LAB

4 ANIMAL DIVERSITY III: PHYLA MOLLUSCA

AND ANNELIDA.

INTRODUCTION
This laboratory will be your first examination of
the coelomate phyla. An increase in structural
complexity over the acoels and pseudocoels will
be evident.
EXERCISE 4.1 Phylum Mollusca.
MATERIALS NEEDED:
Preserved Mytilus or other bivalve.
Preserved representatives for each class of
Mollusca and cleaned shells from each class.
Dissecting kit, compound microscope, glass
plates, finger bowls.
The Phylum Mollusca is second in size only to
the Phylum Arthropoda. Although members of
this phylum are extremely diverse in structure,
physiology, and habits, they are built on a
uniform body plan consisting of a mantle, shell,
and foot. The mantle secretes the shell and is
sometimes involved in respiration. The shell
provides support, protection, and secure
attachments for muscles. The foot is fleshy and
contains most of the viscera.
PROCEDURE:
1. Ciliary-mucoid Feeding in Clams. Hands,
claws, and teeth are not the only ways animals
collect and ingest food. Some rely on suckers
(flukes and leaches, for example), while others
have specialized mouthparts (the chewing,
sucking, siphoning and sponging mouthparts of
insects). In this portion of the laboratory you
will observe the processes of ciliary-mucoid
feeding by a clam. Clams are in the phylum
Mollusca, class Bivalvia, a trait they share in
common with other tasty beasts such as mussels,

scallops, and oysters. Bivalves have two lateral

shells (valves) hinged at their dorsal surface to
enclose the body (hence the name "bivalvia").
The gills (ctenidia) are unusually large and are
used for both respiration and as a food-gathering
device. Parts of the posterior mantle are
modified to form excurrent and incurrent
siphons (through which water is pumped to
ventilate and feed the animal). Most bivalves are
adapted to a burrowing lifestyle while others
may attach to a substratum by with tough byssal
threads. Some swim by rapidly clapping their
shells together (scallops, for example).
2. Obtain a clam from your instructor and orient the
animal so that the umbo (a swelling near the
hinge) is at the top-left of the pan (Fig 4.1). Your
clam is now lying on its right side with the
anterior toward the left, posterior at the right,
dorsal surface at the top and ventral surface at
the bottom of the pan (make sure it looks like
the diagram so you don't get confused). The two
largest muscles in the clam are the anterior and
posterior adductor muscles (Fig 4.1). The
adductor muscles work together to hold the
valves of the clam closed. The hinge is elastic,
so to open the valves the clam need only relax
the adductors. The anterior and posterior
retractor muscles are located near the adductors
and are used to draw the foot back into the shell.
To extend the foot out of the shell the clam fills
it with blood.
CAUTION: If you do the clam preparation,
be careful cutting the adductor muscles (Fig
4.1). Do not slice toward your palm or other
favorite body parts because the clam may
suddenly open or scalpel may slip.

ANTERIOR PEDAL
RETRACTOR

UMBO

CTENIDIA

POSTERIOR PEDAL
RETRACTOR

POSTERIOR
ADDUCTOR
ANUS

ANTERIOR
ADDUCTOR

EXCURRENT
SIPHON

MOUTH
PALPS

INCURRENT
SIPHON
FOOT
SHELL

MANTLE

Figure 4.1. Structures associated with feeding in the clam.

3. Insert the scalpel blade between the shells,
keeping it against the top valve (the clam's left
shell). Use a sawing motion to cut both the
anterior and posterior adductor muscles (locate
them by feel; Fig 4.1). Keep the blade against
the upper shell to avoid damaging internal
organs. When both adductors are cut the clam
will easily open. The thin flap of tissue lying
against the shell is the mantle. Note how the
mantle is thickened at the posterior end. These
structures lie against the similar thickenings on
the other shell mantle to form excurrent and
incurrent siphons (Fig 4.1). Water and
microorganisms are pumped through the
incurrent siphon by ciliary movements. The
water then flows over the gills (ctenidia) where
oxygen is exchanged for carbon dioxide. Mucusproducing cells coat the surface of the gills with
a sticky trap to capture microorganisms in the
flowing water. Cilia propel the sheet of mucus
toward the base of the gill where it is rolled into
fine threads or spheres (clam boogers?). The
mucus with trapped microorganisms is propelled
forward by cilia to oral palps that manipulate
and direct it into the mouth. After a short trip
through the esophagus, the material ends up in
the stomach. Most of the digestive system is
housed within the muscular foot of the clam
(we'll get back to that in a bit).

4. Remove the upper (clam's left) valve with a

twisting motion and expose the mother-of-pearl
(nacre) beneath the mantle. Pearls are formed
from this substance when sand grains fall
between the mantle and shell. The clam
responds to this irritation by depositing nacre
around the foreign object, thus building a pearl.
Search carefully though the mantle and you may
find a pearl. If the irritant was against the shell,
the pearl may be attached to the nacre or may
only be a lump on the inner surface of the shell.
Compare the nacre to the outer leathery
periostacum. Several calcarious layers are
sandwiched between the periostracum and inner
nacre (your instructor may have cross sections
of shells for viewing). Put the left valve aside
and turn your attention to the clam.
5. Use your scalpel to cut a flap in the foot to
expose the digestive organs (as in Fig 4.2). Don't
worry if your section does not show off the
internal structures as clearly as in the figure (this
is largely a matter of luck). Identify the stomach
and intestine by location and size. A spongy
digestive gland surrounds the stomach area
while a spongy gonad (ovary) occupies most of
the foot. Use forceps to gently tear the tissue
away from the dorsal surface of the clam to
expose the heart. If you were careful you can
identify both the auricle and ventricle.

A
T

I
J

L
P

A) UMBO

H) POSTERIOR ADDUCTOR

O) INTESTINE

B) DIGESTIVE GLAND

I) ANUS

P) FOOT

C) STOMACH

J) EXCURRENT SIPHON

Q) PALPS

D) PERICARDIUM

K) INCURRENT SIPHON

R) MOUTH

E) VENTRICLE

L) MANTLE

S) ANTERIOR ADDUCTOR

F) AURICLE

M) GILL

T) ESOPHAGUS

G) KIDNEY

N) DIGESTIVE GLAND

Figure 4.2. Internal structure of a clam. See text for details.

Note how the intestine passes through the center of the ventricle. Expose the kidney (located ventral to
the heart) and trace the remainder of the intestine to the anus (near the excurrent siphon. Nitrogenous
wastes from the kidney, carbon dioxide from the gills, and feces are expelled at the excurrent siphon.
7. Classes in the Phylum Mollusca. The phylum Mollusca is divided into seven classes (two, the
Aplacophora and Monoplacophora are not covered here). Examine the preserved and living specimens
as you work through this section. Compare your examples of each class to those of Figure 4.3. Record
your observations/sketches of class representatives in the results section.

Class Bivalvia. Members of this class include clams, mussels, scallops, and oysters (this class is also
known as Class Pelecypoda or Lamellibranchia). We've already covered this class in some detail, so
we'll concentrate on examples for the other classes.

B
A

Figure 4.3. Representative of the Phylum Mollusca. (A) A clam, Class Bivalvia; (B) A snail, Class
Gastropoda; (C) A tooth shell, Class Scaphopoda; (D) An octopus, Class Cephalopoda; and (E) a
chiton, Class Polyplacophora.

Class Gastropoda. This is the largest and

most widely-distributed of the molluscan
classes. The name "gastropoda" literally
means "stomach-foot" (these animals crawl
around on their stomach). Examples of
gastropods include snails, abalone, limpets,
nudibranchs, and slugs. While the snails,
abalone, and limpets have retained their
shells, those of nudibranchs and slugs have
been secondarily lost. Snails typically have
spiral shells, sometimes with an operculum
(a "trap door" used to close off the entrance
of the shell when the snail withdraws to the
interior). Abalone shells superficially
resemble a single clamshell. Their shell,
however, has a row of respiratory holes
along one edge. Limpet shells are conical
with a hole at the top (also an adaptation for
respiration). Nudibranchs are shell-less
marine forms with exposed gills (their name
means "naked gill"); some are extremely

colorful. Slugs, for our purposes, are snails

without shells. Gastropods are found on land
and in both fresh and salt-water habitats.
Depending on the species, they may be
predacious, herbivorous, or scavengers. A
few are adapted for suspension feeding.
Gastropods, like other mollusks, have a
peculiar file-like tongue called a radula. If
living freshwater snails are available, you
may be able to observe the use of the radula
as the animals feed on algae encrusted on
the sides of the tank.
Class Polyplacophora (= Amphinuria).
Polyplacophorans (chitons) have shells
composed of eight overlapping plates that at
least partially covered by the mantle. They
are almost prehistoric in appearance. Most
feed on algae and other encrusting
organisms by scraping them off the surface
of rocks with a radula. They are common
inhabitants of the intertidal zone. Examine

the preserved or living chitons on display.

What are the major structural differences
between these beasts and a shell-less
gastropod?
Class Scaphopoda. Scaphopods are a small
group of mollusks commonly known as "tooth
shells" or "tusk shells". They are rarely seen
alive since they're relatively small and burrow
in the sediment. Microorganisms make up the
bulk of their diet. Adhesive tentacles
sometimes capture larger food. Tooth shells are
all marine. Examine the scaphopod shells on
display.
Class Cephalopoda. Octopods, squid,
cuttlefish, and nautili are examples of
cephalopods (the term "Cephalopoda" literally
means "head-foot"). Most of the adaptations
seen in the cephalopods are related to their
swimming and predatory behavior. The mouth
is surrounded by a circlet of tentacles (or
arms). The shell is usually internalized (squid
pens and the cuttlebone of cuttlefish) or absent
(Octopus). Only the nautili and fossil forms
retain a shell. That of the nautili is chambered
and used both for protection and control of
buoyancy (compare a sectioned nautilus shell
to a gastropod shell, if available). The shells of
the now-extinct ammonoids were extremely
thick with complex sutures (some fossilized
specimens may be on display). Cephalopods

are voracious carnivores. Their eyes are among

the best of the invertebrates and eight or more
arms can quickly encircle prey. A parrot-like
beak with poison glands subdues and tears
prey. Clams, snails, crustaceans, and/or fish
may be eaten, depending on the species.
Cephalopods are restricted to marine waters.
EXERCISE 4.2 Phylum Annelida.
MATERIALS NEEDED:
Preserved animals: Class Oligochaeta: As
available; Class Polychaeta: clamworms
(Nereis sp); Class Hirudinea: leeches. Others,
as available.
Living animals: Aeolosoma, Dero, or Stylaria
sp., Earthworms (Lumbricus terrestris)
Glass plates, dissecting kits, pans.
Annelids (segmented worms) are
extremely successful and have invaded
terrestrial, freshwater, and marine habitats. They
include familiar forms such as earthworms and
leeches and many diverse marine organisms.
The segmentation characteristic of this phylum
extends to the interior of the animal and is more
complex than the annulations of the rotifera and
nematodes. Metamerism, a repetition of similar
body parts, is related to the segmentation and is
probably an adaptation for locomotion.

B
A
C

E
D

A) TENTACLES

D) PARAPODIA

B) SENSORY PAPILLAE

E) JAWS

C) EYES

F) EVERTED PHARYNX

Figure 4.4. Anterior segments of the clamworm, Nereis (Phylum Annelida, Class Polychaeta).

PROCEDURE:
1. A Survey of Segmented Worms. Examine the
living and preserved material on display and
identify to their class. Record your observations
and/or sketches of representative animals.
Class Oligochaeta. Members of the Class
Oliogochaeta include earthworms and many
freshwater forms. Metamerism is welldeveloped and most apparent in the posterior
segments. The head is degenerate with few
obvious sensory structures (owing to their
burrowing habits). Oligochaetes have few
setae (spine-like appendages; oligo="few",
chaeta="setae"). To observe setae, prepare a
wet mount of an aquatic oligochaete
(Aeolosoma, Dero, and/or Stylaria sp.) and
view under scanning or low power. The setae
are the spine-like hairs extending from the
body (diagram and label a typical individual).
After observing your specimen for a short
period, what do you think might be the
function of the setae? Your friendly
neighborhood earthworms also have setae, but
theirs are considerably smaller than those of
aquatic forms. Members of this class also
possess a reproductive structure called the
clitellum (covered later). Oligochaetes are
monoecious.
Class Polychaeta. Polychaetes are the most
structurally diverse of the annelids and are
exclusively marine. They may be sedentary or
free moving. The free-moving forms have the
more typical polychaete body plan while the
sedentary and tube-dwelling forms are often
bizarre in the extent of their modifications.
Polychaetes have many setae (poly="many")
and gill- or fin-like appendages called
parapodia (Fig 4.4). In some species the setae
may cover the entire body which gives it a
furry appearance (sea mouse, Aphrodite).
Others have hollow poison-tipped setae used
for defense (as in the "fire worms"). The
parapodia may be relatively simple (Nereis),
or extremely complex and varying from
segment to segment (Chaetopterus). The
heads of polychaetes are usually more
complex than those of oligochaetes. Eyes,
tentacles, palps, and pincer-like jaws are the
rule. The heads of plume and feather duster
worms are endowed with tentacles to capture
prey. Polychaetes lack a clitellum and are
mostly dioecious. Remove a clamworm

(Nereis) from its container and examine under

a dissecting microscope. Identify the tentacles,
sensory papillae, eyes, and parapodia (Fig
4.4). Pulling gently on the jaws with forceps
can evert the pharynx (otherwise, the head
must be cut so it can be exposed).
Class Hirudinea. This freshwater class of
annelids is commonly called leeches. They
usually have an anterior sucker surrounding
the mouth and a ventrally-facing posterior
sucker. Like earthworms they are monoecious,
but asynchronously so (individuals switch
between being male or female, but are never
both at the same time). Although leeches have
a reputation as being blood suckers, most are
predacious.
2. Observations of Living Earthworms
(Lumbricus terrestris). Obtain a living
earthworm from its culture and place in a finger
bowl. Locate the conspicuous clitellum, a
saddle-like swelling on the dorsal surface. The
clitellum has a high concentration of mucussecreting glands. It produces a mucus sheath
used to surround worms during mating and is
responsible for making the cocoon within which
fertilized eggs are deposited. The ventral side of
the earthworm is usually lighter in pigmentation
than the dorsal. If you run your finger back and
forth along the ventral surface, you will feel the
setae (which way do they point? If the animal
won't hold still for this, have your lab partner
trap the worm under a paper towel). Place the
earthworm first on a glass plate, then some
paper toweling, and gently prod it into motion.
What earthworm appendages are affected by the
nature of the substrate?
3. Gross Anatomy of Lumbricus terrestris, an
Oligochaete. Anesthetize an earthworm by
immersing it in 70% ethanol. Note the
thickening of the mucus coat as the animal
moves around. Does this suggest a function for
the mucus coat? What other functions would you
attribute to the coat? As soon as the animal
quiets down, remove it from the ethanol to avoid
over-anesthetization and transfer to a dissecting
pan. The anterior of the animal is the end closest
to the clitellum (it is more cylindrical than the
flattened posterior). Locate the prostomium, the
first lobe at the anterior end. The mouth is
located directly behind the prostomium on the
ventral surface. For convenience in locating
body structures, the segments of the earthworm

10

A
B
C

A) MOUTH
B) BRAIN

C) CIRCUM-PHARNGEAL
CONNECTIVE

D) PHARYNYX

E) ESOPHAGUS
F) "HEARTS" (5 PAIR)

G
15

H
I

J) GIZZARD

K) INTESTINE

30

35

40
45

H) SEMINAL VESICLES
I) CROP

20

25

G) SEMINAL RECEPTACLES

L) DORSAL BLOOD
VESSEL
M) SEPTA
N) NEPHRIDIA

O) ANUS

N
O

FIGURE 4.5. MAJOR INTERNAL ORGANS

OF AN OLIGOCHAETE (Lumbricus terrestris).
See the text for a discussion of their functions.
are numbered starting at the segment directly
behind the mouth (the prostomium is segment
zero by convention). Refer to these numbers
during your dissection (Figure 4.5). Turn your
earthworm over (dorsal side up) and position it
along one edge of the dissecting pan so you can
view your specimen under a dissecting

microscope. Pin it down through the

prostomium, then further back (anywhere behind
the clitellum would be fine). Cut a slit in the
dorsal surface near the posterior pin. Using
scissors or a scalpel (whichever you are most
comfortable with), extend the cut forward to the
first segment. Be careful not to cut too deep.
Starting at segment one, use a scalpel to cut the
septa that internally divide the segments (Figure
4.5). Cut the septa so that the skin can be laid
flat. Use additional pins to hold the integument
open and expose the organs. Whenever possible,
place the pins in every fifth or tenth segment;
this allows you to easily locate organs without
having to recount. Continue to lay the skin back
until you have uncovered a centimeter or so of
the intestine. Starting at the anterior end, locate
the muscular pharynx (note the threadlike
muscles connected to the body wall). This is
followed by a totally tubular esophagus, which
terminates in a crop (the wider organ near
segment 15). Posterior to the crop you will find
the gizzard. Gently press on the crop and
gizzard to test their firmness. While the crop is
soft and thin, the gizzard is very muscular (soil
is ground up and churned within the gizzard).
The gizzard is followed by a long intestine.
Reproductive structures start at segment nine.
Identify the seminal receptacles (bulblike
organs in segments nine and ten) and seminal
vesicles
(segments
9,10,11,12).
Other
reproductive organs are usually difficult to
identify in preserved specimens. These include
the testis (segments 10 and 11), seminal funnels
(opening at 10 and 11 and extending as sperm
ducts to 15) and ovaries (segment 15). Attempt
to locate these structures in your specimen. Use
your text to determine the function of each
(record in the Results Section). The circulatory
system is visible as a dorsal blood vessel
running along the upper surface of the digestive
system (a ventral vessel is hidden on the lower
side; Figure 4.6). The dorsal and ventral vessels
are
connected
by
five
pairs
of
circumesophageal vessels (hearts) starting at
segment seven. Examine your animal under a
hand lens or dissecting microscope to locate the
nephridia in each body segment (they are easier
to find near the intestine). The nephridia act as
kidneys to remove wastes that collect in the
coelomic fluid (in the body cavities between the
septa). Locate the white (gray?) lobes of the

brain (or cerebral ganglion) at the anterior end

(in front of the pharynx). A pair of nerves (the
circumpharyngeal connectives) surrounds the
pharynx and leads to the ventral nerve cord.
Gently push the intestine aside to expose the
underlying ventral nerve cord. Within each
segment you will find a small knot of nervous
tissue (the segmental ganglion). Each ganglion
is responsible for controlling muscular
movements within its own segment. The ventral

Typhlosol

nerve cord allows ganglia to communicate

among themselves to coordinate their activity.
4. Microscopic Examination of an Earthworm.
Examine a cross section of an earthworm under
scanning and low power and locate the
structures indicated in Figure 4.6. Of what
functional significance is the arrangement of the
circular and longitudinal muscles in your
specimen? Why does the intestine have a
shelflike typhlosole running along its length?

Dorsal Blood Vessel

Cuticle

Gastrodermis

Epidermis
Circular Muscle

Chlorogogue
Cells

Longitudinal
Muscle

Setae

Coelom

Nephridium
Nephrostome

Subneural
Blood Vessel

Ventral
Nerve Cord

Ventral Blood Vessel

FIGURE 4.6. MAJOR INTERNAL ORGANS OF AN OLIGOCHAETE (Lumbricus terrestris). See

the text for a discussion of their functions.

REPORT SECTION___________________________________________
Lab 4 (Name) (Date/ Lab Section)
RESULTSANDDISCUSSION

EXERCISE 4.1 Phylum Mollusca. Classes in the Phylum Mollusca. Record your
sketchesand/orcommentsforeachofthemolluscanclassesinthefollowing
table.
Class
CommonName
Sketchesand/orComments
Bivalvia

Gastropoda

Scaphopoda

Cephalopoda

Polyplacopho
ra

EXERCISE 4.2 Phylum Annelida. A Survey of Segmented Worms. Record your

sketches and/or comments for each of the annelid classes in the following
table.
Class
CommonName
Sketchesand/orComments
OLIGOCHAETA

POLYCHAETA

HIRUDINEA

Makeasketchofafreshwateroligochaete(Aeolosoma, Dero,and/or Stylaria

sp.).
SPECIES:_________________
Fromyourobservations,whatdoyou
MAGNIFICATION:___________
thinkisthefunctionofthesetae?

_________________________________________
_________________________________________
_________________________________________
_________________________________________
_________________________________________
_________________________________________
_________________________________________

Discussthediversityinbodystructureseenamongtheclassesofannelids.
Howaretheseadaptationsrelatedtotheanimals'behavior,ecology,feeding,
andgenerallifestyle?
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GrossAnatomyofLumbricusterrestris,anOligochaete.
Consult your text (or another source) to briefly describe mating in the
earthworm
Lumbricus

terrestris.
______________________________________________________________________
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Use your text to complete the following table. In the Male/Female column,
indicateifthestructureispartofthemaleorfemalereproductivesystem.
Organ
Male/Femal
Function
e
Clitellum
_______________________________________________
_______________________________________________
_______________________________________________
_______________________________________________
______________________
Seminal
_______________________________________________
Receptacles
_______________________________________________
_______________________________________________
_______________________________________________
______________________
SeminalVesicles
_______________________________________________
_______________________________________________
_______________________________________________
_______________________________________________
______________________
Testis
_______________________________________________
_______________________________________________
_______________________________________________
_______________________________________________
______________________
SeminalFunnels
_______________________________________________
_______________________________________________
_______________________________________________
_______________________________________________
______________________
SpermDucts
_______________________________________________
_______________________________________________
_______________________________________________

Ovaries

_______________________________________________
______________________
_______________________________________________
_______________________________________________
_______________________________________________
_______________________________________________
______________________

MicroscopicExaminationofanEarthworm.Ofwhatfunctionalsignificanceis
thearrangementofthecircularandlongitudinalmusclesinyourspecimen?
_____________________________________________________________________________
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_____________________________________________________________________________
Whydoestheintestinehaveashelfliketyphlosolerunningalongitslength?
_____________________________________________________________________________
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_____________________________________________________________________________
Discussthediversityinbodystructureseenamongthethreeclassesof
annelids.Howaretheseadaptationsrelatedtotheanimals'behavior,ecology,
feeding,andgenerallifestyle?
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_____________________________________________________________________________
_____________________________________________________________________________
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