Laboratory Activity

Phylum Cnidaria
Supplies
1 living or preserved Aurelia
1 dissecting microscope
1 culture dish
1 planula wholemount slide
1 scyphistoma wholemount slide
1 strobila wholemount slide
1 ephyra wholemount slide
slides and coverslips
1 % toluidine blue in seawater
1 hypodermic syringe
1 % acetic acid

Introduction

The cnidarian body consists of a central blind sac, the coelenteron (= gastrovascular cavity), enclosed by a body
wall comprising two epithelia, the outer epidermis and the inner gastrodermis. A gelatinous connective tissue layer, the
mesoglea, lies between the two epithelia. The mouth opens at one end of the coelenteron and marks the oral end. The
mouth is at the tip of a process, the manubrium that elevates it above the oral surface. The opposite pole is the aboral end.
The imaginary line connecting the oral and aboral poles is the axis of symmetry around which the radial symmetry of
the body is organized. The mouth is usually surrounded by one or more circles of tentacles.
The defining cnidarian feature is, of course, their possession of stinging cells, or cnidocytes. Characteristic of the
epidermis, they are also sometimes found in the gastrodermis. Cnidocytes contain an explosive organelle, the cnida,
which, upon proper stimulation, inverts and ejects a slender, often barbed and toxic thread in the direction of prey or
predator. Three types of cnidae are found in cnidarians. Nematocysts (in nematocytes), spirocysts (in spirocytes), and
ptychocysts (in ptychocytes). All toxic cnidae are nematocysts whereas spirocysts are sticky, and the everted tubules of
ptychocysts are used for constructing feltlike tubes. Most cnidae are nematocysts and these are present in all three
higher cnidarian taxa. Spirocysts and ptychocysts are found only in Anthozoa.
The basic body plan described above can be manifest as a swimming medusa or attached polyp. In some taxa only
one generation is present whereas in others both are found. A life cycle featuring alternation of sexual, swimming
medusae with benthic asexual polyps is typical of many cnidarians.
All cnidarians are carnivores feeding on live prey which they usually capture using tentacles armed with
cnidocytes. Digestion occurs in the coelenteron which is typically equipped with ciliated canals for distribution of partly
digested food. Cnidarians are ammonotelic and diffusion across the body and tentacle surface eliminated the ammonia
from the body. Gas exchange is across the general body surface. The nervous system is a plexus of basiepithelial
neurons serving sensory and motor systems. Most cnidarians are gonochoric. The life cycle typically includes a planula
larva. Cnidarians are chiefly marine but the well-known Hydra is an exception.

Medusozoa
Medusozoa comprises those cnidarians whose life cycle includes a medusa generation that alternates with a polyp
generation. Symmetry is radial and tetramerous. Nematocysts are the only type of cnidocyte present. Included taxa are
Scyphozoa (jellyfishes) and Hydrozoa (hydroids, Hydra, Portuguese men of war, etc).
Scyphozoa
Scyphozoans, the jellyfishes, are cnidarians in which the medusoid generation is large and noticeable and the
polypoid is small and inconspicuous. The medusae, known as scyphomedusae, tend to be large, mobile, pelagic,
drifting, solitary carnivores and all are marine. The mesoglea of the medusa is thick, gelatinous, and accounts for most
of the mass of the organism, although it is mostly water. Cnidocytes are present, concentrated on tentacles, and are used
for prey capture and defense. Locomotion is by muscular contraction antagonized by elastic recoil of the mesoglea.
Sense organs are arrayed around the periphery of the organism. The sexes are separate. Polyps are small benthic
scyphistomae. Scyphozoa includes about 200 species in five taxa; Semaeostomae, Rhizostomeae, Cubomedusae,
Coronatae, and Stauromedusae.
Laboratory Exercise
Aurelia aurita , the moon jelly, is a common, cosmopolitan, largely coastal jellyfish. Place a living or preserved
specimen in a glass culture dish or pan of water. Use seawater if the animal is alive, tapwater if preserved. If possible,
select a small specimen that can be placed in a transparent petri dish and placed on the stage of the dissecting microscope
when desired.
External Anatomy
1) Examine the specimen, using the dissecting microscope as needed. The body consists of a large, relatively flat disc,
called the bell due to its bell-like shape (in life), or umbrella due to its umbrella-like shape, with the mouthparts hanging
below it. The lower, or oral, surface of the bell bears the mouth and is the subumbrella. The opposite, aboral, surface
is the exumbrella. The oral-aboral axis extends from the center of the aboral surface to the center of the oral
surface and is the axis of symmetry. The radial symmetry of cnidarians is organized around this axis. The margin of
the bell is shallowly indented at eight positions to impart a slightly scalloped appearance to its outline. Sensory
complices, the rhopalia, are located in the indentations between scallops.
Question a) Describe the organism in terms of its color, texture (with a gloved hand), and general appearance.
2) The outer surface is covered by a thin epithelium, the epidermis, which you cannot distinguish from the underlying
mesoglea. Deep within the animal, the coelenteron is lined with a second epithelium, the gastrodermis. Between these
two thin epithelial layers is the thick, jellylike mesoglea, which makes up almost all of the mass of the animal. The
mesoglea is a connective tissue composed of protein fibers and a watery ground substance and a few amoeboid cells.
Question b) Compare the cells of the two epithelium.
3) Orient the specimen in the dish with its oral surface uppermost, facing you. In the center of this surface are four long,
lacy oral arms of the manubrium. The oral arms are oriented at right angles to each other and form a conspicuous "X"
across the oral surface. The two axes that pass through the oral arms, perpendicular to the oral-aboral axis and to each
other, are the perradial axes. The axes bisecting the angles between adjacent oral arms are the interradial axes.
4) The surface of each oral arm bears two frilly, ribbonlike ridges that run its length and enclose a deep, ciliated brachial
groove. The grooves of the four oral arms meet at the mouth in the center of the oral surface. The margins of the
grooves bear small brachial tentacles. The four oral arms radiate outward from the short, inconspicuous vertical
column called the manubrium attached to the center of the oral surface.
Question c) Describe the way the organism take in and digest its food.
5) Inspect the margin of the bell and find the numerous, short marginal tentacles which bear cnidocytes. In
preservative the tentacles are greatly contracted but in life they are longer. There is an inconspicuous circular veil of
tissue, known as the velarium, central to the fringe of tentacles.
6) Make a wetmount of a tentacle and examine its nematocysts with high power of the compound microscope. You will
not be able to distinguish the cnidocytes themselves in these preparations but the oval nematocysts within them are
distinct. Look for the coiled thread within the nematocyst. If you have a living specimen, draw a little 1% acetic acid
beneath the coverslip and watch nematocysts discharge in response. Compare discharged and undischarged
nematocysts. (Acetic acid does not always cause the nematocysts to fire.)
7) Make a drawing of an undischarge and discharged nematocysts

Coelenteron
1) Aurelia feeds on zooplanktons which become entangled in the mucus on the aboral surface. The mucus is moved by
monociliated epidermal cells to the margin of the bell where it accumulates until removed by the tips of the oral arms.
Cilia in the brachial grooves then transport the mucus and food to the mouth.
2) The gut, or coelenteron (= gastrovascular cavity), of semaeostome scyphomedusae is an elaborate arrangement of
chambers and channels, lined by the ciliated gastrodermis and extending through the mesoglea and opening to the
exterior via the mouth. It is both the digestive system and the fluid transport system and is physically divided into two
regions to serve these two functions. Extracellular digestion occurs in a central set of chambers into which digestive
enzymes are secreted. From these chambers extend ciliated canals which distribute partly digested food to the body
where phagocytosis and intracellular digestion takes place.
3) The digestive region of the coelenteron consists of the stomach and gastric pouches. With your blunt probe, relocate
the mouth. It opens into a central stomach whose presence can be demonstrated with the probe gently inserted through
the mouth. Four semicircular gastric pouches open from the sides of the stomach and are the sites of extracellular
digestion. Each pouch contains a conspicuous horseshoe-shaped gonad on its floor. The opaque gonads are much
easier to see than the transparent gastric pouches but the edge of the gastric pouch is visible around the outside margins
of the gonad.
Question d) What is/are the reproductive methods used by the organism?
4) The passageway between the stomach and each gastric pouch is partitioned to form three short, side-by-side, ciliated
channels which move food between the stomach and the gastric pouch.
5) A fringe of short, threadlike gastric filaments is located on the floor of each gastric pouch along the inner margin of
each gonad. These secrete enzymes into the gastric pouches. With the probe, show that the filaments extend into the
pouches.
6) The distributive portion of the coelenteron consists of a set of narrow, ciliated canals that transport partially digested
food to and from the periphery. The system comprises a ring canal, in the margin of the bell, and a set of 16 radial
canals that extend from stomach to the ring canal. Numerous short blind canals arise from the ring canal and extend
into the marginal tentacles and sense organs.
7) Look carefully at the radial canals and note that some are branched and some are not. The canals lying on the perradial
and interradial axes are the four perradial and four interradial canals respectively. These canals are branched and
move materials from the ring canal back toward the stomach and gastric pouches. The perradial canals arise directly
from the stomach. The interradials and adradials arise from the gastric pouches. Between each perradial axis and its
neighboring interradial axis is an adradial canal. These eight canals are not branched and flow in them is away from
the gastric pouches.
8) If you have a living specimen, use a hypodermic syringe to inject toluidine blue/seawater or other dye into a gastric
pouch and watch it enter the radial canals. The outgoing ciliary current in the adradial canals will carry it to the ring
canal from which the incoming currents of the perradial and interradial canals will bring it back toward the pouch.
9) The septal funnels (= subgenital pits) are four invaginations in the oral surface of the bell, one below each gastric
pouch. The pits extend up into the mesoglea almost to the gonads and presumably provide the developing gametes with
oxygen. On cursory examination they seem to open into the gastric pouches but they do not. With a probe, (gently)
demonstrate that the pits are blind pouches and are not continuous with the gastric pouches.

Sensory Complex
10) The sense complices of Aurelia are the eight rhopalia evenly spaced around the margin of the bell. Each rhopalium
contains a statocyst for gravity detection, mechanoreceptors, chemoreceptors, and an ocellus for photoreception. Two
small, triangular, side by side rhopalial lappets are located beside the rhopalium. The complex is supplied by a tiny
branch of the ring canal.
11) Place the jellyfish in its dish with the oral side facing up on the stage of the dissecting microscope and find one of the
eight rhopalia on the margin of the bell. If your jellyfish is too large for the stage of the dissecting microscope, remove
a single rhopalium and the surrounding tissue and make a wetmount for study with the compound microscope. In the
center of the rhopalium is an elongate, ovoid, called the lithostyle. This structure is protected on the aboral side by a
semicircular flap of tissue called the rhopalial hood, which overhangs it. Below it are two triangular, side by side lobes
called rhopalial lappets. The lappets and hood extend slightly from the margin of the bell.
12) The lithostyle bears several sensory receptors, some more apparent than others. These include a pair of tiny dark
pigment cup ocelli, two obscure patches of chemosensory epithelium, and a conspicuous spherical statolith containing
tiny, opaque granules of calcium sulfate. The statolith is a device for the detection of gravity. A short branch of the
ring canal, the rhopalial canal, extends into the rhopalium to supply it with food. The canal branches into two lappet
canals and a central lithostylar canal. In living specimens injection of dye into the canal system may render these
canals more visible. <

Nervous System and Musculature

13) The nervous system and musculature are not visible in gross examination. Aurelia has a subepithelial nerve plexus
but no nerve ring. A concentration of neurons, called the rhopalial ganglion, is associated with each rhopalium.
14) The musculature includes a ring of circular epidermal muscles near the margin of the bell. Contractions result in
pulsations of the bell margin. This ejects water from the oral surface and propels the animal in the aboral direction.
Recovery is effected by elastic recoil of the mesoglea.
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 Laboratory Report
Protozoa
Name: _______________________________
Date: ____________________
Aurelia
Systematics:
Phylum: ___________________________
Class: _____________________________
Order: ____________________________
Labeling:
1) ___________________________
2) ___________________________
3) ___________________________
4) ___________________________
5) ___________________________
6) ___________________________
7) ___________________________
8) ___________________________
9) ___________________________
Hydra
Systematics:
Phylum: ___________________________
Class: _____________________________
Order: ____________________________
Labeling:
18) ___________________________
19) ___________________________
20) ___________________________
21) ___________________________
22) ___________________________
23) ___________________________
24) ___________________________
25) ___________________________
26) ___________________________
27) ___________________________
Year/Section: ____________
Grp No.:___________
Family: ___________________________
Genus: ___________________________
10) ___________________________
11) ___________________________
12) ___________________________
13) ___________________________
14) ___________________________
15) ___________________________
16) ___________________________
17) ___________________________
Family: ___________________________
Genus: ___________________________
28) ___________________________
29) ___________________________
30) ___________________________
31) ___________________________
32) ___________________________
33) ___________________________
34) ___________________________
35) ___________________________
36) ___________________________
Answer to Questions:

Question a) __________________________________________________________________________________
___________________________________________________________________________________________
__________________________________________________________________________________________

Question b) __________________________________________________________________________________
___________________________________________________________________________________________
__________________________________________________________________________________________

Question c) __________________________________________________________________________________
___________________________________________________________________________________________
__________________________________________________________________________________________

Question d) __________________________________________________________________________________
___________________________________________________________________________________________
__________________________________________________________________________________________

You may put your answers on another piece of bond paper.

1) Compare the tissue organization of Cnidarians and Poriferans.
2) Make a diagram showing the life cycle of Aurelia. (on another piece of paper)
3) Drawing for step number 7