U
diversity
been
directed
in
cellulolytic ingested
symbiotic organisms Utilization
cellulose,
considerable
wood for
shipworms. comprehensively ruminant
ingesting
common.
Introduction
characterization
Characteristics
for
media a
analysis probes
eukaryotes,
dissolved
oligonucleotide
diagnostic
they
characterize
but
the
dense
Abstract
Prepared
Carolina,
Christian
variety
the
observed only
wood
many
serves
occur.
Many
exploitation
The
used,
at
The
concentrations
of
revealed
by
one Terenid
showed mammals
bacterial a
the
of
are
of
microorganisms
Among
and
oxygen
on linear
other for
shipworms
Associated utilization
stage
E.
occurrence Colwnbia,
media
These digestive
aspects
as although
cellulose from
of
organism A bereft
in
other
which Characterization
Microbial
Schlekar,
nitrogen-fixing
the
diagnostic
of
the
other
is
members probes,
bacterial
bivalves,
studied array
anaerobic consisted
a
this which
consortia.
the
molluscs
and necessary
under
of
shipworm’
consortia
bacterial
or
strains,
of
of
they
wood
organs, adaptation their of
termites
most
demands the
strain within
was
the pH.
residing SC
of
of
Department
and
wood Diversity,
are four
both
feed.
D-glucopyranose
forms
symbiotic
se
of bacterial
stage
or
in
with occurrence
supported of which
intensely-studied
conditions
as Microscopic
exhibit species
marine
are microelectrode
enzymes,
of
before
the The
shipwomis, the
a
e.g.,
strains
microscopic s
aerobic
bacteria
habitat
and
as
modified
cellulolytic A chief,
a in
was
similar that
flavobacteria
and
Gland
suite
a
hypothesis
cellulolytic was
the
the
the
their
nutritional
Summer community
radically
identification
within
wood-ingesting relationships
dwell used
of
were
specisilizaflon.
by
Gland
or
and
appendix,
however, was
of enriched within
from and
of Shipworm,
Environmental to
respective of perhaps
Waterbury gill
analysis
cellulolytic
utilize
to
anaerobic those
isolated. DeShayes
analysis, within
must
restricted
the
bacteria.
units
the
Lyrodzspedicellatus.
analysis
guide
of
1995 structure
that modified symbioses
all
nitrogen-fixing
source
group
shipworm,
DeShayes.
remain
under
of
it
rely a
organs
and
Microbial
linked
only,
bacteria
revealed between
of the
microbial
was
cellulolytic
the Desulfovibrio
Tentative
in
Wood terenid
media.
Well-characterized the
in for
on
to
enzymes.
beyond microscopic
is
shipworm.
called
enrichment
sulfate
situ morphological
not unclear
Health
source the
the
performed Lyroduspedicellatus
the
tested. by other
are the
aid
and
accepted bacteria
a gill
gill
For
is
(3(1-4)
hybridization
(1983) cellulolytic
those
bivalves, determination
symbiotic
Many
diversity
the
the
bacteria composed
in reducing
strains to
identification
microbial
of
Sciences,
despite
structure.
structure.
example,
Typically,
Enrichments
Community
the
Gland
delineate
well-characterized
nutrition.
This
between
spp.
examination
and
not This
colonies isolation
glycosidic
and
cellulolysis
until
is
commonly
has
features
of
the
without isolation
study
community of
conditions.
Additional
possible.
a
activity
bacterium
examples
largely
partners.
University prokaryotes
recently eukaryotic bacteria
Microelectrode
with
Oligonucleotide
DeShayes.
been
attention
regions
eukaryotic of
cellulose-
formed
was
of
was
proceeded
internal
bonds. that
identified; reveals a
of of
of
stage.
possible
known
initiated
have
that
include
Less in
occurs wood
allow to
of
on
host which
and
digest
South
The
all
a
on
as
is to
U
procedure.
storage supematant
tissue ethanol:fornialin placed
and
III) The
was
exogenous
microscope
II)
tissues
study. were MA.
shipworms laboratory
I) Materials
shipworm. isolated
using variety
Identification
detect Bacteroides whole
pedicellatus. groups
Oligonucleotide
systems
gain
bacterium,
methanogenic
of
cellulose
microbes,
organism.
the
presence
used
the
grinder
immersed
Cultured
into
insight
buffer95%
Organs DIC
in
appendix,
intact As
Microscopy
other animal
Select
Shipworms
Shipworms
presence
of
known bacteria
to
I
known
situ
individual
propose needed,
The
microorganisms
discarded,
diagnostic of breakdown had microscopy.
determine slides.
i.e., and
a
and Little
of symbionts.
group, Dr. into
organs
known
and true
In homogenates
animals
storage
Hybridization
in
methanogens bored
to
(v:v)
bacteria,
whether
respectively)
homogenized
to will
John probes of
Methods situ
a the ethanol,
be characterization
to is cellulolytic Sterile shipworms
harbor circulating
secondary
sulfate
sterile
known
fixative.
participatory
combine
from result
and nature
tests,
or hybridization
and
the
products
Waterbury
buffer
were
thought
will
The
Both or
would
presence
the
techniques
formed
and
bacteria,
freshly
in
e.g., Eppendorf
in
reducers,
not
of
about
and
maintained
pedicellatus)
be
outcome
pellet
was
consisted
this
broader
were molecular
Contents
maintained aerobic
consortia seawater
were
metabolism,
completely.
the
a that
determination support
selected
to specific
secondary tested
and calcareous at
sacrificed
in the
microbial
if
and
resuspended harbor will
namely
independently
are
carefully
Woods
isolated
were
cellulolysis.
members
phylogenetic
in understanding
of
centrifuge
and
diversity
of
generated
of
table
the
be
visually
is in
to
all
techniques
molecular
media.
at
were 40
employed
each microorganisms present
anaerobic
pieces
identify
employed hypothesis the
other e.g.,
Tubes
group
Hole -20°C
animals
community
cultures
tubes.
fIE2O°C)
mM
extracted
of
of
Gland
tube obtained
in
of
the
by
Non-specific
tubes
cellulolytic
procedures.
of Oceanographic
the
in
Tris Enrichments
in of
excised
were
until
microorganisms 1.5 specific
techniques
identity
F420 with
Wood
unidentified presence
the
were
of
enrichments to
were shipwonns.
bacteria Spirrilum
on
will
of
ml containing
that,
at
prevent at
the from
primary
the
spun
homogenates
within
DeShayes
from
appropriate
pH
autoflourescence.
all be
of separately
excised
from
pieces
microbial
phylogenetic
(i.e.,
or
initiation
instead
a
evaluated times
activity,
wood 7.5
for
is of
physiological Phase-contrast
media
a
50:50
and
the for
the
group
present
three
culture
cellulolysis.
sulfate
the
wood,
and
5
Institute,
will
containing
1
and
during
characterization miii
members introduction
in
digestive
of
and
ml
within
Gland
transferred
(v:v) enrichment
will
community
of
0.2%
presence
shipworms order
be will
a of
by squashed
90:10
at maintained
reducing
into the
that
the
single and
the performed.
also
the
the
3
solution
be
Woods
various
of
Nonidet
caecum.
hybridization
to
K,
metabolize
which
activity
two
of
system
physiological
shipworms Documentation
course outcome
made
microscopy
Deshayes
be
keep cellulolytic
of the
the
of
to
on
methods organ
used within
or
and
flagella
Hole, sterile the
of
organs.
in
of
on
the P40.
of
of
of
the
to
of
L.
these
the
the
the
to a
0
(Table
V)
examined.
Revsbech
through
were placed
pH
IV)
Bacteria
Low High
Delta
Arehaea
SRB
Flavobacteria
Gamma
Primer Probe
endosyinbiotic
Table
variety
according
made
G
G
2).
subdiv.
A
on
Isolations/Enrichments
Microelectrodes
Microelectrodes
1.
the
+
A
+
of
The number
subdiv.
(1989). an
C
Oligonulceotide
C
on
tissue
diagnostic
agar
hybridization
to
several
bacteria
Revsbech
of
plate
at
Dissolved
media
0.5
organs
oligonucleotide
below
mm
were
within
were
and
probes
procedure
increments.
by
oxygen
used
the
Jørgensen
the mechanically
used
microelectrode
used
to
shipworm
3
and
to
measure
followed
probes
in
isolate
I
pH
(1986).
used
in
situ
depth
lowering
(Table
Lyrodus
in
symbiotic distinguish
same distinguish
id.
id.
microelectrodes Bacreroides ii
the id.
ii
Purpose/Target (-i-)
vivo hybridization apparatus.
of
of A
of
of of
proffles methods
Control,
as freshly
methanogenic
1).
sulfate
sulfate
additional cellulolytic
dissolved
pedicellatus.
the
above
bacteria between
respective between
were of Cross-sectional
reducers universal
reducers
extracted
procedure
Nierzwicki-Bauer
oxygen
modified
cellulolytic
plotted
N 2 -fixing
from
archaea
bacteria
low
microelectrode
probe
shipworm
L.
concentration
and
for to
pedicellatus according
bacteria
and
identify bacteria,
each
measurements
high
bacteria
was
organ using
G
to
+
e.g.,
and
C a
-
preparation.
in
cultures,
probes
for
in individual
The
(Figure organisms relative
Microscopy
Results
physiological
tests
were
•
final
additional
homogenate.
before
anaerobic
bacteria.
at
-50
sterile from
Two
NB
SRB TYG
MCIY
the
pedicellatu.
Sea Media
Table
situ
approximately
75°C
gill
ul)
appendix were
concentration
examined
Water
to
re-applied,
tubes
Positive
The
Hybridization
Initial
removal
seawater
1)
Inoculates
abundance structure
with
homogenate 2.
three
for
eukaryotic
and
For
performed
were
carbon
hood
Media
slide
Resolution
and
Complete
10
inconsistent
microscopic
One activity intact
a inoculation
slide
controls
mm microscopically
observed
variety
placed
from
incubated
and
and
harbored
and five
used
source
received
were
of
of
prior
preparation,
shipworms drawn
was cilliates
thoroughly on
the
microorganisms the
of
hours
10mM
into
of to
of
were
checked pure
the liquid
agar
agar
agar
agar Matrix
incubation to
hood.
results. and
spread
in
isolate
fluorescence
of bacteria
examination
at an
0.3
into
an
application
bacteria.
squashes after
37°C
liquid
conducted
but
cultures
abundance for
sewed
ethanol-flame-sterilized
ml
were
and Three an
and
(agar/liquid)
across
homogenized. not for
endosytnbiotic
both
the
An
Na-lactate,
was
(Figure
NICO 2
SRB
re-initiated re-streaked
eukaryotic growth
by
as
initial
used
abundance
prepared
to
carbon
among
sealed
accidentally
varied
of to
a the
bacterial
on
determine
of media,
control.
NB
squashed
for
2).
cells
agar probe
sparged
microorganisms,
daily.
sources.
SRB
various each
agar
among and
F420
from
cellular
Bacteroides
at sulfate
m
spore
in
surface. bacteria
from spirochaetes/Spirrllu bioluminescent
shipworms For
aerobic Target bacteria
of cells.
application.
37°C
one order
plates
the As serum
media
exposed
wood
organs
5
agar fluorescence
the
the
ml several
organs
formers colonies
received phylogenetic The
tissue reducers
bacteria
material
in
heterotroph/
appendix
to
Shipworm
within
syringe. probes
plates,
for
particulate
bottles
type.
the
revealed
obtain
final
were
to
group
including
isolation
grinder
putatively of darL
This
light
appeared
0.3
L.
the Animals
occuned serum
applied sufficient
received
homogenized
was pure
or The
pedicellatus.
ml
shipworm
homogenate
slide at
differences
identity material
that the
of
room
exhibited Na-acetate,
ciliated
cultures. syringe
bottle
anaerobic
aerobic/anaerobic
anaerobic
aerobic/anaerobic
identified
Aerobic/anaerobic
aerobic
to
mid-gut
endospore-forming
on
were
was
contained
in
1
volume
shipworm
ml
the
temperature
plates,
and
received
washed,
was
Lyrodus
extracted
eukaryotes
was
of
in
in
gill
by
No Diagnostic
was
region.
pure
shipworm an
the
present
for
(e.g.,
they
sealed
several
slide
1-3
heated a
the
no
ml 25
0
vibrios,
both
the
plate dipolar,
container,
showed
opening.
streaking
with
amorphous, rods,
Component
Several
from
become
most
Enrichmencc
from
agar
anaerobic hour.
Respiration
Micro-Electrode
flourescence
Designation:
Gland Appendix
amp1e prepared
Table
effectively
fluorescent
flavobacteria
middle
were
the
whereas
samples
control
bottles.
higher
exhibited
below
and
round,
part,
appendix
7.6
Turbidity
Every
A
small
treated
no
Aerobic 3.
bulbous
highly
A apparent
was panel)
Anaerobic non-motile
to strong,
and within
from
conditions heartbeat
growth. universal Results
(able
growth
agar
at
organism loose cells
(-)
8.0,
The
raised, continued
captured
cocci,
cells
attempted,
organ
025
anaerobic
a
=
with
pigmented
probe
sample.
the and enrichments
broad
terminations,
no content
and
increased
lactate
putrescence-like
exhibited
four
until
Analysis
colonies 3).
second
were
occurred
of
flourescence.
Microscopic
and
gill
Archae
and
that
Primer
was flavobacteria
enrichments
did
curved
in
immediately
showed For
on
once
and
array
two
throughout
(-)
(—)
structure
situ
bottle embedded
irregularly
Clearly
glossy
conditions
yielded integration
not
was
but
observed
film.
example,
colonies
seven
in
that
weeks
a
the
C
291
within
of rods.
differ the
hybridization
range
the
on probed,
the
exhibited
vibrios,
(45°C)
probe
microbial
No
were
+
colonies
visible
catalyst
(Figure SWC
better
examination
days, SRB
=
and
on
most
after
odor
(Figure
Aerobic
in
detectable
below the
significantly
within
fluorescence
of
two were
shaned
were
universal
TYG
initially
the L0G+ë
appendix had
i.e.,
shapes,
Na-lactate
fluoresced
plates
analysis,
respectively.
separation
and
intense
the
emanated
fluorescent
days.
more 3,
to
was present, (-)
(—)
organism
not
forms,
the
passed
eukaryotic
gill
3,
plates
initial
with
rods.
middle
long flattened,
enrichments
flourescence
bottom
achieved. yielded
not difficult tissue
Primer of
abundance including
structure,
However,
and
from
which
oligonucleotide
from
was
constituents
narrow including
replaced exhibited
and
inoculation.
SevtrI when
with
from
of
completely
row,
used
widespread cells
surface.
colonies. row,
A
A
the observed
a
Arch
Na-acetate
structures,
amorphous,
the
to
colors
proceeded
variety
at
the variety
applied
(37°C)
Many
left
for
short
rods.
were on
separate. appendix,
shipworm
0.5 some
and
(—)
exterior
fa-rm!
right
+
upon
cocci,
anaerobic
rich,
MCTY
micro-electrode
panel),
second
Oxygen
from
ranging
diversity;
through
motile visible
of
of
Colony
in
did
of
colony
panel)
fluorescence.
to
resealing
rf
probes
amorphous
although
serum
the colony
non-motile the
for
microbes
environment.
the
spreading
Preparation
the
integration
not
lw’+r
Lyrodus
brood
agar
13
(-)
(—)
re-streaked
rods,
appendix
was in
approximately
Gas
from
(Figure gland
probes.
the
initial
morphotypes
fluoresce
forms
bottles
cell
applied
gland
plates forms.
the detected
pouch,
this
tissue.
Pak
long
(—)
÷
p
yellow
were
types
pedicellatu. sample
growth.
analysis.
colonies.
enrichment
++ rods
did
Gas
Many 3,
samples was
The
upon
relative
of
yielded samples.
to
non-motile
=
middle
For
plates
at
present
not
(-)
(-)
exhibited
pH
plates
included
detectable
Pak
with samples
not
in
to
all,
but
ranged
one
the
ranged
Re-
the
red.
to
that
not
(-)
(—) row,
in
Flay
++ (—)
fl
0
polar
dilutions
shaped. Logistical
rod-shaped,
from
catalase
(Table the refractile pasteurization but
5). These
bulbous
amorphous morphotypes,
plates, were morphologically
three Isolations
Table
from
with
remained bottles
Yellow
the
Long-Curved
Designation
SRB nature
not
Pale Growth,
MCIY flagellum
acetate
not
pure cells
6). Isolated both
One the
The
i.e., on
Pure
4.
Microscopic
positive
Isolated Vibrio
White
ends, (i.e.,
produced areas achieved
Cluster restraints
of
cellulose.
acetate
Characteristics
cultures,
motile, were active
pure the
pale carbon
(Table
agar the
albeit cultures
emichment,
iO
or
in one
was
at
colonies
isolated
refractile
colonies
white
(Table
culture
exhibit immotile,
the
various
plate distinct
under
to enrichment
yellow,
4).
despite
minor, 3.8 sources.
prevented
and
observed
two
MCTY
yellow-pigmented
examination iO SRB
They SWC
SWC grown colonies
Microscopic and
inoculations
scored
5),
colonies
were
that anaerobic
fluorescence.
on
original
points
colonies
from
as
Media
areas.
occurred
of
shake
the
glossy,
and
could 1.4
gram
agar
agar
SWC
agar was
on
Agar
formed
colonies
under
present
any
because
consistent
positive
mM
the
did
were
SWC
along
plate
plate a
tube
plate
always These negative,
not
concentration). agar
showed further
deeps
on
eukaryotic
first
conditions and
not
DIC
(Table
sulfide,
on
examination
amorphous,
composed grow
within
SWC
agar
isolated
the
of
plates
circular grow
cellobiose,
re-streak
for
I
motile colonies
microscopy.
were consisted
occurrence
its
characterization.
Aerobic/Anaerob
length the
plates
4). oxidase
the
anaerobically,
were
respectively,
slow
four
on
and
for
flagellate
cells
prepared Anaerobic
rods oxidase These from
and
Aerobic
Aerobic
Aerobic
cellulose
of
of flattened
revealed (Fig. repeatedly (Table
exhibited at
days
one development.
The
long carboxy-methyl
of
ic
negative, to
initial
were least
the
the
of
gram
a
be
on
7).
in
colonies
and
mixed isolated
other
dark from
(Fig.
shipworm,
5,
rapidly-motile
thirteen
MCTY
gram
all
indicating
or
enrichments nor
curved DIC white
Figure
negative
two catalase
re-streaked,
cellulose SRB
and
cells 4).
the
pale
did
culture.
negative,
microscopy
colonies
different
were
colonies
days. lactate
agar catalase Interestingly,
bright
Lens-shaped,
Na-lactate rods, they
that 5
white,
Flat,
Lyrodus
cellulose,
Colony
Small,
upper tests
that
cells
brown
breakdown-products
Flat, plates.
spreading,
exhibited
on
form
The
sometimes
vibrios.
diffuse,
yellow
enrichment,
on sulfate
but oxidase
colony
flattened,
were
positive
was
pale
agar
(Table
left,;
glossy,
amorphous,
did re-streaked
lactate
pedicellatus.
pure
and agar
spores
Description
Three
and
obtained
white
plates
irregularly
the
pigmentation
not
Figure
was
light
non
spreading,
A
lens- 5).
and
white
deep cultures
xylase,
(Table
flagellate
with
and single
raised, and
reveal
upon
reduced
but
yielded
brown
6).
acetate not
C)
CD
other
spp.
shipworm. homogenization,
because
environment marine
example,
Caution report
lactate putatively
that
enrichment.
these,
share
any
activity, grown
isolated
either
moved
bacteria
isolated
archaebacteria,
was
anaerobic were microorganisms
Discussion
ND
SP 1 :
SRB
Long-Curved
Pale
Yellow pedicellatus.
Designation
Table5.
is
formed
further
participants
obtained
single
white
the
Vibrio
of
putative isolated
surviving
as
and
organisms
on
The
via
the
Waterbury
must
Enrichment
Cluster
in
from
an from
16
it
a
characteristics
MCrY
identified
polar
absence conditions
spiraled
the
is
This
a
Cell
organisms
conclusions
anaerobic,
sulfate-reducing flattened,
s
within
The
single
difficult
rRNA
be
from
aerobic
the
present in
identification
flagellum
characteristics
members
at
still
it
exercised
rods
Vibrio
Long
pure
of
transparent featuring
Long
with
Curved
identity
Cell
within
(Harris
Gland
agar
is
he
er
in
or
polar
cellulolytic
its
an
in
based
techniques
reasonable
does
bulbous
al.
pale-white
irregularly-shaped
expense using
to
acetate
rods,
long
culture,
agar
were
study
sulfate-reducing
burrow, situ
anaerobic
plates.
Description
rods,
about
of
as
distinguish the
of
(1983)
non-refractile
flagellum,
of
1993).
of
not
when
determined
on
elements
always
the
hybridization)
plates.
straight
DeShayes
thoroughly
several
shipworm
the
that
bacteria
is
sometimes
the
end
(Waddle
its
clarify
its
enrichments
cellulolyitc
Further
of
possible
from
shows
symbiotic
curved
isolated and
to
3
matches
considering
vibrio
colonies
identity.
sulfate
Given
by-products
subclass,
In
accept
rod
types
between
and in
pure
by
its
appears
situ
of
anaerobic
situ
characterization Lyodrus
and
(Waterbury
rod
DIC
rinsed bacterium
shape,
only
that
status,
exhibited and reducing
L.
Motility
of
these hybridization
that colonies
remains
of
systems
rod
None (+):
hybridization
Bak
yielded
pedicellarus.
that
microscopic
and
enrichment
characterized
the this
for
resident
(+)
the
to
(-)
(-)
Desulfovibrio
of
identified
with
SPt
single,
characteristics
i.e.,
pedicellatus.
formed
1992).
be
woodworm
flavobacteria.
the
conditions
of
isolate
role
celluloysis.
culture.
isolated
morphological
unclear.
may
isolated
Desulfovibrio
a
et
sterile
the
transient
vibrio
variety
of
and polar
al.
analysis
Gram
other
To
(i.e.,
tentatively
support
yellow
would
media.
Stain
SRBs
by
demonstrated
1983).
This
transient
seawater
a
from
Three
isolated
my
(-) from
(-)
(-)
Waterbury
flagellum,
within
cellulolytic,
of
creates demonstration
spp.
bacteria
vs.
Although
is
within knowledge,
bacterium
be
colonies
organisms
the
At
any
the
resident.
the
The
other
did
pleomorphy,
spp.
necessary
indicated
microorganisms
role all least
from
prior
irregularly-shaped
an
terenid
Test shipworm
Ozidase
originate
only
isolated
the
organ
and
This pure
et
a
aerobic
that
only
and
one
nitrogen
the broad
(-)
to
(-)
al.
was
shipworm.
under
bacterium
Identification its
this
of
bivalve.
cultures
bacteria
Desulfovibrio
the
before
SRB
the
pure
(1983).
systems,
four
presence
in
gram
cellulolytic
within
Lyrodus
appearing
diversity
is
long presence
aerobic
this
fixing
the
culture
bacteria
Test
Catalase
making
(-),
within
were
was
study
rod
Of
first
For
the
ND
and
(+)
(+)
(-I-)
rod
in
of
and
of of
0
Waterbury,
WiddleandBak
Revsbech,
Revsbech,
Harris,
References
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culture,
were
conditions
hindgut
of
outside
observed
documentation
observed
Teredinidae).
bacterium
293
ecology.
use
invertebrates: J.M.
To
In
including
the
(Joy
and
within
N.P., N.P.
addition
of exhibits
J
summarize,
-
scope
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indeed
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Sabi,
oxygen
within
and
C.B.
1989.
:
1992.
cultured the
of
what
of
Plenum,
personal
to
external
The
much
Marshall,
B.B.
shipworm.
this
such
Science
A
Calloway,
microsensors.
the
the
Diffusion
a
appears
synthesis.
presence,
number
study,
variety
from
Jørgensen.
shipworm,
of
symbiotic
NY.
morphological
communication).
the
221:
K.C.
the
their
to
Although
microbiota
and
of
characteristics
of
9
be
nature,
Microbial
Gland
1401-1403.
bacteria
microorganisms,
(Ed.),
organisms.
presence
R.D.
L.
J.
Desulfovibrio
1986.
pedicellarus.
Microbiolo.
of
and
Turner.
“Advances
characterization
resemblance
of
DeShayes
described,
Microelectrodes:
Ecol.
is
role
L.
of
nonetheless
One
pedicellatus,
microbial
of
1983.
spp.
25:
Meth.
ciliate
both
gut
Four
in
in
several
195
to
Microbial
The
shipworms
microflora
A
prokaryotic
a
9:
of
bacteria that
-
protist
cellulolyitc notable
communities
identity
231.
eukaryotic
111-122.
remain
eukaryotic
Their
survived
occurring
Ecology”,
were
because
in
use
(Bivalvia:
of
unclear.
and
aquatic
the
nitrogen-fixing
organisms
in
brought
under
organisms
determined
eukaryotic,
microbial
remaining
in
of
Vol.
the
anaerobic the
into
termite
9.
absence
is
were
p.
pure by
fl
Lyrodus
pedicellarus.
Figure
as
single
Figure
features
pedicellatus.
non-refractile
Figure
shipworm
Figure
bulbous
White
long
panel
plates.
Figure
NOTE:
homogenate
Figure
bottle
middle
show
show homogenated
Figure
freshly
Figure
the
Figure
sacraficed
structure
Figure
a
carbon
gill
rods
polar
shows
extraneous
flourescent
8.
7b. cells. after
7a.
6.
5.
4.
Original
common
pedicellarus..
3.
panel),
2.
ends,
sacraficed
1.
structure
FIGURE
TOP:
designated
DIC
Lyrodus
Phase
Figure
Legends
of
Phase Phase
Phase
DIC
source.
flagellum
Phase
thirteen
specimen,
curved
of
Isolate
the
Isolate
features
i.e.,
samples
micrograph
Phase
and
the
micrograph
innoculate
contrast
contrast shipworm.
contrast
to
contrast
6.
material
of contrast
cells:
8
middle
pedicellatus.
specimen,
BOETOM:
shipworm
all
the
days.
IS
rods
is
the
Phase
is
“Pale
extending
common
contrast
and
Culture
designated
cells.
designated
OUT
flavobacterium of
Archae
shipworm,
micrograph
designated
micrograph
micrographs
micmgraphs
shipworm
SRB
micrographs
of
that
squashed
White”.
contrast
of
was
Lyrodus
of
OF
upper
a Lyrodus and
micrograph
was
non-specifically
a
DIC
to
pure
from serum
291
homogenized
pure
ORDER
Isolate
all
a
“Pale
squashed
“Yellow
isolated
LyroduspedicellatzLs.
right
micrograph
Note
micrograph
probe
on
(Lyrodus pedicellarus.
culture
as
cells.
each
of
of
culture
bottle
of
pedicellatus.
of
probe
of
“Yellow
a
White”.
bacteria
microbiota
is
bacteria
panel.
microscope
the
oligonucleotide
cell
eukaryotic
designated
of
(middle
Cluster”.
under
on
isolated
was
presence
isolated
a
(bottom
pedicellatus)
shipworm
pure
a
bound
of
of
Cluster”,
from
Note
microscope
innocluated
and
sulfate
a
cells
Sample
row,
present
culture
pure
from
Note
organisms
ekaryotic
slide.
from
row,
“Pale
isolated
the
the
of
in
left
(Lyroduspedicellatus).
reducing
non-refractile
culture
probe
presence
probe.
and
the
Sample
SRB-lactate
the
right
was
the
in
White”.
gill
isolated
panel),
slide.
with
acetate
shipworm
presence lower
colonies
shipworm
organisms
obtained
structure.
flourescence
panel).
observed
isolated
anaerobically-prepared
was
conditions
of
from
right
Note
SRB
probe
non-refractile
obtained
isolation.
features
on
of
The
Lyrodus
from
from
Lyrodus
the
within
panel
Three observed
the
cells
SWC
enrichment
(middle
other
applied
shipworm
using
presence
a
the
exhibiting
of
from
freshly
shows
agar
panels
the
Upper
Note
e
Pale
panels
lactate
within
row,
gill
to
a
the
of left /
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