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Karolinska Institutet and the Medical Research Council, Stockholm, Sweden

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Infection routes .

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Lines of defence .

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Sampling . . . . ... ...... .... . ..... .. , ................................ ,


Early warning . .

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Rapid identification. . ................................................


Protective equipment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..




Decontamination and disinfection. . . ............................ . ......


MEDICAL DEFENCE... . ..... ... ........................................


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Early diagnosis. .

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Chemoprophylaxis and treatment . . . .. ...... .... _ ..... ............. . . . . '


VETERINARY AND AGRICULTURAL DEFENCE. . ' ........................... '


"VATER, FOOD, AND DRUG PROTECTION...................................


RESTORATION MEASURES. . . ... . . _ ..... ............. .... .. ... . ... .... . . '


ADMINISTRATIVE MEASURES. . . .... ..... . ... ..... .. _ ... . . .. .... ...... .. '


Surveillance of communicable diseases.. . . ... . . ... ..... .... . ... . .. .. . . . . ,


Planning of biological warfare defence.. . ................................




The inhibitory forces which influence biological warfare developm<int... . ......


The case for disarmament. . ....................... ............. .. ... ..


Control and inspection problems. . ...................... .. . .. ...........



Many people, even some microbiologists, seem to be satisfied with gen

eral notions about biological warfare (BW) , and do not go beyond accepting
the fact that this imperceptible and insidious means of attack might affect
large groups of civilian as well as military personnel. As a result of a common
lack of knowledge of BW, until recently the subject has also largely been
avoided both in public discussions and by the authorities i n many countries.
When the question does arise, there is a tendency to treat it in a spirit of
apathy, since it is felt that the opportunities for any form of effective defence
are few or nonexistent. A short-lived reaction of this type has followed the
introduction of all "new" weapons since the crossbow, and it can be danger1

Prepared with the assistance of the Royal Swedish Academy of Engineering

Sciences as a working document for the Pugwash Study Group on Biological Warfare,
Marienbad, May 11-12, 1967.


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ous when exploited by an enemy. Since the psychological effects of BW

could be quite extensive, such an opponent might benefit from the fact that
unaccustomed methods of warfare tend to become the subject of sensation
alism and exaggeration and are often discussed in a very i maginative way.
One should not forget, however, that there is little reason to regard biological
agents as "ultimate weapons," and that there is a considerable time lag be
tween laboratory results and operational "hardware," even if the distance
may be smaller in the case of biological weapons than it is in the case of many
other types of armament.
The following summary is an attempt to view the possibilities of BW de
fence against the perspective of probable trends in the development of the
weapons on the one hand, and of international relations on the other. The
latter aspect is important because the political strain on an aggressor may be
j ust as important in BW protection as biological and physical defence mea
sures. Also, there is much to be said for the statement that the only defence
against chemical and biological weapons is early warning and international
agreements (1).
This review is an "outsider's" evaluation of scattered facts which are pre
sented in the hope that they will help the reader to reach a balanced view of
the military implications of modern microbiology. On the one hand, it must
be considered that there will be a risk of deterioration in the international
atmosphere if the daily press chooses to discuss the likelihood of BW in the
event of every major epidemic. On the other hand, a society which neglects
BW actually invites its use. Since a good portion of the medical and micro
biological professions in most countries tend to ignore the subject, the latter
risk is probably the largest, and this is a strong argument for a discussion of
the defence problems. Such a discussion might initiate measures aimed at
reducing the temptation to use BW, which an enemy may feel (2). This is
critical because once microbiological weapons gain acceptance, they could
initiate a very rapid escalation.
We are forced to record that as there are undeniable paranoid tendencies
in the relations between nations, and as the rate of change in society is often
more rapid than that of the learning process, communications tend to break
down. Also, the democratic voting mechanisms are clearly inadequate when
applied on the international level, and the gap between wealthy and poor
nations tends to grow. Consequently, the prospect for a lasting world peace
is not bright for the moment, and this puts special responsibilities on the
shoulders of the scientists. This has been spelled out in very clear terms by
P. Noel Baker (3):
Twenty years after the second 'war to end war', the generals as a class have shown
small belief that war and armaments can be abolished; they regard it, rather, as
their duty to explain the difficulties and dangers of disarmament; so far, very few
of them have made constructive proposals for ending the arms race.
The ministers have been willing.

year after year,

to accept resolutions in the General



Assembly of the United Nations declaring that disarmament is the most important

to give their delegates the

instructions which would enable them to negotiate a disarmament treaty. There
is no point in arguing which nations have been most to blame. The fact is that
ministers will only feel able to take grave decisions, which disarmament requires,
when there is an overwhelming body of public opinion throughout the world on
which they can rely to give them support.

objective of their foreign policy. But they have failed

Only the scientists can create this body of opinion. They can speak with an author
ity which the

ordinary citizen,

to which he
the generals or
endeavour to avert the

the nonscientist, cannot challenge, and

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is compelled to listen. Since they cannot hope for much help from
the ministers, they must act by

t hemselves, in a supreme

mortal dangers which confront ma nkind.

An indication that the dangers j ust mentioned might well include bio
logical warfare is given by the attitude of the superpowers.
Statements by leading Soviet military and political authorities are re
garded as indications that they are prepared to use, inter alia, biological wea
pons in a future war (4, 5) , but doubts about this interpretation have also
been expressed (6). It is reported that Defence Minister G. K. Zhukov said,
in February 1956, that a future war would li ke ly be fought with, inter alia,
nuclear, chemical, and biological (ABC) weapons. A Russian colonel added
in 1959 that "from results of comparative studies of the losses of life from
conventional weapons, war poisons, and atomic energy on one side and losses
from biological weapons on the other, it is believed today that a biological
war would have the greatest effect of all" (7). It has even been suggested
that BW might play a very important part in the Russian long-range offen
sive planning (7). Under all circumstances, Russia has set up a highly efficient
organisation for BW defence (8) and for the training of civilians in ABC pro
tection (9).
The United States was claimed to be behind the USSR both offensively
and defensively in 1962 ( 10) , but a five-year plan then existed which would
close the gap. As i ndicated by recent studies (11, 12), scattered i ndications,
like the use of 1236 monkeys in a determination of the median lethal dose of
i nhaled anthrax spores (13). and earlier reports on the American (14, 15) ,
British ( 1 6) , and Canadian ( 1 7) efforts in the field, the latter is attacked with
so much vigour that it would be most remarkable if no important develop
ments were to occur. After all, "the combining of military techniques and
science makes it easy to apply scientific principles to kill people-who are
not strong structures" ( 18) . Some of the results of purely basic research in
microbial genetics, molecular biology, and bioengineering also cast a sinister
light over Man's future as far as biological warfare is concerned. Add to this
the great advances made in microbiological laboratory personnel protection
(19-23) and the fact that equipment ranging from elaborate protection hoods
to fermentors suitable for the propagation of pathogen s ar e n o w co mmer
ci ally a va il a ble, a n d it s houl d become obvious that it is increasingly difficult
to neglect the BW d efence problems.



Throughout history, i nfectious diseases have been a serious military prob

lem, and in spite of antibiotics and the improved therapeutic means which

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became available during the second world war and in the Korean campaign,
infectious diseases caused several times more noneffectiveness among the
United States troops than battle injuries (24-26). Obviously, the deliberate
introduction of infectious diseases among troops would gain for the enemy
a military advantage, but BW has-with few exceptions (27)-been at
tempted in a crude way only, and never as a major weapons system (28) .
As late as the autumn of 1958, the American civil defence authorities re

garded the risk of biological and chemical attack as slight. Since then, how
ever, there has been a re-evaluation of the situation in that country, illus
trated i n a number of special i nvestigations ( 1 1 , 12, 29, 30) , which particu
larly considered the advances made in the handling of biological aerosols,
and put biological and chemical weapons in the same class as nuclear wea
pons. It was pointed out, for example, that military-political issues were be
coming more and more dependent upon civil defence pote nti al , which no
longer had the same secondary significance a s obtained during the era o f the
piloted bomber aircraft and conventional weapons. Also, experimental data
from human as well as animal studies provided background for such authori
tative statements as "the offensive use of biological agents is feasible" (26) ,
and "biological agents exist which can b e used strategically to cause casual
ties in an area the width of a con ti nen t " (31, 3 2) . Doubts concerning the
striking power of isolated infectious agents actually began to be dispelled dec
ades ago in tests on humans with, for example, Rickettsia tsutsugamushi (33)
via the skin , Plasmodium vivax via the blood stream (34), and Brucella
abortus (35) , i ntestinal bacteria (36, 37) , and poliomyelitis virus (38)
via the mouth. Of particular interest in this connection is the fact that the
aerosol dose of FranciseUa tularensis on humans could bc fixed at about 25
cells (39, 40). In the case of Q-fever, even a single inhaled parti cle (Coxiella
burnetii) might be sufficient to cause infection (41) , so that, in theory, a gram
of inoculated e m bryonic chicken tissue might hold no less than a billion hu
man infectious doses (42).
The possibility of dissemination over very large areas has also been dem
onstrated in field tests involving fluorescent particles (43) and bacterial
spores (44) . For instance, a cloud of 2/L zinc-cadmium sulphide particles
(450 pounds generated along a 15 6-mile stretch of coast) spread out over
about 34,000 square miles of land, where a minimum dose of 15 and a maxi
mum dose of 15 ,000 particles per minute were inhaled. I n the cited experi
ment with spores, 130 gallons of a suspension of Bacillus subtilis var. niger
was aerosolized from the deck of a ship running on a two-mile course about
two miles offshore. The dissemination line was at right angles to an onshore
wind, and meteorologically the situation was characterized by a certain tend
ency to vertical dilution. I n spite of this, the cloud could be followed for
about 23 miles, gi vi ng viable cell densities inside buildings corresponding to

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"infectious doses, " over some 100 square miles. Of course, a test of this sort
can be downgraded insofar as it concerned a spore, but guinea pigs have ac
tually been infected with vegetative bacteria which had travelled in an aero
sol nearly 15 miles (44, 45) .
A n ordinary aerosol generator, yielding a cloud in which only 5 per cent
of the particles were in the most infectious size range [1 to 5p. (46, 47) ], was
used in the spore experiment mentioned. If it had involved Bacillus anthracis,
of which, incidentally, 4130 spores is the LDoo for cynomolgus monkeys (13) ,
the size distribution would certainly have influenced the infectious dose,
since it is known from animal experiments that particles which are 12p. in
diameter give an infectious dose which is at least 17 times greater than that
which is required by a single cell aerosol [in this case which concerned guinea
pigs, about 23,000 cells (44)]. With regard to other types of disease agents
F. tularensis, C. burnetii, and Venezuelan equine encephalomyelitis) , there
may be a difference of many ten thousands in the requisite infectious dose
if one compares aerosols with 12 and 1p. diameters, respectively (44). The
latter types of particles are chiefly generated from dilute suspensions sub
jected to great force in fine nozzles and are consequently rare in nature (48,
From the foregoing it should be obvious that the offensive use of infec
tious clouds is intimately linked with the technology of generating fine-part
icle aerosols in which the virulence is maintained. If this technology is ade
quate, even the limited quantities of material which could be handled covert
ly might give disastrous results when disseminated over large areas.
Obviously, the size of aerosol generators which would be used in limited
attacks (on parliament buildings, military staff headquarters, etc.) would be
so small that they could easily be concealed by a saboteur. He could also
arrange to leave the scene in ample time before cases start to appear; in the
case of an attack with F. tularensis it would, for instance, take some two to
five days before the disease sym ptoms (fever, headache, malaise, sore throat,
muscular ache, and chest pains) would begin to make themselves felt (40).
The conventional image of biological warfare, the covert " man with the
suitcase"-poisoning water supplies and ventilation systems-may seem to
have been discarded (11), but this attitude might well prove to be premature,
at least if one considers specific situations; for instance, a sanitary break
down caused by nuclear attack or mobilization, when the psychological re
percussions of a covert BW attack might be very severe. It should not be
forgotten that as early as 1917 an agent succeeded in infecting 4500 donkeys
with Malleomyces mallei (27), and that a number of tendencies in a modern
society pave the way for sabotage acts: 1. extensive and rapid communica
tions, increasing the "coverage"; 2. urbanization; concentrating people an d
pr odu ction units in small areas ; 3. increasing the size of slaughter houses,
dairies, food processing industries and waterworks; 4. more efficient agricul
ture, i.e., extensive use of monocultures, large herds, and centralized fodder
manufacture; 5. central ventilation systems in command centres, subways,

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cinemas, theatres, restaurants, etc.; 6. increased dependence on key per

sonnel: radar and missile operators and the people in power, communica
tions, and transport centres. Such i ndividuals are frequently confined to
environments (shelters, submarines, and other navy units) in which conven
tional sabotage acts might be more difficult to carry out than the i ntroduc
tion of a biological agent with an incubation time giving a saboteur many
days lead time.
Also, in the case of attacks via food or water, the amounts of microor
ganisms or toxins that would be required to give substantial effects are small.
This is particularly true in cases in which multiplication can occur: typhoid
in-the-dairy type of attack, although, even where the agent does not i ncrease
in quantity, only relatively small amounts would be used. Drinking 100 ml
from a reservoir holding 5 million liters of water would involve a serious in
fection or an intoxication risk if half a kilo of Salmonella culture, less than
5 kg of a partiy purified botulinum toxin, or 7 kg of staphylococcal entero
toxin were to have been introduced (27) . This should be compared to the
" normal" poisons: five tons of fluorosodium acetate or ten tons of potas
sium cyanide would be required to give the same effect.
I t is just such effects, calculated per weight unit, plus the moderate pro
duction cost and the opportunities for anonymous use, which has given the
microbiological weapons the label "the weapons of the future. " Another con
sideration has centered on the "humanitarian" aspect of the incapacitating
varieties of biological weapons (10, 50, 5 1).
Biological wcapons are normally considered to be best suited for employ
ment against large scale-targets, and, considering some inherent character
istics such as the incubation period, they seem to lend themselves to strate
gic rather than to tactical purposes (52). However, the terminology is as
floating as in nuclear warfare, where advances in military technology have
obviously shifted the Hiroshima type of bomb from the strategic to the tac
tical class of weapons (3) . The fact that BW seems to offer particular promise
for counter-insurgency operations, and in fighting limited wars has probably
played a role in the considerations which led to the present state of develop
ment ( 1 1 ) . The possibility for anonymous use-even in "peacetime"- natu
rally adds a dimension of difficulty to the defence problems. Particularly i n
areas in which the epidemiological surveillance is i neffective and t h e public
health, veterinary, and agricultural services are weak, a systematic covert
plan to increase the incidence and spread of normally occurring infectious
diseases might be very difficult to expose and neutralize.
In a sense, the microbiological agents which might be used for offensive
purposes represent a whole range of weapon systems rather than a single
type of weapon. Antipersonnel, antianimal, and anticrop agents all pose dif
ferent defence problems, and an open aerosol attack would really be one wea
pons system and a covert vector dissemination, another. The situation is
further complicated by the various means which an enemy might employ i n
order t o potentiate a n attack. Radiation would aggravate the results o f an

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aerosol attack on man or animals, carriers like crystal needles might help
viruses to penetrate the epidermis of plants (27) , elimination of chlorination
would permit or simplify an attack via water, and so on. I t is obviously
very difficult to discuss the BW defence problems in anything but a hand
book, so the following text has been limited to microbiological agents with an
emphasis on aerosol warfare, antipersonnel weapons, and the international
aspects of BW defence. With regard to the defence against toxins from other
than microbiological sources, defoliating agents, plant hormones, etc., the
reader is referred to the more general texts which also give many additional
details on antipersonnel weapons (27, 53-56).

Much of the research and development work in biological warfare is kept

secret, less on account of its importance than because it is easily misunder
stood and provokes "emotional distress and moral turbulence" ( 1 1 ) . Efforts
to dampen such reactions by emphasizing the i n capaci tating agents in a "war
without deaths" have, however, involved a publicity which may have helped
to gain acceptance of the comprehensive chemical and biological warfare pro
gram which the United States began about 1960 ( 1 1 , 5 1 ) . In spite of this, the
attitude of the general public seems to be strongly negative over the world,
and much concern has been expressed among American scientists (57) , for in
stance, with regard to some contract research being conducted at academic
institutions ( 1 1 ) . From the defence point of view, such attitudes constitute a
danger, because the reaction is frequently so emotional that a proper per
spective on the need for defence measures is probably difficult to establish.
A psychiatrist once claimed that the common reaction to BW is similar to
that of the medical student who enters the postmortem theatre for the first
time; he can learn no anatomy until he has become accustomed to death.
Such an adaptation process is important for the defence planner who must be
able to create imaginatively a proper picture of conceivable BW-situations.

However, at the same time he should beware of the dangers inherent in the
of strategies which tends to alter the concrete nature of certain mili
tary problems by turning them into abstractions (overkill, megadeath,
megacorps, etc.), thus disguising the real nature of the operations from those
who take part in them (3). In this connection, the scientist has a special re
sponsibility, namely, to provide reminders that, for instance, "to restore a
situation with nuclear fire" might mean adding to the military chaos, and
"counter-force strategy" would mean destruction of enemy cities and the
eradication of their population. Although when, in outlining the dangers of
strategic jargon, Sir Solly Zuckerman emphasized nuclear war, he might have
pointed out that biological warfare of the genocidal type would certainly
create a similar situation.
Planning for conceivable BW situations is much more difficult than pre
paring for other types of disaster stich as earthquakes or train accidents, for
which there are precedents. With regard to BW, superstition has a tendency



to appear, and we hope that the threat will disappear if we adopt the ostrich
method of sticking our heads in the sand. This steals from an educational
effort the time which should be spent not only to induce watchfulness among
military, ci vil defence, customs, and target-indus try personnel, but also to
provide knowledge and experience with "exotic" infections to doctors, veter
inarians, plant pathologists, and microbiologists. The planning for defence

against biological weapons must be done long before it is recognized that an

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attack has taken place, and training is certainly essential if panic is to be

avoided. A BW-attack would be an acid test of the discipline and defence will
of a population.

Infection routes.-"To those who prefer the pleasant ostrich in-the-sand

attitude, rather than the blending of sane scepticism with stark reality, it

must again be pointed out that Koch's postulates have been confirmed in
Man many times for many disease agents as the result of inhalation of labor
atory-prepared materials"


Actually, th e potential for deliberate massive

induction of respiratory-acquired infection has been the focus of attention

among defence planners for many years. The subject of air-borne infection is
now a vast one and several excellent reviews and conference proceedings pro
vide a detailed treatment of the subject (59-63).
Aerosol weapons have a great diffusion capacity and they consequently
possess a "search" capability which is considerable. As with normal infec
tions, th e response of the i n dividual , however, varies within wide limits, in
dicating that much can be gained by increasing resistance at the physiologi
cal level, specifi c ally or nonspecifically (64).
Another infection route frequently discussed in BW reviews is via vec
tors. This is logical since the outcome of many wars has been determined by
such diseases as malaria and typhus. In fact, charges that this type of war
fare has been used against China have appeared on several occasions

(27, 65,

66). Military biologists are also reported to have achieved a great deal in the

BW purposes (51), and it has, for in

stance, been stated that a plant in Manchuria, during the last world war,
reached a production capacity of 45 litres of plague-carrying fleas per four
months period, i.e., the equivalent of 135 million insects (65). Toward the
field of mass production of insects for

end of the war, there were even plans for producing four times this quantity.
No less that 2000 arthropod vectors have been stated to transmit more

than 100 diseases to man (67), and this hints at some of the defence prob
lems to be found in this particular BW area. An effect on the vector as well
as on the infectious agent might be sought for at the physiological defence
In one experiment claimed to have been carried out in a relatively mos
quito-free district in Florida (51), 200,000 mosquitoes were liberated, and it
was found that a large portion of the personnel at an airbase suffered several
stings in the space of a couple of days. If these insects had carried yellow

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fever, a considerable proportion of the base personnel would probably have

been infected (68) . Tests on volunteers have shown that malaria (69) and
dengue fever (70, 71) could also be spread in this way.
Lines of defence.-In an aerosol exposure, the first line of physiological de
fence is provided by the nose and nasopharynx (72), which serve as efficient
traps for particles larger than 10 p. and may even retain as much as 50 per
cent in the 1- to 5-p. range (73). In cases in which the local defence is not over
whelmed (d. tonsillar plague) , or where there is not a tropism for the respira
tory mucosa (d. influenza virus) , deposition in the upper respiratory tract
rapidly renders the microorganisms innocuous (74) . The selectivity of the
upper respiratory tract, and the dependence of the clinical picture on the
place of deposition, have been illustrated both i n volunteers exposed to bac
teria (40) and viruses (7S) . Experiments on rhesus monkeys exposed to Fran
cisella tularensis are also very illustrative (76) . Here, 18-p. particles caused
lymphadenitis in the nasopharynx drainage area followed by dissemination,
whereas a 1- to 8-p. particle aerosol caused bronchiolar involvement. The size
also had a profound influence on the LD50, which fell from 3000 at 22 J1. to
240 at 7 p., and 17 at 1 fJ.. The range was even greater for guinea pigs, which
are less prone to mouth breathing than monkeys and also have a greater fil
tering capacity in the upper respiratory tract. A similar dependence of res
piratory infectivity upon particle size has been demonstrated with Bacillus
anthracis spores, Coxiella burnetii, Brucella suis, and the virus of Venezuelan
equine encephalomyelitis (74) .
A second line of defence is provided by the mucociliary lining which
sweeps entrapped particles out of the tracheobronchial tree. In the finest bran

ches, the respiratory bronchioles, there is, however, no lining, and particles
deposited in this region [preferentially the 1- to 5-p. range (7 7-80)] persist for
a considerable time.
A third line of defence is the phagocystosis which occurs in the depth of
the lung (73, 81). If this defence is overcome or bypassed, either by the viru
lence factors of the microorganism or by other influences on the host [a pre
vious virus infection, hypoxia and acidosis, ethyl alcohol, or tobacco smoke
(8 1)], there may be either an initial local involvement as in tularemia (82)
and psittacosis (83) , or the microorganisms enter the vascular system directly
as in Q-fever (41 ) . Phagocystosis may, however, be a factor in penetration as
well as in protection. The former case is illustrated by the finding that cells
which loose their Vi antigen-and thus become more easily ingested-also
infect laboratory animals via the respiratory tract more easily (84).
The fourth line of defence-nonspecific immunity and specific antibodies
-can hardly be called a line because it operates throughout the organism of
the host, on the nasal mucosa (85) and deep in the tissues, even if bactericidal
powers in the blood are the most dramatic. This defence can often be
strengthened through immunization, a possibility which should always be
considered when there is sufficient time for a response.
Immunization may play a role in the defense against a large number of

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potential BW agents, irrespective of their route of entry (19) . However, there

is a limit to the number of vaccines available and to the quantities of antigen
which are tolerated by man, even if much can be gained by purification. Con
sequently, even if it were to become one of the most important elements in
the BW-defence (45 ) , a full immunization program will probably never be
come a general BW-"prophylactic." It seems more realistic to aim at the
limited goal of immunological orientation against a multiplicity of antigens
rather than at a full immunity against all of them (86) . Intelligence informa
tion about sudden additions to the immunological protection which an ex
pected enemy gives his own troops would, of course, influence a military
vaccination program.
There exist attenuated living strains suitable for use as vaccines against
many potential B W-agents (86) (F. tularensis, Pasteuralla pestis, Brucella
abortus bovis, B. anthracis, Rickettsia prowazekii, and the viruses of variola,
yellow fever, Venezuelan equine encephalomyelitis and poliomyelitis) , and
others, are being developed (West Nile virus and members of the Russian
spring summer virus group). However, such vaccines often pose problems of
stability upon storage, so efficient nonmultiplying antigens are also sought,
even if they occasionally also present stability problems, as in the case of the
vaccine against Japanese encephalitis (86) . The nonliving vaccines range
from the highly efficient toxoids (against tetanus, diphtheria, and botulism)
and certain viral and rickettsial preparations (against rabies, typhus,
adenovirus, influenza, and Rocky Mountain spotted fever) , over bacterial
vaccines (against typhoid, cholera, and plague) to relatively inefficient
immunizing agents (against paratyphoid and shigellosis) . In some cases, very
notable advances have been made in military laboratories which can, for in
stance, claim the development of a multivalent toxoid that gives simultane
ous protection against the five common botulinum toxins (10) , excluding only
some novel types (87) . The work on continuous cultivation and that on the
biochemical basis of pathogenicity in Brucella, performed at the Microbiolog
ical Research Establishment at Porton in England, have been characterized
as two of the most significant advances in microbiology made over the past
20 years (16) . In the case of anthrax, both the testing (88, 89) and the culti
vation methods (90) were greatly improved in military laboratories, and in
the case of tularemia (9 1 , 92) and Q-fever (93), military studies on human
volunteers proved essential for the vaccine evaluation. In the latter case, the
subjects were challenged by measured low doses of aerosolized C. burnetii, a
microorganism which normally reaches man via the respiratory tract. I m
munization proved to be very effective, and even a single dose of vaccine
given to nonvaccinated personnel soon after exposure was of some value, par
ticularly if immunity was permitted to develop while the disease was sup
pressed with tetracycline. The same observation has been made in the case of
inhalation anthrax in monkeys treated with penicillin. In itself, this antibiot
ic will only delay the appearance of symptoms (94).
As far as mass immunization techniques are concerned, the jet injector

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has great possibilities but in some cases such as tularemia, aerosol vaccina
tion seems to be an attractive field technique (92, 95-102) which would theo
retically be preferable where the expected challenge will be via the respira
tory route (103, 104). In fact, it may well develop into one of the most im
portant vaccination techniques of the future ( 1 05). However, systemic reac
tions may occur (92, 96, 106, 107); the dosage presents a problem (92) and
more knowledge regarding a conceivable dissociation of viability and viru
lence is required (58). Also, the interesting possibility of increasing the
response to inhaled toxoids (108-111) with the aid of adjuvants (112) needs
further investi gation. Particularly, the Russian studies of aerogenic immuni
zation of animals and man with dried living vaccines mainly against anthrax,
brucellosis, plague, and tularemia (107, 113, 114), and American studies on
mixed tularemia and Venezuelan equine en ceph alitis vaccines (115, 1 1 6) are
highly relevant in the context of this review.

Detection is a prerequisite for any defensive action, and it can be sub

divided into three elements: warning, sampling, and identification (117) . An
early warning will trigger the use of personnel protective equipment as well
as preparations for identification. A direct defence against an aerosol attack
by means of explosions or defence aerosols has also been discussed (27, 118) ,
but the efficiency of such measures is difficult to j udge.
In many cases, the population in the target area might be unaware of an
attack until the appearance of clinical cases, but this constitutes an intelli
gence warning rather than a basis for warning in the normal sense of the
word. However, some information (about symptoms, chemoprophylactic
treatment, etc.) would also be issued in this case in order to prepare the tar
get population.
Early warning.-A warning could be based on the appearance of delivery
systems (planes or m issiles) which do not behave as normal missions would be
expected to do. Another warning might be the appearance of nonindigenous
arthropods or large numbers of vectors at a time or in a place where they
would not be anticipated. However, visual observations would never consti
tute a reliable basis, because a water and food attack might be launched
anonymously far from the target area, and aerosols would probably be re
leased along a line at a distance from it. Obviously, there is a case for a warn
ing network that would issue commands dealing with masking, entering
shelters, water and food treatment, and the taking of meaningful samples.
In the case of a grid set up to detect aerosol attacks, the systems aspects are
complex since population densities, target importance, terrain parameters,
prevalent meteorological conditions, and varying types of dispersion (moving
or stationary point source(s), one or more primary clouds generated simul
taneously or in succession etc.) must all be considered (119). Warning devices
that are so small and simple that they could be issued to small groups of in
dividuals, belong to so distant a future that they need not be considered here.

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Actually, the problems involved in the warning against an aerosol attack,

for instance, are so formidable that the development might call for regional
or international co-operation between many countries ( 120) .
The desirability of stationing warning devices in "high risk" areas would
be an argument for the participation of some countries from the developing
parts of the world. As emphasized within the Pugwash Organization, inter
national cooperation will also help to reduce mistrust between nations (121,
One of the technical difficulties is the normal background material, for
instance, mold spores, and this changes with the time and place, making it
necessary to base a setting of the warning threshold on a thorough knowledge
of the local situation. As far as the United States is concerned, the concen
tration of viable aerobic bacteria is usually less than 1 per litre of outdoor
air, and the protein content is about 3 X lQ-Sgjlitre ( 1 1 7 ) . Automatic warn
ing devices would have to measure continously such minute quantities of pro
tein or nucleic acid or both, and in itself this involves a range of technical
problems (123). Moreover, the particle content in a narrow critical-size
range would have to be monitored.
At the present time it seems necessary to combine several principles of de
tection (124) and to depend on human integration of many factors, including
meteorological conditions, in order to issue an alarm. Many sophisticated
warning systems are conceivable [long-path light scattering ( 125) and infra
red absorption, dielectric particle counting, gas chromatography (126) , acri
dine-orange staining, enzymatic detection of cell wall D-alanine ( 127)], but,
considering the multitude of routes available for delivery and the great num
ber of agents which the enemy might choose, it is highly i mprobable that
any system will ever be able to prevent the occurrence of significant numbers
of clinical cases.
Sampling.-The method of sampling depends, of course, on the material
studied, and such established microbiological techniques as membrane fil
tration or swabbing, can be used for water, milk, foodstuffs, and so on. For
aerosols a large variety of samplers are also available ( 1 28-133) , some of
which could be triggered by an early warning device and set up to make a
size discrimination ( 134) . Among recent samplers is one that mimics the par
ticle size discrimination of the respiratory tract but stilI has a high sampling
rate and yields a high concentration of cells per unit of collecting fluid ( 135).
Another has a very high capacity, collecting 10 ma of air per minute into a
small amount of liquid ( 13 6) . The latter opens up a whole new field since one
might theoretically detect airborne agents in one-thousandths of the concen
tration detectable by conventional samplers.
Once it is recognized that a biological agent has been used, suitable speci
mens, together with a report giving all relevant information, must be rushed
to the laboratory. In order not to lose cultivation time during the transport,
a portion of the suspected material can be used to inoculate suitable media
immediately after sampling. They are then sent to the laboratory in temper-



ature-controlled boxes or stuck in the pockets of special waistcoats designed

to make use of the body temperature of the wearer.
Both the collection of samples, which should be done by specialized mo
torized patrols, and the transport arrangements, call for a definite plan of
action since only one or a few reference laboratories might be equipped to
carry out the special serology, animal inoculations, tissue and egg inocula
tions that are required (137). Good judgement must be shown by the field

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personnel in their selection of specimens, so that the central facility is not

overwhelmed with unrewarding material, i.e., specimens which are not prop
erly collected or are likely to contain the same agent. As in the United States,
one might also have to consider transporting a few selected patients to a cen
tral location, where the coordinated efforts of highly competent clinicians and
laboratory personnel might shorten the time in which a diagnosis can be es
Potentially infected insects should, of course, also be collected and trans
ported to a laboratory equipped for identification work (27, 67) . It should be
borne in mind that such agents could be disseminated both covertly and
overtly, and that not only could normal transmission be mimicked, but con
ceivably also a vector-agent combination might be used which is not nor
mally seen.

Rapid identijicati on . Iden tification is necessary in order to facilitate a

purposeful treatment and to estimate in good time the number of casualties
and their regional occurrence. This requires an understanding of the charac
ter of conceivable weapons if the laboratory efforts are to be properly organ
An agent suitable for military use would probably belong to the group of
microorganisms which tend to produce laboratory infections (138, 139) . This

includes F. tularensis (40, 91) , C. burnetii (41), R. prowazekii, and the causa
tive agents of psittacosis, Venezuelan equine encephalomyelitis (140-142) ,
and Russian spring summer encephalitis, t o name a few. However, the agent
would also have to be suitable for large-scale production, storage, and dissem
ination. Review articles (44, 143-147) actually list a large number of micro
organisms which the identification laboratory should be able to recognize
(d. al so table in section on chemoprophylaxis and treatment) . This list com
prises about a dozen bacteria [for instance, those causing anthrax (148, 149) ;
plague (150) ; tuberculosis (151) ; glanders, brucellosis, cholera, and dysen
tery], some fungi [causing coccidioidomycosis (152, 153) ; histoplasmosis

(154, 155)]. protozoa (malaria); and a group of viruses and rickettsia (146)
[for instance, those causing influenza (156-158); smallpox, yellow fever (159);
Rift Valley fever (159); Japanese B encephalomyelitis, foot-and-mouth dis
ease, and Rocky Mountain spotted fever (158, 160)], apart from those men
tioned earlier. In addition, there are a number of toxins, i.e., botulinum toxin,
which may be even more effective in the aerosol form than if the same dose
were taken orally (161, 162). However, some of the agents can be placed far
down the list of probable weapons and this simplifies the situation. There are

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several reasons for this. For example, it is unlikely that an aggressor would
wish to create a permanent memorial to his BW campaign by using such dis
figuring diseases as smallpox (5 1) which, in most cases, can also be dismissed
because of the extensive use of vaccination. A reason making an agent un
likely as a weapon is that effective prophylactic methods are available.
The identification laboratory must bear in mind the possibilities of char
acter manipUlation, but the development of microorganisms which would
produce entirely new diseases may not be as important as current speCUla
tion would indicate. " New, " i.e., previously unreported virus diseases, con
tinually appear (d. Chikungunya and O'Nyong-Nyong fever) , and they
might be j ust as suitable for BW purposes as any product of laboratory
manipUlation. However, virulence can occasionally be increased, character
istic identification features can be eliminated, and antibiotic resistance can
easily be induced (163, 164) , and it is therefore certainly foolish to down
grade the conceivable military applications of modern microbial genetics, in
particular, transduction with resistance transfer factors. The possibility of
infection by streptomycin-resistant F. tularensis ( 165) could be mentioned,
but it is likely that a wide spectrum of potential biological weapons already
exist in, or could be produced in, an antibiotic-resistant form.
Speed in the identification process is essential, since large groups of indi
viduals would have to be detained, and also because occasionally it might be
too late to institute treatment by the time a clinical diagnosis is made. In the
case of i nhalation anthrax, treatment should start when the symptoms are
vague and before the condition becomes alarming ( 166). Pneumonic plague
has a mortality close to 100 per cent if treatment is not instituted within 20
to 24 hours after exposure ( 167-169) , and early treatment is also important
in the case of tularemia (170) . Obviously, the speed of identification required
in such cases is a challenge to the microbiologist who would have, only under
exceptional conditions (recovery of defective munitions, expended spray de
vices, vector containers, etc.) more than a minute amount of material avail
able for study immediately after an attack. Actually, there is room for much
research on microcultivation techniques (171), improved growth media (172)
phage techniques, fluorescent tagging of antibodies and antigens, blocking
antibody determinations, testing of sera for viral antigens instead of anti
body, specific gamma globulin consumption tests, and microserological
techniques in general ( 173) , together with all of the supporting instrumenta
tion needed to improve accuracy and to simplify operation (174). Gas chro
matography (126) and nucleic acid hybridization methods (175, 176) might
also deserve attention, but most efforts seem to be concentrated on the fluo
rescein-labelled antibody technique ( 124) which can be automated ( 177) and
brought to a remarkable sensitivity. It has, for instance, been possible to
identify tularemia organisms on glass slides from a cascade impactor which
had operated for only two minutes at a concentration of 1 bacterium per 5
litres of air (178) . The same technique has also been successfully employed
for the identification of aerosolized Venezuelan equine encephalomyelitis vi-

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rus ( 1 79). This agent as well as the virus causing Rift Valley fever could also
be detected in tissue cultures within 24 hours of propagation ( 1 79) . I n addi
tion, the technique has proved to be superior to conventional staining meth
ods in the search for the Negri bodies of rabies ( 1 80) . In comparison to iso
tope- and ferritin-labelling of antibodies, the fluorescent antibody technique
seems to offer considerable promise, particularly for identification after pri
mary isolation of many potential biological weapons ( 1 79, 1 8 1 ) .
The existence of sophisticated methods should not exclude the parallel
use of conventional techniques ( 1 7 1 , 182, 183) such as the direct staining of
sputum which might disclose cases of pneumonic plague. In most cases, rel
atively large numbers of bacteria are required for microscopic examination to
be effective. Even in the case of viral diseases, the conventional methods will
probably constitute the backbone of laboratory diagnosis for a long time to
come ( 1 80, 184, 185) .
Protective equipment.-Protection against aerosols will, to large extent,
depend upon the availability of efficient masks or over-pressure ventilation
with filtered air. Military masks normally remove all particles (or 99.99 per
cent in the i-to 5-}.' range) . M any filter materials have been shown to remove
microorganisms very effectively ( 1 86), but the complete mask should be thor
oughly checked for leaks by biological simulants ( 1 8 7 ) . I n many countries
excellent masks are also available for the civilian population, and even in
fants can be protected in a special tent-like structure ( 1 1 7 ) . I f masks are not
available, a man's handkerchief folded as thick as possible and held tightly
over the mouth and nose might remove more than 90 per cent of the organ
isms that would otherwise be inhaled ( 1 88) . The common hospital type mask,
on the other hand, removes only 20 per cent ( 1 89) .
Special positive-pressure shelters can make masks unnecessary and pro
visional sealing of rooms may be helpful, but normal buildings offer very
little protection (45, 1 1 7) .
I n case of suspected vector attacks, different types of repellents, bed nets,
protective clothing, etc., are important ( 67 ) .
Decay.-All disseminated infectious agents are subject to a relatively
rapid loss of viability and virulence, and the rate of decay must be estimated
if the protection and decontamination measures are to be given proper scope.
At the moment of aerosolization, the microorganisms in a suspension are
subjected to considerable stresses i n the form of heat, pressure, and shearing
action, and their preparation in the form of a powder, which can be aeroso
lized, is attended by similar forces ( 190) . The viability of vegetative cells con
sequently drops rapidly, particularly if they are not properly protected ( 1 9 1 ,
192), and it decreases even further during the short period in which they
equilibrate with the humidity in the air. Quite small changes in relative hu
midity may, in fact, drastically affect the viability of air-borne bacteria ( 193,
194), and it has been suggested that the movement of bound water in and out
of the cell results in a collapse of the protein structure ( 195, 196) . However,
oxygen also plays a significant role ( 197) and metabolic processes probably

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65 4


take part. Some organisms are very sensitive to extremely high or low humid
ities (76, 198) , but spores and some rickettsia seem to be relatively resistant,
as indicated by outbreaks of Q-fever and some plant diseases far removed
from the points where the particles became air-borne (199, 200) . However,
even within one group of such microorganisms as the rickettsia, the aerosol
stability varies from relatively high for Q-fever to the relatively low for R.
rickettsii and R. prowazekii suspensions (201). Some viruses, like Coxsackie
A-2 1, might decay at a rate of 25 to 50 per cent per minute depending on the
particle size (202) , but with the proper suspending fluid, survival can occas
ionally be increased (203) .
During their journey in a cloud, microorganisms are also exposed to oxy
gen and irradiation. Since most pathogens suffer a rapid reduction of viability
when exposed to light (204, 205), dissemination would probably be limited to
the hours of darkness and preferably to cool weather when decay is slower
(206). Photoprotection is a possibility which might be used in the case of
smaller particles.
Beside the "biological decay," which is attributed to such factors as those
mentioned, an aerosol cloud is also subject to "physical decay, " i.e., a reduc
tion in the effective particle concentration resulting from dilution, settling
out, rainfall, and impaction upon surfaces.

Atmospheric dilution depends upon the meteorological conditions and

the terrain. If an aerosol is generated along a line perpendicular to the wind
direction, lateral diffusion is significant only at the ends of the cylindrical
cloud, and diffusion along the wind direction does not alter the total dose pre
sented (207) . Vertical mixing on the other hand may cause a rapid drop in the
particle concentration, and this is why an aerosol attack is most likely to oc
cur on clear nights when the earth's surface is cooled by radiation and there
is a "lid" of warm air to limit the upward movement of the aerosol (208, 209).
Dissemination could be limited by the topography of the terrain, but very
large areas might be involved in so-called polar outbreaks where a 3000- to
6000-feet thick layer of cold air could serve as a carrier for an aerosol moving
over hundreds of miles at speeds of 20 to 25 miles per hour (3 1, 32) .
Loss of particles as a consequence of settling out or precipitation is rela
tively small in the size -range likely to be used in aerosol warfare. It has, for
instance, been estimated that an inch of rainfall would remove only 10 per
cent of the particles i n the 3-J.L range (209) . Also, the losses caused by impac
tion are probably small, but available test data do not permit detailed pre
The effects of BW agents, calculated on a weight basis, are such that an
attacker in many cases can compensate for the decay even if only standard
protection (76, 2 10-2 14) is provided and due consideration given to the phys
iological state of the microorganisms at the time of aerosolization (215).
Using a combination of sugars and metal-binders it has been possible to make
aerosolized Serratia marcescens nearly as stable as spores of B. subtilis var.
niger ( 19 1). If F. tularensis were aerosolized the infectious dose would be

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about 25 cells ( 2 1 6) , and it might perhaps be assumed that the infecting ca

pacity on humans falls off at the same rate as it does on guinea pigs. Initially,
the infectious dose is 10 to 20 cells, but it increases to about 150 to 200 when
the aerosol is five and a half hours old (76). A relatively small amount of
material would obviously be required to establish this dose over hundreds of
square miles.
Theoretical calculations clearly indicate the possibilities for large-scale
coverage. A midnight dispersion of 5 litres per kilometer of a suspension hold
ing 1010 particles/ml at an altitude of 100 meters along a 50 kilometer line
would-given a reasonable generator efficiency, certain meteorological condi
tions, and a wind speed of 20 kilometers per hour-set up a cylindrical cloud
which would pass a downwind point in less than a minute (207) . A person
breathing at a rate of 10 litres per minute would be exposed to about 150,000
particles. If this happened at 2 o'clock in the morning and if the relative hu
midity had been appropriate for the selected agent (decay : 2 per cent per
minute) , an aerosol with an infectious dose of 150 viable particles might be
enough to cause disease. An individual exposed at 6 o'clock would, on the
other hand, only contract the disease if the agent used had an i nfectious dose
of 1 . 5 particles. At this time the coverage would be 6000 square kilometers.
Decontamination and disinfection.-The gaseous sterilants ethylene oxide
(217) and i3-propiolactone (218) in particular, have attracted attention as
BW decontaminants (2 19) . The former is difficult to contain and the con
taminated material must be exposed in cabinets or plastic bags (220). This
gas easily penetrates porous materials, whereas i3-propiolactone primarily
sterilizes exposed surfaces, making it specially useful for the decontamination
of large interior spaces (22 1 ) . Decontamination of extensive areas is hardly
practical. Rather, natural decay, aS5isted by sunlight, temperature, and air
movement must be relied upon (222-224) .
Against supposedly infected arthropods, quickly acting insecticides would,
of course, be used.
In an attack on a city, a large depot or a major industrial or communica
tion center, an enemy would probably make no distinction between military
and civilian populations, and consequently the development of an adequate
medical defence is as important to the civil defence sector as to the armed
forces (26) . Clearly, preparations for disaster medical care are required (2,
225) , involving stand-by facilities such as "Packaged Disaster Hospitals"
available in the United States (226) . Some 2000 such hospitals are stra
tegically located across the continent, each being able to care for about 200
patients for 30 days.
Early diagnosis.-As indicated in the section on sampling, close coopera
tion between clinician and laboratory might reduce the time required for a
diagnosis to be made. Even if the initial phase of many infectious diseases is
s o uncharacteristic that the first clue can be detected only in the laboratory



the roentgenologists (227) might give useful hints in aerosol attacks involv
ing agents such as F. tularensis ( 1 65 , 2 28) , P. pestis (229, 230) , B. anthracis
( 1 66) and C. burnetii (42, 231).
Biological weapons might well present a clinical picture which is n ew and
unexpected since exotic or mixed diseases may be induced, or because the
agents may circumvent the normal routes of entry. I t

has been pointed out

that certain mixed infections could present a clinical picture that might de
lay the institution of proper therapy.

A nonresponsive viral disease might

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go into a recovery phase simultaneously with the appearance of symptoms of

a high mortality if left untreated

(232) . Respiratory infections with the bacteria causing anthrax, plague, and
tularemia would also produce a different clinical picture from that seen nor
mally. Medical experience might thus be a drawback, since it would tend to
exclude a particular diagnosis. The education and training of medical per
sonnel who could become involved in -BW defence should take this risk into
In an area where A edes aegypti is absent, one would probably not be as
a bacterial disease which would lead to

alert in diagnosing the first few human cases after a small-scale aerosol at
tack with yellow fever virus, as one would be in a region where the presence

of the vector induces a watchful attitude. In the latter case, one would look
for suspicious cases and might even have a routine for histological examina
tion of the livers of people dying within 10 days after the onset of any febrile
disease (233). J u dgi ng from American studies on Macaw mulatta, needle
biopsy in combination with fluorescent antibodies might even give a diag
nosis of yellow fever during the first few days of illness. The bleeding ten
dency associated with this disease (234) would, however, not permit a wide
spread use on humans, but the approach might be very useful for obtaining a
pointer both in this and in other infectious diseases (179) .
Other potential aerosol weapons which might offer difficulties similar to
those encountered with yellow fever, are the normally arthropod-borne
agents of Venezuelan or other equine encephalomyelitis, Rift Valley fever, and
Rocky Mountain spotted fever. This type of weapon is particularly i mpor
tant in

BW defence since one must assume that an attacker would regard the

lack of secondary cases as a considerable advantage. As compared with those

sols would cause diseases that are more easily recognized : psittacosis, Q
fever, smallpox, histoplasmosis, and coccidioidomycosis (74) .
Chemoprophylaxis and treatment.-It would be impractical, wasteful, and
dangerous to use the "shotgun" approach of placing everyone on prophylac
tic antibiotics in anticipation of an attack (137) . However, the situation
might change somewhat with the appearance of long-acting antibiotics (232) ,
and there should always be a stock of broad spectrum antibiotics available
for use when an attack h as occurred and the agent has been identified. Con

viruses and rickettsia, the agents which are normally transmitted as

sidering that strains made resistant to a conventional therapeutic approach

might well have been used, the sensitivity pattern must always be deter-

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mined. When the supply of antibiotics and chemotherapeutics is scarce,

treatment should normally be delayed until that pattern is available. The
timing of the therapy must then be carefully considered since too early a
treatment might only suppress the disease [d. Q-fever (42)] ; and too brief an
administration might not effect a cure [d. anthrax (94) , and tularemia (235) ].
Chloramphenicol and tetracycline are effective in the treatment of scrub ty
phus, but they will not prevent the disease if administered in standard daily
doses during the incubation period (236, 237). In certains cases, antibiotics
must be combined with vaccination if the effect is to be satisfactory. It is
known, for example, that penicillin protects monkeys against pulmonary an
thrax, but that this protection is effective only as long as the treatment con
tinues (94) . If, on the other hand, the animal is inoculated during the anti
biotic treatment, then full protection may be obtained (238).
A successful BW attack will present all the problems of mass casualty
treatment, but differences in age, race, sex, stress, nutritional level, dosage,
relative immunity, synergism, or antagonism with regard to already estab
lished infectious diseases, total body irradiation and possibly chemoprophy
laxis, will all tend to distribute the cases along a time scale. This will permit a
certain measure of care to be provided by medically unqualified-or para
medical personnel such as pharmacists (226) . This possibility must always be
borne in mind since it is unlikely that normal medical facilities would mea
sure up to the needs following a major BW attack, particularly if this in
volved agents of which there was no past experience. The enemy might, of
course, also launch the attack at a time when the medical and public health
facilities had been disrupted by military action or saturated by refugees.
Needless to say, a nuclear attack would severely reduce medical capabil
ities, for instance, the possibilities of using attenuated vaccines (27) . The
accumulation of casualties after a chemical attack might also severely reduce
the possibilities of coping with a biological attack. However. it is by no means
certain that the entire population in the target area will become seriously ill,
and thus the importance of the sick being cared for by those who are well, the
less seriously ill looking after themselves, and those who are well being pro
tected against panic, cannot be overrated (239). The proper handling of the
mass communication media of TV and radio plays a central role in this con
The problem of person-to-person transmission of disease after a BW at
tack would probably be regarded in a different light than it would be in peace
time. I n the case of smallpox or plague, an attempt would be made to mini
mize the risk for unexposed susceptible individuals, but for most diseases iso
lation would probably not be regarded as a major factor. As far as the United
States is concerned, it is believed that the available disease control methods
would prevent a significant secondary spread, provided that conventional or
nuclear warfare had not disrupted the public health services (26) . Table I
lists some parameters which would govern the medical defence in a BW at




conta- Lethalityb

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Acidophuic Virus Diseases:

Adenovirus infections
Albo virus infections
Chikungunya fever
Dengue fever
Eastern e uine ence halitis
Japanese -encepha tis
Murray Valley encephalitis
Rift Valley fever
RSSE complex
Venezuelan equine encephalitis
Western equine encephalitis
West Nile fever
Yellow fever
B-virus infection
Po iomye itis






< 1- 1 0





+ ++











3-6-1 0
10-2 1




Psittacosis (oruithosis)




Boutonneuse fever
Rocky Mountain Spotted fever


Basophtlic Virus Diseases:

Rickellsial Diseases:



Scrub typhus
Siberian typhus
hus (epidemic)
o hyuian fever

Bacterial Diseases:

Anthrax (pulmonary)





2- 10


++ +





Plague (pulmonary)

Fungal Diseases:

Protosoan Diseases:
Malaria (Plasm<Jdium jalciparum)











Z-4-1 4
1-4-8-2 1




+++ 100
++ +












Routine rOPhylaxiS
and t erap









CA?, TC?






TC, ( +)S


P, TC?







a The estimation of contagiousity is based on the assumption of overcrowding and primitive conditions,
+++ considerable, ++ not serious, + small or none. b Limits based on known outbreaks in nonvaccinated
populations not treated with ant b otiC chemotherapeutics, or sera. 0 Limits based on known outbreaks with
the commonly accepted times italicize . d Ant biotics listed in order of preference: AMPH-B =Amphotericine B, CA = Chloramphenicol, CH = Chloroquine, etc., N =Neosalvarsane, P =peuicillin, PABA =p-Aminobenzoic acid, S =Streptomycin, SU =Sulphonamides, TC =Tetracycline.



A m aj or debatable point is whether the gravest danger of BW lies in the

direct effects on the population or in the undermining of the economy by the
destruction of crops (240, 241) and livestock (242, 243). An enemy might use
a spectrum of plant viruses or any of the animal viruses which give rise to

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diseases such as African swine fever, hog cholera, Rift Valley fever, rinder
pest, foot-and-mouth disease, fowl plague, and Newcastle disease, or he
might employ the bacteria which give rise to anthrax, brucellosis, or glanders
(145). The problems of type and subtype identification for vaccine produc
tion can be illustrated by foot-and-mouth disease, which offers considerable
difficulties even in peacetime.
The defence measures required range from crop destruction and large
scale slaughtering to insect control and specific treatment, so they could be
covered only in a handbook. I n general, much can be gained by having stand
by facilities for the preservation of food and fodder, based on material from
diseased animals and plants.
The strain on the personnel resources would be considerable, as indicated
by the USSR plans for protection of cattle against ABC warfare (244). This
presupposes a need of seven men per one hundred animals. I n peacetime, a
special organization prepares different protection measures, stocks vaccines
and disinfectants, and executes a training program. Where an urgent readi
ness is required, the cattle are transported to the safest possible location. I n
the case of a biological attack special measures such as quarantine and de
contamination are taken, depending on the agent and the method of spread

I n the protection of a nation's water resources and its food and drug pro
duction against BW, security checks and special training of all personnel hold
ing key positions in sabotage-vulnerable establishments are essential ( 143,
245). Food processing industries, dairies, ice cream factories, and waterworks
are at the head of a long list of vulnerable establishments.
Considering modern water purification methods and the close supervision
of the water supply which is the rule with military units, the water contami
nation problem is primarily one for civilian authorities to consider (246). Con
taminated water should be filtered, boiled, or exposed to high-grade chlori

nation in accordance with standard handbook procedures. However, other

methods of treatment may also have some value (247).

The exposure of man's environment to a biological weapon may have fat

reaching ecological and epidemiological effects which require attention. Par
ticularly, some viruses such as rabies and the normally vector-borne infecti
ous agents may attack a number of unrelated species. The latter are gener
ally not as specific as the microorganisms which normally gain entrance via
the respiratory tract. D. D. Fothergill has emphasized our lack of knowledge
of the susceptibility of many of the mammals, birds, reptiles, amphibia, and
insects that would be exposed during an aerosol attack, and he has raised a
number of very important questions (248): "Would new and unusual zoonot-



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ic foci of endemic disease be established? Would it create the basis for pos
sible genetic evolution of agents in new directions with changes in virulence
for some species? Would i t establish some post-attack public health and en
vironmental problems that are unique and beyond our present experience?"
Bearing such questions in mind, it is certainly wise to be prepared for control
measures (for example, mosquito destruction) aimed at restoring the normal
ecological balance. Without such possi bilities, even the most perfect physical
protection of man would be inadequate, since he might become a victim of
newly established disease reservoirs.

Surveillance of communicable diseases. Survei ll a nce means the epidemio

study of a disease as a dynamic process involving the ecology of the
infectious agents, the host, the reservoirs, and the vectors, as well as the com
plex mechanisms concerned in the spread of infection and the extent to which
this spread occurs (233) . On the one hand, it requires the systematic collec
tion of morbidity and mortality reports, together with l aboratory data, and
on the other, it i nvolves the analysis, interpretati on, and distribution of the

l ogi cal

information received.

Since a proper surveillance program must permit an early recognition of

pattern and vector di stribution, it is fundamental to
any BW defence system. It should provide a solid base for the planning of
control measures such as vaccination, and for an optimum utilization of the
therapeutic resources. The speed required in BW defence is such that it may
necessitate such highly sophisticated approaches as the Command Automatic
Data Processing System (CADPS) used by the United States Army (137).
This provides an accounting system which reveals the medical situation at
any given moment.
Surveillance programs will vary depending upon local conditions, and
ch ang es in the d iseas e

they should naturally be aimed at guarding the most vulnerable areas.

Clearly, the vast oriental cities, where the water supply and environmental

cannot keep up with the rapid population growth, face differ

ent problems than those caused by the controlled expansion of the western
type megalopolis. In both locations, political, social, and economic factors
change the ecological situation, but in different ways, and this emphasizes
the importance of local surveys in order to evaluate the vulnerability to differ
health facilities

ent types of BW attack.

Most national surveillance programs lean heavily on the international ac

tivity of the World H ealth Organization (WHO) which, in 1965 , established
an Epidemiological Surveillance Unit in its Division of Communicable Dis
eases. The organization has had long experience with quarantinable diseases
(cholera, plague, relapsing fever, smallpox, typhus, and yellow fever) , in
which control measures are normally based on a combination of the results of
rapid laboratory diagnosis with the data from immunological and vector sur-

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veys, factors which are also significant in BW defence. Surveillance is also a

feature of the work done by the WHO I nfluenza Reference Laboratories, and
it is an integral part of the organization's campaigns against malaria and en
demic treponematoses.
National surveillance programs like that of the Communicable Disease
Center of Atlanta in Georgia (249), add information on different diseases to
the global picture (poliomyelitis, infectious hepatitis, influenza, salmonello
sis, shigellosis, diphtheria, etc.), but the data from many developing areas of
the world are incomplete. Surveillance of plague on the basis of a systematic
study of the ecology of the disease has, for instance, been carried out in Soviet
Russia, the United States, and Mainland China, but in several other coun
tries with natural foci a long-term systematic study does not exist (233). I t is
natural that such a lack may delay the detection of changes in the disease
pattern indicative of a BW attack. In a technically advanced society, on the
other hand, the normal surveillance of communicable diseases simplifies the
detection of such an attack.
Obviously, a systematic study of the spread of infections in a population
and a survey of certain diseases such as tularemia and Q-fever among ani
mals-as carried out at the I nstitute of Epidemiology and Microbiology in
Prague (250)-must provide a useful basis for BW defence planning. A na
tional surveillance program involving routine immunological testing and
regular surveys of vector populations provides a basis for retrospective in
vestigations which may constitute important proofs to support a case of BW
allegation, which is not an easy matter (66, 251). Particularly if supported by
automatic devices (which would not necessarily have to be as rapid as those
mentioned under the heading "early warning") it might reduce the risk of an
attack being launched. Cooperation between countries that aim at improv
ing surveillance methodology, might-together with improved WHO facili
ties-in fact, constitute a very powerful protection against insidious attacks
(252, 253). M ultipurpose immunological surveys are of particular significance
in this connection because, if based on standardized techniques, they "per
mit much better comparisons of the epidemiological situation among popu
lations living in different environmental conditions, than do existing morbid
ity and mortality statistics collected in areas with widely differing facilities
for reporting and varied criteria of diagnosis and accuracy" (233). To be of
real significance, such surveys should be broad and trans-national and they
would require advanced planning and data handling, facilities for automated
analysis and other facilities frequently beyond the reach of small nations.
However, the latter have not so far entrusted the WHO with special powers
and responsibilities in the BW area, a strange fact if one considers that their
own health services are far from adequate in such a connection. Those ser
vices might help to ward off epidemics which normally respond to quarantine
measures, but they could hardly arrest the drift of an aerosol cloud or the
spread of arthropod vectors from a massive release. The effects of such at-

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tacks might reach far beyond national boundaries and effective limitation
might require efforts of a magnitude which would exceed the capabilities of
many nations. Such countries should be able to buy an "insurance policy"
which would guarantee immediate airmail deliveries of substantial quantities
of vaccines and antibiotics in case of attack, beside providing personnel and
advice (252).
Several of the United Nations specialized agencies have a certain compe
tence in the BW defence area, however. As mentioned above, WHO can be
expected to have an interest in the public health aspects but, in addition, the
Food and Agricultural Organization should be concerned with plant-animal
protection, and UNESCO in the associated basic microbiological research,
documentation, and science policy. From this point of view but also consid
ering the political dangers of impromptu appointments of international com
mittees to study cases of BW allegations, the establishment of an indepen
dent International Microbiological Agency as a parallel to the I nternational
Atomic Energy Agency (IAEA) has also been considered (254) . This would
tie a control function in BW to the peaceful applications of microbiology, in
the same way as the control of reactor fuels is tied to the peaceful uses of
atomic energy within the IAEA.
Planning of biological warfare defence.-I n addition to an effective system
for a nation-wide reporting of infectious diseases, the planning of BW
defence should involve the preparation of stand-by legislation for compulsory
immunization, stockpiling of therapeutics, preparations for extra produc
tion of broad-spectrum antibiotics, microbiological research and, finally, the
spread of knowledge about unusual infections (52) . The administrative mea
sures and the training of personnel such as police, customs officers, etc., must
vary from country to country, but Sweden can be chosen as an illustration of
the use of an approadi which is natural to a small country (255-257).
There is, in Sweden, a commission on medical catastrophes which comes
under the Department of Health, and which coordinates all activities re
quired by maj or fires and traffic accidents, explosions, accidental release of
chemicals, nature catastrophes, etc. This commission is also responsible for
the necessary emergency measures to be taken against major epidemics as
well as against poisioning via air, water, and food. The production of detailed
action plans, on the other hand, is the responsibility of special regional and
hospital committees. These committees prepare detailed catalogues of mea
sures to be taken in a number of conceivable situations. The regional com
mittees consider matters like sounding the alarm, the distribution of cases
among available treatment facilities, internal communications, possible iso
lation of the stricken area, providing it with personnel and transport facil
ities, etc. The hospital committees, on their part, plan for the 1Llarming of
hospital personnel, for the distribution of responsibilities, for the use of med
ical supplies, for information to relatives, press, and radio, for the relocation
of patients under care before the emergency occurred, and, finally, for the
registration, sorting, and treatment of mass casualties.




The inhibitory forces which influence biological warfare development.-Why

is it that biological weapons which would leave the material resources of the
attacked country undamaged and even spare the production potential, have
not been used on a large scale? This is a very difficult question to answer, but
the following points may be considered :
that critical problems in the production, storage, and delivery of some
agents, judged necessary to make up a fully operational system, have
not yet been solved ;
2. that the attacking country's own BW defence is not adequate or has
not been fully tested.
The existence of a sophisticated aerosol warning system in a country
might not be enough to induce that country to use infectious aerosols
offensively, since this would invite biological sabotage acts which would
circumvent that particular type of defence;
3. that the popular conception of biological weapons as terror agents
makes their use most distasteful, at least in countries with a free press
and a democratic system of government;
4. that the value of such weapons is relatively less to the nuclear armed
powers, which have a great BW development potential, than to the
smaller nations which have a limited BW capability ;
5 . that all of the tactical/strategic implications, for example, the oppor
tunities for small groups to bring about devastating reprisals, have not
yet been given serious attention in military-political discussions;
6. that the exact target area and the long-range ecological consequences
are very difficult and occasionally impossible to predict ;
7. that there exists a sort of pactum turpae based upon the unpredictability
and complicated consequences of BW. These must introduce very dis
turbing elements in the mathematical mode of military thinking.

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How long a period of grace may last when it rests on as complex a founda
tion as this, is impossible to tell, but the psychological factors which underlie
the Geneva protocol of 1925 still seem to exert a strong inhibitory influence.
However, the time available for talks about BW disarmament may not be
very long because one should not disregard the forces which give substance to
such remarks as the following: "The mores of a society vary with the degree
to which its existence is threatened" (58), and "frustration and a sense of
futility can make even desperate measures seem attractive. What is 'un
thinkable' at one moment may be policy the next" (12).
The case for disarmament.-A rewarding aspect of BW defence is that it
will improve the possibilities of coping with naturally occurring diseases.
Where those possibilities are already good and budget considerations do
not set a ceiling that is too low, the defence problems may not be insoluble
"if approached unemotionally, with adequate knowledge and sound reason
ing" (58) . Whether an ultimate state of nation-wide, semicontinuous protec-

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tion will ever become acceptable from the social and medical points of view is,
however, another matter. In the developing parts of the world and, in some
smaller countries, on the other hand, the outlook is less "bright" and multi
lateral or international collaboration might be required (252) . In such areas
the risks are probably greater also, because the building of a nuclear capabil
ity is apt to be a slow process, and biological weapons might offer a tempting
strategic alternative. Since this could well upset the delicate power balance
which now affords a precarious but fundamentally important stability in the
world, the superpowers might have a very real interest in BW disarmament
and contro!' They should be in a position to know that the biological weap
ons are likely to remain erratic and difficult to handle as part of military
games theory, the more so, the cruder the technology. One simulated attack
has, for instance, been claimed to have killed or incapacitated some 600,000
friendly or neutral civilians at the same time that 75 per cent of the opposing
troops were eliminated (51). Finally, the superpowers should know that a
fully effective defence program will i nvolve almost prohibitive costs and con
stitute a drain on a professional group which has many critical functions in
society and might also be of considerable significance for the aid programs
which are now an important part of their foreign policy.
Since the power balance seems to offer the only available road toward a
distant goal of world order preserved by some sort of world government, a
disarmament agreement should also be acceptable to such smaller nations
that have reached a certain state of maturity. When a responsible superna
tional authority has eventually been established (258), there might be reason
to reconsider the situation (3 1). This is because-as soon as the risk of escala
tion is eliminated (see below)-the use of incapacitating biological weapons
in previously mass-vaccinated areas might offer the ultimate answer to the
control of subversive infiltration and to certain other problems with which a
world government might find itself confronted.
Like most political problems, the matter of BW disarmament is a multi
factorial equation, in which the distribution of attitudes toward the weapons
enters as a very important element. These attitudes range from efforts to
classify BW weapons as the most humane among arms, to condemnations
full of repugnance and moral indignation. As long as we accept conventional
methods of warfare, there is obviously no simple answer.
Discussions on the relative moral merits of napalm burning, saturation
bombing, and nuclear warfare versus the use of infectious aerosols actually
seem pointless without a careful balancing of the long-range interests in terms
of self-expression and happiness, both as far as the attacker and the attacked
are concerned. Such a type of penetrating analysis is difficult for anybody
and particularly for the military man and the average politician. Superfici
ally, the biological weapons might seem preferable, because they include a
wide range of incapacitating agents [for instance, Rift Valley fever (31)], and
they do not have the obvious genetic effects that we associate with nuclear

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weapons. However, this is hardly enough to open the door to their use, since
the young, the elderly, and the infirm may become fatalities and the range
proceeds further to include agents of high mortality [for instance, A ctinobacil
lus mallei or botulinum toxin (31)1. Large-scale deployment of viruses might
also, according to J. Lederberg (259) , be a potential threat against the whole
species as uncontrollable mutant forms could well develop. Once the use, of
biological weapons became acceptable, the " human" aspects would also be
offset by an almost inevitable escalation. In fact, this is the strongest argu
ment for disarmament and for control ill the BW area ( 122) , an argument
that would seem to cancel out even the seemingly most acceptable reasons
for developing offensive biological weapons. Among the arguments for BW
disarmament is the one that must also emphasize the relatively unselective
effects of biological weapons on large-target populations.
Until Homo sapiens develops a global conscience, men will obviously con
tinue to die believing that they fight for freedom from want and freedom from
fear. The hunger and fear of the 20th Century seems indeed to be a mockery
of this belief, and all educated adults who accept the growing gap between
rich and poor nations with indifference, take a share in the responsibility for
the conflicts which now seem inevitable. Such conflicts might well breed wea
pons which are copied after Nature's own ecological system. Man's success
with biological control of pests and insects indicates a road which is certainly
appalling, although not necessarily because biological agents are invisible and
lack smell and taste, or because a perfect defence will never exist except in
the minds of some theoreticians. What determines the attitude of the author
is rather the fact that those weapons will never be selective enough to spare
the individuals who are not responsible for the situation which breeds them.
As M orris West has put it (260) : "A child has no politics. A child has no
nationality. He has only the right to live, the right to hope. If these rights
are denied him, it is a crime against humanity, and every honest man must
raise his voice against it." The same argument can, of course, be used against
any of the modern mass destruction weapons, but there is a very special rea
son against the development of biological weapons. It is that they come from
some of the largest and best operated establishments for applied microbio
logy in the world of today and, unlike other military establishments, they
would'require no major reorganization in order to switch most of their activ
ities to a highly constructive path, where the first steps have in fact already
been taken (261) . The potential of military microbiology is illustrated by its
civil "spin-off" : vaccines against Newcastle disease, fowl plague, and rinder
pest, improved diagnostic and therapeutic methods, and so on (2, 15),
The American center for BW research, Fort Detrick, near Fredrick in
M aryland, has a scientific staff of 120 Ph.D's, 1 10 M .Sc's, 320 B .Sc's, 34
D.V. M's and 14 M . D's. It occupies 1300 acres of land and is housed in a
building complex valued at $75,000,000 ( 1 1 ) . This indicates that applied
microbiology would probably gain much by a shift in emphasis. As far as

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Britian is concerned, it has been suggested that the military Microbiological

Research Establishment at Porton should be charged with functions in in
dustrial microbiology (262) .
" Public Health could be Biological Warfare in reverse" (58) , because
most of the efforts which go into the development of weapons could equally
well be applied to the development of new vaccines, new administration tech
niques, and other methods relevant to disease prevention, which is such an
important factor in the industrialization process of the developing countries.
It is true that BW in reverse might tend to accelerate the "population explo
sion" but it could also provide new answers to some problems in family plan
ning (d. immunological control of sperm production) . To deny its potential
would also be to deny the impact of antibiotics development on public health
and husbandry, of biological insect control and seed inoculation on agricul
ture, and of the fermentation technology and food microbiology on the pro
duction of nutrients. The conference : "Global Impacts of Applied Microbio
logy," held in Stockholm in 1963, served as a reminder of the significance of
applied microbiology to the developing countries of the world (263, 264).
Realizing this, UNESCO has launched an ambitious microbiological program
and a number of individual countries have included microbiological efforts
in their aid programs, If their significance can be conclusively proven, and if
the desire to benefit becomes strong enough in the developing countries, they
might, perhaps, be indu ced to accept BW control and inspection as a match
ing contribution toward a masterplan for peaceful development. After all,
the International Atomic Energy Agency requires control of the fuel passing
through the reactors which it sets up.
Control and inspection problems.- Different types of nonproliferation trea
ties in the BW area have been proposed (259, 265) but, in the postwar period,
the United States has become increasingly unwilling to accept treaties re
stricting armaments if they do not embody the principle of adequate inspec
tion. With regard to biological and chemical weapons, the official attitude
has been that effective controls on biological and chemical weapons " may
have to await international agreements with necessary safeguards" (5) .
There is, however, a consensus of opinion, expressed in the Soviet and West
ern proposals for general and complete disarmament to the effect that biolog
ical and chemical weapons should eventually be subject to control. So far,
one has not, however, " come to grips with the formidable technical problems
which would accompany an agreement to reduce or prohibit biological and
chemical weapons" (5) . To insure against the delivery of biological weapons
in a disarmed world might prove to be impossible, owing to the many delivery
systems available to the potential user. I nspection including registration of
scientific personnel, accounting of materials, fiscal controls, and visits to con
ceivable production and testing facilities also involves extremely difficult
problems (266) . However, if an efficient system for the regulation of research
and development and of methods for preventing testing and clandestine
stockpiling cannot be proposed, it is not likely that nations wiII be prepared

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to disarm. The Pugwash Organization early recognized this problem (267)

and in recent years has operated a special BW study group to analyze the
matter. A proposal on the limitation of biological weapons development (265)
and reports on the associated control problems (268) and documentation
needs (269) have appeared, as have papers on the delineation between offen
sive and defensive research in the BW area (270) , on declassification (27 1),
and on the means available to guard against the military application of ncwly
acquired microbiological knowledge (272) . Finally, the Pugwash study group
has reported on BW protection through international cooperation (252) , on
inspection for allegation (253) , and on the techniques useful in the latter
connection (273) . A series of experimental i nspections have also been per
formed in four countries in order to gain practical experience (274) . This
study has not yet been concluded, but the inspectors have come to the con
clusion that the possibilities of arriving at a reasonably effective control sys
tcm are bctter than originally cxpected (254, 275) . Finally, the International
I nstitute for Peace and Conflict Research (SI PRI) has initiated feasibility
studies dealing with early warning devices (120, 1 24) and with methods for
the detection of remote biological weapons testing (276) .
Control and inspection i n the BW area i s hardly possible with regard to
"sabotage" quantities, but large-scale military efforts are probably not easy
to hide. For instance, the Pine Bluff Arsenal i n Arkansas where biological
weapons (31) are produced, as well as toxic-chemical and riot control muni
tions, covers some 1 5 ,000 acres (12). Facilities for field testing would seem to
be even more difficult to conceal. The Dugway Proving Ground is an example
occupying an area in Utah larger than the state of Rhode Island (12). A
high intensity solar irradiation might help to accelerate the biological decay
in such a location (277) but infected animals might function as revealing dis
ease reservoirs (278) and some microorganisms might be so resistant that
they would be detectable by normal sampling techniques over long periods
and at considerable distances from the test site (276) . A high degree of re
sistence has, for instance, been observed in the case of anthrax, which was
used for BW experiments on the small island of Gruinard, off the northwest
coast of Scotland, during World War I I . After a recent study, Dr. G. E. Gor
don Smith stated that this island "may remain infected for 100 years" ( 1 2) .
However, the control and inspection problems are staggering because
there is nothing about a production facility which would necessarily show on
the outside of the buildings, and extensive field tests might be staged covertly
on foreign soil. Consequently, an intelligence activity would have to be part
of the effort (269) . To be fully effective, unorthodox supporting initiatives
might also be required such as, for instance, internationalization of the mi
crobiological profession (269) or providing diplomatic immunity and awards
for disclosure of military preparations contrary to agreement (279) . From
the political point of view, however, it might not be necessary to have an ab
solutely "leakproof" system. The most important aspects might be found in
side effects. The preparations for a control and inspection system would re-

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quire contacts that would help to reduce the mistrust among nations and
they would tend to reinforce the international conscience in the field (122).
International and regional agreements aimed at neutralizing the political
dangers of BW allegations and at limiting the spread of weapons technology
would then logically follow. I n such a process statesmen would be exposed to
the u npleasant facts of human ecology, and they would also receive a forceful
reminder of the fact that war is rapidly becoming obsolete as a means of
solving i nternational disputes. The political ideas arising at the turn of the
century when masses of people were regarded as a military advantage, do not
ring true in the age of "megacorps" and "overkilL" It is sad that we must
live with such terms until we learn to think in global terms as far as man's
human and material resources are concerned.

The author is indebted to Dr H. Markkula of the Swedish Defence Re

search Establishment (FOA) for the material summarized in Table I , and for
several texts which are not easily accessible. He has also given much helpful
advice and contributed constructive criticism. The Swedish Civil Defence
Medical Director, Dr W. von Greyerz, Dr R. Bj ornerstedt of the I nterna
tional Peace Research Institute (SIPRI), and the members of the Executive
Committee of the Swedish Microbiological Society have also shown a stimu
lating interest and made a number of valuable comments.



1. Editorial,


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169, 1 p. (1966)



2. Brown, M. H., Can. Med. Assoc. J.,

67, 543-49 (1952)
3. Noel-Baker, P., Impact, 15, 2 1 1-46
4. Stubbs, M., Advan. Chem. Ser., 26,
34-40 ( 1960)
5. Pfal tzgraff, R. L., Jr., Current History,
4 7, 1 8-24, 5 1 ( 1964)
6. Rosebury, T. Biological warfare and
disarmament. Paper presented at a
forum under the auspices of The
Society for Social Responsibility
in Science. (Sanders Theatre, Har
vard Univ., Boston, 1960)
7. Coggins, C. H., Armed Forces Chem.
J., 19" 3, 7-8, 10-12 (1963)
8. USSR Ministry of Health, Military
Epidemiology. (in Russian) (Rogo
zin, I. I., Ed., Medgis, 135 pp.,
9. Goun" L., Civil defense in the Soviet
Union. (Univ. of California Press,
Berkeley-Los Angeles, 207 pp.,
10. U.S. House of Representatives, Com
mittee on Appropriations. Depart
ment of Defense Appropriation for
1962. 87th Congr., 1st Sess. Hear
ings. (Govt. Printing Office, Wash
ington, D.C., 1961)
1 1 . Langer, E. , Science, 155, 1 74-79 (1967)
1 2 . Langer, E . , Science, 155, 299-303
13. Glassmann, H. N., Bacterial. Rev.,
30, 657-59 (1966)
14. Krueger, A. P., Military Surg., 405-18
15. Merck, G. W., Bull. A tomic Sci., 2,
1 6- 1 8 (1946)
1 6. Dixon, B., New Scientist, 32, 558-59
17. Anonymous,
(Ontario) aune 6, 1962)
18. Rabi, 1., New York Times (January
I, 1958)
19. Wedum, A. G., Bacterial. Rev., 25,
210-16 (1961)
20. Briggs Phillips, G., Microbiological
safety in U.S. and foreign laborator
ies, Tech. Study 35. (U.S. Army
Chern. Corps Res. and Develop.
Command, Fort Detrick, Md., 291
pp., 1961)
21. Edwards, R. W., Favero, M . S., Hoff

man, R. K., Lanahan, T. B.,


Leod , N. H., MCDade, J. J.,

Skaliy, P., Briggs Phillips, G., A

selected bibliography on microbiolog

ical contamination control in the de
sign of laboratories. (Am. Assoc. for
Cont, Control, 10 pp., March, 1965)
22. Edwards, R. W., Favero, M. S., Hoff
man, R. K., Lanahan, T. B., Mac
Leod, N. H., M cDade, J. J" Skaliy,
P., Briggs Phillips, G., Microbi
ological contamination control. A
state of the art report. (Am. Assoc.
for Cont, Control, 1 7 pp., April,
23. Briggs Phillips, G., Jemski, J. V.,
Microbiological safety bibliography,
Miscellaneous Publ. 6. (U.S. Army
Biol. Labs. , Fort Detrick, 89 pp.,
July, 1965)
24. Environmental hygiene, Hoff, E . C.,
Ed. In M edical Department of the
U.S. Army. Preventive Medicine in
World WadI, 2. (Office of the Sur
geon General., Dept. of the Army,
Washington, D.C., 1955)
25. Communicable diseases transmitted
through respiratory and alimentary
tracts. Hoff, E. C., Ed., In Medical
Department of the U.S. Army.
Preventive Medicine in World War
II, 4. (Office of the Surgeon Gen
eral ., Dept. of the Army, Washing
ton, D . C . , 1958)
26. Crozier, D., Military Med., 128, 8 185 (1963)
27. Raska, K., Havlik, 0., Chladek, V.,
Novotny, J., Pi'ivora, M., Symon,
K., Weiser, J . , Wolf, A., Dey Ge
Krieg. (VEB Verlag Volk und
Gesundheit, Berlin, 1 64 pp., 1962)
28. Rosebury, T., Bull. Atomic Sci., 16,
22 7-36 (1960)
29. ACS Board of Directors Special Com
mittee on Civil Defense, Chem. Eng.
News, 37, 76-80 (1959)
30. Ronneberg, C. E., Advan. Chem. Ser. ,
26, 1-7 (1960)
3 1 . Rotschild, J. H., Tomorrow's Weapons
Chemical and Biological. (M cGraw
Hill Book Co., New York-Toronto
London, 2 7 1 pp., 1964)
32. Editorial, Army, 14, 37-39, 42 (1964)
33. Ley, H. L., Jr., Smadel, J. E., Diercks,
F. H . , Paterson, P. Y. A m . J.
Hyg., 56, 3 1 3-18 (1952)
34. Boyd, M . F., Kitchen, S. F., Am J.
Trop. Med. Hygs., 23, 209-25 (1943)
35. Morsles-Otero, P., J. Public Health
Trap. Med., 6, 3-88 (1930)
36. Shaughnessy, H. L., Olsson, R. C . ,


Bass, K., Friewer, F., Levinson,
S. 0., J. Am Med. Assoc., 132,
362-68 (1946)

Annu. Rev. Microbiol. 1967.21:639-676. Downloaded from

Access provided by Universidade Federal de Alagoas on 01/02/16. For personal use only.

37. M cColl ough , W B., Eisele, C. W., J.

Infect. Diseases, 88, 209, 259, 2 78

38. Koprowski , H., Jervis, G. A., Morton,
T. W., Am. J. Hyg., 55, 108-26
39. Saslow, S., Wilson, H. E., Prior, J. A.,
Carhart, S., Centro Soc. Clin. Res.
(Chicago, IlL, Nov 5, 1959)
40. McCrumb, F. R., Jr. , Bacterio!. Rev.,
25, 262-67 (1961)

in : Fiirsvars- och katastrofmedicin.

beredskapsnamnd, Stockholm, 485
pp., 1966)
5 7 . Rich, F., F. A. S. Newsletter, 1 (June,

58. Tigertt, W. D., Military Med., 128,

135-44 (1963)
59. Davis, C. N., Inhaled Particles and
Vapours. (Pergamon Press, Ltd.,
Oxford, 495 pp., 1961)
60. Riley, R. L., O'Grady, F., A irborne

41. Tigertt, W. D., Benenson, A. S.,

GOChenour, W. S., Bacteriol. Rev.,


42. Tigertt, W. D., Benenson, A. S.,

Trans. Assoc. Am. Physicians, 69,


25, 285-93 (1961)

98-104 (1956)

43. Fothergill, L. D., Proc. Conf. on Med.

Civil Defense. Co uncil on Natl.
Defense. (Am. Med. Assoc., San


Francisco, Calif., June 2 1 , 1958)

44. Fothergill , L. D., A dvan. Chem. Ser.,

26, 23-33 (1960)
45. Fothergill, L. D., New Scientist, 263,
546-47 (1961)
46. Wright, G. W., Bacteriol. Rev., 25,


2 19-27 (1961)

47. Hatch, T. F., Bacteriol. Rev., 25, 23740 (1961)

48. Wells, W. F., A irborne Contagion and
A ir Hygiene. (Harvard Univ. Press,
Cambridge, 423 pp., 1955)
49. Wolfe, E. K., Jr., Bacteriol. Rev., 25,
194-202 (1961)

50. U.S. House of Representatives Com

mittee on Science and Astronautics.

Biological, and Radio

logical Warfare Agents,

Congr., 1st Sess., Hearings. (Govt.
Printing Office, Washington, D.C.,

51. Schneir. W., The Reporter, 24-28.

(October I, 1959)
5 2. Ha rtford , T. J., Military Med.; 128,

1 45-46 (1963)
53. Civil De/ense A gainst Biological War
fare. Fed Civil Defense Adm. Tech.
Manual TM 1 1-10. (Govt. Print

ing Office, Washington, D.C., 1953)

54. Biological and Chemical Ware/are.
Condensed papers Pugwash meet
ing 1959 and ACS Symposium
1960. Bull. A tomic Sci., 16, 226-53

Defense against biological warfare-A

Symposium, Military Med., ;128
81-144 (1963)
56. Markkula, H., Biological weapons and
their effects (in Swedish), 205-37,

Transmission and Con

trol. (Macmillan Co., New York,

180 pp., 1961)
Conference on Airborne Infection,
Miami Beach, Fla., 1960, Bacterial.
Rev., 25, 1 7 3-382 (1961)
Proc. Intern. Symp. A erobiology, 1st,
Berkeley, Calif., 1 963. (Naval BioI.
Lab., Naval Supply Center, Oak
land, Calif., 405 pp., 1963)
International Conference on Aero
biology, 2nd, Chicago, 1 966. Air
borne Infection, Bacteriol Rev., 30,
485-698 (1966)
The 1959 Fort Detrick Symposium
on Nonspecific Resistance to In
fection. (Hood College, Frederick,
Maryland, 1959), Bacteriol. Rev.,
24, 1-200 (1960)

65. Materials on the trial 0/ former service

men of the Japanese A rmy charged
with manufacturing and employing
bacteriological weapons.
Languages PubJ. House, Moscow,

535 pp., 1950)

Commission for the investigation of
the facts concerning bacterial war
fare in Korea and China. (Peking,
665 pp., 1 9 5 2)
67. Jenkins, D. W., Military Med., 128
1 1 6-18 (1963)
68. Finlay, C. J., Carlos Finlay and Yellow
Fever. (Oxford Uni". Press, 261 pp.,
69. Boyd, M. F., Malariology, Compre
66. Report of the International

hensive Sur1Jey of all A spects of this

Group of Diseases from a Global
Standpoint, I, 2 CW. B. Saunders
Co., Philadelphia, 1643 pp., 1949)

Simmons, J. S., St. John, J. H.,

Reynolds, F. H . K., E"perimental
studies of dengue. Monograph 29
(Philippine Bur. Science, 1931)
71. Lumley, G. F., Taylor, F. H., Dengue,
Pts. 1 & 2., Service PubJ. 3., School
Publ. Health and Trop. Med.
(Univ. of Sidney, Commonwealth
Dept. of Health, 173 pp., 1943)


72. Proctor, D . F., Bacteriol. Rev., 30,
498-51 3 (1966)
73. Landahl, H. D., Black, S., J. Ind.
Hyg. Toxicol., 29, 269-77 (1947)
74. Sawyer, W. D., Military Med., 128,
90-93 (1963)
75. Couch, R. B., Cate, T. R., D ouglas,
R. G., Jr., Gerone, P. J., Knight.
V., Bacteriol. Rev. , 30, 5 1 7-29


R. J., Leonard, F. A.,

Bacterial. Rev., 25, IS2-S7 (1961)
77. Brown, J. S., Cook, K. M . , Ney, F. G.,
Hatch, T., Am. J. Public Health, 40,

Annu. Rev. Microbiol. 1967.21:639-676. Downloaded from

Access provided by Universidade Federal de Alagoas on 01/02/16. For personal use only.

76. Goodlow,

450-5S (1950)
7S. Landahl, H. D., BUll. Math. Biophys.,
12, 43-56 (1950)
79. Landahl, H . D., Tracewell, T. N.,

Lassen, W . H.,




Arch. Ind. HyC.

Toxieol. , 3, 359-66 (1951)
Mitchell, R. 1., Am. Rev. Respirat. Dis
eases, 82, 627-39 (1960)
Kass, E. H . , Green, G. M . , Goldstein,
E . , Bacteriol. Rev., 30, 488-96 (1966)
White, J. D., [Personal communica
tion to W. D. Sayer, Ref. (74)]
McGavran, M . H., Beard, C. W.,
Berendt, R. F., Nakamura, R. M.,
Am. J. Pathol., 40, 653-70 (1962)
Oren, R., Cited in Medical and BioI.
Research in Israel. (Thesis. Prywes,
M . , Ed., The Hebrew University
of Jersualem, 1960)
Buescher, E. L., Bellanti, J. A., Bac
teriol. Rev., 30, 539-41 (1966)
Benenson, A. S., Military Med., 128,

1 19-28 (1963)

87. Grimenez, D . F., Ciccarelli, A . S., A

new type of Clostridium botulinum,
Proc. Intern. Symp. Food Microbiol.,
5th Moscaw, July, 1966 (In press)
88. Belton, F. C., Henderson, D. W.,
Brit. J. Exptl. Pathol., 37, 156-60

89. Thorne, C. B., Belton, F. C., J. Gen.
Microbiol., 17, 505-16 (1957)
90. Wright, G. G., Hedberg, M . A.,

Feinberg, R. J., J. Exptl. Med., 93,

523-27 (1951 )
9 1 . Saslaw, S . , Eigelsbach. H. T . , Prior,

J. A., Wilson, H. E., Carhart, S.,

Arch. Internal Med., 107, 702-14

92. Hornick, R. B., Eigelsbach, H. T.,
Bacteriol. Rev., 30, 532-38 (1966)
93. Benenson, A. S., Q-fever vaccine :
efficacy and present status , 47, in

Symp. on Q-fever. Med. Sci. Publ.,

A rmy Med. Servo Grad. School,
Walter Reed Army Med. Center, 6


94. Henderson, D. W., Peacock, S., Bel-


ton, F. C., J. Hyg., 54, 28-36 (1956)

95. Aleksandrov, N. 1., Gefen, N. Y.,
Voyenno Meditsinskiy Zh. No. 11

96. Aleksandrov, N. 1., Gefen, N. Y.,
Garin, N. S., Gapochko, K. G.,
I. 1., Sergeyev, V. M . ,
Voyenno Meditsinskiy Zh., 1 2 , 34-

Daal- Berg ,

38 (1958)
97. Eigelsbach, H. T., Tulis, J. J., Over
holt, E. L., Griffith, W. R., Bac
terial. Proc., 89 (1960)
98. Crozier, D., Woodward, T. E . , Mili
tary Med., 127, 701-5 (1962)
99. Eigelsbach, H. T., Tigertt, W. D.,
Saslaw, S., McCrumb, F. R., Jr.,
Proc. Army Science Conf. U.S.
Military Acad., West Point, N. Y.,

1, 235-46 (1962)
100. Tigertt, W. D., Bacteriol. Rev., 26,
35,4-73 (1962)
101. Lebidinsky, Y. A., Zh. Microbiol.,
Epidemiol., Immunobiol., 10, 14042 (1963)
102. Eigelsbach, H . T., Hornick, R. B.,
Occupational tularemia. Occupa
tional diseases acquired from ani
mals., 295-302, in Continued Educa
tional Ser. No. 124 (Univ. of Mich
igan, Ann Arbor, 1964)
103. Aleksandrov, N. I., Gefen, N. Y.,
Garin, N. S., Gapochko, K. G.,
Sergeyev, V. M., Smirnov, M. S.,
Tamarin, A. L., Shlyakhov, N. E.,
Voyenno Meditsinskiy Zh., 8, 27-32

104. Middlebrook, G., Bacteriol. Rev., 25,
331-46 (1961)
105. Dowling, H. F., Bacteriol. Rev., 30,
485-87 (1966)
106. Korostovtsev, S. B., Onikiyenko, B.
Khokhlov, L. 1.,
Meditsinskiy Zh., 3, 70-7 1 (1960)
107. Aleksandrov, N. I., Gefen, N. E.,
Gapochko, K. G., Garin, N. S.,
Sergeyev, V. M., Lazareva, E. S.,
Mishchenko, V. V., Shlyakhov, E.
N., Zh. Microbiol., Epidemiol. , Im
munobiol. USSR (Engl. transl.),
32, 1245-52 (1961)
lOS. Muromtsev, S. N., Borodiyuk, N. A.,
Nenashev, V. P., Zh. Microbiol.
(Engl. transl.) 3 1 , 803-6 (1960)
109. Aleksandrov, N. I., Gefen, N. E.,
Egorova, N. B., Sergeyev, V. M.,
Matyuk, P. D., Smirnov, M. S.,
Zh. Microbiol. Epidemiol. Immun
obiol. USSR. (Eng1. trans1.) 31,

1307-12 (1960)
1 10. Muromstev, S. N., Brodiyuk, Na A.,
Nenashev, V. P., Aleshina, R. M . ,


Zh. Microbiol. Epidemiol. Immun
obiol. USSR. (Eng!. trans!.) 32,
589-94 (1961)

Harris, M . M., Decker, H. M.,

Buchanan, L. M., Dahlgren, C. M.,
Sampling Microbiological Aerosols,

1 1 1.

Yamashiroya, H. M., Ehrlich, R.,

Magis, J. M., Bacteriol. Rev., 30,

U.S. Public HeaUh Servo Monograph,

60 (1953)

1 12.

Bartlema, H. C., Bacteriol. Rev., 30,

624-3 2 (1 96 6)


(1 9 59)


1 13. Aleksandrov, N. 1., Gefen, N. E.,

Garin, N. S., Gapochko, K. G., Zh.

Annu. Rev. Microbiol. 1967.21:639-676. Downloaded from

Access provided by Universidade Federal de Alagoas on 01/02/16. For personal use only.

Microbiol. Epidemiol. Immunobiol.

USSR (Eng!. trans!.) 3 1, 1833-40
( 1960)

1 14. Aleksandrov, N. 1., Gefen, N. Y.,

Garin, N. S., Gapochko, K. G.,
Mikitin, V. M., Antonova, A. A.,
Mishchenko, V. V., Voyenno Med
itsinskiy Zh., 12, (1960)

Kuehne, R. W., Sawyer, W. D.,

Gochenour, W. S., Jr., Am. J. Hyg.,
75, 347-50 (1962)
1 1 6. SaWyer, W. D., Kuehne, R W.,
Gochenour, W. S., Jr., Military


Med., 129, 1 040-43 (1964)



Tyler, M. E., Shipe, E. L., Painter,

R. B., Appl. Microbiol., 7, 355-62

1 1 7.




1 1 8.

Bartlema, H. c., Ned Milit. Geneesk.

Military Med., 128, 1 10-15 (1963)

Tijdschr., 7, 1 2 7-35 (1954)

1 19. Heden, C.-G., Tek. Vetenskap. Forsk.,
34, 55-67 (1963)
120. Tammelin, L. E., The technical basis

for studies of rapid detection of B

weapons. Working document to
the Inter. Inst. for Peace and Con
flict Res. (SIPRI) , Stockholm, Nov.

Schelling, T. C., Reciprocal Measures

for Arms Stabilization. The Strat
egy of World Order, 4 (Falk, R A.,
Mendlowitz, S. H., Eds., World
Law Fund)
122. Report of Study Group on Biological
Warfare, Proc. Pugwash Conf. Sci.,

World Affairs, 14th, 24, Venice,

Italy, A pril, (1965)
123. Cheronis, N. D., Submicrogram Ex
(Intersci. Pubis. ,
New York London, 351 pp., 1961)

124. Conf. on Rapid Detection Methods

for Airborne Bacteria and Viruses
The Intern. Inst. for Peace and
Conflict Res. (SIPRI), Stockholm,
Sept. (1966)
125. Collis, R T. H., New Scientist, 27,
2 7-29 (1965)


Henis, Y., Gould, J. R., Alexander,

M., Appl. Microbiol. , 14, 5 1 3-24


Guysen , J. M., Tipper, D . J., Strom

inger, J. L., Methods Enzymol., 8,


685-99 (1966)

128. Wolf, H. W., Skaliy, P., Hall, L. B . ,

130. Shipe, E. L., Tyler, M. E., Chapman,

D. N., Appl. Microbiol. , 7, 34954 (1959)

Tyler, M. E., Shipe, E. L., APPI. Mi

crobio!., 7, 337-49 (1959)
132. Batchelor, H. W., A dvan. Appl. Mi


crobial., 2, 3 1 -64 (1960)

133. Kethley, T. W., Proc. Intern. Symp.

A erobiology 1st, Berkeley, Calif.,
1963, 397-400. (The Naval Bio!.
Lab., Univ. of Calif., 405 pp., 1963)
134. Malligo, J. E., Idoine, L. S., A ppl.
Microbiol., 12, 32-36 (1964)
135. May, K. R, Bacteriol. Rev., 30, 55970 (1966)

136. Gerone, P. J., Couch, R. D., Keefer,

G. V., Douglas, R. G., Derren
bacher, E. B., Knight, V., Bacterial.
Rev., 30, 5 76-84 (1966)

Rapalski, A. J., Military Med., 128,

94-96 (1963)

138. Sulkin, S. E., Bacterial. Rev., 25, 2039 (1961)


Briggs Phillips, G., Causal Factors in

Microbiological Laboratory A cci
dents and Infections, Misc. Pub!. 2 .

(U.S. Army Bio!. Labs., Fort Det

rick, Md., 252 pp., 1965)
140. Lennette, E . H., Koprowski, H., J.
Am. Med.

A ssoc.,



Slepushkin, A. N., Vopr. Virusol., 4 :3,

3 1 1-14 (1959)
142. Shubladze, A. K., Gaydamovich, S. Y.,
Gavrilov, V. 1., Vopr. Virusol. , 4 :3 ,
305-10 (1959)
143. Beacham, L. M., Jr., Leininger, H. V.,

McConnell, H. J., Mathews, J. A.,

Spiher, A. T., Jr. Civil Defense In
formation for the Food and Drug
Officials. (U.S. Dept. of Health,
Educ. Welfare Food and Drug
Adm., Washington, D.C., 1956)
144. Emergency Manual Guide No HEW-2,

Effects of biological warfare agents.

(U.S. Dept. of Health, Educ. Wel
fare, Washington, D.C., 1 95 9)
145. Ronneberg, C. E., Chairman ACS
Committee on Civil Defense ; Non
military Defense, Chemical and
Biological Defenses in Perspective
A dvan. Chem. Ser., 26, (1960)

Anker, H. S., Bull. Atomic Sci., 16,

246-47 (1960)

Annu. Rev. Microbiol. 1967.21:639-676. Downloaded from

Access provided by Universidade Federal de Alagoas on 01/02/16. For personal use only.


147. Kaplan, M. M., Bull. A tomic Sci., 16,
23 7-40 (1960)
148. Albrink, W. S., Bacteriol. Rev., 25,
268-73 (1961)
149. Brachman, P. S., Kaufmann, A. F.,
Dalldorf, F. G., Bacteriol. Rev., 30,
646-57 (1966)
150. Meyer, K. F., Bacteriol. Rev., 25,
249-61 (1961)
1 5 1 . Riley, R. L., Bacteriol. Rev., 25, 24348 (1961)
152. Smith, C. E., Pappagianis, D., Levine,
H. B., Saito, M., Bacteriol. Rev.,
25, 3 10-20 (1961)
1 53. Converse, J. L., Reed, R. E., Bacteriol,
Rev., 30, 678-94 (1966)
154. Sas!aw, S., Carlisle, H. N., Sparks, J.,
Proc. Soc. Exptl. Bioi., Med., 103,
342-44 (1960)
155. Furculow, M. L., Bacteriol. :Rev., 25,
301-9 (1961)
156. Davenport, F. M., Bacteriol. Rev., 25,
294-300 (1961)
1 5 7. Schulman, J. L., Kilbourn!', E. D.,


1 59.

1 60.
1 62.
1 64.






A erobiology,

141-45, Berkeley, Calij., 1263. (The

Naval BioI. Lab., Univ. of Calif.,
405 pp., 1963)
Saslaw, S., Proc. Intern. Symp. A ero
biology, 1st, 147-50, Berkeley, Calif.,
1963. (The Naval BioI. Lab., Univ.
of Calif., 405 pp., 1963)
M ill er, W. S., Demchak, P., Rosen
berger, C. R., Dominik, J. W.,
Bradshaw, J. L., Am. J. Hyg., 77,
1 14-21 (1963)
Saslaw, S., Carlisle, H. N., Bacteriol.,
Rev., 30, 636-44 (1966)
Lamanna, C., Science, 130, 763-72
Lamanna, C., Bacteriol. Rev., 2 5, 32330 (1961)
Imshenetsky, A. A., Bull. A tomic Sci.,
16, 241-42 (1960)
Lwoff, A., BUll. A tomic Sci., 16, 24344 (1960)
Overholt, E. L., Tigertt, W. D.,
Kadull, P. J., Ward, M. K., A m . J.
Med., 30, 785-806 (1961)
Plotkin, S. A. , Brachman, P. S., Utell,
M., Bumford, F. H., Atchison, M.
M., Am. J. Med., 29, 992-1001
Meyer, K. F., Pasteurella and Fran
cisella, in Bacterial and Mycotic In
jections of Man. (Dubos, R. J.,
Hirsch, J. G., Eds., Pitman Med.
Publ. Co., Ltd., London, 1025 pp.,
McCrumb, F. R., Mercier, S., Robic,
J., Bonillat, M., Smadel, J. E.,


Woodward, R. T., Goodner, K.,

Am. J. Med., 14, 284-93 (1953)
1 69. McCrumb, F. R., Bacteriol. Rev., 25,

283 (1961)
1 70. Crozier, D., Tigertt, W. D., Cooch,
J. W., Reference 8 of Ann. Rept.,
1958, of Sect. II, U.S. Army Med.
Unit, Ft. Detrick, Md., in Symp. on
Military Med. Am. Med. Assoc.
Meeting, June IS, 1960 (see also
Ref. 239)
1 7 1 . Fremont-Smith, P., Antibiotics Ann.
1958-1IJ59, 840-42 (1959)
1 72. Gasper, A. J., Tressell, H. B., Ward,
M. K., J. Bacteriol., 82, 564-69
1 73. Ward, M. K., Military Med., 128,
1 00-1 (1963)
1 74. Heden, C.-G., Memorandum on auto
mation of microbiological tech
niques. Conf. Rapid Detection
Methods for Airborne Bacteria and
Viruses, Stockholm, 1966. Intern.
lnst. Peace and Conflict Res.,
(SIPRI), Stockholm, 1966)
1 75. Gillespie, D., Spiegelman, S., J. Mol.
Bioi., 12, 829-42 (1965)
1 76. Ritter, D. B., Gerloff, R. K., J. Bact
teriol. , 92, 1 838-39 (1966)
1 77. Mansberg, H. P., Kusnetz, J., J.
Histochem. Cytochem., 14, 260-73
1 78. Jaeger, R. F., Spertzel, R. 0., Kuehne,
R. W., APPI. Microbiol. , 9, 585-87
1 79. Dozier, S. M., Military Med., 128,
97-99 (1963)
180. McKinney, R. W., Military Med. , 128,
102-3 (1963)
181. Cherry, W. B., Goldman, M., Carski,
T. R., Moody, M. D., Fluorescent
Antibody Techniques in the Diag
nosis of Communicable Diseases.
U.S. Public Health Servo Publ. 729
182. Diagnostic Procedures and Reagents,
3rd Ed. (Am. Public Health Assoc.,
New York, 589 pp., 1950)
183. Manual on Microbiological Methods ;
Committee on Bacteriological Tech
niques of Am. Soc. Microbiologists.
(McGraw Hill, New York, 1957)
184. Lennette, E . H., in Diagnostic Pro
cedures for Virus and Rickettsial
Diseases, 2nd ed., 1-52. (Francis,
T., Jr., Ed., Am. Public Health

Assoc., 578 pp., New York 1956)

185. Lennette, E. H., Kaiser Found. M.d.
Bull., 7, 12-24 (1959)
186. Cherry, G. B., Kemp, S. D., Parker,
A., Prog. Ind. Microbial., 4, 37-60



187. Guyton, H. G., Lense, F. T., A rch. Ind.

Health, 14, 246-49 (1956)
188. Guyton, H. G., Decker, H. M., Anton,
G. T., Arch. Ind. Health, 20, 91-95
189. Guyton, H. G., Buchanan, L. M.,

Lense, F.


A ppl. Microbiol. , 4,

141-43 (1956)
190. Derr, J. S., Jr., Proc. Intern. Symp.
A erobiology, 1st 227-61, Berkeley,

Annu. Rev. Microbiol. 1967.21:639-676. Downloaded from

Access provided by Universidade Federal de Alagoas on 01/02/16. For personal use only.


1 963. (The Naval BioI. Lab.,

Univ. of Calif., 405, pp. 1963)

191. Zimmerman, L., Proc. Intern. Symp.
A erobiology, 1st 285-87, Berkeley,
Calif. , 1965. (The Naval BioI. Lab.,
Univ. of Calif., 405, pp. 1963)
192. Cox, C. S., Proc. Intern. Symp. A ero
biology, 1st, 345-63, Berkeley, Calif.,

1f)63. (The Naval BioI. Lab., Univ.

of Cal i f. , 405, pp. 1963)
193. Hatch, M. T., Dimmick, R. L., Proc.
Intern. Symp. A erobiology, 1st, 26577, Berkeley, Calif., 1f)63. (The
Naval Bioi. Lab., Univ. of Calif.,
405 pp., 1963)
194. Hatch, M. T., Dimmick, R. L., Bac
teriol. Rev., 30, 597-602 (1966)
195. Webb, S. J., Can. J. Microbiol., 6,
7 1-87 (1960)
196. Webb, S. J., Can. J. Microbiol., 6,
89-105 (1960)
197. Hess, G. E., A PPI. Microbiol. , 13, 78187 (1966)
198. Glassman, H., Bacteriol. Rev., 25, 36061 (1961)
199. Stakman, E. C., Am. Scientist, 35,
32 1-50 (1948)
200. Langmuir, A. D., Bacteriol.. Rev., 25,
1 73-81 (1961)
201. Gorelick, A. N., Bacteriol. Rev., 30,
644--45 (1966)
202. Buckland, F. E., Bynoe, M. E., Tyrell,
D. A. J., J. Hyg., 63, 327-43 (1965)
203. Harper, G. L., Proc. Intern. Symp.
A erobiology 1st, 335-41, Berkeley,
Calif. , If)63. (The Naval Bio!. Lab.,
Univ. of Calif., 405 pp., 1963)
204. Jensen , M. M . , Proc. Intern. Symp.
A erobiology, 1st, 2 19-24, Berkeley,
Calif., 1963. (The Naval BioI. Lab.,
Univ. of Calif., 405 pp., 1963)
205. Webb, S . J., Proc. Intern. Symp. A ero
biology, 1st, 369-77, Berkeley, Calif. ,

1M3. (The Naval Bio!. Lab., Univ.

of Calif. , 405 pp., 1963)
206. Watkins, H. M. S., Gold berg, L. J.,
Deig, E. F., Leif, W. R., Proc.

Intern. Symp. A erobiology, 1st, 38188, Berkeley, Calif., Oct 2-5, 1 963.
(The Naval BioI. Lab., Univ. of
Calif., 405 pp., 1963)

207. Gochenour, W. S., Jr., Military Med. ,

1 2 8, 86-89 (1963)
208. Riehl, H., Bull. A tomic Sci., 16, 253
209. Perkins, W. A., Vaughan, L. M., Bac
teriol. Rev. , 25, 347-55 (1961)
210. Silver, 1. H., Proc. Intern. Symp. A ero
biology, 1st, 319-26, Berkeley, Calif.,
If)63. (The Naval Bio!. Lab., Univ.
of Cal if. , 405 pp., 1963)
2 1 1 . Lion, M. B., J. Gen. Microbiol., 32,
321-29 (1963)
2 12. Zimmermann, L., J. Bacteriol. , 84,
1297-1302 (1962)
2 1 3. Won, W. D., Ross, H. A ppl. Micro
bioi., 14, 742-45 (1966)
214. Zentner, R. J., Bacteriol. Rev., 30,
551-57 (1966)
215. Maltman, J. R., Proc. Intern. Symp.
A erobiology, 1st. 291-301, Berkeley,
Calif., 1M3. (The Naval BioI. Lab.,

Univ. of Calif., 405 pp. 1963)

2 1 6. Glassman, H . , Bacteriol. Rev., 25,
362-77 (1961)
2 1 7 . Phillips, C. R., A n tiseptics, Disinfec
tants, Fungicides and Sterilization,
2nd ed., 746-65 (Reddish, G. F.,

E d ., Lea & Febiger,

2 1 8. Hoffman , R. K., Warshowsky, B.,
Appl. Microbiol., 6, 358-62 (1958)
219. Edwards, R. W., Favero, M . S., Hoff
man, R. K., Lanahan, T. B . , Mac
Leod, N. H., McDade, J. J., Skaliy,
P., Briggs Phillips, G., A biblio
graphy on 'lJapor phase disinfectants.

(Am. Assoc. for Cont. Control,

10 pp., M arch , 1965)
220. Schley, D. G., Hoffman, R. K., Phil
lips, C. R., A ppl. Microbiol., 8, 1519 (1960)
2 2 1 . Spiner, D. R., Hoffman, R. K., Appl.
Microbiol., 8, 152-55 (1960)
222. Decontamination after biological war
fare attack. Appendix NP 24-31 to
Nat!. BioI. and Chern. Defense
Plan. (Office of Civil and Defense
Mobilization, Washington, D.C.,

223. Levin, M. Ye., Malinin, G. A., Man
drazhitskiy, M. M., Sinitsyn, V. P.,
Fedorov, V. I . , Defense A gainst
A gents of Mass Destruction. (State
Educational and Pedagogic Pub!.

House, Ministry of Educ., Mos

cow, 1958. Trans . by U.S. Joint
Pub!. Res. Serv., New York)

224. Baratov, G. F., Local Anti-A ir De

fense of the Population under con
ditions of Chemical, A tomic and
Biological A ttack. (State Med. Pub!.


House USSR, Kiev, 1959. Trans.
by U.S. Joint Pub!. Res. Serv., New
225. Haas, V. H., J. Am. Med. Assoc., 145,

Armees Terre, Mer, Air,


Annu. Rev. Microbiol. 1967.21:639-676. Downloaded from

Access provided by Universidade Federal de Alagoas on 01/02/16. For personal use only.

Eveland, W. C., Vorder Bruegge,

C., Smetana , H. F., Trans. N. Y.
Acad. Sci., 22, 323-33 (1960)

235. Sawyer, W. D., Dangerfield, H. G.,

Hogge, A. L., Crozier, D., Bacteriol.
Rev., 30, 542-48 (1966)
236. Ley, H. L., Jr. , Diercks, F. H" Pater

son, P. Y., Smadel, J. E., Wisse

man, C. L., Traub, R., Am. J. Hyg.,
56, 303-12 (1952)
237. Ley, H. L., Jr., Smade1, J. E., A nti





238. Wedum, A. G., J. Am. Med. A ssoc.,

162, 34-37 (1956)
239. Crozier, D., Tigertt, W. D., Cooch,

J. W. Symp. on medical importance

of chemical, biological and radio
logical warfare, Section on Military

Medicine. (Am. Med. Assoc. meet

ing, June 15, 1960)
240. Bawden, F. C., Bull. A tomic Sci., 1 6,

247-49 (1950)
241. Averbuck, V. L., Berlin, I. Z., Protec
tion of Grain Products from Radio
active, Chemical and Bacterial Sub
stances. (State Educational and

Pedagogic Pub!. House, Ministry

of Educ., Moscow, 1963. Trans. by
U.S. Joint Pub!. Res. Serv., Wash
ington, D.C.)
242. Keiser, R. A., Univ Penn. Bull., Vet.

Extension Quart., 1 15, 50-59 (1949)

243. Todd, F. A., The Military Surgeon,
1 12, No. 3 (1953)
244. Isachenko, V. M., Rev. Int. Servo Sante

39, 159-60

245. Kovalenko, V. Ya., Protecting Food

and Water A gainst Chemical, Bio
logical and Radiological A ttack.

900-5 (1951)

226. Disaster preparedness for pharmacists.

J. Am. Pharm. Assoc., 6, 80-86
227. Douglas, R. F., Military Med., 128,
104-9 (1963)
228. Overholt, E . L., Tigertt, W. D., Radio
logy, 74, 758-65 (1960)
229. Munter, E. J., J. Am. Med Assoc.,
128, 281-83 (1945)
230. Burmeister, R. W., Tigertt, W. D.,
Overholt, E. L., Ann. Internal
Med., 56, 789-800 (1962)
231. Jacobson, G., Delinger, R. B., Carter,
R. A., Radiology, 53, 739-48 (1949)
232. Blount, R. E., Military Med., 128,
129-3 1 (1963)
233. Raska, K., World Health Organ.
Chronicle, 20, 3 1 5-21 (1966)
234. Tigertt, W. D.,
Berge, T. 0.,
Gochenour, W. S., Gleiser, C. A.,


(Medgiz, Moscow, 1963. Trans. by

U.S. Joint Publ. Res. Serv., Wash
ington, D.C.)
246. Lamoureux, V. C., Southwest Water
works J., 33, 4-5 (1951)
247. Mutschin, A., Zivilschutz, 12, 506-13
248. Fothergill, L. D., Military Med., 128,
132-34 (1963)
249. Langmuir, A. D., New Engl. J. Med.,
2 68, 1 82-92 (1963)
250. Raska,
K., Z. Hyg. Epidemiol.
(Prague), 8, 137-68 (1964)
2 5 1 . Shih-Shan Fang, Chinese Med. J., 70,
329-32 (1952)
252. Raska, K., Kaplan, M., Disarmament

in microbiological warfare for small

powers through international guar
antees of assistance in case of at
tack, Proc. Pugwash Conf. Sci.,
World Affairs, 14th, 305-7, Venice,

Italy, April, 1965

253. Kaplan, M., Inspection for allegations

of use of microbiological weapons,

Proc. Pugwash Conf. Sci., World
Affairs, 14th, 284-88, Venice, Italy,
April, 1965

Heden, C.-G., Leerhfllj , M., Malek, 1.,

M ose, O., Report on experimental
non-production control of biological
weapons to Standing Committee of
the Pugwash Organization, Sept.
1, 1966
255. Holst, H. E. von, Liikartidningen, 63,

4733-39 (1966)
256. Zetterberg, B., in Forsvars- och Ka
tastrofmedicin, 155-66. (Medicin

alstyrelsens Sjukvardsberedskaps
namnd, Stockholm, 1966)
257. Notification No. 1 16 : Medicinsk KaPubl.
Health, Stockholm, 1966)
258. Knorr, K. Supranational versus in
ternational models for general and
complete disarmament, in The
Strategy of World Order. (Falk, R.
A., Mendlowitz, S. H., Eda., World
Law Fund)
259. Lederberg, J.. The Washington Post,
Sept. 24 (1966)
260. West, M., Children of the Sun. (Heine
mann, Ltd., London, 188 pp., 1957)
261. Lamanna, C., Am. J. Public Health,
243-53 (1966)
262. Butlin, K., New Scientist, 14, 804-6

Annu. Rev. Microbiol. 1967.21:639-676. Downloaded from

Access provided by Universidade Federal de Alagoas on 01/02/16. For personal use only.



263. Starr, M . P., Ed., Proc. Conference

Global Impacts of Applied Micro
biology, Stockholm, 1963. (Almq
vist & Wicksell, Stockholm, J.
Wiley & Sons, Inc., New York, 572
pp., 1964)
264. Heden, C.-G., Starr, M. P., . Advan.
A ppl. Microbiol., 6, 1-26 (1964)
265. Meselson, M . , A proposal to inhibit
the development of biological weap
ons, Proe. Pugwash Coni. Sci.,
World Affairs, 14th, 29 7-304, Ven
ice, Italy, April, 1965
266. Groupe, V., On the feasibility of con
trol of biological warfare, in In
spection lor Disarmament, 185-90.
(Melman, S., Ed., Columbia Univ.
Press, New York , 291 pp., 1958)
267. Proceedings of Pugwash Conference
of International Sci entists on Bi
ological and Chemical Warfare,
Pugwash, Nova Scotia, Canada,
August 24-30, 1959
268. M arcovi ch , II., Control of biological
weapons. Proe. Pugwash Conf. Sci.,
World Affairs, 14th, 293-96, Ven
ice, Italy, April, 19155

269. Heden, C.-G., The possibility of usi ng

a mechanized documentation cen
ter to provide data on the basis of
which inspections can be planned,
Proe. Pugwash Coni. Sci., World Af

fairs, 14th, 274-84, Venice, Italy,

April, 1265

270. Kaplan, M., Definition of defensive as

opposed to offensive work on mi
crobiological weapons, Proc. Pug
wash Con!. Sci., Warld Affairs. 14th,
282-83, Venice, Italy, April, 1965
2 7 1 . Maaj1lle, 0., On th e problems of dec1as-

sification of BW research, Proe.

Pugwash Conf. Sci., World Affairs,
14th, 289-92, Venice, Italy, April,

272. Heden, C.-G., The use of existing

channels for guarding against mili
tary applications of newly acquired
microbiological knowledge, Proc.
Pugwash Con!. Sci. , World Affairs,
14th, 258-73, Venice, Italy, April,

273. Goldwasser, R., M icrobiological iden

tification techniques in investiga
tion of alleged use of microbiological
weapons, Proc. Pugwash Conf. Sci.,
World Affairs, 14th, 255-57, Ven
ice, Italy, A pril, 1965
274. Problems of Biological warfare, Pug
wash Newsletter, 4, No. 1 82, 9 (1966)
275. Bjornerstedt, R., Konkret, 1 , 45-46, 8 1
276. Meselson, M. Detection of remote
biological weapons tests-a prelimin
ary assessment. Working document

to The I ntern. lnst. for Peace and

Conflict Res. (SlPRl), Stockh olm,
Dec. 1966

277. Thorne, D. S., Proe. Intern. SymP .

Aerobiology, 1st, 1 33-39, Berkeley,
Calif., Oct. 1963 (The Naval BioI.
Lab., Univ. of Calif., 405 pp. 1963)
278. Cabelli , V. J., Hodapp, F. A., Fergu
son, E. W., McElmury, R. C.,
Prot. Intern. Symp. A erobiology, 1st,
1 5 1-79, Berkeley, Calif., Oct. 1963

(The Naval BioI . Lab., Univ. of

pp., 1963
279. Szilard, L., The Voice of the Dolphins.
(Simon & Schuster, New York, 122
pp., 1961)
Calif., 405