ArticlePDF Available
BACTEEzOLOGICAL
REviEws,
Dec.
1976,
p.
793-802
Copyright
0
1976
American
Society
for
Microbiology
Vol.
40,
No.
4
Printed
in
U.S.A.
"Who
Discovered
Bacteriophage?"
DONNA
H.
DUCKWORTH
Department
of
Immunology
and
Medical
Microbiology,
University
of
Florida,
Gainesville,
Florida
INTRODUCTION..............................................................
793
TWORrS
DISCOVERY
.........
............................
794
D'HERELLE'S
DISCOVERY
...............
......................
795
THE
DISCOVERY
OF
TWORTS
DISCOVERY
...............................
797
THE
CONTROVERSY
GRIOWS
..................
...................
798
D'HERELLE
IS
DISCREDITED
.
.....................................
799
DID
D'HERELLE
DISCOVER
PHAGE
IN
1910?
..............................
800
CONCLUSIONS
...........
................................
800
LITERATURE
CITED
.....................................
801
INTRODUCTION
Although
most
authors
state
that
Twort
in
1915
and
d'Herelle
in
1917
independently
dis-
covered
bacterial
viruses
(8,
9,
26),
some
have
made
allusions
which
suggest
that
d'Herelle
may
not
have
been
altogether
honest
in
claim-
ing
to
have
had
no
knowledge
of
Twort's
1915
discovery
when
he
published
his
1917
work.
In
his
excellent
book,
The
Molecular
Biology
of
Bacterial
Viruses,
Gunther
Stent
writes:
"Gra-
tia
[sic]
drew
attention
to
Twort's
forgotten-
or,
rather,
never
noted"
-
discovery
(33).
And
in
his
introduction
to
Phage
and
the
Origins
of
Molecular
Biology,
a
similar
ambiguity
ap-
pears:
"Bacterial
viruses
were
discovered
in
1915
by
the
English
microbiologist
F.
W.
Twort,
and
two
years
later-perhaps
independently,
perhaps
not-by
the
French-Canadian
F.
d'Herelle"
(34).
Stent
had
no
proof
that
d'Herelle
had
been
dishonest
(G.
Stent,
per-
sonal
communication),
but
was
very
much
in-
fluenced
by
Andre
Lwoff
who
felt
that
d'Herelle
may
have
been
dishonest
(G.
Stent,
personal
communication;
A.
Lwoff,
personal
communi-
cation).
Since
I
thought
the
idea
that
d'Herelle
had
had
knowledge
of
Twort's
work
was
com-
pletely
incompatible
with
the
joyous
enthusi-
asm
with
which
he
described
his
discovery
(18),
I
investigated
the
matter
to
see
how
this
accu-
sation
might
have
arisen
and
to
see
whether,
indeed,
I
could
discover
who
discovered
bacte-
riophage.
It
would
seem,
on
the
surface,
that
who
discovered
what
would
be
a
relatively
sim-
ple
matter
to
resolve.
Actually,
though,
the
pages
of
the
history
of
science
are
strewn
with
disputes,
often
sordid,
over
priority
(31).
It
ap-
pears
that
the
territorial
imperative
is
as
real
in
the
Kingdom
of
Science
as
it
is
in
the
jungles
of
Africa.
To
unravel
the
many
possible
reasons
for
these
"territorial"
disputes
is
rather
more
diffi-
cult
than
to
prove
that
they
exist.
Their
origins
undoubtedly
lie
in
a
complex
of
unresolved
questions
involving
the
sociology
and
philoso-
phy
of
science.
The
disputes
may,
for
instance,
arise
from
the
actual
frequency
of
simultaneous
discovery.
But
to
understand
why
so
many
dis-
coveries
are
made
by
different
individuals
in-
dependently
at
the
same
time
depends
on
an
in-
tricate
philosophical
analysis
of
the
way
in
which
science
progresses.
Discoveries
may
not
be
simple
events
occurring
at
definite
time
points,
but
rather
may
result
from
more
elabo-
rate
extended
processes
involving
large
num-
bers
of
people.
This
has
been
extensively
dis-
cussed
by
Thomas
Kuhn
(28).
In
this
view,
claims
to
priority
would
be
somewhat
artificial,
if
not
meaningless.
One
could
also
invoke
egotism
as
the
cause
of
the
many
acrimonious
controversies
over
prior-
ity
that
scientists
have
engaged
in.
The
sociolo-
gist,
Robert
Merton,
has
rejected
this
idea,
however,
and
sees
the
disputes
as
"signposts
announcing
the
violation
of
the
social
norms"
of
the
scientific
establishment.
In
his
view,
origi-
nality
in
science
is
at
a
premium,
and
rewards
come
from
demonstrations
of
originality.
But
humility
and
allocation
of
credit
is
also
an
insti-
tutional
norm.
Under
the
stresses
imposed
by
the
system,
the
balance
between
these
opposing
values
may
be
upset,
and
pathogenic
concern
with
original
discovery
may
result
in
"conten-
tiousness,
self-assertive
claims,
secretiveness
."
(31).
Many
disputes
also
surely
arise
from
the
lack
of
a
clear-cut
definition
of
what
constitutes
dis-
covery.
Is,
for
instance,
an
observation
a
discov-
ery,
or
must
all
the
ramifications
of
the
obser-
vations
be
recognized
to
constitute
a
discovery?
These
questions,
however
-
what
is
discovery
and
what
is
the
origin
of
the
fervor
to
be
first-
are
beyond
the
scope
of
this
paper.
As
it
turns
out,
even
the
question
posed
in
the
title
is
not
answered.
Instead,
it
is
my
intent
to
present
793
794
DUCKWORTH
some
of
the
heretofore
neglected
circumstances
surrounding
the
discovery
of
bacteriophage
and,
I
hope,
to
clear
the
name
of
Felix
d'Herelle
from
the
aspersions
that
have
been
cast
upon
it.
(This
paper
is
part
of
an
address
entitled
"The
Filterable
History
of
Discovery,"
pre-
sented
at
the
University
of
Florida
on
2
March
1976
as
one
of
the
President's
Scholars
Lecture
Series.)
TWORT'S
DISCOVERY
F.
W.
Twort,
the
son
of
a
country
doctor
and
a
hard-working
mother,
was
trained
in
medicine
at
St.
Thomas
Hospital
in
London,
but
as
the
fortunes
of
his
family
were
small
he
was
unable
to
pursue
his
keen
interest
in
pathology.
"It
was
imperative,"
relates
Twort
in
his
article,
"The
Discovery
of
the
Bacteriophage"
(37),
"that
I
should
earn
at
least
sufficient
to
be
able
to
pay
for
lodgings
and
food.
Accordingly,
I
accepted
the
first
paid
post
available,
which
was
that
of
Assistant
Superintendent
of
the
clinical
laboratory
at
the
hospital."
The
post
enabled
him,
nonetheless,
to
gain
experience
in
pathology,
and
in
1909
Twort
was
appointed
Superintendent
of
the
Brown
Institution,
Lon-
don
University.
The
Brown
Institution
had
been
founded
in
1871
to
provide
a
hospital
"for
the
care
and
treatment
of
Quadrupeds
or
Birds
useful
to
man."
In
his
capacity
as
direc-
tor,
Twort
was
allowed
to
carry
out
research
in
any
branch
of
pathology
or
bacteriology,
"pro-
vided
expenditure
could
be
kept
within
the
lim-
its
of
the
small
income
available."
He
chose
to
study
the
growth
requirements
of
primitive
forms
of
life.
He
had
one
success,
obtaining
an
essential
substance
that
would
allow
Johne's
bacillus,
the
causative
agent
of
a
serious
dis-
ease
of
cattle,
to
grow
in
artificial
medium.
His
essential
substance
has
since
been
identified
as
vitamin
K
(20,
37).
He
then
went
on
to
perform
experiments
in-
tended
to
indicate
how
viruses
could
be
grown
in
artificial
media.
This
seems
an
unfortunate
avenue
of
research
to
us
as
we
now
know
that
viruses
can
only
grow,
if
indeed
we
can
say
they
grow
at
all,
inside
living
cells.
But
Twort
was
proceeding
upon
a
not
unreasonable
hypothesis
which
was
that,
since
viruses
were
the
small-
est,
hence
simplest,
forms
of
life,
they
must
have,
at
one time
in
evolutionary
history,
been
able
to
grow
in
a
medium
devoid
of
any
living
matter.
As
Twort
explains
in
his
famous
note
to
the
Lancet
on
4
December
1915
(35),
"attempts
to
cultivate
these
(viruses)
from
such
materials
as
soil,
dung,
grass,
hay,
and
water
from
ponds
were
made
on
specially
prepared
media.
It
is
impossible
to
describe
all
these
in
detail
but
generally
agar,
egg,
or
serum
was
used
as
a
basis,
and
to
these
varying
quantities
of
certain
chemicals
or
extracts
of
fungi,
seeds,
and
etc.,
were
added."
He
would
then
inoculate
these
various
media
with
extracts
of
his
various
soils,
dung,
or
sera,
which
had
been
filtered
through
candle
filters
to
remove
all
bacteria.
Every
one
of
his
hundreds
of
experiments
was
negative.
He
never
succeeded
in
growing
a
filter-passing
agent
on
artificial
media.
He
did,
however,
ob-
serve
something
else.
He
had
inoculated
an
agar
medium
with
some
fluid
(unfiltered)
com-
monly
used
for
smallpox
vaccinations.
He
no-
ticed
that
although
the
vaccinia
virus
did
not
grow,
a
bacteria
-
a
micrococcus
-
did
grow.
The
bacteria,
however,
appeared
to
be
afflicted
with
some
disease
-"inoculated
agar
tubes
of-
ten
showed
watery-looking
areas,
and
in
cul-
tures
that
grew
micrococci
it
was
found
that
some
of
these
colonies
could
not
be
subcultured,
but
if
kept
they
became
glassy
and
transpar-
ent."
In
retrospect,
this
phenomenon,
which
came
to
be
known
as
the
glassy
transformation
of
Twort,
seems
exceedingly
strange.
It
is
so
rare
that,
in
the
more
than
14
years
that
I
have
worked
with
bacteria
and
their
viruses,
I
have
never
observed
it-
at
least
that
I
know
of.
Fur-
thermore,
I
have
seen
only
one
reference
in
the
literature
to
a
similar
phenomenon,
and
that
was
Sir
Alexander
Fleming's
observation
that
what
was
later
found
to
be
penicillin
caused
colonies
of
staphylococcus
to
appear
like
"drops
of
dew"
(2).
Twort,
however,
made
some
very
interesting
observations
about
the
glassy
trans-
formation.
(i)
The
affected
colonies
would
not
grow
on
any
medium.
(ii)
Examination
of
the
glassy
areas
revealed
only
minute
granules
and
no
bacteria.
(iii)
If
a
pure
culture
of
mi-
crococcus
was
touched
with
a
small
portion
of
one
of
the
glassy
colonies,
the
growth
at
the
point
touched
started
to
become
transparent
and
gradually
made
the
whole
colony
trans-
parent.
(This
is
the
startling
part
of
his
obser-
vation as
regards
bacterial
viruses.
Usually
by
the
time
bacteria
have
grown
long
enough
to
form
a
visible
colony,
they
have
reached
a
phase
such
that
they
cannot
be
attacked
by
a
virus.)
(iv)
After
filtration
of
the
glassy
mate-
rial
through
a
Chamberland
candle,
it
retained
its
ability
to
cause
the
"glassy
transformation."
(v)
The
"transformation"
could
be
conveyed
to
fresh
cultures
for
an
indefinite
number
of
gen-
erations.
Twort
concluded
that
the
cause
of
the
glassy
transformation
was
an
infectious,
filterable
agent
that
killed
bacteria
and
in
the
process
multiplied
itself.
He
could
have
very
logically
BACTERIOL.
REV.
WHO
DISCOVERED
BACTERIOPHAGE?
795
concluded
that
he
was
dealing
with
a
bacterial
virus.
But
he
does
not
do
so.
He
says:
from
these
results
it
is
difficult
to
draw
definite
conclusions....
[This]
may
be
living
proto-
plasm
that
forms
no
definite
individuals
or
an
enzyme
with
the
power
of
growth....
In
any
case,
whatever
explanation
is
accepted,
the
pos-
sibility
of
its
being
an
ultra-microscopic
virus
has
not
been
definitely
disaproved
because
we
do
not
know
for
certain
the
nature
of
such
a
virus.
Then
a
little
later,
"On
the
whole
it
seems
probable,
though
by
no
means
certain,
that
the
active
transparent
material
is
produced
by
the
micrococcus,
and
since
it
leads
to
its
own
de-
struction
and
can
be
transmitted
to
fresh
healthy
cultures
it
might
almost
be
considered
as
an
acute
infectious
disease
of
micrococci."
At
the
end
of
the
paper
he
laments,
"I
regret
that
financial
considerations
have
prevented
my
carrying
these
researches
to
a
definite
conclu-
sion"
(35).
Here,
as
happened
also
with
the
discovery
of
plant
viruses
and
animal
viruses,
we
have
a
remarkable
discovery
faced
by
its
discoverer
with
uncertainty.
Although
it
was
by
this
time
known
that
there
were
filterable
viruses
that
could
grow
in
either
plant
cells
or
animal
cells,
Twort
was
reluctant
to
believe
he
had
discov-
ered
a
bacterial
virus-if
indeed
he
had-al-
though
he
did
consider
this
a
possibility.
Hence
he
considered
Beijerinck's
old
idea
of
a
fluid
form
of
life,
introduced
to
explain
the
filterabil-
ity
of
the
agent
of
tobacco
mosaic
disease
(3),
and
also
the
possibility
that
the
micrococcus
was
producing
an
enzyme
that
could
grow
and
multiply.
This
latter
idea,
which
seems
ridicu-
lous
now,
was
the
focal
point
of
a
controversy
that
raged
in
the
scientific
literature
for
many
years.
The
controversy
did
not
start
immedi-
ately,
however.
In
fact,
Twort's
paper
went
un-
recognized
until
26
March
1921-5
years,
3
months,
and
22
days.
D'HERELLE'S
DISCOVERY
Felix
d'Herelle,
the
man
who
was
heralded
for
4
years
as
the
sole
discoverer
of
bacterial
viruses-or
bacteriophage,
as
he
called
them
-
lived
a
life
that
was
more
befitting
of
a
Magel-
lan,
a
true
explorer,
than
a
mere
bacteriologist.
Born
in
1873
in
Montreal
of
a
French-Canadian
father
and
a
Dutch
mother,
he
was
educated
in
France
and
Holland,
and
during
his
life
trav-
eled
and
worked
in
Guatemala,
Mexico,
South
America,
Egypt,
Algeria,
Tunisia,
Indochina,
and
Russia.
He
was
a
professor
at
Yale
for
5
years,
and
an
associate
of
his
there
has
recently
said
of
him
that
everywhere
d'Herelle
was,
there
were
fireworks
(Florence
Mack,
personal
communication).
The
fireworks
materialized
rather
unfortunately
in
Russia,
where
d'Herelle
went
several
times
during
the
1930s
to
found
institutes
for
the
study
of
bacteriophage.
Dur-
ing
one
of
his
trips
his
trusted
associate,
Eliava,
was
arrested
and
shot
(1,
7,
29,
32).
He
left
Yale
for
reasons
that
are
not
entirely
clear,
and
"several
confidential
letters
exchanged
be-
tween
Dean
Winternitz
and
Professor
d'Herelle
cannot
be
released"
(A.
Ebbert,
personal
com-
munication).
It
was
in
Mexico
that
d'Herelle
first
observed
what
later
came
to
be
called
bacteriophage.
His
account
of
this
discovery
written
in
1949-
about
40
years
after
he
had
made
the
discovery
-
is
rarely
equalled
in
the
scientific
literature
of
today
for
its
ingenuous
enthusiasm
(18).
He
said:
In
1910,
I
was
in
Mexico,
in
the
state
of
Yuca-
tan,
when
an
invasion
of
locusts
occurred;
the
Indians
reported
to
me
that
in
a
certain
place
the
ground
was
strewn
with
the
corpses
of
these
insects.
I
went
there
and
collected
sick
locusts,
easily
picked
out
since
their
principal
symptom
was
an
abundant
blackish
diarrhoea.
This
malady
had
not
as
yet
been
described,
so
I
stud-
ied
it.
It
was
caused
by
bacteria,
the
locust
coccobacilli,
which
were
present
almost
in
the
pure
state
in
the
diarrhoeal
liquid.
I
could
start
epidemics
in
columns
of
healthy
insects
by
dust-
ing
cultures
of
the
coccobacillus
on
plants
in
front
of
the
advancing
columns:
the
insects
in-
fected
themselves
as
they
devoured
the
soiled
plants.
During
the
years
which
followed,
I
went
from
the
Argentine
to
North
Africa
to
spread
this
illness.
In
the
course
of
these
researches,
at
various
times
I
noticed
an
anomaly
shown
by
some
cultures
of
the
coccobacillus
which
in-
trigued
me
greatly,
although
in
fact
the
obser-
vation
was
ordinary
enough,
so
banal
indeed
that
many
bacteriologists
had
certainly
made
it
before
on
a
variety
of
cultures.
The
anomaly
consisted
of
clear
spots,
quite
circular,
two
or
three
millimetres
in
diameter,
speckling
the
cultures
grown
on
agar.
I
scratched
the
surface
of
the
agar
in
these
trans-
parent
patches,
and
made
slides
for
the
micro-
scope;
there
was
nothing
to
be
seen.
I
concluded
from
this
and
other
experiments
that
the
some-
thing
which
caused
the
formation
of
the
clear
spots
must
be
so
small
as
to
be
filtrable,
that
is
to
say,
able
to
pass
a
porcelain
filter
of
the
Chamberland
type,
which
will
hold
back
all
bacteria.
However,
the
appearance
of
these
clear
patches
was
inconstant.
I
sometimes
went
weeks
without
seeing
a
single
one,
and
I
could
not
reproduce
the
phenomenon
at
will;
I
there-
fore
could
not
study
it.
VOL.
40,
1976
796
DUCKWORTH
In
March,
1915,
during
the
first
World
War,
a
large
invasion
of
locusts
appeared
in
Tunisia,
threatening
to
destroy
the
harvests
which
were
then
so
vital;
I
was
given
the
job
of
starting
an
epidemic
amongst
them.
As
the
result
of
the
infection
there
was
a
considerable
mortality,
and,
even
more
interesting,
when
the
following
year
all
the
rest
of
North
Africa
was
again
invaded,
Tunisia
remained
free.
In
the
course
of
this
campaign,
I
again
ob-
served
my
clear
spots,
and
before
returning
to
France
I
stayed
for
a
time
at
the
Institut
Pas-
teur
in
Tunis,
to
investigate
their
significance.
I
showed
them
to
Charles
Nicolle,
then
direc-
tor
of
the
Institute,
and
he
said
to
me:
'That
may
be
the
sign
of
a
filtrable
virus
carried
by
your
coccobacilli,
a
filtrable
virus
which
is
the
true
pathogenic
agent,
while
the
coccobacillus
is
only
a
contaminant.'
So
I
filtered
emulsions
of
cultures
grown
on
agar
and
showing
the
clear
spots,
and
tried
to
infect
healthy
locusts
with
the
filtrate,
but
without
result.
On
my
return
to
Paris
in
August
1915,
I
was
asked
by
Dr.
Roux
to
investigate
an
epidemic
of
dysentery
which
was
raging
in
a
cavalary
squadron,
then
resting
at
Maisons-Laffitte.
I
thought
the
hypothesis,
put
forward
for
the
lo-
custs'
illness
might
be
helpful
in
understanding
human
dysentery.
I
therefore
filtered
emulsions
of
the
faeces
of
the
sick
men,
let
the
filtrates
act
on
cultures
of
dysentery
bacilli
and
spread
them
after
incubation
on
nutritive
agar
in
petri
dishes:
on
various
occasions
I
again
found
my
clear
spots,
but
the
feeding
of
these
cultures
to
guinea
pigs
and
rabbits
produced
no
disease.
At
this
time
we
often
got
cases
of
bacillary
dysentery
in
the
hospital
of
the
Institut
Pasteur
in
Paris.
I
resolved
to
follow
one
of
these
pa-
tients
through
from
the
time
of
admission
to
the
end
of
convalescence,
to
see
at
what
time
the
principle
causing
the
appearance
of
the
clear
patches
first
appeared.
This
is
what
I
did
with
the
first
case
which
was
available.
The
first
day
I
isolated
from
the
bloody
stools
a
Shiga
dysentery
bacillus,
but
the
spreading
on
agar
of
a
broth
culture,
to
which
had
been
added
a
filtrate
from
the
faeces
of
the
same
sick
man,
gave
a
normal
growth.
The
same
experiment,
repeated
on
the
second
and
third
days,
was
equally
negative.
The
fourth
day,
as
on
the
preceding
days,
I
made
an
emulsion
with
a
few
drops
of
the
still
bloody
stools,
and
filtered
it
through
a
Chamberland
candle;
to
a
broth
culture
of
the
dysentery
bacil-
lus
isolated
the
first
day,
I
added
a
drop
of
the
filtrate;
then
I
spread
a
drop
of
this
mixture
on
agar.
I
placed
the
tube
of
broth
culture
and
the
agar
plate
in
an
incubator
at
37°.
It
was
the
end
of
the
afternoon,
in
what
was
then
the
mortu-
ary,
where
I
had
my
laboratory.
The
next
morning,
on
opening
the
incubator,
I
experienced
one
of
those
rare
moments
of
in-
tense
emotion
which
reward
the
research
worker
for
all
his pains:
at
the
first
glance
I
saw
that
the
broth
culture,
which
the
night
before
had
been
very
turbid,
was
perfectly
clear:
all