Spore Size Comparison Between Several Bacillus Species · Aspect Ratio (L/W) of Bacillus spores B...
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Spore Size Comparison Between Several Spore Size Comparison Between Several Bacillus Species Bacillus Species Ruben O. Zandomeni Ruben O. Zandomeni 1 1 , Joseph E. Fitzgibbon , Joseph E. Fitzgibbon 2 2 , Monica , Monica Carrera Carrera 1 1 , Edward Stuebing , Edward Stuebing 2 2 , James E. Rogers , James E. Rogers 2 2 , and , and Jose Jose - - Luis Sagripanti Luis Sagripanti 2 2 1 1 GeoCenters, Inc. and GeoCenters, Inc. and 2 2 RDECOM, Edgewood Chemical RDECOM, Edgewood Chemical and Biological Center, US Army, Aberdeen Proving and Biological Center, US Army, Aberdeen Proving Ground, MD 21010 Ground, MD 21010
Transcript of Spore Size Comparison Between Several Bacillus Species · Aspect Ratio (L/W) of Bacillus spores B...
Spore Size Comparison Between Several Spore Size Comparison Between Several Bacillus SpeciesBacillus Species
Ruben O. ZandomeniRuben O. Zandomeni11, Joseph E. Fitzgibbon, Joseph E. Fitzgibbon22, Monica , Monica CarreraCarrera11, Edward Stuebing, Edward Stuebing22, James E. Rogers, James E. Rogers22, and , and
JoseJose--Luis SagripantiLuis Sagripanti22
11GeoCenters, Inc. and GeoCenters, Inc. and 22RDECOM, Edgewood Chemical RDECOM, Edgewood Chemical and Biological Center, US Army, Aberdeen Proving and Biological Center, US Army, Aberdeen Proving
Ground, MD 21010Ground, MD 21010
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4. TITLE AND SUBTITLE Spore Size Comparison Between Several Bacillus Species
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Standard Form 298 (Rev. 8-98) Prescribed by ANSI Std Z39-18
•• GOALS GOALS -- Better understand properties of simulants Better understand properties of simulants vs. agents. Provide data to help choose appropriate vs. agents. Provide data to help choose appropriate simulants and surrogates for specific tasks.simulants and surrogates for specific tasks.
•• METHOD METHOD -- Compare properties of simulants to those Compare properties of simulants to those of the agents.of the agents.
IntroductionIntroduction
•• FACILITY FACILITY Microbiology Microbiology TeamTeamBSL3 BSL3 LaboratoryLaboratory
Properties (NonProperties (Non--Medical)Medical)
•• Spore Size Spore Size –– length, width, aspect ratiolength, width, aspect ratio
•• Spore DensitySpore Density
•• Effect of decon agentsEffect of decon agents
•• Fluorescence spectraFluorescence spectra
•• Effect of UV radiationEffect of UV radiation
Spore SizeSpore Size•• Important to design and development of samplers Important to design and development of samplers and detectors and detectors
•• Published spore sizes Published spore sizes -- spores produced under spores produced under different conditions for each species without different conditions for each species without extensive comparisons, size distributions or ranges.extensive comparisons, size distributions or ranges.
•• Systematic comparison of the size of Systematic comparison of the size of B.B.anthracisanthracisspores to size of other Bacillus spores spores to size of other Bacillus spores (simulants/surrogates) (simulants/surrogates) -- all spores produced under all spores produced under the same conditions.the same conditions.
MethodsMethods•• Prepared spores of Prepared spores of B.B.anthracisanthracis and other and other Bacillus species, including some common Bacillus species, including some common simulants/surrogates.simulants/surrogates.
•• Spores were fixed, negatively stained and Spores were fixed, negatively stained and imaged by TEM.imaged by TEM.
•• Measurements of 100 spores (length, width, Measurements of 100 spores (length, width, aspect ratio).aspect ratio).
•• Produced distributions.Produced distributions.
B. B. globigiiglobigii SB512SB512 (B. (B. atrophaeusatrophaeus))
•• Length Length
Mean 1.22Mean 1.22±±0.12 0.12 µµmm
Range 0.91Range 0.91--1.90 1.90 µµmm
•• Width Width
Mean 0.65Mean 0.65±±0.05 0.05 µµmm
Range 0.56Range 0.56--0.80 0.80 µµmm
B. globigii Size Disribution
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900 1000110012001300140015001600170018001900More
Length (nm)
Fre
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ency
.00%
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60.00%
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100.00%
120.00%
Frequency
Cumulative %
B. globigii Size Distribution
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Width (nm)
Freq
uenc
y
.00%
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100.00%
120.00%
Frequency
Cumulative %
B. B. subtilissubtilis 10311031
•• Length Length
Mean 1.07Mean 1.07±±0.09 0.09 µµmm
Range 0.87Range 0.87--1.41 1.41 µµmm
•• Width Width
Mean 0.48Mean 0.48±±0.03 0.03 µµmm
Range 0.41Range 0.41--0.58 0.58 µµmm
B. subtilis Size Distribution
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Length (nm)
Freq
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120.00%
FrequencyCumulative %
B. subtilis Size Distribution
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400 425 450 475 500 525 550 575 600 More
Width (nm)
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20.00%
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120.00%Frequency
Cumulative %
B. cereus B. cereus ATCC 10702ATCC 10702
•• Length Length Mean 1.55Mean 1.55±±0.16 0.16 µµmmRange 1.21Range 1.21--2.00 2.00 µµmm
•• Width Width Mean 0.90Mean 0.90±±0.07 0.07 µµmmRange 0.76Range 0.76--1.14 1.14 µµmm
B. cereus
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Length (nm)
Fre
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.00%
20.00%
40.00%
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80.00%
100.00%
120.00%FrequencyCumulative %
B. cereus
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Width (nm)
Fre
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.00%
20.00%
40.00%
60.00%
80.00%
100.00%
120.00%FrequencyCumulative %
B. B. megateriummegaterium WWWW--1515--49004900
•• Length Length
Mean 1.60Mean 1.60±±0.16 0.16 µµmm
Range 1.28Range 1.28--2.04 2.04 µµmm
•• Width Width
Mean 0.84Mean 0.84±±0.07 0.07 µµmm
Range 0.70Range 0.70--1.05 1.05 µµmm
B. megaterium
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Freq
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Frequency
Cumulative %
B. megaterium
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Frequency
Cumulative %
B. B. mycoidesmycoides ATCC 10206ATCC 10206
•• Length Length
Mean 1.85Mean 1.85±±0.21 0.21 µµmm
Range 1.33Range 1.33--2.44 2.44 µµmm•• Width Width
Mean 0.91Mean 0.91±±0.07 0.07 µµmm
Range 0.65Range 0.65--1.04 1.04 µµmm
B. mycoides
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Frequency
Cumulative %
B. mycoides
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Fre
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120.00%
FrequencyCumulative %
B. B. anthracisanthracis ((SterneSterne))
•• Length Length
Mean 1.53Mean 1.53±±0.15 0.15 µµmm
Range 1.19Range 1.19--1.92 1.92 µµmm
•• Width Width
Mean 0.88Mean 0.88±±0.06 0.06 µµmm
Range 0.71Range 0.71--1.09 1.09 µµmm
B. anthracis Sterne
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Length (nm)
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.00%
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Frequency
Cumulative %
B. anthracis Sterne
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Fre
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ency
.00%
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60.00%
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120.00%
Frequency
Cumulative %
B. B. anthracisanthracis ((∆∆--SterneSterne))
•• Length Length
Mean 1.56Mean 1.56±±0.16 0.16 µµmm
Range 1.23Range 1.23--2.05 2.05 µµmm
•• Width Width
Mean 0.82Mean 0.82±±0.06 0.06 µµmm
Range 0.68Range 0.68--0.98 0.98 µµmm
B. anthracis ∆-Sterne
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Fre
qu
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.00%
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Frequency
Cumulative %
B. anthracis ∆-Sterne
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Frequency
Cumulative %
B. B. anthracisanthracis (Albia)(Albia)
•• Length Length
Mean 1.43Mean 1.43±±0.14 0.14 µµmm
Range 1.22Range 1.22--1.79 1.79 µµmm
•• Width Width
Mean 0.87Mean 0.87±±0.07 0.07 µµmm
Range 0.71Range 0.71--1.03 1.03 µµmm
B. anthracis Albia
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2
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Fre
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.00%
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120.00%Frequency
Cumulative %
B. anthracis Albia
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.00%
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120.00%
Frequency
Cumulative %
B. B. anthracisanthracis (NCTC 1087)(NCTC 1087)
•• Length Length
Mean 1.68Mean 1.68±±0.19 0.19 µµmm
Range 1.23Range 1.23--2.08 2.08 µµmm
•• Width Width
Mean 0.89Mean 0.89±±0.06 0.06 µµmm
Range 0.76Range 0.76--1.09 1.09 µµmm
B. anthracis 1087
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Frequency
Cumulative %
B. anthracis 1087
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FrequencyCumulative %
B. B. anthracisanthracis (NCTC 1928)(NCTC 1928)
B. B. anthracisanthracis (LA1)(LA1)
Aspect Ratio (L/W) of Bacillus sporesAspect Ratio (L/W) of Bacillus sporesB anthracis Sterne
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1000 1200 1400 1600 1800 2000
Length
Wid
th
B globigii
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Length
Wid
th
B. subtilis
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Length
Wid
th
B. aB. a. NCTC 1087. NCTC 1087MeanMean 1.91 +/1.91 +/-- 0.230.23Range Range 1.371.37--2.452.45
B. subtilisB. subtilisMeanMean 2.23 +/2.23 +/-- 0.250.25Range Range 1.631.63--2.952.95
B. globigiiB. globigiiMeanMean 1.85 +/1.85 +/-- 0.190.19Range Range 1.431.43--2.392.39
B. aB. a. . SterneSterneMeanMean 1.75 +/1.75 +/-- 0.200.20Range Range 1.331.33--2.202.20
B. anthracis 1087
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1000 1200 1400 1600 1800 2000 2200
LengthW
idth
Comparison of Comparison of B.B. anthracisanthracis with other with other BacillusBacillus speciesspecies
Length (µm) Width (µm)
B. anthracis Albia 1.43 +/- 0.14 0.87 +/- 0.07 B. anthracis 1087 1.68 +/- 0.19 0.89 +/- 0.06B. anthracis Sterne 1.53 +/- 0.15 0.88 +/- 0.06B. anthracis D-Sterne 1.56 +/- 0.16 0.82 +/- 0.06
B. globigii 1.22 +/- 0.12 0.65 +/- 0.05
B. subtilis 1.07 +/- 0.09 0.48 +/- 0.03
B. cereus 1.55 +/- 0.16 0.90 +/- 0.07
B. megaterium 1.60 +/- 0.16 0.84 +/- 0.07
B. mycoides 1.85 +/- 0.21 0.91 +/- 0.07
B.a.B.a. 1087 vs. B. g. (Length)1087 vs. B. g. (Length)
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Length (nm)
1087
BG
B.a.B.a. 1087 vs. B. g. (Width)1087 vs. B. g. (Width)
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10152025303540
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B. a. 1087
BG
B.a.B.a. Albia vs. B. g. (Length)Albia vs. B. g. (Length)
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Fre
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Length (nm)
B.a. AlbiaBG
B.a. Albia vs. B. g. (Width)B.a. Albia vs. B. g. (Width)
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Mor
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10152025
3035
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Fre
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Width (nm)
B.a. Albia
BG
B.a.B.a. 1087 vs. 1087 vs. B. subtilisB. subtilis(Length)(Length)
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Mor
e
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Length (nm)
B. a. 1087
B. sub.
B.a.B.a. 1087 vs. 1087 vs. B. subtilisB. subtilis (Width)(Width)
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0
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Fre
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Width (nm)
1087
B. sub.
B.a.B.a. 1087 vs. 1087 vs. B. cereusB. cereus (Length)(Length)
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Mor
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req
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Length (nm)
B. a. 1087
B. cer.
B.a.B.a. 1087 vs. 1087 vs. B. cereusB. cereus (Width)(Width)
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750
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950
1050
1150
05
101520253035
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Fre
qu
ency
Width (nm)
1087
B. cer.
B.B. globigiiglobigii SB512SB512 (B. (B. atrophaeusatrophaeus))
•• Length Length
Mean 1.27Mean 1.27±±0.10 0.10 µµmm
Range 1.04Range 1.04--1.61 1.61 µµmm
•• Width Width
Mean 0.67Mean 0.67±±0.07 0.07 µµmm
Range 0.50Range 0.50--0.94 0.94 µµmm
B. globigii
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51015
2025
303540
4550
1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 More
Length
Fre
qu
ency
.00%
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Frequency
Cumulative %
B. globigii
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1020
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5060
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0.4 0.5 0.6 0.7 0.8 0.9 1 More
Width
Fre
qu
ency
.00%
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60.00%
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120.00%
FrequencyCumulative %
B.B. subtilissubtilis 10311031
•• Length Length
Mean 1.14Mean 1.14±±0.07 0.07 µµmm
Range 1.04Range 1.04--1.28 1.28 µµmm
•• Width Width
Mean 0.54Mean 0.54±±0.04 0.04 µµmm
Range 0.46Range 0.46--0.69 0.69 µµmm
B. subtilis
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0.9 1.05 1.1 1.15 1.2 1.25 1.3 More
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Fre
qu
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.00%
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120.00%Frequency
Cumulative %
B. subtilis
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0.4 0.45 0.5 0.55 0.6 0.65 0.7 More
Width
Fre
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ency
.00%
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60.00%
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100.00%
120.00%
Frequency
Cumulative %
B. B. anthracisanthracis ((∆∆--SterneSterne))
•• Length Length
Mean 1.48Mean 1.48±±0.07 0.07 µµmm
Range 1.37Range 1.37--1.71 1.71 µµmm
•• Width Width
Mean 0.89Mean 0.89±±0.05 0.05 µµmm
Range 0.75Range 0.75--1.00 1.00 µµmm
B. anthracis d-Sterne
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1.3 1.35 1.4 1.45 1.5 1.55 1.6 1.65 1.7 1.75More
Length
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ency
.00%
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FrequencyCumulative %
B. anthracis d-Sterne
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0.7 0.75 0.8 0.85 0.9 0.95 1 1.05 More
Width
Freq
uenc
y
.00%
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40.00%
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100.00%
120.00%
Frequency
Cumulative %
Comparison of Measurements Comparison of Measurements by TEM vs. OMby TEM vs. OM
Length WidthB. globigii TEM 1.22 +/- 0.12 0.65 +/- 0.05
OM 1.27 +/- 0.10 0.67 +/- 0.07
B. subtilis TEM 1.07 +/- 0.09 0.48 +/- 0.03OM 1.14 +/- 0.07 0.54 +/- 0.04
B. anthracis (d-Sterne) TEM 1.56 +/- 0.16 0.82 +/- 0.06OM 1.48 +/- 0.07 0.89 +/- 0.05
CONCLUSIONSCONCLUSIONS•• Systematic comparison of spore properties of several Systematic comparison of spore properties of several Bacillus species.Bacillus species.
•• Compared spore size properties of 6 species of Bacillus Compared spore size properties of 6 species of Bacillus by TEM.by TEM.
•• B. B. anthracisanthracis spores are significantly larger than spores spores are significantly larger than spores of the most common simulants, of the most common simulants, B. g.B. g. and and B. B. subtilissubtilis. .
•• B. B. anthracisanthracis spores are similar in size to spores are similar in size to B. cereusB. cereus and and B. B. megateriummegaterium..
•• Spores of pathogenic and nonSpores of pathogenic and non--pathogenic pathogenic B. B. anthracisanthracisare similar in size.are similar in size.
AcknowledgementsAcknowledgements
MICROBIOLOGY TEAMMICROBIOLOGY TEAMLeslie WilliamsLeslie WilliamsWarren GardnerWarren Gardner
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