Examine your petri dishes of regenerating planarians. If some of you proved to be excellent surgeons than you can consider using planaria as your project. The goal here is to cut the planarians various ways and then speculate on the distribution of neoblasts that would explain your results. What other factors (genetics) could be important
General introduction
Don't you ever laugh as a hearse goes by,
For you may be the next to die.
They wrap you up in a big white sheet,
And cover you up from your head down to your feet.
They put you in a big black box,
And cover you up with dirt and rocks.
All goes well for about a week,
And then your coffin begins to leak.
The worms crawl in, the worms crawl out,
The worms play pinochle on your snout.
They eat your eyes, they eat your nose,
They eat the jelly between your toes.
A big green worm with rolling eyes,
Crawls in your stomach and out your eyes.
Your stomach turns a slimy green,
And pus pours out like whipping cream.
You spread it out on a slice of bread,
And that's what you eat when you are dead.
To watch animals feed you will need to place food in containers.
There is a danger of fouling the containers. Please be careful and add only the amounts specified in the instructions to your containers. If a container looks foul to you, call the lab instructors attention to it so she can add new water. If she instructs you to do so, make sure you know whether you are to add spring or salt water to your bowl.
A rule of thumb is that Clitellates (earthworms and relatives) are terrestrial or fresh water forms, Polychaetes are salt water forms and so are Sipunculids and Echiurans. We have plenty of the microscopic fresh water forms, so these can be dumped when you

are done. It will ensure that the rest of the population does not suffer.
Background:
Annelida consists of the segmented worms in the major classes Polychaeta
(bristleworms), and Oligochaeta (earthworms and relatives), with a total of about
12,000 known species in marine, freshwater, and terrestrial environments.
Most annelids have chitinous bristles, or chaetae, secreted by epidermal cells, that project from the body. The body wall consists of a collagenous cuticle secreted by the monolayered epidermis. The coelom is large, segmentally compartmented, lined and well developed in polychaetes and oligochaetes but reduced in leeches.
The circulatory system of most annelids is a set of tubular vessels, some of which are contractile and serve as hearts. The circulatory 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, circulatory, 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 clusters of germ cells were present in each segment and released developing gametes into the coelom.
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Polychaetes
Most segmented worms found in marine environments represent a major evolutionary branch of annelids--Class Polychaeta. The name means "many bristles." The bristles, or setae, project from side flaps called "parapodia." Parapods have a variety of shapes -- adaptations for different habitats and life-styles. Species living in rocky environments have leg-like parapods used in walking. In burrowing species, parapodia function as digging paddles. In some worms the parapodia are short and combine with peristaltic contractions to move the worm through the mud. In others, the parapodia contract following

their power stroke, streamlining the worm for passage through the mud.
Sedentary Polychaetes
Sedentary polychaetes are usually adapted to living permanently in tubes or burrows; some attach themselves to rocks or piers. Many sedentary polychaetes, like the lugworm, Arenicola , live in burrows in sand or mud. The majority, however, are tube builders. Tubes of different species vary greatly in their composition and structure. They may be composed of sand, shell, or other particles held together with mucus, or made entirely of organic substances secreted by the worm that harden on contact with water.
Sedentary polychaetes have greatly modified head regions for specialized feeding habits. Many are adapted for feeding on organic matter deposited on the ocean floor.. The parapodia are reduced in the sedentary polychaetes, and the setae of many tube-dwelling forms are hooked to help the worm hold itself to the wall of its tube.
We have living representatives of the tube dwelling polychaetes.
The Sabellids are commonly known as "fanworms" or "feather duster" worms due to a colorful appearance of a maroon or redcolored tentacular branchial crown . They are indirect depositfeeders. They have no proboscis. While feeding they spread out their branchial crowns to trap suspended particles and to sweep with their long mobile palps picking up deposited material (filter-feeding). Only suitable sized particles are ingested while others are rejected at the mouth. The more primitive sabellids members making temporary mucus tubes and creep around actively.. The large ones never leave their tubes and are associated with shallow water but smaller ones are able to move around. and are common in deep seas.

1. Obtain a feather duster worm and place in a dish of sea water
(Salt water only please).
Allow it to acclimatize to its new environment. Study its feeding device under the stereoscope.
A. Include a movie or several photographs of its filter feeding in your notebook.

Worm in tube
The particles that land on the grooves can be sorted with respect to size.
2. There may be other sedentary worms available for you to observe. Obtain photographs. and compare the morphology of at least one other specimen to the feather duster worms. Pay particular attention to the head, which will tell you how these

animals feed. For example, some sedentary worms have long tentacles with groves on them. These act as the “feathers” of a feather duster worm. The food particles are moved along by cilia in these grooves toward the mouth.
This week, another example of a sedentary worm we have in the laboratory is Spirorbis
Spirorbis
In this genus and in the family Spirorbidae all species live in permanent calcareous tubes cemented to algae, shells, or rocks.
These have cemented themselves to the walls and filter in the large
60-gallon tank.
Obtain a few specimens of this microscopic worm and place them in a small Petri dish so they can acclimatize to their new conditions. Be prepared to watch them for a few minutes so you can see them unfurl their tentacular crown .
Another common reef worm is Phyllochaetopterus.
Phyllochaetopterus individuals build a tube out of "hardened" mucus in which they cement sand grains. These tubes can be up to an inch or so in length and are about the diameter of a thin piece of pencil lead. They will be oriented vertically in the sediments or occasionally found filling pre-existing holes in rocks. The worms themselves are quite small, less than a quarter inch long; in fact, in most cases less

than a tenth of an inch. They are numerous in our reef tank, but difficult to remove without the worm escaping. So there are fewer individuals of these to observe than of the Spirobus.
Obtain a video if possible of Spirobis of Phyliochaetopterus
“feeding”.
3. Errant Polychaetes
Errant polychaetes include actively crawling or swimming forms, which may, however, also spend time in burrows or crevices, or under rocks on the seashore.. Many are predators on small invertebrates; some are scavengers.. Most errant polychaetes have well-developed head regions, which bear eyes, sensory tentacles, and a specialized organ, the nuchal organ, thought to detect chemicals. The anterior end of the gut often forms a protrusible structure, the proboscis, sometimes equipped with strong chitinous jaws and used in feeding. The setae of some polychaetes, e.g., the tropical fireworm, are composed of calcium carbonate rather than chitin and are hollow. These brittle setae are easily broken off and contain a toxin that produces a painful reaction in humans. In the scaleworms, a series of overlapping scales form a covering over the animal's upper surface. In the sea mouse these scales are completely covered by long, slender, feltlike setae projecting from the parapodia.

We only have one representative of an errant polychaete and that is the brstleworm. All you nee to do with the preserved specimens we have is to look at the jaws and compare these structures with the fans observed in your filter feeders.
Many errant polychaetes are ferocious predators and most aquarists remove them as soon as they are spotted. We may have some live fire or bristle worms available. Please wear gloves when handling these specimens.
They are called fire worms for a reason.
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Clitellata
Clitellata, the sister taxon of Polychaeta, includes earthworms and their allies. The head is reduced and its sensory functions minimized.. Obvious in most is a girdle-like band of secretory

epidermis, the clitellum, near the anterior end of the worm, a collar that forms a reproductive cocoon during part of their life cycle.
Oligochaetes are members of the Clitellata that lack the parapodia characteristic of polychaetes although setae may be present. Also lacking are head appendages These worms are hermaphroditic and have gonads and reproductive equipment restricted to a few specialized genital segments. Development is direct, without a larva.
In the laboratory, you will find several living representatives of
Clitellata annelids. You will use the earthworm as the model organism. Earthworms are also know as night crawlers, belong to the Oligochaeta, a clade of clitellate annelids that have become very common in the United states. Please compare the other living representatives to the living earthworm with regard to overall anatomy, feeding and locomotion.
You may be surprised to find that the common species used in biology classrooms is not a native. Lumbricus terrestris was probably brought to the US in the 18 th century. Their burrowing activities are thought to be beneficial by most gardeners. Unfortunately in forests, they redistribute nutrients, making them unavailable to other native detritus feeders and young plants, and in the process, are destroying the diversity of our forests. So although the burrowing activities of many polychaetes are beneficial and stabilize habitats such as mud flats, those of the night crawler for the most part are deleterious to our forest ecosystems.
4. External Anatomy and locomotion of the common night crawler
Obtain an earthworm, Lumbricus terrestris. Place it in a large dish to study its external anatomy and locomotion.
Orientation
Examine the external features of the worm. The anterior end is usually larger than the posterior and is round in cross section whereas the posterior tends to be flattened dorsoventrally

A dorsal view of the anterior end of Lumbricu s. The segments are numbered.
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). Anterior to the peristomium is a small lobe, the prostomium (pro = before) that, for embryological reasons , is not considered to be a segment.
The peristomium encircles the large, ventral mouth and is an almost complete ring around the body. 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 posterior most division of the body is the pygidium, which encircles the anus at the posterior tip of the worm. Like the prostomium, the pygidium is not considered to be a true segment.
Chaetae or Setae
The eight small chaetae or setae on each segment (except the first) are usually visible with adequate magnification (25X). Chaetae or setae are small chitinous bristles emerging from pores in the integument on the ventral half of the worm. The setae are arranged in four pairs, two 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.
A ventral view of Lumbricus.
The worm has been rolled slightly to reveal part of the right side.

The chaetae or setae 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 setae. Setae are amber or brown, slightly curved, and short.
Rinse your worm gently 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. Place it on a clean dish, and watch it move. Does it seem to have difficulty moving on the smooth glass surface. Place some stones and dirt in the dish and now watch it move. a. Explain why it was difficult for the earthworm with a muscle that essentially act to distort a hydroskeleton to move on the glass dish but not the dish that contains dirt and stones .
. b. When the earthworm moves in dirt
(or wet sand) do any segments move as a unit? c. Does the wave of contraction and relaxation move from front to back or back to front?
5. Lumbriculus variegates is a detritus feeder like the earthworm and a perfect example of severe miniaturization of form.
As always make sure you record your observations in your journal. a. Obtain a few mudworms. Place them is a small Petri or observation dish. Note how this Oligochaete moves. Also take one or two and put them on a wet paper towel.
Note the difference in “swimming” locomotion versus locomotion in an environment were they can contact a surface. b. You may be able at the highest power of your stereoscope to focus and watch them feed. Use the fish food provided.
Add a few tiny tiny crushed pieces of one flake, or one flake to the bowl. As always record your observation.
Notice how transparent at high power these worms are.
These worms are often used for experiments that look at the effects of drugs on cardiovascular function.

Place your worm in a plastic pipette and let it move to the tip.
Then place the tip of a capillary tube close to the tip of the pipette and gently squeeze. The worm should move into the capillary tube, which you can place then in a small observation dish. Observe the internal anatomy of the worm. Can you trace blood flow from dorsal to ventral? Time dorsal vessel contraction rate. c. Take a video or photographs of blood flow, showing the changes in vessel diameter. Any evidence for how these animals respire?
Then add a bit of alcohol (70 % ETHANOL ONLY-Alcohol is very toxic to invertebrates so start with 1ml or less in a dish 6 inches in diameter. ) to bowl or simply place your bowl on an ice block. d. What is the effect on heart rate? Describe the effect, noting whether you used ice or alcohol in your notebook.
If we had more time in this lab, we could test a host of other drugs and even determine a dose response curve. Do not place any worms exposed to alcohol or cold back in the colony dish.
Place them in a separate dish until we can determine if they will recover and not die and foul the colony dish..
6. Various small annelids . Examine several small species of annelids to document the effects of size on segmentation and the types of organs found in these species. These are Clitellates, and surprisingly most belong to Oligochaeta or the same clade as the much larger nightcrawler. .
Below are some photographs of some of the forms available.

Dero, back end, high power Yes that is the hind end, not the head, so what is that claw like structure?
Aeolosoma -often multiplies by fission, can get very long worms with several generations developing behind the parent generation.
So, are your cultures reproducing asexually?

Stylaria, Note large setae and prominent feeding structure at anterior.
So if that is what they use to feed, how do they feed?
Pick two forms, identify from the pictures which two forms you picked. a1 and a2 Photograph your two forms including their names in a corner of the photographs. b1 and b2. Compare them to Lubriculus in overall morphology and anatomy (number of segments, visibility of segments and clitellum, presence or absence of circulatory vessels, etc.). Make sure you take some of the green stuff in the culture with you, so you can watch them feed as well.
Below are some urls that may help you in identifying anatomy particular to a species, but do not simply download the photos or videos as your own. We will know!!!
www.microscopyu.com then go to galleries then pond life http://www.micrographia.com/specbiol/helmint/annelhom/olig01
00.htm#oligolink
The first few unidentified pics are probably aeolosoma. http://www.microbelibrary.org/index.php/library/resources/3190aeolosoma-hemprichi-ingestion-of-soil-particles-and-digestion-ofmicrobial-films

What general trends do you see? Record your observations in your journal. c. Produce a movie of locomotion in one of these species. How does the locomotion of your choice vary from that shown by
Lubriculus and the earthworm.
To examine these more closely, you may need to watch them under a depression slide or under slight compression on a regular slide. Only use low power, 10x or 20x so the slide can still hold enough water for these to move.
7. Hirudinea : Leeches are Clitellates, but not Oligochaetes. They belong to another group of Clitellates, known as the Hirudinea. We only have 4 specimens, so when you are done pass your specimens on.
Leeches are annelids modified for an ectoparasitic life style. The body has a terminal posterior sucker and a dorsal subterminal anus.
An anterior sucker is present in all but one species ( Acanthobdella peledina ).
As always make sure you record your observations in your journal.
Obtain a living leech and place it in some fresh water. Allow the leech to remain undisturbed for about 5 minutes in a well lighted room. Pass your hand over the container so that a shadow falls on the leech. a. Observe and record the leeches' actions. b. Allow the leech to remain undisturbed for at least 5 minutes.
Use a plastic ruler or tongue depressor to stir up the water surface. Observe and record the leech’s actions. What parts of the leech are receptive to water movement? Why would leeches have organs that are sensitive to water vibrations?

c. Watch it move and compare its movement with that of an earthworm. Do you feel the leech relies on its hydrostatic skeleton as much as an earthworm? Do you see types of movement, you would not expect in animal such as an earthworm without the “compact” body of a leech. d. Finally examine the leech’s external anatomy. Does it have more or less segments than an earthwork. Can it be divided into the same three areas? Does it have chaetae? e. Add a snail or small chunk of meat or earthworm to your dish.
Place it on the side of the dish furthest from the leech. How does the leech react? Watch your leech fed. If they are hungry enough they will feed on the beef even if they prefer earthworms or snails. You should see imprints of the jaws in the beef cube after feeding. If your specimen is cooperative, you will develop a respect for just how flexible a body a leech can possess.