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File: 950925_0pgv0014_oop.txt
MEMORANDUM FOR Defense Intelligence Agency, ATTN: DT-5C
Filename:0pgv0014.oop
AFMIC-S&TI (340d)
Subject: MEMORANDUM FOR Defense Intelligence Agency, ATTN: DT-5C
[ (b)(6) ], Washington, D.C. 20340-6053
SUBJECT: AFMIC Assessment Based on Laboratory Analysis of Iraqi
Serum Samples by U.S. Army Medical Research Institute for
Infectious Diseases (USAMRIID)
1. The 71 samples in question were obtained during
Operation Desert Storm. No medical history presently is available
on the individuals from whom the blood was collected. (See
enclosure 1).
2. Only a portion of the specimens were screened for
botulinum toxin neutralizing antibody. Methods used to screen the
group are contained in enclosure 1.
3. Results of analysis of the samples for immunization
against anthrax and botulinum toxin, possible Iraqi BW agents:
a. Anthrax--According to the attached USAMRIID report, during
initial serological screens, four sera of 69 were found to be
seropositive for Bacillus anthracis reactive antibody using the
enzyme immune assay (EIA). The EIA reactivity was confirmed by
western blot analysis. The sera contained reactivity against
protective antigen but not lethal factor by western blot analysis.
This suggests that the serum donors have been exposed to B.
anthracis antigens by natural infections or vaccinations. Having
proper histories (background, occupation, etc.) would help to
assess possible sources of natural exposure).
AFMIC cannot assess the immunization status of all Iraqi
soldiers against anthrax based on the information obtained from
exploitation of these samples alone. Because we do not know the
character of the Iraqi vaccine and because the kinetics of the
decay of antibodies post-vaccination for that vaccine is not
known, it is impossible to say unequivocally whether the antibody
titer is a result of active disease or immunization. However, it
can be stated that 1) not all Iraqi military are immunized against
biological warfare agents and 2) certain sectors of the Iraqi
military may receive immunization against anthrax.
b. Botulinum toxin-- A total of 7 samples were tested for
anti-botulinum toxin activity. [ (b)(1) sec 1.3(a)(4) ]
None of the 7 samples tested were found to contain botulinum
toxin neutralizing antibody at or above the 0.04 IU level.
None of the donor individuals whose sera were tested appear
to have been immunized against botulinum toxin. Since the
populatlon sample was small, no judgement can be made as to
whether the Iraqis are immunizing any of their troops against
botulinum toxin.
4. [ (b)(1) sec 1.3(a)(4) ]
5. Positive reactions obtained from sera testing for
Coxiella burnetti, Vaccinia virus, Sandfly fever virus, West Nile
virus, Rift Valley fever (RVF) virus, and Sindbis virus conform to
plausible expectations resulting from natural exposure to endemic
disease agents, routine vaccinations, or non-specific cross
reactions. The results from testing for these endemic diseases
has significantly increased the confidence level of AFMIC's
infectious desease threat assessment.
6. (U) AFMIC POC [ (b)(6) ], [ (b)(2) ]
[ (b)(6) ]
[ (b)(2) ]
United States Army Medical Institute for Infectious Diseases,
Special Pathogens Section (SPS), Department of Epidemiology,
Disease Assessment Division
SPECIMEN NUMBERS: [ (b)(2) ]
SPECIMEN DESCRIPTION. USAMRIID received, on 31 January 1991, 71
samples [ (b)(2) ]
collected during Operation Desert Storm.
ANALYSIS PERFORMED. [ (b)(1) sec 1.3(a)(4) ]
only a portion of the specimens were screened for botulinum toxin
neutralizing antibody. Since the chance of finding a specimen
seronegative for botulinum toxin serotype A neutralizing antibody
but seropositive for botulinum toxin serotype B, E or F antibody
is considered to be unlikely, the following procedure was used to
screen the 31 January survey group. First, a random 10% sample of
the sera was tested for neutralizing antibody against serotype A
botulinum toxin at a level of 0.04 IU. This level of antibody is
easily attained in man through immunization with a reasonably good
toxoid. Second, any sample found to be seropositive for serotype A
botulinum toxin neutralizing antibody was rescreened for serotype
B, E, and F reactivity at the 0.04 IU level. Third, any specimen
not in the 10% sample and found to be seropositive for B.
anthracis reacting antibody was tested for botulinum toxin
neutralizing antibody following the protocal described for the 10%
sample.
During the initial serological screens, four sera, [ (b)(2) ]
were found to be seropositive for B. anthracis reactive antibody
using the enzyme immune assay (EIA); the EIA reactivity was
confirmed by western blot analysis. Three sera, [ (b)(2) ]
were screened for botulinum toxin serotype A activity at a level
of 0.04 IU based upon these findings. Because of the small amount
of serum, the fourth sample [ (b)(2) ] was
depleted during the first series of test and could not be screened
for anti-botulinum toxin activity.
RESULTS. Serum samples [ (b)(2) ]
were found to be seronegative for botulirlum toxin serotype A
neutralizing activity and are considered not to contain botulinum
toxin neutralizing antibody at or above the 0.04 IU level.
[ (b)(6) ]
United States Army Medical Institute for Infectious Diseases,
Special Pathogens Section (SPS), Department of Epidemiology,
Disease Assessment Division
SPECIMEN NUMBERS: [ (b)(2) ]
SPECIMEN DESCRIPTION: USAMRIID received 71 serum samples on 31
January 1991. The samples were received in plastic freezer vials
wrapped in ziplock bags. The vials contained 0.3 to 3 mls and were
numbered [ (b)(1) sec 1.3(a)(4) ]
ANALYSIS PERFORMED: The samples were screened for evidence of
exposures or vaccinations against: botulinum toxin, B. anthracis,
F. tularensis, Y. Pestis, C. burnetii, vaccinia virus Crimean-
Congo hemorrhagic Fever virus, Rift Valley fever virus, sindbis
virus, West Nile virus, sandfly fever virus (Naples and Sicilian
strains), dengue virus, and Venezuelan equine encephalitis virus.
RESULTS SUMMARY: The results are presented in Table 1. None of
the serum specimens tested reacted with antigens from Cl.
botulinum, Y Pestis, F tularensis, CCHF virus, dengue virus type 2
or Venezuelan equine encephalitis virus. Varying numbers of sera
reacted with antigens from the remaining agents: 4/69 reacted with
B. anthracis, 7/69 reacted with C. burnetii, 8/69 reacted with
vaccinia virus, 1/69 reacted with Rift Valley fever virus, 18/69
reacted with West Nile virus 26/69 reacted with Sandfly virus
(Naples strain), and 55/69 reacted with Sandfly virus (Sicilian
strain).
RECOMMENDATIONS: [ (b)(1) sec 1.3(a)(4) ]
[ (b)(6) ]
RESULTS: Specimens [ (b)(2) ]
TABLE 1 TEST RESULTS SUMMARY ON 31 JANUARY SHIPMENT.
Serological Screen (a)
Agent Test (b) No.Pos/Total
(l)
B. anthracis EIA 4/69 (2)
Cl. botulinum toxin A NEUT 0/7 (3)
C. burnetii EIA 7/69 (4)
Y. Pestis EIA 0/69
F. tularensis MIA 0/69
Vaccinia virus IFA 8/69 (5)
CCHF virus EIA 0/69
RVF virus EIA 1/69
Sindbis virus EIA 3/69 (6)
West Nile virus EIA 18/69 (7)
Sandfly fever virus EIA 26/69 (8)
(Naples strain)
Sandfly fever virus EIA 55/69 (8)
(Sicilian strain)
Dengue virus Type 2 EIA 0/69
VEE virus EIA 0/69
(a) Numbers in parenthesis pertain to paragraphs in the Result
Interpretation section of the report.
(b) Abbreviations: EIA, enzyme immune assay; IFA,
immunofluorescent antibody assay; and MIA, microagglutination
test.
RESULTS INTERPRETATION: [ (b)(2) ]
1. Sample size. [ (b)(6) ][ (b)(2) ]
The vial numbers are listed in Table 2 along with corresponding
sample identification numbers from the CBW SAMPLE DOCUMENTATION
form. [ (b)(2) ]
[ (b)(1) sec 1.3(a)(4) ][ (b)(2) ]
2. Bacillus anthracis reactivity. Four specimens were positive for
B. anthracis antibody by EIA. The sera [ (b)(2) ]
contained reactivity against protective antigen but not lethal
factor by western blot analysis. The results suggest the serum
donors have been exposed to B. anthracis antigens by natural
infections or vaccinations. Consideration should be given to
conducting follow-up interviews of serum [ (b)(2) ]
donors; [ (b)(1) sec 1.3(a)(4) ]
history associated with the acquisition of B. anthracis reactive
antibody maybe important.
3. Botulinum toxin reactivity. [ (b)(2) ]
only a portion of the 69 specimens were screened for
botulinum toxin neutralizing antibody. Since the chance of finding
a specimen seronegative for botulinum toxin serotype A
neutralizing antibody but seropositive for botulinum toxin
serotype B, E or F antibody is considered to be unlikely, the
following procedure was followed to screen the 31 January survey
group. First, a random 10% sample of the nonprioritized sera was
tested for neutralizing antibody against serotype A botulinum
toxin at a level of 0.04 IU. This level of antibody is easily
attained in man through immunization with a reasonably good
toxoid. Second, any sample found to be seropositive for serotype A
botulinum toxin neutralizing antibody was rescreened for serotype
B, E, and F reactivity at the 0.04 IU level. Third, any specimen
not in the lO% sample and found to be seropositive for B.
anthracis reacting antibody was tested for botulinum toxin
neutralizing antibody using the protocol described for the lO%
sample.
Seven sera were tested for botulinum toxin serotype A antibody and
found to be seronegative. 3 of the 4 [ (b)(2) ]
sera found to be seropositive for B. anthracis reactive
antibody are presently being screened for botullinum toxin
serotype A activity. Because of the small amount of serum, the
fourth sample [ (b)(2) ] was depleted during
the first series of test and can not be screened for anti-
botulinum toxin activity.
4. Coxiella burnetii reactivity. Seven sera [ (b)(2) ]
were positive for C. burnetti antigen reactivity in the EIA
suggesting the serum donors had been infected in the past.
5. Vaccinia virus reactivity. Eight sera [ (b)(2) ]
reacted with vaccinia virus antigens in the IFA test. The
reactions were low titered; the titers were lower than that seen
in sera from U.S. troops serosurveyed two months after being
vaccinated against smallpox.
6. Sindbis Virus reactivity. Three sera [ (b)(2) ]
reacted with sindbis virus antigens in the EIA. The
seroreactivity was not unexpected since sindbis virus is endemic
in the Middle East and natural infections may be common in Iraq.
7. West Nile virus antigen reacivity Eighteen sera reacted with
West Nile virus (WNV) antigens in the EIA. The seroreactivity was
not unexpected since WNV is endemic in the Middle East and natural
infections may be common in Iraq. The WNV seroreactive specimens
are listed in Table 4.
8. Sandfly fever virus antigen reactivity. Twenty six sera reacted
with antigens from the Naples strain of sandfly fever virus and
fifty five sera reacted with antigens from the Sicilian strain of
sandfly fever virus. The seroreactivity was not unexpected since
these viruses are endemic in the Middle East and natural
infections may be common in Iraq. The results of the Sandfly fever
virus serology is prssented in Table 4.
TABLE 2. 31 JANUARY 1991 SHIPMENT SPECIMEN DOCUMENTATION.
SAMPLE NUMBER IDENTIFICATION NUMBER (1) VIAL NUMBER (2)
[ (b)(2) ]
TABLE 3 ISOENZYME LEVELS IN SPECIMENS WITH IDENTICAL NUMBERS
SAMPLE NUMBERS CPK DONORS
IDENTIFICATION NUMBER (l) VIAL N0.(2) LEVEL(3) IDENTITY NO.
[ (b)(2) ]
TABLE 4. SPECIMENS REACTING WITH WEST NILE VIRUS (WNV). SANDFLY
FEVER, VIRUS NAPLES (SFVN) AND SANDFLY FEVER VIRUS SICILIAN (SFVS)
ANTIGENS.
SAMPLE IDENTIFICATION NUMBER VIRUSES
WNV SFVN SFSV
[ (b)(2) ] - + +
[ (b)(2) ] - - +
[ (b)(2) ] - + +
[ (b)(2) ] - - +
[ (b)(2) ] - - +
[ (b)(2) ] - - +
[ (b)(2) ] + + +
[ (b)(2) ] - + +
[ (b)(2) ] - - +
[ (b)(2) ] - + +
[ (b)(2) ] + + +
[ (b)(2) ] + - +
[ (b)(2) ] + + +
[ (b)(2) ] - + +
[ (b)(2) ] - - +
[ (b)(2) ] + + +
[ (b)(2) ] - - -
[ (b)(2) ] - - +
[ (b)(2) ] - - +
[ (b)(2) ] - - +
[ (b)(2) ] - - +
[ (b)(2) ] + - +
[ (b)(2) ] - - +
[ (b)(2) ] + + +
[ (b)(2) ] - - +
[ (b)(2) ] - - +
[ (b)(2) ] + + +
[ (b)(2) ] + + +
[ (b)(2) ] - - +
[ (b)(2) ] + + +
[ (b)(2) ] + + +
[ (b)(2) ] - - +
[ (b)(2) ] + + +
[ (b)(2) ] + + +
[ (b)(2) ] + - +
[ (b)(2) ] - - +
[ (b)(2) ] - + +
[ (b)(2) ] + - +
[ (b)(2) ] - + +
[ (b)(2) ] - + +
[ (b)(2) ] - - +
[ (b)(2) ] - + -
[ (b)(2) ] - - +
[ (b)(2) ] - + -
(1). Symbols: -, negative reaction; +, positive reaction.
conti.
TABLE 4. SPECIMENS REACTING WITH WEST NILE VIRUS (WNV), SANDFLY
FEVER VIRUS NAPLES (SFVN), AND SANDFLY FEVER VIRUS SICILIAN (SFVS)
ANTIGENS.
SAMPLE IDENTIFICATION NUMBER VIRUSES
WNV SFVN SFVS
[ (b)(2) ] - + +
[ (b)(2) ] - - +
[ (b)(2) ] + - +
[ (b)(2) ] - - +
[ (b)(2) ] - - +
[ (b)(2) ] - - +
[ (b)(2) ] + - +
[ (b)(2) ] - - +
[ (b)(2) ] + - +
[ (b)(2) ] - + +
[ (b)(2) ] - + +
[ (b)(2) ] - - +
[ (b)(2) ] - + +
[ (b)(2) ] - + +
(1). Symbols: -, negative reaction; +, positive reaction
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