or Single
Ct (mg*min/L) Values
Ames
Sterne 34F2
ATCC 7039
(Dugway)
ATCC 35646
ATCC 23456
EAM562
M0562
M-9
M-13
CA 652 (mucoid clinical)
TH 694 (water)
KY99-3387 (type B)
NY98 (type B)
A1122
Temperature
°C
5
15
25
5
15
25
5
23
5
23
5
23
5
15
25
5
25
5
15
25
5
15
25
5
25
5
25
5
25
5
25
5
15
25
5
15
25
Highest or Single
Ct (mg*min/L) Values
for 3 logio Inactivation
Chlorine
pH7
339
—
102
271
—
86
175
62
446
136
344
99
0.5
—
0.2
0.4
0.2
0.2
—
0.2
0.2
0.2
3.7
1.3
18.3
1.0
10.3 a
—
3.9
0.7
—
0.6
Mono-
chloramine
pH8
6,813
1,691
1,204
15,164
3,925
1,847
579.5
223.9
116.6
156.8
120.4
56.1
194.1
102.4
64.6
477
113
116.0
64.8
37.1
115.6
86.4
33.1
— not tested under these conditions a extrapolated, see reference |
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Summary of the Major Conclusions from Studies on Chemical Inactivation of
Bioterrorism Agents [1,2,5,6]
Agent
Under Typical Free Available Chlorine
Conditionsa
Under Typical Monochloramine
Conditions b
[6] 6. anthracis would not be inactivated by a
2 logio reduction
[5] depending on the temperature, Ct values for 6.
anthracis Sterne [surrogate] were 1.5 to 3 times
greater than those of Ames [virulent strain]
Bacillus anthracis
[6] "The Ames strain was slightly less
susceptible to the chlorine than the Sterne
strain, requiring more than 2 h for a 2 logio
reduction when exposed to 0.8 mg/L FAC at
25 °C, whereas the Sterne strain underwent a
> 4 logio reduction in counts after 2 h under
similar conditions."
[5] 6. anthracis Ames spores cannot be reduced
by 2 or 3 logio "regardless of temperature and
would require hours or days of disinfectant
exposure"
Brucella spp.
[6] 6. suis EAM562 and B. melitensis ATCC
23456 would be reduced by 3 logio if "pH and
temperature were similar to those in the
present study" {5 and 25 °C (41 and 77 °F) and
PH7}
[5] 6. suis MO 562 and a melitensis ATCC 23456
"would require a longer contact time or higher
disinfectant concentration for a 2 logio reduction"
Burkholderia
mallei
[6] a mallei M-9 and M-13 would be reduced
by 3 logio if pH and temperature were similar to
those in the present study" {5 and 25 °C
(41 and 77 °F) and pH 7}
[5] a mallei M-9...."demonstrated a 2 logio
inactivation" at a Ct value of 52.5 at 25 °C
Burkholderia
pseudomallei
{Strains tested in [2] B. pseudomallei AU 631; TH
694; SC 763; SC 764; ATCC 11668; ATCC 23343;
CA650;CA652}
[2] the planktonic populations of tested strains
of a pseudomallei could be reduced by
4 logio (in less than 10 minutes)
[2] For a 3 logio reduction, an 18-fold difference
was seen between the Ct values of the most
and least resistant strains (Ct values ranged
from 0.2 to 3.7 mg*min/L at pH 7 and 5 °C
(41 °F)
[2] The relative sensitivity to chlorine was
determined to be independent of a strain's
environmental, clinical, or culture collection
origins
[2] The relative amount of extracellular material
produced by a strain increased its tolerance to
chlorine
[6] a pseudomallei ATCC 1688 would be
reduced by 3 logio "pH and temperature were
similar to those in the present study" {5 and 25
°C (41 and 77 °F) and pH 7}
{Strains tested in [2] for monochloramine were B.
pseudomallei Ml 631; TH 694; SC 763; SC 764; ATCC
11668; ATCC 23343; CA 650; CA 652}
[2] the planktonic populations of tested strains of a
pseudomallei could be not reduced by
4 logio
[2] "Ct values were less variable than FAC Ct
values, differing by a factor of 2.5 between tested
strains"
[2] Ct values were independent of the amount of
extracellular material produced by each strain
[5] a pseudomallei KC 872 would be reduced by
2 logio
Median FAC of 1.1 mg/L and a median contact time from the treatment facility to the drinking water customer of 45 minutes
Median target concentration was 2 mg/L and a median contact time from the treatment facility to the drinking water customer of
45 minutes
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Summary of the Major Conclusions from Studies on Chemical Inactivation of
Bioterrorism Agents [1,2,5,6] ....continued
Agent
Under Typical Free Available Chlorine
Conditionsa
Under Typical Monochloramine
Conditions b
Francisella
tularensis
{Strains tested in [1] for FAC were, F. tularensis
subsp. holarctica KY99-3387 (type B), LVS (type
B), and OR96-0246 (type B) and F. tularensis
subsp. tularensis MAOO-2987 (A1), NM99-1823
(A2), Schu S4 (A1), and WY96-3418 (A2)}
[1] A 4 logio reduction of viable F. tularensis
counts occurred most rapidly at 25 °C (77 °F)
and pH 7; there was no significant difference
between the Ct values for all tested strains
under these conditions; disinfection occurred
most slowly at pH 8 and 5 °C (41 °F)
[1] For all conditions other than pH 7 and 25
°C (77 °F), the live vaccine strain, F.
tularensis subsp. holarctica LVS (type B),
was more sensitive to chlorine than the other
strains.
[1] The study recommended, when possible,
using Type B strains with full virulence, rather
than the live vaccine strain to avoid
underestimating Ct values needed for
disinfection
[1] The most favorable temperature 25 °C
(77 °F) and pH 7 combination would reduce the
planktonic population of the most sensitive
strain by 4 logio in less than one minute
[1] The least favorable temperature (5 °C (41
°F) and pH 8 would require up to 1.7 hours to
reduce the planktonic population of the most
tolerant strain by 4 logio
[[6] F. tularensis subsp. holarctica LVS (type
B) and F. tularensis subsp. holarctica NY 98
(type B) would be reduced by 3 logio if "pH and
temperature were similar to those in the
present study" {5 and 25 °C (41 and 77 °F) and
PH7}
[5] F. tularensis subsp. holarctica LVS (type B)
and F. tularensis subsp. holarctica NY 98 (type
B) could be reduced by 3 logio if the temperature of
the water was 15 °C (59 °F) or higher and the pH
maintained at 8
Yersinia pestis
[6] Y. pestis A1122 and Harbin would be
reduced by 3 logio if "pH and temperature were
similar to those in the present study" {5 and 25
°C (41 and 77 °F) and pH 7}
[5] Y. pestis A1122 and Harbin could be reduced
by 3 logio under median conditions if the
temperature of the water was 15 °C (59 °F)or
higher the pH maintained at 8
Median FAC of 1.1 mg/L and a median contact time from the treatment facility to the drinking water customer of 45 minutes
Median target concentration was 2 mg/L and a median contact time from the treatment facility to the drinking water customer of
45 minutes
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Summary of the Major Conclusions from Studies on Chemical Inactivation of Surrogates
[3,5,6,7]
Surrogates for
Bacillus anthracis
Bacillus anthracis
Sterne
Bacillus cereus
Bacillus globigii
(Dugway)
Bacillus
thuringiensis
subs p. israelensis
Under Typical Free Available Chlorine
Conditions a
[3] Spores of 6. anthracis Sterne had "substantially
lower Ct values" than the spores of 6. thuringiensis
subsp. israelensis or of 6. anthracis Ames [6]
[3] Spores of 6. cereus had "substantially lower Ct
values" than the spores of 6. thuringiensis subsp.
israelensis or of 6. anthracis Ames [6]
[3] The "spores of 6. globigii were more resistant
than the spores of other Bacillus spp." in this and
other studies [6, 7]
[3] "Spores of 6. thuringiensis subsp. israelensis
would be an appropriate surrogate to use in place of
6. anthracis in chlorine inactivation studies"
Under Typical Monochloramine
Conditions b
[5] 6. anthracis Sterne spores cannot be
reduced by 2 or 3 logio "regardless of
temperature and would require hours or days
of disinfectant exposure"
Median FAC of 1.1 mg/L and a median contact time from the treatment facility to the drinking water customer of 45 minutes
Median target concentration was 2 mg/L and a median contact time from the treatment facility to the drinking water customer of
45 minutes
RESULTS FROM THE BOILING INACTIVATION STUDY [4]
Results of this study have implications for boil water advisories that are issued by public health
and other authorities. Many waterborne, disease-causing microbes can be inactivated at a
rolling boil held for one minute. However, even after five minutes of boiling in tap water in an
uncovered vessel, viable spores of all three tested species of Bacillus spp. were recovered.
Bac/7/us anthracis Sterne 34F2 [NCTC 8234]'
Bac/7/us cereus ATCC 7039
Bacillus cereus (commercial)
Bacillus thuringiensis subsp. israelensis ATCC 35646
3 Minutes Boiling
Covered
No Viable Spores Detected
5 Minutes Boiling
Uncovered
Viable Spores Detected
[NCTC] — the strain currently is listed by the National Collection of Type Cultures, Health Protection Agency, Salisbury, UK.
ATCC and associated number are registered or nonregistered trademarks of the American Type Culture Collection,
Manassas, Virginia, USA.
The average temperatures at boiling, immediately above the water surfaces, were 98.9 °C
(210.02 °F) for the covered vessels and 77.3 °C (171.14 °F) for the uncovered vessels. The
investigators cautioned that atmospheric pressure and altitude determine the temperature at
which water boils and this will affect inactivation conditions. Increasing altitude decreases
water's boiling point; increasing barometric pressure increases the boiling point of water.
Following the References, see Supplemental Tables 1s [free available chlorine] and 2s
rmonochloramine] for the Ct values for all species and strains tested in the seven studies.
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Summary Information on the Bacterial Bioterrorism Agents Used in the Inactivation Studies
Agent (Disease)
Transmission and Typical
Exposure Sources
Geographic Distribution and
Natural Hosts
Waterborne Threat?
Persistence
Bacillus anthracis
(Anthrax)
Direct person-to-person
transmission is rare from skin
infections and is not known
from inhalation; highly
infectious and has high
mortality rate; contact with
spore-contaminated soils or
infected animal by-products
such as bone, hair, hide, and
under-cooked meat are the
major sources of human
infection.
8. anthracis can be found worldwide, causing
anthrax primarily in grazing mammals such as
sheep, cattle, goats, camels, or wild animals
such as antelopes and deer. Human anthrax
cases are reported from Africa, Asia, Europe,
and the Americas, with only a few locations
free of any reported disease.
The NRTa considers 6.
anthracis a possible water
threat and cautions that re-
aerosol ization can occur
when using spore
contaminated water, for
example in fire fighting.
8. anthracis spores can remain
viable for decades under harsh
conditions in the environment and
inactivation of spores in biofilms
growing in the water treatment
system pipes is difficult.
Brucella spp.
(Brucellosis)
Direct person-to-person
transmission is rare, but has
been documented; worldwide, it
is the most common disease
transmissible to humans from
animals and is one of the most
common laboratory-acquired
diseases; ingesting
unpasteurized dairy products or
infected animal products is the
major exposure route.
Although largely eradicated in much of
Europe and North America, brucellosis
remains an important human and agricultural
health problem in parts of North Africa, the
Mediterranean, the Middle East, Asia, India,
and Central and South America; can infect a
variety of animals, including cattle, sheep,
goats, camels, pigs, dogs, reindeer, yaks and
many wild mammal species. Brucella spp. or
Brucella antibodies have been detected in
many marine mammals, including seals,
dolphins, porpoises, walruses, and whales.
The NRT considers Brucella
spp. a probable water threat
because the bacteria are
stable in water for 20 to 72
days.
Brucella suis and 8. melitensis can
persist in soil for up to 125 days.
Without exposure to sunlight, under
low to moderate temperatures (4 to
22 °C (39 to 72 °F)), 8. suis has
remained viable for at least 28 days
on aluminum, glass, and topsoil.
Burkholderia mallei
(Glanders)
Direct person-to-person
transmission is rare; infected
horses, with and without
symptoms, pose the greatest
risk for human exposure.
Many countries have eradicated naturally
occurring glanders, but it is still found in parts
of Africa, the Middle East, Central and South
America. Glanders is primarily a disease of
horses, mules, and donkeys, but can also be
found in other animals.
The NRT reports that 8.
mallei can survive in water at
room temperature in water
for one month.
8. mallei can survive in warm, moist
environments for a few months. It is
not believed to be persistent in soil.
NRT - National Response Team, see References: Bioterrorism Agents under "U.S. NRT"
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Summary Information on the Bacterial Bioterrorism Agents Used in the Inactivation Studies ....continued
Agent (Disease)
Transmission and Typical
Exposure Sources
Geographic Distribution and
Natural Hosts
Waterborne Threat?
Persistence
Burkholderia
pseudomallei
(Melioidosis)
Direct person-to-person
transmission is rare, but has
been documented; infectivity
when aerosolized in not known;
people with risk factors such as
diabetes or alcoholism are at
the greatest risk of contracting
severe forms of the disease;
melioidosis presents with a
wide variety of symptoms in
multiple body systems and
symptoms can take years to
first appear; it can be acute or
chronic.
8. pseudomallei is widely found in tropical and
subtropical climates, including Southeast
Asia, northern Australia, south Asia, and
China, as well as sporadically in parts of
Africa, Central and South America. Many
animal species, including sheep, goats,
horses, swine, cattle, kangaroos, camels,
dogs and cats, even some species of birds,
are susceptible to infection.
The NRTa reports that
8. pseudomallei \s known to
persist in water for over
three years.
8. pseudomallei is a resilient
microbe able to withstand hostile
environmental conditions and long
periods of nutritional deficiency. It
can persist in moist clay soils for up
to two years. It can survive in acidic
environments (pH 4.5) for 40 days.
Francisella
tularensis
(Tularemia)
Direct person-to-person
transmission is rare from skin
and not known from inhalation;
highly infectious when
aerosolized; can be acquired by
direct contact with infected
animals, animal bites, ingestion,
and inhalation, in addition to
being transmitted by arthropods
such as deerflies, mosquitos,
and ticks.
F. tularensis is found almost entirely in North
America and Eurasia. Many animals are
susceptible to tularemia, including rabbits and
hares, sheep, and many rodents.
The NRT considers water a
possible pathway for the
weaponized organism.
Natural outbreaks of F.
tularensis subsp. holarctica
in water have occurred.
F. tularensis can persist under cold,
moist conditions in hay, water,
decaying carcasses, and soil. Live
bacteria have been found in rabbit
meat after 3 years storage at -15 °C
(5 °F).
Yersinia pestis
(Plague)
Direct person-to-person
transmission of pneumonic
plague is possible; human
infection is usually from flea
bites; the plague has occurred
in pandemics as recently as the
1890's (started in China).
Y. pestis is found on all continents except
Australia and Antarctica. In the wild, Y. pestis
causes disease in over 200 species of
rodents as well as rabbits, cats, dogs, and
other animals, which can acquire the disease,
as well as carry infected fleas
The NRT considers it
possible that Y. pestis poses
a water threat and reports
that it has persisted in spring
water under laboratory
conditions for 160 days.
Under controlled conditions at 22 °C
(72 °F), Y. pestis has remained
viable for at least seven days on
aluminum and painted dry wall tape;
without sunlight, it can remain viable
and infectious under controlled
conditions in soil for up to 40 weeks.
NRT - National Response Team, see References: Bioterrorism Agents under "U.S. NRT"
10
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REFERENCES: Inactivation Studies
[1] O'Connell, H.A., Rose, L.J., Shams, A.M., Arduino, M.J., and Rice, E.W. 2011. Chlorine
disinfection of Francisella tularensis. Lett. Appl. Microbiol. 52(1): 84-86.
[2] O'Connell, H.A., Rose, L.J., Shams, A.M., Bradley, M., Arduino, M.J., and Rice, E.W. 2009.
Variability of Burkholderia pseudomallei strain sensitivities to chlorine disinfection. Appl.
Environ. Microbiol. 75(16): 5405-5409.
[3] Rice, E.W., Adcock, N.J., Sivaganesan, M., and Rose, L.J. 2005. Inactivation of spores of
Bacillus anthracis Sterne, Bacillus cereus, and Bacillus thuringiensis subsp. israelensis by
chlorination. Appl. Environ. Microbiol. 71(9): 5587-5589.
[4] Rice, E.W., Rose, L.J., Johnson, C.H., Boczek, L.A., Arduino, M.,J. and Reasoner, D.J.
2004. Boiling and Bacillus spores [letter]. Emerg. Infect. Dis. 10(10): 1887-1888.
[5] Rose, L.J., Rice, E.W., Hodges, L, Peterson, A., and Arduino, M.J. 2007. Monochloramine
inactivation of bacterial select agents. Appl. Environ. Microbiol. 73(10): 3437-3439.
[6] Rose, L.J., Rice, E.W., Jensen, B., Murga, R., Peterson, A., Donlan, R.M., and Arduino, M.J.
2005. Chlorine inactivation of bacterial bioterrorism agents. Appl. Environ. Microbiol. 71(1): 566-
568.
[7] Sivaganesan, M., Adcock, N.J., and Rice, E.W. 2006. Inactivation of Bacillus globigii by
chlorination: a hierarchical Bayesian model. J. Water Supply: Res. Technol.-AQUA. 55(1): 33-
43.
REFERENCES: Bioterrorism Agents
Dembeck, Z. F. 2007. Medical Aspects of Biological Warfare. Washington, D.C.: Borden
Institute: Walter Reed Army Medical Center.
Sinclair, R., Boone, S.A., Greenburg, D. Keim, P., and Gerba, C.P. 2008. Persistence of
category A select agents in the environment. Appl. Environ. Microbiol. 74(3):555-563.
Spickler, A. R. Technical Factsheets: Anthrax (2007); Brucellosis (2009); Glanders (2007);
Melioidosis (2007); Tularemia (2009); Plague (2009). Ames, Iowa: Center for Food Security and
Public Health, Iowa State University.
U.S. EPA. 2011. Results from Persistence Testing of Biological Agents Under Various
Conditions. Washington, D.C.: U.S. Environmental Protection Agency. EPA/600/S-11/005.
U.S. NRT. 2011. NRT Quick Reference Guides on Bacillus anthracis; Brucella Species;
Burkholderia mallei and Burkholderia pseudomallei; Francisella tularensis; Yersinia pestis.
CONTACT INFORMATION
For more information, visit the EPA Web site at www.epa.gov/nhsrc.
Technical Contact: Eugene W. Rice (rice.gene@epa.gov)
General Feedback/Questions: Kathy Nickel (nickel.kathy@epa.gov)
11
-------
Supplemental Table 1s. Free Available Chlorine Inactivation of Bacterial Strains at pH 7 or 8 and Temperatures
at 5, 23, or 25 °C (41, 73, or 77 °F)
Log™
Reduc-tion
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
4 Iog10
Bacterial Strains Tested
Bacillus anthracis Ames
Bacillus anthracis Ames
Bacillus anthracis Sterne 34F2
Bacillus anthracis Sterne 34F2
Bacillus anthracis Sterne 34F2
Bacillus anthracis Sterne 34F2
Bacillus anthracis Sterne 34F2
Bacillus cereus ATCC7039 (b)
Bacillus cereus ATCC 7039
Bacillus cereus ATCC 7039
Bacillus globigii (Dugway)
Bacillus globigii (Dugway)
Bacillus thuringiensis subsp. israelensis ATCC 35646
Bacillus thuringiensis subsp. israelensis ATCC 35646
Bacillus thuringiensis subsp. israelensis ATCC 35646
Temperature
25
25
25
25
23
23
23
23
23
23
23
23
23
23
23
•» At pH 7
Ct (mg*min/L) at
23 or 25 °C (73
or 77 °F)
79
102
60
86
45
68
90
41
62
82
108
136
66
99
132
Temperature
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
•» At pH 7
Ct (mg*min/L) at
5 °C (41 °F)
220
339
190
271
140
210
280
117
175
233
372
446
229
344
458
Temperature
25
25
25
25
23
23
23
23
23
23
23
23
23
23
23
•» At pH 8
Ct (mg*min/L) at 23
or 25 °C (73 or
77 °F)
, (a)
nt
nt
nt
nt
127
191
254
132
199
264
367
438
246
369
492
Temperature
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
•» At pH 8
Ct (mg*min/L) at 5
°C (41 °F)
nt
nt
nt
nt
319
478
637
340
510
680
943
1,144
481
721
961
Citations
Rose, L.J., et al. 2005. Appl. Environ. Microbiol . 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rice, E.W., et al. 2005. Appl. Environ. Microbiol .71(9): 5587-5589.
Rice, E.W., et al. 2005. Appl. Environ. Microbiol. 71(9): 5587-5589.
Rice, E.W., et al. 2005. Appl. Environ. Microbiol .71(9): 5587-5589.
Rice, E.W., et al. 2005. Appl. Environ. Microbiol. 71(9): 5587-5589.
Rice, E.W., et al. 2005. Appl. Environ. Microbiol. 71(9): 5587-5589.
Rice, E.W., et al. 2005. Appl. Environ. Microbiol. 71(9): 5587-5589.
Sivaganesan, M., etal. 2006. J. Water Supply: Res. Technol.-
AQUA. 55(1): 33-43.
Sivaganesan, M., etal. 2006. J. Water Supply: Res. Technol.-
AQUA. 55(1): 33-43.
Rice, E.W., et al. 2005. Appl. Environ. Microbiol .71(9): 5587-5589.
Rice, E.W., et al. 2005. Appl. Environ. Microbiol. 71(9): 5587-5589.
Rice, E.W., et al. 2005. Appl. Environ. Microbiol .71(9): 5587-5589.
Reference Number
[6]
[6]
[6]
[6]
[3]
[3]
[3]
[3]
[3]
[3]
[7]
[7]
[3]
[3]
[3]
Inactivation of Bacterial Bioterrorism Agents in Water
12
-------
Supplemental Table 1s. Free Available Chlorine Inactivation of Bacterial Strains at pH 7 or 8 and Temperatures
at 5, 23, or 25 °C (41, 73, or 77 °F)
Log™
Reduc-tion
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
Bacterial Strains Tested
Brucella melitensis ATCC 23456 [NCTC 10094] (c)
Brucella melitensis ATCC 23456 [NCTC 10094]
Brucella suis EAM562
Brucella suis EAM562
Burkholderia mallei M-13
Burkholderia mallei M-13
Burkholderia mallei M-9
Burkholderia mallei M-9
Burkholderia pseudomallei ATCC 1688 [NCTC 1688]
Burkholderia pseudomallei ATCC 1688 [NCTC 1688]
Burkholderia pseudomallei ATCC 1 1668 [NCTC 1 1642] -
source: clinical
Burkholderia pseudomallei ATCC 1 1668 [NCTC 1 1642] -
source: clinical
Burkholderia pseudomallei ATCC 1 1668 [NCTC 1 1642] -
source: clinical
Burkholderia pseudomallei ATCC 23343 [NCTC 12939] -
source: clinical
Burkholderia pseudomallei ATCC 23343 [NCTC 12939] -
source: clinical
Burkholderia pseudomallei ATCC 23343 [NCTC 12939] -
source: clinical
Burkholderia pseudomallei AU 631 - source: soil
Burkholderia pseudomallei AU 631 - source: soil
Burkholderia pseudomallei AU 631 - source: soil
CD
Z3
~O3
8.
CD
1—
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
•* AtpH7
Ct (mg*min/L) at
23 or 25 °C (73
or 77 °F)
0.1
0.2
0.1
0.2
0.1
0.2
0.1
0.2
0.4
0.6
0.3
0.5
0.8
0.7
0.9
1.1
0.1
0.1
0.1
CD
Z3
~O3
8.
CD
1—
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
* At pH 7
Ct (mg*min/L) at
5 °C (41 °F)
0.3
0.5
0.3
0.4
0.2
0.2
0.2
0.2
0.5
0.7
0.4
1.1
1.8
1.0
1.4
1.8
0.1
0.2
0.3
CD
Z3
"O3
8.
CD
1—
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
•* At pH 8
Ct (mg*min/L) at 23
or 25 °C (73 or
77 °F)
nt
nt
nt
nt
nt
nt
nt
nt
nt
nt
0.2
0.4
0.7
0.5
1.1
1.8
0.1
0.1
0.1
Temperature
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
•* At pH 8
Ct (mg*min/L) at 5
°C (41 °F)
nt
nt
nt
nt
nt
nt
nt
nt
nt
nt
0.7
1.3
1.9
0.9
1.9
2.8
0.2
0.3
0.4
Citations
Rose, L.J., et al. 2005. Appl. Environ. Microbiol . 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
Reference Number
[6]
[6]
[6]
[6]
[6]
[6]
[6]
[6]
[6]
[6]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
Inactivation of Bacterial Bioterrorism Agents in Water
13
-------
Supplemental Table 1s. Free Available Chlorine Inactivation of Bacterial Strains at pH 7 or 8 and Temperatures
at 5, 23, or 25 °C (41, 73, or 77 °F)
Log™
Reduc-tion
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
Bacterial Strains Tested
Burkholderia pseudomallei CA 650 - source: clinical,
transiently mucoid [ART]
Burkholderia pseudomallei CA 650 - source: clinical,
transiently mucoid [ART]
Burkholderia pseudomallei CA 650 - source: clinical,
transiently mucoid [ART]
Burkholderia pseudomallei CA 652 - source: clinical,
mucoid [ART]
Burkholderia pseudomallei CA 652 - source: clinical,
mucoid [ART]
Burkholderia pseudomallei CA 652 - source: clinical,
mucoid [ART]
Burkholderia pseudomallei SC 763 - source: clinical,
nonmucoid [ART]
Burkholderia pseudomallei SC 763 - source: clinical,
nonmucoid [ART]
Burkholderia pseudomallei SC 763 - source: clinical,
nonmucoid [ART]
Burkholderia pseudomallei SC 764 - source: clinical,
nonmucoid [ART]
Burkholderia pseudomallei SC 764 - source: clinical,
nonmucoid [ART]
Burkholderia pseudomallei SC 764 - source: clinical,
nonmucoid [ART]
Burkholderia pseudomallei TH 694 - source: water
Burkholderia pseudomallei TH 694 - source: water
Burkholderia pseudomallei TH 694 - source: water
CD
O3
8.
CD
75
25
25
75
25
75
25
75
25
75
25
75
25
25
25
•* AtpH7
Ct (mg*min/L) at
23 or 25 °C (73
or 77 °F)
0.6
1 0
1.5
0.8
1.3
1.7
0.2
0.3
0.4
0.1
0.1
0.1
0.1
0.1
0.2
CD
Z3
O3
8.
CD
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
* At pH 7
Ct (mg*min/L) at
5 °C (41 °F)
0.8
1 3
1.7
2.3
3.7
5.0
0.2
0.3
0.5
0.1
0.2
0.3
0.1
0.2
0.4
CD
Z3
O3
8.
CD
75
25
25
75
25
75
25
75
25
75
25
75
25
25
25
•* At pH 8
Ct (mg*min/L) at 23
or 25 °C (73 or
77 °F)
1.1
1 7
2.3
0.9
1.4
1.9
0.1
0.2
0.3
0.1
0.1
0.2
0.1
0.2
0.4
CD
Z3
O3
8.
CD
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
•* At pH 8
Ct (mg*min/L) at 5
°C (41 °F)
1.1
1 7
2.9
3.7
5.8
7.8
0.5
0.8
1.1
0.2
0.3
0.5
0.1
0.3
0.5
Citations
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16): 5405-
5409.
8
E
•z.
0>
o
£
"S
t£
[2]
[21
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
Inactivation of Bacterial Bioterrorism Agents in Water
14
-------
Supplemental Table 1s. Free Available Chlorine Inactivation of Bacterial Strains at pH 7 or 8 and Temperatures
at 5, 23, or 25 °C (41, 73, or 77 °F)
Log™
Reduc-tion
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
Bacterial Strains Tested
Francisella tularensis subsp. holarctica KY99-3387 (type
B)
Francisella tularensis subsp. holarctica KY99-3387 (type
B)
Francisella tularensis subsp. holarctica KY99-3387 (type
B)
Francisella tularensis subsp. holarctica LVS (type B)
Francisella tularensis subsp. holarctica LVS (type B)
Francisella tularensis subsp. holarctica LVS (type B)
Francisella tularensis subsp. holarctica LVS (type B)
Francisella tularensis subsp. holarctica LVS (type B)
Francisella tularensis subsp. tularensis MAOO-2987 (A1 )
Francisella tularensis subsp. tularensis MAOO-2987 (A1 )
Francisella tularensis subsp. tularensis MAOO-2987 (A1 )
Francisella tularensis subsp. tularensis NM99-1823(A2)
Francisella tularensis subsp. tularensis NM99-1823(A2)
Francisella tularensis subsp. tularensis NM99-1823(A2)
CD
Z3
"03
8.
CD
1—
25
25
25
25
25
25
25
25
25
25
25
25
25
25
•* AtpH7
Ct (mg*min/L) at
23 or 25 °C (73
or 77 °F)
0.8
1.0
1.3
0.6
1.0
0.7
1.0
1.2
0.9
1.3
1.6
0.4
0.5
0.7
CD
Z3
~O3
8.
CD
1—
5
5
5
5
5
5
5
5
5
5
5
5
5
5
* At pH 7
Ct (mg*min/L) at
5 °C (41 °F)
14.4
18.3
22.3
1.5
2.4
5.0
6.7
8.5
13.6
16.9
20.2
14.4
17.7
21.0
CD
Z3
"O3
8.
CD
1—
25
25
25
25
25
25
25
25
25
25
25
25
25
25
•* At pH 8
Ct (mg*min/L) at 23
or 25 °C (73 or
77 °F)
2.6
3.2
3.8
nt
nt
2.0
2.7
3.5
2.7
3.4
4.2
2.9
3.7
4.5
Temperature
5
5
5
5
5
5
5
5
5
5
5
5
5
5
•* At pH 8
Ct (mg*min/L) at 5
°C (41 °F)
33.9
43.7
53.5
nt
nt
15.9
20.1
24.3
64.1
83.8
103.4
45.4
60.5
75.7
Citations
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol . 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., et al. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
Reference Number
[1]
[1]
[1]
[6]
[6]
[1]
[1]
[1]
[1]
[1]
[1]
[1]
[1]
[1]
Inactivation of Bacterial Bioterrorism Agents in Water
15
-------
Supplemental Table 1s. Free Available Chlorine Inactivation of Bacterial Strains at pH 7 or 8 and Temperatures
at 5, 23, or 25 °C (41, 73, or 77 °F)
Log™
Reduc-tion
2 Iog10
3 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
Bacterial Strains Tested
Francisella tularensis subsp. holarctica NY98 (type B)
Francisella tularensis subsp. holarctica NY98 (type B)
Francisella tularensis subsp. holarctica OR96-0246 (type
B)
Francisella tularensis subsp. holarctica OR96-0246 (type
B)
Francisella tularensis subsp. holarctica OR96-0246 (type
B)
Francisella tularensis subsp. tularensis Schu S4(A1)
Francisella tularensis subsp. tularensis Schu S4(A1)
Francisella tularensis subsp. tularensis Schu S4(A1)
Francisella tularensis subsp. tularensis WY96-3418(A2)
Francisella tularensis subsp. tularensis WY96-3418(A2)
Francisella tularensis subsp. tularensis WY96-3418(A2)
Yersinia pestis A1122
Yersinia pestis A1 122
Yersinia pestis Harbin
Yersinia pestis Harbin
Temperature
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
•» At pH 7
Ct (mg*min/L) at
23 or 25 °C (73
or 77 °F)
2.0
3.9
0.9
1.2
1.5
0.9
1.3
1.7
0.8
1.3
1.6
0.4
0.6
0.03
0.04
Temperature
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
•» At pH 7
Ct (mg*min/L) at
5 °C (41 °F)
7.8
10.3 (e)
9.3
12.9
16.5
13.4
16.8
20.3
14.2
17.4
20.8
0.5
0.7
0.03
0.04
Temperature
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
•» At pH 8
Ct (mg*min/L) at 23
or 25 °C (73 or
77 °F)
nt
nt
2.7
3.7
4.6
3.7
4.5
5.2
3.3
4.1
5.0
nt
nt
nt
nt
Temperature
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
•» At pH 8
Ct (mg*min/L) at 5
°C (41 °F)
nt
nt
47.1
59.0
70.8
47.4
62.3
77.2
46.8
61.7
76.2
nt
nt
nt
nt
Citations
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
O'Connell, H.A., etal. 2011. Lett. Appl. Microbiol. 52(1): 84-86
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Rose, L.J., et al. 2005. Appl. Environ. Microbiol. 71(1): 566-568.
Reference Number
[6]
[6]
[1]
[1]
[1]
[1]
[1]
[1]
[1]
[1]
[1]
[6]
[6]
[6]
[6]
KEY nt = not tested ATCC and the associated number are registered or nonregistered trademarks of the American Type Culture Collection, Manassas, Virginia, USA [NCTC] - the strain currently is listed by National Collection of Type Cultures, Health
Protection Agency, Salisbury, UK, but no longer listed in the American Type Culture Collection ART — source is the "Antimicrobials Resistance Team, CDC" (see reference 2) extrapolated number (see reference 6)
Inactivation of Bacterial Bioterrorism Agents in Water
16
-------
Supplemental Table 2s. Monochloramine Inactivation of Bacterial Strains at pH 8 and Temperatures
at 5, 15, or 25 °C (41, 59, or 77 °F)
Log™
Reduc-
tion
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
Bacterial Strains Tested
Bacillus anthracis Ames
Bacillus anthracis Ames
Bacillus anthracis Sterne 34F2
Bacillus anthracis Sterne 34F2
Brucella melitensis ATCC 23456 (a) [NCTC 10094] (b)
Brucella melitensis ATCC 23456 [NCTC 10094]
Brucella suis M0562
Brucella suis M0562
Burkholderia mallei M-9
Burkholderia mallei M-9
Burkholderia pseudo-mallei ATCC 1 1668 [NCTC 1 1642] - source:
clinical
Burkholderia pseudomallei ATCC 1 1668 [NCTC 1 1642] - source:
clinical
Burkholderia pseudomallei ATCC 1 1668 [NCTC 1 1642] - source:
clinical
Burkholderia pseudomallei ATCC 23343 [NCTC 12939] - source:
clinical
Burkholderia pseudomallei ATCC 23343 [NCTC 12939] - source:
clinical
Burkholderia pseudomallei ATCC 23343 [NCTC 12939] - source:
clinical
Temperature
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
•» At pH 8
Ct (mg*min/L) at 25
°C (77 °F)
785
1,204
1,442
1,847
104.4
116.6
47.8
56.1
52.5
64.6
43
49
54
49
73
97
Temperature
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
•» At pH 8
Ct (mg*min/L) at 15
°C (59 °F)
1,072
1,691
2,793
3,925
204.0
223.9
99.8
120.4
89.4
102.4
„(«>
nt
nt
nt
nt
nt
Temperature
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
•» At pH 8
Ct (mg*min/L) at 5
°C (41 °F)
3,499
6,813
10,532
15,164
501.8
579.5
134.3
156.8
158.6
194.1
204
238
262
190
226
251
Citations
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
Reference
Number
[5]
[5]
[5]
[5]
[5]
[5]
[5]
[5]
[5]
[5]
[2]
[2]
[2]
[2]
[2]
[2]
Inactivation of Bacterial Bioterrorism Agents in Water
17
-------
Supplemental Table 2s. Monochloramine Inactivation of Bacterial Strains at pH 8 and Temperatures
at 5, 15, or 25 °C (41, 59, or 77 °F)
Log™
Reduc-
tion
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
4 Iog10
Bacterial Strains Tested
Burkholderia pseudomallei AU 631 - source: soil
Burkholderia pseudomallei AU 631 - source: soil
Burkholderia pseudomallei AU 631 - source: soil
Burkholderia pseudomallei CA 650 - source: clinical, transiently mucoid
[ART](d)
Burkholderia pseudomallei CA 650 - source: clinical, transiently mucoid
[ART]
Burkholderia pseudomallei CA 650 - source: clinical, transiently mucoid
[ART]
Burkholderia pseudomallei CA 652 - source: clinical, mucoid [ART]
Burkholderia pseudomallei CA 652 - source: clinical, mucoid [ART]
Burkholderia pseudomallei CA 652 - source: clinical, mucoid [ART]
Burkholderia pseudomallei KG 872
Burkholderia pseudomallei KG 872
Burkholderia pseudomallei SC 763 - source: clinical, nonmucoid [ART]
Burkholderia pseudomallei SC 763 - source: clinical, nonmucoid [ART]
Burkholderia pseudomallei SC 763 - source: clinical, nonmucoid [ART]
Burkholderia pseudomallei SC 764 - source: clinical, nonmucoid [ART]
Burkholderia pseudomallei SC 764 - source: clinical, nonmucoid [ART]
Burkholderia pseudomallei SC 764 - source: clinical, nonmucoid [ART]
Temperature
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
•» At pH 8
Ct (mg*min/L) at 25
°C (77 °F)
42
49
55
50
68
86
70
88
99
38.8
45.9
53
60
68
48
56
65
Temperature
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
•* At pH 8
Ct (mg*min/L) at 15
°C (59 °F)
nt
nt
nt
nt
nt
nt
nt
nt
nt
87.6
103.9
nt
nt
nt
nt
nt
nt
Temperature
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
•» At pH 8
Ct (mg*min/L) at 5
°C (41 °F)
240
266
291
138
202
266
234
281
328
116.7
156.1
302
382
462
266
288
310
Citations
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
Reference
Number
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[2]
[5]
[5]
[2]
[2]
[2]
[2]
[2]
[2]
Inactivation of Bacterial Bioterrorism Agents in Water
18
-------
Supplemental Table 2s. Monochloramine Inactivation of Bacterial Strains at pH 8 and Temperatures
at 5, 15, or 25 °C (41, 59, or 77 °F)
Log™
Reduc-
tion
2 Iog10
3 Iog10
4 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
2 Iog10
3 Iog10
Bacterial Strains Tested
Burkholderia pseudomallei TH 694 - source: water
Burkholderia pseudomallei TH 694 - source: water
Burkholderia pseudomallei TH 694 - source: water
Francisella tularensis subsp. holarctica LVS (type B)
Francisella tularensis subsp. holarctica LVS (type B)
Francisella tularensis subsp. holarctica NY98 (type B)
Francisella tularensis subsp. holarctica NY98 (type B)
Yersinia pestis A1 1 22
Yersinia pestis A1122
Yersinia pestis Harbin
Yersinia pestis Harbin
Temperature
25
25
25
25
25
25
25
25
25
25
25
-» At pH 8
Ct (mg*min/L) at 25
°C (77 °F)
99
113
127
26.3
30.4
31.3
37.1
27.6
33.1
21.9
25.0
Temperature
15
15
15
15
15
15
15
15
15
15
15
•» At pH 8
Ct (mg*min/L) at 15
°C (59 °F)
nt
nt
nt
61.2
71.1
48.7
64.8
71.4
86.4
33.5
40.8
Temperature
5
5
5
5
5
5
5
5
5
5
5
•* At pH 8
Ct (mg*min/L) at 5
°C (41 °F)
404
477
550
76.0
97.9
84.0
116.0
92.0
115.6
80.7
91.4
Citations
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
O'Connell, H.A., et al. 2009. Appl. Environ. Microbiol. 75(16):
5405-5409.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Rose, L.J., et al. 2007. Appl. Environ. Microbiol. 73(10): 3437-
3439.
Reference
Number
[2]
[2]
[2]
[5]
[5]
[5]
[5]
[5]
[5]
[5]
[5]
KEY ATCC and the associated number are registered or nonregistered trademarks of the American Type Culture Collection, Manassas, Virginia, USA [NCTC] - the strain currently is listed by National Collection of Type Cultures, Health
Protection Agency, Salisbury, UK, but no longer listed in the American Type Culture Collection nt = not tested ART — source is the "Antimicrobials Resistance Team, CDC"
Inactivation of Bacterial Bioterrorism Agents in Water
19
------- |