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Bone-Boring Worms: Characterizing the Morphology, Rate, and Method of

Bioerosion by Osedax mucofloris (Annelida, Siboglinidae)

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Reference: Biol. Bull. 221: 307–316. (December 2011)
© 2011 Marine Biological Laboratory
Bone-Boring Worms: Characterizing the Morphology,
Rate, and Method of Bioerosion by Osedax mucofloris
(Annelida, Siboglinidae)
NICHOLAS D. HIGGS1,2,* ADRIAN G. GLOVER2 THOMAS G. DAHLGREN3,¶
AND CRISPIN T. S. LITTLE1
1
School of Earth & Environment, University of Leeds, Leeds, LS2 9JT, UK; 2Zoology Department,
Natural History Museum, Cromwell Road, London, SW7 5BD, UK; 3Department of Zoology, Göteborg
University, Box 463, 405 30 Göteborg, Sweden
Abstract. Osedax worms possess unique “root” tissues
that they use to bore into bones on the seafloor, but details
of the boring pattern and processes are poorly understood.
Here we use X-ray micro-computed tomography to inves-
tigate the borings of Osedax mucofloris in bones of the
minke whale (Balaenoptera acutorostrata), quantitatively
detailing their morphological characteristics for the first
time. Comparative thin-sections of the borings reveal how
the bone is eroded at the sub-millimeter level. On the basis
of these results we hypothesize a model of boring that is
dependent on the density and microstructure of the bone.
We also present evidence of acidic mucopolysaccharides in
the mucus of the root tissue, and hypothesize that this plays
an important role in the boring mechanism. We discuss the
utility of these new data in evaluating Osedax trace fossils
and their relevance for O. mucofloris ecology. Measured
rates of bone erosion (6% per year) and evidence of en-
hanced sulfide release from the borings indicate that Osedax
worms are important habitat modifiers in whale-fall com-
munities.
Introduction
The arrival of a whale carcass on the seafloor represents
a large energy input to the local benthos and can support
dense communities long after the soft tissues have been
*To whom correspondence should be addressed. E-mail: n.higgs@
nhm.ac.uk
¶
Current address: Uni Environment, Postboks 7810, N-5020 Bergen,
Norway.
Abbreviations: AMPs, acidic mucopolysaccharides; CT, X-ray micro-
computed tomography.
307
consumed (Smith, 2006). The whale skeletons that remain
host some of the most species-rich hard-substrate commu-
nities in the deep sea, reflecting the high trophic diversity
found among their fauna (Baco and Smith, 2003). One of
the more unusual adaptations found at these “whale-fall”
communities is in polychaete worms of the genus Osedax
(Annelida, Siboglinidae), which use root tissues containing
bacterial symbionts to derive nutrition through the excava-
tion of the bones (Rouse et al., 2004). Much of the work
carried out on these worms has focused on the nature of
their bacterial symbiosis and unusual reproductive traits
(e.g., Goffredi et al., 2007; Rouse et al., 2008; Verna et al.,
2010), but little attention has been paid to how these worms
interact with their substrate. In particular, there is a paucity
of information on the morphology of their borings, how they
bore into the bone, and how this affects the longevity of
whale bones as substrates.
Osedax species are the only members of the Siboglinidae
family that bore into hard substrates, but this habit is wide-
spread and ancient among the polychaetes (Taylor and
Wilson, 2003). Several families, such as the Spionidae and
the Cirratulidae, contain well-studied boring species, and at
least seven other families include species that also bore into
hard substrates such as calcareous rock and shells (Warme,
1975; Hutchings, 2008). In doing so they create a stable
domicile and center for foraging. Polychaete borings have
existed in hard substrates for over 350 million years (Cam-
eron, 1969), suggesting that this lifestyle was an early
adaptation that has evolved several times among the group.
There has been some debate as to whether polychaete
boring is carried out by chemical means (Haigler, 1969) or
mechanically using modified chaetae (Hannerz, 1956). Af-
308 N. D. HIGGS ET AL.

Figure 1. Location of the sample site with the 100-m isobath highlighted in dark grey. Redrawn after
Dahlgren et al. (2006) with permission from CBM—Cahier de Biologie Marine/Station Biologique de Roscoff,
and Palm et al. (2004) with permission from Elsevier.

ter reviewing the literature and conducting experiments,
Van der Pers (1978) established that a combination of both
mechanisms is important for boring in polychaetes. Despite
this scrutiny, the identity of the chemical agent responsible
for boring remains unknown (Hutchings, 2008), although
Dorsett (1961) suggested that the mucous tubes of some
polychaetes might contain acidic mucopolysaccharides
(AMPs) that aid in the process.
The boring lifestyle affords moderate protection for
Osedax mucofloris Glover et al., 2005, which is capable of
retracting into the bone when threatened (Glover et al.,
2005). It also provides a means of obtaining nutrition:
Osedax root tissues produce proteolytic enzymes that can
break down the protein matrix of the bone (Goffredi et al.,
2007). While whale carcasses are probably the preferred
habitat for Osedax (Glover et al., 2008), the ability of the
worms to live on ungulate bones suggests that they may not
be restricted to cetacean remains (Jones et al., 2008; Vri-
jenhoek et al., 2008).
The evolutionary history of the species-rich Osedax ge-
nus remains ambiguous. A recent molecular analysis by
Vrijenhoek et al. (2009) resulted in two hypothetical sce-
narios in which Osedax split from their siboglinid relatives
either 45 MYA (concurrent with the evolution of large
cetaceans) or 135 MYA when large marine reptile carcasses
could have provided suitable habitat. The boring traces that
Osedax produce in whale (and other mammal) bones pro-
vide a way to test these hypotheses in the fossil record. So
far two studies have claimed to find such structures in whale
bones: one of a bone from from the early Oligocene (⬃30
MYA; Kiel et al., 2010) and one from the Lower Pliocene
(5.3–3.6 MYA; Muñiz et al., 2010). However, information
about the morphology of living species that is needed to
evaluate these claims is lacking.
Gathering accurate details of Osedax boring is difficult
because it is usually necessary to destroy the bone to ana-
lyze the root tissues. Higgs et al. (2010) showed that X-ray
micro-computed tomography (CT) is a useful tool for in-
vestigating the boring behavior of Osedax mucofloris, but
failed to detect any individual borings, only a large cavity of
merged borings. Using CT in concert with direct observa-
tion, we present the first detailed morphological analysis of
 OSEDAX BORINGS IN WHALE BONES
Table 1
Details of the bone samples used for this study, which were colonized by
Osedax mucofloris
Max. Osedax Borehole
Exposure density diameter Max. boring
Bone (months) (cm⫺2) (mm) depth (mm)
Ulna 8 2.40 0.28⫺0.74
Radius
upper surface 12 1.96 0.41⫺0.55
lower surface 12 2.45 0.38⫺0.54
Vertebral process
1 10 1.29 0.43⫺0.45
2 55 — 0.34⫺0.58
borings (with quantitative data) for any modern Osedax
species and provide new insights into the method of boring
and the factors that influence it. These data also yield useful
information about the growth, nutrition, and ecology of this
enigmatic group of animals.
Materials and Methods
Study site and sample collection
All samples were collected from the Kosterfjord in Sweden,
a 250-m-deep, 48-km-long trench bordered by the Swedish
mainland to the east and the Koster Islands to the west (Fig. 1).
At the study site in a water depth of 125 m, bottom water
temperatures vary between 4.8 and 7.5 °C, with salinities of
34.3–34.7 PSU, and oxygen levels of 4.7– 6.3 ml l–1 (Dahlgren
et al., 2006). Sediment traps deployed at 100 –130 m in the
Kosterfjord measured a settling rate of 3.14 g m–2 d–1 for
particulate matter (Palm et al., 2004).
In May 2008 we deployed, using a remotely operated
vehicle (ROV), a package of dissected minke whale
(Balaenoptera acutorostrata Lacépède, 1804) bones, in-
cluding an ulna and a radius, near (⬍10 m) to a whale
skeleton at 125 m as a part of an ongoing study into
whale-fall communities (Dahlgren et al., 2006). The ulna
from the bone package was collected with a ROV after 8
months, and the rest of the bone package was collected after
a year, providing two time-points to compare the rates of
bone destruction by Osedax mucofloris. The ulna and radius
were fixed in 10% seawater formalin and later stored in 70%
ethanol. The bones were cut into sections for analysis.
In addition, bones from the original skeleton (deployed in
2003) were utilized in this study (Table 1). In August 2004 a
vertebra was collected from this experimentally deployed
minke whale carcass (Dahlgren et al., 2006). A trapezoid
section of this process (VP1), which hosted Osedax speci-
mens, was sawn off and fixed in 10% seawater formalin.
Another vertebra was collected in May 2008 from the same
carcass, and a whole process (VP2) was sawn off and fixed
in 10% seawater formalin. These vertebral bones have a
309
different bone structure than the ulna and radius, so they
allow us to test for the effect of bone structure on O.
mucofloris boring morphology.
X-ray micro-computed tomography
Borings in the bones were imaged and analyzed using
X-ray micro-computed tomography (CT). Bone samples
2.63
with specimens of Osedax mucofloris in situ were scanned
4.32 using a Metris X-Tek HMX ST 225 CT system at the
4.78 Natural History Museum, London. Data volumes were con-
structed using CT PRO ver. 2.1 (Metris X-Tek, UK) and
1.65 analyzed using VG Studio Max 2.0 (Volume Graphics,
2.40
Heidelberg, Germany). These volumes are made up of vox-
els (the three-dimensional equivalent of image pixels) that
determine the resolution of the digital model. All bones
were scanned to provide a resolution of 65 ␮m or greater.
In the first stage of CT analysis the bone tissue was
separated from non-bone material (air, marrow, preservative
solution) on the basis of the density properties of each
material, a process known a thresholding (see Roche et al.,
2010, for more detail). Osedax borings were identified as
continuous cavities in modified bone tissue. Only cavities
that had a single hole leading to the surface were assumed
to be an individual Osedax boring. Direct measurements of
volume, surface area, maximum depth of penetration, and
width were made for individual borings (i.e., those not
touching another boring) using tools of the VG Studio Max
software (Fig. 2). The total volume of eroded bone was
measured using the SPIERS software package, ver. 2.09, by
comparing the total number of voxels that make up all
cavities and those that make up the bone tissue.
The relatively low density of O. mucofloris tissues in
comparison to bone inhibited their detection using CT.
Voids in the bone tissue were assumed to be the result of O.
mucofloris boring activities when coincidental with visually
identified areas occupied by the worm. To test this assump-
tion, a thin slice of Osedax-containing bone from the ulna
was post-fixed in osmium tetroxide and CT scanned. The
osmium tetroxide absorbed high-energy X-rays, allowing
visualization of the soft tissues to which it was attached, and
showed that the root tissues (imaged as a cloudy texture) did
fill the entire void in the bone (Fig. 3).
Analysis of the bone-tissue interface
An individual specimen of Osedax mucofloris was care-
fully dissected, intact, from a freshly recovered minke
whale vertebra that had been on the seafloor for 6.5 years. A
thick mucous layer that adhered to the bone, but remained
attached to the specimen when it was removed, heavily
covered the specimen. The O. mucofloris specimen was
stained with alcian blue at pH 2.5 for 30 min, then washed
for 15 min in distilled water and fixed in absolute ethanol to
test for the presence of AMPs (according to Curran, 1964).
310
Figure 2. CT reconstruction of Osedax mucofloris borings. (A) Trans-
verse section of the radius showing isolation of a boring (outlined in white)
and measurement of longest axes. (B) The reconstructed boring with a part
of the bone removed. (C) The isolated boring used for analysis.
To investigate the effect of this mucus on the bone, we
dissected a piece of bone from the root tissue of the spec-
imen and examined it under a Leica DM5000 high-power
light microscope to look for signs of erosion. We also cut a
3-cm ⫻ 2-cm ⫻ 1-cm section of bone out of the radius,
which contained O. mucofloris borings. The bone section
was oven-dried at 35 °C for 1 week and then embedded in
resin. This block was then ground down to a 100-␮m
petrographic thin section, allowing us to investigate how the
O. mucofloris roots had interacted with the bone tissue to
form the borings.
Results
Bone colonization
On collection, the ulna appeared structurally intact after 8
months on the seafloor, with the exception of small holes
where the Osedax mucofloris tubes penetrated the bone
surface (Higgs et al., 2010). The upper surface of the ulna
that projected above the seafloor was densely populated by
O. mucofloris (density reaching 2.40 cm–2), with greatest
concentration located on the mid-section of the bone surface
(Table 1). There were distinct patches of smaller specimens
located on the central surface of the bone and near the wrist
joint (Higgs et al., 2010).
The second bone to be collected from the experimental
N. D. HIGGS ET AL.
Figure 3. Test to visualize Osedax mucofloris root tissues in the
borings. (A) A slice of bone with an O. mucofloris boring (right). (B) The
same sample after staining with osmium tetroxide. O. mucofloris tissues
show up as a cloudy texture in the boring.
bone package was the radius, recovered in the spring. After
a full year of exposure on the seafloor, the radius was in a
similar condition to the ulna, with numerous individuals of
O. mucofloris again concentrated on the mid-section of the
upper surface of the bone (Fig. 4A, B). There were no living
specimens on the underside of this bone, but there were
Osedax borings (Fig. 4C, D), which were more numerous
and densely spaced than on the upper surface (Table 1),
indicating that the bone had probably rolled over and smoth-
ered the Osedax population at some point during the 12-
month period. Holes on the surface of the bone, where
Osedax tubes penetrated, were stained by a halo of black
precipitate (Fig. 4D) that sometimes appeared grey because
of bacteria growing on these areas (Fig. 4B). These halos
corresponded precisely with the extent of the root tissue
underneath the surface layer of bone (Fig. 4C, E).
The surface of the small trapezoid section of vertebral pro-
cess (VP1) was intact and solid, except for areas where two O.
mucofloris specimens had been dissected out during a previous
study. A thin (ⱕ0.3-mm) layer of bone covered the root tissues
of the specimens embedded in the bone. Eleven individuals of
O. mucofloris were found on the dorsal side and three on the
ventral side of this bone. This equates to densities of 1.29 cm–2
and 0.35 cm–2 for the dorsal and ventral sides, respectively.
The whole vertebral process (VP2) was heavily degraded and
only small, elevated patches of cortical bone remained. Three
 OSEDAX BORINGS IN WHALE BONES 311

Figure 4. Osedax mucofloris borings in the radius. (A) The whole radius (upper side), with dotted lines
indicating the section that was CT scanned. (B) Close-up of the section indicated in A. (C) CT scan showing the
lower surface. Part of the bone has been sliced away to reveal the outline of the borings. (D) Lower surface of
the radius showing bore-holes (some merged) surrounded by halos of black precipitate. (E) CT scan of the upper
surface, with some reconstructed borings visible in green, where part of the bone has been made transparent. (F)
Thin section of the bone showing Osedax borings and microborings (arrows) in proflie. Abbreviations: Bo, whale
bone; Ob, Osedax boring.
312 N. D. HIGGS ET AL.
Figure 5. Quantitative morphology of individual Osedax mucofloris
borings in the radius. (A) Changes of width (circles) and depth (squares) of
borings with volume. Regression lines for width (solid) and depth (dotted)
are defined by the equations y ⫽ 2.1001x0.3565 (R2 ⫽ 0.94) and y ⫽
0.7443x0.3244 (R2 ⫽ 0.81) respectively. (B) Changes in the surface area of
the borings with volume. Solid line shows regression line defined by the
equation y ⫽ 8.8148x0.7705 (R2 ⫽ 0.82).
individuals of O. mucofloris were observed living on this bone
when it was collected.
The morphology of Osedax mucofloris borings
X-ray micro-computed tomography (CT) scanning of the
ulna revealed wide, shallow, subsurface cavities created by
the merger of numerous borings, but failed to show any
individual borings (Higgs et al., 2010). This prevented an
accurate description of the Osedax mucofloris borings from
that bone. In contrast, the borings on the upper part of the
radius (Fig. 4A, B) were less densely spaced (1.96 cm–2)
than in the ulna (2.40 cm–2), and 26 individual borings could
be defined from the CT scanned section, although many
were merged. The O. mucofloris borings were approxi-
mately hemi-ellipsoidal in shape, flattened at the top where
the Osedax root was covered by a thin layer of bone, and
rounded toward a point of maximum depth inside the bone
(Fig. 4E). A few borings were more conical in profile,
though all were widest just under the surface and narrowed
with increasing depth into the bone. In total, the borings
accounted for about 6.0% ⫾ 0.8% of the volume of the
scanned section of bone.
Figure 6. Osedax mucofloris borings in vertebral process 1 (VP1). (A)
CT scan of bone sample with surface layer of bone made transparent to
reveal subsurface cavities. Dotted white line shows location of B. (B)
Cross-section through bone sample showing subsurface cavities; note thin
bone layer covering cavities.
The borings show a consistent hemi-ellipsoidal morphol-
ogy over a range of sizes (Figs. 4E, 5A). The high R2 values
of the curves in Figure 5A show how little deviation there is
from the model hemi-ellipsoid morphology in this bone.
The close affinity in the shape of curves in Figure 5A show
that vertical and horizontal borings increase proportionally,
although the width is always greater than the depth of
boring. The ratio of surface area to volume of the borings
also decreases according to an inverse power function (Fig.
5B) as the borings become larger, as would be expected for
a hemi-ellipsoid morphology.
Borings in the vertebral processes differed from those
seen in the limb bones. None of the borings penetrated
deeper than 1.65 mm in the trapezoid process piece of bone
(VP1), but instead spread laterally to form shallow subsur-
face cavities (Fig. 6), restricted to the densest part of the
bone. Similarly, several empty borings identified in VP2
were shallow and laterally expansive (Fig. 7), despite being
exposed to Osedax for much longer than VP1. Borings
corresponding to the locations of the three living specimens
on VP2 could not be detected in CT scans because their
roots were highly dendritic. This bone was exposed to
Osedax for 55 months, but the depth of penetration was
similar to that in the ulna, which was exposed for only 8
months (Table 1).
 OSEDAX BORINGS IN WHALE BONES
Figure 7. Osedax mucofloris borings in vertebral process 2 (VP2). (A)
CT scan of bone sample showing remaining cortical bone punctuated by
bore holes (numbered 1–7). Dotted white line shows location of B. (B)
Cross-section through bone sample showing borings with entrance holes
(numbered arrows).
Bone-tissue interface
Osedax mucofloris is named for the tube of mucus that
surrounds the trunk of the body (Fig. 8A). The outer mucus
that covered an extracted specimen also enveloped the root
tissues inside the bone. This mucus stained positively with
alcian blue, indicating the presence of acidic mucopolysac-
charides (AMPs) (Fig. 8C). Although root tissues and bone
were also directly exposed to the stain, both failed to bind
with it, showing specificity to the mucus (Fig. 8D). Detailed
investigation of the bone piece attached to the root revealed
extensive microborings in the shape of branching grooves
that were etched into the bone’s surface (Fig. 8E). These
microborings are smaller in scale (⬃10-␮m wide) than the
borings produced by an individual O. mucofloris. Microbor-
ings were also observed on the thin section through an O.
mucofloris boring (Fig. 4F); however, these were restricted
to the thin layer of bone covering the root tissue.
The thin section of O. mucofloris borings (Fig. 4F) bisected
a large boring longitudinally and another smaller one at an
oblique angle, exposing the hemi-ellipsoidal shape of the large
boring. Inside the borings the bone tissue has been mostly
eroded away, but they are not completely empty. Instead, thin
rinds of trabecular bone remain, outlining the structure of the
bone before it was eroded. These rinds are contiguous with the
uneroded trabeculae, and at the edge of the borings an “erosion
313
Figure 8. Osedax mucofloris. (A) In situ, emerging from a bone
surface with the trunk surrounded by the mucous tube (arrows). (B) Plan
view with bone surface removed to reveal elongated root tissue in a whale
vertebra. (C) Dissected specimen exposing the mucous lining (stained with
alcian blue) and attached piece of bone. Dotted white line shows boundary
between bone, root tissue, and the trunk of the specimen. (D) Micro-
borings on the surface of bone attached to root tissue. (E) Detail of the
exposed root tissues (arrow) from C which did not positively stain. Ab-
breviations: Bo, whale bone; O, ovisac; Rt, root tissue; Tr, trunk.
front” can be seen where the bone tissue has been eroded
within the trabecular struts of the bone.
Discussion
X-ray micro-computed tomography has recently been
applied to Osedax (Higgs et al., 2010) and putative Osedax
borings (Kiel et al., 2010). Despite these efforts, there has
been no detailed documentation of the morphology of indi-
vidual borings made by any Osedax species to date. The
results presented here provide the first quantitative docu-
mentation of individual Osedax borings. These detailed data
on the volume and surface area of the borings (Fig. 5)
provide an effective way to analyze the part of the Osedax
worm that is housed inside the bone. The wide range in the
size of borings measured allows us to address the variability
314 N. D. HIGGS ET AL.
in Osedax mucofloris boring and understand the processes
that influence it.
The morphology and development of Osedax mucofloris
borings
The individual Osedax mucofloris borings found in the
minke whale radius are consistently hemi-ellipsoidal (i.e.,
bowl-shaped) over a range of sizes (Figs. 4E, 5A), but they
are markedly different from borings found in the vertebral
processes (Figs. 6, 7). The consistency of morphology
within bones and the disparity between bones shows that the
shape of O. mucofloris borings changes depending upon the
type of bone inhabited. This plasticity in boring morphology
is also mirrored by the root tissues of O. mucofloris, which
appear more dendritic in individuals dissected from spongy
or degraded bone (e.g., Fig. 8B), and more globular in those
from denser bone (Fig. 4; C. Verna, Max Planck Institute of
Marine Microbiology; pers. comm.). Variation in root mor-
phology with bone type has also been noted anecdotally in
other Osedax species (Kiel et al., 2010; F. Pradillon, IF-
REMER; pers. comm.).
The root tissues of O. mucofloris appear to fill the entire
borings found in whale bones (Fig. 3), indicating that the
reconstructed borings are accurate representations of the
root morphology. The root morphology has been used as a
taxonomic trait in describing and comparing Osedax spe-
cies; however, in some species this may change once ex-
tracted from the bone (e.g., Osedax roseus Rouse et al.,
2008), partly because of the difficulty in dissecting the
entangled root tissue. Similarly, the ovisac and roots of
Osedax frankpressi Rouse et al., 2004, are inflated with a
clear fluid when in situ, but this is also lost during extraction
(Rouse et al., 2004). These results show that micro com-
puted tomography provides an accurate method of analyz-
ing and comparing the root morphology for multiple indi-
viduals in situ, although tendrils that are smaller than intra-
trabecular spaces may not be detectable without staining.
On the basis of the evidence presented here, we suggest
a model of O. mucofloris root growth that is mediated by the
structure of cetacean bone. Whale bones have an extremely
reduced cortical bone layer that encases the spongy trabec-
ular bone. This spongy bone is densest near the surface, then
decreases toward the center of the bone where marrow-rich
pore spaces reach diameters of 35 mm (see Felts and
Spurrell, 1965, 1966, for excellent images of this gradient).
The rate of boring appears to decrease with decreasing
density of the bone, leading to a narrowing of the borings
with depth and giving rise to their hemi-ellipsoidal mor-
phology. In denser bone, the rate of vertical root expansion
is proportional to lateral expansion (Fig. 5A). Where the
bone-density gradient changes rapidly over a short distance,
such as in the vertebral processes (Fig. 6B), the rate of
lateral root expansion into the densest bone outstrips verti-
cal expansion, producing the wide, shallow borings. Lateral
expansion continues to the point where individual borings
merge to form large subsurface cavities (Higgs et al., 2010)
and may lead to a competition between individuals in
Osedax populations. It is interesting that although the bone
is eroded, a thin layer covering the root tissues is always
maintained. It is not known how the worms differentiate this
bone from the bone that they erode, but it may correspond
to the thin cortical layer and obviously provides protection
to the reproductive organs and root tissues.
The erosion of the trabeculae (Fig. 4F) lends support to
the hypothesis that O. mucofloris is utilizing the bone tissue
itself, not the lipids held within the bone (Higgs et al.,
2010). This is the case for Osedax frankpressi and two other
Osedax species, which seem to rely on the collagen part of
the bone (Goffredi et al., 2005) for nutrition. The O. mu-
cofloris roots appear to have avoided the intratrebecular
spaces that contain lipid-rich bone marrow (Fig. 4F), and so
it is assumed that they are also utilizing the collagen fraction
of the bone for nutrition. It is unclear if this preference
applies to the entire genus, since Osedax japonicus Fujikura
et al., 2006, has been reported to live on spermaceti, a waxy
whale tissue (Fujikura et al., 2006: fig. 4E). In either case
the root tissues of Osedax seem to be well adapted to the
uptake of dissolved compounds as well as secretion of
enzymes (Katz et al., 2010), but more work is required to
fully understand nutrition in this genus.
The process of boring in Osedax mucofloris
The positive staining of Osedax mucofloris mucus indi-
cates the presence of AMPs (Fig. 8C), providing insight into
how this animal (lacking any hard tissues) may be able to
bore into bone. Together with the trabecular rinds (Fig. 4F),
this new evidence suggests that the boring process is chem-
ically mediated, since mechanical erosion would destroy
such fine structures. Additionally, the presence of col-
lagenolytic enzymes has previously been detected in
Osedax roots and attributed to their symbionts (Goffredi et
al., 2007; but see Katz et al., 2010). However, proteolytic
enzymes alone are almost certainly insufficient for the deg-
radation of bone tissue; an acid is needed to first degrade the
mineral fraction, since the collagen and mineral form a
mutually protective structure (Child, 1995). Once mineral
crystals have been dissolved, the enzymes can break down
the collagen, providing a potential food source. In the anal-
ogous process of vertebrate bone resorption, which has only
recently been elucidated, the bone is first demineralized at
pH 4.5, then the collagen matrix is broken down by en-
zymes (Teitelbaum, 2007). The AMPs may thus provide the
necessary conditions for the first stage of bone degradation
(mineral dissolution) by acting as chelating agents (Simkiss
and Tyler, 1957).
The erosion of bone by Osedax appears to proceed with
 OSEDAX BORINGS IN WHALE BONES 315

the roots penetrating the trabecular structures of the bone floris borings lack. The general concordance of these fossil
and chemically eroding the bone tissue within these trabec- borings with those known to be of Osedax origin (Figs. 2
ular struts (Fig. 4F). Higgs et al. (2010) suggested that and 4) substantiates their supposed origin. It also suggests
Osedax erosion progressed without regard to bone structure, that there may be interspecific, as well as intraspecific (this
but Figure 4F clearly shows that the fine-scale trabecular study), variability in boring morphology—a possibility that
structure of the bone influences the erosion process. This is currently being investigated.
micro-scale structural influence explains the macro-scale
morphology of the borings. Since the roots are guided by the The effects of Osedax borings on whale-fall communities
trabecular structure of the bone, they will proceed where
Osedax worms have been described as “foundation spe-
there are more trabeculae—that is, in the densest bone. As
cies” in whale-fall communities owing to their perceived
the trabeculae become less dense toward the interior of the
influence on the degradation of the whale skeletons (Braby
bone, fewer root projections will be growing in that direc-
et al., 2007). The maximum rate of bone erosion measured
tion. The hollow rinds of eroded trabecular bone have also
in this study (⬃6% per year) is greater than the 1% per year
been observed in fossil whale bone, where the erosion was
reported by Higgs et al. (2010), which can be attributed to
thought to be caused by microorganisms (Amano et al.,
the increased proportion of the radius that was exposed and
2007). It may be that erosion of bone by both Osedax worms
colonized by O. mucofloris. Both values appear lower than
and microorganisms is affected by the microstructural prop-
would be predicted from the observations of Lundsten et al.
erties of the bone.
(2010), who reported the destruction of an entire whale
The microborings found on the root-associated bone frag-
skeleton in about a decade. However, many of the Osedax
ments may be related to the dense bacterial community
species reported from these whale-falls are much larger than
(including Osedax symbionts) that has been observed in
O. mucofloris and often occur at higher densities (e.g.,
association with the mucous layer surrounding the root of
Rouse et al., 2008). Furthermore, the values reported here
Osedax mucofloris (Verna et al., 2010), and so aid Osedax
are based on bones that were exposed for 8 –12 months, and
boring. Alternatively, microorganisms unrelated to Osedax
the rate of bone degradation may increase after the loss of
activity may have produced the etchings shown here. The
cortical bone, as in the oldest vertebrae (VP2). It is likely
fact that the microborings are restricted to the surface of the
that other bioeroding organisms such as bone-eating gastro-
relatively fresh radius (Fig. 4F) suggests that they are
pods (Johnson et al., 2010), tanner crabs (Braby et al.,
caused by microorganisms in the seawater. There would be
2007), and boring microorganisms (Higgs et al., 2011) also
little advantage in the Osedax worm destroying the thin
play a significant role in bone degradation.
protective bone covering its roots.
The black staining around the bore holes in the minke
whale radius is indicative of iron sulfides that may have
Osedax in the fossil record precipitated when hydrogen sulfide leaking from the bones
came in contact with oxygenated seawater. This interpreta-
Documentation of modern Osedax borings provides an ac-
tion is further supported by the concurrent concentrations of
curate method for identifying the activity of these animals in
white filamentous bacterial mats associated with these “sul-
the fossil record. Since Osedax worms actively degrade the
fide halos.” This is the first evidence that Osedax borings
bones that they live on, it might be expected that their borings
may act as conduits of hydrogen sulfide from the interior of
are unlikely to be preserved as fossil traces. On the contrary,
the bone where lipids are being degraded, as hypothesized
the high sedimentation rates at the site we examined (and
by Treude et al. (2009). A consequence of this process is
personal observations of buried bones) suggest that it is quite
that the degree of Osedax boring on a whale skeleton would
possible for bones to become buried before they are completely
be related to the rate of sulfide release from the bones. This
destroyed, thus allowing their traces to be preserved.
may help explain why whale-fall communities that are
The morphology of Osedax mucofloris borings is mark-
dominated by Osedax appear to have a shortened sulfophilic
edly different from that of the fossil borings presented by
stage in comparison to those where Osedax are less preva-
Muñiz et al. (2010). Published pictures of Osedax roots in
lent (Smith, 2006; Braby et al., 2007).
situ show the tendency for the roots to be globular or
laterally expansive, and never tubular (Rouse et al., 2004,
Acknowledgments
2008; Kiel et al., 2010). The interpretation by Muñiz et al.
(2010) that the trace fossil Trypanites ionasi was caused by We are grateful for the expert assistance provided by Tomas
an Osedax-like animal is not supported by our evidence. In Lundälv in ROV operations and Richie Abel in micro-CT
contrast, the borings in dense bone presented by Kiel et al. scanning. NDH was supported by a CASE studentship (NE/
(2010) as fossil “Osedax borings” are shallow and globular G523755/1) from the Natural Environment Research Council,
like those of O. mucofloris, though they appear to be more UK, and TGD was supported by the Swedish Research Coun-
spherical and have a defined trunk section, which O. muco- cil. AGG is supported by a grant from the SynTax initiative. A
316 N. D. HIGGS ET AL.
part of this work was also supported by a European Commu-
nity ASSEMBLE grant, agreement no. 227799. The manuscript
was improved by the stimulating and helpful comments of two
anonymous reviewers.
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