Chapter 3: Phylum Cnidaria Classes Hydrozoa: Scyphozoa: Anthozoa: Hydra, Obelia, Gonionemius, Physalia Aurelia Metridium Class Hydrozoa Hydra Living Hydra is attached to the substrate by a basal disc. The stalk, or stem, of the Hydra is called the column, or peduncle. It begins at the basal disc and extends to the tentacles. A cone-shaped section rests on the peduncle. The tentacles extend from the base of this cone. The cone is called the hypostome, and is at its apex is the mouth. As you observe the peduncle you will notice that the middle of it is lighter than the periphery. This is due to the presence of a cylindrical cavity, which occupies the interior of Hydra. The cavity is called the gastrovascular cavity; the only entry or exit into the gastrovascular cavity is the mouth. Cnidocytes, or stinging cells, are the distinctive characteristic of Phylum Cnidaria Histology Take a slide that shows a longitudinal and cross-section of Hydra and study the histology. A cross section of the body should show two layers of cells. The external layer is the epidermis. It is one cell layer thick, and most of the cells are epitheliomuscular cells and appear rectangular. They have a nucleus bearing a conspicuous nucleolus. At the base of these cells are smaller, triangular cells with smaller nuclei. These are the interstitial cells. The other prominent cells in the epidermis are cnidocytes. The basal disc epidermis is basically of tall glandular cells. Their glandular nature can be recognized by the numerous eosinophilic (red) granules in their cytoplasm. Sensory cells are also present in the epidermis, but are not demonstrated well with hematoxylin and eosin. The surface of the epidermis is covered by a thin cuticle, which is very closely applied to the cell membranes. The basal portion of the cells rest on the mesoglea. In Hydra, it is very thin, appearing as no more than a bounding line. It is non-cellular. On the interior of the mesoglea is a second layer of cells, the gastrodermis. The two prominent types of cells in the gastrodermis are the digestive cells and the glandular cells. Both are large cells. The glandular cells are recognizable by 12 numerous eosinophilic globules, which are somewhat larger than those encountered in the basal disc epidermis. The digestive cells do not have these globules, but may have vacuoles, which are not stained. Both cells have a small number of flagella projecting into the lumen (gastrovascular cavity or coelenteron) of the organism. The flagella have been altered too much to be visible in the preparations you have. Interstitial cells and sensory cells are also present. Compare the external surface contour of the epidermis with the internal surface contour of the gastrodermis. Figure 2.1. Hydra cross-sections (A. female and B. male). Figure 2.2. Hydra whole mounts (A. male and B. female). 13 Reproduction 1. 2. Asexual by budding: Look at slides of total mounts with buds and longitudinal sections. After these buds attain a certain size, they develop a mouth, hypostome and tentacles, then drop off and form a basal disc. Sexual: Under certain stimuli the epidermal cells of Hydra develop regional thickenings, or gonads (testis or ovary). These appear as bumps on the side of the peduncle. The testis (Fig. 2.2A) tends to develop closer to the tentacles, while the ovary (Fig. 2.2B) develops more basally. Look at the slides of the cross sections of an ovary and testis. The testis results from interstitual cells undergoing meiosis and producing sperm. These germ cells and their precursors distinguish the testis as a structure made up of many very small cells. The ovary, by contrast, has the large ova, which are few in number, and adjacent somewhat smaller nurse cells, which are numerous. Distinguish between the two organs and their parts. Hydroid Stage The stage you have been studying is called the hydroid, or polyp stage. This is one of the two stages present in the life cycle of most cnidarians. The other stage is the medusoid stage (jellyfish). The polyp stage is found in all three classes of the phylum, but is very small in the class Scyphozoa. Many polyps, unlike Hydra and Metridium, are colonial. A common marine Hydrozoan colony is Obelia. Look at a slide of the Obelia hydroid (Fig. 2.3). There are two types of zooids (functional polyps) in a colony. The hydranth is the feeding polyp, and is shaped much like the Hydra possessing tentacles. The other zooid is reproductive, and is called the gonangium. On its stalk there are many red rosette structures. These are medusa buds , which are produced asexually and will eventually leave the colony and become free-swimming jellyfish. 14 Figure 2.3. Polyp form of an Obelia colony. Another colony form is Physalia (Portuguese man-of-war). It has an air bladder (pneumatophore) which functions for buoyancy. The long tentacles hanging from the base of the bladder are supplied with nematocysts. They are called dactylozooids and have the function of paralyzing prey. Immediately under the bladder are gastrozooids, which digest the prey. They are numerous. Gonophores (reproductive zooids) and jelly zooids are also present. Physalia is, in reality, a mixture of a hydroid and medusoid generation with some zooids, polypoid, and other medusoids. Observe the specimens on demonstration. Medusoid Stage The medusoid, or jellyfish, stage is topologically identical with the hydroid stage. The medusa differs from the hydroid in that it has more tentacles, is free swimming, and has a much thicker mesoglea. Look at the slide of the Obelia medusoid. This is a very small medusoid but it shows the convex surface (exumbrella) and subumbrella (concave) surface. The projection from the middle of the subumbrella surface is called the manubrium, and like the hypostome of the hydroid contains the mouth at the apex. Frequently, fixation of the specimen causes it to invert, making the manubrium project atypically from a convex surface like a handle. In an oral or aboral view, four canals can be seen radiating out from the center of the coelenteron. These are the radial canals, which communicate with the central gastrovascular cavity and a ring canal at the base of the tentacles. Single spherical objects are 15 associated with each radial canal. These are the gonads. The histology will not be drastically different from that of Hydra with the exception of the thick mesoglea. Obelia is too small for detailed study of one characteristic of the medusa of the class Hydrozoa, and instead the jellyfish Gonionemus (Fig. 2.4) is used. Gonionemus is structurally like the Obelia medusa, except that the broken ribbon of gonads associated with them obscures the radial canals. Also, in Gonionemus, one can recognize a bend in the tentacles near the end. This is an adhesive pad. Projecting in from the inner edge of the ring canal region is a shelf-like, membranous, structure called the velum. This is found only in Hydrozoan jellyfish. Hydrozoan jellyfish are usually less than an inch in diameter. Figure 2.4. Scyphozoan jellyfish Gonionemus. Class Scyphozoa Scyphozoan Jellyfish Anthozoans have no jellyfish stage. The large jellyfish, therefore, belong to the class Scyphozoa. They do not have a velum, contain their gonads internally, and possess sensory structures around the bell margin. Aurelia is an example of this class. Size has been mentioned is one way to distinguish medusae of this class (Fig. 2.5). The tentacles are short, and are arranged around the periphery, as in other medusae. The manubrium is very short, and 16 perhaps absent. From the mouth region there extend four long lobes called oral lobes, or oral arms. They reach to the bell margin. The gastrovascular cavity inside the mouth opens into it four large cavities, which when viewed together, resemble a four leaf clover. From these cavities, called gastric pouches, and between the gastric pouches radial canals go to the bell margin. Thus, there are many radial canals, not just four, as with the Hydrozoan medusa. On the oral surface of the gastric pouches are dark staining, horseshoe-shaped, gonads. Also on the oral surface of the gastric pouches are small projections called gastric filaments, which contain nematocysts. Around the bell margin are periodic indentations. Lobes called lappets flank them. Between each lappet are structures called rhopalia, which house a statocyst, or organ of balance, a photoreceptor, and a chemoreceptor. Figure 2.5. Scyphozoan medusa, Aurelia. Class Anthozoa Metridium senile (Sea anemone) This organism is also a hydroid. However, it is in the class Anthozoa. Look at a mature specimen on demonstration. Then, look at a cross-section. In a cross-section, two distinctions are capable of being made with similar crosssections of Hydra and these serve to separate the two classes. Metridium has the gastrovascular cavity divided into chambers by partitions called septa (Fig. 2.6). Some of the separations are complete, and are called primary septa. Or, the partitions may extend only a short distance into the cavity, and are secondary and tertiary septa. Secondly, the mesoglea is greater in extent 17 than in Hydra. Locate the muscle bands in the larger septa. Locate the septa, ostia, and acontia on the clay model. Figure 2.6. A. Diagram of a sea anemone. B. Cross-section through an anemone. Radial Symmetry Throughout this exercise there has been no use of the orientation terms used with organisms possessing bilateral symmetry. It is not meaningful to speak of the dorsal or ventral surface of a hydroid or a medusoid. This is because the organisms are radially symmetrical. Several planes can be passed through the organism, which will divide it into mirror image halves. Replacing the terms used in bilaterally symmetrical animals are the terms oral surface, the surface on which the mouth is found, and aboral surface, the surface opposite to the one on which the mouth is found. The main axis is the oral-aboral axis. Movement away from this axis is peripheral. In this course you will study three phyla that have radial symmetry. Phylum Review Phylum Cnidaria (means "nettle-bearing") sac-like body gastrovascular cavity lined by gastrodermis used for extracellular digestion single orifice for mouth and anus most have tentacles around oral end 18 nerve net; no central nervous system polymorphism; polyps and medusoid stages radial symmetry nematocysts in cnidocytes o Class Hydrozoa (means "water serpent animal") Hydra, Gonionemus, Obelia, Physalia solitary or colonial asexual polyps and sexual medusae--some may have either polyp or medusa stage reduced or absent hydranths without mesentaries medusae with a velum aseptate gastrovascular cavity ectodermal gonads (when present) o Class Scyphozoa (means "cup animal") Aurelia includes most large jellyfish solitary polyp stage reduced or absent bell-shaped medusae without velum microscopic septa divide gastrovascular cavity tetramerous radial symmetry endodermal gonads mesoglea enlarged o Class Anthozoa (means "flower animal") Metridium (sea anemone) corals (both soft corals and hard corals) solitary or colonial only polyp forms; medusa stage absent gastrovascular cavity subdivided by mesentaries (septa) bearing nematocysts usually either hexamerous or octamerous radial symmetry endodermal gonads mesoglea enlarged to form collenchyma (true mesoderm) Subclass Hexacorallia (means “divided into six”) sea anemones and hard corals hexamerous mesenteries in pairs Order Actiniaria 19 o sea anemones o well developed pharynx and siphonoglyph Order Scleratinia (means “hard”) o hard corals o calcareous exoskeletons Subclass Ceriantipatharia tube anemones and thorny corals mesenteries unpaired Subclass Octocorallia (means “divided into eight”) sea fans, soft corals, sea pansies octomerous symmetry soenia Phylum Ctenophora Ctenophores are structurally similar to Cnidarians in that they have two embryonic germ layers and radial symmetry. They contain rows of comb-like plates used for locomotion. Many are also luminescent and glow a bright green color when disturbed. A jarred specimen is available for you to examine. 20 Chapter 4: Phylum Platyhelminthes Classes Turbellaria: Planaria, Dugesia dorotocephala Trematoda: Conspicuum icteridorum, Clonorchis (flukes) Cestoda: Taenia pisiformis (tapeworm) Class Turbellaria Planaria Whole Mount Cilia, confined to the ventral side, provide locomotion for the organism. Near the anterior end are 2 dorsal eyespots (ocelli) and lateral, triangular auricles. The eyespots are collections of photoreceptors. The pigmented area you see is merely a light absorbent area toward which the ends of the neurons are pointed. As with most light receptors, the ends of the nerve cells are pointed away from the incoming light. Are the ends that are stimulated dendrites or axons? Review the neuron if you do not know. The auricles contain chemoreceptors and thigmoreceptors. When planarians feed, the protrusible pharynx projects out the mid-ventral mouth. Note the darkly colored, highly branched intestinal caeca that can be observed through the translucent body wall. Prepared slides are available to you. On each slide is a whole mount of a planarian, which has been fed a black pigment and a second planarian that has not been fed the pigment. The slide is thickly mounted, and you should not try to use the high power of the microscope. The organism, which has been fed pigmented material, has its digestive system thoroughly stained. You can see the pharynx and the three branches of the intestinal caeca. The branches have smaller branches, and thus you have illustrated the dendritic (branched) nature of the digestive system (Fig 4.1). Many of the organ units of platyhelminthes have a dendritic pattern. Platyhelminthes lack an anus. The digestive unit is thus referred to as an incomplete digestive system. Most metazoans have a complete digestive system. Think of the problems that accompany an incomplete digestive system. 21 Figure 4.1. Planarian whole mounts. Planaria Cross-Sections Obtain a slide of three cross-sections through a planarian. Look at the three cross-sections with a microscope. From what you have learned about the digestive unit you should be able to distinguish the sections as anterior, mid-body and posterior. Go to a higher power. You should be able to distinguish cilia on one surface and thus locate the ventral side. There is a single layer of cells, which lines the outside of the body. These cells bear cilia on the ventral side, and dark staining rhabdites on both sides. The epidermis also produces glands, which have a reddish secretion. These glands are larger at the edges of the organism, and there project into the body. Medial to the epidermis is a muscle layer. It has two parts: an external circular layer, and an internal longitudinal layer. There are also transverse muscle fibers that allow for more varied movement. Virtually all the interior tissue is made of parenchyma cells, digestive cells, and muscle. The muscle is present in thin strips oriented dorsoventrally, or transversely. The large digestive cells surround the only cavities you see in cross-section. Nervous and excretory tissue is present, but not easy to resolve. The remainder is thus parenchymal tissue. Parenchyma introduces you to connective tissue. All of the cells between the organs are parenchymal cells. They serve as a packing tissue, but they are also important in regeneration. Parenchymal cells are totipotent cells. That is, they have the capacity under appropriate stimulus to develop into any other kind of cell in the body. Most metazoans lack cells with this potential. Intriguing possibilities would exist if we could acquire that potential in vertebrates. Regeneration in planarians is possible because of the 22 totipotent nature of the cells and because of sufficient nutrition being available while the digestive unit is regenerating. Class Trematoda Digestive system Obtain a prepared slide of Conspicuum icteridorum, the gall bladder fluke of the common grackle (Fig. 4.2). Label the parts as you find them in looking through the dissecting microscope. The digestive system is much like that of planarians, but with several modifications. First, the mouth is located at the anterior end and is surrounded by a muscular oral sucker. Leading posteriorly from the oral sucker is a tube like esophagus, which branches into the two intestinal caeca that run almost to the posterior end of the organism. The entire digestive unit resembles an inverted ‘Y’ with a short basal stem. Around the esophagus and immediately posterior to the oral sucker is the muscular spherical pharynx. The other non-reproductive system that is somewhat visible is the excretory system. Excretory system At the posterior end is the excretory pore, which drains the large excretory bladder. The excretory bladder will be visible as a central clear area, since the walls of the organ are very thin. Two excretory ducts continue forward, branch and eventually end as flame cells. Flame cells are only seen on living specimens. Posterior to the bifurcation of the intestinal caeca is a ventral sucker (acetabulum). This structure attaches the symbiont to the wall of the gall bladder. It contains no opening into the interior; it is only a holdfast organ. Reproductive system The male reproductive system begins with two dark staining round or lobed testes just posterior to the acetabulum. From the testes, ducts (vasa efferentia) go anteriorly to form a single vas deferens. The short vas deferens ends in a large elongate cirrus sac, which opens via the male pore into the genital atrium. Inside the cirrus sac is a basal enlargement, the seminal vesicle, which narrows to form the cirrus. The cirrus is used to transfer sperm to another worm by being projected out the male pore, genital atrium and common genital pore and into the genital atrium of a second worm. Overview of male reproductive flow: 23 testes vas efferens pore vas deferens seminal vesicle cirrus genital atrium common genital pore male genital The female reproductive system begins with two unequally stained bodies in the center of the organism. The darker of the two bodies is the ovary, and the lighter the seminal receptacle. Both bodies are around a region called the oötype. They have ducts connected to the oötype. The oötype cannot be seen well, except in sections, but it is a very important crossroads and activity center in the female reproductive system. We will discuss it in more detail in lecture. The uterus arises from the oötype and the vitellarian ducts empty into the oötype. The uterus obscures most of the organs. It is a coiled structure, filled with many darkly staining eggs. After it leaves the oötype, the uterus goes posteriorly and then anteriorly to open into the genital atrium through the female pore. Lateral of the intestinal caeca are the vitellaria (yolk glands), which drain into the vitellarian ducts. Overview of female reproductive flow: seminal receptacle ® seminal receptacle duct ¯ ovary ® oviduct ® ootype ® uterus ® female genital pore ® ­ genital atrium ® common genital pore vitellaria ® vitellarian ducts 24 Figure 4.2. Diagram of a typical Conspicuum including reproductive and digestive systems. Class Cestoda Obtain a prepared slide of Taenia pisiformis, one of the dog tapeworms (Fig. 4.3). Do not use high power in observing this specimen. On the slide are representative sections of one tapeworm. One unique feature is the absence of a digestive system. Since the organisms are symbionts in the intestine of vertebrates, they have come to depend on the host to provide the monomeric nutrients. The tapeworms have four basic regions. Anteriorly is the first region: the scolex (head), which in this case contains four suckers, and an anterior protrusion called a rostellum, which contains hooks. The hooks and suckers provide a means of attachment. Posterior to the enlarged scolex is the second basic region: the thin, unsegmented neck. Though seemingly inconsequential except as a connecting device it is the site of continuous asexual reproduction. The posterior end of the neck region continues to produce new segments, proglottids, throughout the life of the worm. The initial proglottids produced by the neck are small and undifferentiated. They are called immature proglottids, and are the third basic body region. 25 Little internal structure is visible. However, as they grow and differentiate each segment develops a reproductive system much like that of flukes. When the reproductive system is visible the proglottids are called mature proglottids, and are the fourth body region. Each mature segment is capable of mating. A consequence of this is the production of large numbers of eggs, which are frequently maintained in the proglottid. The segment becomes engorged as a large egg sac and is then called a gravid proglottid. Laterally in each segment is a longitudinal excretory duct. There are actually four of them and they drain numerous flame cells. Longitudinal nerve cords parallel the excretory ducts. The lateral genital pores frequently have the cirrus projecting from them. Overview of male reproductive flow: testes vas efferens pore vas deferens seminal vesicle cirrus genital atrium common genital pore male genital Overview of female reproductive flow: vitellaria ® vitellarian ducts ¯ ovary ® oviduct ® ootype ® uterus ­ shell gland Overview of flow of sperm into the worm: common genital pore genital atrium female genital pore vagina seminal receptacle seminal receptacle duct oötype 26 Figure 4.3. The four areas of a typical cestode, including a detailed mature proglottid. Pseudocoelomates Phylum Rotifera Since rotifers are unable to swim against water currents, they are considered to be planktonic. In contrast, organisms such as fish that can swim against water currents are classified as nektonic. You should be able to discern a hardened, segmented cuticle called a lorica (Fig. 4.4). Note the ciliated corona, which is composed of 2 trochal disks and in life, whorls around and gives the appearance of a wheel (hence the name of the phylum) connected to the anterior end of the organism. The other characteristics of rotifers will be best observed on whole mounts of rotifer slides. Check with your texts for verification of individual features. Identify the pharynx, or mastax, that contains two darkly staining jaws called trophi. In female rotifers, the ova and yolk are produced in germovitellaria, which are combined ovaries and yolk glands. The germovitellaria as well as intestine and excretory ducts empty directly into a cloaca that, in turn, empties to the outside via the anus. 27 Figure 4.4. Rotifer structures. Phylum Acanthocephala View the slides provided. Be able to identify the organism and know it is a pseudocoelomate. Phylum Review Phylum Platyhelminthes (means "flat worms") many dendritic organ systems parenchymal tissue composed of totipotent cells (thus have capability of complete regeneration of lost body parts) most primitive animals with excretory system acoelomate; lacks either eucoelom or pseudocoelom triploblastic (3 germ layers--ectoderm, mesoderm, endoderm) o Class Turbellaria (means "stir like") planaria (Dugesia); brown planaria we use is Dugesia dorotocephala free-living flatworms 28 integument (living, outer epidermis) ventral cilia incomplete digestive system since lacks anus; triradiate intestine with numerous caeca or diverticula o Class Trematoda (means "holes form") Conspicuum icteridorum, Clonorchis, Telorchis, Cephalogonimus flukes: both monogenetic (1 host in life cycle) and digenetic (2 hosts), sometimes trigenetic (3 hosts) tegument (nonliving, outer cuticle) lacking cilia incomplete digestive system; diradiate intestine endoparasitic lacks hooks; has oral sucker and ventral sucker (acetabulum) for attachment to host o Class Cestoda (means "girdle form") Taenia pisiformis tapeworms tegument lacking cilia no digestive system endoparasitic has anterior scolex with hooks for attachment to host body divided into series of proglottids Phylum Rotifera (means "wheel bearing") very small, planktonic marine, freshwater, terrestrial; epizoic, parasitic characteristic ciliated crown (corona) which looks like a wheel when beating shape correlates with mode of life Phylum Acanthocephala (means "spiny head") proboscis with recurved spines lacks digestive system flattened body all are endoparasitic lacunar system for nutrient travel 29