To find the animals the amateur must first become well acquainted with their various habitats: rocky shores, sandy beaches and marine eelgrass meadows. He must also know the proper methods of preservation. The techniques employed for preserving lifelike specimens are not necessarily easy to master. Some organisms, particularly the more fragile invertebrates, are easily damaged during capture and tend to assume grossly distorted forms when treated with harsh chemicals. Others, such as brittle stars, literally break into pieces when they are excited. According to Jack J. Rudloe, a former amateur collector who now operates the Gulf Specimen Company in Panacea, Fla., the collection and preservation of marine specimens is largely an art, based on an admixture of zoology, biology and biochemistry.

"The sand and surf," Rudloe writes "tend to keep the secrets of their marine animals hidden from the casual vacationer at the seashore, but actually all one requires to find the teeming life at the edge of the sea is curiosity and observation. Every little ridge, hump, hole and trail in the sand points the way to a beach creature. The techniques of collecting tend to vary according to the nature of the habitat-rock, sand or grass.

"Rocky shores are home to animals that have the ability to cling tenaciously to their perches to keep from being dislodged by the battering surf. Rather than loosen their hold many rock-dwelling forms will allow themselves to be ripped to pieces. As a result they present certain problems to collectors. Typical examples include removing a polyclad flatworm or a nemertean worm from a rock without fragmenting the creature, and picking up a sea cucumber without having the specimen nervously expel its viscera. The acquisition of techniques for coping with such problems makes all the difference between catching and losing the animals.

"To dislodge rock-dwelling animals such as limpets, abalones and chitons without damaging them one must pry them off tediously with a flat-bladed knife. Barnacles can pose a particularly difficult problem: such forms as Balanus balanoides, Chthamalus and Tetraclita have only a membranous base, and one cannot remove them from rocks without breaking the shell or tearing the gonads. Even an experienced collector will crack more barnacles than he cares to. The best approach is to chip off a piece of rock with barnacles growing on it, using a geologist's hammer; alternatively one might seek the same species Of barnacle on more manageable surfaces such as wood or shell.

"Often the type of rocky surface determines the method of collecting the particular animal. Sandstone, common in the Pacific, permits digging, and one can chip out an animal without too much trouble. On hard rock the nature of the surface regulates the collecting approach. For example, one can occasionally collect polyclad flatworms found on smooth, hard rock by letting them ooze onto the blade of a knife. The knife is useless, however, on rough surfaces because the flatworm hides in crevices and generally breaks, tatters and finally disintegrates if the collector persists.

"A technique I have found useful is to mold a plastic bag over the surface of the rock and gently chase the flatworm into the opening by splashing the creature lightly with water. The plastic bag method has worked well with other sensitive invertebrates. A squirt gun filled with weak isopropyl alcohol will drive out many of the shrimps, amphipods, polychaete worms and brittle stars that dwell in deep crevices.

"On the upper limits of the slippery rocks and cliffs are hundreds of scurrying isopods, or multilegged crustaceans, which can cause the inexperienced collector much difficulty. The novice may try to climb the dangerous rocks, slapping at the creatures. Even if he catches any, the delicate specimens may end up squashed. I have found insecticide spray effective in stopping these animals in their tracks. After it is applied they can be dropped directly into a vial of alcohol.

"When the tide recedes, undisturbed sea anemones hang flaccid. If a collector quickly jerks them, they generally come right off the rock. Speed is necessary or they will whip in their tentacles and contract immediately into a tough, fast-sticking bulb. Some species even burst their body walls. If the rocky surface cannot be broken away from the anemone's basal disk, pounding the rock next to the specimen with a geologist's hammer will do the trick: the vibration causes the anemone to fall off.

"Much collecting in rocky areas can be done by hand. Among the creatures thus obtainable are nudibranch mollusks, polychaetes, sponges, tunicates, hydroids, hermit crabs and snails. Sea anemones and sipunculid worms, however, prefer deep crevices and crannies and are often impossible to pull out. By turning over rocks with a crowbar (wearing heavy gloves to protect the hands from cutting shells and the calcareous spicules of sponges) one can obtain assorted varieties of sea cucumbers, echiuroid worms, mud shrimps and even small eels. Octopuses live in burrows in temperate zones and can be spotted by the heaps of crab shells they have discarded. Pinching crabs and cutting mantis shrimps should be cautiously handled or captured with a small dip net.

"The beautiful sabellid worms known as feather dusters pull in their tentacles at the slightest disturbance and withdraw into their tubes. Gently pry the tube from the underside of the rock. Always remember to return the rock to its natural position or entire animal and plant communities will die needlessly from exposure.

"Although collecting marine animals on rock piles is often easier than combing miles of sandy, muddy beach, the sandy habitats give rise to a diverse and complex community of animals that presents a challenge to the amateur collector. Almost every little creature is adapted to burrowing. Some, such as the translucent lancelet amphioxus, can move through hard-packed sand almost as quickly as a fish swims through water. Others that move slowly over sand, such as the frilled sea hare, Bursatella leachi, have protective coloration. There are many bivalves and worms that avoid sunlight and stay buried.

"To find the inhabitants of sandy areas one must do some sleuthing. The clues are peculiar slits and trails in the sand. These will vary, of course, with the animal and with the type of habitat. At low tide you can find numerous polychaetes, moon snails, little portunid crabs and an occasional sea cucumber.

One must know not only how to recognize an animal but also where it hides and when it is most likely to show itself.

"On most outer beaches where the rolling waves wash up, large numbers of the small, multicolored coquinas (Donox) burrow rapidly into the drenched sand to escape their many predators. Every receding wave exposes the little clams and they busily begin burrowing again. The amateur collector soon learns to recognize their presence by the thousands of tiny pepper-shot holes. By screening or handpicking he can easily fill a bucket. The same surf tumbles mole crabs (Emerita) about in the water for a second and they burrow down again swiftly. With one sweep of the net you can build up a good series of specimens.

"Air and water temperatures influence the mobility of marine animals. It is often much easier to collect the rapidly burrowing forms at low tide in freezing winter weather than in the summer. The burrowing sea anemone (Bunodactis stelloides) rapidly draws in its tentacles and sucks down into the sand when lightly touched, but in winter it contracts only slightly while being spaded up. The cerianthid anemone known locally as 'sloppy guts' spins its own tough, leathery tube with mucus, sand and mud. The animal rides up and down this long tube as if it were an elevator. In warm weather, even at the slightest disturbance, the anemone will rocket down to the bottom. The collector must dig slowly. One impatient yank may leave him with an empty tube. In winter when the air temperature is below freezing, however, it is possible to sever the tube and block the anemone's descent with a shovel before the animal can respond to stimuli.

"The eelgrass in enclosed sandy bays and estuaries shelters an abundance and diversity of animal communities all year. At spring tides that are windblown, so that the ribbed sand is exposed far beyond the normal littoral, the flats are strewn with castings of polychaete worms, balanoglossid worms, callianassid mud shrimps and other animals adapted for burrowing. The huge, reddish-black lugworm (Arenicola marina) tunnels a foot or two below the surface, making its presence known by little heaps of digested mud. It takes two collectors digging toward each other from both ends of the long burrow to capture a single specimen.

"Often in the sandy shore habitats there are mysterious creatures, probably crustaceans of some kind, whose burrows are so deep that the most energetic, rapid shoveling will not expose them. From the Sakalava natives of Madagascar I learned one effective trick for removing the deep-burrowing sipunculid worms. The natives insert a long flexible wire all the way down the hole until they feel the animal squirming. The holes are deceptive: they go straight down for about a foot and then level out horizontally before ending in another opening. After locating the worm with a wire the Sakalava, who use sipunculids for fish bait, speedily dig down to the exact location of the worm and yank it out.

"Another clue to hidden creatures is the sudden spurt of water one may see or hear as one walks along the exposed beds of sandy or muddy shores. Such a spurt may lead you to clams, mussels, sea cucumbers or sea squirts (Styela plicata). Often the animals are so well camouflaged that one might overlook them if it were not for the spurts.

"In eelgrass I have been challenged by the task of finding the small, elusive brachiopods that are often known as lamp shells. When I first tried, I would occasionally bring one up in a shovelful of sand while I was digging over a large area. In time I came to recognize their knifelike slits in the sand among the eelgrass roots, and I learned that they live close to the surface. Since then I have been able to collect hundreds of these animals in just a few square yards by turning them out with a table fork.

"Along the mud flats of protected lagoons and bay shores of the southeastern U.S. one can collect hundreds of the common sea cucumbers (Thyorlella gemmata and Thyone briareus) when they expand their floriated crowns up out of the mud at night to feed. Quickly clamp your fingers behind the expanded tentacles of the cucumber, strangling it so that it cannot draw its crown back into its body. Then tie a rubber band tightly behind the tentacles to keep them expanded and immerse the cucumber in 95 percent ethyl alcohol. Do not pull or tug too hard because the animal has the habit of bursting its body walls and ejecting its viscera.

"By sweeping a fine-meshed net along the bottom of the waving eelgrass at high tide one can find an abundance of shrimps, hermit crabs, snails, gobies, pipefish, flatworms and tunicates in an astonishing variety. Surprisingly, dipnetting in the eelgrass meadows is one of the least known but most productive methods of collecting.

"Collecting is absorbing work and can be exhausting. It is easy for a collector to work beyond his normal strength and endurance. He should guard against doing so, because collecting is only part of a day's work. It is also necessary to preserve specimens promptly in order to stabilize the tissues against drastic chemical change. Much material obtained at considerable effort has been ruined because the tired collector postponed caring for it until the next day. Some specimens can be carried alive for a time; a good way to do so is to put the animal in a plastic bag half-filled with seawater, seal the bag and then carry it in a bucket containing some seawater for cushioning. Other specimens, such as sponges, must be preserved immediately or they will begin to undergo a process of deterioration that results in physical distortion and loss of the flagellate chambers. Although no animal will look the same after it has been preserved, it will look similar to its living self if the preserving is done with care. The collector has a responsibility: he knows how the animal looked when it was alive and must duplicate it as best he can.

"Sponges, starfish, sea urchins, crabs and shrimps may be placed directly in preservative Brittle stars, sea spiders and certain crabs, however, often throw off their legs in their violent death struggle, so they are best killed by simply adding weak alcohol or fresh water to their seawater until they succumb. The feathery legs of barnacles can be extended in the same way. The trick is not to become impatient and pour in the killing agent too quickly.

"The highly sensitive sea anemones, hydroids, nudibranchs, tectibranchs, echiuroids, sipunculids and tunicates present real difficulty. Years ago, when I first began collecting, I had repeated failures in preserving such contractile invertebrates. I took my problem to Charles E. Cutress, a specialist in sea anemones at the U.S. National Museum in Washington, and he suggested some simple and inexpensive methods that are very effective. They involve anesthetizing the specimens before killing them.

"Three of the better anesthetics are Chloretone, menthol crystals and Epsom salts (magnesium sulfate). The first two are scarcely soluble in water and should be used in small amounts. A few crystals added to a dish of specimens will suffice. Magnesium sulfate is extremely soluble in water and is used in a saturated solution. To avoid disturbing the animals the Epsom salts should be tied in a piece of porous cloth, and only a corner of the bag should be allowed to touch the water. Diffusion will take place so slowly that the animals will not be agitated while going to sleep.

"Before adding an anesthetic to a dish of specimens make sure they are naturally expanded. Sea anemones, siphonophores, hydroids and gorgonians must have their tentacles or polyps distended, the gills of nudibranchs must be out and the introverts of sipunculids and echiuroids must be fully extended. Orifices of sea squirts should be open as they are when naturally siphoning the water; sometimes a glass rod can be used to support the openings.

"The time it takes to render specimens insensitive will vary widely. No two specimens will become insensitive at the same time. It is best to treat them individually. Never mix species of coelenterates because cross-contamination of their nematocysts, or stinging organs, makes the process of anesthetization twice as difficult. Some contractile creatures such as small anemones and hydroids may be rendered insensitive in an hour, certain species of polychaetes and sipunculids may take six hours and nudibranchs sometimes require 15 hours before they are ready for preservation. When the animal does not react to probing, it is ready for preservation. The animals should not be allowed to die in the narcotizing fluid; instead they should be preserved shortly after immobilization. When you have done a considerable amount of preserving, you will develop a sense of the right moment for fixing the specimen.

"Remove most of the- water from the dish, leaving only enough to cover the organism. Then flood in full-strength formalin (37.9 percent) and the specimens will usually die in an expanded condition. There are always a few specimens, however, that gather up enough energy to contract at the last second. For this reason I prefer the old method of formalin poisoning I learned at the Marine Biological Laboratory in Woods Hole, Mass. The contractile specimens are first partially narcotized in a large volume of seawater and then a few drops of dilute formalin (three drops of 1 percent formalin per 100 cubic centimeters) are added at 15-minute intervals. In this process death results much more quickly and there is less chance of disintegration.

"With certain nemerteans and polychaetes, notably the sabellids, specimens are best narcotized by slowly adding alcohol at 15-minute intervals until they become insensitized. Otherwise nemerteans are apt to vomit their long proboscis and sabellids to throw off their feathery tentacles. Formalin poisoning will usually kill the animals in an expanded condition after the alcohol has immobilized them.

"Enthusiasts who go in for shell collecting face no problems. Simply boil the animal until the flesh is loosened, then scoop out the meat and allow the shell to dry. Those interested in preserving the whole animal will need the same patience as the collector of soft forms. I use a simple and inexpensive technique to get the best results. I boil a pan of seawater for a few minutes to drive off all the oxygen. When the water has cooled to room temperature, I place the mollusks in a flat, shallow pan for immersion in the cooled water. The specimens soon expand their meaty bodies far out of their shells in an effort to absorb the oxygen that is not there. When the soft parts have been extended to their natural feeding and moving positions, I place the pan in the freezer, along with a few jars of 70 percent ethyl alcohol. By the time the mollusks are frozen solid the alcohol is chilled to 25 degrees Fahrenheit or less. I remove the mollusks and place them in the prechilled alcohol, where they're remain frozen solid, but the alcohol penetrates their tissues thoroughly and they die perfectly expanded. I learned from experience that if frozen snails are placed in alcohol at room temperature, they quickly thaw out and manage to withdraw completely into their shells before they die. Many corals and bryozoans can be killed in a similar manner. First they are narcotized until the polyps are expanded; then they are frozen and strong formalin is poured over the ice. As the ice thaws, the formalin mixes slowly with the water and penetrates and kills the specimens.

"There are more little tricks to learn. Polyclad flatworms generally explode no matter what preservative is used. I found that the tropical Indian Ocean forms could be fixed nicely if they were first relaxed with menthol and then dropped into hot nitric acid. Death is so instantaneous and the tissues are hardened so quickly that there is no time for the animals to contract. Nitric acid is also a good fixative for ctenophores, the comb jellyfish that are impossible to preserve otherwise. In both cases the specimens are washed in running seawater until all the fixative is washed out and are then preserved in 5 percent formalin.

"Denatured ethyl alcohol is a good general preservative for most of the invertebrates. Sponges, mollusks, crustaceans, echinoderms and tunicates can all be preserved directly in alcohol, but always make sure there is plenty of preserving fiuid in the body cavity to prevent internal maceration. Sea anemones and jellyfish should never be put in alcohol because the solution will dehydrate and distort them. Therefore a 10 percent formalin solution (nine parts of seawater to one part of concentrated formalin) is recommended. Formalin, on the other hand, should never be used in preserving any starfish, sea urchin, sea cucumber, sponge or horny coral because the acid breaks down the calcareous spicules, rendering the specimen useless for identification. If, however, the specimens are preserved in neutralized formalin (one pound of hexamethylenetetramine added to one gallon of commercial formaldehyde), they will keep better and even retain some of their original color. Unfortunately no procedure has been developed for preserving intact the brilliant colors of marine fishes and invertebrates.

"After preserving a specimen be sure to include on an accompanying label data about the color, size before preservation and any other features that may seem diagnostic or liable to be lost in preserving. The label should also state where the specimen was collected, the date, the depth of the water and the collector's name. An unlabeled specimen floating in harsh preservative cannot supply this information.

"Preparing and preserving marine animals requires endless time and patience. A specimen can be well expanded and thoroughly preserved but still not be a good specimen. Specimens must be good before preservation. A squid with all its tentacles and no torn parts, a sea urchin with no broken spines or a starfish with all its arms is classed as a good specimen. If the animal does not look right before it is preserved, it certainly will not improve with age. The collector has approached perfection if his specimen is well extended and relaxed and retains at least some of its natural color."

Bibliography

THE EDGE OF THE SEA. Rachel Carson. Houghton Mifflin Company, 1955.

FIELD BOOK OF SEASHORE LIFE. Roy Waldo Miner. G. P. Putnam's Sons, 1950.

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