%PDF-1.4 % 1 0 obj << /Type /Catalog /Pages 2 0 R /AcroForm 34 0 R /Metadata 52 0 R >> endobj 2 0 obj << /Type /Pages /Kids [ 3 0 R 10 0 R 17 0 R 24 0 R ] /Count 4 >> endobj 3 0 obj << /Type /Page /Parent 2 0 R /Resources 4 0 R /Contents 5 0 R /Rotate 0 /eCpyResolution 300 /MediaBox [ 0 0 612 1008 ] /CropBox [ 81.36 342 540 1008 ] /Annots 33 0 R >> endobj 4 0 obj << /XObject << /Im6 6 0 R >> /Font << /F7 7 0 R /F8 8 0 R /F9 9 0 R >> /ProcSet [ /PDF /Text /ImageB /ImageC ] >> endobj 5 0 obj << /Length 4626 >> stream q q 612.00 0.00 0.00 1008.00 0.00 0.00 cm 0.00 0.00 0.00 rg /Im6 Do Q Q q BT 3 Tr /F7 12.65 Tf 276.00 904.80 TD (NEMERTEA) Tj /F8 6.93 Tf -9.12 -20.64 TD (C erebratulus lacteus) Tj /F8 8.70 Tf -135.12 -19.44 TD (LIVING MATERIAL:) Tj /F7 11.07 Tf 15.12 -14.88 TD (Adults are found near or below the low-water mark, burrowing in mud, sandy) Tj /F7 11.33 Tf -14.88 -11.04 TD (mud, or sand in sheltered bays, harbors and estuaries. If not available at Woods) Tj /F7 11.30 Tf 0.24 -11.04 TD (Hole they can be shipped in from Maine or from north of Cape Ann, Mass. The) Tj /F9 9.88 Tf 0.00 -11.04 TD (sexes are separate. Sexually ripe males are bright red, ripe females dull, brownish) Tj /F9 9.81 Tf 0.24 -11.04 TD (red; in spent animals these colors fade \(Coe, i 89 5\) .) Tj /F8 8.75 Tf 0.24 -18.96 TD (BREEDING SEASON:) Tj /F7 11.78 Tf 15.60 -14.88 TD (May and June in Long Island Sound, July in the Woods Hole region, and) Tj /F9 10.02 Tf -15.84 -13.68 TD (July and August in Massachusetts Bay and on the Maine coast \(Coe, i 937\) .) Tj /F7 8.96 Tf 0.72 -16.56 TD (PROCURING AND HANDLING MATERIAL:) Tj /F8 7.86 Tf 14.88 -14.88 TD (A. Care of Adults: The sexes are best segregated. The female Cerebratulus) Tj /F7 11.18 Tf -14.88 -11.28 TD (may be kept for three weeks or more in the laboratory, but the eggs are less suit-) Tj /F7 11.50 Tf 0.24 -11.28 TD (able for experimental purposes after the first week. The worms should be kept) Tj /F7 11.79 Tf 0.24 -10.80 TD (in tall battery jars, over the tops of which gauze covers are securely tied; the) Tj /F7 11.22 Tf 0.00 -11.28 TD (worms are active and will escape down the drain unless this precaution is taken.) Tj /F7 11.66 Tf 0.24 -11.04 TD (A sea water hose should extend through the gauze to the bottom of the jar, to) Tj /F7 12.23 Tf 0.00 -10.80 TD (insure a continuous supply of fresh sea water. A low temperature \(around) Tj /F7 12.34 Tf 0.72 -11.04 TD (10 C.\) is optimal, but it is diffcult to maintain with running sea water at) Tj /F9 9.98 Tf -.72 -11.28 TD (Woods Hole.) Tj /F7 11.34 Tf 15.12 -10.56 TD (B. Procuring Gametes: A continuous supply of gametes may be obtained by) Tj /F8 7.39 Tf -14.64 -11.28 TD (removing successive portions from the posterior end of a ripe individuaL. A large) Tj /F7 10.98 Tf -.24 -11.28 TD (female, for example, may produce upwards of fifty million eggs.) Tj /F7 11.21 Tf 15.12 -10.80 TD (Female gametes: Females kept in the manner described above wil shed some) Tj /F7 11.26 Tf -14.40 -11.52 TD (eggs spontaneously, but to obtain large numbers a posterior piece, an inch long,) Tj /F8 5.00 Tf 0.24 -11.04 TD (should be cut off and placed on a five-inch square of cheesecloth in a finger ) Tj /F7 9.96 Tf 339.12 4.08 TD (bowl) Tj /F7 11.34 Tf -339.36 -15.36 TD (of cool sea water. With scissors or scalpel make a slit on either side of the mid-) Tj /F8 7.68 Tf 0.24 -11.04 TD (dorsal line. The muscular contractions of the fragment wil soon force the ripe) Tj /F8 7.39 Tf 0.00 -11.28 TD (ova into the water. The cheesecloth retains the slime and body section and allows) Tj /F7 11.90 Tf 0.24 -10.80 TD (the eggs to filter through. When they have settled, decant the water and add) Tj /F7 10.91 Tf 0.00 -11.04 TD (fresh, filtered sea water.) Tj /F7 11.47 Tf 15.12 -10.80 TD (Male gametes: To obtain sperm, a half-inch posterior fragment of a male is) Tj /F9 10.03 Tf -14.88 -11.04 TD (placed in a dish of clean sea water and a puncture made through the dorsal body) Tj /F7 11.13 Tf 0.24 -11.28 TD (walL. A drop of sperm suspension, drawn up into a fine-mouthed pipette, should) Tj /F7 11.01 Tf 0.00 -11.04 TD (be diluted in 40 cc. of sea water.) Tj /F7 11.07 Tf 15.84 -10.80 TD (C. Preparation of Cultures: When first shed, the egg contains a germinal vesi-) Tj /F7 10.89 Tf -15.36 -11.28 TD (cle, and fertilization at this time results in polyspermy and abnormal development.) Tj /F8 7.78 Tf 0.24 -11.04 TD (Allow the eggs to stand until microscopic examination shows that the germinal) Tj /F7 11.97 Tf 0.00 -11.28 TD (vesicles have ruptured and the eggs have reached the metaphase of the first) Tj /F7 11.21 Tf 0.24 -11.04 TD (maturation division \(a matter of 10 to 30 minutes, depending on the temperature) Tj /F9 10.81 Tf 0.24 -11.28 TD (and the ripeness of the eggs\). Now add one drop of freshly prepared, dilute) Tj /F7 8.16 Tf 175.68 -12.48 TD (44) Tj /F7 5.00 Tf 62.64 -103.92 TD ('-'.~""...) Tj /F9 5.00 Tf -213.12 -91.68 TD (";"::~_~::~~~.;i':'~I::~'" --.) 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