The Western Flyer hunched into the great waves toward Cedros Island, the wind blew off the tops of the whitecaps, and the big guy wire, from bow to mast, took up its vibration like the low pipe on a tremendous organ. It sang its deep note into the wind.
A NOTE ON PREPARING SPECIMENS
Following are the methods of preparing, anesthetizing, and preserving specimens which we have followed. As previously remarked, some are by no means entirely satisfactory, but they are the best we know.
The chitons, which otherwise curl up tightly during preservation, can be removed directly from rock or collecting bucket and tied firmly to glass plates with string or strips of cloth. When they grip the plate, they are dropped into a container of ten-percent formalin; whereupon they are preserved relaxed.
Sipunculids (or peanut worms), to be preserved with the introvert extended, must first be anesthetized by sprinkling powdered menthol crystals on the sea water in a scrupulously clean glass dish. These worms are so delicate that metal or metal salts will cause them to draw in their heads immediately. The more refractory forms, after a few hours’ treatment with menthol, are killed by letting fresh water gradually replace the sea water. This must be done over a period of several hours. They are then preserved in formalin solution.
Alcyonaria, such as Renilla (the sea-pansy), Stylatula (the sea-pen), etc., are treated with Epsom salts for several hours until the expanded polyps are completely inert, then preserved in formalin if they are intended for display where color is important (although even thus the color is not successfully retained) or in alcohol if for study or identification.
Hydroids are relaxed with menthol for the small forms, with Epsom salts for the large gymnoblasts.
Anemones are very difficult to handle, and a perfect method for expanding them has not been developed. They are sensitive both chemically and physically. A specimen expanded and relaxed with Epsom salts will still draw in its tentacles if touched. Some results have been achieved by introducing pure oxygen directly into the stomach with a tiny hollow glass needle—a process which seems to intoxicate them. But it is at best a delicate and over-arduous operation. A fairly good method is gradually to introduce a saturated solution of Epsom salts into the pan where the animals are expanded, using a drip-string. This is followed later with novocain, or best of all, but usually unavailable, with cocaine, which is put into the water directly over the animal. Finally, formalin is introduced with a drip-string. Any shock, either chemical or physical, will cause immediate retraction of the tentacles. For permanent use a running seawater system which supplies O2 to the circulation of the animals being narcotized has proven most effective of all.
Crabs and shrimps of most types, and brittle-stars, are best killed by dropping them into fresh water as soon after they are captured as possible. After a brief struggle the crabs die without casting their legs, and the brittle-stars (with the exception of Ophioderma teres) will not curl up if this method is used. All these should subsequently be preserved in alcohol, since formalin will disintegrate the calcareous portions of the brittle-stars and soften the chitin of the crabs. For color notes only, specimens may be preserved in formalin.
Holothurians (or sea-cucumbers) are very delicate and have a tendency to eviscerate if kept in stale or over-warm water. They should be got as soon as possible into trays of cool, clean sea water, allowed to expand, then relaxed with Epsom salts used in considerable quantity. They should be preserved in alcohol. Most holothurians deteriorate badly in a few months if formalin is used on them, owing to the dissolution of the calcareous plates by which specialists determine them. Their neurotic tendency of deliberately casting out their viscera when they are sick or shocked or unhappy makes them difficult to handle. The cucumber itself, if times get better and a pleasant environment is restored, is able to grow a new set.
Starfish relax and puff out in a life-like manner when placed for several hours in fresh water. They can ordinarily be preserved in formalin, but museum specimens for determination should be placed in alcohol for a time and dried in the air. This method also applies to sea-urchins.
Sponges must not be placed in formalin even for a few moments. Entire colonies can be dried after a preliminary immersion in alcohol; small portions of the colonies so treated ought, however, to be preserved permanently in a vial of alcohol.
Flatworms such as the turbellarians are hard to collect, hard to handle, and hard to preserve. They are so delicate that the bodies are easily injured in picking them up. When they are crawling on a rock it is satisfactory to place a thin-bladed knife in their path; when the flatworm oozes onto the knife-blade he can be lifted into a container. Very small specimens may be lifted from the rock with a camel‘s-hair brush and transported to a glass plate. This is flooded with hot Bouin’s solution and immediately covered with another glass plate. If one could devise smooth plates permeable by Bouin’s solution it would be even better.
Several methods are applicable to the preservation of such pelagic invertebrates as jellyfish. Menthol crystals are a successful anesthetic for the contractile forms. However, the difficulties are likely to relate to the extreme softness of the more delicate forms. Hardening solutions of chromic acid and formalin, or osmic acid (which, however, is very expensive—five to six dollars a gram) in formalin, are most effective. Often there are mechanical difficulties involved in attempts to retain the original shapes of animals. The jellyfish Beroë tends to buckle up. We have had some success by inserting the closed end of a test-tube in the body cavity during the hardening process. It is really impossible to preserve jellyfish aboard a boat that is rolling even slightly. Formalin is usually a satisfactory preservative, and is often indicated exclusively.
It seems from these notes that all animals are difficult to preserve, and it is true that all of them require care which is not often enough given. The almost universal cry of specialists engaged in species determination is that specimens arrive in such bad condition that their work is made doubly difficult. Only extreme care can rectify this.
Some of the annelid worms are extremely difficult to handle. Anesthetizing methods are useful, but the delicacy of the animals and their constricting traits make special procedure necessary. Very long specimens may be wrapped around glass rods or test-tubes and suspended in formalin or alcohol, depending on the species. The chief difficulty with worms is to get the entire animal in the first place, especially the extreme anterior and posterior segments without which identification is difficult if not impossible. Some of the worms sting very badly and should not be handled at all.
For the preservation of fishes, formalin in general gives better results than alcohol. Small specimens are hardened by putting them directly into trays of formalin solution; while larger fishes must have the body cavities injected with a twenty-to twenty-five-percent solution of formalin to which a little glycerin has been added.
Formalin should be used as follows: U.S.P. formaldehyde solution of thirty-eight to forty percent is a gas dissolved in water, and this percentage represents saturation. One part of this to sixteen parts of water for small specimens is successful when the amount of the solution is many times the bulk of the animal. Large specimens are hardened in trays of ten- to fifteen-percent solution. One must check by experiment on the particular animal involved, feeling it after a few hours to note the consistency of the tissue. Formaldehyde is very irritating to nose, lungs, and skin. Rubber gloves should always be used and the solution worked with in large rooms or in the open air. The tolerance of a person working with formaldehyde sometimes decreases with time, so that he is sickened at the odor or even breaks out with allergy eruptions on contact or association.
With alcohol the ultimate preservation is usually in seventy-percent strength. For crabs and so forth in the tropics or in very hot weather, glycerin should be added. This keeps the animals flexible and less brittle—less likely to break up—and also prevents poor preservation due to bubbles forming in the solution. For ideal preservation the spec
imen should be brought immediately after killing into a twenty-five-percent solution, then to fifty percent and finally to seventy percent. Large specimens, where the amount of liquid is small in proportion to the amount of tissue involved, may require ninety-percent solution for a few days of hardening before they are placed in a new seventy-percent solution.
Labeling is easy, simple, and necessary. Yet the failure to label clearly and immediately has led to many ridiculous situations. One expedition, which need not be named, labeled Atlantic animals as coming from the Panamic regions. And another completely lost track of its collection, to the disgust of the specialists who later tried to determine the species. Labels are best made on slips of good drawing paper and printed with a drawing pen in India ink. Each label should include the date, the exact place, the depth, and a number added which will agree with the number in the collecting notes. In the collecting notes, under this number, should occur any remarks covering ecological factors or observed action of the living animal which would be impossible to put on the label. The label should be placed inside the jar with the animal, and it should be done immediately, before a new lot of specimens comes in. There has not, to our knowledge, been any single expedition or extended trip which failed to turn out some unlabeled, or mislabeled material, so that the records are full of obviously incorrect reports. Some Panamic animals have been reported from Puget Sound, and our common California shore crab, Pachygrapsus, was originally described as from the Sandwich Islands. Immediate labeling, on the same day as the collection, is the only way to reduce these errors to a minimum. This cannot be over-emphasized.
Fig. 1. Phyllonotus bicolor (Valenciennes ) 1852 § S-358 Pink Murex
Fig. 1. Murex rectirostris Sowerby. Gulf spiny Murex § S-355
Fig. 2. Cerithium maculosum Kierner § S-374
The Spotted Cerithium
Fig. 1. Thais tuberculata Gray. Knobby Thais § S-367
Fig. 2. Phyllonotus princeps (Broderip) § S-361
Regal Murex
Fig. 1. Protothaca grata (Sowerby) § S-252
Varicolored Edible Clam
Fig. 2. Cypraea annettae Dall 1909. Brown Cowry § S-370
Fig. 1. Terebra variegata Gray 1834 § S-326
Variegated Augur-shell
Fig. 2. Nassarius ioaedes and luteostoma § S-340 and § S-341
Fig. 1. Callopoma fluctuosum (Wood) 1828 § S-404
Common Gulf Turbine-shell
Fig. 2. Oliva venulata Lamarck § S-331
Gulf Olive-shell
Fig. 1. Gulf Olive-shell. Albino color variety § S-331
Fig. 2. Chiton virgulatus Sowerby 1840 § U-17
Common Gulf Chiton
Fig. 1. Strombus gracilior Sowerby 1825 § S-372
Smooth Conch
Fig. 2. Melongena patula § S-338
Broderip and Sowerby
Fig. 1. Ophiocoma alexandri Lyman 1860 § K-215
½ natural size
The drawings on plates 9-24 inclusive are by Alberté Spratt
Fig. 1. Nidorellia armata (Gray) 1840 § K-126
natural size, 5½“-6” oral surface
Fig. 2. Holothuria impatiens (Forskål) 1775 § L-21
½ natural size
Fig. 1. Oreaster occidentalis Verrill 1867 § K-125
natural size, 6”
Fig. 2. Nidorellia armata (Gray) 1840 § K- 126
natural size, 5½“-6” aboral surface
Fig. 1. Petrochirus califomiensis Bouvier 1895 § Q-16
¾ natural size
Fig. 2. Tedania ignis (Duchassaing and Michelotti) 1864 § A-23
½ natural size
Fig. 1. Ophiocoma acthiops Lütken 1859 § K-214
aboral surface. ½ natural size
Fig. 2. Dentalium semipolitum Broderip and Sowerby § S-103 and 104 and D. splendidum Sowerby 1832 1¼ natural size
Fig. 1. Ocypode occidentalis Stimpson 1860 § R-75
½ natural size
Fig. 2. Callinectes bellicosus Stimpson 1859 § R-42
½ natural size
Fig. 1. Phataria unifascialis (Gray) 1840 § K-120
½ natural size
Fig. 2. Ophiactis sp.
1½ natural size
§ K-218 (219?)
Fig. 3. H. lubrica Selenka 1867
Sulphur cucumber § L-25
½ natural size
Fig. 4. Luidia pbragma Clark 1910 §K-128
natural size, 4½“-5”
Fig. 1. Clibananus panamensis § Q-14
Stimpson (1859) 1862 natural size
Fig. 2. Phycosoma antillarum (Grube and Örsted) 1859 1-10 natural size
Fig. 3. Othilia tenuispina (Verrill) 1871 as Echinaster § K-124
Fig. 1. Clypeamr rotundus (A. Agassiz) 1863 § K-322
½ natural size
Fig. 2. Echinometra vanbrunti A. Agassiz 1863 § K-314
½ natural size
Fig. 1. Eurythoë complanata (Pallas) 1766 § J-35
⅔ natural size
Fig. 2. Geograpsus lividus § R-68
(Milne-Edwards) 1837 ½ natural size
Fig. 3. Portunus minimus § R-40
Rathbun 1898 natural size
Fig. 4. Eriphia squamata § R-58
Stimpson 1859 1½ natural size
Fig. 5. Daira americana § R-45
Stimpson 1860 natural size
Fig. 6. Trapezia cymodoce ferruginea
Latreille 1825 1½ natural size § R-60
Fig. 1. H. paraprinceps § L-26
Deichmann 1937 ½ natural size
Fig. 2. Meoma grandis Gray 1852 § K-324
½ natural size
Fig. 3. Astropyga pulvinaca (Lamarck) 1816 § K-318
Fig. 1. Pharia pyramidata (Gray) 1840 § K-119
Fig. 2. Chloeia viridis § J-34
(Schmarda) 1861 ⅔ natural size
Fig. 3. Notopygos ornata
Grube 1856 § J-36
natural size
Fig. 1. Evibacus princeps § P-105
Smith 1869 ¾ natural size
Fig. 2. Callopoma fluctuosum § S-404
(Wood) 1828 natural size
Fig. 3. Pocillopora capitata Veirill 1864 § E-7
¾ natural size
Fig. 4. Hypoconcha panamensis
Smith in Verrill 1869 § R-25
¾ natural size
Fig. 5. Pontonia pinnae Lockington (1878) 1879 natural size § P-25
Fig. 6. Percnon gibbesi § R-74
(Milne-Edwards) 1853 natural size
Fig. 1. Eucidaris thouarsii § K-313
(Valenciennes in L. Agassiz and Desor) 1846
½ natural size
Fig. 2. Petrolisthes edwardsii (Saussure) 1853 § Q-27
½ natural size
Fig. 3. Cliona celata Grant 1826 § A-13
⅓ natural size
Fig. 1. Leucetta losangelensis A-18
(de Laubenfels) 1930 ½ natural size
Fig. 2. Arbacia incisa § K-315
A. Agassiz 1863 ½ natural size
Fig. 3. Mithrodia bradleyi Verrill 1867 ½ natural size § K-122
Fig. 1. Heliaster kubiniji Xantus 1860 § K-116
Fig. 2. Thoë sulcata § R-32
Stimpson 1860
PHOTO BY HAMJUOD L. SWIFT
Fig. 3. Mellita longifissa Afichelin 1858 1½ natural size § K-323
Fig. 1. Cerithidea mazatlanica Carpenter 1857 § S-373
Mazatlan Horn-shell
PHOTO BY HARLAND L. SWIFT
Fig. 2. Octopus sp. upper surface
Diameter of body 49.1 mm.
Fig. 3. Octopus sp. under surface
§ U-116
PHOTOS BY WILLIAM G. VESTAL
Fig. 1. Pilumnus gonzalensis § R-52
Rathbun 1893
Fig. 2. Pilumnus townsendi § R-54
Rathbun 1923
PHOTO BY HARLAND L. SWIFT
Fig. 3. Siphonaria pica Sowerby. Starry Pulmonate § S-325
Limpet, under surface
Sea of Cortez Page 27