Invertebrate Anatomy OnLine

Lumbricus terrestris ©
Earthworm
28may2007
Copyright 2003 by
Richard Fox
Lander University
Preface
This is one of many exercises available from Invertebrate Anatomy OnLine , an Internet
laboratory manual for courses in Invertebrate Zoology. Additional exercises, a glossary, and
chapters on supplies and laboratory techniques are also available at this site. Terminology and
phylogeny used in these exercises correspond to usage in the Invertebrate Zoology textbook by
Ruppert, Fox, and Barnes (2004). Hyphenated figure callouts refer to figures in the
textbook. Callouts that are not hyphenated refer to figures embedded in the exercise. The
glossary includes terms from this textbook as well as the laboratory exercises.

Systematics
Annelida P, Clitellata, Oligochaeta C, Haplotaxida O, Lumbricina sO, Lumbricoidea SF,
Lumbricidae F (Fig 13-7A)

Annelida P
Annelida consists of the segmented worms in the major taxa Polychaeta (bristleworms),
Oligochaeta (earthworms and relatives), Branchiobdellida (crayfish ectosymbionts), and Hirudinea
(leeches) with a total of about 12,000 known species in marine, freshwater, and terrestrial
environments. The segmented body is composed of an anterior prostomium, a linear series of
similar segments, and a posterior pygidium. The prostomium and pygidium are derived from
anterior and posterior ends of the larva whereas the intervening segments arise through mitotic
activity of mesodermal cells in the pygidium.
The body wall consists of a collagenous cuticle secreted by the monolayered epidermis. A
connective tissue dermis lies beneath the epidermis. The coelom is lined by a peritoneum which
may be specialized to form the body wall muscles. Most annelids have chitinous bristles, or
chaetae, secreted by epidermal cells, that project from the body. The coelom is large, segmentally
compartmented, lined by peritoneum, and well developed in polychaetes and oligochaetes but
reduced in leeches. Successive coelomic spaces are separated by transverse bulkheads known
as septa which consist of double layers of peritoneum with connective tissue in between. The right
and left sides of each segmental coelom are separated by longitudinal mesenteries which, like
septa, are double layers of peritoneum with connective tissue between.
The gut is a straight, regionally specialized tube that begins at the mouth at the anterior end
and extends for the length of the body to end at the anus on the pygidium. It penetrates each
septum and is supported by dorsal and ventral mesenteries. Like that of most invertebrates, the gut
consists of ectodermal foregut, endodermal midgut, and ectodermal hindgut. The nervous system
consists of a dorsal brain in or near the prostomium, a pair of circumpharyngeal connectives
around the anterior gut, and a double, ventral nerve cord with paired segmental ganglia and
nerves. The hemal system of most annelids is a set of tubular vessels, some of which are
contractile and serve as hearts. The hemal system is absent or greatly reduced in leeches. The
system includes a dorsal longitudinal vessel above the gut in which blood moves anteriorly, a
ventral longitudinal vessel below the gut, in which blood moves posteriorly, and paired segmental
vessels that connect the dorsal and ventral vessels. The digestive, hemal, and nervous systems
are continuous and pass through the segments.

Respiration is accomplished in a variety of ways. In some, the general body surface is

sufficient but gills are present in most polychaetes, many leeches, and a few
oligochaetes. Excretory organs are metanephridia or protonephridia and typically one pair is
present in each segment. These osmoregulatory organs are best developed in freshwater and
terrestrial species. The sexes are separate in polychaetes but oligochaetes and leeches are
hermaphroditic. In the ancestral condition paired submesothelial clusters of germ cells were
present in each segment and released developing gametes into the coelom. In derived taxa
reproductive functions tend to be confined to a few specialized genital segments. Gametes mature
in the coelom or its derivatives and fertilization is external. Gametes are shed through ducts
derived from metanephridia or by rupture of the body wall. Spiral cleavage follows fertilization.
Clonal reproduction is common.
Clitellata
Clitellata, the sister taxon of Polychaeta, includes earthworms and their allies and the
leeches. The head is reduced and its sensory functions minimized. Clitellates lack the parapodia
characteristic of polychaetes although chaetae may be present. Also lacking are head and
pygidial appendages. Present however, is a girdle-like band of secretory epidermis, the clitellum,
near the anterior end of the worm. The brain has moved out of the prostomium into a more
posterior segment. These worms are hermaphroditic and have gonads and reproductive equipment
restricted to a few specialized genital segments. Development is direct, without a larva.
Oligochaeta C
Most of the 3500 known species of oligochaetes are small annelids found in benthic aquatic
habitats, mostly freshwater, but about 200 species occur in the sea. The taxon also includes the
large terrestrial earthworms, some of which reach lengths of three meters.
Laboratory Specimens
Of the many terrestrial earthworms in Lumbricidae, Lumbricus terrestris, the night crawler, is
the best known, mostly because it is a favorite fishing bait. Because of its availability and large
size, it is a good subject for a laboratory study of oligochaete anatomy. Living worms are readily
available from bait shops and are easily anesthetized in 5-7% non-denatured ethanol in
tapwater. Worms should be placed in the anesthetic about 15-30 minutes prior to the beginning of
the laboratory period. Preserved worms may be used if necessary but the following description
applies specifically to fresh specimens. This is a rare instance in which living material is less
expensive and easier to obtain than preserved and it should be taken advantage of. Living,
anesthetized worms are far, far superior to preserved worms for the study of oligochaete anatomy.

External Anatomy
Study an anesthetized living Lumbricus in a dissecting pan of 5-7% ethanol on the stage of
the dissecting microscope. The liquid must be deep enough to completely cover the worm.
Orientation
Examine the external features of the worm (Figs 1, 2). The anterior end is usually larger
than the posterior and is round in cross section whereas the posterior tends to be flattened
dorsoventrally. The dorsum is dark and rounded while the venter is pale and slightly
flattened. The dark dorsal blood vessel can be seen through the body wall on the dorsal
midline. Find right-left, dorsal-ventral, anterior-posterior, and the plane of symmetry.
 Figure 1. A dorsal view of the anterior end of Lumbricus. The segments are
numbered. Oligo54L.gif

Clitellum
A band of thickened secretory epithelium, the clitellum (clitell = saddle), girdles the body
near the anterior end. The clitellum secretes a mucous cocoon, into which gametes and albumen
are released and where fertilization occurs.
Segments
The body is segmented and each segment is separated from its neighbors by a distinct
circumferential groove. The anteriormost true segment is the peristomium (peri = around, stome =
mouth; Fig 1, 13-59A). Anterior to the peristomium is a small dorsal lobe, the prostomium (pro =
before) that, for embryological reasons (it is not a product of the teloblast cells), is not considered to
be a segment. The peristomium encircles the large, ventral mouth and is an almost complete ring
around the body (Figs 2, 4, 13-59A). The prostomium fits into a small, dorsal notch in the
peristomium. The prostomium is a small lobe dorsal to the mouth and anterior to the
peristomium. The segments are numbered anterior to posterior beginning with the peristomium.
Count the preclitellar segments in your specimen. ____________ How many segments
contribute to the clitellum? _____________ Compare your counts with those made by other
students. Does the number of segments in each region appear to be constant or variable?
____________
The segments posterior to the peristomium are complete rings, without notches, and extend
uniformly for the entire length of the worm. The posteriormost division of the body is thepygidium,
which encircles the anus at the posterior tip of the worm. Like the prostomium, and for the same
reasons, the pygidium is not considered to be a true segment.
Chaetae
The eight small chaetae on each segment (except the first) are usually visible with
adequate magnification (25X). Chaetae are small chitinous bristles emerging from pores in the
integument on the ventral half of the worm. The chaetae are arranged in four pairs, two on each
side (Fig 6, 13-61A). Each segment has a pair of lateral chaetae and a pair of ventral
chaetae on each side. The chaetae are used as anchors when burrowing to hold parts of the
worm against the so that elongation of the animal results in controlled, usually forward, motion.
The chaetae are equipped with protractor and retractor muscles and are retractile. In your
specimen they may be withdrawn in some, or even all, of the segments. Careful examination,
however, should reveal some segments with extended chaetae. Chaetae are amber or brown,
slightly curved, and short.
Because the chaetae are sometimes used as landmarks to describe the position of other
structures, it is useful that they be given names. The four chaetae of each side of the segment are
designated with the names chaeta a, chaeta b, chaeta c, and chaeta d moving from ventral to
dorsal (Fig 6).
Figure 2. A ventral view of Lumbricus. The worm has been rolled slightly to reveal part of
the right side. Oligo55L.gif
 >1a. Rinse an unanesthetized worm with tap water and hold it in your hand on the stage of
the dissecting microscope. Focus on the lateral body wall and watch the chaetae. You may see
the animal retract or protract some of its chaetae while you watch. <
>1b . Run your fingers along the ventral and lateral surfaces of your anesthetized specimen
from anterior to posterior and then from posterior to anterior. Try to feel the chaetae and determine
if they point anteriorly or posteriorly. <
Pores
A little dorsal and anterior to chaeta b on each side of most segments, is a small opening,
the nephridiopore, near the anterior edge of the segment (Fig 2). This is the external opening of
a metanephridium, or kidney, a pair of which being present in most segments. The nephridiopores
are closed by sphincters and are very difficult to see when closed.
Earthworms are simultaneous hermaphrodites and each individual has complete female and
male reproductive systems including separate external gonopores. With the exception of the male
gonopores, these openings are small and difficult to find. They are best demonstrated with a bent
microneedle, which can be used to probe the surface of the cuticle until it slips into the previously
invisible opening.
On the ventral surface of segment 15 are conspicuous swellings associated with the
openings of the male vas deferens. The openings themselves are more difficult to see but each is
flanked by two low, glandular, transverse ridges, or lips, that are easily seen. The openings are
the male gonopores (Fig 2, 13-9A). During copulation sperm from the testes exit the body via
these pores. A shallow, inconspicuous sperm groove extends posteriorly from each male
gonopore to the clitellum and lies immediately lateral to chaeta b.
The two female gonopores, on segment 14 lateral to chaeta b, are the openings of the
oviducts. These can best be demonstrated by probing with a microneedle directed
anteriorly. Sometimes they are easy to see and sometimes not. Eggs from the ovary are released
from these pores.
The two pairs of seminal receptacles belonging to the female system are located in
segments 9 and 10 and their ducts open laterally in the grooves between segments 9-10 and 10-11
respectively (Fig 2, 13-59A). The four seminal receptacle pores may be difficult to find. Look for
them beside chaetae c and d.
All segments posterior to 12 bear a tiny coelomic pore located on the dorsal midline in the
groove between adjacent segments. These are almost impossible to see but can be demonstrated
relatively easily with the microneedle. They are often easier to see along the flatter posterior
portion of the worm. They open into the coelom and are used to leak coelomic fluid onto the
surface of the animal to keep it moist. The body surface is the respiratory surface and it must be
moist. Each pore has a sphincter to prevent unnecessary fluid loss.

Internal Anatomy
! Remove the worm from the dissecting pan and, while holding it in one hand, use your
finest scissors to make a shallow, dorsal longitudinal incision a little to one side of the dorsal
midline. Start by pinching the body wall with forceps and then cut through the pinch with the
scissors. This initial cut should begin immediately anterior to the clitellum and initially should
extend anteriorly for 2-3 segments. Be very careful that you do not cut the bright red
middorsal blood vessel that can be seen through the body wall on the midline.
>1c. Press the incision against a glass slide to transfer some of the coelomic fluid to the
slide then return the worm to the dissecting pan. To the slide, add a drop of 0.05% aqueous
methylene blue and a coverslip. Use the compound microscope to study the coelomic corpuscles,
or coelomocytes. Many are amoeboid and bear numerous short, narrow filopods, which move
very slowly, whereas others have no pseudopods. Some of the cells in the coelom are free
chlorogogen cells called eleocytes. <
Extend the dorsal incision anteriorly from the clitellum to the prostomium and posteriorly to
about segment 30-40. Be sure it is a little to one side of the dorsal midline. Reflect the body wall
and pin it to the wax with #1 insect pins as you go. Be sure the worm is in a convenient position
on the dissecting pan before you pin it. Plan ahead so that you will be able to observe any part of
the worm with the microscope after it is pinned. Insert the pins at 45 ° angles so you have plenty
of working room above the worm. It is impossible to conduct the dissection if the pins are inserted
vertically. Be sure the worm is completely immersed in 5-7% ethanol.
The body cavity, or coelom, is partitioned by septa, which are transverse sheets of thin
tissue that extend from the body wall to the gut tube. You will have to cut many septa (carefully,
with fine scissors) before you can move the body wall aside. Be careful that you do not cut any of
the large red blood vessels although it is inevitable that you will cut a few of the smaller vessels.
Body Wall and Coelom
Look at the cut edge of the body wall and, with magnification, identify its major
layers. Outermost is the thin, tough, iridescent cuticle composed of collagen fibers secreted by
theepidermis beneath it (Figs 3, 6, 13-59B). The epidermis is a monolayered columnar epithelium
containing numerous sensory and secretory cells. A subepithelial nerve plexus lies below the
epidermis.
A thick and conspicuous layer of circular muscle lies inside the epidermis. In fresh
preparations the circular muscle is reddish brown. The circular muscle fibers are divided into rings
by the circumferential grooves between segments.
Inside the circular muscle layer is a thick layer of white or gray longitudinal
muscle. Unlike the circular muscles, the longitudinal fibers run without interruption across
segmental boundaries. While it is not obvious in gross dissection, the longitudinal muscle mass is
divided into several bundles that run the length of the animal. You may be able to see the
divisions between these bundles on the inside of the body wall.
The innermost layer of the body wall is the somatic peritoneum. This is a thin, glistening
layer of squamous epithelium that covers the inner surface of the longitudinal muscles. A similar
layer of splanchnic peritoneum covers the surface of the visceral organs.
Figure 3. The anterior end of a dissected earthworm. The nephridia, except for those of
segment 7, have been omitted from the drawing. Oligo56La.gif
 The open space enclosed by the two peritoneal layers is the body cavity, or coelom. It is
divided into segmental coelomic compartments by double, vertical, transverse sheets of
peritoneum, muscle, and connective tissue called septa. Septa are thin and transparent. Most of
the septa were cut as you opened the body cavity but they may require additional cutting to reveal
organs of interest. The septa are penetrated by the gut, hemal system, and nervous
system. Longitudinal double sheets of peritoneum extend from the body wall to the gut tube and
aremesenteries. The gut tube is suspended by the dorsal and ventral mesenteries.
In many areas the splanchnic peritoneum is specialized to form bright yellow or
orange chlorogogen tissue (Figs 3, 6). Chlorogogen is involved in glycogen and lipid synthesis
and storage, amino acid deamification, and the synthesis of ammonia and urea. It has a high iron
concentration and may be the site of hemoglobin synthesis or breakdown. The color is due the
lipid stored in it. Chlorogogen is derived from splanchnic peritoneum, which explains its position
and distribution on the surface of the gut.

Chaetae
 The four pairs of chaetae of each segment can be seen internally in four longitudinal rows
along the floor of the coelomic cavity (Fig 3). The proximal heads of the chaetae are visible, with
high magnification, as tiny paired black specks. Each pair is in a chaetal sac, which secretes the
chaetae, and is attached to a set of tiny retractor and protractor muscles (Fig 13-59B).
The ventral chaetae of segment 26 are genital chaetae that help hold copulating worms
together. Chaetae in some segments associated with reproduction may be reduced in size and the
associated tissue modified for secretion.
Digestive System
Surface Features
Study the digestive system without damaging the hemal system. The gut is a straight tube
extending from mouth to anus. It is regionally specialized to perform the several functions,
including food procurement, storage, trituration, hydrolysis, absorption, reclamation, and feces
storage and formation typical of complete digestive systems.
The annelid gut is composed of foregut, midgut, and hindgut of which the foregut and
hindgut are ectodermal derivatives and lined with cuticle. The endodermal midgut does not
secrete a cuticle. The mouth opens into the short, thin-walled buccal cavity in segments 1-3 (Fig
3, 4, 13-61B, 13-63A).
The pharynx is posterior to the buccal cavity (Figs 3, 4) in segments 3-5. The pharyngeal
wall is thick and muscular. Numerous small radial muscles run from the pharynx to the body
wall. Contraction of these muscles dilates the highly distensible pharynx.
Take note of, but avoid damaging, the conspicuous, white cerebral ganglia (brain) located
atop the junction ofthe buccal cavity and the pharynx.
Posterior to the pharynx, the gut narrows to become the thin-walled esophagus in
segments 6-12 (Figs 3, 4, 13-61B). The posterior end of the esophagus is hidden by six large,
creamy white seminal vesicles that arch over it. Follow the esophagus through this region and
note that it is not a simple tube as it is anteriorly.
Figure 4. The anterior end of a dissected earthworm viewed from the dorsum. The blood
vessels, septa, and nephridia have been removed to reveal the gut and reproductive
system. Oligo57La.gif
 In about segments 10-11, the esophagus expands to become the wider esophageal
pouches (Figs 3, 4). Each pouch is marked with fine stripes but is hidden under the seminal
vesicles and septa.
Immediately posterior to the esophagus and esophageal pouches are two pairs of smaller
expansions of the gut, the calciferous glands (or esophageal glands). These bulge laterally from
the esophagus into the coelom and are usually white with an abundance of fine parallel red blood
vessels They are hidden by the third seminal vesicle and are immediately posterior to the fifth
heart. They remove excess calcium and carbon dioxide from the blood and secrete white crystals
of calcium carbonate into the gut. Calciferous glands play a role in the regulation of calcium
concentration and blood pH.
Posterior to the last pair of calciferous glands, the esophagus narrows again and then joins
the large, bulbous, thin-walled crop in about segment 12 (Figs 3, 4, 13-61B). The crop extends
posteriorly through several segments and is a food storage organ.
 Posterior to the crop the gut becomes the gizzard (Figs 3, 4, 13-61B). This large
expansion of the gut tube has thick, heavy, muscular walls and is the major site of mechanical
digestion. In it food is ground (triturated) into smaller fragments. Its muscles have a rich blood
supply and its transverse, slightly oblique, folds are conspicuous. The buccal cavity, pharynx,
esophagus, crop, and gizzard constitute the foregut.
The gut narrows again posterior to the gizzard and becomes the intestine (or midgut, or
stomach-intestine), which is the region for chemical digestion (hydrolysis) and absorption. The
intestine is heavily invested with yellow chlorogogen tissue.
The intestine extends from the end of the gizzard almost to the anus and its dorsal wall is
invaginated to form a typhlosole to increase surface area (Fig 6, 13-61A). The crease of the
typhlosole contains chlorogogen and connective tissue. The typhlosole is not visible in this view.
The anterior intestine, or saccular intestine, is specialized for synthesis and secretion of
enzymes and hydrolysis of food molecules. This region has thin walls that tend to form
blisters. The remainder of the intestine is absorptive and has thicker walls.
The extreme posterior end of the gut is the rectum, or hindgut. It lacks a typhlosole and
opens to the exterior via the anus. It is not visible at present.
It is not feasible to open the gut until after you have completed your study of the other organ
systems. Postpone study of the gut lumen and return to it after you have finished the reproductive
system.
Hemal System
The earthworm hemal system contains bright red hemoglobin which makes it very easy to
study in living specimens. The pigment is in solution rather than being cellular.
In preview, the basic plan of the system includes two major longitudinal vessels, one dorsal
and the other ventral to the gut (Fig 6, 3, 13-2, 13-61A). The two vessels are connected in each
segment by paired segmental vessels that supply and drain the tissues of the segment. The
dorsal vessel and five pairs of segmental vessels in the region of the esophagus are contractile and
function as hearts.
Blood moves anteriorly in the dorsal vessel and is pumped to the ventral vessel by the ten
segmental hearts. The blood then flows posteriorly in the ventral blood vessel. It leaves the
ventral vessel via the segmental vessels of each segment, goes to the organs and then returns to
the dorsal blood vessel.
The nervous system is provided with its own system of longitudinal blood vessels supplied
by vessels from the ventral blood vessel.
Find the large dorsal blood vessel on the midline of the dorsal surface of the gut (Fig 3,
13-61B). Peristaltic waves of contractions of circular muscles in the dorsal vessel move the blood
anteriorly. Watch the vessel as it contracts and see if you can tell which way the blood is
flowing. One-way valves in this vessel prevent backflow. Sometimes these valves can be seen
through the transparent walls of the dorsal vessel in the vicinity of the crop and gizzard.
Using 10-15X magnification, look ventral to the gizzard at the ventral blood vessel. This
vessel is attached to the ventral side of the gut by the narrow ventral mesentery (Fig 6, 13-2).
Follow the dorsal vessel anteriorly to the esophagus and find the five pairs of large,
contractile, segmental blood vessels which are the hearts (Fig 3, 13-61B). They encircle the
esophagus and pump blood ventrally, which is the reverse of the other segmental vessels. Trace
one of them from the dorsal vessel to the ventral vessel. If the hearts are beating, try to verify the
direction of blood flow. The hearts may be hidden by the septa and nephridia of their
segments. Remove these as necessary to reveal the hearts. Be careful you do not tear or cut the
hearts. Should you do so, the hemal system will bleed out and the hearts and vessels will be
much more difficult to see.
Note the numerous branches of the dorsal and ventral blood vessels in the region of the
crop and gizzard. The blood supply to the musculature of the gizzard is especially
impressive. Note also the blood vessels to the body wall musculature, septa, nephridia, and
seminal vesicles. These vessels are easy to see so long as they contain blood.
! Remove the hearts, dorsal blood vessel, nephridia, and septa from the area over the
esophagus and reexamine that region of the gut, locating any organs you could not find earlier (Fig
4).
 Respiratory System
Earthworms have no specialized respiratory structures and gas exchange takes place
across the general epidermis which can be moistened when necessary with fluid from the coelomic
pores. The integument is heavily vascularized to serve its respiratory function.
Nervous System
The earthworm nervous system is typically annelidan, consisting of a dorsal, anterior
brain, circumpharyngeal connectives, ventral subpharyngeal ganglion and a ventral nerve cord with
segmental ganglia, commissures, connectives, and segmental nerves (Fig 13-61-B, 13-62). The
brain is farther posterior in oligochaetes than in polychaetes and is no longer in the prostomium.
The brain, consists of a pair of cerebral ganglia atop the anterior pharynx in segment 3
(Fig 3). If your previous incision exposed the buccal cavity, the brain can be seen without further
dissection. Each ganglion is white and pyriform (pyri = pear, form = shape) and the two are
connected across the midline by a short transverse cerebral commissure. The brain is the
primary center for coordination of sensory and motor functions. It innervates the prostomium by a
pair of prostomial nerves which can be seen on the dorso-lateral surface of the buccal cavity.
A large circumpharyngeal connective exits the side of each cerebral ganglion and runs
ventrally around the pharynx to join the subpharyngeal ganglion ventral to the pharynx (Fig 13-
62). Carefully cut away the tissue on one side of the pharynx to expose the connectives and the
subpharyngeal ganglion. This ganglion serves the anterior three segments, including the
peristomium, and is a major center for motor control.
The double, solid ventral nerve cord characteristic of annelids exits the subpharyngeal
ganglion and extends posteriorly, on the ventral midline, for the length of the worm (Fig 5). The
nerve cord swells in each segment to form a segmental ganglion from which arise three pairs
of segmental nerves to nearby tissues (Fig 5). The segmental nerves are small and difficult to
see.
There are no special sense organs in earthworms but the body surface, especially that of
the head, bears receptor cells for taste, touch, light, and apparently vibration. The dorsum of the
body is more sensitive to light than the venter. Unicellular photoreceptors are present in the
epidermis and are most abundant on the prostomium. These photoreceptors are microscopic and
cannot be seen in gross dissection.
Excretory System
Almost all segments of the earthworm possess a pair of complex metanephridia (Figs 3, 6,
13-65B). In fresh specimens these are easily seen on both sides of each segment. They are
large, pale, and thin-walled. Each consists of a ciliated funnel, or nephrostome (nephr = kidney,
stome = mouth), which opens from the coelomic cavity into a long, tortuously coiled tubule that
penetrates the posterior septum of the segment and opens to the outside via a nephridiopore in the
wall of the adjacent segment (Fig 13-61A, 13-65B). The wide distal region of the tubule functions
as a bladder.
Reproductive System
Oligochaetes are simultaneous hermaphrodites and each individual contains complete and
simultaneously functional male and female systems. The female system produces eggs and
receives and stores sperm from the partner. The male system produces sperm and delivers it to a
partner during copulation. The reproductive system is restricted to a few preclitellar segments (9-
15). Most segments do not contain reproductive structures.
! Expose the reproductive system as follows. Free the anterior digestive system from
pharynx to gizzard from its septal and circulatory connections with the body wall but leave the gut
intact. Be careful that you do not harm the reproductive system in segments 9-15. Move the
digestive tube aside and pin it out of the way, but do not cut or remove it. Some of the
reproductive system is difficult to find but structures in bold type are easily identified.
Figure 5. Dorsal dissection of segments 9-16 of Lumbricus. The second right seminal
receptacle has been removed. The sperm reservoirs are drawn as if transparent so the
 testes and sperm funnels are visible. Oligo58L.gif

Male System
Look at the organs remaining in segments 9-15 (Figs 3, 4, 5, 13-61B, 13-67). Largest and
most conspicuous are the three pairs of irregular, yellowish seminal vesicles belonging to the
male system. Seminal vesicles store autosperm. (Autosperm is sperm produced by this worm.)
The posterior pair is by far the largest and the anterior pair the smallest. These specialized
coelomic spaces are the sites of maturation of sperm that originate in the tiny testes. The three
pairs of vesicles are lateral outpockets of two large, median sperm reservoirs in segments 10 and
11 (Fig 5). The small, inconspicuous testes are in the reservoirs. The male gonoducts (sperm
ducts, vasa deferentia) extend from sperm funnels in the reservoirs to the male gonopores on
segment 15.
Female System
Two pairs of small, spherical seminal receptacles are located in segments 9 and
10. These resemble the seminal vesicles but are smaller and more regular in shape.
Whereas the seminal vesicles are parts of the male system, the seminal receptacles
belong to the female system. The vesicles are the sites of maturation of autosperm. The
receptacles, in contrast, are the sites of storage of allosperm. Allosperm is sperm produced by, and
received from, the copulatory partner of this worm. Short ducts penetrate the body wall and
connect the seminal receptacles with external pores between segments 9-10 and 10-11.
A pair of inconspicuous ovaries is located in the anterior septum of segment 13 (Fig 5, 13-
61B, 13-67). The egg funnel is on the posterior septum of this segment and connects via the
oviduct with the female gonopore on segment 14.
>1d. Remove one of the seminal receptacles and place it in a drop of 0.75% saline solution
on a slide. Cut it in half and cover it with a coverslip. Examine the preparation with the compound
microscope. You may see masses of very active allosperm. Sperm are not always present in the
seminal receptacles. Bright white seminal receptacles usually contain sperm whereas pale cream-
colored ones frequently do not. <
 Study of the remaining features of the reproductive system is beyond the scope of this
exercise but its structures may be identified, at your discretion, with the help of Figures 5, 13-61B,
and 13-67 if desired. A pair of tiny ovaries in segment 13 release eggs into the coelom where they
mature. They are stores in a small pouch, the ovisac, which protrudes from segment 13 into
segment 14. A pair of female gonoducts, open from the coelom as the egg funnels, and transport
mature ova from the ovisacs to the female gonopores on segment 14.
Return now to the postponed study of the gut lumen and then come back to the cross
section slide below.
Gut Lumen
! Return the gut to its original position and open it from mouth to anterior intestine by
inserting the point of your fine scissors in the mouth and cutting posteriorly along the dorsal
midline. Pay attention to the structure of the gut wall as you cut. Where appropriate, observe the
contents and then remove them with jets of water from a pipet. The wall of the buccal cavity is
thin, as you already know. The walls of the pharynx are much thicker and have heavy longitudinal
folds that allow for expansion. The walls of the anterior esophagus are thin and unremarkable but
posteriorly the esophagus expands to form the esophageal pouches. Posterior to the pouches
are the two pairs of calciferous glands filled with calcium carbonate particles.
Open the crop. What is its function? What do you find inside it? The walls of the crop
are thin and folded longitudinally to permit expansion. The gizzard has very thick muscular
walls. Note the thick cuticular lining of the gizzard. If you carefully adjust the light, you will see
that the cuticle is iridescent like the epidermal cuticle. The cuticle can be grasped with a fine
forceps and stripped away from the underlying muscle and epithelium. Remove the cuticle and
look more closely at the soft tissue beneath it. The lining is grooved and ridged. You may see
several small stones in the gizzard. Their function is to assist in grinding the food.
Extend the incision posteriorly into the intestine but cut along the lateral wall rather than
the dorsal. The anterior intestine has very thin walls and the walls tend to form outward
blisters. This is the secretory saccular region of the intestine and is the site of chemical
digestion. The typhlosole is a large rounded fold of the dorsal gut wall that hangs down into the
gut lumen. In this region its walls bear conspicuous transverse folds (Fig 6).
Open the body wall all the way back to the anus. You need not be particularly careful with
this procedure. Moving posteriorly from the saccular region, open the intestine at various places
and find a region where the typhlosole is smooth and unfolded. The walls of the intestine are
thicker here. This region is chiefly absorptive. More posterior still is a region where there is no
typhlosole at all. This is the rectum. Its walls are a little thinner than those of the posterior
intestine but are thicker than the saccular region.
Remove the gut and set it aside. With the gut out of the way clear tissues away from the
ventral nerve cord to expose it to view. Trace it anteriorly to the brain.

Cross Section
>1e Study a commercially prepared slide of an earthworm cross section using the
compound microscope. The section is probably through the anterior intestine (Fig 6, 13-
61A). Look at the section first with low power (40X) and orient yourself. Find the body
wall, coelom, gut wall, and gut lumen (Fig 6).
Look at the body wall with high power (400X) beginning on the outside of the worm and
work inward. The outermost layer is the thin, noncellular cuticle (Fig 13-59B). The
cellularepidermis lies immediately below the cuticle and is a monolayered epithelium containing
abundant secretory and sensory cells. The secretory cells, which secrete the cuticle, are easily
recognized by their large open secretory vesicles. The very thin line at the base of the epidermis
is its basal lamina. A thin, inconspicuous, connective tissue dermis is situated just inside the
basal lamina and cannot be distinguished from it.
The next layer is the relatively thick circular muscle of the body wall. The muscle fibers of
this layer run perpendicular to the long axis of the worm and are seen here in longitudinal
section. Their nuclei are clearly visible.
 The thickest layer of the body wall is the longitudinal muscle layer, which lies inside the
circular muscle. Its fibers run parallel to the long axis of the worm and you see them here in cross
section. The featherlike appearance of these fibers when in cross section is distinctive. The
longitudinal muscle layer is divided into several bundles. A thin, squamous epithelium, thesomatic
peritoneum, covers the inner surface of the longitudinal muscles. The large open space enclosed
by the peritoneum is the coelom. Depending on the location of the section you may also see
metanephridia, chaetal sacs, and segmental blood vessels.
Look at the gut tube in the center of the coelom using 100X. The dorsal wall of the anterior
intestine is invaginated to form a large typhlosole which extends deep into the gut lumen. As a
result, the gut lumen is U-shaped, with the opening of the "U" directed dorsally. The thick layer of
cells bordering the lumen is the mucosal epithelium. Look closely at its free surface forcilia.
Figure 6. Cross section through the intestinal region of the earthworm, Lumbricus. The
section is slightly oblique. Oligo59La.gif

Moving from the mucosal epithelium toward the coelom there are, in order, connective
tissue, circular muscle, longitudinal muscle, and peritoneum but in the intestinal wall these layers
are poorly developed and probably cannot be distinguished from each other.
The epithelium of the foregut and hindgut secrete an extracellular cuticle that lines the
lumen and is best developed in the gizzard. The development and function of the epithelium and
of the muscle layers varies depending on the region of the gut.
Bordering the coelom, the outermost layer of the gut tube is the splanchnic peritoneum,
which on the intestine is specialized as chlorogogen tissue. The chlorogogen forms a thick layer
of large cells extending into the coelom from the peritoneum. Note that the space in the "U" of the
typhlosole is nearly filled with chlorogogen and connective tissue.
Dorsal to the gut tube is the large dorsal blood vessel. Ventral to the gut is the
smaller ventral blood vessel and large ventral nerve cord. The ventral vessel and the nerve
cord are suspended from the gut by the ventral mesentery. At the dorsal surface of the nerve
cord are three conspicuous giant axons.
A tiny longitudinal lateral neural vessel lies on each side of the nerve cord and a larger,
also longitudinal, subneural vessel lies ventral to it. The lateral neural vessels are supplied by
branches from the ventral blood vessel. They in turn supply the nerve cord with blood. The
subneural vessel drains the nerve cord. <

References
Brooks WK . 1890. Handbook of Invertebrate Zoology. Bradlee Whidden, Boston. 352p.
Brown A . (ed) 1950. Selected Invertebrate Types. Wiley, New York. 597p.
 Edwards CA, Lofty JR . 1977. The Biology of Earthworms. 2 nd ed. Chapman and Hall, London. 283p.
Freeman WH, Bracegirdle B . 1971. An Atlas of Invertebrate Structure. Heinemann, London. 129p.
Jamieson BGM ., 1992. Oligochaeta, Pp. 217-322 in Harrison FW, Gardiner SL (eds), Microscopic Anatomy of
Invertebrates 7, Annelida. Wiley-Liss, New York.
418 pp. Laverack MS. 1963. The Physiology of Earthworms. Pergamon,
Oxford . 206p.
Parker TJ, Haswell WA . 1951. A Textbook of Zoology, vol 1, 6 th ed. MacMillan, London.
Pearse V, Pearse J, Buchsbaum M, Buchsbaum R . 1987 . Living Invertebrates. Blackwell (Boxwood),
Palo Alto. 848p.
Rowett HGQ . 1957. Dissection Guides V. Invertebrates. Reinhart, New York. 56p.
Ruppert EE, Fox RS, Barnes RB. 2004. Invertebrate Zoology, A functional evolutionary approach, 7 th ed.
Brooks Cole Thomson, Belmont CA. 963 pp.

Supplies
Dissecting microscope
Compound microscope
Living nightcrawlers, Lumbricus terrestris
Worm-size dissecting pan (kippered herring tin with wax bottom)
#1 stainless steel insect pins
5-7% non-denatured ethyl alcohol in tapwater
Dissecting set with microdissecting tools
0.75% saline solution
0.05% aqueous methylene blue
Cross section slides of Lumbricus through intestinal region

Item Source
Live Lumbricus Local bait shop
Preserved Lumbricus Ward’s, Carolina, others
XS slides Triarch*, Wards, Carolina
Median Sagittal section, anterior end Triarch
Parasagittal section, anterior end Triarch, Ward’s
Near median sagittal section, anterior Ward’s
Nephridium wm Carolina, Ward’s, Triarch

*Triarch has a large variety of prepared Lumbricus slides