See

discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/315660248

The anatomy of the blood vascular system of

the giant vestimentiferan tubeworm Riftia
pachyptila (Siboglinidae, Annelida): ....

Article in Journal of Morphology · March 2017

DOI: 10.1002/jmor.20677

CITATIONS READS

0 68

3 authors:

Nadya Rimskaya-Korsakova S. V. Galkin

Lomonosov Moscow State University P.P. Shirshov Institute of Oceanology
10 PUBLICATIONS 12 CITATIONS 90 PUBLICATIONS 583 CITATIONS

SEE PROFILE SEE PROFILE

Vladimir Malakhov
Lomonosov Moscow State University
239 PUBLICATIONS 1,115 CITATIONS

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Non-vent fauna of the Mid-Atlantic Ridge View project

Workshop “Comparative anatomy of invertebrates” View project

All content following this page was uploaded by Vladimir Malakhov on 13 September 2017.

The user has requested enhancement of the downloaded file.

Received: 17 August 2016 | Revised: 28 February 2017 | Accepted: 8 March 2017

DOI: 10.1002/jmor.20677

RESEARCH ARTICLE

The anatomy of the blood vascular system of the giant

vestimentiferan tubeworm Riftia pachyptila
(Siboglinidae, Annelida)

Nadezhda N. Rimskaya-Korsakova1 | Sergey V. Galkin2 | Vladimir V. Malakhov3

1
Department of Invertebrate Zoology,
Lomonosov Moscow State University, 1-12,
Abstract
Leninskie Gory, Moscow, 119234, Russia The giant dimensions of vestimentiferan Riftia pachyptila (Jones, 1981) are achieved thanks to the
2
Laboratory of Bottom Ocean Fauna, well-developed vascular system. In the vestimentum, there is a complicated net of lacunae, includ-
Shirshov Institute of Oceanology (RAS), 36, ing the brain blood supply and the ventral lacuna underlying the ciliary field. The trunk region has
Nakhimovski prospect, Moscow, 117997,
an extensive network of blood vessels feeding the gonads («rete mirabile»). The thick muscular lin-
Russia
3
ing of the mesenterial vessels in the trunk and the dorsal vessel in the opisthosome serves as an
Department of Invertebrate Zoology,
Lomonosov Moscow State University, 1-12, additional pump, pushing blood into numerous vessels in the segments. It was hypothesized that
Leninskie Gory, 119234, Moscow, Russia the blood envelope of the ventral blood vessel in the trunk provides the blood supply to the troph-
and the Far Eastern Federal University, osome. The 3D reconstruction has revealed that there are two vascular systems of the tentacular
Sukhanova, 8, Vladivostok, 690950, Russia
crown of R. pachyptila. Blood runs into the tentacles via axial afferent vessels, as described earlier
Correspondence
only for Riftia, and also via basal ones, as described for other vestimentiferans except Riftia. The
Nadezhda Rimskaya-Korsakova, Lomonosov
Moscow State University, Department of basal ones are poorly developed, and the number of lamellar blood vessels is small, indicating a
Invertebrate Zoology, Vorobievy Gory 1-12, lack of demand for these within huge R. pachyptila. It appears that the presence of these vessels
Moscow, Russian Federation. is the preserved ancestral state of Vestimentifera. In different portions of the dorsal vessel, the
Email: nadezdarkorsakova@gmail.com/
morphology of the intravasal body varies, depending on function.
rimskaya-korsakova@mail.bio.msu.ru

KEYWORDS
circulatory system, dorsal blood vessel, intravasal body, sinus valvatus, ventral blood vessel, vessels
of tentacles, vessels of trophosome

1 | INTRODUCTION between symbionts in the trophosome, and afterward distributes

The discovery of Vestimentifera was one of the most spectacular
events within zoology of the second half of the 20th century (Arp &
Childress, 1983; Bosch & Grasse, 1984; Cavanaugh, Gardiner, Jones,
Jannasch, & Waterbury, 1981; Felbeck, 1981; Jones, 1980, 1985;
Mane-Garzon & Montero, 1985; Van der Land & Nørrevang, 1975,
1977; Webb, 1969). Attention was particularly drawn to vestimenti-
feran tubeworms after the East Pacific hydrothermal vent communities,
in which tubeworms play an essential role, were found, although many
vestimentiferan species inhabit cold seeps. All adult vestimentiferans
lack a digestive system, and obtain their nutrients from chemoendo-
symbiontic bacteria cultured in trophosome tissue. A highly developed
circulatory system greatly contributes to the logistics of nutrition.
Firstly, the blood vessel network in the tentacles provides an intensive
exchange of gases between ambient seawater and the blood capillaries;
following this, the blood system delivers and spreads substances
organic matter to the tissues and organs of the host tubeworms
(Andersen, Jolivet, Claudinot, & Lallier, 2002; Bright & Sorgo, 2003;
Gardiner & Jones, 1993; Karaseva, Malakhov, & Galkin, 2012;

, 2004;
Malakhov, Popelyaev, & Galkin, 1996; Minic & Herve
Southward, Schulze, & Gardiner, 2005).
The unique organization of vestimentiferan tubeworms initially
caused researchers to classify them in a separate phylum, Vestimenti-
fera or Mesoneurophora (Jones, 1985; Mane-Garzon & Montero,
1985), although some authors, such as Van der Land and Nørrevang
(1977), were unconvinced, considering them to be a group within
Annelida. Today, Vestimentifera has undergone a demotion, and is
included in the family Siboglinidae (Rouse, 2001). Within Siboglinidae,
Vestimentifera is nested together with Sclerolinum with high support
€m, Smith, & Dahlgren, 2005; Glover
(Rouse, 2001; Glover, Källstro
et al., 2013; Eichinger, Hourdez, & Bright, 2013; Li et al., 2015, 2016;
Rouse, Wilson, Worsaae, & Vrijenhoek, 2015). According to a recent

Journal of Morphology. 2017;1–18. wileyonlinelibrary.com/journal/jmor V

C 2017 Wiley Periodicals, Inc. | 1
2 | RIMSKAYA-KORSAKOVA ET AL.

T A B LE 1 The studied specimens collected during cruises of the RV Akademik Mstislav Keldysh (AMK).

Length of Coordinates
Specimen# specimen (mm) Sex of the collection site Depth (m) R/V – station#, ROV, year

1 16 Female 27802,45’ N, 111822,80’ W, 1990 AMK-1519, Pisces-VII, 1986

Guaymas Basin

2 34 Female 09850,52’ N, 104817,52’ W, 2524 ffR - 4623, MIR-1, 2003

“98N EPR”

3 101 Female 27800,47’ N, 111824,57’ W 2001 ffR-4714, MIR-2, 2003

Guaymas Basin

4 308 Female 20819,86’ N, 109806,20’ W 2997 ffR-4679, MIR-1, 2003

“218N EPR”

5 79 Male 09850,52’ N, 104817,52’ W 2524 ffR-4623, MIR-1, 2003

“98N EPR”

synopsis (Karaseva, Rimskaya-Korsakova, Galkin, & Malakhov, 2016),
Vestimentifera comprises 19 species distributed between three groups
(infra-families), Lamellibrachiinae (Webb, 1969), Escarpiinae (Jones,
1985), and Tevniinae (Jones, 1985). Lamellibrachiinae and Escarpiinae
inhabit soft and rocky substrates in the areas of cold seeps and on the
periphery of hydrothermal vents, and are recorded everywhere except
areas in the Arctic and Antarctic; however, Tevniinae is known to
exclusively inhabit the rocky substrates of the Pacific Ocean hydrother-
mal vents (Karaseva et al., 2016; Schulze & Halanych, 2002).
The circulatory system of the vestimentiferan Lamelibrachia luymesi
has been described in detail by Van der Land and Nørrevang (1975,
1977). The arrangement of the circulatory systems of the medium-
sized tubeworm Ridgeia piscesae and the small-sized tubeworm Oasisia
alvinae are similar to those of Lamellibrachia (Karaseva et al., 2012;
Malakhov et al., 1996). In spite of the giant size of tubeworm Riftia
pachyptila, we know few details of the circulatory system of the animal.
There are the blood lacunae underlying the ventral ciliary field in the
vestimentum, the blood envelope of the ventral blood vessel in the
trunk, as well as the main afferent and efferent vessels of the tentacu-
lar apparatus are parallel to the body’s axis in Riftia, whereas in all other
vestimentiferan tubeworms these vessels are located in the transverse
plane (Gardiner & Jones, 1993; Jones, 1980, 1981, 1985). The latter
feature has been used by Jones (1985) as a basis for the taxonomic
division of Vestimentifera (which he regarded as a phylum) into two
classes: Axonobranchia (with a single species, R. pachyptila) and Basi-
branchia (with the rest of the species of tubeworms described so far).
Neither molecular data (Black et al., 1997; Halanych, Feldman, &
Vrijenhoek, 2001; Kojima et al., 2003), nor the organization of the
tentacle crown (Rimskaya-Korsakova & Malakhov, 2010; Rimskaya-
Korsakova, Malakhov, & Galkin, 2011) in R. pachyptila support the
classification by Jones (1985). Unfortunately, the latter studies have
not taken advantage of 3D reconstruction methods.
Moreover, some special organs (an intravasal body and a sinus val-
vatus) of the vestimentiferan circulatory system raise questions. The
intravasal body, or heart-body, or intravasal tissue (Gardiner, 1992), is a
strand of tissue attached to the ventral surface of the dorsal vessel in
the vestimentum and trunk have been described by Gardiner and Jones
(1993) as a flapper-valve structure preventing back flow from the
dorsal vessel into the mesenterial blood vessel, whereas Schulze (2002)
discussed its function as a hemopoetic organ. The sinus valvatus,
another vascular organ of unclear organization and function, is a valve-
like structure that seems to prevent backflow of blood into the ten-
tacles (Southward et al., 2005).
In the present article, we aim to describe the adult circulatory
system and its special organs (such as the intravasal body and sinus
valvatus) of R. pachyptila, using visualization with light microscopy,
histology and 3D modeling. A further aim of this work is to discover
whether there are morphological features of the vascular system of
R. pachyptila in comparison with other, much smaller, representatives
of the vestimentiferan tubeworms, such as R. piscesae and O. alvinae,
which might serve as adaptations to the huge size of the tubeworm.
2 | MATERIAL AND METHODS
2.1 | Collection and fixation
Specimens of Riftia pachyptila (Jones, 1980, 1981) were collected at
different latitudes of the East Pacific Rise (EPR) and in the Guaymas
Basin, Gulf of California, by the PISCES manned submersible during the
12th cruise of RV Akademik Mstislav Keldysh in 1986 and by MIR-1
and MIR-2 manned submersibles during its 22nd and 49th cruises in
1990 and 2003, respectively. Lengths of examined specimens and the
collection sites are listed in Table 1.
2.1.1 | Histology and LM photography
Four animals used for anatomical analysis were fixed in Bouin’s solution
and stored in 70% ethanol. The material was processed by the standard
histological procedure, including dehydration in an alcohol series and
embedding in paraffin, paraplast, or histowax. Transverse sections
(5 and 7 lm) were produced with a Leica RM 2125 microtome (Leica
Microsystems, Wetzlar, Germany), stained with Caracci hematoxylin,
and examined under a Zeiss Axioplan2 microscope equipped with
AxioCam HRm camera (Carl Zeiss Microscopy, LLC, USA) as well as
Leica DM5000 B equipped with Leica DFC425 C camera. Microscopic
images were optimized for contrast and level in Adobe Photoshop 7.0
(Adobe Systems, San Jose, CA, USA). Drawings were performed with
RIMSKAYA-KORSAKOVA ET AL.
Adobe Illustrator CS4. For visualization of the blood vessels in the ves-
timentum a specimen of 101 mm was pictured by Canon Power Shot
S90 camera.
2.1.2 | 3D modeling
Arrangements of vessels of the tentacular crown and the anterior-
most vestimentum were visualized with the software 3D-DOCTOR
3.5.040724 (Able Software Corporation of Lexington, USA), 3D vol-
umes of the sinus valvatus were made with the TrackEM plugin of
ImageJ/FIJI (Cardona et al., 2012). Alignment was performed in the
corresponding softwares by comparing the sections of adjacent
planes.
An image series of 61 transverse sections of the 34 mm specimen
was used for modeling of the circulatory system of the tentacular
crown. On each 2D section the brain, obturacular lobes, main tentacu-
lar blood vessels, basal tentacular vessels, axial tentacular vessels,
lamellar vessels, tentacular vessels, and obturacular vessels were
outlined.
An image series of 54 transverse sections of the 16 mm specimen
was used for modeling of the circulatory system of organs of the ante-
riomost vestimentum. On each 2D section the entire body, brain,
obturacular lobes, excretory tree and ducts, anterior perivascular coe-
lom, obturacular coelom, main efferent vessels, sinus valvatus were
outlined.
An image series of 16 transverse sections of the 16 mm specimen
was used for modeling of the sinus valvatus. On each 2D section the
main efferent tentacular vessels, the ventral vessel and the lateral
coelomic sacs of the sinus valvatus were outlined.
On the basis of the outlined boundaries three-dimensional (3D)
models were obtained. The smoothing tool used for natural perception
of the surface of objects. Interactive features as well as transparency
filter, different colors, and lighting effects applied to show complex and
hidden objects. Three-dimensional images under appropriate angles
were processed in Adobe Photoshop 7.0 (Adobe Systems, San Jose,
CA, USA).
3 | RESULTS
The vascular system of Riftia pachyptila includes the dorsal blood-
vessel-bearing heart, which pushes blood forward into the vessels of
the tentacles and obturacles, the ventral blood vessel that brings the
blood from the tentacular vessels and provides blood supply to the
organs of the vestimentum (the brain, the excretory tree, and the
musculature), the trunk (the trophosome and the reproductive system),
and the opisthosome (Figure 1A–C).
3.1 | Dorsal blood vessel
The dorsal vessel runs the length of the body (dv, Figure 1A–C). In the
vestimentum, it is in the cylindrical perivascular coelom, and is supported
by the dorsoventral mesentery (Figure 2A–D). The dorsal vessel wall
consists of scattered hemocytes, ECM layer and coelomic epithelium
(Figure 2A, B).
| 3
In the trunk, the dorsal vessel runs within the trunk coelom,
dorsally with respect to the trophosome (Figure 1A). The diameter of
the dorsal vessel in the trunk is ten times bigger than in vestimentum
(Figure 2A–C). The wall of the vessel is organized in the same way as in
the vestimentum. Sparse hemocytes adhere to the ECM layer.
3.2 | Intravasal body
In the dorsal vessel, there is an intravasal body (ib, Figure 1A–C) in the
posteriormost vestimentum and along the entire length of the trunk
(Figures 1A, B and 2B, C, E). It is located on the ventral side. The size
and shape of the intravasal body vary greatly in the sections made at
different levels.
The wall of the intravasal body consists of a layer of ECM and two
layers of cells: the external and internal layers (Figure 2B). The ECM
layer of the intravasal body is an extension of the ECM layer of the
dorsal vessel. The surface of the ECM facing the dorsal vessel lumen
bears scattered hemocytes. A well-developed epithelium is present on
the internal surface of the ECM of the intravasal body (Figure 2B–D).
On some sections, it looks like a single-layered epithelium, whereas on
others it forms parenchyma-like tissue, which almost entirely fills the
inner lumen of the intravasal body (compare Figure 2C and D). The
lumen of the intravasal body contains a flake-like material, with struc-
ture differing from the material of the dorsal vessel.
In the trunk, where the mesenterial vessel connects with the dorsal
vessel, the intravasal body bends like an arch over the place of connec-
tion, forming a valve-like structure (vib, Figure 2E). The wall structured
as in the intravasal body, but it has loose internal epithelium.
3.3 | Heart
This is a modified portion of the dorsal vessel, located in the anterior-
most vestimentum (h, Figures 1A and 3A, D). In most cases, the shape
of the heart lumen in fixed specimens is slit-like and flattened in the
dorsoventral direction. Inside of the ECM, there are hemocytes.
Outside of the ECM, there is a layer of cubic epithelial cells surrounded
by the longitudinal musculature and bundles of circular muscles. The
exterior-most layer of the heart wall is represented by circular muscula-
ture penetrated by the coelomic channels (pcc) that connect the
anterior and posterior perivascular coeloms. There is no clear border
separating the heart musculature from the musculature of the
vestimentum.
3.4 | Vessels of tentacular crown
Anterior to the heart, the dorsal vessel branches into four large vessels:
a pair of obturacular vessels and a pair of main afferent tentacular ves-
sels (obv, av, Figures 1A, 3B,C, and 4A, D). The perivascular coelomic
channels follow the branching vessels, and each vessel is surrounded
by the coelomic channels (obturacular and anterior perivascular,
respectively, obc, ac, Figure 5A–C). The main afferent tentacular vessels
are directed ventrolaterally, whereas obturacular vessels run ventrally
(Figure 3B).
4 | RIMSKAYA-KORSAKOVA ET AL.

F I G U R E 1 Scheme of the blood vascular system of vestimentiferan R. pachyptila. A. Blood supply in the vestimentum, obturacules and
tentacular crown. B. Blood supply in the trunk. C. Blood supply in the opisthosome. The dorsal vessel and afferent tentacular vessels shown
in light blue, the obturacular vessels shown in purple, the ventral vessels and efferent tentacular vessels shown in red, vessels connecting
the ventral and dorsal vessels shown in violet, the trophosome tissue shown in green, the body wall shown in grey. av – main afferent
tentacular vessels, aav – axial afferent tentacular vessels, aev – axial efferent tentacular vessels, alv – the most anterior lvv, atp – afferent
blood plexus of trophosome, atv1 – afferent vessels of trophosome arising from the bb, axv – axial trophosome vessels, bav – basal afferent
tentacular vessels, bb – bilobed bulges, bbv – brain blind vessels, bev – basal efferent tentacular vessels, cf – ventral ciliary field, cv –
vessels connecting mvv and lvv, dbv – dorsal brain vessel, dv – dorsal blood vessel, etv – efferent vessels of trophosome, ev – main efferent
tentacular vessels, gnc – genital coelom, h – heart, ib – intravasal body, lav – afferent lamellar vessels, lev – efferent lamellar vessels, lvt -
lateral vessels in trunk, lvv – lateral vestimental vessels, mbv – median brain vessel coming from dlv and penetrating ext and b, ms –
mesenterium, mv – mesenterial vessels, mvv – main vestimental vessels, ob – obturacules, obv – obturacular vessels, op – opisthosome, pcv
– small vessels coming from lvv and surrounding gl and muscles of vt, pv – peripheral vessels of the vw, pvv – paired vv in op, rv – radial
vessels of op, scv – semi-circular vessels of op, sv – sinus valvatus, t – trophosomal cords, tav – afferent tentacular vessles, tev – efferent
tentacular vessels, tr – trunk, vib – valve-like intravasal body preventing backflow from dv into mv, vl – ventral blood lacune, vlv – ventrolat-
eral brain vessels, vmv – ventral medial vessel, vof – capillaries feeding the maturing oocytes, vt – vestimentum, vv – ventral blood vessel,
vvb – blood envelope of vv in trunk, vvl – short vessels connecting vv and vl, vw – vestimental wings

A pair of obturacular vessels penetrates the brain, where they loop
in the form of a letter “U” and join the obturacular lobes (obv, Figures
1A and 4A–F). The obturacular vessels loop multiple times in a sinusoid
form, inside the cartilage tissue of the obturacles (Figure 4A, B).
The main afferent tentacular vessels run parallel to the dorsal side
of the brain (av, Figures 3B and 4D). Anterior to the brain, each vessel
divides into a basal afferent vessel (bav), directed ventrally and some-
what posteriorly, and an axial afferent vessel (aav) running parallel to
the body’s axis (Figures 1A and 4A, D). The basal afferent vessels give
off several semi-circular afferent lamellar vessels (lav) lying in the trans-
verse plane, and provide blood supply to the lamellae located at the
basal-most tentacular crown (Figures 1A and 4A, D). The numbers of
basal lamellae and lamellar vessels (lav, lev) depend on the size of the
specimen. For example, a specimen of 16 mm length has 5–7 pairs,
whereas a specimen of 34 mm length has 9; a larger specimen has a
higher number of lamellae and basal lamellar vessels. Each semi-circular
afferent lamellar vessel gives off afferent tentacular vessels (tav)
orientated parallel to the axis of the body (Figures 1A and 4A, D).
Axial vessels bring blood to the rest of the lamellae of the tentacu-
lar crown (Figures 1A, 4D–F and Figures 5–6 in Rimskaya-Korsakova
RIMSKAYA-KORSAKOVA ET AL. | 5

FIGURE 2 Dorsal vessel and intravasal body in the vestimentum and trunk of R. pachyptila. Microphotography (A–E, G) and drawings (F) of
transverse histological sections of the dorsal vessel of the juvenile of 16 mm in length (A–C, G) and female of 308 mm in length (D, E). A. The
dorsal vessel in the vestimentum. B. The intravasal body in the dorsal vessel in the posterior-most vestimentum. C. The dorsal vessel in the trunk.
D. The intravasal body of the dorsal vessel in the trunk. E. Valve-like intravasal body in the place of connection of the mesenterial and dorsal
vessels. F. Hypothetic scheme of the formation of the intravasal body. G. The dorsal vessel in the opisthosome. ce – coelomic epithelium, cmp –
circular muscles surrounding pc, ecm – extracellular matrix, d – dissepiments, dv – dorsal blood vessel, hc – hemocytes, ib – intravasal body, ie –
epithelium of the ib, ms – mesenterium, mv – mesenterial vessels, oc – coelom in the opisthosomal segments, pc – posterior perivascular coelom,
t – trophosomal cords, tc – trunk coelom, vib – valve-like intravasal body preventing backflow from dv into mv

et al., 2011). Each axial vessel gives off semi-circular lamellar vessels
(lav), which lie along the body axis, one by one (Figures 1A and 4A, D).
The lamellar vessels branch into many tentacular vessels (tav) orien-
tated parallel to the axis of the body. Lamellar vessels originating from
the axial vessels are not located in the same way as the ones from the
basal vessels. The latter lie in a transverse plane; that is, lamellar vessels
lie one by one, from the midline of the body to the periphery.
The system of efferent blood vessels of the tentacular crown
resembles the system of the afferent vessels (Figures 1A and 4C, E, F).
Efferent tentacular vessels (tev) bring blood into efferent lamellar
6 |
FIGURE 3 Structure of heart and dorsal vessel in the vestimentum
of R. pachyptila. Drawing (A) and microphotography (B–D) of
transverse histological sections at the anterior-most end of the
vestimentum. A. Heart region of the dorsal vessel. B–D. Sequence of
the sections showing the heart (D) branching into the afferent
tentacular and obturacular vessels (C, B). av – main afferent
tentacular vessels, b – brain, cmh – circular muscle bundles of the
heart wall, ecm – extracellular matrix, exd – excretory ducts, ext –
excretory tree, ev – main efferent tentacular vessels, h – heart, hc –
hemocytes, lm – longitudinal muscles, lmh – longitudinal muscle bun-
dles of the heart wall, mbv – median brain vessel coming from dlv and
penetrating ext and b, obc – obturacular coeloms, obv – obturacular
vessels, pcc – channels in the muscle wall of the heart, connecting ac
and pc, sdv – branching dorsal vessel into obv and av
vessels (lev); then to efferent axial and basal vessels (aev, bev); and then
to the paired main efferent vessels of the tentacular crown. The latter
run over the dorsal side of the brain; after the brain, they run ventrally
RIMSKAYA-KORSAKOVA ET AL.
and enter the sinus valvatus (sv). The histological peculiarities of the
vessels of the tentacular crown are described in Rimskaya-Korsakova
et al. (2011).
3.5 | Sinus valvatus
The organ lies posterior to the brain and ventrally in regard to the
excretory tree (sv, Figures 1A and 5A–C). A pair of lateral coelomic sacs
(lcs) is closely associated with the sinus valvatus (Figures 5D–F and 6A–
D). These two sacs constrict the lumen of the sinus valvatus and sepa-
rate the volume of the sinus into two portions: the dorsal and ventral
portions (Figures 5D–F and 6A–D). The dorsal portion is formed by the
fusion of the main efferent vessels, whereas the ventral portion gives
rise to the ventral vessel. Within the series of transverse sections, there
is no connection between these lateral sacs and the lumen of the sinus
valvatus. The inner epithelial lining and nuclei can be recognized in the
lateral sacs.
3.6 | Ventral blood vessel
This extends along the body from the sinus valvatus to the opistho-
some, where it is divided into two lateral vessels (sv, vv, pvv, Figure
1A–C). In the vestimentum, the ventral vessel runs inside the thickness
of the musculature, and in the trunk it lies along the trunk coelom
(Figures 7A and 8A). The shape of the vessel lumen in transverse
sections varies considerably along its length (Figure 7A, C).
In the vestimentum, the ventral vessel lacks specific musculature
(Figure 7A, C). In the trunk, the ventral vessel wall is complex, and light
microscopy is not sufficient to study all the details of this (Figures 8A,
B, D, 9A, and 10). Outside of the vessel, there is a blood envelope
(vvb). Distinct connections between the ventral vessel and the blood
envelope are seen on some sections. Between the walls of the ventral
vessel and the blood envelope there are narrow spaces, inside which
circular muscle bundles are visible on the inner and outer walls and the
large nuclei. The presence of the latter two structures provides evi-
dence for the consideration of these spaces as coelomic cavities. The
blood envelope is divided by vertical trabeculae connecting its outer
and inner walls.
3.7 | Ventral blood lacuna
This lies ventrally to the ventral vessel, and follows the shape of the
ventral ciliary field in the vestimentum (vl, Figures 1A and 7E, F). The
lacuna is irregularly connected, with venal vessels via short vessels (vvl),
and gives rise to branching blood vessels underlying the epidermis. The
latter are clearly visible from the outside, particularly at the posterior
margin of the vestimental fold (pv, Figures 1A and 7E).
3.8 | Blood supply to the brain
This is provided by the vessels of the anterior vestimentum (Figures 1A
and 7G). Along the ventrolateral surface of the brain, there are two
large, symmetrical ventrolateral brain vessels (vlv) which flow into the
ventral blood vessel posterior to the sinus valvatus. These vessels are
RIMSKAYA-KORSAKOVA ET AL. | 7

FIGURE 4 Vascular system of tentacular crown of vestimentiferan R. pachyptila. 3D-reconstructions of organization of the vascular system
in the tentacular crown (A, C, D–F), afferent tentacular and obturacular vessels shown in blue, efferent tentacular vessels shown in red,
obturacules shown in orange, brain shown in yellow, for clear visualization the number of tentacles twice decreased and the size of the
tentacular vessels (tav, tev) increased in five times, length of XYZ axes is 0.4 mm, transparency filter applied for demonstration of the
vessels inside obturacules and brain (C–E), for volume representation transverse sections at the distal end of the reconstructed fragment
shown (E, F); digital photography of a specimen (B). A. Posterior lateral view of the system of afferent vessels of tentacular crown and
obturacular vessels, note main afferent vessels (av) branching into the basal and axial afferent vessels (bav, aav). B. Dorsal view of
obturacular lobes showing the obturacular vessels. C. Posterior lateral view of the afferent and efferent vessels of tentacular crown, note
the basal and axial efferent vessels (bev, aev) coming into main efferent vessels (ev). D. Lateral view of afferent vessels of tentacular
crown, note main afferent vessels (av) branching into the basal and axial afferent vessels (bav, aav). E, F. Frontal view of afferent and
efferent tentacular vessels; transverse section at the base of tentacle crown (E) and at the level of brain (F). av – main afferent tentacular
vessels, aav – axial afferent tentacular vessels, aev – axial efferent tentacular vessels, b – brain, bav – basal afferent tentacular vessels, bbv
– brain blind vessels, bev – basal efferent tentacular vessels, ev – main efferent tentacular vessels, h – heart, lav – afferent lamellar vessels,
lev – efferent lamellar vessels, ob – obturacules, obv – obturacular vessels, tav – afferent tentacular vessles, te –free tentacles, tev – efferent
tentacular vessles, vt – vestimentum, vw – vestimental wings

formed from the fusion of the many small veins that make up the com- sels (alv) to the ventrolateral brain vessels (Figure 7G). The dorsal brain
plex network on the ventral and ventrolateral sides of the brain. Over vessels gather blood from the smaller vessels making up the dorsolat-
the dorsal side of the brain, there is one more pair of larger vessels— eral brain network. Some of the smaller vessels bind the dorsal brain
dorsal brain vessels (dbv)—which run from the anteriormost lateral ves- vessel and the ventrolateral brain vessels.
8 | RIMSKAYA-KORSAKOVA ET AL.

FIGURE 5 Structure of sinus valvatus and vessels in the anterior vestimentum of R. pachyptila. 3D models showing the position of the
sinus valvatus, afferent and efferent vessels, obturacular vessels and median brain vessel (A–C). Microphotography of histological sections’
sequence of the sinus valvatus from anterior end to posterior one (D–F), diagrams under photographs show the level of each section.
Microphotography of median brain vessel, mbv (G–H). A. 3D-model shows position of the main vessels in the anterior vestimentum. B–C.
Posterior and lateral views of 3D-models showing the vessels arrangement inside the tissues of the excretory tree and brain. D. Lateral coe-
lomic sacs (lb) make slit-like lumen of the ventral vessel. E. Main efferent vessels entering the sinus valvatus which lateral sacs narrow the
lumen of the ventral vessel lumen and prevent backflow from the ventral vessel to the efferent vessels, asterisks show the places of con-
nection of the mbv and sv. F. The ventral vessel and posterior-most end of lateral sacs of the sinus valvatus. G–H. Median brain vessel, mbv,
runs through the excretory tree as shown on C. ac – anterior perivascular coelom, b – brain, ce – coelomic epithelium, ev – main efferent
tentacular vessels, exd – excretory ducts, exs – excretory sacs, ext – excretory tree, g – remnants of gut, lcs – lateral coelomic sacs of sv,
mbv – median brain vessel coming from dlv and penetrating ext and b, nc – nerve cord, ob – obturacules, obc – obturacular coeloms, obv –
obturacular vessels, sv – sinus valvatus, vv – ventral blood vessel, vw – vestimental wings

The unpaired median brain vessel (mbv) arises from the dorsal por- ventrally penetrating the brain tissue (Figures 1A, 3B–D, and 5C, E, G,
tion of the sinus valvatus. It passes through the excretory tree, runs H). The median brain vessel connects with the paired dorsal brain ves-
over the saddle between the two lobes of the brain and then turns, sels through several transverse vessels (Figure 1A).
RIMSKAYA-KORSAKOVA ET AL. | 9

FIGURE 6 Vascular system arrangements in the vestimentum of R. pachyptila. 3D-models of organization of the sinus valvatus (A–C) and a
light microphotograph (D) of the series of transverse section based on which the structure of the sinus was visualized. The main efferent
vessels of tentacles and the ventral blood vessel are shown in pink, coelomic cavities are shown in green. A. Anterior and lateral view of
the sinus valvatus. B. Posterior and lateral view of the sinus valvatus. C. Posterior view of the coelomic cavities, it is seen the internal shape
of the cavities. Although internal walls of both coelomic sacs are very close to each other, but they are separated. D. Transverse section
through the mid portion of the sinus. ev – main efferent tentacular vessels, lcs – lateral coelomic sacs of sv, sv – sinus valvatus, vv – ventral
blood vessel

3.9 | Other peripheral vessels of the vestimentum (main
vestimental vessels and lateral vestimental vessels)
Paired main vestimental vessels (mvv) branch off from the main affer-
ent tentacular vessels in the anterior vestimentum (at the base of the
obturacular lobes), run symmetrically in the median plane of the vesti-
mentum and enter the ventral vessels in the posterior vestimentum
(Figures 1A and 7A, D).
Lateral vestimental vessels (lvv) run at the periphery of the vesti-
mentum and deliver blood to the musculature of the vestimental wings
(Figures 1A and 7A–D). These vessels are symmetrical: each lateral ves-
timental vessel forms a crescentic loop with inner and outer branches.
Inner branches (left and right) originate from the common root at the
dorsal side of the ventral vessel (Figures 1A and 7C), rise dorsally to
the vestimental wings, pass to the outer branches, return ventrally and
re-enter the ventral vessel. Both the inner and outer branches of the
lateral vestimental vessels give off numerous small vessels, which
branch into the musculature of the vestimentum. The inner branches
of the lateral vestimental vessels (lvv) are linked to the main vestimental
vessels (mvv) via short connecting vessels (cv) (Figures 1A and 7D).
The most anterior lateral vestimental vessels (alv) originate not from
the ventral blood vessel but from paired ventrolateral brain vessels (vlv),
and do not connect with the main vestimental vessels (Figure 1A). The
inner branches of the anterior lateral vestimental vessels connect to the
dorsal brain vessels involved in blood supply to the brain (see above).
3.10 | Trophosome blood supply
In the trunk region, the bilobed bulges (bb) form periodically on the
dorsal side of the ventral blood vessel (Figures 1B, 8E, 9A, and 10).
They are covered by the blood envelope, which is a continuation of the
ventral vessel blood envelope. The blood envelope of the bilobed
bulges gives rise to the short afferent trophosomal vessels (atv1). The
afferent trophosomal vessels sometimes originate from the blood
plexus surrounding the genital coeloms (Figure 9C). The trophosome (t)
of R. pachyptila consists of a series of longitudinal cylindrical cords.
Afferent vessels reach the closest trophosomal cords, and enter the
afferent trophosomal plexus, (atp) entwining the cords from the
outside (Figures 1B, 9A, E, and 10). The afferent plexus of
adjacent trophosomal cords is linked via secondary afferent vessels
10 | RIMSKAYA-KORSAKOVA ET AL.

F I G U R E 7 Vestimental vascular system of R. pachyptila. Light microphotographs of transverse sections (A–D, F, G) and digital photograph
of a specimen vetimentum (E). A. Overview of section of the middle part of the vestimentum. B. Blood supply in the vestimental wings. C.
Lateral vessels coming from the ventral vessel. D. Vestimental vessels connecting to lateral vessels. E. Ventral view of specimen showing
the ventral blood lacune, note peripheral vessels of the posteriormost margin of the vestimental wings. F. Ventral blood lacune underlying
the ciliary field. G. The blood supply of the brain. b – brain, cf – ventral ciliary field, bbv – brain blind vessels, cv – vessels connecting mvv
and lvv, dbv – dorsal brain vessel, dv – dorsal blood vessel, g – remnants of gut, gl – tupipar glands, lvv – lateral vestimental vessels, mvv –
main vestimental vessels, nc – nerve cord, obc – obturacular coeloms, obv – obturacular vessels, pc – posterior perivascular coelom, pcv –
small vessels coming from lvv and surrounding gl and muscles of vt, pv – peripheral vessels of the vw, vl – ventral blood lacune, vlv –
ventrolateral brain vessels, vv – ventral blood vessel, vw – vestimental wings

(atv2, Figures 9E and 10). The fine vessels of the afferent trophosomal
plexus give rise to capillaries, which penetrate the trophosome tissue
and reach the axial trophosome vessel (axv) extending along the central
part of each trophosome strand. From the axial trophosome vessels,
blood is supplied to the mesenterial vessel through the efferent troph-
osome vessels (etv, Figures 9D and 10).
3.11 | Mesenterial blood vessel
The mesenterial blood vessel (mv) runs along the trunk, ventrally to the
dorsal vessel (mv, Figure 9E). It is suspended by the dorsoventral mesen-
tery of the trunk coelom. It is the third largest vessel in the trunk after the
dorsal and ventral vessels. The wall of the mesenterial vessel is thick
(Figure 9D, F): outside of the folded ECM there are a few layers of circular
RIMSKAYA-KORSAKOVA ET AL. | 11

FIGURE 8 Trunk vascular system: structure of the ventral vessel wall, lateral vessel and blood supply of male reproduction system in the
trunk of R. pachyptila. Light microphotographs of transverse sections of vessels in the trunk (A–E). A. Overview of the ventral vessel of a
juvenile tubeworm. B. Fragment of the ventral vessel wall in close vicinity to the ventral mesenterium. C. Blood supply of the common and
feather-like longitudinal muscles. D. Pore providing connection of a lumen of the ventral vessel and blood envelop of the vessel. E. Blood
supply of the male germinal epithelium, contribution of the blood envelope of the ventral vessel. ce – coelomic epithelium, cmb – circular
muscle bundles, bb - bilobed bulges, ecm – extracellular matrix, ep – epidermis, es – spermatogonia, fm – feather-like muscles, ge – germinal
epithelium, gnc – genital coelom, gnd – gonoduct, hc – hemocytes, lm – longitudinal muscles, lvt - lateral vessels in trunk, ms – mesenterium,
t – trophosomal cords, tc – trunk coelom, tce – epithelium of trunk coelom, vp – opening connecting blood envelope and lumen of vv, vv –
ventral blood vessel, vvb – blood envelope of vv in trunk, vvc – coelomic envelope of vv

and longitudinal muscles. The mesenterial vessel periodically lifts dorsally,
and fuses with the dorsal vessel (Figures 1B, 2E, 9D, and 10). The valve-
like intravasal body (vib) appears at the places where the mesenterial ves-
sel joins the dorsal vessel (see above, Figures 2E and 9D).
3.12 | Blood supply to the reproductive system
The reproductive system (in both female and male) is supplied with
blood from the blood envelope of the ventral vessel (Figures 1B, 8E,
9A–C, E, G, and 10). In males, there is no specialized system of blood
vessels for the germinal epithelium in gonocoels. Genital coeloms are
closely adjacent to the dorsal wall of the ventral vessel (Figure 8E).
Their blood supply is provided by small blood lacunae arising from the
blood envelope of the ventral vessel wall.
In the female, there is a pair of short vessels (gnv) providing blood
supply to the germinal epithelia of each genital coelom and feeding the
maturing oogonia and oocytes (Figure 9A–C, G). The capillaries form a
complex blood capillaries network (“rete mirabile”), which can be
12 | RIMSKAYA-KORSAKOVA ET AL.

FIGURE 9 Mesenterial vessel, blood supply of the trophosome and female reproduction system of R. pachyptila. Light microphotographs
of transverse sections of the trunk of adult female specimen of 308 mm in length. A. Paired collateral vessels formed by the ventral vessel
wall, arrows show the directions of the vessels formation. B. Vessels of the germinal epithelium receiving blood from blood envelop of the
ventral vessel. C. Afferent vessels of the trophosome coming from the ventral vessel blood envelop. D. Efferent vessels of the trophosome
come into the mesenterial vessel entering the dorsal vessel, note the valve-like intravasal body preventing backflow from the dorsal vessel
into the mesenterial vessel. E. Overview of the trophosome blood supply. F. Structure of the mesenterial blood vessel. G. Capillary network
in the gonadal coelom («rete mirabile»). atp – afferent blood plexus of trophosome, atv1 – afferent vessels of trophosome arising from the
bb, atv2 – afferent vessels of trophosome linking the adjacent t, axv – axial trophosome vessels, bb – bilobed bulges, cmb – circular muscle
bundles, cmb – circular muscle bundles, dv – dorsal blood vessel, ecm – extracellular matrix, etv – efferent vessels of trophosome, ge – ger-
minal epithelium, gnc – genital coelom, gnd – gonoduct, gnv – vessels providing blood supply for germinal epithelium, lmb – longitudinal
muscle bundles, ls – lower level of capillary network of gnc («rete mirabile»), ms – mesenterium, mv – mesenterial vessels, t – trophosomal
cords, tc – trunk coelom, us – upper level of capillary network of gnc («rete mirabile»), vib – valve–like intravasal body preventing backflow
from dv into mv, vv – ventral blood vessel, vvb – blood envelope of vv in trunk
RIMSKAYA-KORSAKOVA ET AL. | 13

FIGURE 10 Scheme of blood supply of the trophosome of R. pachyptila. Blood shown in grey, arrows show the direction of blood flow. atp –
afferent blood plexus of trophosome, atv1 – afferent vessels of trophosome arising from the bb, atv2 – afferent vessels of trophosome linking the
adjacent t, axv – axial trophosome vessels, bb – bilobed bulges, dv – dorsal blood vessel, etv – efferent vessels of trophosome, gnc – genital coelom,
gnv – vessels providing blood supply for germinal epithelium, ib – intravasal body, mv – mesenterial vessel, t – trophosomal cords, tc – trunk coelom,
vof – capillaries feeding the maturing oocytes, vv – ventral blood vessel, vvb – blood envelope of vv in trunk, vvc – coelomic envelope of vv

divided into two levels: lower and upper (ls and us). The lower level of
capillaries (directly connected to the axial vessels) is a dense knot of
fan-like capillaries. At the posterior of the genital coeloms, there is an
upper level of the capillaries that is mostly oriented longitudinally. In
the series of histological sections, it can be seen that the capillaries of
the upper level are a continuation of the capillaries of the lower level.
The vast majority of oogonia and oocytes are adjacent to the walls of
the capillaries of the lower level.
Gonoducts are supplied with blood by the same short vessels (gnv)
that provide blood for the germinal epithelia of the genital coeloms
(Figure 9B, C).
3.13 | Other peripheral vessels of the trunk
Paired semi-circular lateral vessels (lvt) run at the periphery of the trunk
and deliver blood to the longitudinal musculature (common and
feather-like) as well as the tubipar glands (Figure 1B). These branch off
from the ventral median vessels (vmv), which in their turn arise regu-
larly from the blood envelope of the ventral vessel and run ventrally
within the ventral mesentery.
3.14 | Blood supply to the opisthosome
Both the dorsal and ventral vessels extend to the segmented opistho-
some (Figures 1C and 11A–D). In its first segments the ventral vessel
bifurcates into the paired ventral vessels (pvv) that are connected to
the dorsoventral mesentery. Semi-circular vessels (scv) originate from
the paired ventral vessels, run along the perimeter of the segment and
enter the dorsal vessel. Numerous radial vessels (rv) extend from the
semi-circular vessels and reach the dorsal vessel. Both semi-circular
vessels and radial vessels are supported by dissepiments of the opis-
thosomal segments. The diameter of the dorsal vessel gradually
decreases to the posterior, and in the final segments the lumen of
the vessel becomes indistinguishable. The thick layer of circular and
14 | RIMSKAYA-KORSAKOVA ET AL.

FIGURE 11 Blood supply of opisthosome of R. pachyptila. Light microphotographs of transverse sections (A–D). A. Radial vessels locating
in the opisthosomal dissipiments. B. Overview of the vessels of the opisthosome: the paired ventral vessel, dorsal vessel and semi-circular
vessels. C, D. Close-up of the semi-circular vessels (C, D) and the ventral vessel in the anterior part of the opisthosome (D). d – dissepi-
ments, dv – dorsal blood vessel, ep – epidermis, gl – tupipar glands, ms – mesenterium, nc – nerve cord, oc – coelom in the opisthosomal
segments, op – opisthosome, pvv – paired vv in op, scv – semi-circular vessels of op, rv – radial vessels of op, vv – ventral blood vessel

longitudinal muscle fibers is adjacent to the dorsal vessel wall (Figure Blood is pushed forward by the heart, that is, the muscle wall thicken-
11A), whereas the walls of the ventral, semi-circular and radial vessels ing of the dorsal vessel.
are formed from a thin and sparse layer of flattened epithelial cells, in The main difference between the circulatory systems of R. pachyp-
which the ECM layer is indistinguishable (Figure 11A, C, D). tila and other species of Vestimentifera lies in the anatomical organiza-
tion of the tentacular crown circulatory system. In most

4 | DISCUSSION vestimentiferan tubeworms, lamellar vessels are oriented transversally

and arranged concentrically (in accordance with the arrangement of
4.1 | Riftia and other vestimentiferan tubeworms in the lamellae). According to Jones (1985), in R. pachyptila there are axial

the light of the current phylogeny of the group
From the initial description of the circulatory system of Riftia pachyptila
(Jones, 1980, 1985), it can be seen to differ significantly from the orga-
nization of the circulatory system of other vestimentiferan tubeworms
for Lamellibrachia barhami (Webb, 1969), L. luymesi (Van der Land &
Nørrevang, 1975, 1977); Ridgeia piscesae (Malakhov et al., 1996), Oasi-
sia alvinae (Karaseva et al., 2012).
In general, R. pachyptila has a very well-developed circulatory sys-
tem of a closed type, which resembles the general organization of the
circulatory system of other siboglinids and annelids (Brusca & Brusca,
2003; Fransen, 1988; Gardiner & Jones, 1993; Gardiner, 1992; Ivanov,
1960; Malakhov et al., 1996; Pilger, 1993; Van der Land & Nørrevang,
1977), with two main vessels (dorsal and ventral) with blood flows in
opposite directions. Anteriorly, the dorsal and ventral vessels are con-
nected via the vascular system of the tentacular crown; posteriorly,
they are connected through the vascular system of the opisthosome.
vessels (afferent and efferent) from which lamellar vessels depart. The
latter are not arranged in transverse planes and concentrically, but
sequentially along the axis of the body. Based on the peculiarities of
the circulatory system, Jones (1985) divides Vestimentifera into two
classes: Axonobranchia (consisting solely of R. pachyptila) and Basibran-
chia (all other vestimentiferans).
This study shows that in the circulatory system of the R. pachyptila
tentacular crown the main paired afferent (as well as efferent) tentacu-
lar vessels (“dorsal vessels” in Jones, 1985) form two branches: the axial
branch (which divides anteriorly as described by Jones, see. Figure 1.1
in Jones, 1985) and the basal branch (which provides branches to sev-
eral lamellar vessels of the most basal lamellae). The arrangement of
the circulatory system of the tentacular apparatus of R. pachyptila
includes two systems that are characteristic of both Axonobranchia
and Basibranchia. The circulatory system of R. pachyptila displays ele-
ments of the organization of the other representatives of Vestimenti-
fera in the basal arrangement of lamellar vessels and the similar
RIMSKAYA-KORSAKOVA ET AL.
sequence of the basal lamellae origin (the oldest lamella being the clos-
est to the base of the basal tentacular vessel). The presence of the
basal lamellar vessels is a common feature in R. pachyptila and other
vestimentiferans. This is likely to be an ancestral feature of
Vestimentifera.
In Riftia, unlike the other Vestimentifera, there is a pair of the ves-
timental processes lying along the obturacular lobes and containing the
pairs of axial afferent and efferent tentacular vessels (Rimskaya-Korsa-
kova & Malakhov, 2010; Rimskaya-Korsakova et al., 2011). The axial
tentacular vessels due to the vestimental processes extend from the
base of tentacles to the foremost tentacular lamellae. As a lamella
departs from the vestimental process, a lamellar vessel is given off by
an axial tentacular vessel (both afferent and efferent). We assume that
the first lamellar vessel is given off from the axial vessel posteriorly,
and the last one anteriorly.
Basal tentacular vessels are located at the very base of the tentac-
ular crown, and do not lie inside the vestimental processes as in the
other vestimentiferans.
The present study of specimens of various sizes of R. pachyptila
from “98N EPR” and “218N EPR” confirms such an organization. Mala-
khov and Galkin (1998) noted that the development of an axial tentac-
ular vessel is associated with the large body size of R. pachyptila, which
requires more tentacles to provide oxygen and hydrogen sulphide. The
organization of the tentacular crown of Riftia provides the formation of
an unlimited number of tentacular lamellae via elongation of the ten-
tacular crown, without a significant increase in the diameter of the
body and the tube.
Another unusual feature of the circulatory system of R. pachyptila
and other vestimentiferans consists of the development of a vast lacu-
nar system in the vestimentum (i.e., the ventral lacunae underlying the
ventral ciliary field, or the numerous blind sinuses of the brain blood
supply) and an extensive network of blood vessels feeding the gonads
(in females). This is a particularly important peculiarity of R. pachyptila,
and arises from the large sizes of the internal organs requiring blood
supply. In addition, the sophisticated structure of the ventral vessel in
the trunk provides an efficient blood supply in the trophosome of R.
pachyptila. It is possible that the additional blood envelope, with strong,
muscular walls, allows for an increase in pressure to push the blood
through the numerous trophosomal vessels.
It is notable that the blood supply in the trophosome of L. luymesi
(Van der Land & Nørrevang, 1977) is complicated by the presence of a
pair of collateral vessels (vasa trophosomales collaterales) running paral-
lel to the ventral vessel. It is probable that these vessels in L. luymesi
are needed for the same reason as a blood envelope in Riftia. In Ridgeia
(Malakhov et al., 1996) and Oasisia (Karaseva et al., 2012) there are nei-
ther collateral vessels nor a blood envelope of the ventral vessel.
In all vestimentiferans investigated so far, the heart is located in
the same position, on the dorsal vessel in the anterior part of the vesti-
mental region (Gardiner & Jones, 1993; Karaseva et al., 2012; Mala-
khov et al., 1996; Southward et al., 2005; Van der Land & Nørrevang,
1977; Webb, 1969). The position of the heart on the dorsal vessel in
Vestimentifera is common throughout Annelida, although several
| 15
annelids possess additional hearts that vary in position and number
considerably (Brusca & Brusca, 2003; Gardiner, 1992). We suggest
here that in Vestimentifera, the musculature of the heart is derived
from the dramatically developed musculature of the vestimentum,
since there are perivascular coelomic channels penetrating the
muscular wall of the heart (Van der Land & Nørrevang, 1977; Malakhov
et al., 1996; and our data on R.pachyptila). So far, this feature has not
been found in other annelids.
In Riftia, due to the huge size, there are some additional heart-like
structures, that is, parts of different vessels with a strongly developed
muscular wall. In the opisthosome, the dorsal vessel has a significant
muscular wall, and may possibly work as a posterior pump or a kind of
additional opisthosomal heart. Considerable thickening of the muscle
layer has been revealed in the main afferent vessels of tentacles, and in
the mesenterial vessel in the areas where it connects to the dorsal
vessel.
4.2 | The intravasal body
The intravasal body in the dorsal vessel lumen has been documented in
all vestimentiferans exposed to histological surveys so far (Andersen
et al., 2004; Gardiner & Jones, 1993; Karaseva et al., 2012; Malakhov
et al., 1996; Schulze, 2002; Southward et al., 2005; Van der Land &
Nørrevang, 1977). In R. pachyptila, as well as in R. piscesae (Malakhov
et al., 1996), the structure of the intravasal body varies in different
parts of the dorsal vessel; it is either a cylinder of small diameter, or a
loose tissue process occupying the majority of the lumen. According to
Schulze (2002), Escarpia laminata, Arcovestia ivanovi, and Seepiophila jonesi
have the intravasal body as a compact cell cluster of similar structure
throughout the dorsal vessel.
Several hypotheses on the nature of the intravasal body have
been put forward. Van der Land and Nørrevang (1977) considered the
intravasal body in L. luymesi to be a proliferation of endothelial tissue,
although the authors noted that the intravasal body consisted of cells
with round nuclei and a large amount of cytoplasm, which differed con-
siderably from the endothelial tissue. Ivanov (1960), who found the
intravasal body in frenulates, also thought that it was a derivative of
the endothelium.
In R. pachyptila, the intravasal body wall consists of the ECM layer,
scattered hemocytes, the dorsal vessel on the surface of the ECM fac-
ing the lumen, and a well-developed epithelium on the surface of ECM
facing the inner lumen of the intravasal body. A similar structure of the
intravasal body was described in Ridgeia at the ultrastructural level
(Schulze, 2002). Based on this structure, it is logical to assume that the
intravasal body is an invagination of the wall of the dorsal vessel (Fig-
ure 2F). Based on this assumption, the inner epithelium of the intra-
vasal body appears to be a coelomic lining, and the inner lumen of the
intravasal body is a nothing, but a coelom. This assumption requires
verification by studies on early juveniles of Vestimentifera.
Homologous structure to the vestimentiferan intravasal body
might be a heart body found in some polychaete annelids, like Cossuri-
dae (Zhadan, Vortsepneva, & Tzetlin, 2015), Terebellidae (Kennedy &
Phillips Dales, 1958), Flabelligeridae (Spies, 1973) and Alvinellidae
16 | RIMSKAYA-KORSAKOVA
(Jouin-Toulmond, Augustin, Desbruyeres, & Toulmond, 1996; Schulze,
2002). It consists of a single layer of pseudostratified columnar epithe-
lial cells, which represent involuted peritonium (Gardiner, 1992), like in
vestimentiferan intravasal body. The heart body is placed within a poly-
chaete heart region of the dorsal vessel, whereas the intravasal body is
extended along the dorsal vessel of the vestimentiferan trunk and
vestimentum.
Functions of the vestimentiferan intravasal body are debated. The
polychaete heart body is assumed to perform the function of blood for-
mation (Kennedy & Phillips Dales, 1958; Meyer, 1887), later the hema-
topoietic function is considered to be proven (Braunbeck & Dales,
1985; Dales & Pell, 1970; Friedman & Weiss, 1980; Mangum & Dales,
1965; Potswald, 1981). Based on the structural similarity to heart
bodies, the same function of the intravasal bodies was assumed
(Schulze, 2002). Although, Gardiner and Jones (1993) suggested that
the intravasal body could function as a valve to prevent the backflow
of blood from the dorsal vessel to the mesenteric. This seems very
likely for the areas in which the mesenterial vessel connects with the
dorsal one.
4.3 | Sinus valvatus
The sinus valvatus is the most enigmatic organ of the circulatory system
of vestimentiferan tubeworms. Van der Land and Nørrevang (1977)
was the first to reconstruct this organ and to propose a diagram of the
internal anatomy (see Figure 11 in cited work). All recent authors
regard the sinus structure in the same way (Gardiner & Jones, 1993;
Karaseva et al., 2012; Malakhov et al., 1996; Southward et al., 2005).
The sinus valvatus is the widest anterior part of the ventral vessel into
which the main efferent tentacular vessels flow. At the same time, the
blood passes through the narrow slit between the two lateral sacs into
the expanded ventral part of the sinus, from which the ventral vessel
originates. All authors studying the structure of Vestimentifera have
interpreted the lateral sacs as derivatives of the sinus. On the diagrams
(see Figure 11 in Van der Land and Nørrevang, 1977; see Figure 9ffl in
Southward et al., 2005), the slit-like holes are depicted as connecting
the widest ventral part and lateral sacs of the sinus. Our careful exami-
nation of the series of the sections of the sinus valvatus of R. pachyptila
provides no evidence that such holes exist. Moreover, the internal epi-
thelial lining of the lateral sacs is seen on photographs of Riftia (Figure
5D–F) and Oasisia (see Figure 6a in Karaseva et al., 2012). Based on
the foregoing, we hypothesize that the lateral sacs of the sinus valvatus
are coelomic cavities; this offers an explanation for the presence of the
explicit continuous epithelial lining of the sacs, and for the absence of
connection between their lumens and the lumen of the sinus. If one
accepts this hypothesis, it would be easy to understand the functioning
of the organ. The lateral coelomic sacs attach closely to each other and
almost occlude the lumen of the sinus (Figures 5D–F and 6A–D). Blood
passes the narrow slit between the sacs and enters the ventral vessel.
It is therefore possible that the lateral coelomic sacs of the sinus valva-
tus prevent backflow from the ventral vessel to the efferent vessels of
the tentacular crown. The wall of the sinus valvatus lacks muscles, and
it is possible that the direction of the blood flow is triggered by the
ET AL.
musculature of the efferent vessels and the massive musculature of
the vestimentum. To verify our hypothesis, we have already made
study of the sinus valvatus with transmission electron microscopy of
another vestimentiferan species Ridgeia piscesae (the unpublished data
are not provided in this study).
4.4 | Synopsis
This study of the vascular system of the largest vestimentiferan tube-
worm Riftia pachyptila highlighted unique morphological traits of the
species distinguishing it from other smaller vestimentiferans. R. pachyp-
tila develops a complicated net of lacunae of the brain blood supply
and ventral lacuna underlying the ciliary field. The «rete mirabile» feeds
the gonads. Both thick muscular linings of the mesenterial vessels in
the trunk and the dorsal vessel in the opisthosome serve like a pump
additional to the heart. The blood envelope of the ventral blood vessel
in the trunk is seemed to provide the blood supply to the trophosome.
Through the use of 3D modeling of the circulatory system in the ten-
tacular crown of R. pachyptila we have discovered new vessels. Besides
the axial main afferent (and efferent) vessels known only for Riftia, there
are basal ones which are common for all vestimentiferans. The number
of the basal lamellar vessels is small; they are not required in the large R.
pachyptila. While the presence of the axial vessels is an achievement of
the large animal, the presence of basal vessels is an ancestral trait of
Vestimentifera. The presented features have phylogenetic importance,
since all these characters showed the direction of evolution of the organ
system in the vestimentiferan lineage of Siboglinidae.
The intravasal body varies its structure in the dorsal vessel,
depending on function. One is the hematopoietic organ, and another
one is valve-like structure preventing the backflow from the dorsal ves-
sels to the mesenterial vessel in the posterior most part of the trunk.
The structure of the sinus valvatus is significantly different in com-
parison to what was suggested earlier: the “valves” of the sinus formed
by paired coelomic cavities. Details of its ultrastructure we publish in
the following manuscript.
AUTHOR CONTRIBU TIONS
NNR-K and VM designed the study. SG collected and fixed the
material. NNR-K and VM performed the experiments, analyzed the
data, made illustrations and drafted the manuscript. All authors cor-
rected and approved the final version of the manuscript.
ACKNOWLEDGMENTS
We thank the crews of RV Akademik Mstislav Keldysh and manned
submersibles PISCES, MIR-1 and MIR-2 especially Dr. Anatoliy Saga-
levich for their brave underwater manipulations and collections of
the valuable material as vestimentiferan tubeworms. Galina Salkova
and Olga Matyukhina educated NNR-K in histological technique.
Elena Temereva and Leonid Rusin helped in mastering in 3D-
reconstruction software. Igor Popelyaev was a great inspirer for the
work. NR-K was supported in part by several grants. The histological
investigations were done with support by the Russian Science
RIMSKAYA-KORSAKOVA ET AL.
Foundation (#14-50-00029), the 3D modeling was supported by the
Russian Foundation for Basic Research (#14-04-32176 mol_a). The
analysis of the data was made by the Russian Science Foundation
(#14-14-00262). VM was supported in analysis of the results of the
work by the Russian Science Foundation (#14-50-00034). SG was
supported by the Russian Science Foundation (#14-50-00095).
L I T E RAT URE CI TE D
Andersen, A. C., Jolivet, S., Claudinot, S., & Lallier, F. H. (2002). Biometry
of the branchial plume in the hydrothermal vent tubeworm Riftia
pachyptila (Vestimentifera; Annelida). Canadian Journal of Zoology, 80,
320–332.
Andersen, A. C., Hourdez, S., Marie, B., Jollivet, D., Lallier, F. H., &
Sibuet, M. (2004). Escarpia southwardae sp. nov., a new species of
vestimentiferan tubeworm (Annelida, Siboglinidae) from West African
cold seeps. Canadian Journal of Zoology, 82, 980–999.
Arp, A. J., & Childress, J. J. (1983). Sulfide binding by the blood of the
hydrothermal vent tube worm Riftia pachyptila. Science, 219, 295–297.
Black, M. B., Halanych, K. M., Maas, P. Y., Hoeh, W. R., Hashimoto, J.,
Desbruyères, D., . . . Vrijenhoek, R. C. (1997). Molecular systematics
of vestimentiferan tubeworms from hydrothermal vents and cold-
water seeps. Marine Biology, 130, 141–149.
Bosch, C., & Grasse, P. P. (1984). Cycle partiel des bacteries chimioauto-
trophes symbiotiques et leurs rapports avec les bacteriocytes chez
Riftia pachyptila Jones (Pogonophore Vestimentifere). I. Le tropho-
some et les bacteriocytes. Compte Rendu Lebdomaire des seances
l’academie des Sci Paris (Ser III) 299, 371–376.
Braunbeck, T., & Dales, R. P. (1985). The ultrastructure of the heart-
body and extravasal tissue in the polychaete annelids Neoamphitrite
figulus and Arenicola marina. Journal of the Marine Biological Associa-
tion of the United Kingdom, 65, 653–662.
Bright, M., & Sorgo, A. (2003). Ultrastructural reinvestigation of the
trophosome in adults of Riftia pachhyptila (Annelida, Siboglinidae).
Invertebrate Biology, 122, 347–368.
Brusca, R. C., & Brusca, G. L. (2003). Invertebrates (p. 875). Sunderland
MA: Sinauer Associates.
Cardona, A., Saalfeld, S., Schindelin, J., Arganda-Carreras, I., Preibisch, S.,
Longair, M., . . . Douglas, R. J. (2012). TrakEM2 software for neural
circuit reconstruction. PLoS One, 7, e38011.
Cavanaugh, C. M., Gardiner, S. L., Jones, M. L., Jannasch, H. W., &
Waterbury, J. B. (1981). Prokaryotic cells in the hydrothermal vent
tube worm Riftia pachyptila Jones: Possible chemoautotrophic sym-
bionts. Science, 213, 340–342.
Dales, R. P., & Pell, J. S. (1970). Cytological aspects of haemoglobin and
chlorocruorin synthesis in polychaete Annelids. Zeitschrift fu€r Zellfor-
schung und mikroskopische Anatomie, 109, 20–32.
Eichinger, I., Hourdez, S., & Bright, M. (2013). Morphology, microanatomy
and sequence data of Sclerolinum contortum (Siboglindae, Annelida) of
the Gulf of Mexico. Organisms Diversity & Evolution, 13, 311–329.
Felbeck, H. (1981). The chemoautotrophic potential of the hydrothermal
vent tube worm, Riftia pachyptila Jones (Vestimentifera). Science, 213,
336–338.
Fransen, M. (1988). Coelomic and vascular systems. In W. Weistheide &
C. O. Hermans (Eds.), The ultrastructure of Polychaeta, Microfauna
Marina 4, 199–213.
Friedman, M. M., & Weiss, L. (1980). An electron microscopic study of
hemoglobin synthesis in the marine annelid, Amphitrite ornata (Poly-
chaeta: Terebellidae). Journal of Morphology, 164, 121–138.
| 17
Gardiner, S. L. (1992). Polychaeta: General organization, integument,
musculature, coelom, and vascular system. In F. W. Harrison & S. L.
Gardiner (Eds.), Annelida: Microscopic anatomy of invertebrates (Vol. 7,
pp. 19–52). New York: Wiley-Liss, Inc.
Gardiner, S. L., & Jones, M. L. (1993). Vestimentifera. In F.Harrison & M.
Rice (Eds.), Onychophora, chilopoda, and lesser protostomata: Micro-
scopic anatomy of invertebrates (Vol. 12, pp. 371–460). New York:
Wiley-Liss, Inc.
Glover, A. G., Källstro€ m, B., Smith, C. R., & Dahlgren, T. G. (2005).
World-wide whale worms? A new species of Osedax from the shal-
low north Atlantic. Proceedings of the Royal Society of London B Bio-
logical Sciences, 272, 2587–2592.
Glover, A. G., Wiklund, H., Taboada, S., Avila, C., Cristobo, J., Smith, C.
R., . . . Dahlgren, T. G. (2013). Bone-eating worms from the Antarctic:
The contrasting fate of whale and wood remains on the Southern
Ocean seafloor. Proceedings of the Royal Society of London B Biological
Sciences, 280, 1–10.
Halanych, K. M., Feldman, R. A., & Vrijenhoek, R. C. (2001). Molecular
evidence that Sclerolinum brattstromi is closely related to vestimenti-
ferans, not to frenulate pogonophorans (Siboglinidae, Annelida). Bio-
logical Bulletin, 201, 65–75.
Ivanov, A. V. (1960). Pogonophora (The Pogonophorans), Fauna of the
USSR, new series. Academy of. ed. Moscow, Leningrad.
Jones, M. L. (1980). Riftia pachyptila, New Genus, New Species, The Ves-
timentiferan Worm from the Galapagos Rift Geothermal Vents
(Pogonophora). Proceedings of the Biological Society of Washington, 93,
1295–1313.
Jones, M. L. (1981). Riftia pachyptila Jones: Observations on the Vesti-
mentiferan Worm from the Galapagos Rift. Science, 213, 333–336.
Jones, M. L. (1985). On the vestimentifera, new phylum: Six new species
and other taxa, from hydrothermal vents and elsewhere. Bulletin of
the Biological Society of Washington, 6, 117–158.
Jouin-Toulmond, C., Augustin, D., Desbruyeres, D., & Toulmond, A.
(1996). The gas transfer system in alvinellids (Annelida Polychaeta,
Terebellida). Anatomy and ultrastructure of the anterior circulatory
system and characterization of a coelomic, intracellular, haemoglobin.
Cahiers de Biologie Marine, 37, 135–151.
Karaseva, N. P., Malakhov, V. V., & Galkin, S. V. (2012). The morphology
and anatomy of the vestimentiferan worm Oasisia alvinae Jones,
1985 (Annelida: Siboglinidae). III. Coelomic cavity, trophosome and
blood, excretory and reproductive systems. Russian Journal of Marine
Biology, 38, 122–138.
Karaseva, N. P., Rimskaya-Korsakova, N. N., Galkin, S. V., & Malakhov, V.
V. (2016). Taxonomy, geographical and bathymetric distribution of
vestimentiferan tubeworms (Annelida, Siboglinidae). Zoologicheskii
Zhurnal, 95, 624–659.
Kennedy, G. Y., & Phillips Dales, R. (1958). The function of the heart-
body in polychaetes. Journal of the Marine Biological Association of the
United Kingdom, 37, 15–31.
Kojima, S., Ohta, S. T., Yamamoto, T., Yamaguchi, T., Miura, T., Fujiwara,
Y., . . . Hashimoto, J. (2003). Molecular taxonomy of vestimentiferans
of the western Pacific, and their phylogenetic relationship to species
of the eastern Pacific III. Alaysia-like vestimentiferans and relation-
ships among families. Marine Biology, 142, 625–635.
Li, Y., Kocot, K. M., Schander, C., Santos, S. R., Thornhill, D. J., & Hala-
nych, K. M. (2015). Mitogenomics reveals phylogeny and repeated
motifs in control regions of the deep-sea family Siboglinidae (Anne-
lida). Molecular Phylogenetics and Evolution 85, 221–229.
Li, Y., Kocot, K. M., Whelan, N. V., Santos, S. R., Waits, D. S., Thornhill,
D., & Halanych, K. M. (2016). Phylogenomics of tubeworms
 18 | RIMSKAYA-KORSAKOVA
(Siboglinidae, Annelida) and comparative performance of different
reconstruction methods. Zoologica Scripta, 1–14.
Malakhov, V. V., & Galkin, S. V. (1998). Vestimentifery— beskishechnye
bespozvonochnye morskikh glubin (Vestimentiferans—The Gutless Inver-
tebrates from Marine Depths). Moscow: KMK Scientific Press Ltd.
Malakhov, V. V., Popelyaev, I. S., & Galkin, S. V. (1996). Microscopic
anatomy of Ridgeia phaeophiale Jones, 1985 (Pogonophora, Vesti-
mentifera) and the problem of the position of Vestimentifera in the
system of the animal kingdom: III. Rudimentary digestive system,
trophosome, and blood vascular system. Russian Journal of Marine
Biology, 22, 189–198.
Mane-Garzon, F., & Montero, P. (1985). On a new type of tubicolous
worm: Lamellibrachia victori n.sp. (Vestimentifera). Pro- posal of a
new phylum: Mesoneurophora. Revista de Biologie (Uruguay), 8, 1–28.
Mangum, C. P., & Dales, R. P. (1965). Products of haem synthesis in pol-
ychaetes. Comparative Biochemistry and Physiology, 15, 237–257.
Meyer, E. (1887). Studien ueber den Korperbau der Anneliden. Mitth
Zool Stat Neapel, VII.

, G. (2004). Biochemical and enzymological aspects of
Minic, Z., & Herve
the symbiosis between the deep-sea tubeworm Riftia pachyptila and
its bacterial endosymbiont. European Journal of Biochemistry, 271,
3093–3102.
Pilger, J. F. (1993). Echiura. In F. W. Harrison & S. L. Gardiner (Eds.),
Onychophora, Chilopoda, and Lesser Protostomata: Microscopic anat-
omy of invertebrates (Vol. 12, pp. 185–236). New York: Wiley-Liss.
Potswald, H. E. (1981). Abdominal segment formation in Spirorbis moerchi
(Polychaeta). Zoomorphology, 97, 225–245.
Rimskaya-Korsakova, N. N., & Malakhov, V. V. (2010). Organization of
the Tentacular Apparatus of the Vestimentiferan Tubeworm Riftia
pachyptila, Jones 1981 (Annelida, Vestimentifera). Doklady Biological
Sciences, 433, 257–260.
Rimskaya-Korsakova, N. N., Malakhov, V. V., & Galkin, S. V. (2011).
Organization of the tentacular region of the vestimentiferan tube-
worm Riftia pachyptila 1981 (Annelida, Vestimentifera). Biological Bul-
letin, 38, 667–678.
View publication stats
ET AL.
Rouse, G. W. (2001). A cladistic analysis of Siboglinidae Caullery, 1914
(Polychaeta, Annelida): Formerly the phyla Pogonophora and Vesti-
mentifera. Zoological Journal of the Linnean Society, 132, 55–80.
Rouse, G. W., Wilson, N. G., Worsaae, K., & Vrijenhoek, R. C. (2015).
Report a dwarf male reversal in bone-eating worms. Current Biology,
1–6.
Schulze, A. (2002). Histological and ultrastructural characterization of the
intravasal body in Vestimentifera (Siboglinidae, Polychaeta, Annelida).
Cahiers de Biologie Marine, 43, 355–358.
Schulze, A., & Halanych, K. M. (2002). Siboglinid evolution shaped
by habitat preference and sulfide tolerance. Hydrobiologia, 496,
199–205.
Southward, E. C., Schulze, A., & Gardiner, S. L. (2005). Pogonophora
(Annelida): Form and function. Hydrobiologia, 535-536, 227–251.
Van der Land, J., & Nørrevang, A. (1975). The systematic position of
€r Zoological
Lamellibrachia (Annelida, Vestimentifera). Zeitschrift fu
System und Evolution, 1, 86–101.
Van der Land, J., & Nørrevang, A. (1977). Structure and Relationships
of Lamellibrachia (Annelida, Vestimentifera). Danske videnskabernes
Selskab, 21, 1–102.
Webb, M. (1969). Lamellibrachia barhami gen. nov. sp. nov., (Pogono-
phora) from the northeast Pacific. Bulletin of Marine Science, 19, 18–
47.
Zhadan, A., Vortsepneva, E., & Tzetlin, A. (2015). Ontogenetic develop-
ment and functioning of the anterior end of Cossura pygodactylata
Jones, 1956 (Annelida: Cossuridae). Zoomorphology, 134, 509–521.
How to cite this article: Rimskaya-Korsakova NN, Galkin SV,
Malakhov VV. The anatomy of the blood vascular system of the
giant vestimentiferan tubeworm Riftia pachyptila (Siboglinidae,
Annelida). Journal of Morphology. 2017;00:000–000. https://doi.
org/10.1002/jmor.20677