THE
BIOLOGICAL BULLETIN
PUBLISHED BY
THE MARINE BIOLOGICAL LABORATORY
Editorial Board
GARY N. CALKINS, Columbia University
E. G. CONKLIN, Princeton University FRANK R. LlLLIE, University of Chicago
E. N. HARVEY, Princeton University CARL R. MOORE, University of Chicago
SELIG HECHT, Columbia University GEORGE T. MOORE, Missouri Botanical Garden
LEIGH HOADLEY, Harvard University T. H. MORGAN, California Institute of Technology
M. H. JACOBS, University of Pennsylvania G. H. PARKER, Harvard University
H. S. JENNINGS, Johns Hopkins University W. M. WHEELER, Harvard University
E. E. JUST, Howard University EDMUND B. WILSON, Columbia University
ALFRED C. REDFIELD, Harvard University Managing Editor
VOLUME LXI
AUGUST TO DECEMBER, 1931
Printed and Issued by
LANCASTER PRESS, inc.
PRINCE 8C, LEMON STS. LANCASTER, PA.
11
THE BIOLOGICAL BULLETIN is issued six times a year. Single numbers, 51.75. Subscription per volume (3 numbers), $4.50.
Subscriptions and other matter should be addressed to the Biological Bulletin, Prince and Lemon Streets, Lancaster, Pa. Agent for Great Britain: \Vheldon & Wesley, Limited, 2, 3 and 4 Arthur Street, New Oxford Street, London, \V.C. 2.
Communications relative to manuscripts should be sent to the Managing Editor, Marine Biological Laboratory, Woods Hole, Mass., between May 1 and November 1 and to the Institute of Biology, I >ivinity Avenue, Cambridge, Mass., during the remainder of the year.
Entered October 10, 1902, at Lancaster, Pa., as second-class matter under Act of Congress of July 16, 1894.
LANCASTER PRESS, INC. LANCASTER, PA.
CONTENTS
No. 1. AUGUST, 1931
PAGE
Thirty-third Report of the Marine Biological Laboratory 1
TYLER, ALBERT
The Relation between Cleavage and Total Activation in Arti- ficially Activated Eggs of Urechis 45
SMITH, GEORGE MILTON
The Occurrence of Melanophores in certain Experimental Wounds of the Goldfish (Carassius auratus) 73
DICKMAN, ALBERT
Studies on the Intestinal Flora of Termites with reference to their Ability to Digest Cellulose 85
LUTZ, BRENTON R.
The Innervation of the Stomach and Rectum and the Action
of Adrenaline in Elasmobranch Fishes 93
JOHNSON, GEORGE E., AND NELSON J. WADE
Laboratory Reproduction Studies on the Ground Squirrel, Citellus tridecemlineatus pallidus, Allen 101
BURKENROAD, M. D.
A New Pentamerous Hydromedusa from the Tortugas 115
ONORATO, A. R., AND H. W. STUNKARD
The Effect of certain Environmental Factors on the Develop- ment and Hatching of the Eggs of Blood Flukes 120
No. 2. OCTOBER, 1931
PATTERSON, J. T.
Continuous versus Interrupted Irradiation and the Rate of Mutation in Drosophila 133
TORVIK, M. M.
Genetic Evidence for Diploidism of Biparental Males in Ha- brobracon 139
WTERNER, ORILLA STOTLEK.
The Chromosomes of the Domestic Turkey 157
ALEXANDER, GORDON
The Significance of Hydrogen Ion Concentration in the Biology
of Euglena gracilis Klebs 165
iii
38883
iv CONTEXTS
REDFIELD, A. C., AND M. FI.ORKIX
The Respiratory Function of the Blood of Urechis caupo . ... 185
SCOTT, \Y. J.
Oxygen and Carbon Dioxide Transport by the Blood of the Urodele, Amphiuma tridactyla . . . .... 211
MAST, S. O.
Movement and Response in Ditrlugia with special reference
to the Nature of Cytoplasmic Contraction 223
STUNKARD, H. \Y.
The Effect of Dilution of Sea Water on the Activity and Lon- gevity of Certain Marine Cercariae 242
No. 3. DKCKMUKR, 1931
HARVEY, E. NEWTON
The Tension at the Surface of Marine Eggs, especially those
of the Sea Urchin, Arbacia 273
TAYLOR, G. WKLLFORD, AND E. NEWTON HAKVKY
The Theory of Mitogenetic Radiation . . 280
YYlHTAKKk, D. M.
Some Observations on the Eggs of Fucus and upon their Mutual Influence in the Determination of the Developmental Axis 294
COE, WESLEY R.
Spermatogenesis in the California Oyster (Ostrea lurida) .... 309
BLUM. H. F., AND G. C. Me -BRIDE
Studies of Photodynamic Action, III. The difference in mech- anism between photodynamic hemolysis and hemolysis by non-irradiated cosine 316
CAROTHERS, E. ELEANOR
The Maturation Divisions and Segregation of Heteromorphic Homologous Chromosomes in Acrididae (Orthoptera) 324
ADOLPM, EDWARD F.
The Size of the Body and the Size of the Environment in the Growth of Tadpoles 350
ADOLPH, EDWARD F.
Body Size as a Factor in the Metamorphosis of Tadpoles. . . 376
JAHN, THEO. L.
Studies on the Physiology of the Euglenoid Flagellates, III. The effect of hydrogen ion concentration on the growth of Euglena gracilis Klebs 387
HALL, VICTOR E.
The Muscular Activity and Oxygen Consumption of Urechis
• aupo 400
CONTENTS v
BAUMBERGER, J. P., AND L. MIOIAKLIS
The Blood Pigments of Urechis caupo 417
FLORKIN, MARCEL, AND ALFRED C. REDFIELD
On the Respiratory Function of the Blood of the Sea Lion. . . 422
ROOT, R. W.
The Respiratory Function of the Blood of Marine Fishes. ... 427
HALL, F. G.
The Respiration of Puffer Fish 457
TANG, Pi-; i -SUNG
The Rate of Oxygen Consumption of Asterias Eggs before and after Fertilization • • 468
FAULKNER, G. H.
Notes on the Feeding Mechanism and on Intestinal Respira- tion in Chaetopterus variopedatus • • 472
WHITING, P. W.
Diploid Male Parts in Gynandromorphs of Habrobracon . . . 478
WHITING, P. W., AND M. F. STANCATI
A Gynandromorph of Habrobracon from a Post-reduced Bi- nucleate Egg 481
WILLIAMS, MARY MORRISON, AND M. H. JACOBS
On Certain Physiological Differences between Different Prepa- rations of So-Called "Chemically Pure" Sodium Chloride. . . 485
WELSH, JOHN H.
Specific Influence of the Host on the Light Responses of Parasitic Water Mites 497
PARPART, ARTHUR K.
Is Osmotic Hemolysis an All-or-None Phenomenon? 500
PARPART, A. K., W. R. AMBERSON AND D. R. STEWART
The Determination of Hemoglobin Concentration in Dilute Solutions v 518
Vol. LXI, No. 1 August, 1931
THE
BIOLOGICAL BULLETIN
PUBLISHED BY THE MARINE BIOLOGICAL LABORATORY
THE MARINE BIOLOGICAL LABORATORY
THIRTY-THIRD REPORT FOR THE YEAR 1930— FORTY-THIRD YEAR
I. TRUSTEES AND EXECUTIVE COMMITTEE (AS OK AUGUST 12,
1930) 1
LIBRARY COMMITTEE 3
II. ACT OF INCORPORATION 3
III. BY-LAWS OF THE CORPORATION 3
IV. REPORT OF THE TREASURER 5
V. REPORT OF THE LIBRARIAN 9
VI. REPORT OF THE DIRECTOR 10
Statement 10
Addenda :
1. The Staff, 1930 15
2. Investigators and Students, 1930 17
3. Tabular View of Attendance 28
4. Subscribing and Cooperating Institutions. 1930 .... 28
5. Evening Lectures, 1930 29
6. Shorter Scientific Papers, 1930 31
7. Members of the Corporation 34
I. TRUSTEES
EX OFFICIO
FRANK R. LILLIE, President of the Corporation, The University of Chicago. MERKEL H. JACOBS, Director, University of Pennsylvania. LAWRASON RIGGS, JR., Treasurer, 25 Broad Street, New York City. GARY N. CALKINS, Clerk of the Corporation, and Seer clary of the Board of Trustees, Columbia University.
EMERITUS
CORNELIA M. CLAPP, Mount Holyoke College.
C. R. CRANE, New York City.
H. H. DONALDSON, Wistar Institute of Anatomy and Biology.
OILMAN A. DREW, Eagle Lake, Florida.
WILLIAM PATTEN, Dartmouth College.
W. B. SCOTT, Princeton University.
E. B. WILSON, Columbia University.
1 1
2 MARIN'E BIOLOGICAL LABORATORY
TO SERVK UXTIL l'>34
E. R. CLARK, University of Pennsylvanin. E. G. CONKLIN, Princeton University. OTTO C. GLASER, Amherst College. Ross G. HARRISON. Yale University.
E. N. HARVEY, Princeton University.
H. S. JENNINGS, Johns Hopkins University.
F. P. KNOWLTOX, Syracuse University.
M. M. METCALF, Johns Hopkins University.
TO SERVE UNTIL 1933
H. C. BRADLEY, University of Wisconsin. I. F. LEWIS, University of Virginia. R. S. LILLIE, The University of Chicago. E. P. LYON, University of Minnesota.
C. E. McCLUNG, University of Pennsylvania.
T. H. MORGAN, California Institute of Technology. A. C. REDFIELD, Harvard University Medical School.
D. H. TENXENT, Bryn Maur College.
TO SERVF. rxTii. 1032
R. CHAMBERS, Washington Square College. Xc\v York University. \Y. E. GARREY, Vanclerhilt University Medical School. CASWELL GRAVE. Washington University.
M. J. GREENMAN. Wistar Institute of Anatomy and Biology. R. A. HARPER. Columbia University.
A. P. MATHEWS, The University of Cincinnati.
G. H. PARKER, Harvard University.
C. R. STOCKARD, Cornell University Medical College.
TO SERVE UNTIL 1931
H. C. BUMPUS. Broun University. W. C. CURTIS, University of Missouri.
B. M. DUGGAR, University <>f Wi-con^in.
GEORGE T. MOORE. Missouri Botanical Garden. St. Louis.
W. J. V. OSTERIIOUT, Member of the Rockefeller Institute for Medical
Research.
J. R. SCIIRAMM. University of Pennsylvania.
WILLIAM M. WHEELER. Bussev Institution. Harvard University. LORANDE L. WOODRUFF, Yale University.
EXECUTIVE COMMITTEE OF THE BOARD OF TRUSTEES
FRANK I\. LILLIE, R.\- Off. Chairman. MERKEL H. JACOBS, E.r. Off. LAWRASON RIGGS, JR., l:..v. Off. G. N. CALKINS, to serve until l'>31. L. L. WOODRUFF, to serve until 1931. W. C. CCKTIS, to serve until 1(>32. A. C. REDFIELD. to serve until 1(>32.
ACT OF INCORPORATION
THE LIBRARY COMMITTEE
C. E. McCLUNG, Chairman.
ROBERT A. BUDINGTON.
E. E. JUST.
M. M. METCALF.
ALFRED C. RF.DFIELD.
A. H. STURTEVANT.
w*wv
^S.-srfe.O.
No. 3170
II. ACT OF INCORPORATION COMMONWEALTH OF MASSACHUSETTS
Be It Known, That whereas Alpheus Hyatt. William San ford Stevens, William T. Sedgwick, Edward G. Gardiner, Susan Minns, Charles Sedg- wick Minot, Samuel Wells, William G. Farlmv, Anna D. Phillips and B. H. Van Vleck have associated themselves with the intention of forming a Corporation under the name of the Marine Biological Laboratory, for the purpose of establishing and maintaining a laboratory or station for scien- tific study and investigation, and a school for instruction in biology and natural history, and have complied with the provisions of the statutes of this Commonwealth in such case made and provided, as appears from the cer- tificate of the President, Treasurer, and Trustees of said Corporation, duly approved by the Commissioner of Corporations, and recorded in this office;
Now, therefore, I, HENRY B. PIERCE, Secretary of the Commonwealth of Massachusetts, do hcreb\ certify that said A. Hyatt, W. S. Stevens, W. T. Sedgwick, E. G. Gardiner. S. Minns, C. S. Minot. S. Wells, W. G. Farlow, A. D. Phillips, and B. H. Van Vleck, their associates and suc- cessors, are legally organized and established as, and are hereby made, an existing Corporation, under the name of the MARINE BIOLOGICAL LABORATORY, with the powers, rights, and privileges, and subject to the limitations, duties, and restrictions, which by law appertain thereto.
Witness my official signature hereunto subscribed, and the seal of the Commonwealth of Massachusetts hereunto affixed, this twentieth day of March, in the year of our Lord One Thousand Eight Hundred and Eighty- Eight. [SEAL]
HENRY B. PIERCE, Secretary of the Commonwealth.
III. BY-LAWS OF THE CORPORATION OF THE MARINE BIOLOGICAL LABORATORY
I. The annual meeting of the members shall be held on the second Tuesday in August, at the Laboratory, in Woods Hole, Mass., at 12 o'clock noon, in each year, and at such meeting the members shall choose by ballot a Treasurer and a Clerk to serve one year, and eight Trustees to serve four years. There shall be thirty-two Trustees thus chosen divided into four classes, each to serve four vears, and in addition there shall be two
4 MARINE BIOLOGICAL LABORATORY
groups of Trustees as follows: (a) Trustees ex officio, who shall be the President of the Corporation, the Director of the Laboratory, the Associate Director, the Treasurer and the Clerk; ( b} Trustees Emeritus, who shall be elected from the Trustees by the Corporation. Any regular Trustee who has attained the age of seventy years shall continue to serve as Trustee until the next annual meeting of the Corporation, whereupon his office as regular Trustee shall become vacant and be filled by election by the Cor- poration and he shall become eligible for election as Trustee Emeritus for life. The Trustees i\r officio and Emeritus shall each have the same right to vote as the regular Trustees.
The Trustees and officers shall hold their respective offices until their successors are chosen and have qualified in their stead.
II. Special meetings of the members may be called by the Trustees to be held in Boston or in Woods Hole at such time and place as may be designated.
III. The Clerk shall give notice of meetings of the members by pub- lication in some daily newspaper published in Boston at least fifteen days before such meeting, and in case of a special meeting the notice shall state the purpose for which it is called.
IV. Twenty-five members shall constitute a quorum at any meeting.
V. The Trustees shall have the control and management of the affairs of the Corporation; they shall present a report of its condition at every annual meeting; they shall elect one of their number President of the Cor- poration who shall also be Chairman of the Board of Trustees; they shall appoint a Director of the Laboratory ; and they may choose such other officers and agents as they may think best; they may fix the compensation and define the duties of all the officers and agents; and may remove them, or any of them, except those chosen by the members, at any time ; they may fill vacancies occurring in any manner in their own number or in any of the offices. They shall from time to time elect members to the Corporation upon such terms and conditions as they may think best.
VI. Meetings of the Trustees shall be called by the President, or by any two Trustees, and the Secretary shall give notice thereof by written or printed notice sent to each Trustee by mail, postpaid. Seven Trustees shall constitute a quorum for the transaction of business. The Board of Trustees shall have power to choose an Executive Committee from their own number, and to delegate to such Committee such of their own powers as they may deem expedient.
VII. The accounts of the Treasurer shall he audited annually by a certified public accountant.
VIII. The consent of every Trustee shall be necessary to dissolution of the Marine Biological Laboratory. In case of dissolution, the property shall be disposed of in such manner and upon such terms as shall be de- termined by the affirmative vote of two-thirds of the Board of Trustees.
IX. These By-laws may be altered at any meeting of the Trustees, pro- vided that the notice of such meeting shall state that an alteration of the By-laws will be acted upon.
X. Any member in good standing may vote at any meeting, either in person or by proxy duly executed.
REPORT OF THE TREASURER 5
IV. THE REPORT OF THE TREASURER To THE TRUSTEES OK THE MARINE BIOLOGICAL LABORATORY :
Gentlemen: Herewith is submitted my report as Treasurer of the Marine Biological Laboratory for the year 1930.
The accounts have been audited by Seamans, Stetson and Tuttle, certified public accountants. A copy of their report is on file at the Laboratory and is open to inspection by members of the Corporation.
At the end of the year 1930, the book value of the General Endow- ment Fund in the hands of the Central Hanover Bank and Trust Com- pany (of New York) as Trustee was $908,915 in securities and $34.50 in cash. The actual market value of the securities in this fund on the 9th day of May taking the mortgages at face value, was $931,981.25, showing a very satisfactory appreciation of the value over cost.
The book value of the Library Fund was $199,922.50 in securities and $77.50 in cash. The actual market value of the securities on May 9th was $203, 03 1.25.
At the end of the year the Lucretia Crocker Fund consisted of securi- ties of the book value of $4,707.59 and $374.32 in cash.
The Bio Club Scholarship Fund consisted of a mortgage participation of $2,000 and cash of $31.28, the Reynold A. Spaeth Memorial Lecture Fund of $3,000 in mortgage securities and cash of $75.46.
The Reserve Fund, consisting of the proceeds of the sale of part of the Bar Neck property to the Woods Hole Oceanographic Institution, consisted at the end of the year of bonds of the book value of $20,868.75 and cash of $3,090.55, of which cash $3,000 was later paid out under the contract, leaving net proceeds of the transaction of $20,959.30 which is being held with its income to meet maturing mortgage obligations or for such other purposes as the Trustees may decide.
The Retirement Fund at the end of the year consisted of $15,800 in- vested in mortgage participations, less an overdraft of $9.73, leaving $15,790.27.
The land, buildings, equipment and library, excluding the Devil's Lane and Gansett property, represented an investment of $1,617,086.71, less depreciation of $246,625.64, or a net amount of $1,370,461.07.
Current expenses including depreciation exceeded income for the year by $3,767.25.
Over $29,000 was expended from current funds on buildings, equip- ment and library.
At the end of the year the Laboratory owed $1,640.99 on accounts payable and $27,000 on bonds secured by mortgage.
Following is the Balance Sheet as of December 31, 1930, and the condensed statement of income and outgo for the year, also the surplus account.
6 MARINE BIOLOGICAL LABORATORY
EXHIBIT A
MARINE BIOLOGICAL LABORATORY BALANCE SHEET, DECEMBER 31, 1930
Assets
Endowment Assets and Equities :
Securities and Cash in Hands of Central Hanover Bank & Trust Company (of New York) Trustee— Schedules I-a and I-b $1,108,949.50
Securities and Cash — Minor Funds- Schedule II . 10.188.65 $1,119.138.15
Plant Assets :
Land— Schedule IV $ 97,103.05
Buildings— Schedule IV 1.207.354.03
Equipment— Schedule IV 155,222.28
Library— Schedule IV 157.407.35 $1. (.17.086.71
Less Reserve for Depreciation 246,625.64
$1.370.461.07
Securities and Cash in Reserve Fund 23.675.43
Cash in Dormitory Buildinu Fund SIS.% $1,394.955.46
Current Assets :
Cash ? 18.010.39
Accounts — Receivable 18.902.69
Inventories :
Supply Department $ 29.063.54
Bulletin 7,951.85 37.015.39
Investments :
Devil's Lane Property $ 37.780.91
Gansett Property 2.273.34
Stock in General Biological
Supply IlmiM'. Inc 12700.00
Retirement Fund Assets . 15.790.27 68.544.52
Prepaid Insurance 3.992.51 $146.465.50
Liabilities
Endowment Funds :
General Endowment Funds— Schedule III $1.108.949.50
Minor Endowment Funds— Schedule III 10.188.65 $1.119.138.15
Plant Funds :
Donations and Gifts— Schedule III $1.025.548.61
Other Investments in Plant from Gifts and Cur- rent Funds 364.406.85
$1,389.955.46
Mortgage. Danchakoff Estate 2.000.00
Accrued Charges on Sale of Bar Neck Land 3.000.00 $1.394.955.46
REPORT OF THE TREASURER
Current Liabilities and Surplus :
Mortgage, Devil's Lane Property $ 25,000.00
Accounts— Payable 1,640.99
Woods Hole Oceanographic Institution: Amount received for Purchase of
Books for their Library $2,500.00
Less Expenditures 2,147.07 352.93
Items in Suspense ( Net) 70.49
$ 27,064.41 Current Surplus— Exhibit C 119,401.09 $146,465.50
EXHIBIT B
MARINE BIOLOGICAL LABORATORY INCOME AND EXPENSE. YEAR ENDED DECEMBER 31, 1930
Total Net
Expense Income Expense Income Income : General Endowment Fund .... $ 48,020.46 $ 48,020.46
Library Fund 9,270.24 9,270.24
Gifts 500.00 500.00
Instruction 8.110.03 10,230.00 2,119.97
Research 4.069.37 16,261.06 12,191.69
Evening Lectures 135.48 135.48
Biological Bulletin and Member- ship Dues 7.557.17 9,421.18 1,864.01
Supply Department- Schedule V 62.030.00 62,162.82 132.82
Mess— Schedule VI 30.943.36 32,973.28 2,029.92
Dormitories —
Schedule VII 31.188.42 13,764.28 17,424.14
( Interest and Depreciation charged to above three De- partments. See Schedules
V. VI. and VII ) 35,424.79 35,424.79
Dividends, General Biological
Supply House. Inc 2,540.00 2,540.00
Rent, Danchakoff Cottages .... 634.11 1,039.00 404.89
Rent, Microscopes 462.50 462.50
Rent, Garage, Railway, etc. ... 154.90 154.00
Rait, Newman Cottage 137.27 150.00 12.73
Rent, Janitor's House 35.84 422.50 386.66
Sale of Duplicate Library Sets 2,198.13 2.198.13
Interest on Bank Balances" 529.87 529.87
Sundry Items 10.64 10.64
Maintenance of Plant :
New Laboratory Expense 16,839.26 16,839.26
Chemical and Special Appa- ratus 10.783.01 10,783.01
Maintenance, Buildings and Grounds 9,892.82 9.892.82
MARINE BIOLOGICAL LABORATORY
Library Department Expenses 8,912.66 8,912.66 Carpenter Department Ex- penses 1,516.91 1,516.91
Truck Expenses 851.48 851.48
Sundry Expenses 772.69 772.69
Bar Neck Property Expenses- 162.54 162.54 Workmen's Compensation In- surance 592.59 592.59
General Expenses :
Administration Expenses .... 14.509.68 14.509.68
Endowment Fund Trustee . . . 787.50 787.50
Interest on Loans 120.00 120.00
Bad Debts 317.98 317.98
Naples Zoological Station, for
Research 250.00 250.00
Mosquito Fund Contribution 100.00 100.00
Reserve for Depreciation 38,052.73 38.052.73
Excess of Expenses over Income carried to Current Surplus- Exhibit C 3.767.25 3767.25
$213,878.11 $213,878.11 $122,021.47 $122.021.47
EXHIBIT C
MARINE BIOLOGICAL LABORATORY, CURRENT SURPLUS ACCOUNT YEAR KNMKD DECEMBER 31, 1930
Balance, January 1. 1930 $119,933.29
Add:
Reserve for Depreciation charged to Plant Funds 38 052.73
Income from Retirement Fund 603.30
Cash received from Sale of Plant Assets deposited in Current
Cash (Motor-Boat) 50.00
$158.839.32 Deduct:
Payments from Current Funds during Year for Plant Assets as shown in Schedule IV,
Buildings $ 479.17
Equipment 5.464.23
Library Books, etc 23,099.38
$29;042.78
Purchase of Books from Balance of General Educa- tion Board Gift of $50,000.00 for Purchase of Books 5.708.20
Payment of Pensions from Retirement Fund 720.0C
Excess of Expenses over Income for Year as shown
in Exhibit B . 3,767.25 39,238.23
Balance. December 31, 1930— Exhibit A $119.401.09
Respectfully submitted,
LAWRASON RIGGS. JR..
Treasurer.
KKI'ORT OF TIIK LIBRARIAN
V. THE KKI'ORT OF THE LIBRARIAN
The important feature of W30, \vhich was the establishment of a regular endowment fund for the Library which would ordinarily give, along with the usual laboratory allowance, about $24,000 annually, was included in the report of last year. A general statement of the future apportionment of this sum as there given has been carried out in fact. A very important addition occurred in the spring, however, when the , Director of the Woods Hole Oceanographic Institution placed $5,000 at the disposal of the Library to be used exclusively for the purchase of oceanographic books and journals. Of this amount, $2,149.73 had been spent at the end of the year 1930. The items thus purchased are indi- cated specifically in the general statement of additions to the Library as follows : journal subscriptions were 346, 24 new, and of these 5 were for the Woods Hole Oceanographic Institution. One hundred and fif- teen books were purchased, 45 for oceanography. Back sets of journals were filled in complete to the number of 45, and 37 only partially com- pleted— none of these were regarded as strictly for the Woods Hole Oceanographic Institution. The number of journals received in ex- change for the Biological Bulletin was 442, an increase of 22, and 15 back sets that we needed were filled in. The reprint collection was augmented by 5,573.
The Library consists, then, of 26,519 bound journal volumes, 4,991 books, 64,231 reprints; and is receiving 1,060 current journals.
Gifts of books have been made to the Library by the following pub- lishers, and the Librarian takes this opportunity to acknowledge these in the name of the Marine Biological Laboratory Library, although formal thanks have in all cases been directly addressed by letter.
P. Blakiston's Son & Co 9
R. R. Bowker Co 1
Chemical Foundation, Inc 1
Chicago University Press 5
Harvard University Press 2
Paul B. Hoeber 2
Henry Holt & Co 1
Alfred A. Knopf 2
J. B. Lippincott Co 1
McGraw-Hill Book Co., Inc 5
Macmillan Co 21
C. V. Mosby Co 1
W. B. Saunders Co 4
Wm. Wood & Co 2
Yale University Press 1
10 MARINE BIOLOGICAL LABORATORY
VI. THE REPORT OF THE DIRECTOR To THE TRUSTEES OF THE MARINE BIOLOGICAL LABORATORY :
Gentlemen: I beg to submit herewith a report of the forty-third ses- sion of the Marine Biological Laboratory for the year 1930.
1. Attendance. The attendance for 1930 showed a slight increase over that of the preceding year in the numbers of both investigators and students, the figures for 1930 being 337 investigators and 136 students as compared with 329 investigators and 125 students in 1(>29. An in- spection of the Tabular View of .Attendance on page 28 will show ihat since 1927. when for the first time the research rooms in both the brick and the wooden buildings were practically all in use at the same time, the number of independent investigators has scarcely changed, except for the record-breaking summer of 1929 when visiting foreign physi- ologists, most of whom came to the Laboratory after the crowded sea- son, swelled the total to figures not likely to lit1 reached under normal conditions. On the other hand, investigators under instruction and re- search assistants whose numbers are not limited by that of the smaller laboratories have increased from 85 to 120 in the same period. The limit for the further increase of this class of investigators, however, is now in sight, and at the present rate will soon be reached.
The number of students, being limited by action of the Trustees, has shown only minor fluctuations for many years. The slight falling-off in 1929, caused chiefly by the change in that year in the times for holding the courses, was more than made up in 1930, though the maximum num- ber which may at present be admitted to the courses, namely 142. has not yet been reached. This failure of the registration to reach its maxi- mum value is not due to a deficiency of applications, since in nearly all of the courses the number of applicants greatly exceeds the number of available places, but rather to late withdrawals of students who have been accepted. To discourage such withdrawals, which are unfair to the rest of the applicants, who have usually in the meantime made other ar- rangements for the summer, the Executive Committee has recently voted to make a substantial increase in the registration fee which is forfeited in case of withdrawal.
Following the custom of the past three years, there are here pre- sented figures which show the distribution of the attendance of investi- gators throughout the four seasons, including that of 1930. for which the necessary records have been kept.
REPORT OF THE DIRECTOR
11
|
1927 |
1928 |
1929 |
1930 |
||
|
May |
30 |
7 |
15 |
9 |
6 |
|
June |
10 |
50 |
64 |
55 |
50 |
|
• • |
20 |
' 114 |
140 |
139 |
153 |
|
ft |
30 |
?] •> |
240 |
197 |
208 |
|
lulv |
10 |
247 |
281 |
238 |
253 |
|
• • |
20 |
247 |
282 |
242 |
250 |
|
a |
30 |
245 |
272 |
249 |
253 |
|
August |
10 |
234 |
250 |
256 |
254 |
|
ti |
20 |
208 |
226 |
243 |
245 |
|
.4 |
30 |
168 |
183 |
220 |
204 |
|
September |
10 |
110 |
112 |
157 |
122 |
|
tt |
20 |
50 |
43 |
59 |
44 |
|
,i |
30 . |
12 |
14 |
14 |
8 |
ujLIBRAKY
2. The Report of the Treasurer. This report shows that the total assets of the Laboratory at the end of 1930 were $2.660.559.11 as com- pared with $2,660,478.82 at the end of 1929. A further analysis of the figures shows that the hook value of the endowment fund has remained practically stationary (though the Treasurer calls attention to a grati- fying appreciation in the market value of the securities represented) while additions to the plant assets, chiefly in the form of hooks and new equipment, have about balanced the estimated depreciation on buildings and equipment. A decrease in the value of the land held by the Lab- oratory and the simultaneous appearance of a reserve fund of approxi- mately twenty thousand dollars are accounted for by the sale by the Laboratory to the Woods Hole Oceanographic Institution of the tract of land upon which the new building of the latter institution now stands.
The income of the Laboratory increased from $200,408.91 in 1929 to $210,110.86 in 1930. A part of this increase is of a temporary na- ture only, as for example that from the sale of duplicate sets by the library; of the remainder the largest single item is the additional in- come from the funds appropriated last year by the General Education Board. In 1930 for the first time the full annual income from this fund became available. Since, however, this fund is at present being ap- plied exclusively to the support of the library, the income available for general laboratory purposes remains practically unchanged.
The ordinary expenses of the Laboratory during 1930 showed a comparatively small increase over those for 1929, but expenses incidental to the reorganization of the Supply Department and the creation of a Museum, and, in particular, the reduction of the inventory of the Supply Department by discarding material originally valued at several thousand dollars, but for which there is at present little or no sale, have again pre- vented the appearance of a favorable balance, the excess of expenses over income after making allowance for depreciation being $3,767.25 as
12 MARINE BIOLOGICAL LABORATORY
compared with the similar deficit on paper of $855.33 for 1929. It is evident, however, when account is taken of the unusual expenses of the Supply Department in 1930 and of the necessarily large allowances for depreciation, that the finances of the Laboratory are in a very sound condition.
In 1930, for the first time in many years, the sum paid to the Lab- oratory for research space, chiefly by subscribing and cooperating insti- tutions, showed a slight decrease. This was to have been expected in view of business conditions, which have materially reduced the incomes of most of the institutions concerned. It is a very encouraging fact, however, that the decrease even under these abnormal conditions amounted to less than three per cent.
3. The Report of the Librarian shows a continuation of the steady growth of the library which has been made possible especially by the generous support of the General Education Board. For purposes of comparison the figures for 1930 may be added to those listed in the Director's Report for 1929.
1925 1926 1927 192S 1929 1930
Serials received currently 500 628 764 S74 985 1060 Total number of bound
volumes 15000 18200 22800 2o50i> 28300 31500
Reprints 25000 38000 43000 51000 59000 64000
One especially noteworthy feature of the past year lias been the strength- ening of the part of the library devoted to the subject of oceanography, the development in this direction having been made possible by a co- operative arrangement with the Woods Hole Oceanographic Institution, assisted by special funds appropriated by that institution.
4. Lectures and Scientific Meetings. During the season of 1930 the number of formal scientific lectures, including the Reynold A. Spaeth Memorial Lecture delivered by Professor Hardolph Wastenys of the University of Toronto, was thirteen, with several other evenings devoted to non-scientific lectures and motion pictures. In addition, there were held 13 less formal meetings at which 56 shorter papers, whose titles are given on pages 31 to 33, were presented and discussed. Two of these meetings were of especial interest. The first, held on June 27, assumed the character of a celebration of the sixtieth birthday of Dr. Frank R. Lillie and of the fortieth consecutive year of his con- nection with the Marine Biological Laboratory. In addition to the scientific papers presented on that occason. which were all based upon work having its inception in Dr. Lillie's laboratory, a special address of congratulation was delivered by 1 )r. I*".. 15. Wilson, and a ship's clock, the gift of Dr. Lillie's former students, was presented to him by Dr. L. Y.
REPORT OF THK DIRECTOR
Heilbrunn. The evening was concluded by an informal reception at the M. B. L. Club. The second meeting of an unusual character was the one held on the morning of July 26 at which 12 papers in the field of neuro-muscular physiology were presented and discussed. This meet- ing formed the most important part of a two days' program, social as well as scientific in character, which was arranged by the workers in this field and was attended not only by laboratory workers but by a number of physiologists from a distance. So successful was this meeting that it is to be hoped that similar ones, devoted to various fields of biological research may be held in the future.
5. Supply Department and Museum. At the last annual meeting of the Board of Trustees it was voted to develop for the use of investi- gators and students working in Woods Hole a museum in which speci- mens of the local fauna and flora may at all times be available for pur- poses of study, and in which there may at the same time be preserved full records of the distribution of all the local forms, the seasons of their maximum abundance, their breeding habits, etc. The development of this important activity of the Laboratory was very appropriately placed in the hands of Mr. George M. Gray, whose long experience as Curator of the Supply Department has given him unique qualification for such a position. In order to fill the vacancy thus created in the Supply De- partment and to provide for a possible ultimate separation of the two present functions of this Department, namely, that of supplying living material for experimental purposes to workers at the Laboratory and of furnishing preserved material to schools and colleges, the General Bio- logical Supply House of Chicago was invited to assume its temporary management. In preparation for the new arrangement, Dr. D. L. Gamble, representing this firm, spent several months in residence in Woods Hole during the summer of 1930 and has since continued the general supervision of this Department from Chicago with very satis- factory results, being ably assisted by Mr. James Mclnnis as Resident Manager.
6. Facilities for Work witJi X-rays. During the summer of 1930, through an appropriation of $500.00 by the Committee on the Effects of Radiation upon Living Organisms of the National Research Council and with the active assistance of the Chairman of that Committee, Pro- fessor W. C. Curtis, special facilities, not hitherto available for work with X-rays and other radiations were provided for workers at the Lab- oratory. In particular, there was made available throughout the summer the expert advice of Dr. G. Failla of the Memorial Hospital, New York, together with the assistance of competent technicians in the operation of the apparatus. Several manufacturers also furnished very valuable aid
14 MARIXE BIOLOGICAL LABORATORY
of various sorts which is here gratefully acknowledged. So successful was this arrangement in 1930 that it is gratifying to he ahle to announce at the time of the writing of this report that it will he continued and somewhat extended in 1931.
7. The Woods Hole Oceanographic Institution. The position which Woods Hole has held for many years as one of the leading hiological centers in the United States was materially strengthened hy the erection during the past year of the large and splendidly equipped building which will be the permanent headquarters of the \Yoods Hole Oceanographic Institution. The land on which this building stands was formerly held by the Marine Biological Laboratory and leased by it to the liar Xeck Corporation. At a special meeting of the Board of Trustees, held in Washington. I). C., on April 28, 1930. it was voted to enter into an agreement with the Woods Hole Oceanographic Institution and the liar Neck Corporation by which the former institution would acquire by purchase1 from the Marine Biological Laboratory approximately 54,000 square feet of the westerly portion of the so-called " Bar Xeck Wharf." Full details as to this agreement will be found in the Re- port of the Auditors for 1930. Though there is no official connection between the Woods Hole Oceanographic Institution and the Marine Biological Laboratory, the work of each institution will supplement that of the other, and it is planned that there shall be close scientific co- operation between them. This cooperation has already assumed the form of a sharing of library facilities and Mess accommodations and will be extended in the future in all possible ways.
8. The IJoanl of Trustees. ( )ne change in the Board of Trustees occurred during the past year, Professor William Patten of Dartmouth College having been elected Trustee Emeritus at the annual meeting of the Corporation and Professor E. R. Clark of the Cniversity of Penn- sylvania having been selected to till tin- place thereby made vacant in the Class of 1934.
9. Gifts. Appreciative acknowledgment is made of the gift by Mr. Ware Cattell and the " Collecting Net " of $500.00 for scholarships to students who in the courses given bv the Laboratory show unusual promise as research workers.
There are appended as parts of this report :
1. The Staff, 1930.
2. Investigators and Students, 1930.
3. A Tabular View «>f Attendance. 1926-1930.
4. Subscribing and Cooperating Institutions, 1930.
5. Evening Lectures, 1930.
REPORT OF THE DIRECTOR
6. Shorter Scientific Papers. 1930.
7. Members of the Corporation, August, 1930.
Respectfully submitted,
M. H. JACOBS,
Director.
1. THE STAFF, 1930
MERKEL H. JACOBS. Director, Professor of General Physiology, University
of Pennsylvania. Associate Director: -
ZOOLOGY
I. IXVKSTIGATIOX
GARY N. CALKINS, Professor of Protozoology, Columbia University.
E. G. CONKLIN, Professor of Zoology, Princeton University.
CASWELL GRAVE, Professor of Zoology, Washington University.
H. S. JENNINGS, Professor of Zoology, Johns Hopkins University.
FRANK R. LILLIE, Professor of Embryology, University of Chicago.
C. E. McCLUNG, Professor of Zoology, University of Pennsylvania.
S. O. MAST, Professor of Zoology, Johns Hopkins University.
T. H. MORGAN, Director of the Biological Laboratory, California Institute
of Technology.
G. H. PARKER, Professor of Zoology, Harvard University. E. B. WILSON, Professor of Zoology, Columbia University. LORANDE L. WOODRUFF. Professor of Protozoology, Yale University.
II. INSTRUCTION
J. A. DAWSON, Assistant Professor of Biology, College of the City of New
York.
T. H. BISSONNETTE, Professor of Biology, Trinity College. E. C. COLE, Associate Professor of Biology, Williams College.
0. E. NELSEN, Instructor in Zoology, University of Pennsylvania. A. W. POLLISTER, Instructor in Zoology, Columbia University.
L. P. SAYLES, Instructor in Biology, College of the City of New York.
A. E. SEVERINGHAUS, Assistant Professor of Anatomy, College of Phy- sicians and Surgeons, Columbia University.
JUNIOR INSTRUCTORS
B. R. COONFIELD, Professor of Biology, Southwestern College.
1. B. HANSEN, Graduate Student, University of Chicago.
PROTOZOOLOGY
I. INVESTIGATION (See Zoology}
II. INSTRUCTION
GARY N. CALKINS, Professor of Protozoology, Columbia University. RACHEL BOWLING, Instructor in Zoology, Columbia University. ROBERTS RUGH, Assistant in Zoology. Columbia University.
16 MARINE BIOLOGICAL LABORATORY
EMBRYOLOGY
I. INVESTIGATION (Sec Zoology}
II. INSTRUCTION
HUBERT B. GOODRICH, Professor of Biology, Wesleyan University. BENJAMIN H. GRAVE, Professor of Biology, De Pamv University. CHARLES PACKARD, Assistant Professor of Zoology, Institute of Cancer
Research, Columbia University.
CHARLES G. ROGERS, Professor of Comparative Physiology, Oberlin College. B. C. T WITTY, Instructor in Zoology, Yale University.
PHYSIOLOGY
I. INVESTIGATION
HAROLD C. BRADLEY, Professor of Physiological Chemistry, University of Wisconsin.
WALTER E. CARREY, Professor of Physiology, Vanclerbilt University Med- ical School.
RALPH S. LILLIE, Professor of General Physiology, University of Chicago.
ALBERT P. MATHEWS, Professor of Biochemistry, University of Cincinnati.
II. INSTRUCTION Teaching Staff
WILLIAM R. AMBERSON, Assistant Professor of Physiology, University of Pennsylvania.
PHILIP BARD, Assistant Professor of Physiology, Princeton University.
HALLOWELL DAVIS, Assistant Professor of Physiology, Harvard University.
RALPH W. GERARD, Assistant Professor of Physiology, University of Chi- cago.
CHARLOTTE HAYWOOD, Assistant Professor of Physiology, Vassar College.
LEONOR MICHAELIS, Member of the Rockefeller Institute, New York City.
Special Lecturers
EDWIN J. COHN, Associate Professor of Physical Chemistry, Harvard Uni- versity.
HENRY J. FRY, Associate Professor of Biology, Washington Square College, New York University.
E. NEWTON HARVEY, Professor of Physiology, Princeton University.
SELIG HECHT, Professor of Biophysics, Columbia University.
MI-.RKEL H. JACOBS, Professor of General Physiology, University of Penn- sylvania.
BALDUIN LUCKE, Associate Professor of Pathology, University of Pennsyl- vania.
BOTANY
I. INVESTIGATION
B. M. DUGGAR, Professor of Physiological and Economic Botany, University of Wisconsin.
C. E. ALLEN, Professor of Botany, University of Wisconsin.
REPORT OF THE DIRECTOR 1 '
S. C. BROOKS, Professor of Zoology, University of California. IVEY F. LEWIS, Professor of Biology, University of Virginia. WM. J. ROBBINS, Professor of Botany, University of Missouri.
II. I: xs'i RUCTION
WILLIAM RANDOLPH TAYLOR, Professor of Botany, University of Penn- sylvania.
H. C. BOLD, Instructor in Botany, University of Vermont. JAMES P. POOLE, Professor of Evolution, Dartmouth College.
LIBRARY
PRISCILLA B. MON -IGOMKRY ( MRS. THOMAS H. MONTGOMERY, JR.), Li- brarian.
DEBORAH LAWRENCE. Secretary. HESTER ANN BRADBURY, LILLIAN F. BRIGGS, MARY A. ROHAN, Assistants.
CHEMICAL SUPPLIES
OLIVER S. STRONG. Professor of Xeurology and Xeuro-Histology. Columbia University, Chemist.
APPARATUS ROOM
SAMUEL E,. POND, Assistant Professor of Physiology, Medical School, Uni- versity of Pennsylvania, Custodian of Apparatus.
MUSEUM GEORGE M. GRAY, Curator.
SUPPLY DEPARTMENT
JAMES MC!NNIS, Manager. WALTER KAHLER, Collector.
A. M. HILTON, Collector. GEOFFREY LEHY, Collector.
MILTON B. GRAY, Collector. A. W. LEATHERS, Shipping.
BOATS JOHN J. VEEDER, Captain. E. M. LEWIS, Chief Engineer.
F. M. MACNAUGHT, Business Manager.
HERBERT A. HILTON, Superintendent of Buildings and Grounds.
THOMAS LARKIN. Superintendent of Mechanical Department.
LESTER F. Boss, Mechanician.
J. D. GRAHAM, Glass-blowing Service.
A. R. APGAR, Photographic Service.
WILLIAM HEMENWAY, Carpenter.
2. INVESTIGATORS AND STUDENTS, 1930 Independent Investigators
ABRAMSON, HAROLD A.. Instructor. Harvard University.
AMBERSON. WILLIAM R.. Assistant Professor of Physiology, University of Penn- sylvania. 2
18 MARINE BIOLOGICAL LABORATORY
ARMSTRONG, PHILIP B., Instructor in Anatomy, Cornell University Medical College.
ASHWORTH, JAMES H., Professor of Natural History, University of Edinburgh.
ASTROM, I. ELISABETH, Class Assistant, University of Toronto.
BAITSELL, GEORGE A., Professor of Biology, Yale University.
BAKER, HORACE B., Associate Professor, University of Pennsylvania.
BALL, ERIC G., National Research Fellow in Medicine, Johns Hopkins University Medical School.
BARD, PHILIP, Assistant Professor of Physiology, Princeton University.
BARRON, E. S. GUZMAN, Instructor in Medicine, Johns Hopkins University Medical School.
BARTH, L. G., National Research Fellow, University of Chicago.
BEAMS, H. W., Dupont Fellow, University of Virginia.
BELKIN, MORRIS, Instructor, New York University.
BIDDLE, RUSSELL L., Teaching Fellow. California Institute of Technology.
BISSONNETTE, T. HUME, Professor of Biology, Trinity College.
BLANCHARD, KENNETH C., Associate Professor, New York University.
BLUMENTHAL, REUBEN, Harrison I-Yllow in Zoology, University of Pennsylvania.
BOLD, HAROLD C.. Instructor in Botany. University of Vermont.
BORODIN, D. N., 621 West 142d Street, New York City, New York.
BOWLING, RACHEL, Instructor in Zoology, Columbia University.
BRAPWAV, WINNIFRED, New York University.
BREITENBECHER, J. K., McGill University.
BRIDGES, CALVIN B., Research Assistant, Carnegie Institution of Washington.
BRONFENBRENNER, J., Professor of Bacteriology, Washington University Medical School.
BRONK, DETLEV W.. Professor of Biophysics and Director of Johnson Foundation for Medical" Physics, University of Pennsylvania.
BROOKS, MATILDA M., Research Associate in Biology, University of California.
BROOKS, S. C., Professor of Zoology, University of California.
BURDICK, DONALD L., Instructor in Biology, Union C»lK-m -.
BYTINSKI-SALZ, HANS, Research Fellow, Yale University.
CALKINS, GARY N., Professor of Proto/m'ilogy, Columbia University.
CAROTHERS, E. ELEANOR, Lecturer in Zoology, University of Pennsylvania.
CATTELL, WARE, Research Fellow in Biophysics, Memorial Hospital.
CHALMERS, ELIZABETH, Graduate Assistant. University of Pittsburgh.
CHATTON, EDOUARD, University of Strasbourg, Strasbourg, France.
CHEEVER, CLARENCE A., Boston Society of Natural History, Boston, Mass.
CHIDESTER, FLOYD E., Professor of Zoology, West Virginia University.
CHOUKE, K. S., Assistant Professor of Anatomy. School of Medicine, University of Colorado.
CHRISTIE, JESSE R., Associate Nematologist, United States Department of Agri- culture.
CLOWES, G. H. A., Director, The Lilly Research Laboratory. Eli Lilly & Co.
COBB. N. A., Agricultural Technologist and Nematologist, United States Depart- ment of Agriculture.
COE, W. R., Professor of Biology, Yale University.
COKFEY, J. M., Assistant Bacteriologist, New York State Department of Health.
COLE, ELBF.RT C., Associate Professor of Biology, Williams College.
COLE, KENNETH S., Assistant Professor of Physiology, Columbia University.
COONFIELD, BENJAMIN R., Professor of Biology, Southwestern College.
COOPER, GEORGE O., Instructor, University of Wisconsin.
Coi'ELAND, MANTON, Professor of Biology, Bowdoin College.
COWLES, R. P., Associate Professor of Zodlogy, Johns Hopkins University.
CURTIS, W. C., Professor of Zoology, University of Missouri.
DAVIS, HALLOWELL, Assistant Professor of Physiology, Harvard University Med- ical School.
REPORT OF THE DIRECTOR 19
DAWSON, ALDEN B., Associate Professor of Zoology, Harvard University.
DAWSON, J. A., Assistant Professor of Biology, College of the City of New York.
DILL, D. B., Assistant Professor of Biochemistry, Harvard University.
DOLLEY. WILLIAM I.., JR., Professor of Biology, University of Buffalo.
Du Bois, DELAFIELD, Washington Square College, New York University.
Du Bois, EUGENE F., Associate Professor of Medicine, Cornell University Med- ical College.
DuBuissoN, MARCEL, Professor of Zoology, " Ecole des Hautes fitudes," Ghent, Belgium.
DUGGAR, B. M., Professor of Plant Physiology and Applied Botany, University of Wisconsin.
EDWARDS, DAYTON J., Associate Professor of Physiology, Cornell University Medical College.
EDWARDS. H. T., Assistant in Fatigue Laboratory, Harvard University.
FAILLA, G., Physicis-t, Memorial Hospital, New York.
FANKHAUSER, GERHARD, Fellow of the Rockefeller Foundation, University of Chi- cago.
FINLEY, HAROLD E., Instructor in Zoology, West Virginia State College.
FLORKIN, MARCEL, Research Fellow, Harvard University.
FREW, PRISCILLA E.. Instructor, Hunter College.
FRY, HENRY J., Associate Professor of Biology, Washington Square College, New York University.
FURTH, JACOB, Associate in Pathology, The Henry Phipps Institute, University of Pennsylvania.
GARDINER, MARY S., Associate in Biology, Bryn Mawr College.
GARREY, W. E., Professor of Physiology, Vanderbilt University Medical School.
GATES, FREDERICK L., Research Fellow, Harvard University.
GELFAN, SAMUEL, Research Fellow, University of Chicago.
GERARD, R. W., Assistant Professor of Physiology, University of Chicago.
GIBBONS, NORMAN E., Graduate Student, Yale University.
GOLDFORB, A. J., Professor of Biology, College of the City of New York.
GOODRICH, HUBERT B., Professor of Biology, Wesleyan University.
GRAVE, B. H., Professor of Zoology, DePauw University.
GRAVE, CASWELL, Professor of Zoology, Washington University.
GRUNDFEST, HARRY, National Research Council Fellow, Columbia University.
HANCE, ROBERT T., Head of Department of Zoology, University of Pittsburgh.
HARVEY, ETHEL B., Instructor, Washington Square College, New York University.
HARVEY, E. NEWTON, Professor of Physiology, Princeton University.
HAYWOOD, CHARLOTTE, Assistant Professor of Physiology, Vassar College.
HEILBRUNN, L. V., Associate Professor of Zoology, University of Pennsylvania.
HENSHAW, PAUL S., Biophysicist, Memorial Hospital.
HETLER. DONALD M., Instructor in Bacteriology, Washington University Medical School.
HIBBARD, HOPE, Assistant Professor, Oberlin College.
HILL, SAMUEL E., Assistant in Physiology, Rockefeller Institute.
HINRICHS. MARIE A., Research Associate in Physiology, University of Chicago.
HINTZE, A. LAURA, Assistant Professor of Physiology, Goucher College.
HIRSCH, G. C.. Professor of Zoology, University of Utrecht.
HOPPE, ELLA N., Research Assistant, New York State Department of Health.
HOWE, H. E., Editor, Industrial and Engineering Chemistry.
ROWLAND, RUTH B., Associate Professor of Biology, Washington Square College, New York University.
HUETTNER, ALFRED F., Associate Professor, Washington Square College, New York University.
HUGHES, THOMAS P., Associate in Bacteriology, Rockefeller Institute.
HUNTER, LILLIAN M., Graduate Student and Assistant Technician, University of Toronto.
20 MARINE BIOLOGICAL LABORATORY
HUTCHINSON, G. E., Instructor in Biology. Yale University.
HYMAN. LIBBIE H.. Research Associate, University of Chicago.
JACOBS, M. H., Professor of General Physiology-, University of Pennsylvania.
JENNINGS, H. S.. Professor of Zoology. Johns Hopkins University.
JOHLIN, J. M., Associate Professor of Biochemistry, Vanderbiit University Med- ical School.
JOHNSON, H. HERBERT, Instructor, College of the City of Xew York.
JUST, E. E., Professor of Zoology, Howard University.
KEIL, ELSA M.. Instructor in Zoology, Rutgers University.
KELTCH, ANNA K.. Research Chemist, Lilly Research Laboratory.
KETTLEKAMP, B. H., Instructor, University of Pittsburgh.
KEVES, D. B., Professor of Industrial Chemistry. University of Illinois.
KEYS. ANCEL B.. Fellow in the Biological Sciences, .National Research Council.
KNOWER. HENRY Me K.. Wistar Institute.
KNOWLTON. FRANK P.. Professor of Physiology, Syracuse University, College of Medicine.
KOEHRING, VKRA. Beaver College, Jenkintown, Pennsylvania.
KUNITZ, MOSES. Associate Professor. Rockefeller Institute for Medical Research.
LACKEY. J. B.. Professor of Biology. Southwestern University.
LANCEFIELD, D. K.. Associate Professor in Zoology, Columbia University.
LANCEFIELD, REBECCA C. Assistant in Bacteriology, Rockefeller Institute for Medi- cal Research.
LEWIS, IVEY F., Professor of Biology. University of Virginia.
LII.I.IE. FRANK R.. Chairman of the Department of Zoology, University of Chicago.
I. ii. i. IK, RALPH S., Professor of General Physiology, University of Chicago.
LUCKE, BALDUIN, Associate Professor of Pathology, University of Pennsylvania.
LYNCH. RUTH S.. Instructor in Graduate Zoology. The Johns Hopkins University.
LYON, E. P., Professor of Physics, University of Minnesota.
McCLUNG, C. E., Director, Zoological Laboratory, and Professor of Zoology, Uni- versity of Pennsylvania.
MAcDouGAi.i., MARY S.. Head of Biology Department. Agnes Scott College.
M< F.WF.N. ROBERT S.. Associate- Professor of Zoology. Oherlin College.
McGLONE. MARTI. is. Instructor in Physiology. University of Pennsylvania.
MANWEIX. RKGIXAI.D I).. Instructor, Johns Hopkins University.
MARSLANP, DOUGLAS A., Assistant Professor of Biology. Washington Square College, Xew York University.
MATHEWS, A. P.. Professor of Biochemistry. University of Cincinnati.
MATTHEWS. SAMUEI A., Instructor. University of Pennsylvania.
MAVOR, JAMF.S W., Professor of Biology and Head of Department. Union College.
MENKIN, VALY, Fellow in Medicine, Henry Phipps Institute. University of Pennsylvania.
MEYER. Roi \NI> K.. Research Assistant. University of Wisconsin.
MICHAELIS, LF.ONOR, Member, Rockefeller Institute for Medical Research.
MII.LEH. Hiii'.v M.. Fellow, National Research Council, Johns Hopkins University.
MITCHELL. Piiiiir H., Professor of Physiology. Brown University.
MONNIER, ANTIREE. University of Paris.
MONNIER, A. M.. Assistant at the Sorbonne. Paris.
MORGAN, LILIAN V.. California Institute of Technology.
MORGAN. T. H.. Director of the Biological Laboratory, California Institute of Technology.
MORGULIS, S.. Professor of Biochemistry. University of Xebraska. College of Medicine.
MORRILL, C. V., Associate Professor of Anatomy. Cornell University Medical College.
MORRIS. HEI.F.N S.. Graduate Student. Columbia University.
MULDER, ARTHUR G., Associate Professor of Physiology. University of Tenness-ee.
REPORT OF THE DIRECTOR
NABRIT, S. MILTON, Head of Department of Biology, Morehousc College.
NELSON, OLIN E., Instructor in Zoology, University of Pennsylvania.
NICHOLAS, WARREN W., X-Ray Physicist, National Bureau of Standards.
NONIDEZ, JOSE F., Assistant Professor of Anatomy, Cornell University Medical College.
PACKARD, CHARLES, Assistant Professor of Zoology, Columbia University.
PARK, ORLANDO, Associate Professor of Biology, Kent State College.
PARMENTER, CHARLES L., Associate Professor of Zoology, University of Penn- sylvania.
PAYNE, FERNANDAS, Professor of Zoology and Dean of Graduate School, Indiana University.
PERROT, J. L., Columbia University.
PETRIK, JOSEPH M., Director of the Department of Physiology, Masaryk Uni- versity.
PHILLIPS, PAUL L., Instructor in Anatomy, Cornell University Medical College.
PHILPOTT, CHARLES H., Lecturer in Medical Zoology, Washington University Medical School.
PLOUGH, HAROLD H., Professor of Biology, Amherst College.
POLLISTEK, ARTHUR W., Instructor in Zoology, Columbia University.
POND, SAMUEL E., Assistant Professor of Physiology, University of Penn- sylvania Medical School.
POOLE, JAMES P., Professor of Evolution, Dartmouth College.
POTTER, TRUMAN S., Seymour Coman Fellow, University of Chicago.
RAFFEL, DANIEL, National Research Fellow, Johns Hopkins University.
REDFIELD, HELEN, California Institute of Technology.
REESE, ALBERT M., Head of Department of Zoology, West Virginia University.
RICHARDS. OSCAR W.. Assistant Professor of Biology, Clark University.
ROBERTS, EDITH A.. Chairman of Department of Botany, Vassar College.
ROGERS, CHARLES G., Professor of Comparative Physiology, Oberlin College.
ROMANOFF, ALEXIS L., Research Instructor, Cornell University.
ROOT, \VALTER S., Assistant Professor, Syracuse University Medical School.
RUGH, ROBERTS, Assistant in Zoology, Columbia University.
SAYLES, LEONARD P., Instructor in Zoology, Tufts College.
SCHAUFFLER, WILLIAM G., Princeton, New Jersey.
SCHMIDT, LEON H.. University of Cincinnati.
SCHMITT, FRANCIS O., Assistant Professor of Zoology, Washington University.
SCHRADER, FRANZ, Associate Professor, Bryn Mawr College.
SCHRADER, SALLY-HUGHES, Instructor in Biology, Bryn Mawr College.
SCHULTZ, JACK, Research Assistant, Carnegie Institution of Washington.
SEVERINGHAUS, AURA E.. Associate in Anatomy, Medical School, Columbia Uni- versity.
SHARMA, B. M., Professor of Anatomy, Tibbi Medical College.
SHOUP, CHARLES S., Assistant Professor of Biology, Vanderbilt University.
SHULL, A. FRANKLIN, Professor of Zoology, University of Michigan.
SHUMWAY, WALDO, Professor of Zoology, University of Illinois.
SICHEL, FERDINAND J., Assistant Instructor, Washington Square College, New York University.
SLIFER, ELEANOR H., Graduate Student. University of Pennsylvania.
SMITH, FREDERICK, Research Assistant, Rockefeller Institute.
STEINBACH, H. B., Graduate Student, Brown University.
STOCKARD, CHARLES R., Professor of Anatomy, Cornell University Medical College.
STRONG, OLIVER S., Professor of Neurology and Neuro-Histology, Columbia Uni- versity.
STUNKARD, H. W., Professor of Biology, New York University.
STURDIVANT, HARWELL P., Instructor, Columbia University.
STURTEVANT, A. H., Professor of Genetics, California Institute of Technology.
22 MARINE BIOLOGICAL LABORATORY
SUM WALT, MARGARET, Assistant Professor, Woman's Medical College.
TASHIRO. SHIRO, Professor of Biochemistry, The University of Cincinnati.
TAYLOR, WM. RANDOLPH, Professor of Botany, University of Pennsylvania.
THOMPSON, J. W., Instructor, Swarthmore College.
TOWER, SARAH S.. Instructor in Anatomy, Johns Hopkins University.
TWITTY, VICTOR C., Instructor in Zoology, Yale University.
UHLENHUTH, EDUARD, Professor of Gross Anatomy. University of Maryland Medical School.
VAN CLEAVE, C. D., Instructor in Anatomy. University of Pennsylvania.
VAN SLYKE, E., Instructor, University of Pittsburgh.
WANG, CHI CHE, 1314 East 56th Street, Chicago, Illinois.
WARREN, HOWARD C., Stuart Professor of Psychology, Princeton University.
WENRICH, D. H., Professor of Zoology, University of Pennsylvania.
WESSON, LAURENCE G., Assistant Professor of Pharmacology, Vanderbilt Univer- sity Medical School.
WIIKDON, ARTHUR D.. Professor of Zoology and Head of Department, North Dakota Agricultural College.
WHITAKER, D. M., Assistant Professor of Zoology, Columbia University.
WHITING, ANNA R., Professor, Head of Department of Biology, Pennsylvania College for Women.
WHITING, P. W., Associate Professor of Zoology, University of Pittsburgh.
WIEMAN, H. L., Professor of Zoology, University of Cincinnati.
WILLIER, B. H., Associate Professor of Zoology. University of Chicago.
WILSON, EDMUND B., DaCosta Professor of Zoology, Emeritus, Columbia Uni- versity.
WILSON, F. EDWARD, Graduate Student, Clark University.
WILSON, MAY G., Associate, Department of Pediatrics, Cornell University Medical College.
WOLF, E. ALFRED, Assistant Professor of Zoology. Unm-i-Miy of Pittsburgh.
WOODRUFF, L. L., Professor of Protozoology. Yale University.
WOODS, FARRIS H.. Assistant Profes>or of /mildly. University of Missouri.
YOUNG, WILLIAM C.. Instructor in Biology, Brown University.
ZELENY, CHARLES. Professor of Zoology, University of Illinois.
Beginning Investigators
BAILEY, SARAH Wr., Graduate Student. Radcliffe College.
BROWN, FRANK A., JR., Austin Teaching Fellow, Harvard University.
CALDWELL, LUCILE. Johns Hopkins University.
CHANG, J. H., Graduate Student. University of Chicago.
CHEN, H. T., Student, Harvard University Medical School.
CLINE, ELSIE, Graduate Student, The Johns Hopkins University.
COLDWATER, KENNETH B., Instructor in Zoology, University of Missouri.
COSTELLO, DONALD P., Assistant in Zoology, College of the City of Detroit.
CROASDALE, HANNAH T., Graduate Student, University of Pennsylvania.
CURTIS, MARY ELIZABETH, Graduate Student, Yale University.
DAMBACH, GEORGE J., Graduate Assistant, University of Pittsburgh.
DEARING, WILLIAM H., Graduate Student, University of Pennsylvania.
DEITRICH, JOHN E., Medical Student, Johns Hopkins University.
DIXON, EVELYN C., Graduate Student. Washington University.
DRAPER. JOHN W., Cornell University Medical College.
DREW, WILLIAM, Massachusetts Agricultural College.
DuSnANE, GRAHAM, Wabash College.
ETKIN, WILLIAM, Tutor. College of the City of New York.
FLAMMON. SISTER M. MURIEL. Instructor in Biology, Seton Hill College.
FRENCH, C. S., Harvard University.
GEBRAK, ANTON, Moscow Agricultural Academy.
KKPORT OF THE DIRECTOR
GEIB, DOROTHY A., Student, Johns Hopkins University Medical School.
GEIMAN, QUENTIN M., Graduate Student, University of Pennsylvania.
GENTHER, IDA T., Assistant in Zoology, Washington University.
GRAUBARD, MARC A., Assistant in Zoology, Columbia University.
GREEN, DAVID E., Assistant, New York University.
GUERLAC, HENRY E., Cornell University.
HANSEN, IRA B., Instructor, Wesleyan University.
HAYNES, FLORENCE W., 20 Gorham Road, West Medford, Massachusetts.
HEGNAUER. ALBERT, Assistant in Physiology, University of Rochester, School of Medicine.
HILEMAN, CLARA M., Teacher of Biology, Columbia University.
HOERR, STANLEY O., Antioch College.
HOOK, SABRA J., Assistant in Zoology, Barnard College, Columbia University.
IGLAUER, CHARLES A., Graduate Student, University of Pennsylvania.
IMLAH, HELEN- W., Graduate Student, Radcliffe College.
JEFFERY, HELEN, Research, Washington University Medical School.
KALTREIDER, NOLAN L., Student, Johnson Foundation, University of Pennsylvania.
KATZ, JACOB D., Assistant Instructor, Washington Square College, New York Uni- versity.
KERR, THOMAS, Instructor in Biology, New York University.
KIXNEY, ELIZABETH T., Assistant at Barnard College, Columbia University.
KINSBERGEN, MAURICE, Assistant, New York University.
KLOSE, THEODORA, Instructor in Botany, Vassar College.
LEE, KATY, Graduate Assistant in Zoology, University of Missouri.
LHERISSON, CAMILLE, Professor of Biology, University of Haiti Medical School.
LORBERBLATT, ISAAC, Student, Washington University Medical School.
McGouN, RALPH C, JR., Instructor in Biology, Amherst College.
MACMURRAY, MARY T., 8629 109th Street, Richmond Hill. New York.
MACKMULL, GULDEN, Demonstrator of Histology and Embryology, Baugh Insti- tute of Anatomy.
MILLER, EVELYN H., Graduate Student, University of Pennsylvania.
MILLER, FORREST W., Graduate Assistant, University of Pittsburgh.
MONAGHAN, BETTY R., Assistant, Washington University.
MORRIS, SAMUEL, Instructor in Zoology, University of Pennsylvania.
NELSON, PHYLLIS M., Washington University.
PARPART, ARTHUR K., Instructor and Graduate Student, University of -Pennsylvania.
PARPART, ETHEL R., Technician, University of Pennsylvania.
PITTS, ROBERT F., Student Assistant, Johns Hopkins University.
RANKIN, DOUGLAS, Johns Hopkins University.
ROBERT, NAN L., Instructor, Hunter College.
SANTOS, FELIX V., Graduate Student, The University of Chicago.
SAVIN, MARION B., Graduate Student, University of Pennsylvania.
SCARBOROUGH, J. ELLIOTT, JR., Student of Medicine, Harvard University.
SCHECHTER, VICTOR, Tutor in the Department of Biology, College of the City of New York.
SCHWEITZER, MORTON D., Assistant in Zoology, Columbia University.
SCOTT, SISTER FLORENCE M., Assistant Professor of Biology, Seton Hill College.
SHAPIRO, HERBERT, Assistant in Zoology, Columbia University.
SHAW, C. RUTH, Graduate Assistant, University of Pittsburgh.
SKINNER, B. F., Graduate Student, Harvard University.
SMITH, SUZANNE G., Graduate Assistant in Zoology, University of Missouri.
SMYTHE, C. V., Fellow National Research Council, Rockefeller Institute.
SONNEBORN, TRACY M., Research Assistant, Johns Hopkins University.
STABLER, ROBERT M., Assistant Instructor, University of Pennsylvania.
STANCATI, MILTON F., Graduate Assistant, University of Pittsburgh.
STEWART, DOROTHY R., Assistant Professor of Biology, Skidmore College.
24 MARINE BIOLOGICAL LABORATORY
STEINER, MATTHEW M., Assistant in Biology, New York University.
STREET, SIBYL F., Assistant to Department of Zoology, Yassar College.
STUCK, FLORENCE, Student, Columbia University.
TANG, PEI-SUNG, Johns Hopkins University.
TE\YINKEL, Lois E., Assistant in Zoology, Barnard College, Columbia University.
TOOTH ILL, MARTHA C.. Assistant in Biology, Brown University.
TUAN, Hsu-CnvAN. Graduate Student. University of Pennsylvania.
TURNER, CLARENCE D., Research Assistant, University of Missouri.
WATERS, NELSON F., Research Assistant in Applied Botany. Harvard University.
WELLS, EVELYN, Instructor in Biology, St. Mary's Seminary.
WELLS, L. J.. Graduate Student, University of Chicago.
WILDE, MARY H., Graduate Assistant. New Jersey College for Women.
WRIGHT, CHARLES I., Fellow in Physiology, University of Rochester Medical
School. YANCEY, PATRICK H., Graduate Student, St. Louis University.
Research Assistants
BECK, L. V., Teaching Fellow. New York University.
BERNSTEIN, ALAN. Research Assistant, New York University.
BUCK, LOUISE H., Research Assistant, New York University.
CAMPBELL, RAYMOND W.. Assistant. Fatigue Laboratory, Harvard University.
DAVIDSON, SYDNEY A., Williams College.
EISENBRANDT. W. II., Student, University of Maryland Medical School.
FRANCIS, DOROTHY S., Research Assistant, Memorial Hospital.
FRIEDMAN, HILDA, Assistant in Pathology. Washington University Medica! School.
FRIEDHEIM, ERNST A. H., Rockefeller Institute.
GRAND, CONSTANTINE G., Research Assistant, New York University.
GREENBERG, JACOB, Medical Student, Yale University Medical School.
HARRYMAN, ILENE, Research Assistant. Lilly Research Laboratory.
HEUSNF.K. A. P.. Student, Swarthmore College.
HOFFMAN, OLIVE D.. Research Assistant, New York University.
LAZAROVICH-HKEBELIANOVICH, MARA DE, Research Assistant, New York University.
MENDELSON, E. S.. Research Assistant, I'liivi-r^ity of Pennsylvania.
MENKIN, MIRIAM F.. Henry Phipps Institute.
OBERG, S. ALBERT, Harvard University.
PARKS, ELIZABETH K., Graduate Assistant in Zoology, Oberlin College.
RAVENSWAAY, VAN A. C.. Research Assistant, Washington University.
REYNOLDS, SARA JANE, Research Assistant, New York University.
RUDNICK, DOROTHEA. Research Assistant, University of Chicago.
SALOMON, KURT, Fellow of the Rockefeller Foundation. Rockefeller Institute for Medical Research.
SANDERS, ROSALTHA, Technician, Yale University.
SCHUBERT, MAXWELL, Assistant, Rockefeller Institute for Medical Research.
SCOTT, ALLAN C., Graduate Assistant. University of Pittsburgh.
SIIATTUCK, G. EDGAR. Assistant Instructor in Physiology, University of Pennsyl- vania.
SHEAR, M. J., Administrative Officer and Research Chemist, Pediatric Research Laboratory. Jewish Hospital.
SHLAER, SIMON, Research Assistant, Columbia University.
SMITH, M. DOREEN, Research Assistant, Memorial Hospital.
SVVANN, SHERLOCK, Research Associate. University of Illinois.
TOCKER, ALBERT M.. Student, Washington University Medical School.
WALLACE, EDITH M.. Artist and Research Assistant, Carnegie Institution of Wash- ington.
RKPORT OF THE DIRECTOR
Students BOTANY
BREED, HKLKN L., Student. Wellesley Collar.
BRUNEL, JULES, Assistant Professor of Botany, University of Montreal.
CHEEVER, CLARENCE A., Boston Society of Natural History.
DROUET, FRANCIS, Graduate Assistant, University of Missouri.
FORBES, JOHN M.. Student, Harvard University.
GLIDDEN, DOROTHY P.. Student, Smith College.
HOPKINS, MILTON, Student, Amherst College.
HUNTINGTON, EVELYN, Student, Vassar College.
KLOSE, THEODORE G., Instructor in Botany, Yassar College.
LOUGHRIDGE, GASPER A., Laboratory Assistant in Botany. Rutgers University.
McKEE. JEWEL C., University of Wisconsin.
OPPENHEIMER, JANE M., Student, Bryn Mawr College.
ROLAND, ALBERT E., Student, Acadia University.
SAFFORD, DEC i us W., Dartmouth College.
STEWART, PAUL A., Student, University of Rochester.
WILDE. MARY H.. Assistant, New Jersey College for Women.
EMBRYOLOGY
BAKER, E. G. STANLEY. Student, DePauw University.
BALLARD, OVERTON T.. University of Illinois.
BAMBER, LYLE E., Graduate Student and Assistant. University of Illinois.
CARTER, GEORGE H., Student, Amherst College.
DAWSON, HELEN L., Graduate Student, Washington University.
DERBYSHIRE, ARTHUR J., JR.. Harvard University.
EARL, RUTH R., Technician. College of the City of Xew York.
EATON, THEODORE H., JR.. Cornell University.
GREEN, DAVID E., Assistant, Washington Square College.
GUERLAC, HENRY E., Student. Cornell University.
HILEMAN. CLARA M., Instructor in Biology, Penn State College.
HJORTLAND, ARTHUR L.. Assistant, University of Illinois.
HUNNINEN, ARNE V., Student, Wesleyan University.
JOHNSON, ARLENE C., Student, Wheaton College.
JOHNSON, MYRA L., Student, Smith College.
LANE, MARY, Smith College.
LOEFER, JOHN B., Graduate Assistant in Biology, New York University.
MACKMULL, GULDEN, Demonstrator of Histology and Embryology, Baugh Insti- tute of Anatomy, Jefferson Medical College.
MAXWELL, FLORENCE J., Instructor, Carnegie Institute of Technology.
NICHOLS, ROWENA. Wellesley College.
PATCH, ESTHER M., Teacher of Biology, East Boston High School.
REID, MARION A., Instructor, Boston University Medical School.
RILEY, LENA C., Student, Wellesley College.
ROSENBAUM, LOUISE, Student, University of Pennsylvania.
SCOTT, JOHN P., University of Wyoming.
STANLEY. WILLARD F., Graduate Student and Research Assistant, University of Illinois.
WELLS, LEMEN J., Graduate Student, University of Chicago.
PHYSIOLOGY
APPELROT, SAMUEL, Fellow, Rockefeller Foundation. BEHNER, DOROTHY M., Assistant, New York University. CHANG, TSUNG H., Graduate Student, University of Chicag •.
26 MARINE BIOLOGICAL LABORATORY
CHEN, TCANG T.. Assistant in Biochemistry, Poking Union Medical College.
DANN, MARGARET, Assistant in Physiology, Cornell University Medical College.
DREW, WILLIAM B., Massachusetts Agricultural College.
DuBois, DELAFIELD, New York University.
FENG, TE-PEI, Graduate Student, University of Chicago.
GARDNER, EDITH McN., Assistant in Physiology, Vassar College.
GATES, FREDERICK L., Research Fellow, Harvard University.
HEGNAUER, ALBERT II., Fellow in Physiology, University of Rochester Medical
School.
LEITCH, JAMES L., University of California.
MONAGHAN, BETTY R., Assistant and Graduate Student, Washington University. OLIPHANT, JOSEPH F.. Instructor in Biology, Union College. OSTER, ROBERT H., Student, Williams College. PITTS, ROBERT F., Johns Hopkins University. SHAW, GRETCHEN, Graduate Student, University of Chicago. STEINER, MATTHEW M., Assistant in Biology, New York University. STEVENS, TIIELMA O., Graduate Assistant, Mt. Holyoke College. TANG, PEI-SUNG, Harvard University.
VACK, CHRISTINE M., Technician, Harvard University Medical School. VICARI, EMILIA M.. Research Associate, Cornell University Medical College. WOODWARD, ALVALYN E., Assistant Professor, University of Michigan.
PROTOZOOLOGY
BREHME, KATHERINE S., Barnard Colh
CARTER, HELEN D.. Elmira College.
COSTELLO, DoN.M.ii I'.. .v.iJ I Indrllu-rg. Detmit, Michigan.
EMBICH, JOHN R., Graduate Student, Columbia University.
FLAMMON, SISTER M. Mrun i., Sctmi Hill College.
FRYE, MARY ELIZABETH, Pennsylvania College fur Wnmen.
LHERISSON, CAMILLE. Professor of Biology, University of Haiti Medical School.
MASTEN. Lois I-'.., Khniru College.
MORGAN, \\'II.I.IK A.. Assistant Instructor in Biology, Coker College.
SCHOELT, ABRAHAM 11.. Graduate Student. Columbia University.
SCOTLAND, MINNIE B., Teacher, New York State College for Teachers.
SMITH, CLAIKI M., Hunter ColK-.ne.
STEINBERG, BERNHARD, Director of Laboratories and Research, Toledo Hospital.
WEISMAN, MAXWELL N., Fellow, College of the City of New York.
I \YKKTK!', RATE ZOOLOGY
ALDERMAN, EVANGELINE, Oberlin College.
BAUMGARTNKR, FREDERICK M.. Butler University.
BITTINGER, ISABEL, Radcliffe College.
BoARDMAN, EDWARD 'I'., Graduate Assistant, Johns Hopkins University.
BROWN, FRANK A., JR.. Harvard University.
CAMPBELL. DAX H., Student, Wahash College.
CARLSON, J. GORDON, Assistant in Zoology, University of Pennsylvania.
CHADWICK, CLAUDE S., Instructor in Biology, Vanderbilt University.
COHEN, ROSE S., Graduate Assistant, University of Cincinnati.
COLEMAN, LUCILLE, Agnes Scott College.
COPLAN, HELEN M., Student, Goucher College.
CRAIG. ROBERT L.. Student, Amherst College.
CROWELL. PRINCE S.. JR., Bowdoin College.
DEE, M. BARBARA, Assistant in Science, Jamaica Plain High School.
Di-:Roo. GRACE, Radcliffe College.
DERRICKSON. MARY B., Goucher College.
REPORT OF THE DIRECTOR
DORRIS, FRANCES S., Graduate Student, Yale University.
DOYLE, WILLIAM L., Johns Hopkins University.
EICHOLD, EVA C., Student, Newcomb College.
EVERETT, JOHN \V., Yale University.
FARBER, SEYMOUR M., University of Buffalo.
FENNELL, R. A., Graduate Student, Duke University.
FISHER, KENNETH C., Student and Assistant, Acadia University.
FRENCH, CHARLES S., Harvard University.
HAMBURGER, Louis P., JR., Johns Hopkins University.
HART, HELEN B., Student, Wellesley College.
HASTINGS, MARGARET, Student, Mt. Holyoke College.
HAYES, FREDERICK R., Assistant Professor of Zoology, Dalhousie University.
HEISS, ELIZABETH M., Assistant in Biology and Histology, Purdue University.
HEUSNER, ALBERT P., Swarthmore College.
HEWITT, DOROTHY C., Graduate Student, Yale University.
HOLLOWAY, MAY P., Teacher of Science, Burke School.
HUBBARD, RUTH A., Assistant, Cleveland Museum of Natural History.
JACKSON, JEANNETTE A., Graduate Assistant in Zoology, Syracuse University.
JOHNSON, DOROTHY F., Laboratory Assistant, Wellesley College.
KILLE, FRANK R., Assistant Professor of Biology, Birmingham-Southern College.
KROC, ROBERT L., Graduate Assistant, University of Wisconsin.
LEAVITT, BENJAMIN B., Instructor in Zoology, Dartmouth College.
MENZEL. ARTHUR E. O., National Tuberculosis Research Fellow, Presbyterian
Hospital.
MERRIMAN, DANIEL, Student, Harvard University. MORRIS, SAMUEL, Instructor in Zoology, University of Pennsylvania. PREST, MARGARET R., Graduate Assistant, Mt. Holyoke College. REDMOND, ALBERT C., Student, Hamilton College. REYNOLDS, ALBERT E., Assistant in Zoology, DePauw University. RITTER, M. ESTHER, Student, Wilson College. SHEA, MARGARET, Student, Oberlin College.
SIDEBOTHAM, RUTH S., Graduate Assistant in Zoology, Washington University. SNELL, PETER A., Fellow in Biology, Princeton University. SNOOK, THEODORE, Graduate Assistant in Zoology, Rutgers University. SWANSON, OSCAR E., Student, Antioch College. TIPTON, SAMUEL R., Graduate Student, Duke University.
TOWNSEND, GRACE, Instructor, Joliet Township High School and Junior College. TREAT, DOROTHY A., Assistant in Department of Education, Cleveland Museum of
Natural History.
WATERS, NELSON F., Graduate Student, Harvard University. WEED, MILTON R., Student, Wesleyan University. WOODRUFF, BETH H.. Graduate Assistant, Western Reserve University.
28
MARINE BIOLOGICAL LABORATORY
TABULAR VIEW OF ATTENi ) AXCE
1926 1927 1928 1929 1930
INVESTIGATORS— Total 252 294 323 329 337
Independent 156 209 217 234 217
Under Instruction 84 57 81 71 87
Research Assistants 12 28 25 24 33
STUDENTS— Total 141 141 133 125 136
Zoology 56 57 57 53 56
Protozoology 19 17 16 15 14
Embryology 28 32 29 28 27
Physiology 18 19 15 17 23
Botany 20 16 16 12 16
TOTAL ATTENDANCE 393 435 456 454 473
Less Persons registered as both students
and investigators 8 1 2 10 14
385 434 454 444 459
INSTITUTIONS HI-PHI si- NT-HI)— Total 119 111 111 123 126
By Investig-;it. .!> 84 8'» 80 96 95
By Students 60 (,3 66 64 71
SCHOOLS AND ACADEMIES RH.I'RESKNTED
My Investigators 1 1
My Students 4 4 1 1 4
r.iKF.icN INSTITUTIONS RKI-RKSKNTI.H
By Invest igator> 17 15 13 30 7
Bv Student- . 38832
4. SUBSCRIBING AXD COOPERATING INSTITUTIONS
Acadia University
. \mherst College
Antioch College
l'.ea\ er < 'ollege
Bowdoin College
Brown University
I'.rvn Mawr College
Rutler College
C. R. B. F.ducational Foundation
California Institute of Technology
Carnegie Institution of Washington
Chinese Educational Mission
Columbia University
Cornell University
Cornell University Medical College
Dalhousie University
Dartmouth College
I Vl'amv University
Duke University
F.lmira College
General Education Board
Goucher College
Hamilton College
I harvard University
Harvard University Medical School
Howard University
I lunter College
Indiana University
Industrial & Engineering Chemistry.
of the American Chemical Society Johns Hopkins University Johns Hopkins University Medical
Scliool
F.li Lilly & Co.
Memorial Hospital of X. Y. City rUorehouse College Mount TTolyoke College National Research Council New York State Department of
Health
New York University Oherlin College
REPORT OF THE DIRECTOR
29
Pennsylvania College for Women Princeton University Radcliffe College Rockefeller Foundation Rockefeller Institute for Medical
Research
Rutgers University Seton Hill College Smith College Sophie Newcomb College Southwestern St. Louis University Swarthmore College Tufts College Union College United States Dept. of University of Buffalo University of Chicago University of Cincinnati University of Illinois University of Michigan University of Missouri University of Nebraska Universitv of Pennsylvania
Agriculture
University of Pennsylvania Medical
School
University of Pittsburgh University of Rochester University of Tennessee University of Virginia University of Wisconsin Vanderbilt University Medica
School
Yassar College Wabash College Washington University Washington University Medical
School
AVellesley College Wesleyan University Western Reserve University West Virginia State College West Virginia University Wheaton College Wistar Institute of Anatomy and
Biology Yale Universitv
SCHOLARSHIP TABLES
Lucretia Crocker Scholarships for Teachers in Boston.
Scholarship of $100 supported by a friend of the Laboratory since 1898.
The Edwin S. Linton Memorial Scholarship of Washington and Jefferson
College.
The Bio Club Scholarship of the College of the City of New York. Ida H. Hyde Scholarship of the University of Kansas.
5. EVENING LECTURES, 1930 Tuesday, July 1
DR. W. B. SCOTT " Xew Light on the Development and
Migrations of American Mam- mal.-."
"Prospects and Problems of Ocean- ography."
Tuesday, July 8
DR. HENRY B. BIGELOW
Tuesday, July 15
DR. H. S. IENNINGS .
"Heredity and Mutation in Relation
to Environment in Protozoa." Friday, July 18
DR. EDWARD MELLAXBY " Food Deficiencies."
Tuesday. July 22
DR. F. R. LILLIE " The Action of the Sex Hormones in
the Fowl : An Account of the Chi- cago Experiments."
30 MARINE BIOLOGICAL LABORATORY
Tuesday, July 29
DR. LEONOR MICHAELIS " The Reversible Oxidizable-reducible
Systems Found in Living Organ- isms." Tuesday, August 5
DR. E. F. DuBois " Recent Progress in the Field of Re- spiratory Metabolism." Tuesday, August 12
DR. E. CHATTON "A Study of the Dinorlagellate, Poly-
krikos Schwartzi as a Basis for the Discussion of Some Problems of General Cytology." Tuesday, August 19
Tin-: REYNOLD A. SPAETH Mi M- RIAL LECTURE, delivered by DR.
HARDOLPH WASTEXEYS " Protein Synthesis."
Thursday, August 21
DR. II. H. GRAN " The Productivity of the Ocean."
Tuesday, August 26
DR. D. \Y. BKOXK " Xerve Impulse Rhythms and the
Control of Movement." Tuesday, September 2
DR. G. C. HIRSCII " The Problem of Restitution with
Special Regard to the Phenomena of Secretion." Tuesday, September 9
DR. G. E. HUTCHINSON "The Hydrobiology of Arid and
Semi-arid Regions."
SPECIAL LECTURES AND MOTION PICTURES
Monday, August 1 1
" The lllyria Expedition to the Galapagos, the South Sea Islands, the Xew Hebrides, the Solomon Islands, Xew Guinea, Bali and Angkor." Exhibited by MR. CORNII irs CRANE, the Leader of the Expedition.
Thursday, August 14
" The Florida Everglades and the Proposed Tropic Everglades National Park." Illustrated with colored lantern slides. MR. ERNEST F. COE, Chairman of the Tropic Everglades Park Association.
Thursday, August 28 Motion pictures.
"William Harvey and the Circulation of the Blood." Arranged by SIR THOMAS LEWIS.
"The Early Development of the Rabbit Egg." Du. \Y. II. LEWIS and DR. I'. W. GREGORY.
"The Life Cycle of the Oyster." Prepared by the AMERICAN MUSEUM OK NATURAL HISTORY.
Friday, August 29
" Motion Pictures of Marine and Fresh Water Protozoa of the Woods Hole Region," RTTH B. I IOWI.AXD, Department of Biology, Xew York University.
REPORT OF THE DIRECTOR
6. SHORTER SCIENTIFIC PAPERS, 1930
Tuesday, June 24
DR. BALDUIN LUCKE AND
DR. MORTON McCuTCHEON "The Effect of Injury on Cellular
Permeability to Water."
DR. M. M. BROOKS "The Relation between rH and the
Penetration of Oxidation-reduction Indicators."
DR. S. C. BROOKS " Accumulation of Ions in Living
Cells."
DR. M. H. JACOBS, iMR. A. K. PAR- PART, DR. W. A. SMITH AND MR.
G. E. SHATTUCK " The Permeability of the Erythrocyte
to Urea." Friday, June 27
DR. B. H. WILLIER " The Developmental Relations of the
Heart and the Liver in Chorio- allantoic Grafts."
DR. E. E. JUST " Cortical Protoplasm and Vital Phe- nomena."
DR. WILLIAM C. YOUNG " The Post-testicular History of Sper- matozoa and Reproduction in the Male Guinea Pig."
DR. L. V. HEILBRUNN " The Action of Ultra-violet Rays on
Arbacia Egg Protoplasm." Thursday, July 3
DR. WALTER S. ROOT AND
DR. CHARLOTTE HAYWOOD " The Effect of Carbon Dioxide upon
the Rate of Oxygen Consumption and of Cleavage of the Arbacia Egg."
DR. L. MlCHAELIS AND
DR. K. SALOMON " Respiration of Ervthrocytes."
DR. R. W. GERARD " Observations on the Metabolism of
the Coccus, Sarcina lutea."
DR. E. S. G. BARRON " The Effect of Methylene Blue upon
the Respiration of Normal and of Cancer Tissue." Friday, July 11
DR. W. R. TAYLOR " Chromosome Structure in Meiosis of
Gasteria."
DR. B. M. DUGGAR " New Technique and Some Adsorp- tion Studies with Virus Diseases of Plants." Thursday, July 17
DR. VERA KOEHRIXG "Some Cytological Relationships in
Narcosi-."
DR. ELEANOR H. SLIFER " The Mitotic Activity in the Devel- oping Grasshopper Egg."
MARINE BIOLOGICAL LABORATORY
DR. A. F. HUETTXER " Spermatogenesis in Drosophila mel-
anogaster."
DR. HOPE HiBBARn " Cytological Studies on the Silk Gland
of Bombyx mori." Friday, July 25
DR. S. GELFAX " The All-or-Xone Law in Muscle."
DR. \Y. R. AMBERSOX. MR. A. K.
PARPART AXD Miss GERTRUDE SAN- DERS " Low Voltage Elements of the Action
Potential \\'ave of Nerve."
DR. F. O. SCHMITT " The Effect of Cyanides and Carbon
Monoxide on Nerve." Saturday, July 26
DR. M. Driirissox "Cardiac Automatism in Inverte- brates."
DR. \V. F. GAKREV " Observations on the 1 leart of Limu-
lus."
DR. D. J. EDWARDS " The Action of Pressure on Some
1 'hysiological Processes."
DR. G. H. BISHOP " The Influence of lodo-acetic Acid on
Muscle Contracture."
DR. F. H. PRATT " Experiments on the Terminal Nerve- muscle Unit."
DR. I). \V. BROXK " Graded Mu>cular Contractions."
DR. K. \\'. GERARD " Nerve Metabolism and Asphyxia."
DR. R. S. LII.I.IE " Recovery in the Passive Iron Wire
Model.""
DR. A. M. MOXXIKR " Mathematical Analysis Applied to
the Functions of the Nervous Sys- tem."
DR. 1 1. DAVIS " Re-education and Modification of
Reflexes."
DR. G. P. McCorrii " Patterns of Some Extra-ocular Re-
tle\e> in the Cat."
DR. P. I! \K-i) " The Piehavior of a Cat without the
Telencephalon." Friday, Au.uu-t 1
DR. PAUL S. llr.\sir.\w "Some Biological Effects Produced
by Alpha Particles on Drosophila Eggs."
MR. WARE CATTEI.I " The Effect of X-Rays upon the
Growth of the Wheat Seedling."
DR. W. C. CURTIS " Effects of X-Rays upon Regenera- tion."
DR. CHARLES PACKARD "The Relation between Division Rate
and Susceptibility to Radiation." Friday, August 8
DR. HELEN M. MILLER "Life Cycle of a Bisexual Rotifer."
DR. TRACY M. SONNEBORX "Cause, Inheritance, and Effects of
the Chain-forming Tendency in the Ciliate Protozoan, Colpidium."
REPORT OF THE DIRECTOR
DR. RUTH STOCKING LYNCH " The Effects of Long-continued
Starvation in a Rotifer in Relation to Inheritance."
DR. H. W. STUNKARD " The Life History of Cryptocotyle
lingua."
Friday, August 15
DR. MARY S. M \cDoucALL "A Mutation in Chilodon uncinatus
Produced by Ultra-violet Radiation —A Preliminary Report."
DR. R. D. MANWELL " The Effect of Quinine and Plas-
moquin on Avian Malaria."
DR. E. CHATTON " The Asymmetric Motile Stages of
Epistylis and the Question of the So-called Longitudinal Division of the Vorticellidae."
DR. RUTH B. ROWLAND " Cine-photomicrograph of Microin-
jection of Vacuolated Protoplasm."
Friday, August 22
DR. CALVIN B. BRIDGES " The Neutralization of the Effects of
Deficiencies through Duplications of the Same Chromosome Material."
DR. A. H. STURTEVANT "A Peculiar Sex-ratio in Drosophila
obscura."
DR. HELEN REDFIELD " Studies of Crossing-over in Droso- phila."
DR. JACK SCHULTZ " The Eye of Pigments of Drosophila."
Friday, August 29
DR. H. B. GOODRICH AND
MR. I. B. HANSEN "Embryonic Development of Men- del ian Characters in the Goldfish."
DR. H. A. ABRAMSON " The Isoelectric Point of Mammalian
Red Blood Cells."
DR. E. N. HARVEY AND
MR. A. L. LOOMIS " The Microscope-centrifuge."
Friday, September 5
DR. J. M. JOHLIN " The Extraction of Micro-organisms."
DR. W. E. GARREY AND
DR. W. R. BRYAN " Alkalosis in Relation to Tetany fol- lowing High Temperatures after Parathyroidectomy."
DR. K. BLANCHARD " Catalysis of Condensation Reactions
by Amino-acids."
DR. L. MlCHAELIS AND
DR. M. SCHUBERT " Metal Complex Compounds of Thio-
glycollic Acid," 3
34 MARINE BIOLOGICAL LABORATORY
7. MEMBERS OF THE CORPORATION
1. LIFE MEMBERS
ALLIS, MR. E. P., JR., Palais Carnoles, Menton, France.
ANDRF.WS, MRS. GWENDOLEN FOULKE, Baltimore, Md.
BILLINGS, MR. R. C., 66 Franklin St., Boston, Mass.
CONKLIN, PROF. EDWIN G., Princeton University, Princeton, N. J.
COOLIDGE, MR. C. A., Ames Building, Boston, Mass.
CRANE, MR. C. R., New York City.
EVANS, MRS. GLENDOWER, 12 Otis Place, Boston, Mass.
FAY, Miss S. B., 88 Mt. Vernon St., Boston, Mass.
FOOT, Miss KATHERIXK, Care of Morgan Harjes Cie, Paris, France.
GARDINER, MRS. E. G., Woods Hole, Mass.
JACKSON, Miss M. C., 88 Marlboro St., Boston, Mass.
JACKSON, MR. CHAS. C., 24 Congress St., Boston, Mass.
KIDDER, MR. NATHANIEL T., Milton, Mass.
KING, MR. CHAS. A.
LEE, MRS. FREDERIC S., 279 Madison Are., New York City, N. Y.
LOWELL, MR. A. LAWRENCE, 17 Quincy St., Cambridge, Mass.
MEANS, DR. JAMES HOWARD, 15 Chestnut St., Boston. Mass.
MERRIMAN, MRS. DANIEL, 73 Bay State Road, Boston, Mass.
MINNS, Miss SUSAN, 14 Louisburg Square, Boston, Mass.
MORGAN, MR. J. PIERPONT, JR., Wall and Broad Sts., New York City,
N. Y. MORGAN, PROF. T. H., Director of Biological Laboratory, California
Institute of Technology, Pasadena, Calif. MORGAN, MRS. T. H., Pasadena, Calif. NOYES, Miss EVA J. OSBORN, PROF. HENRY F., American Museum of Natural History. New
York, N. Y.
PHILLIPS, MRS. JOHN C., Windy Knob, Wenham, Mass. PORTER, DR. H. C., University of Pennsylvania, Philadelphia, Pa. SEARS, DR. HKXKY F., Xf> Bracon St., Boston, Mass. SHEDD, MR. E. A. THORNDIKE, DR. EDWARD L., Teachers College, Columbia University,
New York City, N. Y.
TRELEASE, PROF. WILLIAM, University of Illinois, Urbana, 111. WARE, Miss MARY L., 41 Brimmer St., Boston, Mass. WILLIAMS, MRS. ANNA P., 505 Beacon St., Boston, Mass. WILSON, DR. E. B., Columbia University, New York City, N. Y.
REPORT OF THE DIRECTOR
2. REGULAR MEMBERS, AUGUST, 1930
ADAMS, DR. A. ELIZABETH, Mount Holyoke College, South Hadley,
Mass. ADDISON, DR. W. H. F., University of Pennsylvania Medical School,
Philadelphia, Pa. ADOLPH, DR. EDWARD F., University of Rochester, School of Medicine
and Dentistry, Rochester, N. Y.
ALLEE, DR. W. C., University of Chicago, Chicago, 111. ALLEN, PROF. CHARLES E., University of Wisconsin, Madison, Wis. ALLEN, PROF. EZRA, New York Homeopathic Medical College, New
York City, N. Y.
ALLYN, DR. HARRIET M., Mount Holyoke College, South Hadley, Mass. AMBERSON, DR. WILLIAM R., University of Tennessee, Memphis, Tenn. ANDERSON, DR. E. G., California Institute of Technology, Pasadena,
Calif.
AUSTIN, DR. MARY L., Wellesley College, Wellesley, Mass. BAITSELL, DR. GEORGE A., Yale University, New Haven, Conn. BAKER, DR. E. H., 5312 Hyde Park Boulevard, Hyde Park Station,
Chicago, 111. BALDWIN, DR. F. M., University of Southern California, Los Angeles,
Calif.
BECKWITH, DR. CORA J., Vassar College, Poughkeepsie, N. Y. BEHRE, DR. ELINOR H., Louisiana State University, Baton Rouge, La. BENNITT, DR. RUDOLF, University of Missouri, Columbia, Mo. BIGELOW, PROF. R. P., Massachusetts Institute of Technology, Cam- bridge, Mass.
BINFORD, PROF. RAYMOND, Guilford College, Guilford College, N. C. BISSONNETTE, DR. T. H., Trinity College, Hartford, Conn. BLANCHARD, PROF. KENNETH C., New York University, Washington
Square College, New York City, N. Y. BODINE, DR. J. H., University of Iowa, Iowa City, la. BORING, DR. ALICE M., Yenching University, Peking, China. BOWLING, Miss RACHEL, Columbia University, New York City, N. Y. Box, Miss CORA M., University of Cincinnati, Cincinnati, O. BRADLEY, PROF. HAROLD C., University of Wisconsin, Madison, Wis. BRAILEY, Miss MIRIAM E., 800 Broadway, Baltimore, Md. BRIDGES, DR. CALVIN B., California Institute of Technology, Pasadena,
Calif.
BRONK, DR. D. W., University of Pennsylvania, Philadelphia. Pa. BROOKS, DR. S. C., University of California, Berkeley, Calif. BUCKINGHAM, Miss EDITH N., Sudbury, Mass.
36 MARINE BIOLOGICAL LABORATORY
BUDINGTON, PROF. R. A., Oberlin College, Oberlin, O. BULLINGTON, DR. W. E., Kaiiclolph-Macon College, Ashland, Ya. BUMPUS, PROF. H. C., 76 Carlton Road. \\'al)an, Mass. BYRNES, DR. ESTHER F., 1803 North Camac Street, Philadelphia. Pa. CALKINS, PROF. GARY X., Columbia University, New York City, N. Y. CALVERT, PROF. PHILIP P., University of Pennsylvania, Philadelphia,
Pa.
CARLSON, PROF. A. J., University of Chicago, Chicago, 111. CAROTHERS, DR. ELEANOR E., University of Pennsylvania, Philadelphia.
Pa. CARROLL, PROF. MITCHEL, Franklin and Marshall College, Lancaster,
Pa.
CARVER, PROF. GAIL L., Mercer University, Macon, Ga. CATTELL, DR. McKEEN, Cornell University Medical College, New
York City, N. Y.
CATTELL, PROF. J. McKEEN, Garrison-on-Hudson, N. Y. CATTELL, MR. WARE, Garrison-on-Hudson, N. Y. CHAMBERS, DR. ROBERT, Washington Square College, New York Uni- versity, Washington Square, New York City, N. Y. CHARLTON. DR. HARRY H., University of Missouri Columbia, Mo. CHATTON, DR. EDOUARD. University of Strasbourg, Strasbourg, France. CIIIDESTER, PROF. F. E., West Virginia University, Morgantown,
W. Va
CHILD, PROF. C. M., University of Chicago, Chicago, 111. CLAPP, PROF. CORNELIA M., Montague, Mass.
CLARK, PROF. E. R., University of Pennsylvania, Philadelphia, Pa. CLELAND, PROF. RALPH E., Gouchcr College, Baltimore, Md. CLOWES, PROF. G. H. A., Eli Lilly & Co., Indianapolis, Ind. COE, PROF. W. R., Yale University, New Haven, Conn. COHN, DR. EDWIN J., 183 Brattle St., Cambridge, Mass. COLE, DR. I ALBERT C., Williams College, Williamstown, Mass. COLE, DR. LEON J., College of Agriculture, Madison. \YU. COLLETT, DR. MARY E., Western Reserve University, Cleveland, O. COLLEY, MRS. MARY W., 36 Argyle Place, Rockville Centre, Long
Island, N. Y.
COLTON, PROF. II. S., Box 127, Flagstaff, Ariz. CONNOLLY, DR. C. J., Catholic University, Washington, D. C. COPELAND, PROF. MANTON, Bowdoin College, Brunswick. Me. COWDRY, DR. E. V., Washington University, St. Louis, Mo. CRAMPTON, PROF. II. E., Barnard College, Columbia University, New
York City, N. Y. CRANE, MRS. C. R., Woods Hole, Mass.
RFPORT OF THE DIRECTOR
CURTIS, DR. MAYNIK R., Crocker Laboratory, Columbia University,
New York City, N. Y.
CURTIS. PROF. W. C., University of Missouri, Columbia, Mo. DAVIS, DR. ALICE R., Castle Point. Hoboken, N. J. DAVIS, DR. DONALD W., College of William and Mary, Williamsburg,
Va.
DAWSON, DR. A. B., Harvard University, Cambridge, Mass. DAWSON, DR. J. A., The College of the City of New York, New York
City, N. Y.
DEDERER, DR. PAULINE H., Connecticut College, New London, Conn. DELLINGER, DR. S. C., University of Arkansas, Fayetteville, Ark. DODDS, PROF. G. S., Medical School, University of West Virginia, Mor-
gantown, W. Va.
DOLLEY, PROF. WILLIAM L., University of Buffalo, Buffalo, N. Y. DONALDSON, PROF. H. H., Wistar Institute of Anatomy and Biology,
Philadelphia, Pa.
DONALDSON, DR. JOHN C., University of Pittsburgh, School of Med- icine, Pittsburgh, Pa.
DREW, PROF. OILMAN A., Eagle Lake, Florida. Du Bois, DR. EUGENE F., Cornell University Medical College, New
York City, N. Y.
DUGGAR, DR. BENJAMIN M., University of Wisconsin, Madison, Wis. DUNGAY, DR. NEIL S., Carleton College, Northfield, Minn. DUNN, DR. L. C., Columbia University, New York City, N. Y. EDWARDS, DR. D. J., Cornell University Medical College, New York
City, N. Y.
ELLIS, DR. F. W., Monson, Massachusetts. FARNUM, DR. LOUISE W., Hsiang-Ya Hospital, Changsha, Hunan,
China.
FAURE-FREMIET, PROF. EMMANUEL, College de France, Paris, France. FENN, DR. W. O., Rochester University, School of Medicine, Rochester,
N. Y.
FIELD, Miss HAZEL E., Occidental College, Los Angeles, Calif. FORBES, DR. ALEXANDER, Harvard University Medical School, Boston,
Mass.
FRY, DR. HENRY J., Washington Square College, New York City, N. Y. GAGE, PROF. S. H., Cornell University, Ithaca, New York. GARREY, PROF. W. E., Vanderbilt University Medical School, Nashville.
Term.
GATES, DR. F. L., 31 Fayerweather St., Cambridge, Mass. GATES, PROF. R. RUGGLES, University of London, London, England. GEISER, DR. S. W., Southern Methodist University, Dallas, Tex.
38 MARINE BIOLOGICAL LABORATORY
GLASER, PROF. O. C, Amherst College, Amherst, Mass.
GLASER, PROF. R. W., Rockefeller Institute for Medical Research,
Princeton, N. J. GOLDFORB, PROF. A. J., College of the City of Xew York, Xcw York
City, N. Y.
GOODRICH, PROF. H. B., Wesleyan University, Middletown, Conn. GRAHAM, DR. J. Y., University of Alabama, University, Ala. GRAVE, PROF. B. H., DePauw University, Greencastle, Ind. GRAVE, PROF. CASWELL, Washington University, St. Louis, Mo. GRAY, PROF. IRVING E., Duke University, Durham, X. C. GREENMAN, PROF. M. J., Wistar Institute of Anatomy and Biology,
Philadelphia, Pa. GREGORY, DR. LOUISE H., Barnard College, Columbia University, New
York City, N. Y.
GUTHRIE, DR. MARY J., University of Missouri, Columbia, Mo. GUYER, PROF. M. F., University of Wisconsin, Madison, Wis. HAGUE, DR. FLORENCE, Sweet Briar College, Sweet Briar, Va. HALL, PROF. FRANK G., Duke University, Durham, X1. C. HANCE, DR. ROBERT T., University of Pittsburgh, Pittsburgh,. Pa. HARGITT, PROF. GEORGE T., Duke University, Durham. X. C. HARMAN, DR. MARY T., Kansas State Agricultural College, Manhattan,
Kans.
HARPER, PROF. R. A., Columbia University, New York City, N. Y. HARRISON, PROF. Ross G., Yale University, New Haven, Conn. HARVEY, MRS. E. N., Princeton, N. J.
HARVEY, PROF. E. N., Princeton University, Princeton, N. J. HAYDEN, DR. MARGARET A., Wellesley College, Wellesley, Mass. HAYWOOD, DR. CHARLOTTE, Mount llolyoke College, South Hadley,
Mass. HAZEN, DR. T. E., Barnard College, Columbia University, New York
City, N. Y.
HEATH, PROF. HAROLD, Pacific Grove, California. HECHT, DR. SELIG, Columbia University, New York City, XT. Y. HEGNER, PROF. R. W., Johns Hopkins University, Baltimore, Md. HEILBRUNN, DR. L. V., University of Pennsylvania. Philadelphia, Pa. HESS, PROF. WALTER N., Hamilton College, Clinton, N. Y. HINRICHS, DR. MARIE A., University of Chicago, Chicago, 111. HISAW, DR. F. L., University of Wisconsin, Madison, Wis. HOADLEY, DR. LEIGH, Harvard University, Cambridge, Mass. HOGUE, DR. MARY J., 503 N. High St., West Chester, Pa. HOLMES, PROF. S. J., University of California. Berkeley, Calif. HOOKER, PROF. DAVENPORT, University of Pittsburgh, Pittsburgh, Pa.
REPORT OF THE DIRECTOR
39
HOPKINS, DR. HOYT S., New York University, College of Dentistry,
New York City, N. Y.
HOWARD, DR. HARVEY J., Washington University, St. Louis, Mo. HOWE, DR. H. E., 2702 36th St., N. W., Washington, D. C. HOYT, DR. WILLIAM D., Washington and Lee University, Lexington,
Va. HUMPHREY, MR. R. R., University of Buffalo, School of Medicine,
Buffalo, N. Y.
HYMAN, DR. LIBBIE H., University of Chicago, Chicago, 111. INMAN, PROF. ONDESS L., Antioch College, Yellow Springs, O. IRWIN, DR. MARIAN, Rockefeller Institute, New York City, N. Y. JACKSON, PROF. C. M., University of Minnesota, Minneapolis, Minn. JACOBS, PROF. MERKEL H., University of Pennsylvania, Philadelphia,
Pa.
JENNINGS, PROF. H. S., Johns Hopkins University, Baltimore, Md. JEWETT, PROF. J. R., Harvard University, Cambridge, Mass. JOHNSON, PROF. GEORGE E., State Agricultural College, Manhattan,
Kans.
JONES, PROF. LYNDS, Oberlin College, Oberlin, O. JUST, PROF. E. E., Howard University, Washington, D. C. KEEFE, REV. ANSELM M., St. Norbert College, West Depere, Wis. KENNEDY, DR. HARRIS, Readville, Mass.
KINDRED, DR. J. E., University of Virginia, Charlottesville, Va. KING, DR. HELEN D., Wistar Institute of Anatomy and Biology, Phila- delphia, Pa.
KING, DR. ROBERT L., State University of Iowa, Iowa City, la. KINGSBURY, PROF. B. F., Cornell University, Ithaca, N. Y. KIRKHAM, DR. W. B., Springfield College, Springfield. Mass. KNAPKE, REV. BEDE, St. Bernard's College, St. Bernard, Ala. KNOWER, PROF. H. McE., Albany Medical College, Albany, N. Y. KNOWLTON, PROF. F. P., Syracuse University, Syracuse, N. Y. KOSTIR, DR. W. J., Ohio State University, Columbus, O. KRIBS, DR. HERBERT, 202A Copley Road, Upper Darby, Pa. KUYK, DR. MARGARET P., Westbrook Ave., Richmond, Va. LANCEFIELD, DR. D. E., Columbia University, New York City, N. Y. LANGE, DR. MATHILDE M., Wheaton College, Norton, Mass. LEE, PROF. F. S., College of Physicians and Surgeons, New York City,
N. Y.
LEWIS, PROF. I. F., University of Virginia, Charlottesville, Va. LEWIS, PROF. W. H., Johns Hopkins University, Baltimore, Md. LILLIE, PROF. FRANK R., University of Chicago, Chicago, 111. LILLIE, PROF. RALPH S., University of Chicago, Chicago, 111.
40 MARINE BIOLOGICAL LABORATORY
LINTON, PROF. EDWIN, University of Pennsylvania, Philadelphia, Pa. LOEB, PROF. LEO, Washington University Medical School, St. Louis,
.Mo.
LOEB, MRS. LEO, 812 Boland Place, St. Louis, Mo. LOWTHER, MRS. FLORENCE DeL., Barnard College, Columbia University,
New York City, N. Y.
LUCKE, PROF. BALDUIN, University of Pennsylvania, Philadelphia. Pa. LUND, DR. E. J., University of Texas, Austin, Tex. LUSCOMBE, MR. W. O., Woods Hole, Mass.
LYNCH, DR. CLARA J., Rockefeller Institute, New York City, N. Y. LYNCH, DR. RUTH STOCKING, Johns Hopkins University, Baltimore,
Md.
LYON, PROF. E. P., University of Minnesota, Minneapolis, Minn. MACDOUGALL, DR. MARY S., Agnes Scott College, Decatur, Ga. McCLUNG, PROF. C. E., University of Pennsylvania, Philadelphia, Pa. McGEE, DR. ANITA NEWCOMB, Box 363, Southern Pines, N. C. MCGREGOR, DR. J. H., Columbia University, New York City, N. Y. McMuRRiCH, PROF. J. P., University of Toronto, Toronto, Canada. McNAiR, DR. G. T., 1624 Alabama St., Lawrence, Kans. MACKLIN, DR. CHARLES C., School of Medicine, University of Western
Ontario, London, Canada.
MALONE, PROF. E. F., University of Cincinnati, Cincinnati, O. MANWELL, DR. REGINALD D., School of Hygiene and Public Health,
Johns Hopkins University, Baltimore, Md. MARTIN, PROF. E. A., College of the City of New York, New York
City, N. Y.
MAST, PROF. S. O., Johns Hopkins University, Baltimore, Md. MATHEWS, PROF. A. P., University of Cincinnati, Cincinnati, O. MAYOR, PROF. JAM is \V., Union College, Schenectady, N. Y. MEDES, DR. GRACE, University of Minnesota, Minneapolis, Minn. MEIGS, DR. E. B., Dairy Division Experiment Station, Beltsville. Md. MEIGS, MRS. E. B., 1736 M St., N. W., Washington, D. C. METCALF, PKOF. M. M., Johns Hopkins University, Baltimore, Md. METZ, PROF. CHARLES W., Carnegie Institution of Washington, Cold
Spring Harbor, Long Island, N. Y.
MICHAELIS, DR. LEONOR, Rockefeller Institute, New York City, N. Y. MILLER, DR. HELEN M., Johns Hopkins University, Baltimore. Md. MINER, DR. ROY W., American Museum of Natural History, New
York City, N. Y.
MITCHELL, DK. PHILIP II., Brown University, Providence. R. I. MOORE, DR. CARL R., University of Chicago, Chicago, 111. MOORE, PROF. GEORGE T., Missouri Botanical Garden. St. Louis, Mo.
REPORT OF THE DIRECTOR 41
MOORE, PROF. J. PI-:RCY, University of Pennsylvania, Philadelphia, Pa. MORGULIS, DR. SERGIUS, University of Nebraska, Lincoln, Nebr. MORRILL, PROF. A. D., Hamilton College, Clinton, N. Y. MORRILL, PROF. C. V., Cornell University Medical College, New York
City, N. Y.
MULLER, DR. H. J., University of Texas, Austin, Tex. NABOURS, DR. R. K., Kansas State Agricultural College, Manhattan,
Kans.
NEAL, PROF. H. V., Tufts College, Tufts College, Mass. NEWMAN, PROF. H. H., University of Chicago, Chicago, 111. NICHOLS, DR. M. LOUISE, Dreycott Apartments, Haverford, Pa. NOBLE, DR. GLADWYN K., American Museum of Natural History, New
York City, N. Y. NONIDEZ, DR. JOSE F., Cornell University Medical College, New York
City, N. Y.
OKKELBERG, DR. PETER, University of Michigan, Ann Arbor, Mich. OSBURN, PROF. R. C., Ohio State University', Columbus, O. OSTERHOUT, PROF. \Y. ]. V., Rockefeller Institute, New York City,
N. Y.
PACKARD, DR. CHARLES, Columbia University, Institute of Cancer Re- search, 1145 Amsterdam Ave., New York City, N. Y. PAGE, DR. IRVINE H., Presbyterian Hospital, New York City, N. Y. PAPANICOLAOU, DR. GEORGE N., Cornell University Medical College,
New York City, N. Y. PAPPENHEIMER, DR. A. M., Columbia University, New York City,
N. Y.
PARKER, PROF. G. H., Harvard University, Cambridge, Mass. PATON, PROF. STEWART, Princeton University, Princeton, N. J. PATTEN, DR. BRADLEY M., Western Reserve University, Cleveland, O. PATTEN, PROF. WILLIAM, Dartmouth College, Hanover, N. H. PAYNE, PROF. F., University of Indiana, Bloomington, Ind. PEARL, PROF. RAYMOND, Institute for Biological Research, 1901 East
Madison Street, Baltimore, Md.
PEARSE, PROF. A. S., Duke University, Durham, N. C. PEEBLES, PROF. FLORENCE, California Christian College, Los Angeles,
Calif.
PHILLIPS, DR. E. F., Cornell University, Ithaca, N. Y. PHILLIPS, DR. RUTH L., Western College, Oxford, O. PIKE, PROF. FRANK H., 437 West 59th St., New York City, N. Y. PINNEY, DR. MARY E., Milwaukee-Downer College, Milwaukee, Wis. PLOUGH, PROF. HAROLD H., Amherst College, Amherst, Mass. POLLISTER, DR. A. W., Columbia University, New York City, N. Y.
42 MARINE BIOLOGICAL LABORATORY
POND, DR. SAMUEL E., University of Pennsylvania. School of Medicine,
Philadelphia, Pa.
PRATT, DR. FREDERICK H., Boston University, School of Medicine, Bos- ton, Mass.
RAFFEL, DR. DANIEL, Johns Hopkins University, Baltimore. Md. RAND, DR. HERBERT W., Harvard University, Cambridge, Mass. RANKIN, PROF. W. M.. Princeton University, Princeton, N. J. REDFIELD, DR. ALFRED C., Harvard University Medical School, Boston,
Mass. REESE, PROF. ALBERT M., West Virginia University, Morgantown,
W. Va.
REINKE, DR. E. E., Vanderbilt University, Nashville, Tenn. REZNIKOFF, DR. PAUL, Cornell University Medical College, New York
City, N. Y.
RHODES, PROF. ROBERT C., Emory University, Atlanta, Ga. RICE, PROF. EDWARD L., Ohio Wesleyan University, Delaware, O. RICHARDS, PROF. A., Unm-rsity of Oklahoma, Norman, Oklahoma. RIGGS, MR. LAWRASON, JR., 25 Broad St., New York City, N. Y. ROBERTSON, PROF. W. R. B., 1803 Anderson Street, Manhattan, Kan. ROGERS, PROF. CHARLES G., Oberlin College, Oberlin, O. ROMER, DR. ALFRED S., University of Chicago, Chicago, 111. ROOT, DR. W. S.. Syracuse Medical School, Syracuse, N. Y. SAMPSON, DR. MYRA M., Smith College, Northampton, Mass. SANDS, Miss ADELAIDE G., 562 King St., Port Chester, N. Y. SCHRADER, DR. FRANZ, Department of Zoology, Columbia University,
New York City, N. Y.
SCHRAMM, PROF. J. R., University of Pennsylvania, Philadelphia, Pa. SCOTT, DR. ERNEST L., Columbia University, New York City, N. Y. SCOTT, PROF. G. G., College of the City of New York, New York City,
N. Y.
SCOTT, PROF. JOHN W., University of Wyoming, Laramie, Wyoming. SCOTT, PROF. WILLIAM B., 7 Cleveland Lane, Princeton, N. J. SHULL, PROF. A. FRANKLIN, University of Michigan, Ann Arbor, Mich. S HUM WAY, DR. WALDO, University of Illinois, Urbana, 111. SIVICKIS, DR. P. B., Pasto deze 130, Kaunas, Lithuania. SNOW, DR. LAETITIA M., Wellesley College, Wellesley, Mass. SNYDF.R, PROF. CHARLES D., Johns Hopkins University Medical School,
Baltimore, Md.
SOLLMAN, DR. TORALD, Western Reserve University, Cleveland, O. SONNEBORN, DR. T. M., Johns Hopkins University, Baltimore, Md. SPEIDEL, DR. CARL C., University of Virginia, University, Va. SPENCER, PROF. H. J., 24 West 10th St., New York City, N. Y.
REPORT OF THE DIRECTOR 43
STARK, DR. MARY B., New York Homeopathic Medical College and Flower Hospital, New York City, N. Y.
STOCKARD, PROF. C. R., Cornell University Medical College, New York City, N. Y.
STOREY, DR. ALMA G., Mount Holyoke College, South Hadley, Mass.
STRONG, PROF. O. S., College of Physicians and Surgeons, 630 West 168th Street, New York City, N. Y.
STUNKARD, DR. HORACE W., New York University, University Heights, N. Y.
STURTEVANT, DR. ALFRED H., California Institute of Technology, Pas- adena, Calif.
SUM WALT, DR. MARGARET, Women's Medical College, Philadelphia, Pa.
SWETT, DR. FRANCIS H., Duke University Medical School, Durham, N. C.
TASHIRO, DR. SHIRO, Medical College, University of Cincinnati, Cin- cinnati, O.
TAYLOR, Miss KATHERINE A., Cascade, Washington Co., Md.
TAYLOR, WILLIAM R., University of Michigan, Ann Arbor, Mich.
TENNENT, PROF. D. H., Bryn Mawr College, Bryn Mawr, Pa.
THATCHER, MR. LLOYD E., Canton, N. Y.
TINKHAM, Miss FLORENCE L., 71 Ingersoll Grove, Springfield, Mass.
TRACY, PROF. HENRY C., University of Kansas, Lawrence, Kans.
TREADWELL, PROF. A. L., Vassar College, Poughkeepsie, N. Y.
TURNER, PROF. C. L., Northwestern University, Evanstown, 111.
UHLEMEYER, Miss BERTHA, Washington University, St. Louis, Mo.
UHLENHUTH, DR. EDUARD, University of Maryland, School of Med- icine, Baltimore, Md.
UNGER, DR. W. BYERS, Dartmouth College, Hanover, N. H.
VAN DER HEYDE, DR. H. C., Galeria, Corse, France.
VISSCHER, DR. J. PAUL, Western Reserve University, Cleveland, O.
WAITE, PROF. F. C., Western Reserve University Medical School, Cleveland, O.
WALLACE, DR. LOUISE B., Spelman College, Atlanta, Ga.
WARD, PROF. HENRY B., University of Illinois, Urbana, 111.
WARREN, DR. HERBERT S., Department of Biology, Temple University, Philadelphia, Pa.
WARREN, PROF. HOWARD C., Princeton University, Princeton, N. J.
WENRICH, DR. D. H., University of Pennsylvania, Philadelphia, Pa.
WHEDON, DR. A. D., North Dakota Agricultural College, Fargo, N. D.
WHEELER, PROF. W. M., Museum of Comparative Zoology, Cambridge, Mass.
WHERRY, DR. W. B., Cincinnati Hospital, Cincinnati, O.
44 MARINE BIOLOGICAL LABORATORY
\YHITAKER, DR. DOUGLAS M., Columbia University, New York City,
N. V.
WHITE, DR. E. GRACE, Wilson College, Chambersburg, Pa. WHITING, DR. PHINEAS W., University of Pittsburgh, Pittsburgh, Pa. WHITNEY, DR. DAVID D., University of Nebraska, Lincoln, Nebr. WIEMAN, PROF. H. L., University of Cincinnati, Cincinnati, O. WILLIER, DR. B. H., University of Chicago, Chicago, 111. WILSON, PROF. H. V., University of North Carolina, Chapel Hill, N. C. WILSON, DR. J. W., Brown University, Providence, R. I. WOGLOM, PROF. WILLIAM H., Columbia University, New York City,
N. Y.
WOODRUFF, PROF. L. L., Yale University, New Haven, Conn. WOODWARD. DR. ALYALYX E.. Zoology Department, University of
Michigan, Ann Arbor, Mich.
YOUNG, DR. B. P., Cornell University, Ithaca, N. Y. YOUNG, DR. D. B., University of Maine, Orono, Me. ZELENY, DR. CHARLES, University of Illinois, Urbana, 111.
THE RELATION BETWEEN CLEAVAGE AND TOTAL
ACTIVATION IN ARTIFICIALLY ACTIVATED
EGGS OF URECIIIS
ALBERT TYLER
(From the William G. Kerckhoff Laboratories of the Biological Sciences, California
Institute of Technology, Pasadena, California and the William G. Kerckhoff
Marine Laboratory, Corona del Mar, California)
It is generally assumed in most work on artificial parthenogenesis that cleavage and development result when the initial response of the egg to the artificial agent most closely resembles its response to the sperm. The percentage of eggs that respond in this fashion varies, of course, with the length of exposure to the artificial agent, presumably reaching a maximum for the exposure producing the highest percentage of activation.1
It would follow then that the cleavage-activation relation should be such that as the percentage of activation increases, the percentage of cleavage increases; that is, that the percentage of cleavage is directly proportional to the percentage of activation. Although this relation is practically always tacitly assumed in parthenogenesis experiments, detailed data on this point are lacking. If, however, exposures giving higher percentages of activation do not produce increasing percentages of eggs whose response is most nearly like that induced by the sperm, or if such eggs were not the ones which cleave and develop, an entirely different cleavage-activation relation might be expected. The determination of this relation is important, then, in an analysis of the factors which determine whether or not an artificially activated egg will cleave.
The variation of the percentage of activation with the length of exposure to the artificial agent is in itself a highly interesting fact, since it is not manifested in insemination of a normal batch of eggs with normal sperm. This variation may be attributed to variability in the amount of treatment necessary to activate a given egg, or, less likely perhaps, to a variation in the time at which the change produced by the activating agent reaches a given egg. Whatever its source, the way in which the percentage of activation varies with the length of exposure is useful in helping to elucidate the mechanism by which the artificial agent activates the egg.
1 Any egg in which initial developmental changes have taken place will be termed ''activated" in this paper, regardless of maturation or cleavage.
45
46
ALBERT TYLER
In the parthenogenesis experiments on Urechis eggs a unique relation between cleavage and activation was found, such that as the percentage of activation increases, the percentage of cleavage decreases. The variation of percentage activation with length of exposure was found to give a particular type of distribution curve in certain of the experiments. These results together with their interpretation are presented in detail in this report.
MATERIAL AND METHOD
The eggs used in these experiments were those of the echiuroid, Urechis catipo, described by Fisher and MacGinitie (1928). The changes undergone by the egg upon normal fertilization, and upon artificial activation, and the method used in activating the eggs were described in detail in a previous publication (Tyler, 1931). Briefly, it was found that dilutions of sea water ranging from 80 per cent to distilled water were effective in activating the Urechis eggs.2 In order to treat the eggs, a batch was transferred with as little sea water as possible to a Stender dish containing a large volume of the hypotonic solution. Samples were then removed after various intervals of time to Syracuse dishes containing normal sea water. All the usual pre- cautions in regard to contamination by sperm or foreign matter, hypertonicity, etc., were taken.
TARLE I
Unfertilized Eggs Treated with Distilled Water, Temperature 21.8° C.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
|
min. |
per cent |
t>fr cent |
|
0.05 |
. 57.0 |
32.4 |
|
0.08 |
95.0 |
10.0 |
|
0.17 |
99.6 |
0.6 |
|
0.25 |
100.0 |
0.0 |
|
0.33 |
100.0 |
0.05 |
|
0.42 |
100.0 |
0.1 |
|
0.50 |
100.0 |
0.0 |
|
0.67 |
100.0 |
0.0 |
|
0.83 |
100.0 |
0.0 |
|
1.00 |
100.0 |
1.5 |
|
1.50 |
100.0 |
0.0 |
|
2.00 |
100.0 |
0.0 |
|
3.00 |
100.0 |
0.0 |
|
4.00 |
100.0 |
0.0 |
|
5.00 |
99.0 |
0.0 |
2 Eighty per cent sea water, for example, is made up of eight parts sea water and two parts distilled water. The sea water used was always taken at the same height of tide.
CLEAVAGE-ACTIVATION RELATION
47
The percentages of cleavage and of activation were based on counts of at least three hundred eggs; frequently, especially for very low or very high percentages of activation, a much larger number were counted.
It was shown that two types of activated eggs appear as a result of the treatment. One type is characterized by initial changes which are indistinguishable from those induced by the sperm. In this type the breakdown of the germinal vesicle, the rounding out of the inden- tation, the elevation of the membrane, and the extrusion of polar bodies occur in very much the same manner as when the egg is fertilized by a sperm. The time relations for these various changes, allowing for the time of exposure, compare very closely with the time schedule of the same events in the fertilized egg. However, in spite of the remarkable similarity in behaviour of this type of artificially activated egg to that of the fertilized egg, none of the eggs divide.3
The other type of artificially activated egg departs widely in its behaviour from that of the normal fertilized egg. The only visible
TABLE II
Unfertilized Eggs Treated with 20 Per Cent Sea Water, Temperature 22.0° C.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
|
min. |
per cent |
per cent |
|
0.08 |
25.0 |
69.2 |
|
0.17 |
58.1 |
58.7 |
|
0.25 |
92.0 |
31.4 |
|
0.33 |
98.8 |
17.3 |
|
0.50 |
100.0 |
3.1 |
|
0.67 |
100.0 |
1.0 |
|
0.83 |
100.0 |
4.2 |
|
1.00 |
100.0 |
0.5 |
|
1.17 |
100.0 |
0.1 |
|
1.33 |
99.5 |
2.2 |
|
1.50 |
100.0 |
0.1 |
|
1.67 |
100.0 |
0.0 |
|
1.83 |
100.0 |
0.0 |
|
2.00 |
100.0 |
0.0 |
|
2.50 |
100.0 |
0.0 |
|
3.00 |
100.0 |
0.0 |
|
3.50 |
100.0 |
0.0 |
|
4.00 |
100.0 |
0.0 |
|
5.00 |
100.0 |
0.0 |
|
7.00 |
100.0 |
0.1 |
|
10.00 |
100.0 |
0.3 |
|
15.00 |
100.0 |
0.6 |
|
20.00 |
100.0 |
1.5 |
|
40.00 |
100.0 |
0.0 |
3 In only three cases were eggs with two polar bodies seen to divide. The three eggs proceeded only as far as the two-cell stage.
48
ALBERT TYLER
change that occurs in this type of egg within the first three-quarters of an hour after treatment is the dissolution of the germinal vesicle. The egg remains indented, no membrane elevation occurs, and no polar bodies are extruded. After that time the eggs begin to round up, and lift off membranes, but no polar bodies appear. Practically all of the eggs of this type divide, the time of first division varying from one hour and twenty minutes to about three hours. The eggs which cleave and develop are thus the ones which show a poor initial response to the treatment. In what follows, then, the percentage of cleavage is practically identical with the percentage of "poorly activated eggs," and the data on percentage of cleavage and of activa- tion will also show the relation between the percentage of "imper- fectly' and of "perfectly" activated eggs for various strengths of hypotonic solutions.
THE VARIATION OF PERCENTAGE OF ACTIVATION AND OF CLEAV.M.I.
WITH LENGTH OF EXPOSURE FOR VARIOUS
DILUTIONS OF SEA WATER
Treatment icith Distilled Water
The action of distilled water is extremely rapid in causing activation of the eggs. After 3 seconds' treatment, 57 per cent of the eggs
TABLE III
Unfertilized Eggs Trailed -cilh 30 Per Cent Sea Water, Temperature 22.1° C.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
|
min. |
per rent |
per cent |
|
0.17 |
3.8 |
37.5 |
|
0.33 |
59.8 |
13.8 |
|
0.50 |
99.7 |
1.4 |
|
0.67 |
99.6 |
0.9 |
|
0.83 |
100.0 |
(i.l |
|
1.00 |
100.0 |
0.6 |
|
1.33 |
100.0 |
0.4 |
|
1.67 |
' 98.1 |
3.1 |
|
2.00 |
99.2 |
0.7 |
|
2.50 |
99.0 |
0.5 |
|
3.00 |
99.3 |
0.8 |
|
3.50 |
98.5 |
1.6 |
|
4.00 |
83.6 |
2.3 |
|
4.50 |
86.1 |
3.2 |
|
5.00 |
81.8 |
11.1 |
|
6.00 |
86.7 |
1.0 |
|
8.00 |
100.0 |
0.0 |
|
10.00 |
100.0 |
0.1 |
|
15.00 |
100.0 |
0.0 |
|
20.00 |
100.0 |
0.0 |
|
40.00 |
100.0 |
0.0 |
CLEAVAGE-ACTIVATION RELATION
49
become activated, and after 15 seconds all of the eggs are activated. The results of one series of exposures are given in Table I. Another series run in the same manner gave quite similar results. The per- centage of the activated eggs that cleave (column three in the table) is seen to drop very rapidly as the percentage of activation increases. Thus, when 100 per cent activation is obtained, there is practically no cleavage.
Cytolysis sets in after 2 minutes' exposure and reaches 90 per cent at 5 minutes' treatment. The activated eggs in that range are somewhat abnormal in appearance, having a relatively wide membrane and forming blisters over the surface so that polar bodies are often indistinguishable.
Treatment with Twenty Per Cent Sea Water
The action of 20 per cent sea water is less rapid than that of distilled water. The results of one series are given in Table II.
TABLE IV
Unfertilized Eggs Treated with 40 Per Cent Sea Water, Temperature 22.0° C.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
|
min. |
per cent |
per cent |
|
0.17 |
0.0 |
0.0 |
|
0.33 |
52.1 |
31.4 |
|
0.50 |
94.0 |
13.5 |
|
0.67 |
97.8 |
2.3 |
|
0.83 |
99.7 |
0.9 |
|
1.00 |
99.9 |
0.1 |
|
1.33 |
100.0 |
0.1 |
|
1.67 |
100.0 |
0.2 |
|
2.00 |
99.6 |
0.0 |
|
2.50 |
99.0 |
2.4 |
|
3.00 |
98.8 |
1.1 |
|
3.50 |
98.3 |
1.6 |
|
4.00 |
97.5 |
3.2 |
|
5.00 |
85.3 |
6.4 |
|
7.00 |
88.0 |
0.9 |
|
10.00 |
99.9 |
0.4 |
|
15.00 |
100.0 |
0.0 |
|
20.00 |
100.0 |
0.0 |
|
40.00 |
100.0 |
0.0 |
Fifteen seconds longer treatment is required to give 100 per cent activation than for the distilled water. The rise in percentage of activation with time of exposure is again seen to be accompanied by a drop in cleavage. No exceptions are seen in the first part of the table and the ones occurring in the latter part are of small magnitude. Cytolysis sets in after 4 minutes' exposure and reaches 70 per cent 4
50
ALBERT TYLER
after 15 minutes and 90 per cent after 20 minutes. Increasing num- bers of abnormal eggs of the type described above are found in that range. Two other series of experiments were run, and closely similar
results obtained.
TABLE Y
Unfertilized Eggs Treated with 45 Per Cent Sea Water, Temperature 21.0° C.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
Volume in M3 X 10 -» |
|
min. 0.17 |
per cent 0.2 |
Per cent 0.0 |
|
|
0.33 |
11.5 |
50.0 |
|
|
0.50 |
39.0 |
29.2 |
|
|
0.67 |
81.3 |
10.6 |
|
|
0.83 |
92.9 |
4.8 |
|
|
1.00 |
98.6 |
1.4 |
8.12 |
|
1.33 |
100.0 |
0.2 |
|
|
1.67 |
100.0 |
0.0 |
|
|
2.00 |
99.5 |
0.0 |
8.90 |
|
2.50 |
98.3 |
0.3 |
|
|
3.00 |
96.4 |
0.8 |
9.51 |
|
3.50 |
87.3 |
2.2 |
|
|
4.00 |
65.7 |
4.0 |
10.10 |
|
5.00 |
34.4 |
20.5 |
10.45 |
|
6.00 |
21.2 |
26.2 |
10.92 |
|
8.00 |
7.9 |
22.2 |
11.49 |
|
10.00 |
6.9 |
50.0 |
11.92 |
|
15.00 |
0.8 |
43.0 |
12.95 |
|
20.00 |
1.3 |
51.6 |
13.51 |
|
40.00 |
3.3 |
8.3 |
13.96 |
Treatment icith Thirty Per Cent Sea Water
With 30 per cent sea water the percentage of activation rises less rapidly than with 20 per cent. The results again show that as the percentage of activation increases, the percentage of cleavage de- creases. Table III gives the results of one series. The percentage of activation shows a slight drop after about one and one-half minutes' exposure which becomes quite marked at 4 to 6 minutes' exposure. But as the activation drops, the cleavage is seen to rise, so that at 5 minutes' exposure, where the activation has dropped to 82 per cent, the cleavage has risen to 11 per cent. The activation then rises again to 100 per cent and the cleavage drops to zero.
Cytolysis sets in after 6 minutes' treatment and reaches 50 per cent after 40 minutes. The abnormal eggs referred to above again appear in this range of exposures.
Four other series of experiments were run, at temperatures ranging from sixteen to twenty-three degrees, and very similar results obtained.
CLEAVAGE-ACTIVATION RELATK )N
51
The inverse relation between cleavage and activation was evident in each series. If every case in which an increase (or decrease) in activation accompanied by an increase (or decrease) in cleavage to the extent of at least one per cent is considered an exception, then out of a total of eighty-one dishes there are seven exceptions.
FIG. 1. Variation of percentage activation (open circles), percentage cleavage (solid circles) and mean volume of eggs (continuous curve) with length of exposure to 45 per cent sea water. Data of Table V.
Treatment with Forty Per Cent Sea Water
The percentage of activation for eggs treated with 40 per cent sea water rises less rapidly than for eggs treated with any of the preceding dilutions. Table IV gives the results of one series of exposures. The activation is seen to rise rapidly to 100 per cent, drop more slowly to 85 per cent and return again to 100 per cent. The percentage of cleavage of the activated eggs decreases as the activation increases, and increases as the activation drops. The inverse relation between cleavage and activation is thus again clearly shown, only one exception occurring in the table, namely at the three minute exposure, where a drop in activation is followed by a drop in cleavage greater than one per cent.
At the 8 minutes' exposure there is 4 per cent of cytolysis, which increases to about 30 per cent for the 40 minutes' treatment. The abnormal eggs again occur in this range.
52
ALBERT TYLER
Three other series were run with 40 per cent sea water, totaling forty-seven dishes. Out of these a total of four exceptions of magni- tude greater than one per cent were obtained.
TAHLK VI
Unfertilized Eggs Treated with 50 Per Cent Sea Water, Temperature 21.5° C.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
Volume in MJ X 10-s |
|
min. 0.17 |
per cent 0.1 |
per cent 100.0 |
|
|
0.33 |
2.0 |
80.1 |
|
|
0.50 |
43.2 |
40.6 |
|
|
0.67 |
91.0 |
21.6 |
|
|
0.83 |
96.7 |
10.6 |
|
|
1.00 |
99.2 |
5.3 |
7.83 |
|
1.33 |
99.8 |
0.7 |
|
|
1.67 |
100.0 |
0.4 |
|
|
2.00 |
100.0 |
0.4 |
8.72 |
|
2.50 |
100.0 |
1.2 |
|
|
3.00 |
90.8 |
26.9 |
9.29 |
|
3.50 |
93.4 |
21.9 |
|
|
4.00 |
86.0 |
39.2 |
9.72 |
|
4.50 |
68.3 |
63.2 |
|
|
5.00 |
33.9 |
76.2 |
10.10 |
|
6.00 |
34.9 |
80.0 |
10.46 |
|
8.00 |
18.8 |
94.7 |
11.08 |
|
10.00 |
7.1 |
83.3 |
11.42 |
|
15.00 |
0.5 |
40.0 |
12.20 |
|
20.00 |
0.4 |
79.0 |
12.72 |
|
40.00 |
1.5 |
61.3 |
12.85 |
Treatment with Forty-five Per Cent Sea Water
The results obtained with 45 per cent sea water differ in two respects from those obtained with the preceding dilutions of sea water. These are, first, that the percentage of activation returns practically to zero after its initial rise to 100 per cent, and second, that very little cytolysis sets in.
In Table V the results of one series of experiments are presented. The rate of increase in activation is slower than with the preceding dilutions. After 8 minutes' exposure a few of the eggs become cyto- lysed and the amount of cytolysis reaches 5 per cent after 40 minutes.
The inverse relation between cleavage and activation is quite evident in the table and is illustrated graphically in Fig. 1. The "exceptions" generally occur in the dishes showing low percentages of activation. Similar results were obtained in three other series of experiments run with 45 per cent sea water. Out of a total of seventy-
CLEAVAGE-ACTIVATION RELATION
53
three dishes examined, sixteen exceptions were found, all of them in dishes showing less than 8 per cent activation.
The increase in activation occurs much more rapidly than the decrease. This may readily be seen in the graph (Fig. 1), where the percentage of activation plotted against time gives a skew curve. The probable interpretation of this result will be presented later. But in connection with the activation-time curve, it is of interest to
90
BO
-tl-
II-J ~50
20
50%
FIG. 2. Variation of percentage activation (open circles), percentage cleavage (solid circles) and mean volume of eggs (continuous curve) with length of exposure to 50 per cent sea water. Data of Table VI.
present here the curve showing the increase in volume with length of exposure to the 45 per cent sea water. The data from which the curve was drawn are given in Table V. Each point represents the average of the volumes of three eggs. The measurements of the diameters were made with a Filar ocular micrometer. With this micrometer measurements accurate to 0.1 per cent may be obtained. However, the variations in volume, for the data presented here and below, ranged as high as 5 per cent. This is probably due to the rapid change in volume that is taking place as the measurements are made and to the variability of the eggs. The volume measurements are being repeated on a larger scale and by means of a cinematograph in order to obtain accurate data for an analysis of the swelling process itself. But even the relatively rough data presented here will be shown to be useful in an analysis of the activation-time curves obtained in these experiments.
54
ALBERT TYLER
The swelling curve of Fig. 1 shows that the eggs continue to increase in volume even after the percentage of activation begins to drop. The curve itself is of the exponential type, the slope continually decreasing. In other words, the increase in volume occurs less rapidly as the time of exposure increases.
TAHI.K VI I Unfertilized Eggs Treated with 55 Per Cent Sea Water, Temperature 212° C.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
Volume in M1 X 10-* |
|
min. 0.17 |
per cent 0.4 |
per rent 100.0 |
|
|
0.33 |
0.9 |
100.0 |
|
|
0.50 |
10.0 |
53.3 |
|
|
0.67 |
38.4 |
37.4 |
|
|
0.83 |
70.3 |
29.4 |
|
|
1.00 |
89.5 |
17.6 |
7.74 |
|
1.33 |
96.2 |
' 9.6 |
|
|
1.67 |
98.4 |
2.0 |
|
|
2.00 |
99.9 |
1.0 |
8.73 |
|
2.50 |
97.8 |
1.6 |
|
|
3.00 |
93.3 |
5.6 |
9.32 |
|
3.50 |
88.8 |
7.1 |
|
|
4.00 |
85.6 |
12.4 |
9.57 |
|
5.00 |
66.4 |
21.3 . |
9.82 |
|
6.00 |
64.1 |
21.0 |
10.11 |
|
8.00 |
47.3 |
16.6 |
10.49 |
|
10.00 |
13.9 |
15.4 |
10.89 |
|
12.00 |
1.7 |
44.2 |
|
|
15.00 |
1.1 |
70.6 |
11.41 |
|
20.00 |
0.0 |
0.0 |
11.63 |
|
40.00 |
2.2 |
90.9 |
11.84 |
Treatment with Fifty Per Cent .Sea Water
The results obtained with 50 per cent sea water are quite similar to those obtained with 45 per cent sea water, except that the increase in activation occurs more slowly and practically no cytolysis occurs in any of the dishes. The increase in volume of the eggs in 50 per cent sea water is also somewhat slower than for those exposed to 45 per cent sea water, and the equilibrium volume attained is, of course, smaller.
Table VI contains the results of one series of experiments and a set of volume measurements (means of three eggs) obtained at dilterent times. The data are presented graphically in Fig. 2. The results again bear out the inverse relation between cleavage and activation. It is interesting to note that the irregular rise in the activation curve
CLEAVAGE-ACTIVATION RELAT I ON
55
at the three and one-half minutes' exposure is accompanied by a drop in cleavage. The exceptions occur only in the last four points of the graph, where the percentage of activation is low. A total of eighty-two dishes in five series of experiments gave sixteen exceptions all of the same type illustrated here.
The activation shows a drop to practically zero after its initial rise
100 «
Minutes
FIG. 3. Variation of percentage activation (open circles), percentage cleavage (solid circles) and mean volume of eggs (continuous curve) with length of exposure to 55 per cent sea water. Data of Table VII.
to 100 per cent, as in the preceding case. The rise in activation is again seen to occur more rapidly than the subsequent decrease, giving the skew activation-time curve shown in Fig. 2. The volume curve for the eggs in the 50 per cent sea water is of the same type as obtained with the preceding dilution.
Treatment with Fifty-five Per Cent Sea Water
The results of one series of experiments with 55 per cent sea water and the volume data for the same dilution are presented in Table VII and Fig. 3. The data again show a decrease in cleavage as the activation increases and an increase in cleavage as the activation decreases. Four series of experiments totaling seventy-four dishes gave twelve exceptions — chiefly at low percentages of activation.
56
ALBERT TYLER
The activation-time curve is of the same shape as that obtained in the preceding case, hut it is shifted slightly to the right, so that the time required for the maximum percentage of activation and for the return to zero per cent activation is longer than with 50 per cent sea water.
The swelling curve (Fig. 3) shows that the volume continues to increase after the percentage of activation has reached a maximum. It is also of the exponential type in which the rate of increase in volume decreases with time.
Treatment with Sixty Per Cent Sea Water
The results obtained with 60 per cent sea water again differ from the preceding only in the time relations of activation and cleavage and the volume curve. The data is given in Table VIII, and graphi- cally represented in Fig. 4.
TAHLK VIII
Unfertilized Eggs Treated with 60 Per Cent Sea Water, Temperature 22.3° C.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
Volume in M3 X 10-* |
|
m i n . |
ffr i '-ill |
per cent |
|
|
0.17 |
0.1 |
0.0 |
|
|
0.33 |
1.0 |
0.0 |
|
|
0.50 |
1.0 |
50.0 |
|
|
0.67 |
13.4 |
33.3 |
|
|
0.83 |
30.3 |
25.8 |
|
|
1.00 |
61.3 |
25.9 |
7.88 |
|
1.33 |
73.1 |
13.7 |
|
|
1.67 |
88.7 |
7.7 |
|
|
2.00 |
99.4 |
0.4 |
8.68 |
|
2.50 |
100.0 |
0.1 |
|
|
3.00 |
100.0 |
0.2 |
9.16 |
|
3.50 |
97.6 |
2.9 |
. |
|
4.00 |
88.2 |
9.3 |
9.45 |
|
5.00 |
85.7 |
5.5 |
9.69 |
|
6.00 |
75.0 |
30.6 |
10.01 |
|
7.00 |
42.4 |
46.5 |
|
|
8.00 |
16.0 |
61.5 |
10.16 |
|
10.00 |
14.7 |
81.2 |
10.49 |
|
12.00 |
9.5 |
37.1 |
|
|
15.00 |
5.1 |
72.3 |
10.94 |
|
20.00 |
0.0 |
0.0 |
11.10 |
|
40.00 |
0.0 |
0.0 |
11.11 |
CLEAVAGE-ACTIVATION RELATION
57
The cleavage-activation relation shows up quite clearly. Out of eighty-nine dishes in five series of experiments, fourteen relatively unimportant exceptions were obtained.
ioor
60%
Minutes
FIG. 4. Variation of percentage activation (open circles), percentage cleavage (solid circles) and mean volume of eggs (continuous curve) with length of exposure to 60 per cent sea water. Data of Table VIII.
The activation-time curve shows a slight shift to the right when compared with the preceding ones, but its asymmetry is still quite evident.
The swelling curve is of the same type as in the preceding cases but approaches a lower equilibrium volume.
Treatment with Sixty-Jive Per Cent Sea Water
Sixty-five per cent sea water gives results which differ from the preceding in the same direction as the results obtained with the 60 per cent sea water differ from those obtained with 55 per cent. One series of experiments and a set of mean volumes are shown in Table IX and Fig. 5.
The inverse relation between cleavage and activation is evident in spite of certain relatively large irregularities. From three series of experiments seven relatively large and eight minor exceptions were obtained out of a total of fifty-nine dishes.
58
ALBERT TYLER
TABLK IX Unfertilized Eggs Treated with 65 Per Cent Sea Water, Temperature 22.0°
C.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
Volume in M3 X 10-* |
|
min. 0.17 |
per cent 0.0 |
per cent 0.0 |
|
|
0.33 |
0.2 |
0.0 |
|
|
0.50 |
0.0 |
0.0 |
|
|
0.67 |
0.7 |
40.0 |
|
|
0.83 |
6.4 |
36.3 |
|
|
1.00 |
13.6 |
11.8 |
7.61 |
|
1.33 |
77.8 |
20.8 |
|
|
1.67 |
87.6 |
13.4 |
|
|
2.00 |
92.0 |
10.2 |
8.32 |
|
2.50 |
96.5 |
6.1 |
|
|
3.00 |
98.6 |
2.1 |
8.73 |
|
3.50 |
92.5 |
6.7 |
|
|
4.00 |
71.9 |
22.5 |
9.25 |
|
5.00 |
49.0 |
56.7 |
9.56 |
|
6.00 |
21.8 |
46.9 |
9.49 |
|
7.00 |
20.3 |
51.0 |
|
|
8.00 |
13.8 |
42.4 |
9.88 |
|
10.00 |
13.7 |
34.9 |
10.15 |
|
12.00 |
6.0 |
40.9 |
|
|
15.00 |
3.9 |
15.3 |
10.50 |
|
20.00 |
0.2 |
100.0 |
10.55 |
|
40.00 |
0.0 |
0.0 |
10.55 |
The activation-time curve (Fig. 5) is asymmetrical as in the pre- ceding cases, but it shows a slight shift to the right.
•
-
'
j|
•
ii-s tso
10-
9-0- 30
65%
Minutes
I i<. 5. Variation of percentage activation (open circles), percentage cleavage -<>li<| circle^ ami mean volume of eggs (continuous curve) with length of exposure in <o |.IT cent sea water. Data of Table IX.
CLEAVAGE-ACTIVATION RELATION
59
The volume increase is slower than for the eggs in more dilute sea water, and approaches a lower asymptotic value.
Treatment with Seventy Per Cent Sea Water
The results of one series run with 70 per cent sea water are tabulated in Table X. Figure 6 shows that trend of the data graphically.
TABLE X
Unfertilized Eggs Treated with 70 Per Cent Sea Water, Temperature 212° C.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
|
min. |
per cent |
per cent |
|
0.17 |
0.4 |
57.1 |
|
0.33 |
0.8 |
50.0 |
|
0.50 |
2.5 |
55.2 |
|
0.67 |
2.4 |
29.8 |
|
0.83 |
4.1 |
30.2 |
|
1.00 |
4.6 |
29.5 |
|
1.33 |
6.4 |
20.3 |
|
1.67 |
20.4 |
19.5 |
|
2.00 |
46.1 |
10.2 |
|
2.50 |
96.2 |
9.1 |
|
3.00 |
100.0 |
2.3 |
|
3.50 |
97.5 |
10.9 |
|
4.00 |
69.2 |
19.7 |
|
4.50 |
51.6 |
35.7 |
|
5.00 |
31.6 |
55.5 |
|
6.00 |
16.7 |
65.2 |
|
8.00 |
7.4 |
82.9 |
|
10.00 |
13.3 |
76.3 |
|
15.00 |
12.7 |
85.8 |
|
20.00 |
4.6 |
83.3 |
|
40.00 |
0.3 |
66.7 |
No serious divergence from the cleavage-activation relation is evident. Two series of experiments totaling thirty-seven dishes gave seven minor variations.
The activation-time curve (Fig. 6) is again decidedly asymmetrical. It is displaced to the right, so that the return to zero per cent activation requires a longer exposure than in the preceding cases.
The volume data are not presented for this or for the succeeding dilutions of sea water. The volume increase proceeds more slowly, of course, and reaches a smaller equilibrium volume with increasing concentrations of sea water.
60
ALBERT TYLER
100
•
•
•
40
a. 30
.-
6 . , 7 8
Mi nutes
12
FIG. 6. Variation of percentage activation (open circles) and percentage cleav- age (solid circles) with time of exposure to 70 per cent sea water. Data of Table X.
Treatment icith Seventy-fire Per Cent Sea Water Table XI and Fig. 7 contain the results of one series of experiments
with 75 per cent sea water.
TABLE XI
Unfertilized Eggs Treated with 75 Per Cent Sen \\',iter, Temperature 20.8° C.
|
Length of Exposure |
Activation |
. .f Ai tiv.it'M Eggs |
|
min. |
per cent |
/vr cent |
|
0.17 |
0.0 |
0.0 |
|
0.33 |
0.0 |
0.0 |
|
0.50 |
0.0 |
0.0 |
|
0.67 |
0.0 |
0.0 |
|
0.83 |
0.0 |
0.0 |
|
1.00 |
0.0 |
0.0 |
|
1.33 |
3.3 |
71.4 |
|
1.67 |
8.2 |
50.0 |
|
2.00 |
33.9 |
45.9 |
|
2.50 |
83.2 |
I'). 5 |
|
3.00 |
86.2 |
32.3 |
|
3.50 |
90.0 |
28.4 |
|
4.00 |
79.3 |
|
|
5.00 |
47.8 |
59.2 |
|
6.00 |
30.2 |
65.0 |
|
8.00 |
7.4 |
61.3 |
|
10.00 |
1.2 |
66.7 |
|
15.00 |
0.1 |
40.0 |
|
20.00 |
0.0 |
0.0 |
CLEAVAGE-ACTIVATION RELATION
61
As before, the percentage of cleavage varies inversely with the percentage of activation, although the difference between the maximum of activation and the corresponding minimum of cleavage is not as great as in the cases listed above. Two series of experiments totaling thirty-one dishes gave four exceptions.
lOOr
Minutes
FIG. 7. Variation of percentage activation (open circles) and percentage cleav- age (solid circles) with time of exposure to 75 per cent sea water. Data of Table XI.
For the activation-time curve (Fig. 7), the time to reach a maximum is longer than in the preceding case, but the drop to zero per cent occurs sooner. However, the maximum value reached is only 90 per cent activation as compared with 100 per cent in the previous cases. The curve itself is still asymmetrical.
Treatment with Eighty Per Cent Sea Water
Eighty per cent sea water generally fails to give more than one to two per cent activation. In one series of experiments, however, an exceptionally high percentage of activation was obtained. The results are given in Table XII and Fig. 8.
It is readily seen from the data that practically every increase (or decrease) in activation is accompanied by a decrease (or increase) in cleavage, bearing out the inverse relation between cleavage and activation.
62
ALBERT TYLER
The activation-time curve reaches its maximum as quickly as for the 75 per cent sea water, but that is undoubtedly due to the higher temperature at which this series was run. The activation curve does not return to zero, but maintains a relatively high percentage of activation and a correspondingly high percentage of cleavage.
TABLE XII Unfertilized Eggs Treated with 80 Per Cent Sea Water, Temperature 22.5° F.
|
Length of Exposure |
Activation |
Cleavage of Activated Eggs |
|
min. |
per cent |
per cent |
|
0.17 |
0.0 |
0.0 |
|
0.33 |
0.0 |
0.0 |
|
0.50 |
0.0 |
0.0 |
|
0.67 |
0.0 |
0.0 |
|
0.83 |
0.2 |
100.0 |
|
1.00 |
1.4 |
42.8 |
|
1.33 |
6.6 |
31.2 |
|
1.67 |
14.5 |
30.8 |
|
2.00 |
58.2 |
26.8 |
|
2.50 |
94.1 |
21.5 |
|
3.00 |
95.7 |
17.6 |
|
3.50 |
98.8 |
3.5 |
|
4.00 |
98.7 |
8.3 |
|
5.00 |
81.6 |
57.8 |
|
6.00 |
92.5 |
37.1 |
|
8.00 |
80.7 |
48.6 |
|
10.00 |
65.5 |
59.1 |
|
15.00 |
64.1 |
38.2 |
|
20.00 |
62.5 |
48.8 |
|
40.00 |
47.6 |
64.2 |
THE VARIATION OF PERCENTAGE OF ACTIVATION WITH VOLUME FOR VARIOUS DILUTIONS OF SKA WATER
When (he percentage of activation is plotted against the mean volume attained by the eggs at different lengths of exposure, a curve is obtained which is much more symmetrical than the activation-time curve. Figure 9 shows five curves of that type, for 45, 50, 55, 60 and 65 per cent sea water. The percentages of activation were plotted in each case against the volumes attained at corresponding times of exposures, the volumes being taken from the smooth curves.
The activation-volume curves of Fig. 9 approach in shape the normal distribution curve. The individual curves have the same abscissa but the ordinates are raised successively for each dilution of
i water. It can readily be seen that even with the same coordinates
CLEAVAGE-ACTIVATION RELATION
63
the curves would not coincide; but their divergence is no greater than would be expected when one considers the statistical nature of the activation values and the errors involved in the volume measurements. Moreover, there are probably injury factors operative in the lower concentrations of sea water that are not present in the higher concen- trations, as indicated by the cytolysis obtained in 45 per cent sea water.
FERTILIZATION OF "OVER-EXPOSED" EGGS
The activation-time curves for concentrations of sea water above 40 per cent are seen to rise to a maximum of about 100 per cent activation and then drop off to zero.
100
90
80
It
60
2 &50
1 i
o -2 <t o
I
«£ 30
20
80%
QlOOCXX
Minutes
age
FIG. 8. Variation of percentage activation (open circles) and percentage cleav- (solid circles) with time of exposure to 70 per cent sea water. Data of Table XII.
The eggs which do not respond before the "optimum exposure" is reached may be termed "under-exposed" unactivated eggs, and those which do not respond upon longer exposures may be termed "over-exposed" unactivated eggs.
The failure of the "over-exposed" unactivated eggs to respond to the treatment might presumably be due to an injury effect, or other change produced in the eggs. The "over-exposed" unactivated eggs as well as the under-exposed unactivated eggs were therefore insemi- nated with fresh sperm in order to determine whether they would become fertilized and produce normal embryos. The results obtained
64
ALBERT TYLER
with 45 to 65 per cent sea water are given in Table XIII. The third column in the table gives the total percentages of activation obtained with the len.uths of exposure listed in column two. The fourth column gives the percentage of the unactivated eggs that become fertilized upon addition of sperm, and the fifth column, the percentage of the fertilized eggs that produce normal larvae.
45%
FIG. 9. Variation of percentage of activation with mean volume of eggs at- tained at corresponding times of exposure to 65, 60, 55, 50, and 45 per cent sea water. Ordinates raised successively for each dilution of sea water. Data from Tables Y to IX; volumes taken from the smooth curves of Figs. 1 to 5.
The unactivated eggs were transferred to a separate dish and inseminated at about 2 to S hours after treatment. Control eggs (listed in the table as 0.0 minutes' exposure) were inseminated at the same time.
The "under-exposed" unactivated eggs are not given for the 55 and the 60 per cent sea water. In the other three cases the "under- exposed" unactivated ei^s ^ho\v practically 100 per cent fertilization and a high percentage of normal embryos. The "over-exposed" eggs show a high percentage of fertilization in every case, comparing quite favorably with that given by the control eggs. The percentage of normal embryos obtained varies considerably, but is quite as good as that obtained from the controls, except for the 45 per cent sea water. I lo\\<-\ c] -, in the latter case a relatively large percentage of the eggs were pol\>perniic.
CLEAVAGE-ACTIVATION RELATION
65
TABLE XIII
Insemination of" Under-Exposed" and "Over-Exposed" Unactivated Eggs
|
Concentration of Sea Water |
Length of Exposure |
Ai t iv;iti<m |
Fertilization |
Normal Embryos |
|
per cent |
min. |
f>?r i'fnt |
per cent |
per cent |
|
0.0 |
• |
95 |
40 |
|
|
0.17 |
0.0 |
99 |
50 |
|
|
1.50 |
100.0 |
— • |
— |
|
|
45 |
10.00 |
5.7 |
70 |
20 |
|
15.00 |
0.0 |
60 |
2 |
|
|
20.00 |
0.3 |
60 |
5 |
|
|
40.00 |
0.5 |
60 |
5 |
|
|
0.0 |
— |
98 |
25 |
|
|
50 |
0.17 |
0.1 |
100 |
60 |
|
2.00 |
100.0 |
— |
— |
|
|
15.00 |
0.5 |
80 |
70 |
|
|
0.0 |
— |
65 |
70 |
|
|
55 |
2.00 |
100.0 |
— . |
. — . |
|
20.00 |
0.0 |
50 |
65 |
|
|
0.0 |
|
100 |
100 |
|
|
2.75 |
100.0 |
— |
— |
|
|
60 |
7.00 |
42.4 |
100 |
100 |
|
10.00 |
14.7 |
100 |
95 |
|
|
20.00 |
0.0 |
100 |
100 |
|
|
0.0 |
|
100 |
65 |
|
|
0.17 |
0.0 |
100 |
100 |
|
|
65 |
3.00 |
99.0 |
— |
— |
|
7.00 |
28.1 |
100 |
40 |
|
|
10.00 |
18.3 |
90 |
75 |
|
|
20.00 |
0.0 |
99 |
50 |
The results show that the "over-exposed" unactivated eggs are still capable of becoming fertilized, even though a shorter exposure would have resulted in every egg becoming activated upon return to
normal sea water.
DISCUSSION
1 . Variation of Rate of Increase in Activation with Dilution of Sea-Water
It is evident from the results presented above that the factors causing activation are brought into action more quickly, the lower the concentration of the sea water used for the treatment. In the dilute sea water the egg swells due to intake of wrater. The volume increase also occurs more quickly, the lower the concentration of the sea water in which the eggs are allowed to swell. This parallel be- haviour suggests that volume increase in the dilute sea water may be
5
66 ALBERT TYLER
used as a basis for an interpretation of the results presented above; but this is not meant to imply that water-intake alone is responsible for the activation of the egg.4
2. Activation-Time Curces
For concentrations of sea water ranging from 45 to 75 per cent the percentage of activation was seen to rise rapidly to a maximum and then fall off more slowly. In terms of volume change this means that when the egg is in a definite volume range it will become activated upon return to normal sea water, but before or after passing through that volume range the egg does not become activated upon return to normal sea water. This volume range is evidently well below the equilibrium volume, since the eggs continue to swell after the time of exposure giving the maximum activation. The reason why a range of volumes rather than one definite volume is specified will be indicated below. There is considerable variability in the time at which different eggs pass through the same volume range when a given batch is exposed to a given dilution of sea water. Thus some of the eggs will have reached the volume range from which return to normal sea water results in activation before the others have entered that range. Correspondingly, some of the eggs will have passed through that volume range while the others are still in it. Let us term the volume range resulting in activation the "optimum volume range." The percentage of eggs passing through a given volume range at a given time will depend on the kind of variability shown by the eggs. If the variability of this material is expressed by the normal distribution curve, then we would expect the variability in the percentage of eggs passing through the " optimum volume range " to be expressed by that type of curve only if the increase in volume were a linear function of the time of exposure. But the volume increase is a logarithmic function of time, the rate of swelling continu- ally decreasing with time of exposure. The eggs therefore enter the "optimum volume range" more rapidly than they leave it. Thus the variation in the percentage of eggs passing through the "optimum volume range" with time of exposure should be expressed by a skew distribution curve with its mode displaced to the left. In other words, the variation of percentage activation with time of exposure should give a skew curve, since the percentage of eggs passing through the "optimum volume range" is by definition identical with the percentage of activation. This is in fact the type <>t curve that is
4 The change in hydrogen ion concentration, for example, might be an important factor. It ranged from pH 8.2 for the sea water to pi I 7.1 for the distilled water used.
CLEAVAGE-ACTIVATION RELATION 67
obtained when percentage of activation is plotted against length of exposure (Figs. 1 to 8).
The reason for assuming a range of volumes rather than one definite optimum volume results from the following consideration. The maximum of the activation-time curve is at 100 per cent activation. This means that all of the eggs must be in such a condition after a certain time of exposure that removal to normal sea water at that time results in every egg becoming activated. But the volume measurements show that the eggs vary in the time of exposure at which a given volume is reached. Therefore, if we adhere to the volume interpretation we must assume that a range of volumes, at least as great as the variation in volume of the individual eggs, is effective in causing activation upon return to ordinary sea water. The time of exposure at which all of the eggs are in that "optimum volume range" then results in 100 per cent activation.
On this basis the more rapid swelling obtained with progressively lower concentrations of sea water should cause a shifting of the activation-time curve to the left proportional to the increase in rate of swelling and likewise a shortening of the time range of activation. The results presented above show that this is in general true. But with extreme dilutions of sea water (40 per cent to distilled water) the drop to zero per cent activation does not occur. This is probably due to a secondary effect as indicated by the fact that there is a tendency for the activation to drop (see Tables II, III, IV), but as cytolysis sets in a second rise in activation (of an abnormal type) takes place.
One should also expect, according to the volume interpretation, that the concentration of sea water in which the equilibrium volume of the eggs is within the "optimum volume range" should give an activation-time curve that does not drop. This is presumably approached by the 80 per cent sea water (Table XII and Fig. 8).
3. Activation- Volume Curves
If the variation of the percentage of activation with time of exposure is correlated with the variation in volume of the eggs attained at corresponding times of exposure, then the percentage of activation plotted against mean volume should give a normal distribution curve, which should be identical for the various dilutions of sea water. The results show that this is roughly true. The curves obtained with various dilutions of sea water (Fig. 9) are quite symmetrical when compared with the activation-time curves. The probable reasons for the failure of the various curves to be exactly identical have been given above.
68 ALBERT TYLER
The expectation of a normal distribution curve for percentage activation plotted against mean volume is based on the assumption that the variation in volume of the eggs, at each time of exposure considered, is expressed by the normal probability curve. This is the type of variation that is generally assumed for biological material in the absence of further information. To obtain such information in this case it would be necessary to measure the volumes of a large number of eggs at various times of exposure. This has not been done on a large enough scale and accurately enough to determine whether the chance law holds for the volumes at every exposure used, but the measurements obtained on untreated eggs indicate that their variation in volume is of that type.
4. " Over-Exposed" Unactiva ted Eggs
It has been shown that the over-exposed unactivated eggs obtained with solutions ranging from 45 per cent to 65 per cent sea water can still be fertilized and may produce normal embryos. This may be taken to mean that the eggs have not been irreversibly affected by treatment with these dilutions of sea water. Consonant with this fact is the observation previously reported, that no visible changes aside from the swelling are seen to occur in the treated eggs while in the dilute sea water. It is also in accord with the result that the time for the initial stages (e.g. polar body extrusion) of the artificially activated eggs is comparable with that of the fertilized eggs only if allowance is made for the time of treatment.
It is evident then that no developmental changes occur in the egg while in the hypotonic solution, but that activation is initiated by the return to normal sea water after a definite time of exposure (or after a certain amount of water has been taken in). The question may therefore be raised as to why a longer exposure fails to evoke a response in the egg upon return to normal sea water when a shorter one does. If the egg were found to be injured by the longer exposure this question might be more readily answered. But the data presented here show that this is not so. The question bears directly on the mechanics of activation. With the data available we can only answer by restating the result in the following terms — that a definite change (enabling the egg to become activated upon return to normal sea water) is produced in the egg by the intake of an amount of water within a certain range, but that the change is reversed when more water is taken in. In other words, by the difference in behaviour upon return to normal sea water, an egg in the optimum exposure range must be intrinsically different from an egg in the earlier or later ranges, and by the similarity
CLEAVAGE-ACTIVATION RELATION- 69
in behaviour upon return to normal sea water, an egg in the earlier range of exposures must be intrinsically the same (neglecting the manifest difference in volume) as an egg in the later range; hence the change produced must be reversed.
The return to original condition of eggs that have been allowed to swell in dilute sea water has also been noted in eggs of Nereis (Just, 1930) and eggs of Arbacia (McCutcheon and Lucke, 1926). But in neither of these cases is it stated whether activation is obtained at shorter exposures.
The ability of eggs that have been "over-exposed" to butyric acid to become fertilized has been noted by Moore (1916) for Arbacia, Just (1919) for Echinarachnius, and Lillie (1921) for Strongylocentrotus. But in these cases the cleavage and development were stated to be
abnormal.
5. Cleavage- Activation Relation
The inverse relation between the percentage of cleavage and the total percentage of activation may now be interpreted in a similar way provided we introduce a "sub-optimum volume range" on both sides of the "optimum volume range." The justification of this arises from a consideration of the results reported in a previous publication (Tyler, 1931). It was shown that the activated eggs that extrude both polar bodies practically never divide, even though the response of that type of egg to the treatment is outwardly indistinguishable from the response of the egg to the sperm. Only the eggs that produce no polar bodies were the ones to cleave, but such eggs were shown to respond in a relatively very slow and abnormal fashion to the treatment in respect to the breakdown of the germinal vesicle, rounding out of indentation, and membrane elevation. Such is the type of result one would expect from a "sub-optimum" treatment. In terms of volume change this "sub-optimum" exposure would be obtained in a "sub- optimum volume range." Practically no eggs of that type are obtained at the time of exposure giving 100 per cent activation, but they occur in increasing numbers to either side of that exposure time. Since, at the time of exposure giving 100 per cent activation, all of the eggs are assumed to be in the "optimum volume range," the "sub-optimum volume range" must occur on each side of the former.5
Thus, when a batch of eggs is treated with dilute sea water, the eggs will pass through a "sub-optimum volume range" both before and after entering the "optimum volume range." At relatively short
5 The "sub-optimum volume range" must evidently be shorter than the range of variability of the volumes of the eggs, since 100 per cent cleavage (with 100 per cent activation) is never obtained for any given exposure.
70 ALBERT TYLER
times of exposure, then, one would expect most of the activated eggs to be within the "sub-optimum volume range," and so give a high percentage of cleavage (of the activated eggs). But with longer exposures as the total activation increases one would expect more and more of the eggs to enter the "optimum volume range" and so give a low percentage of cleavage. The results would then be reversed upon passing through the second "sub-optimum volume range" with longer exposures.
This leads to a relation between percentage of cleavage and percentage total activation that is identical with that described in the text.
This interpretation can be tested in a much better fashion by following the volume changes of individual eggs in various dilutions of sea water and noting their behaviour when removed to normal sea water after having been allowed to swell to various volumes. Such experiments are now in progress.
The results reported here have an important bearing on what is generally termed the "optimum treatment" in parthenogenesis experiments. It has generally been assumed that the treatment producing the highest percentage of activation (similar to that pro- duced by the sperm), and of cleavage and development is the optimum treatment. But in Urechis it has been shown that the treatment that is optimum for activation is not so for cleavage and development. Thus, if one wishes to produce the most parthenogenetic development, the length of exposure used is different from that which would be chosen if one wished to produce the highest percentage of eggs whose initial response to the treatment was most similar to that induced by the sperm. It is preferable, I think, to term the latter the optimum treatment, for the reasons stated above. The failure of eggs receiving the optimum treatment to divide is probably connected with insuffi- cient chromatin (since all such eggs extrude two polar bodies and are left with the haploid number of chromosomes). It should be possible then to produce cleavage in such eggs by suppressing the polar divi- sions. This is somewhat difficult to accomplish without initiating other changes in the eggs, but the resu'ts obtained thus far indicate that suppression of the polar divisions of the "optimally" stimulated eggs results in cleavage.
The inverse relation between percentage of cleavage and percentage of activation appears then to depend on the fact that only the "poorly activated eggs" which extrude no polar bodies are the ones to divide. Thus the extent to which this relation is general for eggs of various forms will probably depend on whether or not the eggs that extrude
CLEAVAGE-ACTIVATION RELATION 71
both polar bodies divide. In eggs of the sea urchin type, where the polar bodies are extruded in the ovary and where cleavage is apparently possible with the haploid number of chromosomes, we might not expect this relation to hold.
In eggs of Thalassema neptuni, which, from the descriptions are very similar to the Ureclris eggs, artificial activation by means of isotonic solutions has been reported by Hobson (1928). The varia- tions of percentage of activation and of percentage of cleavage c are presented for several short series of exposures, but Hobson thinks ' that the results show an increase in cleavage with increase in activation. However, he notes (pp. 73 and 74) that the maximum of cleavage often fails to coincide with the maximum of activation, when both composition of medium and length of exposure are varied.
SUMMARY
1. The rate of increase in percentage activation of Urechis eggs with hypotonic sea water is shown to decrease as the concentration of sea water used is increased from distilled water to 80 per cent sea water.
2. The rate of increase in volume also decreases with increased concentration of sea water.
3. For dilutions of sea water ranging from 75 per cent to 45 per cent, the activation passes through a maximum (usually 100 per cent) and then returns to zero per cent with longer exposures. For lower concentrations of sea water the return to zero per cent is not obtained, but a high percentage of activation is maintained. With 80 per cent sea water the return to zero per cent activation also does not occur.
4. The activation-time curves for 75 per cent to 45 per cent sea water are of the form of skew distribution curves, rising rapidly to 100 per cent activation and falling more slowly to zero per cent.
5. The activation-volume curves are presented for 65 per cent to 45 per cent sea water and are of the form of a normal probability curve. They are roughly identical for the various dilutions of sea water.
6. Practically every series of experiments shows an inverse relation between the percentage of total activation and percentage of cleavage (of the activated eggs) ; so that as the percentage of activation increases with time of exposure, the percentage of cleavage decreases, and when
6 Hobson's total activation does not include cleavage. It is not stated in the paper whether the percentage of cleavage is that of all the eggs or of the activated eggs, though it seems to be the former. When the data of his tables is recalculated on this basis, there are thirteen cases in which an increase (or decrease) in activation is accompanied by an increase (or decrease) in cleavage and six cases in which the in- verse relation holds.
72 ALBERT TYLER
the percentage of activation decreases with exposure the percentage of cleavage increases.
7. The over-exposed unactivated eggs are still capable of fertiliza- tion and of producing normal embryos in spite of the fact that a shorter exposure would have resulted in their becoming activated upon return to normal sea water.
8. The variation in rate of activation with concentration of sea water, the type of activation-time curves, the activation-volume curves, and the fertilization of over-exposed eggs are shown to be interpretable on the basis of volume change occurring in the dilute sea water, a definite volume range being optimum for activation. The cleavage-activation relation is shown to be the outcome of the previously reported result that only the "poorly activated" eggs divide, and its interpretation, based also on the exposures producing such eggs, involves the assumption of a "sub-optimum volume range" on both sides of the- optimum.
BIBLIOGRAPHY
liMiMK, \V. K. AND (>. E. MAC i.iMMK, 1(>28. A new Echiuroid Worm. Ann. and Mag. Xat. Hist., Ser. 10, 1: 199.
FISHKU, \V. K. AND G. E. MAC I.IMIIK, 1<)2,X. Tin- Natural History of an Echiuroid Worm. Ann. anil Mag. Xul. Hist., Ser. 10, 1: 204.
HOBSOX, A. D., 1928. The Action of Isotonic Salt Solutions on the Unfertilized Eggs of Thalassema neptuni. Writ. Jour. Exper. Biol., 6: 65.
|i ST, E. I\.. \')\(). The Fertili/ation Reaction in Echinarachnius parma. III. The nature of the activation of the egg by butyric acid. Biol. Bull., 36: 39.
Ji ST, E. I-'.., 1930. ilydration and Dehydration in the Living Cell. III. The fertilization capacity of Nereis eggs after exposure to hypotonic sea water Protoplnsmu, 10: 24.
LILLIE, E. K., 1921. Studies of Eertili/at ion. IX. On the question of superposition of fertili/ation on parthenogenesis in Strongylocentrotus purpuratus. Biol. Hull., 40: 23.
McCu'K in <>N.. M., AND I.ICKK, B., 1926. The Kinetics ot ( >smoiic Swelling in Liv- ing Cells. Jour, di'ii. Physiol., 9: 0(>7.
MOORE, C. K., 1916. On the Superposition of Kertiii/ation on Parthenogenesis.
Hi«l. Hull., 31: IS 7.
, ALBKKI. I'MI. The Production of Normal Embryos by Artificial Partheno- genesis in the Hrliiumi.l. I'rechis caupo. Biol. Bull., 60: 187.
THE OCCURRENCE OF MELANOPHORES IN CERTAIN
EXPERIMENTAL WOUNDS OF THE GOLDFISH
(CARASSIUS AURATUSY
GEORGE MILTON* SMITH ANATOMICAL LABORATORY, SCHOOL OF MEDICINE, YALE UNIVERSITY
While studying in the goldfish the repair of experimental wounds, crushes, burns, and fractures, it became apparent that melanophores developed in the wounds a few days after the trauma and later de- generated and thus disappeared. Not alone did these melanophores occur directly at the site of the injury, but not infrequently in the corium of adjacent areas and even in remote cutaneous regions. In none of these places were black pigmented cells seen by a previous low- power microscopic examination of the living fishes used for the experi- ment, nor were melanophores of the corium noticeable by high magnifi- cation in sections of tissue removed from the region of the wound at the time of trauma. The appearance of pigmented cells at the very point of injury seemed to indicate a role of importance for melanophores of this fish, from the viewpoint that these cells functioned in the pro- cesses of repair and, not unlikely, in the mechanism of body defense.
As results of different experiments were found to be uniform, only a few are here reported in detail as illustrative.
Experiment 1. Goldfish, 8 cm. long from snout to base of tail, kept in still water tank, supplied by current of air. Temperature of Wc.ter 78° F.
Oct. 28, 1930. Transverse incision was made with a cataract knife through a single ray of caudal fin, near the upper edge of middle part of this fin. Incision penetrated tissues over both surfaces of fractured ray.
Oct. 30. Overlying the ray near the fracture are a few scattered melanophores with irregular processes (Fig. 1). Tissues overlying the fracture are cedematous and difficult to photograph for this reason. There are a few small points of hemorrhage near the fractured frag- ments.
Oct. 31. A large number of melanophores, in places interlacing, surround the proximal and the distal fragment of the fractured ray as if to encapsulate the fragments (Fig. 2).
1 Aided by grant from Blossom Fund.
73
74 GEORGE MILTON SMITH
Xov. 1. Active degeneration of melanophores has begun with pigment granules lying free in tissue spaces (Fig. 3).
Xov. 5. Degeneration of all melanophores in the region of the fracture, with many small pigment masses scattered throughout the field.
Xov. 8. Entire region of fracture, somewhat whitish and trans- lucent, shows no more evidence of pigment.
In the following experiment multiple injuries were produced.
Experiment 2. Two goldfishes, 7 cm. from snout to base of tail were placed in a tank of still water fed with a current of air. The temperature of the water was gradually raised from 70° F. by heating over a period of three days to 84° F.
Sept. 23, 1930. In both fishes eight different regions were clamped with an artery forceps each for 15 seconds. The points clamped were as follows: right and left opercuhun, both pectoral, both ventral, the anal and the caudal fins.
Sept. 25, 2 P.M. One fish shows early pigmentation by melano- phores in caudal fin, the second fish has melanophores in the right ven- tral fin. Pigmentation is slightly distal to crush.
Sept. 26, 3 P.M. Three days after trauma, both fishes show pig- mentation by melanophores at all eight points crushed. The pigmen- tation is a marked one due to the large number of melanophores present in the crushed zones and neighboring tissue.
EXPLANATION OF PLATE 1
FIGS. 1-4. Experimental linear fracture by incision of a ray of caudal fin of goldfish, the injury including all tissues directly overlying fracture. Letters A and B indicate site of fracture. All photomicrographs taken from the same living fish ana-sthetised with chloretone 1-201)0. Magnification X 90. Temperature of water 78°-80° F.
Kn;. 1. Two days after injury. A few melanophores have appeared in the u-dematous tissue near the fracture, A.B.
FIG. 2. Three days after injury. Numerous melanophores appearing as single cells or interlacing cells at the line of fracture, A.B.
IK.. 3. Four days after injury. Degeneration of melanophores at the site of fracture A.B. has begun. Small black pigment masses from degenerated cells lie scattered among living melanophores.
In.. 4. Five days after injury. Degeneration of melanophores at the site of fracture A.B. is complete, scattered pigment debris remains in the field. Final dis- appearance of all pigment on the eighth day after injury.
I n.. 5. Inter-radial tissue of caudal fin showing melanophores distributed near capillaries marked A, B, C, D. X 60. Fresh tissue removed from goldfish near an area crushed eight days previously. Fish outdoors exposed to sunlight,
FH,. 6. Irregular areas of pigmentation of melanophores developing on the surface of the body of a goldfish injured by removal of all body scales five days previously. Photograph made from living fish anaesthetised with chloretone. Size, two-thirds normal.
MELANOPHORES IN THE ( .< )U >KISI 1
75
I'l \TE I
2.
3.
Lr
5.
6.
76 GEORGE MILTON SMITH
Sept. 28. There is evidence of degeneration of melanophores at all crushed points. Temperature, 90° F.
Oct. 5. One fish is entirely clear of degenerated pigment granules at crushed point. The second shows a few black granules in the wound of the caudal fin.
Oct. 6. In both fishes all evidence of pigment formed of de-
ici.iU'd melanophores has disappeared at all eight points crushed. Thus these two fishes injured by crushing at eight separate points have shown, with temperature of water between 84° F. and 90° F., an intense pigmentation by melanophores at crushed points, a subsequent de- generation of melanophores, and a complete disappearance of all pig- ment detritus all in the course of 13 days.
Experiment 3. In this experiment, involving injury to the right operculum, 30 goldfishes, about seven cm. in length, were used. These were divided into three groups of ten. Each group was placed in a separate tank of running water in the laboratory. Fishes in Tank 1 were operated on by resecting one third of the right operculum by a straight vertical cut with scissors. Fishes in Tank 2 received a simple vertical crush for fifteen seconds of the middle of the right operculum. Fishes in Tank 3 were tirst crushed tor 15 seconds by a clamp placed vertically in the mid-point of the operculum and all opercular tissue distal to the clamp was resected. Tank 1 — Fishes (simple excision of one half of the right operculum) showed melanophores in the margin of the wound three days after operation. At first only a few such cells, but in the following two or three days there were many present. Evidence of degeneration of melanophores was noted in places as early as two days after their first appearance. Complete disappearance of black degenerated pigment from the wounded area varied between 3 and 9 days. Fishes in Tanks 2 and 3 with more severe injuries of the operculum showed a beginning accumulation of melanophores in the injured operculum also three days after trauma. The entire disap- pearance of pigment from the wound in fishes in Tank 2 (vertical crush of operculum) varied between (> to 15 days after appearance of melano- phores. In Tank 3 (fishes with crushed and partially resected right operculum) the eruption of melanophores at the injury occurred also three days after injury, bin the final disappearance of pigmented debris varied between 9 and 16 days. One fish in Tank 1 and four fishes in Tank 2 showed slight pigmentation by melanophores of the opposite uninjured operculum, arising when the accumulation of melanophores on the injured side was well developed. Melanophores in the area of secondary pigmentation degenerated and disappeared those in the experimentally injured right operculum.
MKLANOPHORES IN THE GOLDFISH
SUMMARY
The onset of cutaneous pigmentation by melanophores in three different types of wound of the operculum carried on simultaneously in three different tanks of running water at 76° F. was uniformly between the third and fourth day after trauma. The final disappearance of pigment of degenerated melanophores of the wound area varied between 6 and 19 days after injury. In some fishes the accumulation of melano- phores noted at the wound was relatively slight; in others the black pigmentation caused by large numbers of melanophores was intense and remained over a longer period.
Fishes operated on during the cold winter months and kept in tanks of cold running water (43° F.) did not show at wounded areas such a rapid development of melanophores as described in the preceding ex- periment. Further, pigmentation of wounds under winter temperature extended over longer periods. Thus, in nine fishes with right oper- culum crushed for 15 seconds with an artery clamp placed at the middle of the operculum, followed by excision of opercular tissue distal to the operculum, the following results were obtained: An eruption at the injured operculum in all nine fishes occurred between 13 to 16 days after injury; pigmentation had cleared up by degeneration of melano- phores in only three fishes two months after injury, with temperature of water at 53° F. It took approximately one more month (tempera- ture 53°-56° F.) for four more fishes to clear; the remaining two fishes cleared at the end of still another month or four months from the date of injury, when the temperature of the water had gradually risen to 61° F. The longest period of pigmentation in a wound of this series represented approximately 110 days from the date of the first appear- ance of melanophores.
It became of interest to learn whether or not in fishes kept in very cold water, an appearance of melanophores after trauma could be temporarily inhibited, to appear for the first time when such fishes were changed back slowly to more favorable warmer temperatures. A number of experiments were done along these lines.
Experiment 4. A goldfish, seven cm. in length, was placed in a tank of still cold water supplied by a current of air, the water varying in temperature between 42° F. and 45° F. The tank was set up in a refrigerator arranged with a double window, admitting ample daylight. It was found advisable to accustom the experimental fishes gradually to cold. By using several submerged electric lights at the beginning of the experiment and turning these off as desired, the temperature of the water could be lowered slowly without endangering the life of the fish.
78 GEORGE MILTON SMITH
Oct. 14, 1930. A small incision was made with a cataract knife in the caudal tin of this goldfish dividing transversely a single ray near the upper margin of the tin. Examination of melanophores at four day intervals negative for an entire month. Temperature 42° to 46° F.
Nov. 14. Temperature in tank raised slowly so as to reach 66° F. on Nov. 16th.
Nov. H). Numerous melanophores appeared for the first time at fracture and along injured ray distal to this. No other black pigmenta- tion noted.
Nov. 25. Slight pigmentation by melanophores of tip of tail and also along the margin of dorsal fin. Large accumulation of melano- phores at fracture.
Nov. 27. Active degeneration of melanophores at fracture and other pigmented regions.
Nov. 29. Fish under dissecting microscope shows no pigment masses either at site of experimentally fractured ray or at the second- ary points of black pigmentation of tail or dorsal tin. All melano- phores have disappeared by a process of degeneration.
Fishes kept in a dark chamber, excluding all light, developed mel- anophores in wounds as promptly as did controls kept in daylight.
Experiment 5. Two goldfishes, seven cm. in length, with crushed right operculum and caudal fin, kept in a dark chamber in a tank of still water at 64° F., supplied by air current, were taken out of this chamber to be examined for the first time after injury on the fifth day. Many melanophores were present in crushed regions. At the same time, two control fishes, injured on the same day in a similar way, kept in a tank of equal size at the same temperature but exposed to labora- tory daylight, exhibited, also for the first time, a large number of melan- ophores at the two crushed points. Twenty-three days after injury, one fish contained in the dark chamber and both controls were clear of pigment; the second fish in the dark chamber showed no melanophores in the injured operculum, although a few small masses of degenerated pigment masses still remained in the caudal fin.
The production of a second injury in a healed wound frequently, but not always, caused another eruption of melanophores. Refractur- ing a single ray at the same point, especially where the previous healing had left a wide whitish translucent area, did not produce a second crop of melanophores. The very simple injury of making a longitudinal slit in the caudal fin did not call forth melanophores either at the time of the first injury or with repeated incisions at the same point.
The irregular topographic distribution of melanophores following
MELANOPHORES OK THE GOLDFISH 79
trauma was seen particularly well in experiments where the scales on both sides of the body were totally removed.
Experiment 6. Nov. 1930. Three goldfishes, A, B, C, measuring 8, 7, 5 cm. in length respectively, kept in a heated tank of still water 76° F., supplied by air current, were operated on under chloretone anaesthesia (1-2000). All scales of the body were removed with forceps in all three fishes.
Dec. Four days after operation, melanophores appeared in ir- regular groups at various points on both sides of the body. The two larger fishes, A and B, showed in the course of the next few days a large number of melanophores in irregular scattered patches. The patches of pigmentation by melanophores in fish B are shown in Fig. 6. The smallest fish, C, showed only a few melanophores in small, widely- scattered areas. By the end of the twelfth day degeneration of melan- ophores evoked by removal of scales had occurred in all three fishes with a disappearance of broken-down pigment material. At this time (12 days after removal of scales) each fish showed definitely a set of new young scales. Fish B successfully withstood a second complete re- moval of scales, under chloretone anaesthesia, but this time only a very few rapidly degenerating melanophores developed on the denuded surface of the body, as if the supply of pigment-forming cells for these particular surface areas were partially exhausted. When, however, on the fourth day after the second operation for removal of scales the caudal fin of this fish was crushed by clamp for 15 seconds, numerous melanophores developed three days later in the crushed tail but in no other place.
DISCUSSION AND SUMMARY
Various important problems relating to melanophores and melano- genesis appear in connection with the works of Van Rynberk (1906), von Frisch (1911), Weidenreich (1912), Asvadourova (1913), Spaeth (1913), R. Fuchs (1914), Wyman (1924), Wells (1925), Abolin (1925), Ewing (1926), Jost (1926), Bloch (1927), Cordier (1928), Becker (1930).
For the present purpose it may be of interest to recall that a number of years ago Weidenreich (1912) showed that in vertebrates the distri- bution of black pigment cells could be regarded as forming four distinct envelopes for the body. These envelopes he designated as "cutaneous, perineural, pericoelomatic and perivascular" respectively. He pointed out that whereas in some vertebrates several or all of these pigmentary envelopes were well developed, in other vertebrates one or more of these pigmentary envelopes might be found poorly developed, showing only a trace or rudiment of pigmented tissue. For example,
80 GEORGE MILTON" SMITH
in man, where there exists a. well developed cutaneous envelope of pig- mented tissue, the perineural pigmented tissue is poorly developed, presenting itself as scattered black pigment cells of the piamater and elsewhere in the brain. In fishes all pigmentary envelopes are regarded as fairly well developed.
In interpreting the meaning of melanophores following injury as seen in the above experiments on goldfish, it should be kept in mind that such melanophores may represent a perivascular or perineural type of cell developing the properties of forming pigment, rather than cells belonging strictly to a system of cutaneous melanophores. It is particularly the cutaneous or corial melanophores of fishes which have received the most study to date.
Melanophores, according to Bloch (1927) show a number of mor- phologic peculiarities in that they form processes or dendrites and have a tendency to arrange themselves in an interlacing net\vork. They exhibit in cold-blooded animals certain functional reactions which are shown by the spreading or the contraction of the intracellular masses of pigment granules. These reactions are changes which have their origin in nervous, actinic or hormonal stimuli; and they may also be produced by mechanical, chemical and electric means.
Ever since the description of melanophores in fishes by Siebold (1861), many investigators have contributed to the morphology of this subject. The works of Ballowitz (1912-16) on the different types of chromatophores (i.e., the melanophores, xantho or erythrophores, ^iianophores and their various combinations forming what he desig- nated as chromatic organs) have largely laid the basis for our present knowledge of pigment cells in fishes. This author also demonstrated histologically the innervation of melanophores in fishes.
The experimental observations of Pouchet (1876) showed a rela- tionship between cutaneous melanophores in fishes and the sympathetic nervous system. It remained, however, for von Frisch (1911, 1912), in a series of important experiments, to demonstrate in fishes a con- traction center for cutaneous melanophores in the front part of the medulla, and a secondary center in the spinal cord. Further, he ex- plained the pathways by which impulses pass from brain through pig- ment motor nerve fibers to the sympathetic system and from here by means of the peripheral nerves not only to the melanophores but also to other chromatophores of the skin.
In general, the function of melanophores has been variously in- terpreted. In addition to the view that cutaneous pigmentation and pigment changes represent color adaptation to environment, the pur- pose of cutaneous pigment has been thought to lie in its protection ot
MELANOPHORES IX THE GOLDFISH 81
deeper tissues against injurious solar rays. The migration of retinal pigment granules as it applies to vertebrates and arthropods is thought by Parker (1906) to be a mechanism calculated to protect the receptive organs of the retina from o~\ e -stimulation by light and to improve the ' retinal images. Cutaneous pigment cells have been regarded as trans- forming light into heat energy. According to this view, as Weidenreich (1912) explains, the minute individual intracellular pigment granules of melanophores become heat bodies or Heizkorper, which distribute heat to neighboring protoplasm. Weidenreich (1912) has further suggested, because melanophores are innervated and react to optic, thermic and chromatic stimuli, that they may be regarded perhaps as sensory cells for color and warmth perception.
Cordier (1^28) believes that the formation in cells of melanin is a process of excretion as yet not well understood. The theory implies that certain toxic waste products of metabolism gain access to special cells and there become insoluble and pigmented, their toxic products being neutralized. Elimination of pigment follows slowly as if it were a process of retarded excretion. Certain clinical cases of Addison's disease and melanosarcoma have shown melanin greatly increased in cutaneous areas and present in the blood and in the urine. This has been taken to mean a profound chemical disturbance of the body as a whole and gives support to the view that a general metabolic process may ordinarily affect the production of melanin in various regions of the body.
Whatever may be the relationship to the nervous system of melano- phores resulting from trauma as seen in the present experiments on goldfish, it seems plausible from their structural arrangement in healing wounds, that such melanophores are pigmented cells which function in repair of damaged tissue. Melanophores of this kind appeared rela- tively early in the course of wound-healing when favorable warm tem- peratures were employed. They disappeared by a process of degenera- tion at the site of the wound when healing proceeded and usually when the covering of the wounded surface was nearing its completion. Whereas melanophores showed in wrounds of goldfishes within 3 or 4 days after injury when fishes were kept in water of relatively warm temperature (70°-90° F.), with fishes kept in cold water (40°-42° F.) the appearance of melanophores in wounds was retarded or even in- hibited, to appear for the first time when these fishes were returned to a warm environment. A temperature of 40° F. was found sufficient to inhibit the appearance of melanophores for a month.
Fishes kept in a dark chamber completely excluding light showed melanophores in various experimental wounds as early as did controls
6
82 GEORGE MILTON SMITH
kept under usual laboratory conditions exposed to light. Fishes kept in tanks out-of-doors and in this way exposed directly to the sunlight developed melanophores in wounds a few days later (Fig. 5). The reaction here seemed intense. In some of these fishes melanophores developed not alone at the crushed points, but also in areas adjacent to the wound and in all other fins.
When studied in a simple form of injury such as dividing trans- versely a single ray of the caudal fin, melanophores appeared first as periadventitial cells in close relation to the outer \valls of the small capillary blood vessels which covered the surface of the ray near the fracture. With an increase in numbers, the melanophores spread toward the region of the fracture and formed a network (Fig. 2) in the corium by the interlacing of the numerous irregular processes. De- generation in individual melanophores was observed as early as 24 hours after their first appearance near a fractured ray. Fixed paraffin sections of tissue with degenerating melanophores showed a moderate number of phagocytic cells containing pigment. For the most part, however, the impression was gained that the pigment detritus rested free in the tissue spaces preparatory to removal by lymphatics, or became dissolved in situ.
The actual production of melanin in cells is now generally regarded as the result of enzyme action. The important studies of Bloch (1927), advancing the views on the intracellular production of melanin by enzyme, are too well known to need repetition here. It is conceivable that in the experimental wounds of goldfish chemical changes occur locally permitting melanin to be formed in periadventitial cells ir- regularly distributed in the corium of the injured area.
Experimental wounds of goldfishes quite naturally are constantly open to infection by bacteria or parasites. Numerous bacteria and especially cocci were seen in paraffin sections of tissue from crushed operculum at various stages after injury before complete healing had occurred. When, as occasionally noted, a growth of fungus appeared in connection with experimental wounds, pigmentation by melano- phores appeared particularly intense, affecting not alone the wound but also adjacent areas. There was at times pigmentation of the fins other than the ones experimentally injured and, in rare instances, a patchy pigmentation of body scales under these circumstances. Treating such wounds for several days in succession with two per cent mercurochrome destroyed the parasites, and pigmentation of the wound with secondary pigmented areas then disappeared. The pres- ence of bacteria in wounds and the large number of melanophores present in injured areas affected with parasites, suggest a possible role l"r melanophores in the mechanism ot body defense.
MELANOPHORES IX THE GOLDFISH
Goldfishes subjected to a total removal of scales showed in the course of several days a distribution of melanophores varying in extent and intensity in different fishes. This eruption was asymmetrical, ir- regular and patchy, as if periadventitial cells capable of forming black pigment as a result of trauma or during subsequent wound regeneration, actually occupied a very irregular distribution on both sides of the body. As new scales formed in these experimentally produced scale- less fishes, melanophores disappeared by degeneration. A second total removal of scales in one of the fishes was followed by a very scanty eruption of melanophores, as if the possibility of local melanophore production in this instance were, temporarily, at least, exhausted.
Usually, but not always, a re-injury at the same point brought out a second eruption of melanophores differing but little from that which followed the primary injury.
The eruption of melanophores in experimental wounds of the gold- fishes, varying in intensity in different fishes, appears to indicate that such melanophores, probably periadventitial in origin, form in response to injury and function in the repair of injured tissues.
LITERATURE CITED
ABOLIN, L., 1925. Beeinflussung des Fischfarbenwechsels durch Chemikalien.
Arch. f. mikr. Anat. und Entwick., 104: 667. ASVADOUROVA, N., 1913. Recherches sur la formation de quelques cellules pigmen-
taires et des pigments. Arch, d'anat. micros., 15: 153. BALLOWITZ, E., 1893. Die Nervenendigungen der Pigmentzellen, ein Beitrag zur
Kenntnis des Zusammenhanges der Endverzweigungen der Nerven mit dem
Protoplasma der Zellen. Zeitschr.f. wissenschaft. Zool., 56: 673. BECKER, S. W., 1930. Cutaneous Melanoma: a Histologic Study especially directed
toward the Study of Melanoblasts. Arch. Dermal, and Syph., 21: 818. BLOCH, B., 1927. Das Pigment. Handbuch d. Haut. u. Geschlechtskrankheiten
Berlin, Vol. 1, Part 1, pp, 434-541. CORDIER, R., 1928. Les pigments melaniques et la melanogenese. Bull. Soc. Roy.
d. Sc. med. e nat. de Bruxelles, Nos. 2-7, pp. 43-57. EWING, J., 1922. Neoplastic Diseases. Philadelphia, pp. 871-890. VON FRISCH, K., 1911. Beitrage zur Physiologic der Pigmentzellen in der Fischhaut.
Arch.f. ges. Physiol., 138: 319. VON FRISCH, K., 1912. Uber farbige Anpassung bei Fischen. Zool. Jahrbuch, 32:
171. FUCHS, R. F., 1914. Der Farbenvvechsel und die chromatische Hautfunktion der
Tiere. Handbuch d. vergleich. Phys., 3: 1189. Josx, F., 1926. Die Farbzellen und Farbzellvereinigungen in der Haut des Nordsee-
fisches Callionymus lyra L. Zeitschr.f. mikr. anat. Forschung, 7: 461. PARKER, G. H., 1906. The Influence of Light and Heat on the Movement of the
Melanophore Pigment, especially in Lizards. Jour. Exper. Zool., 3: 401. POUCHET, G., 1876. Des Changements de coloration sous 1'influence des nerfs.
Jour, de I' Anat. et de Physiol., 12: 1-90, continued 113-165.
VON SIEBOLD, C., 1863. Die Siisswasserfische von Mitteleuropa. Leipzig, p. 14. SPAETH, R. A., 1913. The Physiology of the Chromatophores of Fishes. Jour.
Ex per. Zool., 15: 527.
84 GEORGE MILTON SMITH
VAN RYXBERK, G., 1906. t'ber den durch Chromatophoren bedingten Farbenwechsel
der Tiere (sog. chromatsche Hautfunktion). Ergebn. der Physiol., 5: 347. WEIDENREICH, F., 1912. Die Lokalisation des Pigmentes und ihre Bedeutung in
Ontogenie und Phylogenie der \Yirbeltiere. Zeitschr. f. Morph. u. Anthrop.,
Sonderheft 2, pp. 59-140.
WEILS, H. G., 1925. Chemical Pathology. Philadelphia, pp. 526-532. WY.MAX, L. C., 1924. Blood and Nerve as controlling Agents in the Movements of
Melanophores. Jour. Exper. Zool., 39: 73.
STUDIES ONr THE INTESTINAL FLORA OF TERMITES
WITH REFERENCE TO THEIR ABILITY TO
DIGEST CELLULOSE
ALBERT DICKMAX (From the Department of Bacteriology, University of Pennsylvania)
INTRODUCTION
Interest for a considerable time has been centered on the ability of certain organisms to derive nourishment from a wood diet, the principal constituents of which are cellulose and lignin, both resistant to the digestive action of enzymes normally present in the digestive tract of most animals. Animals such as termites, larvae of wood-boring beetles, and a bivalve, the shipworm Teredo navalis, so injurious to ships and piles, have been conspicuous for their ability to digest cellulose. In the case of Teredo navalis (Dore and Miller, 1923), digestion has been explained by the production of cellulose-digesting enzymes. The explanation in the case of termites is a more interesting one.
Microscopic examination of the intestinal content of most species of termites discloses countless numbers of Protozoa and bacteria. Careful experiments have been carried out to explore the relationships between the host and their intestinal organisms, and it has been shown conclusively that the termites are absolutely dependent upon the Protozoa present for the digestion of the cellulose in their food.
L. R. Cleveland, one of the foremost workers in this field, has shown that termites containing an intestinal fauna of Protozoa were able, under favorable conditions, to exist on a diet entirely made up of Whatman filter paper of the purest grade, and he successfully reared them upon this diet for over two years. If, however, he defaunated the termites (by incubating them at 36° C., the Protozoa were killed, but the termites were unharmed), they were unable to live upon the pure cellulose and soon died. If he re-inoculated the termites with Protozoa, however, after incubation, they were able to live indefinitely upon the filter paper (Cleveland, 1924).
BIOLOGICAL CONSIDERATIONS
Comparatively little work has been done with respect to the biological relationship between termites and the other intestinal organ- isms usually present. Hollande (1922) discusses the morphology and
85
86 ALBERT DICKMAX
reproduction in a considerable number of spirocructes which he found very abundant in the intestinal contents. Hoelling (1910), in his paper on "The Xuclear Conditions of Fusiformis termitidis," describes the morphology of fusiform bacilli studied from a number of smears of tlu' intestinal content of termites. Imms (1924) states that " Portier in\ c-n-, ited an apparent symbiosis in the case of the larva of Nonagria which lives within the stems of Typha devouring the pith. ... In the digestive tube of this larva are found great numbers of motile conidia of a fungus (Isaria), which exist among the devoured vegetable frag- ments. The conidia are always accompanied by a micrococcus which secretes an enzyme capable of dissolving cellulose. Portier states that the conidia develop at the expense of the dissolved cellulose and eventually penetrate the walls of the gut, escaping into the blood. Most of them are there attacked by phagocytes and transformed into products which serve to nourish the tissue of the host."
In an attempt to determine whether the Protozoa were entirely responsible for the digestion of cellulose in the digestive tract of termites, Cleveland (1924) studied the bacterial flora of Reticulitermes flavipes. He states that bacteria were sometimes numerous, and he attempted all known methods, aerobic and anaerobic, for isolating cellulose-digesting bacteria. One hundred attempts were made, but all results were negative, even after the cultures were more than two months old. Ten attempts to isolate cellulose-decomposing molds and actinomyretes were made and were unsuccessful. In an attempt to isolate the organisms, an inorganic, medium was made containing:
KoHPO, 1.00 gram
MgSO, 0.50 gram
KC1 0.50 gram
leSO, .01 gram
Na.\<>. 2.00 grams
H<) lOOOcc.
To this medium cellulose was added in two forms: a small piece of Whatman's filter paper and 0.5 per cent cellulose suspension. To the inorganic medium containing cellulose suspension sufficient agar was added to make a solid medium. Incubation was apparently 36° C.
Cleveland (1(^2S) observed that all families of termites harbored many spirocruetes which he thought might play a role in the digestion o| cellulose and hemicellulose. He observed millions of these, often attached to a single Proto/ofm, and easily mistaken for flagella. At tempi- to -row the spirocruetes failed and animal inoculations I n-ved negative. By feeding the termites cellulose thoroughly moist-
INTESTINAL FLORA OF TERMITES 87
ened with a 5 per cent aqueous solution of acid fuchsin, he found it possible to remove in this manner all spirocha?tes without doing any damage whatever to the Protozoa or to the termites. He concluded that the spiroclurtes play little if any role in the digestion of wood and cellulose.
In the wood-ingesting larvae of certain insects, characteristic blind sacs and diverticula of the digestive tract have been demonstrated in which, aided by the action of myriads of bacteria, food particles are held and digested. From the larvae of rose-chafers (Potosia cuprea) cellulose-digesting bacteria have been isolated in pure culture. These slender peritrichiate, anaerobic rods (Bacillus cellulosae fermentans \Yerner) are found free also in the ant hills inhabited by Potosia cuprea. The optimum temperature for fermentation was found to be 33-37° C., the minimum 21° C. The larvae are so dependent upon these intestinal organisms that the increase in weight of the larvae is determined by the temperature. If the temperature of the ant hill at the end of October goes below 21° C., the now useless taking-in of food material is sus- pended (Buchner, 1928).
EXPERIMENTAL
a. The Food of Termites
It was the purpose of the present experiment to determine whether cellulose-digesting organisms occurred in woody material upon which termites feed. The woody material from termite colonies was first examined for cellulose-digesting organisms. Some material obtained from a termite colony in March, 1929, which had been kept in a dry condition since that time (almost one year), was used to inoculate nitrate-cellulose tubes. The material contained wood particles, termite excreta and a small quantity of fine reddish clay.
The nitrate-cellulose medium was made according to the formula of Bradley and Rettger (1927). It contained:
Di Potassium phosphate 1 gram
Magnesium sulfate 1 gram
Sodium chloride 1 gram
Calcium carbonate 2 grams
Potassium nitrate 2 grams
Distilled water 1000 cc.
The cellulose was provided in the form of strips of filter paper. Tubes were kept at room temperature and incubated at 34.5° C. and 60.5° C. under aerobic and anaerobic conditions.
Ten tubes were incubated at room temperature under aerobic
88 ALBERT DICKMAX
conditions. In every one of these there was abundant growth and discoloration of paper above the liquid level. Marked discoloration appeared in all tubes in four days. Seven tubes labeled G-l to G-7 inclusive were inoculated from two of the above tubes on March 1 , 1930. When examined on March 3, 1930 discolored areas were present on the paper of all tubes. On the original tubes the discolorations were yellow-green, yellow and light brown. On the "G" tubes the predominant color was dark brown.
Three tubes inoculated and placed under anaerobic conditions on February 19, when examined on March 8, showed no marked dis- coloration and no cutting of paper at this time. (Anaerobic conditions were produced in a Mason jar, using pyrogallic acid and sodium hy- droxide.) On this date the three tubes were placed under aerobic conditions. When examined April 7, tube no. 16 showed cutting of paper at liquid level and maceration of paper below liquid level. Mold growth apparently was inhibited.
A tube no. 12 incubated at 34.5° C. under aerobic conditions on February 19, when examined on February 25 showed marked discolora- tion similar in variety and extent to that produced at room tempera- ture. The paper, however, was not cut at liquid level.
Three tubes 17, 18, and 19, inoculated and incubated at 34.5° C. under anaerobic conditions, showed no noticeable discoloration or cutting of paper when examined 18 days later. The tubes were re- moved from the incubator and placed under aerobic conditions, at room temperature. When examined 16 days later, the paper in tubes 18 and 19 was entirely cut at liquid level with no characteristic discoloration. The paper below the liquid level was entirely macerated. (The paper was probably cut before the day of examination.)
A tube incubated at 60.5° C". under aerobic conditions showed, when examined about 10 days later, a few small, isolated areas of growth on paper.
Three tubes were inoculated and incubated at 60.5° C. under anaerobic conditions on February 20. When examined on March 7, one of the tubes showed paper cut at liquid level and macerated at lower portion below liquid level. In this tube the paper was so mace- rated that upon slight shaking it fell apart into loose fibers below liquid level.
From this last tube three tubes were inoculated and incubated at 60.5° C. under aerobic conditions on March 8, three silica-gel plates inoculated from the above tube also, were incubated anaerobically at 60.5° C. On April 7, neither plates nor tubes showed discoloration or growth. Re-inoculated nitrate-cellulose tubes at 60.5° C. under anaerobic conditions showed no growth after one month.
INTESTINAL FLORA OF TERMITES 89
Woody material obtained with termites from Dr. Cory was used to inoculate a tube at room temperature under aerobic conditions. Th material was composed of wood and digested material, but no loos soil. When examined 9 days later the paper at the liquid level showe^ a discoloration which was marked above liquid level. Green discolora-\ tion was prominent. One colony 0.5 mm. in diameter produced a wine- colored discoloration. Transplants on silica-gel plates produced abundant growth.
The silica-gel was made fundamentally according to the general formula of Winogradsky (1929). Seventy-five grams of potassium silicate were dissolved in 1000 cc. of distilled water. To this was added an equal quantity of HC1 of a specific gravity of 1.10. Thirty cc. of mixture were placed in the petri dishes and set aside under cover for 24 hours. They were then immersed in running water for 24 hours. They were next washed four successive times for 24 hours each in large covered dishes of sterile distilled water. From the following inorganic salt solution (made up for 100 plates) 2 cc. were added to each silica-gel dish.
KH2PO4 1.0 gram
MgSO4 0.5 gram
NaCl 0.5 gram
FeSO4 0.01 gram
MnSO4 0.01 gram
KNO;! 3.6 grams
CaCO- 2.0 grams
Distilled water 200 cc.
The pH was adjusted to 7.2. The petri dishes were then placed in an incubator at 60.5° C. until excess moisture evaporated. Sterilized pieces of Whatman's filter paper were placed aseptically on silica-gel plates. Sterile covers were then placed over the plates. Stroke in- oculations were made.
Termites received from Dr. Cory were transferred to clean petri dishes in which were placed filter paper and the tissue paper sent with the original shipment. This was being eaten by the termites. On March 8, pellets of termite excreta which had been dropped on the tissue paper were used to inoculate two nitrate-cellulose tubes and one silica-gel plate, and kept at room temperature. (The pellets were clean and the color of tissue paper.) On March 10 no visible growth was evident in the tubes. Yellow discoloration was noticeable on the paper around the pellets in the silica-gel plates. This growth later covered the entire paper and the paper showed almost entire digestion by May 10. Nitrate-cellulose tubes inoculated from this plate showed growth
90 ALBERT DICKMAX
and digestion of paper in 5 days. A gelatinous milky-white growth with translucent areas developed on paper. Microscopic examination showed maceration of fibers with attached bacteria. The growth was characterized by the presence of mold growth which formed the . i i.itinous milky mass.
b. The Intestinal Contents of Termites
Most of the experiments with the intestinal contents were carried out with termites of the genus Reticulitermes collected at Mullica Hill, Xew Jersey, and with Tennopsis, received from Dr. Kirby, at the University of California.
Reticulitermes- Microscopic examination of intestinal contents showed besides the myriads of Protozoa, spirocrurtes ranging in length from 5 to 15 n; the smaller ones were in great abundance, and appar- ently more numerous than the Protozoa. Motile rods and filamentous rods were also present. Examinations were made with hanging drops in physiological salt solution and from smears stained with alcoholic fuchsin.
On March 31, seven inoculations were made with intestinal con- tents of seven termites, washed 4 minutes in 1- 1000 HgClo, then rinsed with sterile distilled water. The intestinal contents were squeezed out with >tcrilc forceps onto silica-gel plates. These were kept anaerobi- cally at room temperature. On April 1, sixteen termites were used as above to inoculate silica-gel plates. Eight of these were kept under anaerobic conditions and eight were kept at room temperature under aerobic conditions. On April 2, twenty termites were used as above to inoculate silica-gel plates kept at room temperature under aerobic conditions. ( )n April 1 , six termites were used to inoculate six nitrate- cellulose tubes.
When the above cultures were examined on May 12, none of the anaerobic pl.iic- -Imwed growth. Of the aerobic silica-gel plates eight inoculations showed .1 Alight mold growth, with no cutting ot paper and no distinct discoloration. Tin- remaining inoculations on plates produced no growth. One of the six nitrate-cellulose tubes showed clearly digestion of paper, with translucent areas, and microscopic examination of paper from this tube showed numerous bacteria, 1 X 0.75 /j. in si/e on the libers.
Four tubes inoculated on February 28 with Reticulitermes flavipes received from Dr. L. K. Cleveland showed no growth on March 10, and no growth was visible on April M).
1 i-nini/tsis. — The specimens were large enough to enable one to dis- sect out tin- digestive tract with sterile forceps. All termites were
INTESTINAL FLORA OF TERMITES <H
first washed for 5 minutes in 1 1000 HgCl2 and then rinsed in sterile distilled water. All inoculations were made on silica-gel plates, which were kept under aerobic conditions at room temperature.
On April 21, eight termites were used for inoculations. On April 23, two winged termites were used to inoculate silica-gel plates. On April 24, ten winged termites were used for inoculations. On April 24, eight worker termites were used as above. On May 2, six workers were used as above. On May 2, eight winged forms were used as above. On April 23, ten termites were used to inoculate silica-gel plates which were kept under anaerobic conditions. When examined on May 17, none of the plates showed cellulose-digestion or indication of growth of cellulose-digesting organisms. There was mold growth on intestinal contents of eighteen termites. The growth of molds was evidently due to the fact that treatment for 5 minutes with 1-1000 HgCl2 did not kill them.
Beckwith and Rose (1929) obtained cellulose digestion in a number of cases wThen working with intestinal contents of termites, but their results cannot be taken as conclusive, since they attempted to sterilize the termites externally by merely washing them in tincture of iodine (U.S. P.) for 45 seconds. This short exposure and the fact that small air bubbles captured between the hairs of the insect would prevent contact with the germicide in this time, would indicate that the organ- isms on the surface were not destroyed.
DISCUSSION OF RESULTS
Cleveland and others have shown that digestion of cellulose in termites is entirely dependent upon intestinal organisms. Cleveland has shown that termites are not dependent upon intestinal spirochaetes, although he has not shown that the spirochartes do not play an im- portant part in cellulose digestion.
In every experiment carried out in the present study with woody material from termite nests, abundant growths of cellulose-digesting organisms were obtained. The numerous cellulose-digesting organ- isms, wrhich were undoubtedly taken into the digestive tract with food, could not again be isolated from the intestine on the cellulose media used.
The most conspicuous organisms, with reference to numbers and bulk next to the Protozoa, as seen by microscopical examination of the intestinal contents of termites, are the spirochaetes. Since these do not grow on the usual laboratory media their true significance has not been explained.
92 ALBERT DICKMAN
SUMMARY AND CONCLUSIONS
Cellulose-digesting organisms, both bacteria and molds, are very abundant in termite nests. These organisms have been obtained in abundance from material dried for a year.
Cellulose-digesting bacteria were not isolated from the intestines of termites on the nitrate-cellulose medium of Bradley and Rettger nor on the silica-gel medium of \Yinogradsky.
True bacteria are probably of little importance in cellulose digestion
in termites.
BIBLIOGRAPHY
BECKWITH, T. D., ROSE, EDYTHE J., 1929. Cellulose Digestion by Organisms from
the Termite Gut. Proc. Soc. Exper. Biol. and Med., 27: 4. BRADLEY, L. A., RETTGER, L. F., 1927. Studies on Aerobic Bacteria Commonly
Concerned in the Decomposition of Cellulose. Jour. Bacterial., 13: 321. BUCHNER, PAUL, 1928. Holznahrung und Symbiose. Berlin. CLEVELAND, L. R., 1924. The Physiological and Symbiotic Relationships between
the Intestinal Protozoa of Termites and their Host, with Special Reference
to Reticulitermes llavipes Kollar. Biol. Bull., 46: 178. Ci EVELAND, L. R., 1925. The Method by which Trichonympha campanula, a
Protozoon in the Intestine of Termites, Ingests Solid Particles of Wood for
Food. Biol. Bull., 48: 282. CLEVELAND, L. R., 1925. The Ability of Termites to Live Perhaps Indefinitely on a
Diet of Pure Cellulose. Biol. Bull., 48: 289. CLEVELAND, L. R., 1925. The Feeding Habit of Termite Castes and its Relation to
their Intestinal Flagellates. Biol. Bull., 48: 295.
CLEVELAND, L. R., 1925. The Effects of Oxygenation and Starvation on the Sym- biosis between the Termite, Termopsis, and its Intestinal Flagellates.
Biol. Bull., 48: 309. CLEVELAND, L. R., 1928. Further Observations and Experiments on the Symbiosis
between Termites and their Intestinal Protozoa. Biol. Bull., 54: 231. DORS , \V. H., MILLER, R. C., 1923. The Digestion of Wood by Teredo navalis.
I'niv. Calif. Publ. in Zoo/., 22: 383. HOELLING, B. A., 1910. Die Kernverhaltnisse von Fusiformis termitidis. Arch.
Protistenk., 19: 239. HOLLANDE, A. C., 1922. Les Spirocht-tes des Termites; processus de division;
formation du Schizoplaste. Arch, de Zoo/. Exper. et Gen., 61: 23. IMMS, A. D., 1924. A General Textbook of Entomology. London. McBiiTH, I. G., 1916. Studies on the Decomposition of Cellulose in Soils. Soil
Science, 1: 437. WAKSMAN, S. A., CAREY, C., 1926. The Use of the Silica Gel Plate for Demonstrating
the Occurrence and Abundance of Cellulose- Decomposing Bacteria. Jour.
Bacterial., 12: 87. WINOGRADSKY, S., 1929. Etudes sur la microbiologie du sol. Ann. de I'Institut
Pasteur, 43 : 549.
THE INNERVATION OF THE STOMACH AND RECTUM
AND THE ACTION OF ADRENALINE IN
ELASMOBRANCH FISHES
BRENTON R. LUTZ
(From the Mount Desert Island Biological Laboratory, Maine and the Physiological Laboratory of Boston University, School of Medicine)