Culture of Marine Polychaetes
J. Sesh Serebiah
Introduction
The Polychaeta or polychaetes are a class of
annelid worms, generally marine. Each body seg-
ment has a pair of fleshy protrusions called
parapodia that bear many bristles, called chaetae,
which are made of chitin. Indeed, polychaetes are
sometimes referred to as bristle worms. More
than 10,000 species are described in this class.
Common representatives include the lugworm
(Arenicola marina) and the sandworm or clam
worm Nereis. Polychaetes as a class are robust
and widespread, with species that live in the
coldest ocean temperatures of the abyssal plain,
to forms which tolerate the extreme high
temperatures near hydrothermal vents.
They are segmented worms, generally less
than 10 cm (3.9 in.) in length, although ranging
at the extremes from 1 mm (0.039 in.) to 3 m
(9.8 ft). They are often brightly colored and may
be iridescent or even luminescent. Each segment
bears a pair of paddle-like and highly
vascularized parapodia, which are used for
movement and, in many species, act as the
worm’s primary respiratory surfaces. Bundles
of bristles, called setae, project from the
parapodia. However, polychaetes vary widely
from this generalized pattern and can display a
range of different body forms. The most
J.S. Serebiah (*)
Department of Marine Studies and Coastal Resources
Management, Madras Christian College, Tambaram,
Chennai, 600054, Tamil Nadu, India
e-mail: seshserebiah@yahoo.com
S. Perumal et al. (eds.), Advances in Marine and Brackishwater Aquaculture,
DOI 10.1007/978-81-322-2271-2_5, # Springer India 2015
generalized polychaetes are those that crawl
along the bottom, but others have adapted to
many different ecological niches, including
burrowing, swimming, pelagic life, tube-
dwelling or boring, commensalism, and parasit-
ism, requiring various modifications to their
body structure. The head, or prostomium, is rela-
tively well developed, compared with other
annelids. It projects forward over the mouth,
which therefore lies on the animal’s underside.
The head normally includes two to four pairs of
eyes, although there are some blind species.
These are typically fairly simple structures, capa-
ble of distinguishing only light and dark,
although some species have large eyes with
lenses that may be capable of more sophisticated
vision. The head also includes a pair of antennae,
tentacle-like palps, and a pair of pits lined with
cilia, known as “nuchal organs.” These latter
appear to be chemoreceptors and help the worm
to seek out food. The outer surface of the body
wall consists of a simple columnar epithelium
covered by a thin cuticle. Underneath this, in
order, there are a thin layer of connective tissue,
a layer of circular muscle, a layer of longitudinal
muscle, and a peritoneum surrounding the body
cavity. Additional oblique muscles move the
parapodia. In most species, the body cavity is
divided into separate compartments by sheets of
peritoneum between each segment, but in some
species, it is more continuous. The mouth of
polychaetes varies in form depending on their
diet, since the group includes predators,
herbivores, filter feeders, scavengers, and
43
44
Fig. 1 Parts and cross-sectional diagram of polychaete
parasites. In general, however, it possesses a pair
of jaws and a pharynx that can be rapidly everted,
allowing the worm to grab food and pull it into
the mouth. In some species, the pharynx is
modified into a lengthy proboscis. The digestive
tract is a simple tube, usually with a stomach
partway along. The smallest species and those
adapted to burrowing lack gills, breathing only
through their body surface. Most other species,
however, have external gills, generally, although
not always, associated with the parapodia. There
is usually a well-developed, if simple, circulatory
system. There are two main blood vessels, with
smaller vessels to supply the parapodia and the
gut. Blood flows forward in the dorsal vessel,
above the gut, and returns back down the body
in the ventral vessel, beneath the gut. The blood
vessels themselves are contractile, helping to
push the blood along, so most species have no
need of a heart. In a few cases, however, muscu-
lar pumps analogous to a heart are found in
various parts of the system. Conversely, some
species have little or no circulatory system at
all, transporting oxygen in the coelomic fluid
that fills their body cavity. The blood itself may
be colorless or have any of three different respi-
ratory pigments. The most common of these is
J.S. Serebiah
hemoglobin, but some groups have hemerythrin
or the green-colored chlorocruorin instead
(Fig. 1).
The nervous system consists of a single or
double ventral nerve cord running the length of
the body, with ganglia and a series of small nerves
in each segment. The brain is relatively large,
compared with that of other annelids, and lies in
the upper part of the head. An endocrine gland is
attached to the ventral posterior surface of the
brain and appears to be involved in reproductive
activity. In addition to the sensory organs on the
head, there may also be photosensitive eyespots
on the body, statocysts, and numerous additional
sensory nerve endings, most likely involved with
the sense of touch. Polychaetes have a varying
number of protonephridia or metanephridia for
excreting waste, which in some cases can be
relatively complex in structure. The body also
contains greenish “chloragogen” tissue, similar
to that found in oligochaetes, which appears to
function in metabolism, in a similar fashion to
that of the vertebrate liver. Their cuticle is
constructed from cross-linked fibers of collagen
and may be 200-nm to 13-mm thick. Their jaws
are formed from sclerotized collagen and their
setae from sclerotized chitin.
Culture of Marine Polychaetes
Fig. 2 Polychaete scoop
Sampling and Identification
Polychaetes in sandy/muddy sediments are sam-
pled using corers. They are collected by “poly-
chaete scoop” (Fig. 2). This is made from a
modified galvanized sheet metal dryer vent
(available at any good building supply store)
fastened to a garden hoe, as a handle. The
scoop is dragged along the sediment catching
the top polychaete-containing layer. The vent
has a hole covered with galvanized wire mesh,
which in turn is covered with fiberglass window
screening adhered with silicone caulk. This hole
allows water to pass through the scoop while the
mesh catches any polychaetes that swim out of
the sediment. The full scoop of sediment is then
sieved through two screens, an upper one with a
500-μm mesh and a lower one constructed of
fiberglass window screening (approximately
64-μm mesh size). The upper sieve catches larger
stones and macrobenthic organisms, which
retained the polychaetes. This operation is
repeated numerous times, resulting in a bucket
of concentrated polychaetes containing the
desired species. The sieving operation must be
performed gently, with the lower sieve
submerged in seawater so that the interstitial
organisms are not ground up in the sand or
against the mesh. Additional sediment is sieved
only through the coarse mesh, to remove rocks
45
and larger meiofauna. Once a sufficient quantity
of sediment has been sieved, the concentrated
polychaetes must be taken back to the laboratory
to separate the desired species from the other
interstitial species present. Identification of
polychaetes is performed by Day (1967) and
Kristian Fauchald (1977). This is important to
avoid the isolation of larvae of different species,
which might be difficult to distinguish from those
of the study species. In addition, it is necessary to
remove polychaetes that might feed on the study
species or that might outcompete the study spe-
cies for food. Transfer a small amount of sedi-
ment to a shallow petri dish for examination
under a low-power stereo microscope. The
worms hide in their sand-mucus tubes, but they
can be captured by agitating the tube with a wide-
bore Pasteur pipette until the worm swims out
and then sucking it into the pipette for transfer to
another dish. The worms are checked to make
sure they are the correct species before being
added to the culture vessel. The brief identifica-
tion chart and keys up to family are given here-
with for classification and identification of
polychaetes (Fig. 3 and Table 1).
Significance of Polychaetes Culture
The importance of polychaetes in aquaculture
industry is being understood now and caused
 Group
Animalia

Phylum
Annelida

Class Class class class

Polychaeta Clitellata Myzostomida Archiannelida

sub class subclass

Palpata Scolecida
(Family 13)

order order
Aciculata Canalipalpata

Eunicida Sabellida
(Family 10) (Family 6)

Phyllodocida Spionida
(Family 28) (Family 8)

Terebellida
(Family 13)

Fig. 3 Flowchart of polychaetes’ classification up to order

Table 1 Showing polychaetes identification key up to subclass and family

Head without pairedfeeding tentacles, Scolecida: A group of subsurface sediment eaters.

without jaws. Each segment with or Mostly found on soft shores. Few of this group lives
without a pair of gills on rocky shores.
Head with one pair of flexiable grooved Spionida: Tube or burrow dwelling surface particle
feeding tentacles without jaws. Anterior pickers. Mostly found on soft shores or subtidally,
segment often with a pair of gills Few of this group lives on rocky shores.
Head with many pairs of feeding Terebellida: A group of tube-dwelling particle
flexiable tentacles, without jaws. pickers. Mostly found on soft shores or in sediment
subtidally, and some of this group is common in
rock crevices.
Head usually with a terminal funnel like Sabellida: A group of tube dwelling particle
fan of inflexiable ‘tentacles’, without filterers. Some of this group lives in colonial group
jaws on rocky shores, some on soft shores. Most live
subtidally.
Head with Jaws up to two pincer- Phyllodocida: A group of mostly surface wandering
chitinous like pairs terminal on food graspers occurs everywhere. Some of this
jaws, usually extensible proboscis. group is well adapted to rock crevices.
(not Each segment without a
glyceridae, pair of gills.
Goniadidae) Jaws only one pincer Eunicida: A group of mostly burrowing food
also like pair, barely graspers , mostly subtidal, in sediment or on rock
conspicuous extensible, but grouped coral. A few of this group are well adapted to rock
eyes, and with other toothed crevices. Few soft species.
short sensory plates. Mid body
tentacles. segments often with pair
of gills.
Culture of Marine Polychaetes
rapid demand as live feed for varieties of
cultivable species. Hence the technique on
polychaete culture need to be standardized for
potential polychaete species. Polychaetes as a
live feed can stimulate gonad maturation during
spawning in hatchery-reared species, e.g., Solea
vulgaris, Solea senegalensis (Dinis 1986),
Penaeus kerathurus (Luis 1989), and Penaeus
vannamei (Lytle et al. 1990). The other main
point of the culture is to reduce the substrate-
harvesting disturbance and the great biogeo-
chemical and benthic community impact
(Gambi et al. 1994) as they are used as
commercial bait.
Culture of Polychaetes
Collections of Brood Stocks, Breeding,
and Development
Adult worms are collected from the habitat.
Adults are gravid from April or May to October.
The worms can be kept in room temperature
aerated aquaria in 32-ppt artificial seawater and
adults sexed according to the color of their game-
togenic parapodia. Adult male and female worms
are best kept in separate tanks to prevent prema-
ture spawning if possible. Separation of adult
male and female are easier in tube dwelling
polychaetes. The adults were kept inside their
tubes at all times, and access to the worm was
achieved by making a small longitudinal cut in
the tube. Longitudinal cuts are repaired quickly
by the worm. Transverse or chevron-shaped cuts
cause irreparable damage to the tube, decreasing
the ability of the adult to feed and survive.
Oocytes are obtained by cutting off a single
parapodium from a gravid female and emptying
the contents (several thousand eggs) into a
150-μm screen immersed in 0.2-μm filtered arti-
ficial seawater (FASW) at 32-ppt salinity in a
35-mm petri dish. Oocytes are washed through
this screen; retained mucous and parapodial
fragments are discarded. The sieved oocytes are
washed three more times by transferring them
into 100-mm petri dishes of fresh FASW and
then incubated in FASW for about 40 min before
47
fertilization to allow for germinal vesicle break-
down. Sperm is prepared by cutting off the para-
podium of a ripe male without allowing the
contents to mix with seawater. The “dry sperm”
is extracted with a Pasteur pipette and diluted
1:1,000 with FASW. This sperm suspension is
examined through a stereo microscope, and when
the sperm are seen to be motile, 1–3 drops are
added to the eggs in a 100-mm petri dish. Know-
ing the optimal amount of sperm suspension to
use to obtain complete fertilization without
excessive polyspermy is a matter of experience,
because the concentration of dry sperm varies
between animals and over time in the same ani-
mal. Swimming ciliated blastulae develop by
12 h at room temperature (21–23  C). These
blastulae may then be transferred to the culture
apparatus. In many interstitial polychaetes, fer-
tilization is internal and its embryos are brooded
inside the mother’s tube. This makes obtaining
early embryonic stages difficult, because
gametes are not easily harvested for in vitro fer-
tilization. Most of the tube-dwelling polychaetes
have early swimming larvae in the development
stages. They are normally swimming in the
water. Early-swimming larvae may then be
extracted from the culture water.
Simulated Beach
Simulated beach is developed for harboring adult
worms for culture. The tube-dwelling worms
construct vertical tubes in sand or clay, which
are barely visible to the naked eye, and
epibenthos can be seen on the surface. Population
peaks occur in late post-monsoon and early sum-
mer (Zajac 1991), making this the best time to
collect the adults. The “simulated beach” is made
by depositing about 3 cm of sediment from the
collection site into a wide shallow vessel, deep
enough to allow for 6–8 cm of water on top
(Fig. 4). The sediment should first be frozen to
kill any fauna remaining and then rinsed several
times with freshwater to remove some of the
dead organic material, which could produce
unwanted products of decomposition. After
deposition of the sediment, natural or artificial
48
Fig. 4 Simulated beach
Fig. 5 Ideal method for the changing of water from simulated beach set up
seawater (ASW) is added. The seawater should
be changed at least twice to dilute out any
remaining freshwater and to remove some of
the silty organic debris, which will wash out the
newly placed sediment. Washed autoclaved sand
may be layered on top if the sediment is too silty.
Once the substrate has settled, the sorted worms
can be scattered over the surface. They will col-
onize the sediment soon. Two operations are
necessary to maintain the culture long enough
to produce larvae. First, adult animals must be
fed. For the 30-cm culture vessel, we use 4 ml of
concentrated liquid invertebrate food diluted in
J.S. Serebiah
some ASW and broadcast evenly over the sedi-
ment once per week. Overfeeding should be
avoided to prevent excessive bacterial growth
due to decomposing food matter. Second, the
culture water must be changed regularly to pre-
vent waste products, bacteria, and unwanted vol-
unteer organisms from building up and to
retrieve any larval offspring that may have been
released into the seawater. Changing water by
siphoning the old water into a 63-μm sieve
(Fig. 5) so that any larvae will be caught can
drop once again to culture system. If any other
meiofauna are captured in the sieve, they may be
Culture of Marine Polychaetes
removed before the desired polychaete larvae are
transferred to the culture apparatus by gently
backwashing the sieve with ASW from a squeeze
bottle. Two changes of water per week should be
sufficient to maintain healthy cultures. The inlet
of the siphon tube may be passed back and forth
through the culture vessel to ensure that as many
larvae as possible are caught. Fresh seawater is
added through a side inlet so that the disturbance
of the sediments is minimized. If natural seawa-
ter is used, it may be necessary to filter out
suspended fauna, such as small crustaceans, that
might invade the colony. In our experience, lar-
vae begin to appear in the culture systems around
the 40th day after beginning the culture. How-
ever, the number of larvae retrieved varies over
time, with a periodicity of around 30 days, prob-
ably due to the breeding and life history
characteristics of the worms. In our hands, cul-
ture life has been 3 months or roughly 2 months
of larval production. After this time the adult
population declines and larval collections drop
off.
Food for Culture Organisms
Initially, each individual can feed by once in
every 3 days with commonly available commer-
cial food used for tropical fish. This food type
had been used successfully with polychaete spe-
cies (Garwood and Olive 1981; Bartels-Hardege
and Zeeck 1990). After 20–25 days, they have to
feed daily, because of cannibalism activity.
Attempts to use commercially available aquar-
ium food, such as the liquid invertebrate food or
dried rotifers, resulted in excessive bacterial
growth in the colonies and poor survival of the
polychaete. Several types of algae can be used,
but Dunaliella salina, a green alga, and
Isochrysis galbana, a golden-brown alga, give
adequate nutrition, and they are easy to maintain
in culture. Autoclaved half-liter widemouthed
glass bottles with cotton stoppers are used as
culture vessels to avoid bacterial or fungal
49
contamination for algal culture. The
supplemented 0.2-μm filtered artificial seawater
(FASW) with F/2 medium at standard concentra-
tion (Strathmann 1987) can act as good culture
medium.
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