nity aia x Ferri pierseageh et uarpe* oh A Ae ratio eat fh ITP Pe at tea lene il,piesttal ie in i 7 Lgetn A jake Brot te as Trias Hes * apr anh A - 4 . ti ia try ? ata tee i iat He Hs * a} rete ; re} Nv ry ‘3 ae pals TRANSACTIONS maith of the Kentucky Academy of Science Affiliated with the A. A. A. S. VOLUME EIGHT Twenty-fifth and Twenty-sixth Meetings fe War AD By of Pohea a ai a Lat marae * a SES FB 3% -- Pole fait eee al Lexington, Kentucky 1940 TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE AFFILIATED WITH THE A.A. A.S. VOLUME EIGHT TWENTY-FIFTH AND TWENTY-SIXTH MEETINGS 1938-1939 This Volume was Edited by A. M. PETER and THE PUBLICATIONS COMMITTEE, LEXINGTON, KENTUCKY 1940 CIOEN AE ESN eS Ti! IM EMTLOTIA MD, pc. YIN aka ale A Ree can rei ete loon Se eee 3 Offices; ©1937-1939"... itch or: | tae. ok ag ee t/a alt ca Ae 4 Muntiressot the 25 thy Annuals Mieetime nm lO3 Sires... sot. ieee 5 Miemibenssrel ected ia 93 Sima cet sitet e ee ere “ind three ELA eae 6 Divistons Of eBiolosical: SGlEN GES sneer eee alte ia ete eee ee 7,34 Division), “OPiChemistry) 208... a. eke tee cee Sere oo aie 12,39 Division: of GC eology sandy Geognap lnyiie staserern selene ee eee 14,41 Mathematical Association of America, Kentucky Chapter ............. 17,44 Divaston wots Philosophy sands PSsycholooss sae eraeti en etal nen 17,44 Kentucky Psychological Association ............:.........+.+++--4--- 17 American Association of Physics Teachers, Kentucky Chapter ........ 19,42 JUDO Academy sof iS Glencen LOS Sr erriese tree) icke tri eee eae te eee 19 Minutes of sther 26 thasanmual ey Mie cri tion 5 0 eeu tee ee eee 21 Papers presented at the 26th Meeting ................ Sans Atte Sa oe 23 Louise AsiromonmiGall SOC oodesosansscesenqccueboogsecoccceoue: 42 Juimor Avexclenmmy Or Seiemce, IO) 2. .cceesscou essences cocnscesessccs 45 NUK G Keb CREne ania IRR Nene dee a NAMES aah A MONE ARE, Cua pina ee eROR ET CINE SA'S 8 ne oo 46 ae $n Memoriam : 4 S Lewis Cass Robinson, University of Kentucky Wilfred A. Welter, Morehead State Teachers College Dn e_ ; Kentucky Academy of Science OFFICERS 1937-1938 President, L. Y. Lancaster, Western State ‘Teachers College. Vice-president, Theodore Beust, University of Louisville, Deceased, October, 1937. A. C. McFarlan, University of Kentucky Secretary, Alfred Brauer, University of Kentucky. Treasurer, Julian Capps, Berea College. Representative on the Council of the A.A.A.S., A. R. Middleton, University of Louisville President Emeritus and Editor of the Transactions, A. M. Peter. Councilor for the Junior Academy, Anna A. Schnieb. 1938-1939 President, W. R. Allen, University of Kentucky. Vice-president, James L. Leggett, Transylvania College. Secretary, Alfred Brauer, University of Kentucky Treasurer, William J. Moore, Eastern State Teachers College. Representative on the Council of the A.A.A.S., A. R. Middleton, University of Louisville. President Emeritus and Editor of the Transactions A. M. Peter. Councilor for the Junior Academy, Anna A. Schnieb. AFFILIATED ORGANIZATIONS Kentucky Junior Academy of Science Kentucky Ornithological Society Kentucky Psychological Association Kentucky Section, American Association of Physics ‘Veachers Kentucky Chapter, Mathematical Association of America Kentucky Section, American Chemical Society Louisville Astronomical Society Kentucky Association of Science- Teachers MINUTES OF 25th ANNUAL MEETING; MOREHEAD STATE TEACHERS COLLEGE, May 13 and 14, 1938 The meeting was called to order and presided over by the President, Dr. L. Y. Lancaster. President H. A. Babb, of the Morehead State ‘Teachers College eave the address of welcome. Dr. A. R. Middleton gave the response. The Secretary, Alfred Brauer, gave his report which included the following: Active membership, 230. Honorary members 13. Life members, 2. An important event in the history of the Academy was the receipt in De- cember, 1937, of a check for $281.30, the amount of the L. Otley Pindar legacy. This was deposited with the Lexington Federal Savings and Loan Association, at interest. This brought the total invested endowment of the Academy to $356.30. The interest from this investment is used in the publication of The Transactions. Award of the A.A.A.S. research grants was made as follows: W. A. Welter, Morehead, Ky., fifty dollars for travel expenses in connec- tion with his Ecological and Taxonomic problem of the Licking drainage system. Gordon Wilson, Bowling Green, Ky., thirty-five dollars for preparation of a publication on “Bird Life on a Temporary Lake Produced by Overflowing Sink Holes in Western Kentucky.” John B. Loefer, Berea, Kentucky; sixty-five dollars for apparatus to be used in his study of the chemistry of excretion in Paramecium. The committees appointed by President Lancaster are as follows: MEmpersHIP: J. IT. Skinner, Charles Hire, W. A. Welter, Wm. Clay, E. N. Fergus, V. F. Payne. Pusiications: A. M. Peter, L. Y. Lancaster, Alfred Brauer, W. R. Allen, Julian Capps. Aupitinc: A. D. Hummel, Waldemar Noll, L. G. Kennamer. Procram: Alfred Brauer, L. W. Lancaster, Anna Schnieb, A. R. Middleton, Robert Hinton, Julian Capps, A. C. McFarlan. RELATIONS wiTH A.A.A.S.: A. R. Middleton, V. F. Payne, Alfred Brauer. Nominatinc: Anna Schnieb, J. S. McHargue, J. G. Black. REsoLuTions: W. R. Allen, John S. Bangson, P. A. Davies. The Resolutions Committee and members from the floor proposed the fol- lowing resolutions, which were adopted: 1. That public libraries be urged to so organize their books that a patron can be given Primary, and then Secondary books on a scientific subject. 2. That the recent establishment by the W.P.A. of a state-wide museum project merits our approval as well as the cooperation of individuals and all local agencies in the effort to build museums in our schools and towns. 3. That it be suggested to the proper authorities of the W.P.A. that they undertake a scientific collection of facts pertaining to automobile traffic ac- cidents thruout the State, to the end that future legislative and administrative efforts to reduce such accidents may rest upon factual bases. 4. That the Academy instruct the Executive Committee to study the feasibility of creating a standing State Committee whose function it shall be to promote the interests of Science in Kentucky and the more efficient function- 5 6 Kentucky Academy of Science ing of the Academy as an advisory body. It is the thought of the present com- mittee that younger scientists of the state should be worked into active partner- ship in the affairs of the community, thru this committee. 5. That the Secretary of the Academy be instructed to write the California Society for the Promotion of Medical Research, conveying to them the endorse- ment of this Academy of their program of resistance to the proposed California so-called “Humane Pound Law.” We reiterate resolutions of former years con- demning all types of anti-vivisection laws designed to destroy modern physiolog- ical and medical research. 6. That the Secretary of the Academy be instructed to write The Hon. Harold L. Ickes, Secretary of the Interior, as well as other members of the Senate Committee, that the Kentucky Academy of Science stands with other good and patriotic citizens in opposition to the enactment of S.B.3925, a bill to au- thorize the construction of a dam at Yellowstone Lake, and all similar acts proposing the conversion of natural areas in national parks. 7. ‘That the Academy express its condemnation of the general practice of school officials in assigning the teaching of Science to athletic coaches not well prepared for that specific work. 8. That the Academy seriously consider inaugurating the custom of having the president act as the annual speaker, giving an address which will adequately summarize some aspect of his particular field of interest, wide or narrow. 9. That the transactions of the Academy are worthy of much improved publicity in the newspapers of the state. 10. ‘That the Executive Committee prepare separate resolutions on the death of Theodore Beust, Vice-President 1937-1938. The following new members were added to the roll, upon recommendation of the Membership Committee and by unanimous vote of the Academy: Adams, Wm. Jr., Estill Bailey. John H., Ashland Bondurant, John H., Experiment Station, Lexington Braun, Katherine, Morehead State Teachers College Catlett, Lucille, Morehead State Teachers College Caudill, Fred, Morehead State Teachers College Chamberlain, Dr. Leo M., University of Kentucky, Lexington *Cochran, Wellington, Experiment Station, Lexington Crouch, Dr. H. B., K. S. I. C., Frankfort Davenport, F. G., Transylvania College, Lexington Diachun, Stephen, Experiment Station, Lexington Fair, L. A., Morehead State Teachers College Falls, J. D., Morehead State Teachers College Gray, Mary Campbell, Barbourville Groves, H. H., Transylvania College, Lexington Haggan, H. C., Morehead State Teachers College Hoke, R. L., Morehead State Teachers College Holtzclaw, J. D., Morehead State Teachers College Hood, M. Noka, Williamsburg Horlacher, L. J., University of Kentucky, Lexington Judd, R. D., Morehead State Teachers College Kern, Kenneth, Mayslick Kuiper, John, University of Kentucky, Lexington Lowry, S. J., Princeton Maxwell, Dr. E. S., Lexington Meece, L. E., University of Kentucky, Lexington Twenty-Fifth Annual Meeting 7 Mitchell, J. S., University of Kentucky, Lexington Moore, Amy Irene, Morehead State Teachers College Nickell, Clarence, Morehead State Teachers College Payne, Patrick M., Hazard Penix, Doris, Sharpsburg Plummer, Niel, University of Kentucky, Lexington Roofe, Dr. Paul G., University of Louisville _ Ryland, Dr. Hobart, University of Kentucky, Lexington “Saunders, J. M., Transylvania College, Lexington Seay, Maurice F., University of Kentucky, Lexington Senff, E. K., Morehead State Teachers College Sullivan, John L., Morehead State Teachers College Thompson, J. Walter, Webbville Vernon, Dr. Clarence, University of Louisville Waters, Elton, Transylvania College, Lexington Williams, Ella, Lexington Wimmer, C. R., Barbourville Wright, H. A., Transylvania College, Lexington The slate of officers selected by the Nominating Committee was elected. They are, President, W. R. Allen, University of Kentucky, Lexington Vice-President, James L. Leggett, Transylvania College, Lexington Secretary, Alfred Brauer, University of Kentucky, Lexington ‘Treasurer, Wm. J. Moore, Eastern State Teachers College, Richmond Representative on A.A.A.S. Council, A. R. Middleton, University of Louisville Instead of the usual presidential address a roundtable discussion was held on “The improvement of the Academy.” This was led by President Lancaster and resulted in the resolutions adopted at the meeting Alfred Brauer, Secretary DIVISION OF BIOLOGICAL SCIENCES ‘H. P. SturpIvANT, Chairman Jay B. Kenyon, Secretary Osmotic TOLERANCE OF CERTAIN FRESH-WATER Protozoa. John B. Loefer, Berea College. An experimental attempt was made to increase the osmotic toler- ance of Colpidium campylum, Glaucoma piriformis, Chlorogonium euchlorum, Euglena gracilis and Khawkinea halli. The organisms were grown under bacteria-free conditions in Van’t Hoff solution (Hall’s 1924 formula) to which 0.2% Bacto-tryptone and 0.02% yeast extract had been added for nutrient and to maintain the pH at 6.7. The organisms from regular stock media were transferred to a mixture of one part Van’t Hoff solution and 99 parts distilled water containing peptone. After incubating several days, a few drops of the cultures were transferred aseptically to a similar peptone medium having, however, a higher salt concentration. The procedure was repeated with each species thru a number of transfers so that each species was gradually transferred to successively higher and higher salt concentrations ranging from 1, 5, 10, 15— 100% Van’t Hoff mixture. In this way it was possible to acclimatize several 8 Kentucky Academy of Science species to osmotic pressures greater than they withstood on sudden transfer. In the course of ten to fifteen subcultures Colpidium developed an osmotic tolerance to 3.8% saline, which is greater than that of sea water (3.2-3.3%), yet on sudden transfer it had withstood only a salinity of 1.5%. Glaucoma from a regular stock medium could be changed directly to a medium with a total salinity of 1.5%; after gradual acclimatization it withstood 2.7%. The tolerance of Chlorogonium increased from 1.5 to 2.5%; Euglena developed no tolerance above 1.5%, which concentration it withstood on sudden transfer. Khawkinea was peculiar in that it was viable even after 200 hours exposure to Van’t Hoff solution having a salinity as high as 3.97%, but was never motile in concentrations above 1.5%. In the other four species motility and viability ap- peared to be directly correlated. The most pronounced morphological varia- tions observed in organisms from the higher salinities were decreased size and tendency of the contractile vacuole to disappear. LESPEDEZA PRODUCTS IN POULTRY FEEDING. J. Holmes Martin and W. M. Insko, Jr., Kentucky Agricultural Experiment Station. Ground lespedeza seed or screenings were used to replace as much as three-fourths of the meat scrap in the growing mash. Substitution of seed for half the meat scrap or of screen- ings for one-fourth of the meat scrap gave satisfactory results. Germination of the lepedeza and weed seed was reduced by passing thru the digestive tract of the growing chick. The results indicate a possibility of bringing viable weed seed to land on which poultry droppings are scattered. Trials with laying pullets indicated that lespedeza seed or screenings did not produce results quite equal to meat scrap altho egg production, egg size and hatchability were suf- ficiently high for practical results. SOME SPRING FLOWERS OF EASTERN KENTUCKY. Wilfred A. Welter, Morehead State Teachers College. About 400 feet of 16 mm film of flowers in natural color was shown to demenstrate the use of color film of flowers as an aid in teaching. THE FisHEs OF THE LickING RIVER DRAINAGE IN EASTERN KENTUCKY. Wilfred A. Welter, Morehead. This demonstration consisted of a named collection of the fishes taken in the drainage area. Most of the fishes known to occur were on exhibition; 69 species have been recorded. THE INFLUENCE OF CERTAIN ACCEssoRY GROWTH FACTORS ON NITROGEN FIx- ATION BY AZOTOBACTER. John L. Sullivan, Morehead State Teachers College. The influence of various so called accessory growth factors on nitrogen fixation by azotobacter was determined by adding different quantities of these materials to the culture media and measuring the amount of atmospheric nitrogen assimi- lated. Cysteine hydrochloride, methyl phenyl acetate, beta indole, 3-n pro- propion acid, thioglycollic acid and many other materials were used. NOTEs ON THE LIFE HIsToRY AND BREEDING HABITs OF THE EASTERN FLYING SQUIRREL. Dwight E. Soelberger. A cursory survey of the literature in 1934 re- vealed that little had been recorded concerning the habits of this unique Amer- ican mammal. Since flying squirrels were abundant in western Pennsylvania where the writer was located, work on this form appeared justified. During the Twenty-Fifth Annual Meeting 9 course of the investigation over 200 flying squirrels were observed under as natural conditions as possible. Captive animals were kept in outdoor cages for determining the period of gestation and the growth and development of the young. These observations were supplemented by observations of litters raised under natural conditions. Observations on the gliding ability, food habits, par- ental instincts, number of breeding periods per year were also recorded. EVIDENCES OF CELL MIGRATION IN THE SUPRARENTAL CorTEx. Mary Campbell. Union College. In a comparative study of fluids suitable for fixation of the en- docrine glands certain histological and cytological observations indicated a mi- gration of cells from the outer region of the zone fasciculata toward the zone reticularis. The significance of this migration to the mode of secreation of the cortex is also considered. THE EFFECT OF REMOVAL OF THE UROPYGIAL GLAND OF PIGEONS WHEN ON A. Low ViTAMIN D Ration. G. B. Pennebaker, Murray Teachers College. The gland was removed from one squab of each of the eleven pairs of nest mates, the other being kept as a control. The birds were kept on a low vitamin D ration and without access to direct sunlight. No significant difference was found in the rate of growth or general health of the two groups. But at seventy-eight days of age the operated birds had fifty-four percent heavier thymus glands, and a slightly, tho significantly, greater percentage of ash in the tibiae than the controls. LEAF ARRANGEMENT IN AILANTHUus. P. A. Davies, University of Louisville. No abstract received. . STAPHYLOCOCCI FROM CANNED OYSTERS AS A CAUSE OF Foop POISONING. Paul Majors, M. Scherago and R. H. Weaver. Univ. of Kentucky. A pure culture of Staphylococcus aureus was isolated from the liquor from a can of oysters, the contents of which was thought to have caused food poisoning in two persons. By intraperitoneal injections into kittens and by feeding human volunteers it was determined that a sterile filtrate from a 40-hour culture of this organism contains enterotoxin. This is apparently the first instance of Staphylococcus poisoning to be traced to canned food or to oysters. THE EFFECT OF SULPHANILAMIDE ON BLOOD PLATELETS AND A COMPARISON OF THE METHOD or CouNTING THEM. Lenore Fonville, M. Sherago and R. H. Weaver, Univ. of Kentucky. To carry out this study it was necessary to com- pare various methods of counting blood platelets in order to find one that was satisfactory. A direct method employing Reese and Ecker’s diluting fluid proved to be more satisfactory than any other tried. Under the conditions of this ex- periment, sulphanilamide was found to have no effect on blood platelets when administered subcutaneously and intravenously te mice and rabbits. THE MIICROFLORORA OF GLASSES FROM BEER DISPENSARIES. Marjorie Reeves Bates, R. H. Weaver and M, Scherago, Univ. of Kentucky. A survey was made of the microflora of glasses from 20 beer dispensaries in Lexington. Standard agar plates made to determine general contamination, yielded counts ranging from 200 to 52,000 colonies. The legal limit is 100. Potassium tellurite cystine blood agar plates for the isolation of diphtheroid organisms yielded typical 10 Kentucky Academy of Science colonies from the glasses from 3 dispensaries. Blood agar plates failed to indicate the presence of hemolytic streptococci on any glasses. Smears prepared by Mall- man’s method showed spirochaetes to be present on the glasses from 10 dis- pensaries. THE FAUNA OF FERMENTATION RESIDUES FROM DISTILLERIES. H. B. Crouch, Kentucky State College, Frankfort. Decomposing distillery refuse materials were collected from vats and surface pits owned by five distillery companies in Central Kentucky. All contained a variety of animal life. Among the animals found liy- ing in these materials, the following were indentified: 26 genera of Protozoa, 5 yenera of Hemiptera, 6 genera of Coleoptera, and 2 genera of Diptera. The pits were also frequented by the sandpiper bird, and at least one species of water snake. The animal life in these materials manifests several significant biotic re- lationships. Since distillery refuse is highly homogeneous and disposed of in large quantities, it provides excellent natural growth conditions for great numbers of animals, especialy saprozoites and water scavenger insects. Aside from the in- terrelationships of theses organisms within the refuse pits, they also have highly improtant significances to the transient animal life and to the living conditions in near-by streams. The drainage of the pits into the creeks and rivers provides abundance of microfauna for predacious animals. Many of the latter are like- wise consumed for food by larger and economically important aquatic animals. MORPHOGENESIS OF THE PELVIC GIRDLE OF AMBLYSTOMA PUNCTATUM. Harvey B. Lovell, Univ. of Louisville. The first visible chondrification of the pelvic girdle appeared in a larva 18 mm. long (Fig. A, D). No digits were visible on the limb bud. The rudiments of the right and left girdles were widely separated. In a 19.5 mm. larva, the pelvic cartilages had increased in size, but the two halves of the plate were still separated by a gap .17 mm. across (Fig. B, E). The femur was well chondrified, with its head in contact with the side of the puboischiac cartilage. In a 21 mm. larva, which had three digits, the pelvic girdle had undergone considerable differentiation (Figs. C, F). The right and left girdles had grown medially until they were only .045 mm. apart. The ilium had elon- gated dorsally, altho it was still not fully chondrified. The two girdles were found to be completely formed in a 24 mm. larva, altho they had not yet fused in the midline (Fig. G). The sacral rib could be indentified as a procartilage aggregation of mesenchymal cells; only a small area near the middle showed distinct matrix. The ilium was fully chondrified. In a 28 mm. larva (Fig. 1), the two girdles had fused to form a single puboischiae plate. The sacral rib was cartilaginous except at the extreme tip. The pelvic caruilages were relatively thin and weak, the puboischium being only .14 mm. thick. : Toward the close of larval development, the pelvic girdle thickens rapidly and undergoes considerable ossification of the ilium and ischium. In a 47 mm. larva beginning to metamorphose, the girdle had become 0.4 mm. thick, nearly four times the diameter of the previous case (Fig. J). This appears to be cor- related with the increased importance of the limbs in locomotion in the adult stage. Twenty-Fifth Annual Meeting 1] FIGURE 1. Pelvic girdle of Amblystoma punctatum. A, B, C, D, H, hind- limb buds of 18, 19.5, 21, and 25 mm. larvae, camera lucida x 14. D, E, F, G, I, J. sections thru pelvic girdles of 18, 19.5, 21, 24, 28, and 47 mm. larvae, camera lucida, D and E, x 26; F, G, I, and J, x 19. f, femur; 7, ilium; p, puboischium: s, sacral rib. 12 Kentucky Academy of Science A New NortH AMERICAN WATER-SNAKE OF THE GENUS NATRIX. William M. Clay, Univ. of Louisville. THE BEHAVIOR OF SOME FISHES IN A SMALL AQARIUM. Samuel L. Allen and L. Y. Lancaster, Western Kentucky State Teachers College. VARIABILITY IN THE DEVELOPMENT OF EGGS. FROM DIFFERENT HENS AND THE RELATIONSHIP OF RATE OF DEVELOPMENT TO HATCHABILITY. Joe K. Neel, Univ. of Kentucky. Eggs laid from 9:30 to 10:30 A. M. were gathered from trap nests, chilled to physiological zero, and put into modern air conditioned, forced draft incubators. The time required for the eggs to warm up to the incubation temperature was deducted from the total incubation period. Eggs of thirty-six hours incubation ranged from three to ten somites; eggs of forty-eight hours, from thirteen to seventeen somites. Eggs from hens of high hatchability per- centage showed higher rates of development than those from hens of lower hatchability percentage. Eggs from hens two years of age tended to develop more rapidly than those from hens one year of age, of the same hatchability level. ‘THE PRODUCTION OF SPINY SPORANGIA IN THE PHYCOMYCETE, ARAIOSPORA STREP: TANDRA. Harlow Bishop, Univ. of Louisville (Demonstration). In the life cycle of Araiospora streptandra, two types of sporangia appear, sporangia *without spines and sporangia with spines, in addition to the antheridia and oogonia. The significance of the spiny sporangia in the life history has long been puzzling. Both in nature and in culture they have been of rare occurrence. On a special culture medium, recently devised, they were grown in great abundance and, of special interest, to the exclusion of sporangia of the smooth type. This experi- ment offers the means for the eventual revelation of the function of these spiny sporangia. DIVISION OF CHEMISTRY A. W. Hompmercer, Chmn., J. T. Skinner, Sec. THE TOTAL ORGANIC ACIDS IN SOME PATHOLOGICAL URINES, H. E. Carswell, Univ. of Louisville. A Stupy oF FEEDS REQUIRED By GEESE. G. Davis Buckner, W. M. Insko, J. Holmes Martin, and Amanda Harms, Ky. Agricultural Experiment Station. A PLAN FOR THE CORRELATION OF HIGH SCHOOL AND COLLEGE CHEMISTRY. V. F. Payne, Transylvania College. Various altempts to give some recognition of the study of high-school chemistry to the student continuing the subject in college are mentioned. The separate course has sometimes failed because of the attitude of the university teachers in minimizing the value of the prerequisite Twenty-Fifth Annual Meeting 13 high-school chemistry. The practice of Ohio State University of examining the student who offers high-school chemistry for admission and permitting the student to enter a separate course if his score is satisfactory and requiring him to take the regular course for reduced credit otherwise is mentioned. The practice of Transylvania College of permitting students who make satisfactory examination scores on the high-school chemistry at the beginning ot the college course, to enter second-year chemistry or the second half of first- year chemistry is defended on the basis of: 1. the outcomes in Transylvania, 2. the better preparation of present-day high-school teachers, 3. the better equip- ment now available for high-school teaching, 4. the success of a similar practice in the foreign languages and 5, the stimulating effects of such practice on high- school teaching. EFFECT OF HUMIDITY ON THE GAIN-IN-WEIGHT METHOD FOR FOLLOWING THE CouRsE OF OXIDATION IN DRyING Ors. C. C. Vernon and W. W. Rinne, Univ. of Louisville. The gain-in-weight method has been found best adapted when data obtained under uniform conditions were compared and conclusions drawn from such comparison. Variations of humidity from approximately zero to 55-60 percent caused considerable variation in the weight of films of linseed oil containing some synthetic resin. When the variations were computed on the basis of the gain in weight attributed to oxidation, they were quite significant and indicated the necessity for control of humidity when using the gain-in- weight method. Only observations at the same humidity are comparable. The data presented indicated the existance of an equilibrium between the moisture in the air and the moisture in the film of oil. A STUDY OF TYPICAL HiGH SCHOOL CHEMIsTRY TExTs. P. N. Payne, Hazard. AN AUTOMATIC COULOMETER G. L. Gorley and B. T. Collins, Univ. of Louis- ville. A glass cell for the electrolysis of potassium mercuric iodide solution was suspended in such a way as to tip and break the circuit as soon as a certain weight of mercury had been deposited on the cathode. Results checked closely with a silver coulometer. On A Funcus Found GROWING IN DISTILLED WATER. Carl Johnson. Univ. of Louisville. A MEASURE OF PROGRESS IN QUANTITATIVE TECHIQUE. Ruth N. Fonaroff, Univ. of Louisville. A class in quantitative analysis determined the percentage of chlorine in a soluble chloride soon after the beginning of the term and again about 4 months later. The average time was cut from 13.4 hours. to 6.5 hours. The breakage decreased. The average class error changed from 12.0 percent to 0.76 percent. The median error was reduced from 3.5 percent to 0.70 percent. A comparison was made between the individual results and the psychological en- trance scores. THE EFFICACY OF VITAMIN D DERIVED ROM VARIOUS SOURCES, A. W. Homber- ger, Univ. of Louisville, 14 Kentucky Academy of Science DIVISION OF GEOLOGY AND GEOGRAPHY LuciEN BECKNER, Chairman R. E. SrouprEr, Secretary FossiL TRAcks IN EASTERN KENTUCKY. Wilbur Greeley Burroughs, Berea Col- lege. Human-like footprints* occur sunken into the surface of a nearly horizontal bed of Pottsville, Pennsylvanian, sandstone about eleven miles southeast of Berea. Each track has five toes spread somewhat apart, a distinct arch, and tapers back like a human foot to the heel. The tracks are about 9.5 inches from heel to end of longest toe. The width is 4.1 inches across the ball of the foot. Three pairs and four single tracks are entirely exposed, and two are partly cover- ed with solid sandstone. The tracks of one pair are nearly parallel and are 3.22 inches distant at their forward parts. Two pairs have one foot advanced, the distance from left to right heel being 18 inces. Parts of other footprints are dimly visible. The tracks extend in various directions. No imprint of front feet, tail or body markings was found. Proofs that the tracks were made in Pottsville sand are: the sand grains within the tracks are closer together than those outside the tracks, due to pressure of the creature’s feet, not to foreign matter washed in. The sandstone uprolls adjacent to each track, about two-tenths to four-tenths inch above the general surface of the stratum, where the sand was pushed outward and upward from beneath the foot. Iwo tracks are partially covered with Pottsville sandstone in situ. Photomicrographs and infra-red photographs reveal further proofs that footprints are real tracks. Irregularities in the sandstone were caused by weather- ing, features of the original sand surface, or both. Indians carved imitations of animal and human tracks in rock, but the tracks here described were made by creatures as yet unknown. LocaL Dips AND FAULTING IN KENTUCKY. William L. Russell, Owensboro. In Kentucky local dips are produced by structural deformation, stratigraphic causes, and superficial causes such as slumping and settling produced by solution of limestone. The faults of Kentucky, reverse and normal, have an east-west trend, swing- ing towards the southwest in extreme western Kentucky. The reverse faults form a system, the Rough Creek Faults system. These faults are associated with sharp folds and other evidences of compression, but the amount of thrusting de- creases to the east, and east of Grayson County no reverse faults are known. Certain features suggest that this system is the eastern extension of the Oua- chita belt of deformed rocks, which extends from Mexico to the Mississippi Em- bayment in Arkansas. There is a slight tendency for the Ouachita folds to bend to the north, in the areas close to the Embayment. The Rough Creek faults are north of a projection of the Ouachita folds, but they may be explained by * A photograph of one pair of these tracks appears in Science News Letter, October 29, 1938 (Vol.34, No.18, p.279.). The tracks appear dark in the photograph because the photographer dampened them to make them more distinct. Twenty-Fifth Annual Meeting 15 assuming that the belt of deformation bends around the Ozark uplift, as it bends around Llano uplift. Additional evidence is furnished by wells in the Missis- sippi Embayment in northwestern Tennessee, between the Rcugh Creek and the Ouachita structures, which penetrated deformed and altered Paleozoic rocks beneath the Cretaceous. THE KENTUCKY RIVER FAULT. A. C. McFarlan, Univ. of Ky. This is a zone of en echelon faulting extending from Mt. Sterling across the central and southern Bluegrass region. The structure is typically a “graben” but in places a single fault. A striking feature is the pronounced dip off the arch along the outermost fault. The topographic expression is marked and includes (a) an abrupt ter- mination of the Inner Bluegrass region and (b) the forming of Muldraughs Hi! along the southern border as a fault line scarp. Burdett Knob in Garrard County is a remnant of the Ohio and Waverly shales. The Knob constitutes a monadnock rising above the Lexington pene- plain and is significant as indicating the former extent of the formation and, by ‘inference of the St. Louis limestone as well, over the crest of the Jessamine dome. A sandstone dike cutting the Lexington limestone in the Kentucky River vicinity is interpreted as Irvine sand preserved in an earthquake fissure resulting from renewed slipping during uplift of the Lexington peneplain. OBSERVATIONS ON SUBSURFACE STRATIGRAPHY IN WESTERN KENTUCKY. Louise Barton Freeman, Ky. Geol. Survey. The study of well cuttings in the search for horizontally persistent features to be used in correlation, whether micro- fossils, heavy minerals or peculiar lithologic succession, is now well underway. There are on file with the Survey only a few well-kept sets of samples, four of which, giving a section from the eastern edge toward the center of the Western Kentucky coal basin, are considered here. The O. V. Brown well in McLean County shows the typical development of the “McClosky.” The Murray Tile Company well, in Breckinridge County, and the Holder well, in Hancock Coun- ty, on the edge of the basin, show a good development of the Salem. The well at Fordsville has a fossiliferous zone at the Salem horizon, but the Brown well has no Salem recognizable as such. The Devonian limestone apparently thickens down dip. At the same horizon in each of these four wells there is a zone par- ticularly rich in pyrite and limonite. In the Brown well there are 43 feet of sandstone resting directly upon the iron-rich bed. The age of this formation and the problem of the value of the iron-rich formation as a horizon marker are Open question at this time. Olt IN EASTERN KENTUCKY. Coleman D. Hunter, Ashland. More than 150 new inside wells were completed in the pools of Estill, Powell and Lee Counties, and a number to be used as intake wells in repressuring operations. The Miller _ Creek extension, the development of a small area in Floyd County and a larger area in Martin County are the only new fields found in Eastern Kentucky dur- ing the year. In the “Big Sinking Pool’, Lee County, development of a small 16 Kentucky Academy of Science extension on Miller’s Creek and drilling some 50 inside wells brought an in- crease in production of over 60,000 barrels. Repressuring operations in the Weir Sand fields of Magoffin, Johnson and Lawrence Counties, and completion of 20 new wells in the Maxon Sand and the Big Lime (Mississippian) in Martin and Floyd Counties produced an increase from Eastern Kentucky of 100,000 barrels over production in 1936. In Martin County, 14 new wells are producing over 200 barrels daily from the Maxon Sand, at a depth of 1100-1400 feet. The structural condition is that of a buried outlier on the old pre-Pennsylvanian land surface. ACTIVITIES AND NEEDS OF THE GEOLOGICAL SuRVEY. D. J. Jones, State Geologist, and Lucien Beckner. During the last four years the Geological Survey has been a division of the Department of Mines and Minerals, under the direction 7f the State Mines Inspector. Appropriations were inadequate for extensive field in- vestigations, and all work had to be done with a minimum otf expense. The first project was the establishment of a repressuring laboratory to aid im the secondary recovery of oil. While the investigations made in this laboratory are not strictly geological, a great deal is being learned about the conditions existing in the oil reservoirs. A great part of the geological work cf the Survey to date has dealt with surface structure and the study of surface stratigraphy. Now’ a laboratory has been established for the study of subsurface stratigraphy as revealed by cuttings from oil and gas wells in the State. A series of pre- liminary bulletins will be published as the work progresses. The Survey has prepared sets of forty rocks and minerals commen to Ken- tucky for use in high schools and junior high schcols. An accompanying bulletin describing each specimen, may serve as an elementary handbook of mineralogy. DEMONSTRATION OF ELECTRICAL RESISTIVITY APPARATUS. R. E. Stouder, Louis- ville. The function of the Gish-Rooney Apparatus is to measure the electrical resistivity of the earth. Four stakes are set in the ground in a straight line and equally distant, with a fifth set between the two center stakes. The commutator reverses the current going into the ground stakes periodically, and reverses the potential connections in exact synchronism. Current is allowed to flow to the outermost stakes and is read “I’ on the ammeter. The voltage “E” is read on the potentiometer. It has been shown that the stake spacing “a” is equivalent to the depth at which measurements are taken; also that the resistance “r” = 27aE /1, After a sufficient number of readings have been taken with increased stake spacings, tha ratios of 2raE/I. are plotted graphically, indicating resistivity against electrode spacing. It has been found by actual work that a new layer will show its presence by a different resistivity when the distance between the stakes is nearly equal to the depth of the new layer. Individual curves in local areas are not similar. The electrical resistivity of the earth’s crust is largely controlled by: (1) the percentage of disseminated moisture, (2) chemical pro- perties and ionization factors of the soluble salts, and (3) the nature of the rock material itself. Twenty-Fifth Annual Meeting 17 MATHEMATICAL ASSOCIATION OF AMERICA Kentucky Chapter D. E. SoutH, Chm. A. R. FEN, Sec. ON REPRESENTATION OF BINARY QUADRATIC Forms. N. B. Allison, Kentucky Wesleyan. MAXIMUM PRINCIPLE FOR ELLIPTIC DIFFERENTIAL EQuATIONS. Fritz John, Univ. of Kentucky. CALCULUS IN BIOLOGY; HIsTorY IN SURVEYING. Two illustrations. W. R. Hut- cherson, Berea College. CONFIGURATION OF DOUBLE POINIS OF CUBICS OF A PENCII.. Sallie Pence, Univ. of Kentucky. A ROLE FOR MATHEMATICS IN THE SCIENCES. W. I. Moore, Univ. of Louisville. SUMMATION OF DIVERGENT SERIES. Mrs. A. 5. Howard Univ. of Ky. THE PLACE OF ASTRONOMY IN THE TRAINING OF HIGH-SCHOOL ‘TEACHERS. Wallace Smith, New River State College. DIVISION OF PHILOSOPHY AND PSYCHOLOGY Kentucky Psychological Association ELLIS FREEMAN, Chmn. L. M. Baker, Sec. Tue DyNAmics oF Instinct. Edward A. Newbury, Univ. of Kentucky. In this analysis of instinct any behavior is considered a resultant of hereditary and en- vironmental factors. Significant features of the problem so considered are in- dicated: instincts as a phase or process in a larger unit of behavior, or as a special characteristic; practice regarded as an error of observation, and the distinction between a habit and an instinct; dynamic variables in both habit and instinct, including not only hereditary and environmental factors, when parts may be separated as instinctive in a whole which is habitual; ease of train- ing and other ways of measuring and controlling the variables. An example of the dynamic approach is provided in observations on the development of swim- ming in rats, normal animals which learn quickly to swim efficiently, abnormal animals which cannot swim as result of a distrubance in the dynamics of the bodily organization. Various factors playing a role in the development of swim- ming are indicated. THE RELATIONSHIP BETWEEN SURFACE TEMPERATURE AND SOCIAL TRAITS IN YouNG CHILDREN. Thelma A. Brown, Univ. of Kentucky. The purpose was to determine whether surface temperature, bore any relationship to social traits in pre-school children. The first part of this study was concerned with the ac- curacy of the instrument and the emotional receptivity of small children to its use. In the second part three nursery-school girls were given readings on a num. 18 Kentucky Academy of Science ber of surface points twenty-one times consecutively for two days, for the pur- pose of checking reliability of various factors. The third part was a study of the relationship between surface temperature and social tendencies for six nur- sery school girls. LocicaL PosttivismM. Ralph A. Gardner, Univ. of Kentucky. This paper is devoted to presenting clearly and precisely the fundamental doctrines of Logical Positivism as they bear upon the relation of philosophy to science. Logical Positivism is concerned primarily with disestablishing metaphysics and with providing a secure foundation for science. Metaphysical systems are founded cn the rationalistic assumption that a transcendent knowledge, which goes beyond empiral verification, is possible. Logical Postivism holds it self-contradictory that the validity of synthetic propositions may be established on rational grounds. All such propositions must have a strictly empirical reference, else they are literally meaningless; and any empirical reference can be demonstrated by logical analysis. All synthetic propositions are scientific hy- potheses; their truth or falsity is a problem for empirical science, not for phil- osophy. In Carnap’s words “The only proper task of Philosophy is Logical Analysis.” A Stupy IN PResByAcusiA. Noble H. Kelley, Univ. of Louisville. This study attempts to answer three problems: First, what loss in auditory acuity, indep- endent of special pathological conditions, accompanies increasing ager The in- vestigation on this problem was concerned with the extent and nature of pres- byacusia in persons ranging from 50 years of age upward. Second, how does such auditory loss affect what one hears in a musical tone? This study involved a comparison of normal and presbyacusic ears. Third, how does such a loss affect our perception of speech sounds? This phase of the investigation attempt- ed to study the effect of presbyacusia on our recognition of vowel and conson- ant sounds. Eight normal observers and twelve subjects with prebyacusia were studied and comparisons made. THe Mopes oF Srmitariry. John Kuiper, Univ. of Kentucky. This paper contains a logical analysis of likeness or similarity. On the basis of the work of Russel, Moore, Langford and others, distinctions are presented between (1) partial and complete similarity of particulars; (2) class-similarity; and (3) re- lation similarity. The question is considered whether the idea of correlation which is basic to (2) and (3) can be utilized in defining (1); or is similarity logically simpler than correlation? THE MARXIAN METHOD. Walter G. Muelder, Berea College. Until very recent years professional philosophers have ignored the philosophy,of Karl Marx and bis direct influence has been almost exclusively with practical, political. and radical groups. This was due in part to the conflict between the basic values of the Marxian philosophy as over against the dominant philosophy of western Europe and Amercia, but it was also due to the decline of Hegelian dialectics out of which Marx’s method evolved. The reasons for the decline in Hegelian Twenty-Fifth Annual Meeting 19 dialectics were: a) the breakup of Hegelian thought into a 1ight-wing and a left-wing; b) the rise of experimental scientific thought which competed with idealistic logic; c) the evolutionary philosophy of the Darwinian popularizers; and d) the non-academic implications of the dialectical method. The general characteristics of dialectical thought in Hegel and Marx are the concrete unity of theory and practice, the significance and reality of wholes, the rejection of mechanistic metaphysics, the historical nature of all social laws, the process of development as thru thesis, antithesis, and synthesis, and the creative activity of thought and consciousness. The Marixan method is distin- guished chiefly from the Hegelian in that Marx is humanistic, revolutionary, realistic in his epistemology. and prospective rather than religious, conservative, idealistic, and retrospective. COLLEGE EDUCATION WITHOUT HIGH SCHOOL GRADUATION—AN EXPERIMENT IN ARTICULATION. Lily Detchens, Univ. of Lousiville. FREUDIANISM AND MODERN PsycHoLocy. Milton B. Jensen, Louisville. AMERICAN ASSOCIATION OF PHYSICS TEACHERS Kentucky Section A. D. HuMMEL, Chm. B. P. RAMseEy, Sec. THe HEAvy ELEcTRON. K. Montgomery. PERMEABILITY OF PoRous MeEp1A. O. T. Koppius. AMPLITUDE AND FREQUENCY MODULATION OF RADIO WaAvES. Louis R. Prince. FIELD STRENGTH MEASUREMENTS AT ULTRA-HIGH FREQUENCIES. Wellington Cochran. UNIPOLAR INDUCTION. F. W. Warburton. KENTUCKY JUNIOR ACADEMY OF SCIENCE Fifth Annual Meeting Eastern Teachers College, Richmond, Ky., April 23. 1938 Over six hundred members representing the 31 Science clubs attended. Pro- fessor W. S. Webb, University of Kentucky, gave the guest address “Prehistoric Life in Kentucky.” Clubs and individuals receiving “Superior” rating and prizes are as follows: I. Best Exhibit: GlassmACe—aBelleviene eri tras tates sinlel ait. ed oe setecenore me prize — $5.00 ClassmBeeMirksvillebon carn on eect te ie ober ition otauei caso oe “i 5.00 II. Best Contribution to Bulletin: @lasse Ane ia EHEC ete SOW his. haptics att Ie chee rete as ae ieee hae 5.00 Glasser ran CES ns SATINULEISS tar yoyy secs es ene es echinacea cee Pin Ill. Best Discussion: CGS JAN. = Ilene? (Chis nmin ey WEN. oss on bo decasaomoodgcosesdous Pin (CMSs. IBS == leaubil |@es RBI —_ oes Bio om ono ae aeOb ode puss oer Pin 20 Kentucky Academy of Science IV. Best Program for Year: Campy Dicks. Robinsons jae sc obi me Seer ieee oe eee Magazine V. New Clubs Organized: welllence, ayavel . Unsknillle, Salm bool 2 etn ado bhapdecacuuseoee: $2.50 VI. Largest Percentage of Pins Owned: ACh Oa GE! iy Nera ape seven a cpey Serer Peasy seal ciedstee wealiy Pellets 2 eh these Magazine IIV. Largest Percentage of Membership Present: Bryan Station, Buena Vista. Danville, White Hall — Each receives a Magazine Clubs recognized for outstanding work: Bellevue, Harrodsburg, Kirksville. The prizes were made possible by cooperation of, Richmond Chamber of Commerce Kentucky Junior Academy of Science Kentucky Academy of Science Officers elected for 1938-39 are, ; TPR ERG IETa Glas a aeeecig SR Pac nat oe ke ey Ea tet oy Denton Russell, Somerset Wii CeSMeSTCIEMIt mis sia eek cess Coe a vos te oe eae ee John Dawson, Bryan Station DECHELANY) oe stomatal Behavior in Field Tobacco iy Stephen Diachum ....- Pe siete Mere ik * DNS Mae ES ae The Reaction Between Alkyl Sulfates and Furoic Acid C. C. Vernon, E. F. Struss, H. H. Ruwe ....--. ines Mase e Mathematical Puzzle as a Stimulus to Mathematical Work Walters be Garvie iy iui ens see a cre nel: a oa ee ye Determination of Inorganic Phosphorus in Corn Grain ag Tcl Ra) Hodder ee pa ' sume of Studies on Beat Tones, Combinational Tones and Sidebands yan Plecbects, kaa lene soya inert U8 We tl eitee dis ua hal ena reason n the Four-Dimensional Mechanism of Knowing | Tester Sap Oannonis Geis eet eo tan hole ee ae OK orecology of Kentucky. I. Claytonis, Mertensia ae and Jeffersonia Piarvey: Ba Povelly 2 s6 20 ie oN not a ies A Study of the Nesting Birds of a Seventy Acre Area in : rae Rowan County, Kentucky (abstract) ean Re WE BAL DOLE) ei, eet an at eae dae aes 48:00) i ——_— Application for entry as second class matter is pending - EDITORIAL STAFF | Charles: Tire) 2000. 0). Murray, Koy _... Editor-in-C Seas Barkenbus)))). 530. Lexington, Kentucky .......... israel aust seat Morehead, Kentucky ............ ] Leta). SD ainwalle item bac kingi a) on sae OW. R.-Hutcherson ....... Berea, Kentucky ..... erie Math ohn Kuiper, 6. os) REA Lexington, Ky. .... Psychology & Philo HH. Bisovelle 2300-3 70 2) ouisville, Kentucky “i 300% SY ae a eke G: McParlan \ 0. 25" ou Lexington, (Mentuckys |) 4 aye HM _ Manuscripts. The Transactions must be limited to the proceed the annual meetings of the Kentucky Academy of Science and to or manuscripts pertaining to science. Under present financial limitations article must be limited to approximately five pages of the Transa t material may be submitted to the Associate Editor of the subject covere to the Editor in Chief. Exrra-Cost Features. ‘The extra cost of special features such as graphs, tables, etc. above the text-run price of $2.65 per page must be bors by the contributor. The Editorial Staff will advise contributors concernin: _ the extra cost of special features upon receipt of manuscript. 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Remittances and correspondence coné _ing subscriptions, extra costs, and other financial matters except repr should be addressed to Chas. Hire, ay State Teachers cores M Kentucky. a 4 Twenty-Seventh Annual Meeting 17 i ; ‘ SPECTROGRAPHIC IDENTIFICATION OF MINOR ELEMENTS IN HAY AND GRAIN MIXTURES 4 W. S. Hopexiss anp B. J. ERRINGTON Agricultural Experiment Station : University of Kentucky ; Studies of the presence of the “trace” elements in plant or animal tissue ; have been greatly handicapped in the past due to the lack of appropriate _ procedures of chemical analysis, sensitive enough to detect the presence of : _ such elements in a relatively small portion of the tissue. Different quantities of inorganic elements are taken from the soil by different plants. This is as true of the trace elements as it is of those which are distributed more abundantly. However, the presence of un- usually large quantities of certain elements in plant or animal tissue gives rise to a question as to whether these elements are present for a definite purpose and therfore may be considered essential for animal or plant growth. The importance of the presence of small quantities of compounds of metallic elements in plant material has been emphasized especially with reference to their requirements in proper growth of the plant. Similarly, animal feeding trials have shown that a number of the less common elements are essential for an animal diet and which, along with the proper organic nutrients and vitamins, are necessary for proper development in the animal. Not all, however, of the trace elements found in soils and plants are essential or even beneficial to animal or plant life. Two are known to be definitely harmful and others may be under certain conditions. Little is br known, as yet, of the role that the greater proportion of the trace elements % take in plant or animal life. The samples selected for comparison in this work were three hay and 4 three grain mixtures which have been used as feed for livestock in this vicin- ity. As is the case with many such feeding mixtures, the main source of : materials was from out of the state. The grains and hays are designated be _as A, B and C for convenience in comparison. _ The grain mixtures were similar in composition and based on varying _ proportions of oats, corn, wheat bran and salts. Grain C, however, had no f salts added as was the case of the other two mixtures. The hay mixture C was made up of lespedeza and timothy and was considered from its appearance poor in nutritive value. Hay A was termed an 80% clover hay while B was a mixture of 60% clover and alfalfa. S| Re Ser i ae, Ow Ie ee Vee bee S pectrographic Identification of Elements. When a substance is heated to incandescence or excited electrically, each of the elements present emits light of wavelengths which are characteristic ‘ of that element. This light is split by a quartz or glass prism into a “spectrum and recorded as lines on a photographic plate. The elements present may be identified by comparing the position of these lines with 18 The Kentucky Academy of Science those of the known lines of different elements. In general, the iron arc spectrum is used for identification and comparison of the wave lengths. The intensity of a spectral line depends upon many factors besides the abundance of the element in the source. However, if for qualitative esti- mation these factors are neglected, the concentration of the element may be regarded as a function of the logarithm of the intensity or blackening of the line on the photographic plate. The instrument used in this study was a large, Littrow type spectrograph utilizing a quartz prism with a dispersion of about 35 inches over a range of 2000 to 6500 Angstroms. ‘There are variations in the sensitivity of lines appearing on the spectrographic plant for each element, a line which is visible at one concentration may not be present at another. Each element has at least one line which is more intense than the others and is the last to dis- appear on reduction of the total amount of element present. These lines are used as a means of determining the limit of sensitivity of the procedure. Since there is a variation in the sensitivity of lines for different elements, the minimum concentration at which a line disappears is not the same for each element. The limit of sensitivity of lines used in identification of the element was approximately as follows: 1) Oy =) Gi Ning Al Crab: 10) = 100y > — Co, Nite Moje Va) Tay Sni by bi. Basser £00 S=1000y = Je Sbs iGds Ane shy llr ips we Estimations of the concentration of metals were made by a modification of the comparison method used by Nitchie (1) and Rusoff, Rodgers and Gaddum (2). Standard solutions containing known percentages of 26 elements at four dilutions were arced in positions adjoining those of un- known samples on the photographic plate. Visual comparison of the line densities gave an indication of the approximate concentration of the element in the sample. A small portion of the ashed plant material, varying between 1 and 50 mgm., depending upon the sensitivity of the lines of the element determined, was placed in a crated carbon electrode and heated in a 240 volt arc with a current of 11 amperes until completely volatilized. The data are presented in a “range” form and should be read as 1 to 10 mgm. per kilogram indicating that the concentration of the element lies somewhere between one and ten mgm. per kilogram of the moisture free material. The analyses indicate that the hays and grains were similar with respect to the elements identified. There appears to be no definite relationship between the elements found in the hays or in the grains. The hays were found to be high in aluminum, barium, strontium and boron. The con- centration of lead, lithium, zinc and cobalt was found to be greater in the grains than the hay mixtures. Cobalt was found to be present in the grains but could not be positively identified as being present in the hays in a 50 mgm. sample of the ash. Similar concentrations of the other elements were found in both the hays and the grain mixtures. Twenty-Seventh Annual Meeting 19 Ten of the twenty-six elements were not detected in the plant samples. Spectrographic Estimation of Trace Elements in Hay and Grain Mixtures. (Mgm/kilogram m.f. material) 100-300} 100-300} 200-400 5-20 1-10 1-10 ge .1-1 re <.l <1] <1 ? ? ? .1-10 .1-10 .1-10 1-1 1-1 1-1 <1 1-1 <.l 1-10 1-10 1-10 2-20 5-30 2-20 Not detected—Bi, Cd, W, Rb, Cs, Tl, Zr, Be, Th, Sb. No attempt was made to concentrate solutions of the ash. It is probable that elements which were not detected in the grain and hay mixtures could be found to be present after concentration or chemical separation. The term “‘trace” used in the table signifies that a line was faintly visible but no estimation could be made of its approximate concentration. All of the less common elements identified in the plant material will eventually be studied from a quantitative standpoint with reference to their presence in plant and animal tissue. Summary: Three hays and three grain mixtures used as feed for livestock in Ken- tucky were analyzed by spectrographic means for 26 trace elements. 20 The Kentucky Academy of Science The following elements were detected in all of the samples of hay and grains: copper, manganese, zinc, barium, strontium, lithium, tin, molybdenum, aluminum, lead, chromium, nickel, cobalt, vanadium, titan- ium and boron. Elements not detected were: bismuth, cadmium, tungsten, rubidium, cesium, thallium, thorium, zirconium, beryllium and antimony. The hays appeared to be higher in aluminum, barium, strontium and boron than the grains. The grains appeared to be higher in lead, lithium, zinc and cobalt. (1) Nitchie, C. C. Ind. and Eng. Chem. Anal. Ed. 11; 1; 1929. (2) Rusoff, LL, Redgers, Il. and Sta LW Ase Research 55; TINO 37 STOMATAL BEHAVIOR IN FIELD TOBACCO* STEPHEN DiacHUM Agricultural Experiment Station University of Kentucky A close correlation between stomatal condition and infection of tobacco leaves by Bacterium tabacum, the wildfire organism, was reported in a previous paper from this laboratory (1). Stomata are pores or openings in the epidermis of leaves. Their size is regulated by the expansion and contraction of specialized guard cells, and they may therefore at a given time be either open or closed. Infection occurs only when they are open. Thus before we can have a thorough understanding of the mechanism of leaf infection by bacteria, it is necessary to know at what times or under what conditions stomata are open or closed. METHODS Stomatal width can be determined directly by microscopic measurement, but this is a slow, tedious method. We have developed an indirect method which is more convenient and more rapid. If a stream of water is forced against the lower surface of a tobacco leaf, water is driven through the stomata into the leaf and floods the tissue, producing visible water-soaking. The rate at which water-soaking occurs depends on the degree of opening of the stomata; water-soaking occurs very rapidly when stomata are wide open and very slowly or not at all when they are closed or narrow (2). In the studies which are reported here, the rate of water-soaking was determined by directing a stream of water from a 10-c.c. hypodermic syringe against the lower leaf surface. The rate at which water soaking occurred is expressed as very fast (VF), fast (F), slow (S), very slow (VS), or not at all (O). In conjunction with this indirect method, stomata were measured microscopically. For stomatal measurement a small piece of lower epidermis was stripped and plunged into absolute alcohol, and during this microscopic examination the epidermis was mounted in Twenty-Seventh Annual Meeting 21 absolute alcohol. The average width (in microns) of 20 measured stomata, was found to be a sufficiently accurate index of the degree of stomatal opening. All observations were made on field-grown Burley tobacco. OBSERVATIONS In general stomata were found to be open during the day, and closed or nearly closed at night. In table 1 are presented notes on three days’ observ- ations on stomatal width and the rate at which leaves could be water-soaked. These data are typical. TABLE 1. Effect of time of day on stomatal opening of field-grown tobacco. July 10 & 11, 1939] July 18 & 19, 1939] July 19 & 20, 1939 Time | Width of | Rate of | Width of | Rate of | Width of | Rate of of stomata water- stomata water- stomata water- Day soaking soaking soaking 3:30 microns microns microns p-m 3.8 VF 7 p.m 1.4 VS 1.0 0 0.6 0 8 p.m 1.0 0 0.5 0 10 p.m 0.9 0 0.8 0 1 am 0.9 0 3:30 a.m 0.6 0 5 a.m 1.9 VS 6 a.m ; 4.0 F 7 a.m adh VE 8 a.m 4.7 Wola 4.6 VE 4.1 VE 10 a.m 4.7 VFE 4.1 VF Stomata were not, however, always open during the day. It was observed several times that when light intensity dropped below 100 foot candles (dur- ing very dark storms) stomata were closed or nearly closed. Several other times stomata were found open when light intensity was as low as 300 ft. 22 The Kentucky Academy of Science candles. ‘The critical point at which they begin to close is not known; nor do we know how rapidly they close after light is reduced—as, for example, on the approach of storms. The regulatory action of light is also demonstrated by the fact that stomata on shaded leaves were usually found to be closed, while those on unshaded leaves of the same plant were open. A few cases are presented in table 2. The degree of shading and time of shading required to close stomata is not known. TABLE 2. Effect of shade on stomatal width, and rate of water-soaking Date Shaded leaf Unshaded leaf Stomatal Rate of Stomatal Rate of width water- width water- soaking soaking Microns Microns 7ANG=39 .m. 0.9 3.0 7-17-39 7-18-39 7-31-39 Furthermore, frequently at midday or in the afternoon when the sun was bright and light intensity was as high as 7000-8000 foot candles, stomata were closed on some of the upper leaves. Usually at such times the leaves were being blown so that the lower leaf surface was exposed to the sun. It is not known whether such stomatal closure is due to incipient wilting of the leaf because of the drying action of the wind and sun, to excessive sunlight on the lower epidermal cells, to low humidity, or to some other factor. Stomata on wilting leaves were often closed or partly closed; tho some- times they were open on leaves that were quite noticeably wilted along the tip and edges. Wilting was produced experimentally several times by cut- ting the midrib at about the center of the leaf; in such instances stomata closed on the wilted tip half and remained open on the basal half of the leaf. CONCLUSION It is to be concluded that although stomata of tobacco leaves tend to be open during the day and closed at night, there are several modifying factors which may upset this tendency. A close correlation exists between stomatal width and rate at which leaves can be water-soaked: when stomata Twenty-Seventh Annual Meeting 2B are open, leaves can be water-soaked very rapidly; when stomata are closed, leaves can be water-soaked very slowly or not at all. * The investigation reported in this paper is in connection with a project of the Kentucky Agricultural Experiment Station and _ is published by permission of the Director. (1) Diachum, S. Phytopathology 30; 268-272; 1940. (2) Diachum, S. and Valleau, W. D. Am. J. of Botany 26; 347-351; 1939. THE REACTION BETWEEN ALKYL SULFATES AND FUROIC ACID C. C. VeRNon, E. F. Struss, anD H. H. RuwE- Department of Chemistry, University of Louisville In connection with some syntheses attempted a few years ago in this laboratory some data were collected concerning the preparation of the methyl and ethyl esters of furoic acid. There seems to be little in the literature concerning the esterification of this acid with methyl and ethyl sulfates, although this is a common method for introducing alkyl groups. Esters of furoic acid have commonly been prepared by passing hydrogen chloride into alcohol solutions of furoic acid (1) (2). While furoic acid is probably the most stable of all the simpler furan derivatives, the stability of the furan ring under such conditions is rather surprising (3). Recently the methyl and ethyl esters of furoic acid have been made by adding furoic acid to a previously prepared solution of sulfuric acid in alcohol, and then heating the mixture for several hours (4). In view of the instability of furan compounds in the presence of mineral acids (3), and considering the rather large amount of acid used (approximately 15 mols in 43 mols of alcohol), yields of 60-65 per cent were surprisingly high. A logical explanation seemed to be that the alkyl sulfate was first formed and that this reacted with the acid to form the ester. If this be the case the use of known alkyl sulfates should at least duplicate the yields and might prove advantageous in other ways. Naturally, we hoped to increase the yields. Lewis and co-workers (5) have reported certain optimum conditions for the alkylation of phenol with alkyl sulfates. They found the best pro- portions for their work to be, two mols phenol, three mols sodium hydroxide, one and six tenths mols water. Such a mixture was heated on a water bath, and then one mol dialkyl sulfate was slowly added, the whole being heated for three hours longer. At the end of this time three hundred cubic centi- meters of one per cent sodium hydroxide was added, and the product cooled and extracted with ether. According to Lewis (5), dimethyl sulfate commonly reacts in aqueous solution to give up only one of its methyl groups to the reaction, but if little or no water was present he found both methyl groups entered into the The Kentucky Academy of Science TABLE I—Furoic acid with dimethyl and diethyl sulfates Run ey 2p ee Sa SB Ain oS Oia tS Ma ee eee 0.1 0.042 0.1 0.45 1 0.1 0.021 0.15 0.45 3 0.1 0.042 0.15 0.45 1 0.1 0.042 0.15 0.45 3 0.1 0.042 0.2 0.45 1 0.1 0.042 0.2 0.45 1 0.1 0.042 0.2 0.45 3 0.1 0.042 0.2 0.45 3 0.1 0.1 0.1 0.33 4 Oylial Ont 0.1 0.22 4 0.1 0.1 0.1 0.16. 4 0.1 0.1 02 0.22 4 0.1 0.1 0.2 0.00 4 0.1 0.1 0.2 0.00 4 0.1 0.1 0.2 0.33 4 0.1 0.1 0.2 0.22 4 Ont. 0.1 0.2 0.00 4 0.1 0.1 0.2 0.00 4 0.1 0.1 0:2 0.00 4 0.1 0.1 0.1 OL00L 5 4 0.1 0.1 0.3 0.33 4 0.1 0.1 0.3 0.00 4 ~NasCO3; 0.1 0.1 0.1 0.00 4 a. toluene as solvent. Twenty-Seventh Annual Meeting 25 reaction. Diethyl sulfate normally gives up both of its ethyl groupings. In this study it was proposed to determine to what extent those conditions applied in this reaction, as well as to attempt to determine the mechanism of the esterification reaction previously mentioned (4). EXPERIMENTAL The furoic acid was prepared from furfural by the Cannizzaro reaction (6). Recrystallization from benzene was found to give a product superior to that secured when recrystallization was from water. Recrystallization of turoic acid in this manner is now common practice in our laboratories and several others have recently adopted it. Technical dimethyl sulfate was used in the first eight runs, but it was analysed and the mol fractions caluclated for Table 1 on the basis of this analysis. The alkyl sulfates used subsequently were purified according to standard methods (7) (8). The most satisfactory method for our purpose was that of Lynn and Shoemaker (8) which involved merely washing once with sodium carbonate solution, then several times with cold water, and finally drying over anhydrous sodium sulfate. The procedure followed in running the experiments was essentially that used by Lewis (5). A typical run was carried out in the following manner. Furoic acid, potassium hydroxide and water, in the quantities indicated in Table 1 were mixed in the reaction flask and then stirred and heated for one hour over a water bath. The alkyl sulfate was then slowly added, while the warm reaction was thoroughly stirred. The whole mixture was then stirred and heated over the water bath for the time indicated in Table 1. When cooled, it was extracted several times with ether, and this ether extract distilled to secure the ether. In those cases where Table 1 indicates that there was no water in the reaction mixture, the furoic acid and potassium hydroxide were mixed in water and this evaporated to dryness over a water bath. This mixture was then transferred to the reaction flask and the alkyl sulfate added as before. DISCUSSIONS In the first eight experiments with dimethyl sulfate, the time of heating and the amount of alkali were the variables. In the second experiment only 0.021 mols of dimethyl sulfate was used unintentionally, and the heating prolonged to three hours. An unexpectedly good yield resulted, which we were unable to duplicate, or explain. In this group of experiments, seven and eight gave the best yields, which were repeatedly duplicated. In the next group of experiments, the dimethyl sulfate used was purified, as previously mentioned, and the amount increased to 0.1 mol. The heating period was increased to four hours, which subsequent experiments not shown in Table 1 proved to be the maximum from which advantage could be derived. In three of these experiments, 9, 10, and 11, 0.1 mol of alkali was © used, and the amount of water decreased. Apparently 0.22 mol of water 26 The Kentucky Academy of Science was best, which was apparently substantiated in experiment 12, where the alkali was again increased to 0.2 mol. Experiments 13 and 14 in which no water was used gave substantially lower yields. Repetition of experiment 12 gave yields of 50 to 51%, and confirmed our belief that these were the optimum conditions. Experiments 15 to 20 were with diethyl sulfate, and based on experience with the dimethyl sulfate Those chosen for Table 1 represent extremes, which for the most part were unexplainable. However the conditions and yields shown in 18 and 19 were duplicated often enough to convince us they were the best. There was no explanation for the low yield in 17. In experiments 21 and 22, 0.3 mol of. alkali was used, which seemed de- cidedly disadvantageous with each of the dialkyl sulfates tried. It was strange that the amount of water should have had so great an effect, and that this effect should vary in the two cases studied. No logical explanation for this can be offered at the present time. In most reactions there was slight charring indicating some decomposition. This was hardly to be expected, since furoic acid was fairly stable in the presence of alkali, and there should have been no free sulfuric acid in those reactions where 2 mols of alkali were used. In the last experiment, a method studied by Haworth and Lapworth was tried (9) (10). In this method sodium carbonate was used, as the alkali, and anhydrous toluene served as a solvent. There appeared to be no re- action under these conditions. CONCLUSION Furoic acid has been esterified by the use of dimethyl and diethyl sulfates. The best yields attained with dimethyl sulfate were secured when 0.1 mol acid, 0.1 mol sulfate, 0.2 mol alkali, and 0.2 mol water were refluxed for 4 hours. With the diethyl sulfate, the same proportions were best, except that the best yields were secured when no water at all was used. Other concen- trations of reagents were investigated but gave poorer yields. (1) Gennari. Gazz. chim. ital. 24; 253; 1894. (2) Zanetti and Beckman. J. Am. Chem. Soc. 48; 1067; 1926. (3) Gilman and Vernon. J. Am. Chem. Soc. 46; 2576; 1924. (4) Private communication. (5) Lewis, Shaffer, Trieschmann and Cogan. Ind. Eng. Chem. 22; 34; 1930. (6) Gilman. Organic Syntheses. John Wiley and Sons. VI; pp.44-46; 1926. (7) Guyot and Simon. Compt. rend. 169; 795; 1919. Chem. Ab. 14, 4.035) 59s 920» (8) Lynn and Shoemaker. J. Am. Chem. Soc. 46; 999; 1924. (9) Haworth and Lapworth. J. Chem. Soc. 123; 2986; 1923. (10) Hodgson and Nixon. J. Chem. Soc. L930: Twenty-Seventh Annual Meeting oy, THE MATHEMATICAL PUZZLE AS A STIMULUS TO MATHEMATICAL WORK (condensed from one hour address) WaLTeR B. CARVER President of Mathematical Association of America — The problems that appeared in the algebra and geometry texts of forty years ago have been criticized in our day as being too artificial; and we are told that if interest in mathematics in the schools is to be maintained our problems must be more practical and more closely related to business, industry, and everyday life. While generally sound, this point of view over- looks one important bit of educational psychology; namely, the wide-spread interest in puzzles. Our algebra used to ask us “at what time between three and four o’clock are the hands of the clock together,” and the modern criticism is, “Who cares?” But the puzzle character of this problem made a certain appeal, and the interest was in no way diminished by the fact that the answer, when obtained, was of no particular use to any one. It is somewhat amusing that as our modern text-books present problems that are more useful but not always interesting, we must look to puzzle columns in the Sunday newspapers and Esquire magazine to furnish us with some ex- cellent mathematical material of a highly entertaining character. Many a boy or girl who is bored to death with formal drill and useful applications in algebra and geometry will show keen interest in a clever, though entirely useless, puzzle. And in solving the useless puzzle he may become familiar with the very same mathematical processes which are needed to solve the more useful problems which arise in business and industry. The five following puzzle problems illustrate the type of material. The mathematics involved ranges from the very trivial elementary algebra in the first to fundamental calculus notions involved in the fifth. 1. MATHEMATICAL MIND-READING. Ask any one to choose any number less than 20, and then do the following arithmetic: add 3 to the chosen number; multiply the result by 2; subtract this result from 77; multiply the result by 5; to this result add 10 times the original number. You can then tell him his final result. The procedure is obviously subject to many kinds of variations. 2. COMPUTATION OF ANNIE’S SURNAME. Three women, Mrs. Brown, Mrs. Jones, and Mrs. Smith, each with her daughter, bought material at a store, each of the six persons paying as many cents per yard as the number of yards she bought. Each mother spent $1.75 more than her daughter. Mrs. Brown paid almost ten times as much per yard as Mary, and Bessie spent 31 cents less than Mrs. Smith. What was Annie’s last name? 3. KIDNAPPERS FOILED BY ALGEBRA. Kidnappers asked a ransom of only $200 for the return of a banker, but specified the following difficult terms of payment: the money was to be paid in 11 payments 28 The Kentucky Academy of Science on 11 successive days; any amount might be paid the first day, and any amount the second day; the third payment must be the sum of the first two; the fourth the sum of the second and third; and each later pay- ment the sum of the two preceding payments. The total must be exactly $200, neither more nor less. Fortunately the banker’s son was good at algebra and was able to make the payments as required and thus secure his father’s release. 4. CHICAGO A HALF GAME AHEAD—BUT ACTUALLY BEHIND. At a certain time during a close National League pennant race Chicago has won 54 games and Pittsburgh has played 88 games. Pittsburgh’s percentage is higher than that of Chicago, but Chicago is what the sports writers call ‘‘a half game ahead” of Pittsburgh. How many games has each team won and lost? 5. SCRAMBLED HATS. Eight men had been dining not wisely but too well at a certain London restaurant. They were the last to leave, but no one man was in a condition to identify his own hat. Now consider- ing that they took their hats at random, what are the chances that every man took a hat that did not belong to him? If K, represents the number of ways in which n hats may be distributed among n owners without any man getting his own hat, then K, is the integer nearest to n!/e and e—*/(1-x) = 1 + 3 K,x/i! i=) THE DETERMINATION OF INORGANIC PHOSPHORUS IN CORN GRAIN Jack R. Topp Agricultural Experiment Station University of Kentucky Certain workers in recent years have expressed the belief that an analysis of plant sap or green tissue would serve as a better measure of the availability of the soil phosphorus than an extraction of that soil by chemical solvents. This idea has never been carried over to matured and dried tissue. Dr. Weeks, of the Agronomy Department, Kentucky Experiment Station, suggested that a measurement of the inorganic and organic phosphorus con- tent of such dried materials as corn grain and stover might show whether the increased phosphorus content of this material when grown on plots receiving phosphate fertilizers, was held mainly in the inorganic form and whether the inorganic phosphate content would parallel the availability of the soil phosphorus as determined by chemical or other means. Before any answer to these questions could be found, certain difficulties presented themselves in the determination of the inorganic phosphorus in these ma- terials by the usual methods. After reviewing the literature on the problem of the determination of inorganic phosphorus, especially some of the earlier work, the conclusion Twenty-Seventh Annual Meeting 29 was reached that some of the present suggested methods did not fully take into account certain difficulties that are involved in the determination. While they determined a definite portion of the phosphorus present, it was not necessarily inorganic phosphorus alone they were measuring. In order to make a study of these difficulties somewhat easier, they have been grouped under five headings. As is usual with many such classifi- cations, some of these difficulties were not confined strictly to one group- ing, but more or less overlap into other groups. However, in general they were: (1) The extractant (2) The time of extraction (3) The effect of organic phosphorus compounds (4) The effect of non-phosphorus compounds, and (5) The nature of the plant material. (1) The extractants: The extraction usually has been in an acid medium, either sulfuric, hydrochloric or acetic with or without added compounds as alcohol or molybdate. In 1912 Collison (2) suggested the use of 0.2% hydrogen chloride in 95% alcohol and shortly after that Heck & Whiting (8) modified the pro- cedure to 1% hydrochloric acid which, on the whole, seems to be a rather insignificant change. Before Collinson’s method was suggested, Forbes (6) had used 0.2% hydrochloric acid, the same as was used in the Heubner and Stadler method for the extraction of phytin phosphorus. Forbes’ method was a modification of that of Hart and Andrews (7) who tried both 0.2% hydrochloric acid, 1% acetic acid, and each of the acids in combination with tannin. Hart and Andrews had found that the amount of inorganic phosphorus extracted varied with the concentration of the acid and varied also with the particular acid, hydrochloric extracting more than acetic, but selected 0.2% hydrochloric acid as being the most satisfactory. Rather (14) in studying the extraction of phytin phosphorus, tried 0.2 % hydrochloric, then 1.2%, and found that 1.2% extracted more than the 0.2 % ; therefore, he concluded 1.2% was better and used it. Emmert (5) extracts crushed green plant tissue with 1% by volume sulfuric acid. Thornton (15), Pierre and Pohlman (12), Chapman (1) and others (3, 9, 11) were some of the more recent workers who worked particularly with green plant tissue or plant sap. They have all used acid-ammonium molybdate, usually in 0.75 normal acid, either hydrochloric or sulfuric. Most of the later workers, at least as expressed by their published results, did not take into account the effect of the kind or concentration of acid, but used an arbitrary amount that was suitable or that had been previously suggested, (2) Time of extraction. Hart and Andrews tried different extraction periods and found that varying amounts of phosphorus were extracted with different lengths of time but that the amounts were not always in 30 The Kentucky Academy of Science proportion to the time. They finally found that a fifteen-minute extrac- tion period, with the extraction flasks being shaken constantly, was the most suitable. Forbes, who used the work of Hart and Andrews as a basis for his studies, came to the conclusion that a fifteen-minute period was not long enough for complete extraction of inorganic phosphorus, so he used a 3-hour extraction period with occasional shaking of the flasks. Some of the more recent writers, working with green plant tissue, such as Thornton, merely shake the material with the extractant, no definite time being given, while Emmert extracts for a period of five minutes before filtering and developing the blue color. (3) Organic phosphorus. A discussion of the effect of the organic phosphorus compounds present would overlap, at least in part, the dis- cussion on the nature of the extractant and the effect of time, for these two factors had considerable bearing on the organic matter present and its effect. Miller, in his book “Plant Physiology,” lists phytin, phospholipids, hexose phosphates, neucleic acid and phosphoproteins as the organic phosphorus compounds in plant tissue. In making a determination for inorganic phosphorus, one would have to be certain that the extractant was of such a nature as to prevent enzyme- splitting of the organic compounds that were present and not in such con- centration as to cause acid-hydrolysis. Hart and Andrews criticized the work of Iwanow, who used 1% acetic acid, and Zaleski, who used both 1% acetic and 0.2% hydrochloric, on the grounds that they failed to take into account the cleavage action on soluble neuclein of the nitric acid in the molybdate solution. They remedied this by adding ammonium nitrate and ammonium molybdate rather than the usal acid molybdate, making a neutral precipitation of the phosphorus. Forbes criticized Hart and Andrews’ results because (a) the time was in- sufficient for complete extraction, (b) the precipitation with neutral ammonium molbydate was difficult, (c) a precipitate that was formed made it difficult to determine if the inorganic phosphorus had been com- pletely precipitated, (d) the solution contained a precipitate difficult to filter, and (e) complete precipitation of inorganic phosphorus in the presence of phytin was impossible. Forbes solved these difficulties by extracting for three hours with 0.2% hydrochloric acid, filtering and taking an aliquot, adding magnesia mixture to the aliquot, and then making it slightly alkaline with ammonia, thus precipitating the inorganic phosphorus. The precipitate was filtered, washed with alcohol, and dissolved in acid alcohol. An aliquot was then taken, dried, and the phosphorus determined. Collison modified the method of Forbes by using acid-alcohol as the extractant which he said did not dissolve either phytates or neucleic acid but did dissolve the phosphates. _ Enzymes could not act in this alcoholic medium and the acid concentration was not too strong. DeTurk (4) did not attempt to state definitely what type of phosphate he was extracting but instead divided the phosphorus into solubility groups Twenty-Seventh Annual Meeting 31 as (a) alcohol soluble, (b) acid-alcohol soluble, alkaline insoluble, (c) acid soluble and (d) acid insoluble. Chapman stated that in the method he used, 0.75N HCl in ammonium molybdate, he was not certain that the test distinguished quantitatively between inorganic phosphate and certain forms of organic phosphate. However, if any important relationship was found between the phosphate determined and soil treatment, it was immaterial whether the distinction between inorganic and organic was absolute from a practical point of view. Chapman stated that Plimmer (13) working with the hydrolysis of various phospho-organics found they were not hydrolized under the conditions of extraction as used in his procedure. One advantage more recent workers had over previous workers lies in the use of one of the modifications of the Denig’s method for phosphorus, as Truog and Meyers’ (17). ‘There is no prolonged precipitation in which hydrolysis might take place as in the older A. O. A. C. method for de- termining phosphorus. (4) Non-phosphorus compounds. ‘The only workers who have taken into account non-phosphorus compounds are (a) Collison, who stated that carbohydrates which prevent complete precipitation of phosphate are not soluble in the acid-alcohol extractant which he used and (b) Forbes who added definite amounts of inorganic phosphorus to plant materials be- fore extraction and obtained complete recovery. Some others have carried out this same idea of recovery of added amounts of phosphorus. (5) The material extracted. Without going into the problem of what time during the growing season is the proper time for analysis or what tissue to take, one finds there is considerable variation with season in the per- centages of the different types of phosphorus compounds in plants, as well as total phosphorus. Tottingham, in his “Plant Biochemistry”, presented data to show how the phosphorus composition varied during the growing season in normal and in treated or starved plants. Most of the recent workers used green tissue and Truog and Meyers’ method for determining phosphate phosphorus. There has been some sug- gestion, however, that the method is not satisfacory for dried material, especially dried forage. | In our laboratories the method suggested by Collison was used. An acid-alcohol extraction was made with subsequent ammoniacal precipitation of the inorganic phosphorus. However, either the Truog or Fiske and Sabarrow colorimetric method for the determination of the phosphorus extracted was used, depending on the concentration of phosphorus, instead of the A. O. A. C. gravimetric method. Also an aliquot was taken from the acid-alcoholic extract, evaporated to dryness, digested in perchloric- nitric acid mixture, and the phosphorus determined colorimetrically. After dissolving the alkaline precipitate in acid, considerable difficulty was encountered in obtaining clear solutions or solutions that did not give such off-color tints that they could not be compared with the standard. 32 The Kentucky Academy of Science % Phosphorus acid- alcohol | In- Sample Treatment Time | Total | soluble | organic Effect of Time. Composite A | 25 cc 1% acid alcohol] % hr. | 052 .013 Composite A} 25 cc 1% acid alcohol] 1 hr. .061 014 Composite A| 25 cc 1% acid alcohol} 2 hrs. shy .067 .018 Composite A| 25 cc 1% acid alcohol] 5 hrs. .085 .026 Composite A} 25 cc 1% acid alcohol} 7 hrs. 100 .028 Composite A} 25 cc 1% acid alcohol] 17 hrs. sll 032 Composite A 1% acid alcohol Composite A 1% acid alcohol Effect of Acid Concentration Composite B| 25 cc 1% acid-alc. 9 hrs. Composite B 3% acid-alc. 9 hrs. Composite B 6% acid-alc. 9 hrs. Effect of Extractant Composite C} 25 cc distilled water Composite C| 25 cc 1% HCl Composite C| 25 cc 1% acid-carbon tetrachloride Composite C cc neutral alcohol Composite C ce 1% acid-alcohol Comparison of Total With Acid-Alc. Soluble 309 HG 25 cc 1% acid alc. 9 hrs. .107 .027 309 SG 25 cc 1% acid alc. 9 hrs. 086 .022 310 HG 25 cc 1% acid alc. 9 hrs. 072 .024 310 SG DS ec liJo acid) ale: Oe hrs: 094 .042 311 HG 25 ce) 1%) acid alc. 9 hrs. 072 .022 311 SG 25 cc 1% acid alc. S hts: BA eo a| 317 HG 25 cc 1% acid alc. 9 hrs. 112 317 SG 25 cc 1% acid alc. 9 hrs. 144 318 HG 25 cc 1% acid ale. 9 hrs. 094 318 SG 25 cc 1% acid alc. oy hes: 125 901 HG 25 cc 1% acid alc. 9 hrs. .094 9 hrs. 25 cc 1% acid alc. Twenty-Seventh Annual Meeting 33 A study of the effect of time on the phosphorus extracted showed that there was a sharp rise in the amount extracted up to about 9 hours, and a decrease in the rate from that time on to about 24 hours. Extraction beyond 24 hours showed little or no increase in the amount of phosphorus removed. With an increased acid concentration there was a corresponding increase in the amount of phosphorus dissolved. However, the difference betwen 1% and 3% acid was much greater than the difference between 3% and 6%. Certain other extractants were used also to see how they compared with the acid-alcohol extractions. The phosphorus determined in these cases was that dissolved by the extractant, not the inorganic phosphorus. Water dissolved more than either 1% hydrochloric acid or acid-alcohol; the acid dissolved more than acid-alcohol; neither acid-carbon tetrachloride nor neu- tral alcohol dissolved any phosphorus. A comparison of the total phosphorus with the acid alcohol soluble showed, in general, that the acid-alcohol solution was directly related to the total present. With only limited results, the inorganic phosphorus showed little correlation with the total phosphorus present or soil treament. From an average of the determinations made, it was found that of the total phosphorus present, 10% was in the inorganic form and 30% was acid-alcohol soluble. Thus, in brief, it was found that the determination of inorganic phos- phorus in dried plant material, as corn grain, was not a simple one. Also the results obtained did not appear to have any greater significance than those of total phosphorus. (1) Chapman. Soil Sci. 39; 111 (2) Collison. J. Biol. Chem. 12; 65 (3) Culbert and Hendrin. J. Agr. Res. 35; 185 (4) DeTurk. J. Agr. Res. 46; 121 (5) Emmert. Plant Phys. 5; 413 (6) Forbes, Lelmann, Collison and Whettin. Ohio Exp. Sta. Bul. 215 (7) Hart and Andrews: New York Agr. Exp. Sta. Bul. 238 (8) Heck and Whiting. Soil Sci. 21; 17 (9) McCool. Mich. Agr. Exp. Sta. Quart. Bul. 9; 60 (10) Miller. Plant Physiology (11) Pettinger, J. Agr. Res. 45; 95 (12) Pierre and Pohlman. J. Amer. Soc. Agron. 25; 144 (13) - Plummer. Biochem, J. 7; 72 (14) Rather. J. Amer. Chem. Soc. 39; 2506 (15) Thornton. Ind. Agr. Exp. Sta. Bul. 355 (16) Tottingham. Plant Biochemistry (17) Truog and Meyers. Ind. and Eng. Chem. Anal. Ed. 1; 65 34 The Kentucky Academy of Science RESUME OF STUDIES ON BEAT TONES, COMBINATIONAL TONES AND SIDEBANDS HERBERT HAZEL Department of Science, Ashland Junior College Helmholtz-Koenig Controversy. iNboue tae anidtlle af aie (oah century the German organist, Saree dis- covered that two musical tones of different pitch sounded together loudly and continuously gave rise to a new tone. A little later Tartini, an Italian violinist, also recognized such tones. The frequency of the derived tone observed in each case was equal to the difference between the combining frequencies, a fact which led Chladni, Lagrange, and Young to the view that the new tones were merely quickened beats. As the beats became too rapid for individual appreciation, the effect of a continuous frequency was produced. About a century later Helmholtz confirmed the existence of the tones observed by Sorge and Tartini, and also detected derived tones having frequencies equal to the sums of the frequencies of the combining tones. To the tones found by Sorge and Tartini Helmholtz gave the name “dif- ferential tones” and to those which he discovered he gave the name “‘sum- mation tones.” The term “combinational tones” has been used to include both differential and summation tones. By means of tuned resonators Helmholtz was able to make both combi- national tones more clearly audible. He assured himself of their objective reality by observing the vibration of plates and resonators tuned to their frequencies. He distinguished between differential tones and beats. To quote Helmholtz, “when the places in which the two tones are struck are entirely separate and have no mechanical connection, as, for example, if they come from two singers, two separate wind instruments, or two violins, the reinforcement of the combinational tones by resonators is small and dubious. Here, then, there does not exist in the air any clearly sensible pendular vibration corresponding to the combinational tones, and we must conclude that such tones, which are often powerfully audible, are really produced in the ear itself.” In 1876 K. R. Koenig began the publication of a series of acoustical papers. In the German edition of one of Koenig’s papers the actual existence of differential tones was acknowledged, though “extraordinarily weaker than beat-notes,”’ but a few years later the French edition contained the following statement by Koenig: “No experiment has yet proved with certainty the existence of differential and summational tones’”(1). This started a controversy which continued for over half a century and engaged the attention of some of the greatest physicists of the time. Koenig stated in 1876: “Neither these combinational tones nor the beat-notes already described are reinforced by resonators.” Twenty-Seventh Annual Meeting 35 W. Preyer(2) used a set of tuning forks so delicately constructed that they had to be used during the quiet hours of the night. He found in every case in which a third fork responded to two forks sounded simultaneously that one of the combining forks alone caused the response. This indicated a re- sponse to a harmonic. In 1881 R. H. M. Bosanquet(3) described experiments with a resonator so arranged that external sounds were excluded from the ears. (Helmholtz had used resonators connected to one ear so that external sound was admitted in the other.) Bosanquet concluded: “In short, the difference-tone of Helmholtz, or first beat-note of Koenig, as ordinarily heard, is not objective in its character.” By 1895 the issue was almost decided against Helmholtz by the over- whelming evidence produced by Koenig, Bosanquet and Preyer. At this time Rucker and Edser(4) reported the results of an investigation in which a small mirror was attached to one prong of the responding tuning fork. The mirror was used in a Michelson’s interferometer arrangement so that a periodic vibration of the mirror amounting in amplitude to a half wave- length of light could be detected. They employed a double siren, which Helmholtz had found most effective in producing combinational tones, instead of two separate forks as the source of the combining tones. Although the results were somewhat dubious, they confirmed Helmholtz in maintain- ing the existence of combinational tones. Lord Rayleigh(5) stated in 1896 that he was at a loss to understand how two opinions are possible. He agreed with Helmholtz. In 1906 E. Waetzmann reported the detection of a difference-tone by | beats arising between it and another tone of slightly different pitch. He used a soap film over the opening of a Koenig resonator as the receiving apparatus. Later he used a loaded membrane(6) to simulate the action of the ear in its response to combinational tones. The Helmholtz-Koenig controversy subsided with no unanimity of findings. Major physicists disagreed upon the existence of combinational tones external to the ear. No one explained clearly the fundamental con- ditions under which they are produced, if indeed they can be produced objectively. (7) The Sideband Controversy. In 1928 the same issue arose in the field of radio communication. No one seemed to recognize that it was fundamentally the same question that physicists had debated in the realm of acoustics. For several years radio engineers argued about the physical existence of sidebands. These sidebands are frequencies caused by modulating a radio carrier wave. Frederick K. Vreeland(8) proposed to gain better fidelity in radio re- ception by the use of band pass circuits which had steep but flat-topped response curves. Vreeland pointed out that when both upper and lower sidebands are transmitted, each must be received in its entirety in order to avoid distortion. Highly selective circuits which discriminated against the sidebands appeared to cause distortion in radio telephony. 36 The Kentucky Academy of Science Promptly Professor G. W. O. Howe, editor of Experimental Wireless and Wireless Engineer suggested that Vreeland’s proposal be subjected to rigorous mathematical analysis. Professor Howe stated that “the transmitting station actually sends out waves of one definite frequency, but of varying amplitude and such a modulated wave has the same effect on a receiver as a wave of constant amplitude but of different frequencies, the different frequencies covering a range on either side (sidebands) of the actual frequency of the wave.’ A. W. Ladner quickly objected to the idea that modulation consists in changing the amplitude of a carrier frequency and cited experimental evi- dence of the tangibility of sidebands. Ladner emphasized that the spectrum of frequencies appears in the transmitter as soon as modulation occurs and that these frequencies are radiated to the receiver as objectively real waves. Some of Britain’s best radio experts participated in the discussion. Frank Aughtie cited the absence of a frequency of double that of the modulation frequency in the detected output of a receiver tuned to the second harmonic of a transmitter as evidence against the view of a modulated wave as a spectrum of frequencies when dealing with a valve. Ladner then asked how “‘suppressed carrier” or “single side band working” could be explained on the basis of a modulated wave as a single frequency. A. B. Howe answered Aughtie’s question by rigorous mathematics. Then came Robinson’s stenode radiostat receiver. Claims of remarkable selectivity without consequent distortion were made and these were challenged by Professor Howe. Sir J. Ambrose Fleming, noted inventor of the diode tube, expressed the view in an issue of Nature(9) that sidebands do not exist objectively. His view paralleled that of Koenig in the controversy over combinational tones. Professor Howe sharply criticized Fleming and his analysis of sideband reality. F. M. Colebrook(10) published experimental data on the response of a circuit to a modulated signal. Peaks of current in a selective tuned successively to carrier, lower sideband and upper sideband frequencies showed that a spectrum of frequencies existed. But Colebrook used a triode recitifier with his tuned circuit so that there was still the question of whether these sideband frequencies were produced by the receiving circuit or whether they existed in the modulated signal before it reached the receiver. The argument carried on in Experimental Wireless and Wireless Engineer became rather personal and at times bitter. Again there was no unanimity of conclusions. Studies by the Writer. The writer read the reports of these controversies with a great deal of interest. In 1927 he hoped to combine two frequencies in the light spec- trum in such a way as to obtain a frequency in the radio spectrum as the differential frequency. If light and radio waves were fundamentally the same kind of phenomenon, differing only in frequency, such an experiment cr oe Twenty-Seventh Annual Meeting Sy is theoretically possible. This assumes, of course, that both light and radio are wave phenomena. The maze of confused data and opinions on both the Helmholtz combi- national tones and the side bands led the writer to undertake first the settlement of these differences and to find exactly, if possible, the conditions giving rise to the derived frequencies in each case. During the years 1928 to 1933 this was accomplished. A paper(11) on experimental studies by the writer was published in 1935. The fundamental principle involved was found to be quite simple. The combination of two tones or frequencies, either acoustical or electrical, may or may not produce objectively real com- binational frequencies. If the parent frequencies are simply added, except through a non-linear medium, only beats are produced. Beat frequencies do not have objective reality. Koenig, Preyer and Bosanquet were merely adding frequencies. The result of adding frequencies is found mathe- matically as follows: Let A sin wt and A sin wet be two simple sinusoidal disturbances of the same amplitude which act simultaneously upon a linear aperiodic medium. ‘The resultant disturbance, y, is then the vector sum y—A sin wyt--A sin wet. From trigonometry y=2A sin [1/2 (witwe)t] cos [1/2(wi—we)t]. Also from trigonometric relations sin x cos y = 1/2 sin (x+y) + 1/2 sin (x—y). This would appear to give combinational frequencies, but if we let x= 1/2 (w1-++we)t, and y=1/2 (wi—we)t we get y=2A [1/2 sin 1/2 (witwotwi—we)t+1/2 sin 1/2 (wi+we—wi-+we) t] 5» or y=—A sin wyt+A sin wot. This is the same expression we started with. In other words, the equivalent frequencies turn out to be the original disturbances and nothing more, just as they do if developed in a Fourier series. If one frequency is caused to multiply another, as in their action upon a non-linear medium, the derived combinational frequencies are really pro- duced. To use the trigonometric relations again AiAzs 2 : 1A Ay sin wyt A» sin we t= i cos (wi—wWe) t— cos (wi-+wo)t. The frequencies wj1—we2 and wi+w» are the combinational frequencies. The lower one, wi1—we, corresponds to the differential tone of Helmholtz. It is a lower sideband frequency of a modulated radio wave while the higher one, wi-+we, is the summation tone of Helmholtz or an upper sideband frequency of a midulated radio wave. Modulation is simply the process which causes the multiplication of one periodic disturbance by another. In a circuit using Heising plate modulation, the product or multiplication of two frequencies, the radio carrier and the audio modulating frequency, is caused. In non-linear detecting circuits the multiplication of two fre- 38 The Kentucky Academy of Science quencies is caused by the action of the detector.(*) Other terms than simple products may be caused. For example, if the response of a medium, circuit, detector or apparatus of any kind is given by yoaf+bf?+cfH+ ....... : the simultaneous action of two disturbances, f, and fo will give TS) Ghia) bo he ide When the second term is developed we have b (f71-+2f1f2+ £2). This is seen to contain the squares of f; and fs as well as the product f, fo. When periodic functions are put in for f; and fs, the product f; fe gives the combinational frequencies and the second power terms give constants and harmonics. The writer has found experimentally that a resonant circuit does not respond when tuned to the difference in frequency between two separate oscillators. This is a case of simple addition of waves. It parallels the experiments of Koenig, Preyer and Bosanquet. A resonant circuit does respond when tuned to the sideband frequencies of a modulated radio wave. This experiment parallels the work of Helmholtz. Two loudspeakers could not be made to modulate each other either in air or by mechanical connec- tions. An electroacoustical wavemeter developed by the writer was used in analyzing the complex sound waves present. The most important contribution made by the writer was an experimental proof of sideband production with strictly linear circuit elements used throughout. With the aid of Dr. R. R. Ramsey, professor of physics in Indiana University, whose sustained interest and encouragement have been an inspiration to the writer, this crucial test was made. A spiderweb coil was vibrated in the radial field produced between two honeycomb coils. The axial component of the variable field set up by an A.C. generator in series with a D.C. generator through the two honeycomb coils was balanced out. The honeycomb coil was connected to a wavemeter which showed peak currents at the sideband frequencies. Is the Ear Linear? E. B. Newman, S. S. Stevens and H. Davis(12) reported in 1937 that the electrical impulses produced in the ears of cats and guinea pigs by sound stimulation are non-linearly related to the sound stimuli when the intensity reaches a sufficiently high level. By means of a suitable operation on the animal, access was gained to the round window and in some cases to the outside of the cochlea itself. Cochlear potentials were picked up by a wick electrode and fed into an amplifier whose output was studied by means of a wave analyzer. When two pure tones were impressed upon the ear, combinational frequencies were observed in the cochlear potential. Combi- national tones as high as the seventh order were detected in the response to pure tones of 700 and 1200 cycles per second. The first order sum and difference tones appeared at about 40 decibels above the threshold of audibility. The experimenters attributed the production of these com- binational tones to the non-linearity of the ear, more specifically to asymmetry of the middle ear. The non-linearity arises, they maintained, Twenty-Seventh Annual Meeting 39 when members of the ear are pushed beyond the limits within which Hooke’s law is obeyed. They regarded the asymmetry as largely due to tension of the muscles of the middle ear. Combinational Tones in Optics. In 1939 Sir William Bragg(13) cited the work of the writer, confirmed the findings, and proposed an extension of the application of modulation to explain the peculiar scattering of light known as the Raman effect. Bragg described an interesting method of producing modulation which shows clearly the difference between adding periodic disturbances and causing | one disturbance to multiply another. He focused light from a rectangular aperture upon the space between the prongs of two tuning forks placed at right angles to each other and in parallel planes. The area of the trans- mitted beam of light and hence its intensity was proportional to (a+A sin wit) (b-+B sin wot) where a and b are the sides of the rectangular image when the forks are motionless and A and B are the amplitudes of the simple harmonic motions of the respective forks. The beam of light fell upon a photoelectric cell whose output was amplified and sent to a loudspeaker. Analysis of sound _ from the loudspeaker showed the presence of combinational frequencies. But when the prongs of the forks were parallel, the intensity of the trans- mitted light was proportional to b(a-+-A sin wit-+B sin wet) and no combinational fre- quencies were observed. Concerning the Raman effect, Bragg offers the following explanation: “The incident light of definite frequency excites a forced vibration of the same frequency upon the molecule which it meets. If the magnitude of the response to the electric force depends upon some quantity in the molecule which is undergoing simple harmonic motion, as for example some relative oscillation of the atoms in the molecule, modulation occurs. Lines representing combinational tones are found on each side of the line repre- senting the original frequency in the spectrum of the scattered light.” In conclusion, we see that the production of combinational frequencies is an important general process in nature. It is as broad in its applications as the fundamentals of simple harmonic and wave motions. *There is no fundamental difference in the action of a “detector,” a “modulator” and a “demodulator” of radio waves. All accomplish the product of sinusoidal terms. (1) Helmholtz. Sensations of Tone. Ellis’ translation. Barton. Text Book of Sound. (2) Preyer, W. Akustische Untersuchungen. Jena. 1879. (3) Bosanquet, R. H. M. Proc. Phy. Soc. of London. May; 1881. (4) Ruker and Edser. Phil. Mag. 39; 1895. (5) Theory of Sound, vol II. 40 The Kentucky Academy of Science (6) Waetzmann, E. Annalen der Physik. July; 1913. (7) Ramsey, R. R. Science. June 7; 1935. Am. J. of Phys. Aug; 1940. (8) Vreeland, F. K. Proc. Inst. rad. Engrs. Mar; 1928. (9) Fleming, Sir J. A. Nature. Jan. 18; 1930. (10) Colebrook, F. M. Exp. Wireless and Wireless Engr. Jan; 1931. (11) ‘HazelS He Phil Mac) ian) 1935. (12) Newman, E. B., Stevens, S.S. and Davis, H. J. Acous. Soc. of Am. OcEe 19377 ; (13) Bragg, Sir W. Nat. 143; 542; 139. ON THE FOUR-DIMENSIONAL MECHANISM OF KNOWING LEsTER S. O7BANNON Agricultural Experiment Station University of Kentucky The universe is a four-dimensional flux of kinetic-atomic elements. The human being is a part of this flux and is continually changing the complexion of its atomic configuration to conform to the changing com- plexion of the whole. This acting on the part of the human being we call behavior. Knowing is a kind of behavior. Knowing is acting. Thinking is a kind of behavior; and so is working, sleeping and eating. Let us examine eating. First, we procure food from our environment. We take the food into our bodies. It is broken down into its elemental constituents and assimilated into the structure of the body. The atomic elements of the food com- mingle with the atomic elements of the body. ‘There is the building of body tissue to maintain the continuance of the body as a body. Why does the body maintain itself? Why does not the food rot; and the body disinte- grate? No one knows; but this much is certain: the assimilation of food is a directed act on the part of the body. It is a determined act. It involves a force, or impulsion, coming from behind to seize upon the food and mold it into body tissue. We say that energy is stored in the body. What does the body do with the energy? It uses the energy to obtain more food. The act of using the energy, or assimilated food, is telic. It is a purposive act; a means to reach an end. Question: At what point in this process did the disposition of the body change from deterministic to telic? Answer: The disposition did not change at all; it was both deterministic and telic at the same time. The body was not strung out on an imaginary line divisible into before and after; the body existed only as an event. This event was a change in the atomic configuration of the body caused by a potential difference between suc- cessive complexions. That is, we assume, or infer, that there was a potential difference; really, there was only the fact of change. ‘To imagine the change as having direction is to invent a geometry. We make use of i Twenty-Seventh Annual Meeting 41 this geometry when we divide a process into two phases and describe one as deterministic and the other as telic. As to the assimilation and utilization of food there is only the one event of metabolism. To call it anabolism and katabolism is merely to diagram it or analyze it according to a preconceived geometry, however obscure that geometry may be. Actually, there is only the continually enduring configuration constantly undergoing a change in complexion. If we think of the change as having been impelled, we apply one name; if induced, we apply another. The names themselves are inventions which derive their meaning from commonplace phenomena. We have no parts of speech, no symbolism, no geometry, with which we can describe an organic whole in all of its fullness. We can affirm the existence of an organic whole, and write a formula for it, but the moment we begin to describe it, we cut it to pieces. To cut it to pieces means to diagram it in terms of geometry. And so with the act of knowing. Knowing is a single four-dimensional event. In order to describe knowing there is no alternative to analyzing it into those elements or phases which parallel, coincide with, or intersect, the coordinates of the four-dimensional whole. In addition to the four-dimensional whole, there are at least five geometrical constructs which may be applied to knowing. These are: 1. The fourth dimension alone conceived as representative of a potential difference to be interpreted either as a driving force or as an attractive force. 2. Equilibrium reactions in the spatial field, so far as we can conceive these as being three-dimensional only. 3. Reactions in the spatial field combined with the fourth dimension conceived as representative of a driving force. 4. Reactions in the spatial field combined with the fourth dimension conceived as representative of an attractive force. §. A bias cut through the four-dimensional whole, unoriented with respect to simple space and time. The five familiar concepts which correspond respectively to the foregoing geometrical constructs are: 1. Affective tendencies, such as attention, will, growth potential, genetic energy, and so forth. 2. (a) Sensation; (b) Apperception. 3. Perception. 4. Conception. 5. The specious present. These latter concepts may be defined by relating them both ontologically to kinetic atomicity, and juxtapositionally with respect to their concurrence in the four-dimensional whole. 1. Affective tendencies are necessary logical inventions, or abstractions, which coincide ontologically with the fourth dimension. They are non- spatial, non-extensive, fourth-dimensional potentials which give direction to the changing complexions of spatial configurations. 42 The Kentucky Academy of Science 2. (a) Sensation is the awareness of contact with stimuli. It is three- dimensional only. Sensation involves only the impingement of stimuli, the spatial redistribution of the responsive elements within the organism, and the bare awareness of sense data, so far as awareness can be thought of as three-dimensional only. Sensation does not involve interpretation of the data either as to their source or their utility. 2. (b) Apperception is the fusion of sense data with the cognitive pattern of the individual. It is the cumulative phase of cognition, and involves only the spatial redistribution of those elements which account for the new disposition which results from the immediate sensory experience. Apperception is three-dimensional only and does not involve interpretation of the data. If we think of sensation as inward redistribution, then apperception is to be thought of as outward redistribution. 3. Perception is the act of evaluating and classifying sense data in accordance with effective tendencies dominant at the moment and inherited from the experience of the individual; that is, the ontogenetic inheritance. Perception involves the present and the past but not the future; that is, it involves three-dimensional space and a fourth-dimensional potential, the latter to be conceived as a positive driving force intrinsic in the genetic constitution of the organism. 4. Conception is the evaluating and classifying of apperceived sense data for future use; that is, as means to reach an end. It is a telic process as contrasted with the deterministic process of perception. It involves a fourth-dimensional potential conceived as a negative or attractive force drawing upon the apperceptive mass for the purpose of meeting a new situation. 5. The specious present is the duration of consciousness in the combined act of perception, apperception and conception. There is in reality only the single act of knowing. If we think of this act as involving the four- dimensional flux in all of its fullness, unanalyzed by means of the naive geometrical constructs of space and time, then the specious present is an increment of duration for the organism along a world line (cf. Jeans) of the four-dimensional whole. Within this increment of duration the organic configuration assumes a new complexion, the assuming of which involves the reception of peripheral stimuli and the distribution of the effects, these being spatial concepts, together with the organizing force of an intrinsic potential difference, this being a fourth-dimensional concept, so that within this increment of duration the organism possesses temporal thickness (the specious present) measurable along an imaginary line beginning with the first atomic element of the organic configuration and ending with the last, this last being also the first. Finally, knowledge may be defined as those accumulated increments of experience knitted into the pattern of the organism which alter, or are cap- able of altering, the conceptual classification of new perceptions. Twenty-Seventh Annual Meeting 43 ECOLOGY OF KENTUCKY FLOWERS: I. CYAYTONIA, MERTENSIA AND JEFFERSONIA Harvey B. LovELL Department of Biology, University of Louisville Few studies have been made of the anthesis and florecology of Kentucky wild flowers. In the present paper the changes through which the flowers pass during anthesis has been traced for three spring flowers, which are characterized by unusual and little known mechanisms. I. CLAYTONIA VIRGINICA L. (PORTULACACEAE) Claytonia virginica, the spring beauty, exhibits very marked proterandry, a fact first noted by Bessey(1) in 1873. Meehan (1876) believed that Claytonia(2) is regularly self-fertilizing due to the closing of the flowers at night bringing the stamens in contact with the stigma. Robertson(3) (1889) ridiculed this idea pointiing out that the stigma branches remain closed and incapable of fertilization, until after bees have removed the pollen from the anthers. Staminate stage. On the first day the flowers are staminate. The five stamens stand erect in a circle around the closed stigma (Fig.1,A). The activities of small bees remove the pollen rapidly in favorable weather. The chief visitor in this region is Andrena erigeniae, a small black bee 8 mm. long. The flowers close in the late afternoon. Pistillate stage. On the second day the stamens with their wrinkled empty anthers bend back against the petals. The three lobes of the stigma expand widely exposing the inner surface covered with numerous papillae (Fig. 1,C). When a pollen-covered bee alights on the flower she is almost certain to touch the stima bringing about cross-pollination. Sometimes flowers may open for a third day. The flowers contain nectar and nearly all the visitors could be observed probing for it between the ovary and ring of filaments or occasionally on the outside of the filaments. The nectar is secreted by the expanded bases of the filaments (Fig.1,B). It is easily accessible, the flowers being adapted to pollination by short-tongued bees. Delicate rose-colored lines on the petals act as nectar-guides. The anthers and pollen are also rose-colored during the staminate stage, and many insects go to them directly to collect the pollen. In the pistillate stage the color fades out. Robertson is certainly correct in concluding that Claytonia is regularly cross-pollinated. On rainy days, however, the flowers do not open and so the pollen is not removed. If it rains on the second day, and the stigmas open within the closed flowers, they cannot fail to come in contact with their own pollen. Self-pollination then may be expected to occur only during prolonged rainy weather. On April 29th two persons collected 69 insects from a patch of about 1000 flowers in one hour. The chief visitor was the oligotropic bee, 44 The Kentucky Academy of Science Andrena erigeniae which visits Claytonia exclusively, its period of flight and brood rearing corresponding to the blooming time of the plant. The complete list of insects collected in Jefferson Co., Kentucky, follows: they were all sucking nectar except some of the Andrenae. HYMENOPTERA. Long-tongued bees. — CERATINIDAE: Ceratina me- tallica Sm. 7 F, 1 M; NomapripaE: Nomada sayi Robt., 1 M; N. pygmaea Cr., 4 m; Short-tongued bees——Hatictiwar: Axugochlorella striata (Prov.),1 F; Augochlorella aurata (Sm.), 1 F; ANDRENIDAE: Andrena erigeniae Robt., 53 F suc. and collecting pollen. DIPTERA. SYRAPHIDAE: Syrphus Sp. 1 MERTENSIA VIRGINICA (BORAGINACEAE) Mertensia virginica (L.) Link, Virginia bluebells or cowslip, has flowers which show a tendency toward dimorphism, at least in regard to the length of the styles. The flowers are adapted to pollination by long-tongued insects and secrete abundant nectar from 4 nectaries at the base of the ovary. The plants grow abundantly in rich, shaded soil along streams. The bright blue flower clusters are very conspicuous, and the plant is so widely sought for its beauty that it faces extinction in this locality. The buds are pink, changing to blue as they open. The bell-shaped mouth of the flower is about 15 mm. across. It narrows abruptly into a tube 12 to 15 mm. long and only 2 to 3 mm. broad (inside), and the style occupies the center of this space thereby reducing it still farther (Fig. 2, A). Five stamens are inserted at the apex of the tube. Their anthers dehisce longitudinally, rolling backward to expose the yellow pollen clinging to their inner surfaces. The ovary consists of four distinct carpels alternating with four green nectaries (Fig. 2, C). The carpels are arranged in pairs on each side of the flattened base of the style, leaving more space for two of the nectaries, which are, therefore, correspondingly larger. The base of the corolla tube is lined with dense whitish hairs, which protect the nectar both from intruders and from evaporation. Nectar secretion is abundant under favor- able conditions, filling the tube for several mm. with a sweetish liquid which can easily be tasted. The flowers are protected from the rain by their slightly nodding position. They have a sweetish odor. Dimorphism. 1 can find only one brief mention of dimorphism in Mertensia. The local specimens, however, show a great deal of variation in the length of the style. Long-styled flowers have the styles extending beyond the mouth of the flower, exceeding the anthers by 4 or 5 mm. This is the more usual condition (Fig. 2, B). Short-styled flowers have the style 2 to 3 mm. shorter than the anthers (Fig. 2, A). There does not seem to be a corresponding change in the length of position of the anthers. Occasional specimens show intermediate positions for the stigma. The flowers are homogamous, the anthers and stigma maturing at about the same time, cross-pollination being rendered fairly certain by the dif- ‘ $ Fr Twenty-Seventh Annual Meeting 45 i A neclary-- i | nectary- A) Fig. 1, CLayTonta virginica: A. Staminate stage, x 2; B. stamen with nectary, x 4; C. pistillate stage, x 2. Fig. 2, MERTENSIA VIRGINICA: A. Short-styled flower, x 114; B.C. Long-styled flower, x 144. Ovary with four nectaries, x 6. Fig. 3. JEFFERSONIA DIPHYLLA: A. Pis- tillate stage, x 3; B. Homogamous stage, x 3; C. Autogamous stage, x13. 46 The Kentucky Academy of Science ference in length of these two organs. Anthesis lasts 2 to 3 days, but the flowers may persist for another day after the pollen has been shed. Robertson(4) (1891) has given a brief account of M. virginica but over- looked the tendency toward dimorphism. More recently (1928) he(5) has published a revised list of the visitors. Only very long-tongued insects can obtain the nectar. Robertson lists for Illinois 5 species of Bombidae, 4 of Megachilidae, 1 of Nomadidae, 3 of Euceridae, and 2 of Anthophoridae, all long-tongued bees. The honeybee visits the flower only for pollen, its 6 mm. tongue being too short to reach the nectar. He also reports 3 butterflies, 2 sphinx moths and two flies sucking nectar. Both at Carlin- ville, Illinois, and at Louisville, the ruby-throated hummingbird is a fre- quent visitor(6). Schneck (1891) observed Xylocopa virginica perforating the base of the corolla tube and honey bees sucking from the holes(7). JEFFERSONIA DIPHYLLA (L) Pers. (BERBERIDACEAE) Jeffersonia diphylla, the twin-leaf, has developed a most unusual mech- anism for self-pollination and in this locality at least is rarely cross- pollinated. The solitary scapes bloom in the deep woods in late March and early April and then their delicate white petals fall. Insects are rare and erratic so early in the year, especially if the weather is cold and wet. The large white flowers have 8 petals, 15 to 18 mm. long, which render the flowers very conspicous against the dark background of the forest floor. The pistil consists of a green ovary, soon becoming gibbous and an ir- regularly-lobed sessil stigma. A circle of 8 stamens are inserted opposite the petals, the anther being 7 mm. long to 3 mm. for the filament. The pistil and stamens are about the same length. The flowers are briefly pro- terogynous soon becoming homogamous. 1. Pistillate stage. "The stigma appears recptive in flowers while the extrorse anthers are still closed (Fig.3,A). The petals stand nearly upright forming a tube. Any insect alighting in the flower at this stage would be certain to strike the large stigma. 2. Homogamous stage. In a day or two the covers or valves of each anther-cell break loose from the bottom and gradually roll up. The flower is now homogamous (Fig.3,B). The whole flower continues to increase in size during these early stages, the petals lengthening 3 to 4 mm. and the pistil 1 to 2mm. The anther-covers roll up very slowly, some of them lagging behind, constantly exposing a fresh supply of pollen. 3. Autogamous stage. Finally after several days the stamens bend in toward the pistil and the anther-covers turn over the top of the anthers, partially unrolling as they do so, until the covers comé in contact with the stigma (Fig. 3,C). Since pollen was still clinging to the anther-covers in the specimens which I examined, this mobility seems certain to bring about self-pollination. The petals expand more widely during this stage. Graenicher (1906), who made some observations upon Jeffersonia in Wisconsin, thought that self-pollination was brought about by the pressure of the petals on the stamens as they close at night. I think he must have Twenty-Seventh Annual Meeting 47 been mistaken, as the bending in of the stamens and looping over of the anther-covers give every indication of being spontaneous movements. In Louisville I collected only a single beetle and on several occasions observed large patches of Jeffersonia without finding a single insect visitor. In Milwaukee, Wis., Graenicher(8) has reported 9 species of bees collecting pollen, mostly small Halictine bees which he believed occasionally effected cross-pollination. Why is so conspicuous a flower rarely visited by insects? This is due partly to the lack of nectar and partly to the early blooming time. Any visits would be made either accidentally by insects in search of nectar (and would probably not be repeated) or by insects collecting pollen. The lack of nectar and the self-pollination mechanism may be regarded as a degenerate specialization brought about by mutations. The ability to perform self-pollination made the presence of nectar no longer of survival value. As is now well known, regressive mutations which are common for all organs, soon accumulate in large numbers for organs or structures which no longer are necessary for the preservation of the species, and often lead to the disappearance of such structures. The autogamous habit also makes possible the early blooming of Jeffersonia when the weather makes insect visits unreliable. Although autogamy may be an efficient method of setting seed, it greatly decreases variability. There is only one species of Jeffersonia in North America and only two in the world. Should environmental conditions change, the twin-leaf would probably be unable to adapt itself and would become extinct. (1) Bessey, C. E. 1873. Sensitive stamens in Portulaca. . .Am. Nat., vu, 464-465 (2) Meehan, Thomas. 1876. The ‘Sleep of plants’ as an agent in self- fertilization. Proc. Acad. Nat. Sci., Phila., p. 84. (3) Robertson, C. 1889. Flowers and Insects. IJ. Bot. Gaz., xiv, 177. (4) Robertson, Charles. 1891. Flowers and insects. Trans. St. Louis Acad. of Sci., v, 580. (5) Robertson, Charles. 1928. Flowers and insects. The Sci. Press, Lancaster, Penn. (6) Schneck, J. 1887. How humblebees extract nectar from Mertensia virginica. Bot. Gaz., xu, 111. (7) Schneck, J. 1891. Further notes on mutilation of flowers by insects. Bot. Gaz., xvi, 312-313. (8) Graenicher, S. 1906. Some notes on the pollination of flowers. Wis. Nat. Hist. Soc., rv 12-21. 4G The Kentucky Academy of Science A STUDY OF THE NESTING BIRDS OF A SEVENTY“AGRE AREA IN ROWAN COUNTY, KENTUCKY - (abstract) R. W. BarBour Morehead State Teachers College During the spring and summer of 1938, the writer, assisted by Thomas Marsh and John Rice undertook a survey of the nesting birds of a seventy — acre area near Morehead, Rowan County, Kentucky. The area was chosen — with a view to including a wide variety of habitats. Habitats ranging — from uncut meadow through a dense swamp-like area to a hill covered with — second growth timber were included. E A total of forty nests, representing fourteen species, were observed. The — forty nests contained a total of 123 eggs, of which 55, or 44.72 per cent — hatched. This loss was due to predation, desertion, storms, infertile eggs, — and other factors. q Of the 55 young that were hatched, 43, or 78.18 per cent left the nest under their own power. ‘Thus, 34.96 per cent of the total number of eggs — laid produced young that left the nest. > Fourteen, or 35.00 per cent of the forty nests produced young that left — the nest. Of the twenty-six nests that were destroyed, twenty were de- stroyed before hatching, and the other six were destroyed at varying lengths — of time after the eggs had hatched. ; Nests of the following species were observed: Acadian Flycatcher, Tufted — Titmouse, Catbird, Robin, Wood Thrush, White-eyed Vireo, Red-eyed — Vireo, Blue-winged Warbler, Oven-bird, Kentucky Warbler, Yellow- — breasted Chat, Hooded Warbler, Eastern Cardinal, and Red-eyed Towhee. TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE (Quarterly Series) AFFILIATED WITH THE A. A. A. S. VOLUME 9—NUMBER 3 DECEMBER 1941 TWENTY-SEVENTH AND TWENTY-EIGHTH ANNUAL MEETINGS 1940-1941 ee CONTENTS Page Mercademy Officers, for 1940-1941 0 su0. i e 49 Official Reports Aléredy Brauer.) Secretary. fs A ia5 fe a Saas Ree Te Oem 50 Wis]: Moores (Treasurer i) 6k kk ea a he ee eR la 53 Enzymic Purification of Antitoxin Edward Kass M. Scherago Re! deta), Wheciwietumeny retain 2S) ene Neuve 4 aR te UN 55 Reaction of n-Butylmagnesium Bromide with N-Ethylisatin Watch Ge SUMIplen ns! se piri: dae datteded Mo ee nate pee ES 61 The Role of “c” in Systems of Units TR dhe ME NN RS saypeme ESP Wea Wrarburtom) 2 hii) hyiihiy Pees LEON INS io i} [764 {Inversion Applied to Conics ‘S ; ON Sister Charles Mary Morrison ... \ Roa APR 7-5-4049 68) Genetics of Intelligence \ ps Ws Edward Newbury. !)\... 0445.00 AT owes MUS ew. 3 STS (GSES Page ein a ear is XR ne RRR Ban SST. «79-80 eee es Application for entry as second-class matter is pending TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE EDITORIAL STAFF Charles Mlirey ia) e Murray, Kentucky .... Editor-in-Chief Charles Barkenbus ....... Lexineton, Kentucky - 2.9.) 3. Chemistry pies MB Fake iron eae eke ie ae Morehead: Kentucky 4. 2° =e Physics EWE Cookie as isin Danville, Kentucky .......... Bacteriology NW ROY Tatutehersom) 0.0.2). Befea, (Kentucky 0) .ae an Mathematics Jobin Kuiper) 5...) er Lexington, Ky. .... Psychology & Philosophy ty Be ovell ie oy. aes. Louisvalle, Wentucky =.) 1 see Biology AM @uMicBarlan ny ol aioe. 2: Kexington, Mentucky) 0/03) ssa Geology Manuscripts. The Transactions must be limited to the proceedings of the annual meetings of the Kentucky Academy of Science and to original manuscripts pertaining to science. Under present financial limitations each article must be limited to approximately five pages of the Transactions. Manuscripts are subject to the approval of the Editorial Staff. Manuscript material may be submitted to the Associate Editor of the subject covered or to the Editor in Chief. Exrra-Cost Features. ‘The extra cost of special features such as cuts, graphs, tables, etc. above the text-run price of $2.65 per page must be borne by the contributor. The Editorial Staff will advise contributors concerning the extra cost of special features upon receipt of manuscript. [Illustrations to be included in an article should accompany the manuscript if possible, or, if sent in separate package should be properly labeled as to the article in which they are to occur. Proor. Galley proof will be sent for approval of contributors. The proof should be returned promptly to the Berea College Press, Berea, Kentucky. REPRINTS. Reprints are furnished at publisher’s prices by negotiating directly with Berea College Press. Price quotations on reprints are sub- mitted with the proof. Orders for reprints. should accompany the proof to the Berea College Press, Berea, Kentucky. SUBSCRIPTION Rates. The Transactions is sent without additional ex- pense to all members of the Kentucky Academy of Science who are not in arrears for annual dues. The annual subscription rate for non-members is $2.00 in the United States and Canada, $2.50 in foreign countries; single number 75 cents. One volume of four numbers appears each Academy fiscal year. BusINEss CORRESPONDENCE. Remittances and correspondence concern- ing subscriptions, extra costs, and other financial matters except reprints should be addressed to Chas. Hire, Murray State Teachers College, Murray, Kentucky. BEREA COLLEGE PRESS, BEREA, KY. all By 3S B37 Twenty-Seventh and Twenty-Eighth Annual Meetings 49 j OFFICERS FOR 1940-1941* (PEST CIESIE tz gage aed 9s SA ee ast Aa on er Oe Chas. Hire | LE SEN SSTVG IS 8 al A nei eae i ia ice Cee ae a ae G. B. Pennebaker ELSES <5) 32 CONS ae SI ei ce ge een ger aS Alfred Brauer || PESUITSI a ROR RN CS alec EIR RO an Wm. J. Moore Representative on A.A.A.S. Council .............. Austin R. Middleton Councilor to Kentucky junior) Acadenty ete Anna A. Schnieb Peeroutearoresidentss hye eit ue eee A. W. Homberger DIVISIONAL OFFICERS FOR 1940-1941 “LIDIDS Sy a ee ee eo Gham nie? yang Shr one W. D. Valleau Morris Scherago SECEEEALIES wre teta eee ies disieal a Gy L. A. Brown Wm. H. Stark eiemistry .............. @ivaigmiany eae! sg ce ce ose W. H. Keller SEChebAGy a Yi sve eerie as: W. S. Hodgkiss Beolocy, and Geography ...Chairman .:.:.2:.....:... R. E. Stouder Secretar yan tc tt ome nes teceees V. E. Nelson Mitthematics ............. Gharrmanee i ae H. A. Wright SECLCCAGY eta! See hry he Ne D. E. South BEASIESA Sy cycle Chainmanies< 0! Gye wr oe whee R. A. Loring SCCLECAD VEN tee crs ee tks a ha ... Jarvis Todd Philosophy and Psychology..Chairman .................. Paul L. Hill SECKetan ye te ue rete is G. B. Dimmick *Officers for 1939-1940 are listed in Number 1 of this Volume. Ie eo en 50 The Kentucky Academy of Science TWENTY-EIGHTH ANNUAL MEETING OF THE KENTUCKY ACADEMY OF SCIENCE HELD AT RICHMOND, KENTUCKY APRIL 25TH AND 26TH, 1941 The first business session was called to order by the President, Charles Hire, at 1:30 P.M. The regular order of business was followed namely, 1. Presidents intro- ductory remarks, 2. Secretary’s report, 3. Treasurer’s report, 4. Report of auditing committee, 5. Report of the Councilor of the Junior Kentucky Academy of Science. SUMMARY OF REPORTS Secretary’s Report 1. Announcement of the decision of the Executive Committee that the Transactions would henceforth be published in quarterly form, begin- ning with Volume 9, No. 1, and containing the transactions of the Twenty- Seventh Annual Meeting. 2. Reorganization of the Division of Geology and Geography into the Geological Society of Kentucky, a branch of the Geological Society of America. Under the provisions of the reorganization plan, Members of the society automatically become members of the Kentucky Academy of Science, the treasurer of the society paying the annual dues of the members to the treasurer of the Academy. ‘The plan carries the approval of the Executive Committee of the Academy. 3. Organization of: the bacteriologists of the State into the Kentucky Branch of the American Society of Bacteriologists. This Society seeks af- filiation with the Kentucky Academy of Science. 4. A recommendation for the reorganization of the Board of Directors of the Academy, with an election of a new slate of directors. 5. A recommendation that the Membership Committee for 1941-1942 be instructed to attempt to enroll a greater number of the State’s A.A.A.S. members in the Academy membership, and conversely to encourage Academy members to become affiliated with the A.A.A.S. whenever possible. 6. A report on the Academy membership is herein given as follows: Total membership, 346; National members, regular,, 78; Corresponding members, 8; Honorary members, 13; Emeritus members, 2; Life members, 3. Signed, Alfred Brauer, Secretary. Twenty-Seventh and Twenty-Eighth Annual Meetings 51 Report of the Councilor to the Junior Academy (Dr. Anna A. Schnieb) Eighth Annual meeting of the Kentucky Junior Academy of Science, held at Danville High School, Danville, Ky., April 11 and 12. Attendance was approximately 450. Fifteen clubs brought sixty-five exhibits. During the year 9 new clubs were organized. Of these, 5 were class “A” clubs and 4 were class “B.” One club was lost, due to replacement of science teacher by an athletic coach. Total number of clubs, 41. Total membership, 938, including 45 sponsors. Largest Clubs: Bellevue Sr., 100; Maysville, 34; Paint Lick, 34; Bellevue Junior, 33. Clubs of High Distinction: Bellevue, clubs of 100 and 33 respectively; Covington, quality of articles presented in Bulletin; Danville, solicited $100.00 and planned annual meeting; Kirksville, quality of research; Lan- caster, number of articles presented in Bulletin; Maysville, all-around efficiency, excellent cooperation; Morehead, organizing new clubs. Some outstanding accomplishments: Nine new clubs organized; increased number of laboratory research problems; five six-page issues of Junior Science Bulletin published; increased number of clubs making special fi- nancial contributions; excellent attention and outstanding discussions at the Annual Meeting. Officers of Junior Academy for 1941-42: President, Thomas Oelrich, Danville; Vice-President, Beverly Miracle, Barbourville; Secretary, Herman Neeley, Somerset; Treasurer, Thomas Cole Phelps, Central High, Richmond. Report of Morris Scherago (Delegate to the S.A.A.S., Mobile, Ala., Mar. 20-22) Dr. Scherago reported on the organization meeting of the Southern Asso- ciation for the Advancement of Science. The report is filled in the archives of the Academy. The Kentucky Academy of Science as a scientific organization in this southern geographical region, voted to cooperate with the S.AA.S. in its efforts to promote scientific research in the South. The Second Business Session, was called to order by President Charles Hire, 8:00 A.M. April 26th. The report of the representative to the A.A.A.S. was heard at this time. It was approved and ordered filed. Dr. G. B. Pennebaker, Chairman of the membership committee presented the following list of nominees, and these were subsequently elected: New Members Charles F. Bortner, Expr. Station, Lexington J. H. Bradshaw, Russell, Ky. Charles Crum, Covington 52 The Kentucky Academy of Science Dr. Robert L. Driver, Univ. of Ky., Lexington Ralph Gelder, American Rolling Mills, Ashland Dale S. Gerster, Transylvania College, Lexington Dr. Harold T. Glover, Eastern State T.C., Richmond Creed Grumbles, Ashland H.S., Ashland Alton M. Harville Jr. Graduate Student, Univ. Ky., Lexington Dr. H. T. Hazel, Ashland Junior College, Ashland Lawrence Henson, Expr. Station, Lexington H. H. Lafuse, Eastern $.T.C., Richmond George C. Patterson, Murray State T.C., Murray Stacy B. Randle, Expr. Station, Lexington James L. Rose, Expr. Station, Lexington Roy G. Smith, Ashland H.S., Ashland Dr. James A. Stoops, Ashland Junior College, Ashland W.C. Templeton, Jr., Expr. Station, Lexington Jack R. Todd, Univ. of Ky., Lexington Dr. Orville Wheeler, Supt. of Schools, Ashland Dr. William S. Wilson, Georgetown College, Georgetown. Research Grants of the A.A.A.S. President Hire announced the placement of the A.A.A.S. research grants as follows: Mrs. Katherine Carr, Morehead S.T.C., $25.00 Dr. B. B. McInteer, Univ. of Ky., $25.00 Dr. Ward C. Sumpter, Western Ky. S.T.C., $25.00 (In each of the above cases, this was the amount requested) Officers of the Academy for 1941-1942 Dr. R. H. Weaver, reporting for the Committee on Nominations, pre- sented the following slate of officers which was Su bsegaey elected: President, Dr. G. B. Pennebaker, Morehead Vice-Pres., Dr. J. T. Skinner, Western S.T.C., Bowling Green. Secretary, Dr. Alfred Brauer, Univ. of Ky., Lemneera (reelected ) Treasurer, Dr. Wm. J. Moore, Eastern S.T.C., Richmond (reelected) Rep. on Council A.A.A.S., Dr. Austin R. Middleton, Louisville (reelected ) Resolutions Dr. A. W. Homberger, Chairman of the Committee on Resolutions of- fered the following, which were adopted: 1. That the members of the Kentucky Academy of Science take this opportunity to thank Dr. Schnieb for her untiring efforts in her work with the Junior Academy, and we pledge our continued support; pereoy y7L ore 0100°0 ¥80°0 On €{¥c'0 VSS £8 ED SE ie Zay SInoy 96 C9 y9t 96000°0 Z¢10 (Gh Jae ¥£8°0 Ore trl CS cl? sInoy 96 C'S €r1 0100°0 try 1'0 Ss og 00°T Woe evI CLs ere, simoy 7Z 64 6ST ~100°0 «8610 Vis vel Sar L£91 et ae Iv°C STROM Sir 8°¢ sit 71000 1020 Jig 091 o> £91 al PS°7 sinoy $Z OT O'rT Z100°0 80¢°0 0 Ge 0 C81 0 0$°7 9 eIpouy] Ge tS) cay Sy ~ (2) ie) rg er Be dab e eee FE BOs male. ct. BOO fe gt Bach At Sr SQ Glen igs S Sow gees a eee ae os ae te aoe ie Be ge ae ee ct to ete 0 A c ‘al ‘ai sees eo ee ie eee 2 3 ‘ Z Zz aqvordng ul saskjeuy ‘DoLt—o0'y Hd vusejd urxoyjue eisayiydip uodn aseqseip avzts0 “p 9 5°0 FO 399FFq Twenty-Seventh and Twenty-Eighth Annual Meetings 61 Van der Scheer, J. and Wyckoff, R. W. G. Proc. Soc. Exp. Biol. & Med. 43; 427; 1940. Van der Scheer, J. Wyckoff, R. W. G. and Clarke, F. H. Jour. Immunol. 39; 65; 1940. Foster, J. C., Scherago, M. and Weaver, R. H. Jour. Bact. 39; 35; 1940. (8) Braun, H. Zeits. fur Immun. und Exp. Ther. Originale. 3; 531; 1909. Marrack, J. and Smith, F. C. Brit. J. Exp. Pathol. 12; 30; 1931. Marrack, J. and Smith, F. C. Ibid. 12; 182; 1931. Smith, F. C. and Marrack, J. Ibid. 11; 494; 1930. (10) Heidelberger, M. and Kendall, F. E. J. Exptl. Med. 61; 559; 1935. (11) Marrack, J. R. and Smith, F. C. Proc. Roy. Soc. London B. 106; 1; 1930. (12) Humphries, J.C. Scherago, M. and Weaver, R. H. Jour. Bact. 41; 64; 1941. m3) Pick, E. P. Beitr. Chem. Physiol. Path. 1; 351; 1902. mi) Pope, C. G. Brit. J. Exp. Pathol. 19; 245; 1938. Rope €. G.. ibid, 205 132: 1939: Pope, C. G. Ibid. 20; 201; 1939. (15) Parfentjev, I. A. U.S. Patent 2, 065, 196; 1936. (16) Coghill, R. D., Fell, N., Creighton, M. and Brown, G. Jour. Immunol. 39; 207; 1940. (17) Zinsser, H., Endors, J. F. and Fothergill, L. D. Immunity Principles and Application in Medicine and Public Health, Chapter XIV. MacMillan. New York. 1939. (18) Hutchison, A. Brit. Med. Jour. 1939; 384; 1939. (19) Sandor, G. Comptes Rendus Soc. Biol. 130; 840; 1939. Sandor, G. Ibid. 130; 1187; 1939. (20) Weil, A. J., Parfentjev, I. A. and Bowman, K. L. Jour. Immunol. 35; 399; 1938. (21) Goldie, H. and Sandor, G. Comptes Rendus Soc. Biol. 130; 1530; 1939. (7 —S (9 — THE REACTION OF n-BUTYLMAGNESIUM BROMIDE WITH N-ETHYLISATIN Warp C. SUMPTER Department of Chemistry Western Kentucky State Teachers College, Bowling Green, Kentucky Hill and Sumpter (1) obtained 1-ethyl-3-n-butyl-3-hydroxyoxindole (1) thru the interaction of N-ethylisatin and n-butylmagnesium bromide. In view of the work of Kohn (2) and of Myers and Lindwall (3) on the re- 62 The Kentucky Academy of Science actions of N-alkyl isatins with phenylmagnesium bromide the reaction is being reinvestigated. I. When n-butylmagnesium bromide was added to a solution of N- ethyl- isatin in benzene and excess of the Grignard reagent avoided compound I was obtained in good yields. Identity of the product with the compound prepared by Hill and Sumpter was established by analysis and by melting point methods. When N-ethylisatin was added to an excess (four molecular proportions) of the Grignard reagent, compound II, 2,2-di-n-butyl-1-ethylindole-oxide- 2,3, was obtained. Varying quantities of compound I were also obtained in this reaction when the reaction period was of short duration. With reaction periods of twenty four hours or longer compound I was not obtained. Myers and Lindwall did not obtain the phenyl analog of I when N- ethylisatin was added to excess of phenylmagnesium bromide even when the reaction periods were of short duration. The slower reaction of n-butyl- magnesium bromide as compared with phenylmagnesium bromide is in keeping with the relative reactivities of the two Grignard reagents as reported by Gilman (4). In the preparation of compound JI the mag- nestum complex (from N-ethylisatin and n-butylmagnesium bromide) was hydrolysed in the customary manner with a mixture of ice and sulfuric acid. The ether layer was separated, dried over sodium sulfate, the ether removed and the product distilled under reduced pressure. The crude product, a red liquid, was collected between 170-180° at 5 mm. On stand- ing crystalline material separated and was collected. The liquid was again distilled, practically all of the product coming over between 173-176° at 5 mm. Analyses for nitrogen agree with the value calculated for II. a Sie cae C-OH | Se Bee cats F2Ms qa. a —— CHOH eee | Ic. GH, 0 co yy \n7 CH, o.45 Twenty-Seventh and Twenty-Eighth Annual Meetings 63 The solid fraction consisted largely of N-ethyldioxindole (III). The formation of III in this reaction is not surprising in view of the well known reducing action of Grignard reagents (5). The N-ethyldioxindole ob- tained in this experiment was identified by comparison with a known sample by melting point methods. A small quantity of a second crystal- line substance melting at about 175° (quantity too small for repeated crystallization) was also isolated from the solid fraction. This latter sub- stance may well be 3,3-di-n-butyl-1-ethyloxindole (IV). Myers and Lind- wall obtained small yields of 3,3-diphenyl-1-ethyloxindole (VI) in their preparation of 2,3-diphenyl-1-ethylindole-oxide-2,3 (V) as outlined above. Compound VI was formed thru rearrangement of V by the action of the hydrolytic agent (sulfuric acid). Cw i\c H [9 ei, oe ~ AN We Bie “gs C.H C.,H.. cma V. 2 9 o Vio Solutions of compound II in the common organic solvents are light yellow in color and exhibit the same green fluorescence shown by solutions of compound V of Myers and Lindwall. Compound II is a liquid while V is a crystalline solid. Freshly distilled samples of compound II are red in color. On standing this color gradually changes to a golden yellow. This yellow liquid distills at the same temperature (173-176° at 5 mm.) and the distillate is again red in color but on standing gradually changes again to the golden yellow. The investigation is being continued with the view of isolating the second crystalline substance in sufficient quantity for analysis and characterization. It is hoped that for purposes of comparison compound IV can be syn- thesized thru the application of methods developed by Julian (6) for the synthesis of 1,3,3,trialkyl oxindoles. Attempts to bring about the rearrange- ment of II to its isomer IV thru the agency of those reagents usually ef- fective in bring about rearrangements of this type (sulfuric acid, acetic acid and iodine, acetyl chloride, etc.) are also contemplated. Complete experimental details will be presented later when further pro- gress has been made. (1) Sumpter, Ph.D. Dissertation, Yale University 1930. (2) Kohn and Ostersetzer, Monatsh. 34; 789; 1913. (3) Myers and Lindwall, J.A.C.S. 60; 2153; 1938. Myers, Ph.D. Dissertation, New York Univ. 1937. (4) Gilman, St. John, St. John and Lichtenwalter, Rec. trav. chim. 55; W754) 805 1936. 64 The Kentucky Academy of Science (5) Whitmore et al., J.A.C.S. 63; 634; 1941. (6) Julian, Pikl and Boggess, J.A.C.S. 56; 1797; 1934. THE ROLE OF “c” IN SYSTEMS OF UNITS F. W. WarBUuRTON > Department of Physics University of Kentucky, Lexington, Kentucky Many people regard the choice of units as a mere matter of convenience, and remain indifferent to the results of poor choices. They forget the power of habit and fail to recognize the psychological effect of repeating as true, day in and day out, statements and inferences which are not quite ac- curate or logical. Indeed when one studies the various points of view of men working on units, one cannot fail to recognize that a man’s ideas on units depend on how he habitually uses them or on how he was taught. It takes a deliberate effort to cut loose from the restrictions of preconceived ideas. In fact, one of the editorial critics of my previous paper“) claimed to see nothing sacrosanct about including the speed of light c=3x101° cm/sec in the units. Of course after that I have to emphasize all the more the need of including c. In the long list of half true or half complete statements which do as much harm as good, we may place another common phrase. In the same category with (a) “force between two charges” which confuses the Freshman when one means not any force at a point lying somewhere between the charges but the force on one charge in the presence of the other; (b) defining H as “force on a unit pole” when its predominate uses are the crosswise force on a current in a motor, the deflection of cathode rays, induced e.m.f. and radio waves, none of which apply directly the “force on a pole”; (c) “lines of force” when the force on a current the students use in the galvanometer and motor are not directed along these lines; (d) “cutting lines of force” when one means the real effect of a moving conductor or a changing pri- mary current, rather than stretching an elastic imagination to cut fictitious rubber bands; (e) “electromotive force” itself which is not force but work per charge; (f) ““magnetomotive force” which was not force but work per pole; and (g) “all circuits are closed by Maxwell’s displacement current,” which is not literally true when current is defined as passage of electrons per unit time and there is no such charge moving across the space between condenser plates in vacuum; we may place an eighth one (h) “the speed c is the ratio between the units” since rather it is a factor giving the relation- ship between electric and magnetic forces in any system of units. Bridgman in his Dimensional Analysis points out that the introduction of c simplifies one case and later remarks that consideration of dimensions has not advanced the understanding of electric relations very much. Bridgman does not go far enough. Showing early in the game that dimensionally a a a Twenty-Seventh and Twenty-Eighth Annual Meetings 65 factor c must appear in all systems of units, materially clarifies the situation. We know that f, = a qiqe/r? expresses the electrostatic force in any system of units, the constant a having a different value for different systems. Also the force on a moving charge is given by fm = b qoavesinaSqivisin6/r? in any system, using different values for b with different systems, so the total force is f =a qoqi/r? + b gevesinadqivisind/r?. Defining a new constant c by c= \/a/b, so b = a/c?, this becomes f = a[qoqi/r? + qevesina (1/c?2) Sqivisind/r?] where c must be a speed in order for both terms of the equation to have the same dimensions. Also experimentally c=2.998 x 108 meter/sec, the value of the speed of electromagnetic effect. Just coincidence? Oh no, of course not! The electrostatic system is chosen by omitting a, formally setting a=l and dimensionless, Then fi — qoqi/t? 4+ iglosina ( 1/c?) fh i,d]ysinO/r? In the electromagnetic system b is omitted. Then f — c2qoq1/r? + iglosina f iydl,sin@ /r? In Gaussian units a is omitted and one factor c is included under each i, so that i/c in Gaussian units is equal to i in electromagnetic units, thus preserving the electromagnetic units of B and H, the gauss and the oersted. The force is © f = qoqi/r? + (islo/c) sina f izdlysinO/cr?. The Heaviside-Lorentz units differ from the Gaussian units by a factor of \/4r. In the meter-kilogram-second system of units, the ampere and the volt are retained, (icmu=iamp/10) and the force in newtons is found from ~ faewtons —= Wioutes/!meters, 80 Substituting in the equation for force in electro- magnetic units fem tons = 02 basnes = c2Q.Q,/107r2+ Islosina fIidlisin6/107r?, where c is now 3 x 108 meter/sec. This explicit form, I call the normal form of MKS units. In the socalled rationalized form of MKS units, when qj is restricted to include only charges on conductors excluding the polarization charges q’ in the medium, and with q/x written for q + (—q’), the various factors c?, 107 ergs per joule and x are further lumped together in e=107%«/4zc?. In place of the total current I,, the sum the conduction current i and the am- perian current i’ (around the cylindrical surface of a cylindrical magnetized bar which takes the place of the fictitious poles on the ends), one writes ji, and lumps the various factors together in 747/10". Many people use p» for 66 The Kentucky Academy of Science 42/10". The new symbol y, which partially resembles » upside down is sug- gested to avoid confusion. (The joke about this is, first, that » can well go in the numerator even for the force of poles as Sommerfeld suggests, and per- haps our usual law f—=mm//pr? has had p in the wrong place all these years; and second, that in avoiding Professor Michenor’s symbol v for this quantity because it might be confused with the late Professor Kennelly’s use of v for a different quantity, I used y because it was convenient on the typewriter I was using, and I had no idea it would be referred to as that symbol in the A.A.P.T. committee report.) The resulting equation in “rationalized” MKS units is f = :qoqi/4rer? + ivglosinafyisdlisino/42r2_ newtons. In the atomic system of units, the charge on the electron is chosen as unity, so charges q are replaced by the numbers z of electronic charges, and f = 2921/12 + (Zevesina/c) Sz1visin6/cr? dynes. This becomes a two dimensional system since force becomes expressed with the dimensions of r—?. Now let us go back to our definition of current. From the time our great great grandfathers were little tots, current has been defined as i=q/t even when f=qoqi/r?. Also we have had for a long time the electromagnetic unit of current defined as the current producing unit electromagnetic field per radian of arc at the center of a circular coil. This field is readily expressed as equal to the transverse force on unit length of unit current is at the center of the circular coil and due to the current i, in the coil. This definition has the distinct advantage of its close relation to the experimental determination of the ampere in the current balance. We can combine these two ideas nicely to obtain a single complete theoretical system of units, merely by defining current as i—q/ct. Then q is expressed in statcoulombs and is that charge which repels a like charge according to qoqi/r?, and i is expressed in abamperes and is that moving charge which exerts a magnetic force on another moving charge according to the relation fn = ielesina fisdlysin6/r?. The total electrostatic and magnetic force is written as £ = qoqi/r? + iolysina fisdlysin6 /r? = qoqi/r? + (qoeve/c) sinaSqivisin6/cr?, the factor c appearing in the ratio v/c when magnetic forces are expressed in terms of moving charges but not appearing in the magnetic equation in currents. The physical concept that magnetism is the effect of moving charges only, is fundamental, and it seems appropriate to call this system of units the basic system. When we turn to the question of why c appears, we run straight into the philosophy of physics and into interpretations of relativity. I do not —_ Twenty-Seventh and Twenty-Eighth Annual Meetings 67 pretend to be any authority on these matters, but I am pretty sure that in the future we will come to look upon magnetic forces as a form of rel- ativistic correction to electrostatic forces. Let us review a bit of history. Mechanics, heat, electricity, magnetism and light were once pretty well separated. They are well united now and the trend toward unity is still going on Statistical mechanics takes care of heat, as entropy being a statistical probability may without difficulty be taken to be a pure number, and then temperature has the dimensions of energy, and the three mechanical units are sufficient for heat. QOecrsted’s discovery connected magnetism with electricity. Ampere, Weber, Riemann and others made considerable ad- vances with relative motion and reciprocal action of one charge on another, obtaining waves along wires and a propagated potential. But it was Maxwell, a man of our own tongue, who saw that the work to carry a crude pole around a current could be retained provided he assume a dis- placement current between condenser plates. However there were many difficulties cropping up continually. Maxwell’s theory violated reciprocal action of moving charges, and claimed absolute motion in an elastic ether. Lorentz neatly dispensed with the need of a medium other than propagation through vacuum, by explaining the dielectric constant and permeability as pure ratios of charges and currents (a la Ampere) or poles. This removes the need for a fourth primary unit for dielectric constant or permeability, and we see here, as in the case of heat, the development which leaves the adherents of the four -dimensional theories not so much to stand on. Along with Lorentz came the Michelson-Morley experiments, disposing of the stationary ether. Then it was Einstein who saw how to retain the body of the classical electrodynamics and the idea of a velocity of light independent of source and absorber, and how to modify the electromagnetic relations for transfer to other reference systems. Dirac’s elucidation of the socalled “electron spin” in terms of relativistic electron waves helps tie in relativity and propagation speed c with electrostatic and magnetic forces. Whether the present relativity is mostly a partial correction for our erroneous as- sumptions in the past, only the future can tell. But the attempts of Page, Milne and others to obtain kinematic relativity, and Milne’s result of an entirely different magnetic force of moving charges, seem to indicate this. To sum up, the speed c, electrostatic forces and magnetic forces are tied up together, whatever system of units one employs, and the basic system tries to make a difficult path a little easier by recognizing this in the simple relation i=q/ct. This leaves c in the units along with its relatives, and not standing as a half-submerged bridge between the units. (1) Warburton, F. W., Am. J. of Physics; 8, 30 (1940). 68 The Kentucky Academy of Science INVERSION APPLIED TO CONICS* SISTER CHARLES Mary Morrison Department of Mathematics, Nazareth College Higher plane curves have a special appeal, and it is interesting to see how such curves are frequently obtained by means of simple inversions. “If two points P, P’ collinear with a given point O are such that the product OP-OP’ is equal to a given constant K, the points P, P’ are said to be inverse with respect to the center of inversion O, K being the constant of inversion. When K is positive, the two points P, P’ lie on the same side of the center of inversion; when K is negative, two inverse points lie on opposite sides of the center of inversion. If the point P describes the figure (F), the figure (F’) described by P’ is said to be the inverse of (F) with respect to the center of inversion” (1). Two important properties of an inversion are immediately evident from the definition; they are: (1). If P approaches the origin, P’ recedes to infinity; and conversely, (2). The points of the circle of radius = \/K, whose center is O are fixed points. This fact is useful in plotting inverse figures, for the points in which a figure cuts this circle will be points of the inverse figure. In this case the point P coincides with its own inverse point. This circle is called the circle of inversion; the constant \/K is referred to as the radius of inversion. If the constant of inversion is negative, there are no points which coincide with their inverse points; that is, the circle of inversion is imaginary. Let us consider first the analytic part of inversion and find the desired _ equations, the two equations involving the coordinates of two corresponding points, P and P’. These equations must express two conditions, that P and P’ lie on a line through the center of inversion, and that OP*OP’=1. Assume K=1. The first of these conditions is satisfied when theA OPM and OP’M’ are similar, for then we may say XG Vie eOk x’ Vi Op’ for the second, if we multiply both numerator and denominator by OP’ we have: OP -OP7OP" 1 1 OP i0p2 | Op? | een Twenty-Seventh and Twenty-Eighth Annual Meetings 69 x y 1 Thus — = — = ————_., and the equations of inversion are f? , 4 4 ee ey x y’ = ete, ; y=. / / x? py Mey Having determined the equations of inversion, let us take a few figures and find the inverse of each. First let us consider the inverse of a straight line which does not pass through the origin. The equation of this straight line is Ax+By+C= 0. Substituting in this equation by the values of x and y in the equations of inversion, we have IN By’ ee + ——— ee xe ty = Dropping the primes we find that this equation may be recognized as that of a circle which passes through the origin. If C =O, in the equation of the straight line, the line passes through the origin; substituting the equations of inversion, ext By’ os ie v BS aie Vie Dropping the primes we have exactly the equation with which we started, showing that the line which passes through the origin is invariant under inversion. Hence, we may make this statement: “The inverse of a straight line which does not pass through the origin is a circle, and a line which passes through the origin is invariant under an inversion.” SEG = Of or Sx by ox Cy? —=20: =O, or Ax’ + By’ =O. Into what type figure will a triangle invert if none of the sides of the triangle go through the center of inversion? Each side will determine a circle, and hence we shall have a figure that is bounded by three curves which are arcs of circles that pass through the center of inversion. (See Figure I.) From what we have seen of inversions we can easily draw the conclusion that the inverse of a system of parallel lines is a system of circles whose centers lie on a line perpendicular to the lines of the system. The equation of the lines parallel to the Y axis is x =a. Substituting ox x x = ———.__ in this equation, _———— =a, or if we drop the primes, 12 , 12 xy” Phy x x? y2 —.__ — 0, which is the equation of circles whose centers lie on the a X axis and which are tangent to each other at the origin. The inverse of a system of lines passing through a point is a system of 70 The Kentucky Academy of Science c ( p fgw Kea, Lt Ariane wavered , awerse, Ko aure, bounded rou fe oy earcles tis, t oy ee of aK Mave rsvaw x aralbo j Weverse , C1s901 ¢ B. % canara Yay evbola ; \Wmver se Wornenustenle \ Bt Ner\ey Cewier oa PANEWSIOM - \ evo a 5 VANGWSe S\yr eons ee oe Cee a 3 NUN EDe Nien CO FO (a < b) A. &Wvkee - coe a 5 pwverse Cardiord (oe Esa ts \raverse Yima cow Ca >b) Ya ¥ Ana daeds Goll) 2 paverser WM evs. Twenty-Seventh and Twenty-Eighth Annual Meetings 7A circles passing through the origin and through the inverse of the point. The family of lines passing through a point of the Y axis has the equation m 1 y = mx-+b. By the equations of inversion x2 y?1 —x——y=0. b b This equation is that of a system of circles passing through the origin and u through (0, —), the inverse of (0, b) through which the lines pass. b Let us next consider the inverse of the circle which does not pass through the origin. The equation of this circle is x?+y?1+Dx+Ey+F—0 where F is not 0. Substituting the equations of inversion in this equation, x/2 we Dx’ Ey’ (0 - Pein) ee (kay)? | (Ray?) (ee) Multiplying by. x+y”, 1+Dx’+Ey’+Fx’+Fy?—0, which is the locus of the circle. If in the general equation of a circle F=0, the circle passes through the origin. When we substitute the equations of inversion in this equation, we get an equation of the first degree. From these results we see “the inverse of a circle is, in general, a circle, but the inverse of a circle which passes through the origin is a straight line.” We might ask this question: “Are there any circles which are invariant under inversion?” Take first the general equation of the circle x’-y?+Dx+Ey+F = 0. We know from what we have just proved that, if F is not equal to 0, the inverse figure will also be a circle, but will this circle be the same circle as the one with which we started? Substituting the equations of inversion in x?--y2+Dx-+Ey+F = 0, Fx’2-Fy?+Dx’+Ey’+1—=0. Notice that, if we let F = 1, we shall get, after dropping the primes, the original equation. Therefore, the condition that a circle be invariant after inversion is that F—1. But what is the condition that_F = 1? F=—1 when the circle is orthogonal to the circle of inversion. EF = 0: If we know the angle of intersection of two circles and take the inverse of these circles, we shall find that the angle of intersection of the inverse circles is the same as the first angle. To do this, we determine the cosine of the angle of intersection of two circles and invert the circles; we find that we get the same value for the cosine of the new angle of intersection. Hence, the angle of intersection of two circles is invariant under inversion. Suppose we invert some conic sections. What types of curves shall we obtain? Take a parabola whose vertex is at the center of inversion. If we choose the origin as the center of inversion, we shall have the equations of the parabola, y? — 2px. Substituting the equations of inversion in this equation, y” x’ (eo L¥7 ae LES ET Ti The Kentucky Academy of Science 1 Multiplying by (x’2+-y’?)?, replacing — by 2a, we obtain x* = y? (2a—x) 2p which we recognize as the equation of the Cissoid of Diocles. (See Figure IIA.) Notice how the curves intersect at the circumference of the circle of in- version. Hence, the inverse points meet, and OP*OP’ = 1. Let us consider the inverse of the aque nea. pete whose center is the center of inversion and whose equation is x2 — y* = a?. Substituting the 1 . equations of inversion, replacing — by a”, we get (x?-+y”)? =a” (x*-y?), 2 a the usual form of the Lemniscate of Bernoulli. (See Figure IIB.) When the center of inversion is at the vertex, we find that the inverse of 1 the equilateral hyperbola whose equation is x?-—— y?-+—x=0 becomes a a+ x ye = <( ) which we recognize as the Strophoid. (See Figure III.) a—x If we take a conic whose focus is at the center of inversion, we shall get a limagon, and the shape of the limacon will vary according as the conic is an ellipse, a parabola, or a hyperbola. The general equation of a conic whose focus is at the origin, or center of inversion, is (1 —e?) x?+-y? — 2e? px — e2p? = 0 where e is the eccentricity of the conic, and x = —p is the equation of the directrix. After substi- tuting the equations in this equation, adding e?x? to both sides, and dividing 1 1 1 by e*p?, we get (x?+y?+ —x)? = — (x*-y?). Now if we set-——=a P e’p P 1 and =b?, then (x?+y?+ax)? = b? (x?+y?). e2p? The locus of this equation is a limacgon and this limacon has the three different forms corresponding to the three forms of conics according as 4 is less than, equal to, or greater than b. When a equals b, we know the limacon is the cardioid. In the parabola e = 1, and x = —1 is the equation =, and ——— —_b*, —=1 =a and ba —1l 1-1 therefore, ab. Thus the parabola inverts into the Cardia the ellipse and the hyperbola give the other forms of the limacon. (Figure IV A, B, C.) It is curious to note that the original curve in each case is rational; that is, the coordinates can be expressed in terms of a parameter and the inverse curve is also rational. If the original curve is irrational, we find that the inverse is also irrational. We could show this by inverting a Cassinian Oval, a quartic curve with only two double points and therefore of deficiency of the directrix; hence, Twenty-Seventh and Twenty-Eighth Annual Meetings 73 one, and we shall obtain a cubic curve with no double point, which is there- fore irrational, since to be rational a curve must have the maximum number of double points which belong to its order. In general, a curve of the n* 1 degree cannot have more than — (n—1) (n—2) double points. An interesting mechanical device known as Peaucellier’s Cell (2) or Inversor gives the mechanical construction of the inverse of a curve. The figure A C, C B, BQ, Q A, P-A, PB is a system of freely jointed rods of mich A €C=B Cand A Q—=OQOB=BP=P A. (Figure V.) At P and Q sockets are placed to carry pencils; a pin fixes C to the drawing board. The system is then movable about C. It is clear that C, Q, P are in a straight line and thatC P*° C Q=C A?—A Q; there- fore, C P-C Q=K, or constant. Hence, whatever curve P is made to trace, Q will trace its inverse, the point C being the pole of inversion. Peaucellier has utilized this construction for the conversion of circular into rectilinear motion. * The author claims no originality for the material of this paper; it has been obtained chiefly from text books on the subject and from seminar papers prepared at the Catholic University of America, Washington, D.C. (1) Altshiller and Court, College Geometry; p. 201 (2) Smith, David Eugene, Source Book in Mathematics; p. 324 THE GENETICS OF INTELLIGENCE EpwarD NEWBURY Department of Psychology University of Kentucky, Lexington, Kentucky If there is included the whole range of ability from feeblemindedness to genius, it may be said that few, if any, psychological subjects have received as extensive consideration from the genetic standpoint as has intelligence. Thorndike’s early conclusion on the basis of the distribution of mental-test performances that there is only one type of human twins has given way to the recognition of monoval and di-oval twins and an extensive research based upon this distinction. The testimony of Estabrook in a 1925 steril- ization litigation that feeblemindedness follows the pattern we identify as that of a Mendelian recessive unit character has been controverted by later evidence showing that at least such feeblemindedness is not the only familial type, and may be rare if existent at all. There is an increased knowledge of various environmental factors such as birth injury and thyroid insuf- ficiency which may contribute to feebleminded development. There is a recognition that both heredity and environment influence the development of intelligence. This conclusion has been supported by evidence, especially from twins and siblings, although it might have been reached theoretically 74 The Kentucky Academy of Science from the basic assumption now generally accepted that both heredity and environment are necessary factors in development. This basic assumption and its implications in methodology and formulation of the problem have been among the more revolutionary contributions in the last few years. Indeed, the comparative studies of twins and siblings, raised under similar and different environments, may be traceable to this theoretical reorienta- tion. It is with an extension of such considerations that we shall be con- cerned today, notably as they apply to the problems of the limiting influence of hereditary and environmental factors and the nature of intelligence. If we accept a dynamic view of development, then in the organism at the stage at which we can be concerned with its intelligence, any behavior is a function of its heredity and its environment, both of which are necessary factors in the development of that behavior. This is a viewpoint ably pre- sented by Woodworth and Jennings, extended by others, and now receiving wide acceptance in textbooks. In the dynamics of behavior, however, the developmental factors alone are insufficient to account for the behavior, so that it is necessary to include the immediate factors producing the be- havior, the stimulating situation. When a stimulus itself may be said to be internal, we must still include the external medium in which the individual is Operating at the moment. We must say then that behavior is a function of heredity and environment and situation. This situation may be regarded as quite as essential in the study of intelligence, as perhaps in every other case, as any of the developmental factors. The test itself and the testing situation comprise this last factor in the dynamic arrangement from which the behavior results. For the present we may side-step the issue of whether the behavior is a function of the individual and his environment taken as a single configuration or whether it is a function of the individual in a given environment. In either case, the situation is a necessary contributor to the behavior, as thoroughly essential as either of the two developmental fac- tors. In such a dynamic arrangement, in order to study the influence of any one factor, it is necessary to hold constant the other two factors, or else to produce similar and simultaneous changes in both an experimental and control group, leaving only the third factor to be changed or to be varied between the two groups of subjects. Here is a basic methodological consideration which cannot be escaped if one accepts the dynamic view of development and stimulation of behavior. It may be for this reason that family history studies, which were once importantly displayed in textbooks, are now receiving less emphasis. As studies in the dynamics of family life, from the sociological point of view, they may well receive more emphasis than heretofore, but from the stand- point of genetic analysis, they can only be used when it. is plain that so far as the trait in question is concerned, the environmental variations from locality to locality and from generation to generation are ineffective in influencing the trait. Familial feeblemindedness, then, is familial feeble- mindedness; the variant is not necessarily hereditary. In studies on twins and siblings and foster children, efforts. have been made to take into account the operation of the two sets of developmental Twenty-Seventh and Twenty-Eighth Annual Meetings 75 variables. As already pointed out, a demonstration that intelligence is subject to both hereditary and environmental variations supports a con- clusion which could have been reached theoretically on other grounds, in- deed, arises as necessity from the dynamic view of development. Answers to two further questions, both related, have been sought, however, in this work: namely, the limits of the influence of either the hereditary or the environmental factors, and the relative importance of the two. So far as the development of the individual is concerned, since both heredity and en- vironment are necessary factors, it is not possible to attribute a relative importance to either one or the other set of factors, for contributing factors which are essential are neither more nor less important than any other es- sential contributing factors. Consequently any statement, based on such work as that of Burks, that the contribution of heredity is 75 or 80 per cent., that of environment 20 or 25 per cent., is not tenable on theoretical grounds—a fallacy recognized by that investigator. On the other hand such statements may have some significance in de- scribing not individual development but the factors effective in producing variation from individual to individual within a given population in a given community. At best the relative effectiveness of the fluctuations in either variable, heredity or environment, would hold, as emphasized by Hogben and by Schwesinger, only for the particular heredities available in such a popu- lation and for the particular environments found in such a community. As these investigators have pointed out, in a human society the relative im- portance may shift with conditions varying with time. A study such as that of Burks, concerned with foster children, can hardly be taken as typical of the United States on either basis. The study is limited further by the time factor, is conditional upon the time it was made. As we learn more and more medically and pedagogically about the development of children it may be expected that the environmental variant will be increased. Furthermore, as medical and _ psychological knowledge reaches the communities more extensively and effectively, we may expect a still further increase in the environmental variant. In a cretinous group, for example, the parental variant (which is really what Miss Burks was getting at rather than heredity) would be markedly reduced in importance with the introduction of thyroid medication. This view is supported by the Chicago researches on identical twins. Their investi- gators conclude that the hereditary variant is relatively reduced when the twins are raised apart. This conclusion can be reached theoretically without reference to these studies. These limitations are necessary even though the measure of relative im- portance is a valid one. Although the point need not be pressed here, there are grounds for concluding, after Hogben, that such a measure cannot be made under natural conditions. The changes produced by varying the environment depend upon the heredity on which the environment is oper- ating. The changes produced by varying the heredity depend upon the environment in which the heredity is developing. Under experimental conditions one would deal with a uniform stock and vary the environment; 76 The Kentucky Academy of Science then change to another uniform stock and vary the environment, similarly. The maximum or average effective variation in the two cases would not necessarily be the same. In studying the hereditary variant the conditions would be reversed. The method of partial correlations cannot be a sub- stitute for such a control of experimental conditions. In a conglomerate stock, with a multifarious environment, it is difficult to see how statistical manipulation can be a substitute for holding the stock or the environment constant. And under experimental conditions, with a limited number of environmental variations, say, five, and a limited number of changes in stock, say, five, in a situation where it would be possible to try all five environmental variations with each heredity, and all five hereditary vari- ations with each environment, it is a fair question to ask just how one would conclude regarding the relative importance of each variable in pro- ducing changes. It is a question then whether statements regarding the relative importance of the variables in producing changes in a given popu- lation are causative statements at all; they may only be statistical descrip- tions of the relationships between savailes i the population and nothing more. Hogben has shown that, whatever the validity of statistical measures of relative importance, they do not, for reasons indicated in the preceding paragraph, determine the limits of change which might be pro- duced environmentally. The question of the limits set by heredity or environment is itself an important one. One familiar position is that of Snyder, who holds the evidence to indicate that ‘“‘each person has an innate mental ability beyond which he is incapable of developing. The upper limit of this ability is determined by hereditary factors.” This is a common position. It im- mediately raises the perplexity, however, of what to do with cases such as cretinism, where the limits in some cases at least are clearly set by the nutritional environment. On theoretical grounds, the position is open to the objection that among two necessary factors for producing a result, each sets the limit of variation in the result. If a maximum intelligence performance might be obtained, both hereditary and environmental factors would contribute to it and if either were changed effectively, as we know they can be, the maximum would no longer be obtained. Hence both are limiting factors and to the same degree. On this basis, then, studies on the limits of change, such as those of foster children and identical twins reared apart, have significance only as applying to the particular population in question in the particular environment in question and at a particular time. One may conclude that among foster children the I.Q. can be raised 4-7 points on the average by placing the children in superior homes; that among identical twins reared apart different environments can change the I.Q. on the average 8 points. These statements cannot be regarded, how- ever, as generalizations applicable to all other communities and groups nor as limits of the influence of varying the environment, because they apply only to specific groups and conditions and are subject to all the limiting con- siderations just described. In the Chicago study on identical twins reared apart, for example, while the average difference in I.Q. was 8 points, in a Twenty-Seventh and Twenty-Eighth Annual Meetings Wi. third of the cases the differences were as great as 23 points in M.A. and as much as 12 I.Q. points. Differences in mental age ran as high as 46 months and in I.Q. as high as 24. If these twins were identical, why were some changes so much greater than others, which were as little as 1 month and 0 I.Q. points? From the standpoint of the dynamics of development, this seems to be the important question. If community studies are made, applicable only to certain communities, it is necessary to identify the char- acteristics of the communities so that they may be identified as similar to other communities where the same situation might hold, as for example that certain changes in orphan I.Q.’s might be expected. If on the other hand the interest is in the process of development, it is specific agents which can be effective variables which must be identified. Since the limits of change in the development of intelligence are functions of both heredity and en- vironment, the discovery of the ultimate limits must await our combination of all possible heredities with all possible environments. Meanwhile the problem would be one of what specific factors could produce what specific changes in development. The mere collection of cases of identical twins reared together or apart with a striking of average differences will not furnish light on this problem. One need not study identical twins to obtain great differences in intelli- gence records, for these may be obtained from the same individuals. Data compiled by Prof. G. B. Dimmick from the records of the Child Guidance Service at the University of Kentucky show, out of a total of 414 cases differing at least 2 LQ. points in 1937 Stanford-Binet and Arthur point scale examinations, 33 cases in which the point scale I. Q. is 20 points or more higher than the Binet, and 11 cases in which the Binet is as many points high- er than the point scale. This difference clearly changes the individual’s classification, which in some cases is from feebleminded to normal, or vice versa. These data are striking illustration of facts which have accumulated since the World War from the comparison of different language groups, negroes and whites, males and females, rural versus city groups, and from other sources, showing that the general intellectual level and indeed in many _ cases the J.Q. obtained is a function of the type of test administered. These facts have been noted by Anastasi, who has also emphasized the accumulation of evidence which, taken with the nature of test construction, shows that test performance may be a function of the cultural framework within which a given test has been constructed or is applied. The inescapable question is raised as to what is meant by intelligence. In the dynamic analysis of behavior not only was it indicated that the develop- mental factors of heredity and environment were necessary contributors, but it was also pointed out that the immediate situation must be included as an essential factor in the analysis. In the case of intelligence the difficulty is that the behavior which is actually immediately measured is not of con- cern, but a measurement is inferred from that behavior. The difference between an achievement test and an ability test lies in the assumptions back of the test items, in the latter case it being assumed that a common environ- ment has been available to all those tested. Now when the developmental 78 The Kentucky Academy of Science environment is modified, is the modification of intelligence or of the achievement? It appears that intelligence represents some sort of poten- tiality. To assume that it is an innate faculty as is sometimes done, pre- sumes that no language development is necessary. If intelligence represents purely some maturational organization of the individual, the dynamics of development will be of a different sort than if one presumes that schooling and other experiences do have some effect in developing intelligence. There is no real assurance that various intelligence tests are measuring enough of the same factor in performance as to preclude wide differences between in- dividuals on the basis of experience, even though general environment and schooling be the same. Specific tests depend upon behavior which may or may not require some kind of environmental contribution to develop- ment—that is with the usual stock. Language knowledge is only the most obvious instance. Therefore, the validity of a given intelligence test in a given practical situation is no assurance that it will be a valid measuring instrument for an inferred potentiality or organization in a developmental study where deliberately the various factors in development have been changed or are allowed to change. The differences between various test results on the same individuals, either with different types of tests or with some time lapse between testing, are frequently as great as the differences between identical twins reared apart. They are large enough, then, that in developmental studies they cannot be ignored. Some standard of intel- lectual measurement not susceptible to developmental factors which can be included or explained away depending upon one’s definition of intelligence is necessary at the present time unless the further demonstration of develop- mental influences on test behavior is to produce confusion rather than an increase in the knowledge of how factors in development influence in- telligence. One further point. It is customary now to refer to the normal distri- bution curve of intelligence records obtained on various tests and to infer that the genetic factors operating in this case must be numerous. An as- sumption of two sets of cumulative factors will yield, however, five quantitative gradations in a given trait. The assumption of three sets will yield seven such gradations. When one considers the variations in test re- sults which might be produced by a wide variety of environmental var- iations as well as by unreliabilities in the testing itself, it appears that in order to duplicate the ordinary distribution curve for intelligence the number of factors would certainly not have to be more than three allelo- morphic pairs. Environmental influences should easily smooth out the demarcations between these seven gradations. It is possible that even two such pairs would suffice. REFERENCES Anastasi, Anne. Differential Psychology: Individual and Group Dif- ferences in Behavior. Pp. 615; Macmillan. New York. 1937. Burks, B.S. The relative influence of nature and nurture upon mental development; a comparative study of foster parent-foster child re- Twenty-Seventh and Twenty-Eighth Annual Meetings 72 semblance and true parent-true child resemblance. 27th Yearbook, Nat. Soc. Stud. Educ. Part I; 219-316; 1928. Hogben, Lancelot. Nature and Nurture. Pp. 144; Norton. New York. 1933. Jennings, H. S. The Biological Basis of Human Nature. Pp. 384; Norton. New York. 1930. Laughlin, H. H. The legal status of eugenical sterilization. Supplement to the Annual Report of the Municipal Court of Chicago for the Year 1929. Pp. 83. Newman, H. H., Freeman, F. N., and Holzinger, K. J. Twins: A Study of Heredity and Environment. Pp. 369; Univ. Chicago Press. Chicago. O37. Schwesinger, G. C. Heredity and Environment: Studies in the Genesis of Psychological Characteristics. Pp. 484; Macmillan. New York. OB: Snyder, L. H. The Principles of Heredity. Pp. 385; ends. New Work, 193:5:. Thorndike, E. L.. Measurements of Twins. Arch. Phil., Psychol. and Sc. Meth. No. 1; Pp. 64; 1905. Woodworth, R. S. Psychology (3rd ed.) Pp. 546; Holt. New York. 1934. HEMOLYTIC STREPTOCOCCI FROM THE THROATS AND TONSILS OF PATIENTS WITH CHRONIC TONSILLITIS (abstract) By Doris McKenzie, R. H. WeEaveER, anv M. SCHERAGO Department of Bacteriology, University of Kentucky Attempts were made to isolate hemolytic streptococci from the throats of 42 patients before the removal of their tonsils and from the surfaces and interiors of the excised tonsils. All of the strains that were isolated were classified serologically (Lancefield groups) and their fibrinolytic properties, final hydrogen ion concentration and ability to hydrolyze sodium hippurate and to ferment trehalose, sorbitol, esculin, xylose, mannitol, arabinose, salicin and lactose were determined. The results of the pathologist’s ex- amination of the tonsils were available for comparison with the bacteriolog- ical findings. Seventy-three cultures of hemolytic streptococci were obtained from 35 (83%) of the patients. Of the 73 cultures 47 (64%) belonged to group A, 14 (19%) to group B, 3 (4%) belonged to group Cc and g (13%) elomecd to groups other than A, B and C. The cultures which were isolated were very diverse biochemically, e.g. the 47 cultures of group A represented 23 distinct strains. In very few cases were the same strains isolated from the interiors of the tonsils that were found on the surfaces. No correlation was found between the bac- teriological and the pathological findings. 5 } A comparison of these results with those of earlier investigators who have studied hemolytic streptococci from the throats of normal individuals — shows the following differences. 1. Hemolytic streptococci were isolated from a larger percentage of these patients than from normal individuals. 2. A larger percentage of Group B strains were isolated. Davis and Guzdar found no group B strains in the throats of Hong Kong Chinese and Hare found that 5% of the strains isolated from normal throats in England belonged to group B. The large percentage of group B strains encountered in this study is surprising in view of the rare occurrence of group B strains in human infections. 80 The Kentucky Academy of Science ANATOMY OF THE ALIMENTARY CANAL OF BRUCHUS QUADRIMACULATUS (COLEOPTERA) WITH SPECIAL REFERENCE TO THE HISTOCYTOLOGY OF THE MID-GUT EPITHELIUM IN THE NORMAL (UN-FED) AND FED CONDITIONS (abstract) Epcar P. JAYNE Department of Zoology, University of Kentucky The alimentary canal of Bruchus consist of three principal divisions: fore-gut, mid-gut, and hind-gut. The tissue layers from inside outward are as follows: an epithelial layer, a circular muscle layer, and a longitudinal muscle layer. Although the adult beetle normally does not feed, it was found that it could be fed, thus increasing its normal span of life. In the normal (un-fed) condition of the mid-intestinal epithelium, there is con- siderable sloughing off of the tissue. The sloughing off of the nucleo- plasmic and cytoplasmic tissues may be interpreted as a mechanism whereby the animal is nourished in the absence of other sources of food. While some of the cells are sacrificed for food, others elaborate secretion by the apocrine method. On the other hand, the mid-intestinal epithelium of beetles which have been fed show little sloughing off, and the food is digested by secretion formed by the merocrine method. In both the fed and un-fed conditions the epithelium is rejunivated by nuclear replenishment which have their origin in the nidi. The degenerate nuclei of the epithelium probably act as catalysts which in turn are responsible for the breaking down of the epithelium. It is suggested that the mitochondria are con- cerned either directly, or indirectly with the elaboration of secretion globules. AKT TRANSACTIONS OF ‘THE KENTUCKY ACADEMY OF SCIENCE (Quarterly Series) AFFILIATED WITH THE A.A.A.S. VOLUME 9—NUMBER 4 MARCH 1942 TWENTY-SEVENTH AND TWENTY-EIGHTH ANNUAL MEETINGS / 1940-1941 rz ie CONTENTS Page The Advantages of the Spectrophotometer to Increase the Accuracy of Denigés’ Method for Determining Phosphorus and Arsenic INianeitiecE:® ON ec lesaieket | os Wii PE oy ek NS on CR 81 Letters to Robert Peter VERE Ray ney Ge ater ee anes re 5 0 9c Pi) ok. "Sat oe eR ACL 88 Relation between Frogeye and Greenspot of Tobacco Stephen Diachun W. D. Valleau Be Nie olitisone:, Semper ies sce. cl kOe eis tg Salah ther sea Rae 90 Nutritional Problems in the Production of ‘Thoroughbred Horses (abstract) Mw PN oglachetee 2. 2.5 sete thapss be r3 Pt e etee eH t ete ae eee 92 The Use of Corn Gluten in Iodine Deficient Diets. A Preliminary Report (abstract) Wee Si Ed tassel ches ua et Bees tala 4M ai: 93 RE MITGM ING Wise een ne it ais Mean eer ae ean ANS. 8 SR 94 us. Selec ye 90 9 SNR noes Rear ie eo a cement? Oe 95 TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE EDITORIAL STAFF John Kuiper ......... Lexington, Kentucky .. Editor-in-Chief Charles Barkenbus ....... Lexington, Kentucky .......... Chemistry pee Blache: it Jinan mas Morehead, Kentucky ............. Physics EUW Cooks a Aa 8 or ee Ho Danville, Kentucky .......... Bacteriology W.R. Hutcherson ........ Berea, Kentucky ............ Mathematics Jobn ™ Kuiper’: 35,0) oan" Lexington, Kentucky ....... Psychology & Philosophy HB Lovell 0.040 ee Louisville, Kentucky, ))). 4/7 Biology Ae OP oNicharlane #70) 2 Lexington, Kentucky ............ Geology Manuscripts. The Transactions must be limited to the proceedings of the annual meetings of the Kentucky Academy of Science and to original manuscripts pertaining to science. Under present financial limitations each article must be limited to approximately five pages of the Transactions. Manuscripts are subject to the approval of the Editorial Staff. Manuscript material may be submitted to the Associate Editor of the subject covered or to the Editor-in-Chief. Extra-Cost Features. ‘The extra cost of special features such as cuts, graphs, tables, etc. above the text-run price of $2.65 per page must be borne by the contributor. The Editorial Staff will advise contributors con- cerning the extra cost of special features upon receipt of manuscript. Illustrations to be included in an article should accompany the manuscript if possible, or, if sent in separate package should be properly labeled as to the article in which they are to occur. - Proor. Galley proof will be sent for approval of contributors. The proof should be returned promptly to the Berea College Press, Berea, Kentucky. REPRINTS. Reprints are furnished at publisher’s prices by negotiating directly with Berea College Press. Price quotations on reprints are sub- mitted with the proof. Orders for reprints should accompany the proof to the Berea College Press, Berea, Kentucky. SUBSCRIPTION RaTEs. The Transactions is sent without additional ex- pense to all members of the Kentucky Academy of Science who are not in arrears for annual dues. The annual subscription rate’ for non-members is $2.00 in the United States and Canada, $2.50 in foreign countries; single number 75 cents. One volume of four numbers appears each Academy fiscal year. BusINEss CORRESPONDENCE. Remittances and correspondence concern- ing subscriptions, extra costs, and other financial matters except reprints should be addressed to John Kuiper, University of Kentucky, Lexington, Kentucky. aoe Twenty-Seventh and Twenty-Eighth Annual Meetings 81 THE ADVANTAGES OF THE SPECTROPHOTOMETER FOR MEASURING THE INTENSITY OF THE BLUE COLOR IN DENIGES’ METHOD FOR PHOSPHORUS AND ARSENIC Martin E. WEEKS Agricultural Experiment Station University of Kentucky The precise determination of small quantities of phosphorus in natural materials such as soil extracts, biological materials and mineral waters has been aided greatly by the development of the various colorimetric methods of estimation. One of the most sensitive and accurate of these is the Denigés method as modified by Truog and Meyer (2). The greatest disadvantage of this method is its sensitivity to slight variations in amounts of the various reagents and acid concentrations and to the effect of certain other ions in altering the intensity and quality of the color. The effect of certain of these factors has been studied more fully by Chapman (1). After using this method for several years, the author has found it to have certain limitations when colors were compared in Nessler tubes and that great care is necessary in preparing standard and unknown solutions so that the greatest precision can be obtained. When the colors are compared in Nessler tubes, the accuracy of readings in the optimum range can be held to within about plus or minus two percent. If, however, the intensity of color in the unknown varies more than about thirty percent from the standard and particularly if a small amount of ferric iron or organic ma- terial is present, greater errors may be expected. In very dilute solutions it is often difficult to get good results in determining phosphorus in soil ex- tracts when very dilute standards are used because of off-color tints in the unknown. With the aid of a spectrophotometer, it was possible to investigate some of these factors and discover their effect on the determination. It became apparent that with the use of an instrument of this type where a standard phosphorus curve is constructed it is necessary to observe certain precautions that have far less effect when standards are prepared for each set of de- terminations. It is the purpose of this communication, then, to evaluate the difficulties inherent in the ordinary colorimetric determination and to point out the precautions to be observed in the electrometric method. 82 The Kentucky Academy of Science The Use of the Spectrophotometer The spectrophotometer used in this study was a Coleman Model 10 Double Monochromator Spectrophotometer. The percentage transmission was measured by means of a Coleman Model 3-D pH meter, used as a po- tentiometer.* In selecting the best spectral region for the determination of phosphorus it was necessary to measure the percentage transmission of a standard phos- phate solution at various approximate wave lengths. As ordinarily used, the instrument passed a beam of light on to the cell thru a slit with an opening of about thirty millimicrons. These measurements were made at intervals of thirty millimicrons or less between 370 and 980 millimicrons, on standard solutions at three different acid concentrations and with slightly different amounts of stannous chloride. The data for representative values of these measurements are shown in Table 1. Table 1. Percentage transmittance of a standard phosphate solution at various wave lengths. Phosphorus concentration—0.25 mgms. per liter. Acid concentration—.478 N. SnCls 7 drops per 100 ml. Wave Percent Wave Percent Wave No. length transm. Wave No. length transm. 0.2 370 75.8 Wiloll 690 45.2 1.0 380 78.2 11.2 700 45-2 2 410 82.6 12.0 710 45.7 3 440 83.6 12.1 720 46.0 4 470 81.7 WA 730 47.0 5 500 78.6 13 740 48.8 6 530 74.2 14 770 53.6 7 560 67.2 1D) 800 57.6 8 590 59.0 16 830 52.7 5 620 52.0 7 860 63.0 10 650 48.3 18 890 67.5 10.1 660 47.6 19 920 71.8 10.2 670 46.6 20 950 76.7 11.0 680 45.7 21 980 80.5 The data presented are for a phosphorus standard solution made up in .478 normal sulfuric acid, which is a higher concentration than that recom- mended by Truog and Meyer (2). The only effect the acid concentration (between .35 and .5 normal) had on the transmission values of a pure *Both the Model 10 D. M. Spectrophotometer and the Model 3-D Elec- trometer were provided thru the courtesy of the Cincinnati Scientific Company, 224 Main Street, Cincinnati, Ohio. i = Twenty-Seventh and Twenty-Eighth Annual Meetings 83 standard phosphate solution was to raise or lower them slightly. It can readily be seen from these data that maximum light absorption is obtained in the region between 680 and 700 millimicrons and that for quantitative comparison except for special purposes the measurement of the blue phosphorus color should be made in this region. However, unless other- wise stated in this work, all measurements were made at 650 millimicrons because it seemed that at this wave length the color reaction was affected least by interfering ions such as ferric iron. Phosphorus concentration curves were drawn at three different acid con- centrations, and at several wave lengths at points near the bottom of the curve between spectral region Nos. 8 and 14 (See Table 2 and Figure 1). The curves were fairly smooth and all passed thru the origin, thus showing that the reaction was proportional to concentration. It was found that when the concentration of phosphorus was higher than .6 to .7 milligrams per liter, the concentration curve became so steep that good comparisons were quite difficult to obtain. Table 2. Spectral transmittance of standard solutions of different con- centration. Percent No. Conc. p.p.m. Wave No. Wave Length Transmission 1 .05 8 590 87.6 10 650 83.9 12 710 83.0 ‘ 14 770 85.8 2 .10 8 590 78.3 10 650 71.3 12 710 71.0 14 770 76.8 3 15 8 590 70.4 10 650 62.3 12 710 60.6 14 770 67.8 4 .20 8 590 63.0 10 650 53.5 12 710 51.8 14 770 58.2 5 ey) 8 590 57.1 10 650 46.7 12 710 43.6 14 770 52.6 6 30 8 590 ») Pa nl 10 650 40.8 12 710 37.0 14 770 48.0 84 The Kentucky Academy of Science Effect of Acid Concentration on the Development of Color. As the acidity of the solution in which the color is being developed in- creases, the less intense is the resulting color. ‘This relationship is shown in Table 3. In acid concentrations below .35 normal, the color is very intense due to a reaction between the excess molybdate and the stannous chloride reagents. In acid concentrations between .35 and .5 normal the color is due only to the reduction of the ammonium phosphomolybdate molecule and the color is proportional to the phosphate concentration. In this range, the color is a uniform light blue that is very easily compared by the eye, while at lower concentrations the color has a purplish tinge that is more difficult of comparison by visual means. Table 3. Effect of acid concentration on light transmission. Phos- phorus conc.—.25 mgm. per liter. Acid conc. Percent Acid conc. Percent normality. transmission. normality. transmission. 16 72 46 47.6 .20 16.5 48 48.6 24 2E6 50 48.6 .28 30.2 54 T10 30 33.6 58 54.4 24; 38.2 62 59.5 38 42.0 -66 63.0 40 43.5 70 73.0 42 44.9 Ta 81.0 The time required for full development of the blue color varies directly with acid concentration. It was proposed by Truog and Meyer that the determination be carried out in .4 normal sulfuric acid solutions because at this concentration the full color would be developed within one minute. It was found, however, that this is not strictly true, but that the time varies between 3 and 10 minutes. The following table shows the time required for full color development in standard solutions at room temperature. Acid concentration Time in normality. minutes. 34 3 40 5 44 5 48 | 10 For practical purposes, when comparisons are made in Nessler tubes or colorimeters and when the acid concentration is below .44 normal, readings can be made in one to two minutes without seriously affecting the accuracy of results because the change after this period is usually within two percent. 85 Twenty-Seventh and Twenty-Eighth Annual Meetings 025 °20 e Vv 1 10 ro) uo Concentration Mgms.per liter. 100 80 Figure I 70 60 50 Percent Transmission ho 30 20 l. “Wave No. 8 or 590 millimicrons Wave No. 10 or 650 u Wave No. 12 or 710 ! Wave No. 14 or 770 " 10 86 The Kentucky Academy of Science Effect of Stannous Chloride on the Development of Color. One of the greatest difficulties encountered with Denigés’ method for phosphorus is the instability of the stannous chloride reducing agent. It was formerly necessary to make this solution up every day or two, but Truog and Meyer found that oxidation could be retarded by covering the solution with a thick layer of white mineral oil. So protected, the solution keeps fairly well for several months, although it loses strength continually. This factor has some importance, because a variation in the amount of reducing agent affects the intensity of the color as shown in the following table where a standard phosphate solution was reduced with a varying amount of freshly prepared stannous chloride. (The amount of reducing agent was measured in all cases by counting drops.) Drops SnCl, Percent Transmission 2 51.5 4 50.0 5 48.0 6 47.6 7. Ais) 9 46.7 11 45.5 1S 43.8 20 42.8 25s 42.3 On the average, one drop of stannous chloride in amounts between 5 and 13 drops changed the percent transmission of light by .5 percent. The amount of stannous chloride added had little effect on the time re- quired for full color development or on the time before the color began to fade. It was found in most cases that after the color was fully developed, it remained practically constant for a period of about ten to twelve minutes and then very suddenly began to fade. The addition of another drop of stannous chloride would bring it back to its original intensity for another ten-minute period. This process could apparently be repeated several times even after the standard had been made up for several hours. Effect of Ferric Iron on Color Development. The most common interfering ion that affects the formation of the blue color is ferric iron. The presence of more than about one to two parts per million of this ion causes a greenish tinge and a decrease in intensity that prevents accurate matching in an ordinary colorimeter. An experiment was conducted to see what effect varying amounts of this ion would have on the transmission of light in the spectrophotometer. The results are shown in Table 4. Standard phosphate solutions were treated with varying Twenty-Seventh and Twenty-Eighth Annual Meetings 87 amounts of iron in the form of ferric chloride or ferric ammonium sulfate. Table 4. Effect of the ferric ion on development of the blue color in standard phosphate solutions. Spectro pho- Form of p.p.m. tometer Nessler tube Color of iron. Fe +++ readings readings solution. FeCl; 0 49.9 50 blue 10 5053 54 greenish 20 49.8 Wi i 30 49.2 62 iid 40 48.8 62 - FeNH, 5 50.5 53 blue (SOx) 2 10 51.0 55 greenish 20 oS 61 ” The results indicate that when the spectrophotometer is used ferric iron has much less effect on the accuracy of the results. The presence of 10 to 20 parts per million changes the value of the readings by about 1 percent or less, while this amount of iron makes it impossible to obtain a comparison with a standard in the colorimeter or Nessler tube. Colorimetric vs. Photometric Methods of Comparison. By the use of the spectrophotometer the precision of comparisons can be increased to the point where a group of standards prepared from day to day can be read with a maximum error of about 0.5 percent. However, unless certain precautions are observed, the accuracy may be no greater than where comparisons are made in Nessler tubes and a standard is prepared for each set of determinations. When a spectrophotometer or similar instrument is used a standard curve is drawn and unknowns are compared with this curve. The accuracy of the determination then is dependent upon how closely conditions are duplicated in the preparation of the unknowns as compared with the standards from which the curve was constructed. For best results, the sulfuric acid am- monium molybdate should be prepared by titrating to the same value each time it is prepared and this reagent should be delivered to the sample with a good burette. The stannous chloride reagent should be freshly prepared at frequent intervals and protected by a half inch layer of mineral oil. Some method should be provided whereby this reagent can be measured into each sample with a fair degree of accuracy Finally, the standard curve should be checked from time to time with a standard solution to see that conditions have not changed. As stated above, these precautions are not as important when comparisons are made by ordinary colorimetry both because in this 88 The Kentucky Academy of Science case a standard is prepared each time a set of unknowns is to be run and because the accuracy of comparison is much lower. If any appreciable quantity of ferric iron is present the use of the spectrophotometer gives a far greater degree of accuracy than the usual colorimetric methods. Bibliography 1. Chapman, H. D. Notes on the use of glass color standards for de- termination of phosphorus by Denigés’ colorimetric method. Ind. and Eng. Chem.Anal. Ed.3: 282-4, 1931. 2. Truog, E. and Meyer, A. H. Improvements in the Denigés colorimetric method for phosphorus and arsenic. Ind. and Eng. Chem. Anal. Ed. We SS Wee). LETTERS. TO ROBERT PETER V. F. Payne Department of Chemistry Transylvania College, Lexington, Kentucky The professional letters addressed to Dr. Robert Peter during the years from 1828 until the time of his death in 1894 have been secured by Transylvania College from his family and are a part of the Robert Peter Collection in the Transylvania Library. In the collection there are also lecture notes made by him in the classes of Rensselaer and Transylvania, notes on and complete class and public lectures of his own, scrap books, and original manuscripts of various writings. The carefully preserved letters antedate his arrival in Lexington in 1832 but are most abundant for the great period of activity of Dr. Peter in Lexington. ‘The letters, approximately 870 in number, have come from about 385 writers. The letters have come primarily from points in Ken- tucky and other central, eastern and southern states but occasional letters came from abroad. ‘The letters arise from the mutual interests of Dr. Peter and his correspondents in the applications of chemistry, medicine and other sciences. Many writers of letters to Dr. Peter were typical seekers for the counsel of a learned professional man, chemist and physician. Typical of the scientists and professional men who have written Dr. Peter are: Elisha Bartlett (1805-55), professor in the Medical Department of Transylvania, Maryland, Louisville, College of Physicians and Surgeons of New York City, fifteen letters; John Lee Comstock (1789-1858), physician and author of science books, nine letters; Twenty-Seventh and Twenty-Eighth Annual Meetings 89 James Dwight Dana (1813-95), geologist, zoologist, one time editor of Silliman’s Journal, two letters; Elias Durand (1794-1873), French botanist, member of the medical corps of Napoleon’s Army, nine letters; Amos Eaton’ (1776-1842), famous science teacher and author, two letters; Joseph Henry (1797-1878), famous physicist, one letter; Samuel Prescott Hildreth (1783-1863), physician, naturalist, author, twenty-four letters; Oliver Wendell Holmes (1809-94), writer, physician, teacher, three letters; Thomas Sterry Hunt (1826-92), eastern chemist, geologist, writer, one letter; Leonidus Moreau Lawson (1812-64), physician, graduate of and professor in Transylvania University, also professor in the Ohio Medical College, Kentucky School of Medicine and the University of Louisiana, author, sixteen letters; Leo Lesquereaux (1806-89), Swiss scientist who came to the United States in 1848, worked with Agassiz, lived in Cambridge, Mass. and Columbus, Ohio, ten letters; John William Mallit (1832-1912), southern chemist and geologist, coming to the United States from Ireland, three letters; Thomas Duche Mitchell (1791-1865), physician, chemist, author, teacher in the Medical Department of Transylvania University and elsewhere, three letters; David Dale Owen (1807-60), chemist and geologist for several state geological surveys, seventy-five letters; Richard Owen (1810-90), geologist, brother of David Dale Owen, twenty-two letters; Benjamin Silliman (1779-1864), famous chemistry and natural science teacher of Yale, founder of the American Journal of Science (Silliman’s Journal), one letter*; Benjamin Silliman Jr. (1816-85), successor to his father, six letters*; John Lawrence Smith (1818-83), famous chemist, long a resident of Louisville, eight letters* ; Lunsford Pitts Yandell (1805-78), professor of chemistry in Transylvania University and later in the University of Louisville, two letters. Transylvania College invites study of this collection and hopes to add to it duplicated copies of Dr. Peter’s letters in exchange for duplicated copies of the letters addressed to him. *Letters to Robert Peter. JI. From J. Lawrence Smith, II. From B. Silliman and B. Silliman Jr., read before the Division of the History of Chemistry of the American Chemical Society in St. Louis, April 9, 1941. 90 The Kentucky Academy of Science RELATION BETWEEN FROGEYE AND GREENSPOT OF TOBACCO STEPHEN DiacHuNn, W. D. VaLLEAU, AND E. M. JOHNSON Agricultural Experiment Station University of Kentucky Within the last few years the frogeye leafspot disease of tobacco has become prevalent in Kentucky. Simultaneously a greenspot of cured Burley tobacco has become rather common. It has been suggested(1) that frogeye and greenspot are caused by the same fungus, i. e., greenspots are late frog- eye infections which are initiated in the field a short time before cutting and develop during curing. Most of the investigation on these diseases has been carried on in foreign countries, principally Australia(2), Rhodesia, Sumatra, and Ceylon. Very little work has been reported in this country, and there seems to be no positive proof of the identity of the causal pathogen of these two diseases on Burley tobacco. Because of this state of uncertainty, definite information about the relation between the two types of spots seems desirable. There- fore, the following study was made. Description of the diseases. Frogeye spots develop on the leaves of growing plants in the field, and persist on the cured leaf. The spots are 4% to 4 of an inch in diameter, usually circular, and brown, frequently with a gray or white parchment-like area in the center of the spot where dark, tufted conidiophores are produced. The spots. are surrounded by an indistinct yellow zone which blends into the normal green of the leaf. Under some conditions the center of the spot does not bleach, but remains brown. The spot may then be zonate, and increase to a diameter of 1% inch or more, and may not produce spores. The frogeye disease is caused by the fungus Cercospora nicotianae. Greenspot occurs only on cured tobacco and is usually confined to leaves on the upper half of the plant. The greenspots are 1/16 to ™% inch in diameter, round or angular, and may be sharply defined or may blend grad- ually into the surrounding healthy tissue. The color is variable: the spots may be green on both surfaces of the leaf, green on the upper and gray on the lower, or blue-green, gray-green or nearly black on both surfaces. Spores are not present on the spots. Comparison of fungi isolated from frogeye and greenspot. Isolations were made from frogeye and greenspots. Spots were cut from leaves, washed in Twenty-Seventh and Twenty-Eighth Annual Meetings ot 1/1000 HgCls 15 seconds, rinsed in 4 changes of sterile distilled water, and plated on potato-dextrose agar. Cultures were then made from the fungi growing out from the spots onto the agar. Isolations were made from 97 leaves, each leaf taken from a crop produced by a different grower. The leaves were obtained from the 1938 and 1939 crops. A total of 423 green- spots and 117 frogeye spots was cultured. Of these, 405 greenspots and 110 frogeye spots yielded fungi which were similar to Cercospora nicotianae. On potato-dextrose agar the colonies produced by fungi isolated from frogeye spots could not be distinguished from those produced by the fungi isolated from greenspot Altho there was some variation among the isolates the differences were small and occurred in isolates from both greenspots and frogeye spots. The variations were no greater than those commonly oc- curring among isolates in many species of fungi. The colonies were slow growing with aerial gray mycelium in the center and prostrate pink mycelium on the margin. Spores were not produced. The isolates were also cultured on tobacco-leaf-decoction agar prepared by adding 20gms. agar dissolved in 333cc. distilled water to the extract from 200gms. of green tobacco leayes chopped and steamed in 666cc. distilled water.(3) The colonies produced by the two groups of fungi could not be distinguished. On this agar most of the isolates from both frogeye spots and greenspots produced typical Cercospora spores. ‘The spores were similar in size and shape to those produced on natural frogeye leafspots. The conidiophores were longer and straighter than those produced in nature, but there was no difficulty in recognizing them as conidiophores of Cercospora. Inoculation. Potted greenhouse Burley tobacco plants were inoculated with spores produced on agar by spraying a spore suspension in water directly onto leaves. The plants were placed in a moist chamber 3 days, and then returned to the greenhouse. Typical frogeye spots developed within 7 to 10 days. Successful inoculations were made 8 different times. If the plants were incubated in the moist chamber, placed in the greenhouse and then cut and placed in curing chambers on the fifth, sixth, or seventh day after inoculation, typical greenspot developed as the leaves cured. Isolates from either frogeye or greenspot produced frogeye spots on plants kept in the greenhouse, and the same isolates from either frogeye or green- spot produced greenspot if the plants were cut after infection occurred but before visible symptoms appeared. Similar results were obtained in the field. It was found that if leaves were atomized with a spore suspension in the evening at about sunset, infection resulted. If the plants were cut 4, 6, or 8 days after inoculation greenspot developed during curing; if the plants were left in the field, frog- eye developed 12 to 14 days after inoculation. 92 The Kentucky Academy of Science From these observations and tests it is concluded that frogeye on growing ~ and cured Burley tobacco and greenspot on cured tobacco are produced by the same organism, Cercospora nicotianae, under different environmental and host conditions. Bibliography 1 Valleau, W. D. and E. M. Johnson. Tobacco Diseases. Ky. Agr. Exp. Sta. Bull. 362, pg. 37. 1936. 2 Hill, A. V. Cercospora leafspot (frogeye) of tobacco in Queensland, Commonwealth of Australia. Council for Scientific and Industrial Research Bull. 98. 1936. 3 Diachun, S. and W. D. Valleau. Conidial production in culture by Cercospora nicotianae. Phytopath. 31: 97-98. 1941. NUTRITIONAL PROBLEMS IN THE PRODUCTION OF THOROUGHBRED HORSES (abstract) L. J. HorLacHER Department of Animal Husbandry University of Kentucky The purpose of this paper is to summarize some of the investigations - dealing with the nutrition of horses. The leading breeders of Thorough- breds in Kentucky are ever on the alert to utilize scientific findings. Many horses in training develop unsoundnesses. Mitchell states that these dis- abling conditions are merely local manifestations of one general cause. Kintner and Holt found that horses fed a diet in which the calcium- phosphorus ratio was 1:3 were prone to suffer with osteo-malacia. A ratio of 1:1 decreased this difficulty. Greenlee and Beeman by making the same change eliminated impaired gait and stumbling. Buckner, Good and Harms analyzed samples of feed and determined that the feed analyzed was not a contributing factor to leg weakness. The U. S. D. A. has found that the concentration of calcium in the serum of horses does not vary significantly with age, but there is a definite depression of calcium level in the spring. Vitamins are important in horse feeding. Oats and hay are relatively deficient in vitamins A, C, and D. Vitamin A deficiency affects the vision of the horse while its presence helps prevent respiratory troubles. Vitamin D is important in horse feeding because of its value in treating or preventing rickets and other bone disorders. Some practicing veterinarians a Twenty-Seventh and Twenty-Eighth Annual Meetings 93 believe vitamin D deficiency explains the appearance of splints and ring- bones and weaknesses in tendons and ligaments. Most common horse feeds contain sufficient amounts of vitamin E, though there is some evidence that horses may suffer from lack of this vitamin. THE USE OF CORN GLUTEN IN IODINE DEFICIENT DIETS. A PRELIMINARY REPORT (abstract) W. S. Hopexiss Agricultural Experiment Station University of Kentucky During a period of six weeks, diets containing corn gluten and casein as prime sources of protein caused an enlargement of the thyroid gland fzom one to two times those of normal rats. Alternation of beef liver or horse liver in the two basal diets made no significant difference in thyroid size or iodine content. Growth of animals on diets containing corn gluten was slow, reaching approximately 60% of those fed the casein and colony diets. The greatest goitrogenic effect was produced by the diet consisting of corn meal, casein, horse liver and salts. Ash, calcium, and fluorine analyses of the bones of the experimental animals showed no significant variation which could be attributed to iodine deficiency in the rat. 94 The Kentucky Academy of Science NOTES AND NEWS With the publication of this number of the Transactions the first volume of the quarterly series is complete. The Editorial Staff regrets that there has been such a delay in the publication of the present number. Our plans are to have Volume 10 in your hands this year and to have Volume 11, if finances permit, on regular schedule by March, 1944. This journal might well serve as a clearing house of information for Kentucky science and scientists. We therefore invite members of the Academy and all other interested persons to forward news items of general interest, such as promotions, resignations, publications of important books and articles, activities of Kentucky scientists in the armed forces, trips of explorations, etc. Your editors will welcome such information. The Executive Committee of the Kentucky Academy of Science wishes to announce that the special committee for the King Award for 1942 has selected as the winner of the $50 Prize the paper of Martin E. Weeks and Jack Todd entitled, The Semimicro Determination of Magnesium as the Qu‘nolate using the Colorimetric Ferric Chloride Method. Dr. Weeks is an agricultural chemist, Department of Agronomy, in the Experiment Sta- tion, University of Kentucky, and Mr. Todd of the same department is at present on leave for military service. Congratulations to the authors! The 30th annual meeting of the Kentucky Academy of Science will be held Friday and Saturday, April 23, 24, 1943, at the University of Louis- ville. Announcements and requests for papers will be issued in the usual manner. The officers of the Academy for the year 1942-43 are as follows: President ss Seoc00 (2 Noe Sieh ee ee a ee J. T. Skinner Kentucky Agric. Exp. Station NACESPrESICCH ES sores See peer Pa eg ok nae ees L. A. Brown Transylvania College SECKECATY 3 23. oese i tate EN Ue a eg eres Sel Oe ee cea Alfred Brauer University of Kentucky WM reaSUInen:. 10. ces deen” Some OMe oe eee nee eee Wm. J. Moore Eastern State Teachers College . ING oe), Orn (Gromer! NIN Ss Ss ee ee es vee es Austin R. Middleton University of Louisville Councilor tomcyae) union Aca cameron eae ey en ae Anna A. Schnieb Eastern State Weachers College Twenty-Seventh and Twenty-Eighth Annual Meetings VOLUME 9 INDEX Barbour, R. W., A study of the nesting birds of a seventy acre area in Rowan County, Kentucky (abstract) ........... Board of Directors of the Academy, elected at 28th annual meeting . . Carswell, Harry’ E., Some notes cn the diffusible calcium and mag- MESKUMIMETTPAOOdTISeLAN a te ee ye one eee a ck gee Carver, Walter B., The mathematical puzzle as a stimulus to mathe- SINAC C Ae UWVOL Ky ue Toe ee eRe Ve eS ee ie eorrespondence notes. Rare chemicals, ........2)o 5200-2 esd. Davies, P. A., The history, distribution, and value of Adlanthus in IN Gisela ATTEN Garett tos Sx sat eine os ee MBG ose Diachun, Stephen, Relation between frogeye and greenspot of tobacco Stomatal sbenaviorsim sield tobacco) =) ess... 4 es Errington, B. J., Spectrographic identification of minor elements in Hravyz andor eTMOXtUTES os ce ete Ps Sees dca. nike Halle, Louis J., Some notes on the diffusible calcium and magnesium ITAL OOG Sela Meee 2 belt esta Pen tae ag Hazel, Herbert, Resumé of studies on beat tones, combinational tones Ia MISTCLC ATV Seperate es ali ere Ge hes (ON) a oe Rn lire. Charles, The new Iransactions series 2... 02.6.6 ene Hodgkiss, W. S., Spectrographic identification of minor elements in havea! veraimemrxt ress oo $4 See, cp pe dao ae The use of corn gluten in iodine deficient diets AvP te linainanys hepouti (ADSELACE) Init) ere eee Horlacher, L. J., Nutritional problems in the production of Thorough- Bredigiionseses(abStract) nea ike. aes Rt ee ee Jayne, Edgar P., Anatomy of the alimentary canal of Bruchus quad- rimaculatus (Coleoptera) with special reference to the histocytology of the midgut epithelium in the normal (un-fed) and fed conditions (abstract) ............ Johnson, E. M., Relation between frogeye and greenspot of tobacco .. wrmuor Academy, Report of the Councilor ........... 2... 2.222%: Kass, Edward, Enzymic purification of antitoxin .................. Retro iizem Award aO4 Ole ise, Oo Ee Uma sae eat 22 TUSYABL «Seino =A eee GAR AEM cena Ne oy SARE BS en RC perc ea Ee LCMSID GANG MAC TCTSONIA PO eh so, od ee McKenzie, Doris, Hemolytic streptococci from the throats and tonsils of patients with chronic tonsilitis (abstract) ........ ZO} Ese 96 The Kentucky Academy of Science Members elected, 28th annual meeting, 1941 .................... 51 Minutes of the 27th meeting ......2%2..52 0.) 1 Minutes of the 28th meeting’. > =.) 42.) ee 50 Morrison, Sister Charles Mary, Inversion applied to conics .......... 68 Newbury, Edward, The genetics of intelligence .................. 73 Notes: and ‘news (34) he hu ee re 94 O’Bannon, Lester O., On the four-dimensional mechanism of knowing 40 Officers of the Academy 1939-405). 5) 6 es 1 Officers of the Academy 1940-41 22..2 4]. =) eee 49 Officers of the Academy 941-42 2p ee 2. ee §2 Officers of the: Jumor Academy 1941-4222. ee 51 Official Reports: Secretary, Alfred Brauer, 27th annual meeting .... 2 Secretary, Alfred Brauer, 28th annual meeting ....... 50 Treasurer, W. J. Moore, 27th annual meeting ........ 4 Treasurer, W. J. Moore, 28th annual meeting ........ 54 Councilor to the Junior Academy, Anna A. Schnieb, 28th annual aceting 9 ees ee 5 eee 51 Delegate-to S; A: 7A. S:, Morris Scheraco.= ee 51 Payne, V. F.,- Letters to: Robert: Peter) 22.5.1. -. 2 1 88 Research grants, A. A. A. S. fund, 28th annual meeting ............ §2 Resolutions, 28th annual mecting 2 22 52 Ruwe, H. H., The reaction between alkyl sulfates and furoic acid .... 23 Scherago, Morris, Enzymic purification of antitoxin .............. 55 Hemolytic streptococci from the throats and tonsils of patients. with chronic tonsilitis (abstract) .......... 7a Stewart, Olus J., Motion and kinetic theory,- 9.94 = 4) eee 5 Struss, E. F., The reaction between alkyl sulfates and furoic acid... . 23 Sumpter, Ward C., The reaction of nebutylnagnesni bromide with N- cians BP Pere ee Me ae Tie ies - 61 Todd, Jack R., The determination of inorganic phosphorus in corn SEA ws Ld sek ee ee 28 Valleau, W. D., Relation between frogeye and greenspot of tobacco ... 90 Symptoms in “recovered” ringspot tobacco plants and its bearing on acquired immunity (abstract) ........ 14 Vernon, C. C., The reaction between alkyl sulfates and furoic acid ... 23 Warburton, F. W., The role of ‘“‘c” in systems of units ............ 64 Weaver, R. H., Enzymic purification of antitoan - 9... 3a eee 55 Hemolytic streptococci from the throats and tonsils of patients with chronic tonsilitis (abstract) .......... 79 Weeks, Martin E., The advantages of the spectrophotometer for measur- ing the intensity of the blue color in Denigés’ method formphosphorus: and arsemier > ye 81 TRANSACTIONS OF THE KENTUCKY | ACADEMY. OF SCIENCE (Quarterly Series) AFFILIATED WITH THE A.A.A.S. VOLUME 10 (Double Issue) NUMBER 1, JUNE 1942 NUMBER 2, SEPTEMBER 1942 é TWENTY-EIGHTH AND TWENTY-NINTH a ANNUAL MEETINGS e 1941-1942 q : CONTENTS Page “ay wenty-ninth Annual Meeting, Lexington, Kentucky, April 10, and . fi2t942:: Report/of the Secretary. 25 2 ee 1 _ Report Br the Treosurer 1941-420) ee NE Ns Te 4 . The Barrens of Kentucky : Ree ticinteer a. asia Nees Pao ee eee) oes en a" Comparison of Assays of Domestic and Imported Aromatic Plants and Essential Oils Alexandra de Glazoff Paul J. Kolachov ___- RUS eee Vt Ae GM ET Se ey 13 ‘s The Bright Green Bees of the Genera Agapostemon, Augochlora, Augo- { chloropsis and Augochlorella in Kentucky Harvey B. Lovell atid FS) 0, i MR oR 19 © Modification of es cinphacwe in Bruchid (Coleoptera) Embryos by a Treatment with Amino Acids Alfred | Biseterc! 03.0) ea | i Me eee yah AY _ Origin and Development of Prickles of Aralia Spinosa ee eens oR meh tees ke NOR eb gee 29 TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE EDITORIAL STAFF Lawrence Baker .......... Berea College ..... Psychology & Philosophy ; BW COOK A ane Centre ‘College. 0.1 VLA es Bacteriology — at Bewovell ys) Meuct was University of Louisville ........... Biology { ALG. McFarlane University of Kentucky ....... .... Geology Ward: ‘CG. Sumpter, 3235. Western State Teachers College ... Chemistry jarvis’ Dodd eek ale well University of Kentucky ..-..:..... Physics — Jolin (Kariper: uae University of Kentucky .. Managing Editor Manuscripts. The Transactions must be limited to the proceedings of the annual meetings of the Kentucky Academy of Science and to original manuscripts pertaining to science. Manuscripts are subject to the approval of the Editorial Staff and may be submitted to Editor of the subject cov- ered or to the Managing Editor. Extra-Cost Features. ‘The extra cost of special features such as cuts, graphs, tables, etc., above the text-run price per page must be borne by the contributor. The Editorial Staff will advise contributors concerning the extra cost of features upon receipt of manuscript. Illustrations to be included in an article should accompany the manuscript if possible, or, if sent in separate package should be properly labeled as to the article i in which they are to occur. Proor. Galley proof will be sent for approval of contributors. The proof should be returned promptly to the Berea College Press, Berea, Kentucky. REPRINTS. Reprints are furnished at publisher’s prices by negotiating directly with Berea College Press. Price quotations on reprints are sub- mitted with the proof. Orders for reprints should accompany the proof to the Berea College Press, Berea, Kentucky. SUBSCRIPTION Rates. The Transactions is sent without additional ex- pense to all members of the Kentucky Academy of Science who are not in arrears for annual dues. The annual subscription rate for non-members is $2.00 in the United States and Canada, $2.50 in foreign countries; single number 75 cents. One volume of four numbers appears each Academy fiscal year. BusINEss CORRESPONDENCE. —— and correspondence concern- ing subscriptions, extra costs, ren matters except reprints should be addressed to Fis ee f£ Kentucky, Lexington, Kentucky. 4 s % SS 4 . cro) ¥ seh 4 ae SNA wust Twenty-Eighth and Twenty-Ninth Annual Meetings 1 TWENTY-NINTH ANNUAL MEETING held at University of Kentucky, Lexington, Kentucky April 10 and 11, 1942 First Business Session Called to order by President Pennebaker at 1:45 P.M. After the opening remarks by Dr. Pennebaker the following reports were heard: Secretary’s report, accepted and ordered filed. Treasurer’s report, received and accepted. Report of Anna A. Schnieb, Councilor of the Junior Academy. Report of Morris Scherago, delegate to Southern Association for the Ad- vancement of Science and Industry. This Association was formerly known as the Southern Association for the Advancement of Science. A motion was made, seconded and passed that an invitation be extended to the S.A.S.J. to meet with the Kentucky Academy of Science at a future meeting and that the matter of attending to the arrangements for such meeting be ‘referred to the Executive Committee. A report of the Auditing Committee was read as follows: “Richmond, Ky., April 9, 1942. To the Kentucky Academy of Science: We have found the treasurer’s books to be in perfect order and recommend the acceptance of this report. It is a pleasure to audit accounts kept so neatly and accurately. Signed, A. D. Hummer, Chmn. L. G. KENNAMER. Other reports called for were those of the Membership Committee, and the Committee on Kentucky Wild-Life and Natural History Conference. The chairmen of these committees respectively requested that their reports be deferred to the second business session. The requests were granted. Second Business Session 8:00 A.M. April 11 Call to order by President Pennebaker. A report by Dr. A. R. Middleton, Representative on the Council of the A.A.A.S., was heard. The report provoked an active discussion relative to the distribution of the research grants and on the disposition of the grants made by the grantees. On completing the report, Dr. Middleton made a motion that the recipients of grants be asked to report at the Annual Meetings of the Academy as to the state of their research for which the aid was granted, until the work is terminated by its publication. The motion carried. 2 The Kentucky Academy of Science The Secretary commented that the terms of this motion had been met in the past, ever since the grant was instituted. Dr. B. B. McInteer now gave the report for the Committee on Kentucky Wild-Life and Natural History Conference. At the conclusion of the re- port he made the motion that the Kentucky Academy of Science solicit the affiliation of this group, and that the Executive Committee take the matter into consideration. Motion was carried. Dr. Keller, Chairman of the Membership Committee then prea the following for membership, all of which were elected: Astle, Melvin J., Dept. Chemistry, U. of Ky. Baker, Lawrence M., Dept. Psychology, Berea College. Dekker, Albert O., Dept. Chemistry, Berea College. Hart, Julian M., Dept. Philosophy, Berea College. Jayne, Edgar P., Flat Gap, Ky. Lincicome, Dr. David Richard, Dept. Zoology, U. of Ky. Shannon, E. L., Dept. Chemistry, U. of Ky. Wender, Simon H., Dept. Chemistry, U. of Ky. “Smith, William Landon, Dept. Zoology, U. of Ky. Dr. Leggett, reporting for the Resolutions Committee, proposed that the following resolutions be adopted: 1. That in consideration of the excellent work of the Junior Academy of Science under the guidance of Dr. Anna A. Schnieb, the Kentucky Academy of Science urge its members to hold themselves in readiness to give whatever assistance they can to the Junior Academy. That the Kentucky Academy of Science express its appreciation for the successful efforts of Dr. Schnieb in building up the Ky. Junior Academy of Science and commend the young people of the state for the excellent work that they are doing. 2. That the membership of the Kentucky Academy of Science notes with pride the fact that so many of our members are in the service of our country during its present emergency, and urges the proper officials to keep a record of the members and the services they are giving; and, that the Academy do everything practicable to bring victory for our armed forces at the earliest possible time. 3. That the Kentucky Academy of Science, through its President and Secretary, express its thanks to the administration and faculty of the Uni- versity of Kentucky for the use of buildings, and for their hospitality on this the 29th Annual Meeting of the Academy. Signed, LuciEN Beckner; J. H. Capps; J. L. Leccrrr. The resolutions were adopted. . | W. R. Allen, reporting for the Nominating Committee, presented the committee’s nominations for officers. President Pennebaker reminded the assembly that nominations from the floor were in order, and that any mem- ber of the Academy had the privilege of making a nomination for any Twenty-Eighth and Twenty-Ninth Annual Meetings 3 ofice. No other nominations were made. The Academy then cast its vote for the entire slate as presented. This is, President, J. T. Skinner, Western Ky. State Teachers College Vice-Pres., L. A. Brown, Transylvania College Secretary, Alfred Brauer, Univ. of Kentucky Treasurer, Wm. J. Moore, Eastern Ky. State Teachers College A.A.A.S. Council Representative, Austin R. Middleton, U. of Louisville Directors to serve till 1936, J. H. Capps, Berea, and H. P. Sturdivant, Barbourville. The Academy then adjourned to the various divisional meetings. Signed, ALFRED BrAvER, Secretary. MINUTES OF THE EXECUTIVE COMMITTEE 1941-1942 First Meeting This was held just after adjournment of the Twenty-Eighth Annual meeting, April 26th, 1941, at Richmond, Ky. All members were present. No official business was transacted. It was agreed that the second meet- ing would be held in Lexington, sometime during the summer, upon call by the President. Second Meeting Held at Lexington, Aug. 14, 1941. Those present were: Pennebaker, Hire, Skinner, Moore, and Brauer. The matter of placing the King Award was discussed. The plan now followed is that the divisional chairmen with the aid of assistants which they may choose, select the paper from their respective divisions which they consider most worthy. The divisional chairmen then constitute the com- mittee which determines the final award, from the papers selected in the several divisions. The members of the committee respectively rate these papers in regard to a number of established points, after which the chair- man of the general committee submits the selection to the Executive Committee. Dr. Hire then outlined his plans for the publication of the Transactions. The place of the next annual meeting was left undecided, but was tenta- tively but unofficially set for Bowling Green. The following were elected to membership: Carl Wade, Western Ky. S. T. C.,, Bowling Green Bruce Rawlings, Dept. Botany, U. of Ky. Transactions by Correspondence — King Award In October 1941, the Executive Committee approved the selection of the Committee on the King Award. The selection was announced in the press 4 he The Kentucky Academy of Science and in Science. Mr. and Mrs. Fain W. King were likewise informed of the selection. The title of the paper and the authors are, “Enzymic Purification of Antitoxin”’—- Edward Kass, M. Scherago and R. H. Weaver. Place of 1942 Meeting Established In January 1942, the Executive Committee set the place of the Twenty- Ninth Annual Meeting at the University of Kentucky, Lexington, for April 10th and 11th. This was done by ballot SULTS to the members of the Executive Committee, by mail. Third Meeting April 10, 1942 There were no requests for research aid. It was moved that the Secretary request the Secretary of the A.A.A.S. to carry this grant over to the fol- lowing year. (The amount is $44.00). Motion carried. Dr. Schnieb requested an amount of $30.00 for the Junior Academy for the year. The motion was made that this amount be allowed. Carried. Dr. Schnieb made the motion that the President of the Academy annual- ly appoint District Supervisors, whose duty it shall be to look after the interests of the Junior Science Clubs in their respective districts. The motion passed. President Pennebaker then appointed the following supervisors: First Dist —W. B. Mozier, Murray, Ky. and Jas. Major, Paducah. East. Ky. Dist.—Lucille Catlett, Morehead. Fifth Dist.—Mrs. Rethwisch, Anchorage. Northern Dist.—Austin Durham, Brooksville. Mid-Cumberland Dist.—H. E. Allen, Somerset. Central Dist.—Miss Anne Foley, Richmond. Upper Cumberland Dist.—Robert Watson, Lynch, Ky. Signed, ALFRED BRAUER, Secretary. REPORT OF THE TREASURER OF THE KENTUCKY ACADEMY OF SCIENCE FOR THE YEAR 1941-42 (As of April 10, 1942) INCOME Balance in Bank at Beginning of Academy) Year, April) 26,1941. $664.50 From Dues: For the Year 1939-40 For the Year 1940-41 For the ‘Year 941-4232. a oo 294.00 360.00 From the Junior Academy—Fees ....... 106.35 UAUNARS Gra Ni eee oe MEN NN Slee lee ty ailege AU 75.00 Twenty-Eighth and Twenty-Ninth Annual Meetings Reprints: Bima Alominn. > iy ieee Ssh) eka dona 8.75 eles GOVE ls 8 oc ceeh oh i inlet Siu a ee 4.00 2765) Dividends: Lexington Federal Savings and Loan PNSSOCIALIONN ee hi Meta Cn ue yates 29°99 ene CaP ANW iat tame coh ne en lo GU Hs a 50.00 DISBURSEMENTS Mr. Fisher, Speaker at Annual Meeting, TE earl Bei ap a ei aka eh etapa oem eg $ 2.00 The Kentucky Kernel: Printing Programs for Annual Meeting $ 16.50 Printing of vlransactions .../.: 0.0... 168.00 eprints and Envelopes ..........:.. 16.50 Printing Stamped Envelopes ......... 18.02 219.02 B. B. McInteer—A.A.A.S. Grant ....... 25.00 Ward C. Sumpter—A.A.A.S. Grant ..... 25.00 miatherine Carr—A.A.A.S. Grant ....... 25.00 Alfred Brauer: SPAMAP Ss MCLOI i game eee Mla 13.50 Mimeograph Material and Postage ..... 4.50 18.00 Treasurer of Berea College: Printing Transactions and Incidentals.. 79.83 monte: (Pransactions 7204.94: - NODES 189.36 For the Kentucky Jr. Academy of Science: Berea College Treasurer for Printing 3 issues of Jr. Bulletin-: ..).......%. 098 Thomas Cole Phelps, Treasurer of. Junior Academy—Part Payment on Five Cent TEGREM: py GS AEN ke OER cy SEE rea ras 30.00 Anna A. Schnieb, Councilor—Balance on junior Academy, Heesis 2 2. 5.45 Anna A. Schnieb, Councilor—Balance on Contribution from Academy ...... S537, 136.35 W. J. Moore—800 Postal Cards ........ 8.00 M. Scherago, R. H. Weaver, and Edward @ass——Kine Award fs 26.30. ve 50.00 RAM EIIC EMT UL Eee Sid ear Ns etal epi, $1298.59 697.73 600.86 Respectfully Submitted W. J. Moore, Treasurer 6 The Kentucky Academy of Science FINANCIAL STATEMENT OF THE KENTUCKY ACADEMY OF SCIENCE IN CONNECTION WITH THE JUNIOR ACADEMY OF SCIENCE April 10, 1942 INCOME From Junior Academy Membership Fees .......... $106.35 Contribution from Senior Academy .............. 30.00 $136.35 DISBURSEMENTS Berea College Treasurer — Printing three issues of the} Junior Academy Buller. ie $95.53 Thomas Cole Phelps, Treasurer of Junior Academy— Part payment on five cent) tees. 4) c)0) ee 30.00 Anna A. Schnieb, Councilor — Balance on Junior Aicademiy thees (unc Wi \ay Min Dae an ane tO net inn i ae 5.45 Anna A. Schnieb, Councilor — Balance on Contri- bution\“from: Alcademayy) 00s | aa Gn ee arena 57 136.35 Balance yy ii ieee iil een aa ee We NLR ni yas MCSE I eae 0.00 Respectfully Submitted W. J. Moore, Treasurer The Kentucky Academy of Science Note: It can be seen from this statement that the actual cost to the Senior Academy, in its sponsorship of the Junior Academy for the present year, is $30.00. Twenty-Eighth and Twenty-Ninth Annual Meetings 7 THE BARRENS OF KENTUCKY B. B. McINTEER Department of Botany University of Kentucky The name given to a certain part of Kentucky by the early settlers served as a description, according to their ideas. They found in some parts of this state, open grassland with only occasional trees which were generally too small to be used very well for building material, fences, and fuel. They gave the name “Barrens” to this region because they thought the absence of trees signified that the soil was non-productive or barren. Concerning this, Shaler (15) wrote the following: At first the white immigrants conceived a strong prejudice to this untimbered ground, deeming the absence of trees as evidence of the poverty of the soil. The pioneers had always used trees as criteria of the productivity of the soil—associating certain species with definite soil quality—and when they reached a territory destitute of trees they obviously thought that the soil was of little value and, in fact, for this reason, as well as others, they avoided settling in the grassland. Dicken (4) called attention to the fact that the first important settle- ments of this region were in the wooded territory just outside the Barrens or on streams (near which timber grew) that traversed the Barrens. These towns in due time became county seats. They are Brandenburg, Elizabeth- town, Munfordville, Bowling Green, Hodgenville, Glasgow, and Russellville. The fantastic tales told of this region gave the idea that the Barrens were so dry and hot that it was beyond the ability of man to endure them. Michaux (11) who had heard some of these stories was agreeably surprised when he entered the Barrens, for he wrote: I conceived I should have had to cross over a naked space, sown here and there with a few plants. I was confirmed in my opinion by that which some of the country people had given me of these immense meadows before I reached them. They told me that in this season I should perish with heat and thirst, and that I should not find the least shade the whole of the way. . . . Instead of finding a country as it had been depicted to me, I was agreeably surprised to see a beautiful meadow, where the grass was from two to three feet high. According to the ideas of Sauer (14), the English-speaking people had no word for land covered with native grass, for they had never seen such a condition. He brought out the fact that after grasslands were settled in the states north and west of Kentucky the term “Prairie” was adopted from the French language. Owen (13) seemed to think that the appearance of the soil as well as the nature of the vegetation served as a basis for the name given to this region, for he wrote: This limestone group (Barren limestone) produces, for the most part, an excellent soil. . . .In the early settlement of Kentucky the belt of country over which it extended 8 The Kentucky Academy of Science was shunned, and stamped with the appellation of “Barrens”; this arose, in part, from the numerous cherty masses which locally encumbered the ground. Weir (18) seemed to think that the name was due to the absence of timber, for he wrote: It is a well known fact, that a greater portion of this part of Kentucky, and that, too, in the last half century, was nothing more or less than a real, genuine, bona fide prairie. The Barrens are generally referred to as grassland, but, in reality, grasses made up only a part of the flora, for there were composites, sedges, woody vines, shrubs, and scattered trees of a scrubby growth. According to oral tradition, one could often ride for miles and not find a riding switch. Gar- man (7) gives a list of a few plants found within the Barrens, such as: grasses, rushes, golden rods, asters, shooting stars, dwarf willows, black jack, tall bluestem, Indian grass, prairie clover, button snakeroot, blazing stars, and compass plant. Gorin (8) wrote: “The Barrens in the spring, summer and fall were covered with millions of the most beautiful and variegated flowers.” Haycraft (9) mentioned the fact that in some sections of Hardin County the Indians, when attacking the whites, hid in the grass and cane, for there were no trees. He further wrote that the horses and cattle had good pasture, for they browsed on the cane brakes which showed a luxuriant growth on all rich land, especially near the water courses. However, he explained that along the creek on which Elizabethtown is built there was a good growth of timber. Hussey (10) made some very striking statements concerning his. im- pressions relative to the northern part of Barren County. He wrote: My observations in Barren County would lead me to the conclusion that the traditions which are current as coming from the settlers are true; that is to say, that when the whites first came to these parts, it was, indeed, a barren region, destitute at least of trees. On the more level parts of the county the trees are yet small in size and few in species. For the most part, the main region of the Barrens (Big Barrens) cor- responded with the area of Cavernous limestone, forming a rather narrow band extending from the Ohio River just west of Louisville southward to the Tennessee State line and westward to the Cumberland River. All this land was not covered with grass, however, for in certain sections, especially along streams, there was a good growth of forest vegetation. Furthermore, some territory other than the Cavernous limestone was covered with grass, but the extent was not very great, and it was always adjacent to this par- ticular geologic region. Since each of the early writers saw only a portion of the Barrens, it is necessary to summarize the references of a great number of them in order to get a clear picture of the region actually covered with prairie vegetation. Michaux (11), when writing of his travels in 1802, explained that he en- tered the Barrens on the northeast, near the Little Barren River, and left the region when he crossed the Tennessee State line at which place he en- tered a wooded territory. It is thought that he crossed from Allen County, Kentucky, into Sumner County, Tennessee, about forty miles northeast of Twenty-Eighth and Twenty-Ninth Annual Meetings 9 Nashville. Regarding the size of the Barrens, he made the following state- ment: “The Barrens or Kentucky Meadows, comprise an extent from sixty to seventy miles in length, by sixty miles in breadth.” This was the main part of the Barrens extending from the Green and Little Barren rivers to the Big Barren River but ne wali (he gave is too great, for perhaps the widest part was not more than twenty-five miles. According to Shaler (15), the Barrens were in the region of the present L.&N Railroad which for most of the way, while in Kentucky, is within the limits of the Cavernous limestone. Cotterill (3) describing the condi- tions met by the early settlers, wrote: But there was one section of Kentucky whereon no forests grew. An area of six thou- sand square miles lying immediately south of the Falls of the Ohio was destitute of trees, though over-abounding in grass. The American Universal Geography by Morse (12) has the following comment: In Nelson County, northwest of Rolling fork, a branch of Salt river is a tract about 40 miles square, mostly barren, interspersed with plains and strips of good land. Perhaps this corresponds to the northern half of Hardin County and the southeastern two-thirds of Meade County. Brown (2) explained that near Russellville the strip of grassland was fifteen miles wide and that it extended from east to west for a distance of ninety miles. Gorin (8) mentioned the fact that that part of Barren County north of Glasgow was covered with grass while the southern part was covered with timber. Dicken (5) described the location of the Barrens as follows: Southwestward from Louisville, flanking the Western Coal Basin in a broad crescent lies the greater part of the Kentucky karst, a streamless region known since the early days of settlement as “Big Barrens.” The early writers show the extent of the Barrens and the conditions that were prevalent when the whites first came to this part of the country, and while they give their own ideas, they do not suggest the real cause of such conditions in the vegetation, for that which was responsible for this sit- uation dates much farther back than the time of the first pioneers to Ken- tucky. Concerning the origin of the Barrens, one should take into con- sideration the natural causes, which may be climate or soil or both, as well as the cultural practices of the men who were present. Many early writers thought that frequent fires were the cause of the peculiar vegetation. Michaux (11) said that the whites were following the customs of the Indians by burning the grassland in early spring of nearly every year, first to attract the game into the open, and later to make better pasturage for the cattle. He stated: Every year, in the course of the months of March or April, the inhabitants set fire to the grass, which at that time is dried up, and through its extreme length, would conceal fiom the cattle a fortnight or three weeks longer the new grass, which then begins to spring up.. Bourne (1) argued that while the prairies in the states farther north 10 The Kentucky Academy of Science were caused by water conditions, the grasslands of Kentucky are different and their origin is different. He said: The soil is not of recent alluvion like the prairies; ...I think it must be evident to everyone who will view the Barrens attentively, that their present appearance was caused by fires, which have consumed the trees and the acorns from which they grow: because many of the trees that are standing are partially burnt, and almost everyone that is lying down has been burnt more or less. Shaler (15) apparently thought that the customs of the Indians were the main cause of the grassland. He thought that the region had been covered with trees but that the Indians by frequent burnings destroyed these woody plants and permitted the grass to grow, for he wrote: This destrtuction of the timber was brought about by the custom, common to Western Indians, of burning the grass of cpen ground and undergrowth of the woods, in order to give a more vigorous pasturage to the buffalo and other large game. Sauer (14) seemed to think that the underlying rock had something to do with the presence of the grassland. He said: The partial correlation between grassland and cavernous limestones has suggested a causal connection between the nature of the soil and the vegetation... On the other hand, Transeau (17) maintained that the factors of climate, especially humidity, were a contributing cause. His accompanying map shows that “the average relative humidity—July” indicates that the West- ern Coal Field, Chester, and Cavernous limestone regions are in one zone while the rest of the state is in another, with a higher humidity. Since the region of grassland so closely coincides with a narrow strip of land underlain by cavernous limestone, it seems that the nature of the underlying rock was the main contributing cause. Still, Transeau showed that the relative humidity is lower in that region than in other parts of the state, and he brought out the fact that this low humidity is favorable to the development of grassland. The gradual change in this climate is not sufficiently pronounced to cause such an abrupt change in the vegetation at the border lines between the Cavernous limestone and the two adjacent regions, according to the ideas of some. There was a severe drought in this part of the country about 3,000 years ago and that, perhaps, was the first step in bringing about the conditions that the pioneers found in the Barrens. It seems that trees on land under- lain by a thick, compact, limestone layer, from a few inches to several feet below the surface, cannot endure a drought as well as can the same type of plants on sandy soil. There may be many reasons for this difference. Dicken (6) brought out the fact that within the Cavernous limestone region the surface water courses and true valleys are almost absent, since the run-off quickly flows or seeps downward in the hollows and drains away thru the extensive underground circulation. It is very noticeable that when a region has underground drainage the soil becomes drier than that in a similar region with surface drainage only. The texture of the soil in the various regions, especially under abnormal conditions, may be the determining factor in the: type of vegetation that will survive. Shantz (16) revealed some very interesting facts: Twenty-Eighth and Twenty-Ninth Annual Meetings 11 Sand is also an equalizer when the physiological conditions of soil moisture that affect the plants are considered. In a region of heavy rainfall, drainage in sand is so rapid that the plant is left in a moist soil with plenty of aeration. In regions of little rainfall the scant moisture supply passes rapidly into the soil which, because of its low water-holding capacity, allows it to penetrate much deeper than in heavy soil, and this moisture is pro- tected from evaporation loss by the sand layer above. The soil moisture under both con- ditions is almost identical. In extremely dry conditions water, by means of capillarity, rises through the sandstone but can scarcely, if at all, rise through the limestone. Fur- thermore, the roots of plants can penetrate the sandy soil to a much greater depth than can be done in the clay that is associated with this particular limestone. Dicken (5) explained that limestone is almost impermeable and that water cannot sink farther than the upper part of the limestone layer unless it goes into open joints. If these rocks are practically impermeable to the water above them, it is true that they are likewise impermeable to the water below, if present. Furthermore, due to underground streams, the limestone region is unable to retain the water as well as can the regions having surface drainage. The foregoing paragraphs merely explain how the treeless condition was, probably, brought about, but do not explain directly the presence of grass. The grasses and other herbs undoubtedly invaded that region as soon as conditions became favorable for their growth and reproduction. There were two sets of ideas as to the reasons for the land’s remaining in grass. Some claim that the climate, especially the extremes that were ex- petienced, was favorable to the growth of grass—not on all land but on that which had a tendency to be thirsty—and that this alone is sufficient to explain the situation. Others, however, advance the explanation that frequent fires were the cause, wholly or in part, for fires which might entirely destroy other plants could scarcely hurt those herbaceous perennials having underground stems from which shoots could readily grow. Any careful observer who is familiar with the conditions can readily see why fires were so much more effective in the Cavernous limestone region than in other sections of the state. With few exceptions, there are no surface streams of water in this region; they are mostly underground. When « fire is started in this region it does not meet with natural bar- riers, such as streams of water. The topography as well as the presence of surface streams in other sections of the state would influence the extent of range covered by a fire. A fire moves very slowly down hill, and, under such conditions even a very small stream of water would serve as a barrier. The whole problem is a complex one . The writer is of the opinion that the presence of the Barrens is due to the combined influence of the peculiar soil, climatic conditions, and fires. Literature Cited 1. Bourne, A. 1820. On the Prairies and Barrens of the West...Am. Jour. Sci. & Arts (Silliman’s Journal) 2:30-34. 10. Wes TRGE The Kentucky Academy of Science 12 2. Brown, Samuel R. 1817 Western Gazetteer. H. C. Southwick, Auburn, NGS 3. Cotterill, R. S. 1917. History of Pioneer Kentucky. Johnson & Hardin, Cincinnati, Ohio. 4. Dicken, S. N. 1935. The Kentucky Barrens. Bull. Geogr. Soc. Phila, 43: 42-51, illus. ; 5, 1935. Kentucky Karst Landscapes. Jour. Geol. 43: 708- 728. 6. ———————— 1938. Soil Erosion in the Karst Lands of Kentucky. U. S. Dept. Agr., Soil Conserv. Cir. 490. illus. 7. Garman, Harrison. 1925. The Vegetation of the Barrens... Trans. Ky. Acad. Sci. 2: 107-111 . Gorin, Franklin. Times of Long Ago. Barren County, Kentucky. (Re- print of series published in Glasgow Times. 1876) Published by L. H. Gorin, Louisville, Kentucky. 1929. . Haycraft, Samuel. A History of Elizabethtown, Kentucky, and its Surroundings. (Written in 1869). Published by the Woman’s Club of Elizabethtown, Kentucky. 1921. Hussey, John. 1876. Report of the Botany of Barren and Edmonson Counties... Ky. Geol. Sur. 1: 27-58. . Michaux, F. A. 1805. Travels to the Westward of the Alleghany Mountains in the States of Ohio, Kentucky and Tennessee... Un- dertaken in the year 1802... Transl. from original French by B. Lambert. London. Printed for Richard Phillips. . Morse, Jedidiah. 1793. The American Universal Geography. Isaiah Thomas and Ebenezer T. Andrews. Boston, Massachusetts. . Owen, David Dale. 1856. General Report of Geological Survey. Made during the years 1854 and 1855. Ky. Geol. Sur. 1: 15-248, illus. . Sauer, Carl Ortwin. 1927. (Assisted by John B. Leighly, Kenneth Mc- Murry and Clarence W. Newman) Geography of the Pennyroyal... Ky. Geol. Sur. 25: 303 pp., illus. . Shaler, N. S. 1885. Kentucky; a Pioneer Commonwealth. Houghton, Mifflin and Company. New York. . Shantz, H. L. 1938. Plants as Soil Indicators. Yearbook of Agriculture. U. S. Dept. of Agr. 835-860. Transeau, E. N. 1935. Prairie Peninsula. Ecology 16: 423-437. Weir, James. 1850. Lonz Powers or The Regulators. Lippincott, Grambo & Co. Philadelphia, Pennsylvania. Twenty-Eighth and Twenty-Ninth Annual Meetings 13 COMPARISON OF ASSAYS OF DOMESTIC AND IMPORTED AROMATIC PLANTS AND ESSENTIAL OILS ALEXANDRA DE GLAZOFF and Pau J. KoLacHov Joseph E. Seagram & Sons, Inc. Louisville, Kentucky The war in Europe has made necessary a search for new sources of supply of aromatic plants for essential oils. Aromatic plants have an extensive use in various branches of industry. From the records of foreign commerce and navigation of the United States for the year 1938, we find that over 200,000,000 pounds of spices and drugs costing $19,000,000 were imported. The gin industry alone uses 500,000 pounds of those aromatic plants per year. This large quantity of aromatic raw material has been imported from all over the world. Coriander seed was imported mainly from Hungary, Russia and Africa (Morocco), Angelica root from Germany (Saxony), Lavender flowers from England and France, and Juniper berries from Italy (Toscana). After the usual foreign sources became scarce, the search in this country followed two courses. The first was a campaign for introducing the cultiva- tion of the foreign species in this country while the second was the intense study of all wild domestic sources of species in demand, with hopes of using them until the results of cultivation became available. As results of the work in the first direction, two botanical species formerly imported in great quantities from abroad have been cultivated in this country, mainly here in Kentucky. These two species are: Coriander and, to a smaller extent, Angelica. As a result of encouraging analyses of various _ samples of the 1940 crop obtained in our laboratories a second year cultiva- tion of Coriander began in this state on a larger scale this year (1941). On the other hand, the study of existing wild species or those species culti- vated on a small scale furnished quite interesting results.) Among those could be mentioned Lavender and Juniper. While considering the results of analyses of all four mentioned plants, we shall give a brief description of each. I. CORIANDER Coriandrum sativum L. is an annual Umbelliferae. The commercially important part of Coriander is a grayish yellow fruit commonly called seed. When collected fully ripe, it has a sweetish taste and a pleasant aroma due to the presence of an essential oil. The unripe seed has a very un- pleasant odor and taste. The main chemical constituent of Coriander oil, which represents between 0.2 and 1.2% of the seed, is linalool present in amount of 60-80%, and its acetic esters. The other known constituents are d-l pinene, n-cymol, alpha gamma and terpinene, geraniol acetic esters, 1-borneol acetic esters, and depentene or limonene. 14 The Kentucky Academy of Science Coriander is widely used. It finds applications in food industries as a spice or condiment, in flavoring tobacco, for pharmaceutical purposes, in making liquors, and particularly in gin manufacture. Oil is used in perfume and soap industries; Coriander oil is extremely valuable because five various aromas can be developed from it. Linalool gives the odor of lily of the valley, linalyl acetate gives the odor of bergamot, citral (which comes from limonene) gives the odor of lemon, ionone methyl ‘ionon gives violet odor, nerol-geraniol gives the odor of rose. Coriander, as well as the other botanicals, has been analyzed by our usual method which consists in determinations of moisture by desiccator and oven, acidity, refractive index, contamination, quality of single distillate, and amount of oil by two means. The first one is a Seagram modification of Clevenger’s’ method, consisting of steam distillation combined with cohobation; the second is a modified Johnson’s ether extraction method. The analyses of Coriander showed a domestic crop of remarkable qual- ity. Three types of Coriander have been planted in Kentucky, Hungarian, Russian, and Moroccan; in each case the analyses have been compared with the original imported samples. The results are shown in Table I. TABLE I Analyses of Coriander Seed (Coriandrum sativum L., Umbelliferae) IExpaeNo: seo ese 491-3 491-5 491-2 508-1 491-1 516-2 AB eke e Bk ee 1939 1940 1939 1940 1940 1940 Country =s-—— ese Russia USA(Ky.) Hungary USA(Ky.) Morocco USA (Ky.) grown grown grown from from from Russian Hungarian Moroccan Moisture by desiccator —_______ 2.748 % 2.967 % 2.788 % 3.536 % 2.709% 3.061% Moisture by : oven) 222-2 _---- 6.388% 6.900% 6.510% 5.990% 6.042 % 4.980 % Total Moisture ___ 9.186% 9.861% 9.298% 9.525.% 8.751% 8.041% Oil by Seagram’s Method —_-_-___-__ -98% 1.02% 89% 57% -25% .35 % Oil by Clevenger ; Method -_---___--- 1.00% 1.00% -90% -60% -25% 40% Acidity as mg. KOHi/smy anes ese 1.38 2.14 1.75 1.33 Pap 2.52 Refractive index nD 20°/20°C ____ 1.46387 1.4630 1.4640 1.4643 1.4626 1.4631 Odor (=) 22a Similar Similar Similar From analytical results shown in this table, it appears that: 1. Russian grown Coriander has slightly more oil than the Hungarian. Both Hunearan and Russian grown seeds have 3/2 to 4 times more oil than the Moroccan seeds. 2. Russian and Moroccan Coriander planted in this eres have a higher oil content as well as higher acidity than the original seeds used for planting. 3. The refractive index is very closely related. 4. A close similarity in flavor has been found. Twenty-Eighth and Twenty-Ninth Annual Meetings 15 Il. ANGELICA—Angelica archangelica L., Umbelliferae It is the second important plant beginning to be cultivated in this country. It belongs to the same family as Coriander; however, Angelica is a biennial. Before the war Angelica was imported mainly from Ger- many, (Saxony), Hungary, and Belgium; while second grade material was imported from Japan. The commercially important parts are dried root containing 0.3-1%, and seeds containing 0.7-1.5% of essential oil, although the essential oil is distributed almost throughout the whole plant. The most important chemical constituent of Angelica oil is d-phellandrene. The other constituents found were iso-valeric acid, valeric acid and esters, - oxymyristic acid and esters, pinene, angelicine, angelic acid, and oxypenta- decylic acid. The lactone of oxypenta-decylic acid seems to account for the musky odor of the root oil. The seed oil does not possess this compound. It has mainly d-phellandrene and esters of valeric and oxymyristic acids. The Angelica also has various applications. Small roots, young stems and leaves are used in candy manufacture. Whole root is used in flavoring blending tobacco and in the Vermouth wine and gin industry. The seeds are used in pastries and beverage industries. The essential oil of the root is used in perfumes, bitters, and cordials. Our laboratories had the opportunity to analyze several samples of do- mestic Angelica, and whereas the wild samples do not yet compare favorably with our standard, the cultivated sample, from North Carolina, showed very promising results. At present more Angelica is under experimental cultivation in Kentucky. Table II shows how these various samples compare. TABLE If Analyses of Angelica Root (Angelica archangelica L., Umbelliferae) IZM OINO. yah oe LS 100-1 508-4 462-2 515-5 522-3 ce Se ee eee 1940 1941 1940 1941 1941 @ountry;) 222-22 =. | Hungary Japan Germany, USA, Ky. USA, Saxony (wild) N. Carolina (cultivated) Moisture by Gesiceator 22 ——_ =~ 2.330% "2.224% 344% -3851% 3.770% Moisture by DG See eee 8.200% 4.998% 5.866 % 5.801% 5.448% Total Moisture _____ 10.530% 7.222% 6.210% 6.192% 9.218% Oil by Seagram’s Menhod) 22222-2222) 59% -35 % 80% -711% -80% Oil by Clevenger Wethod 2). 222 Sue -35% A4% CDI BV wa eae -85% Acidity as mg. KOH /em > 222. - = 9.5 7.56 7.42 6.51 4.34 Refractive index G20 / 20°C) shes 1.4842 1.4838 UO Oi ue Lip tgs ee 1.5348 Similar to Otte fo ae ne Different Saxonian Slightly similar to Japanese 16 The Kentucky Academy of Science 1. The yield of oil in domestically cultivated species is almost the same as in the best imported Saxonian type. 2. The domestically cultivated species has more oil than the wild. 3. The acidity and refractive index of the two (best imported and best domestic) are quite different. 4. The flavor of the above mentioned samples was similar. 5. The flavor of wild species was very unpleasant. Ill. LAVANDULA VERA D. C. Our laboratories spend considerable time continuously trying to intro- duce new ingredients in our products. One such ingredient is Lavender flowers. It has just been found that there was a possible chance of intro- ducing this ingredient when the foreign supplies were cut off and we were forced to stop our experimentation with Lavender. Recently, we received a sample of domestic cultivated Lavender from Texas. The analyses showed possibilities for using and cultivating this plant on a larger scale. Recently, it was learned that a larger Lavender scale experimental cultivation of this plant is being conducted in the State of Washington near Seattle. Lavender is a small perennial Labiatae. The commercially important parts of Lavender are small bluish flowers containing from 1 to 4% of essential oil. The main chemical constituent is I-linalyl acetate (30-45%); other constituents are free linalool, geraniol, pinene, and cineol. The oil of Lavender flowers is used in perfume, soap industries and med- icine. It is an excellent melliferous insecticide and finds uses in the cer- amic and lacquer industries. The by-product of dried Lavender flowers is used for cattle feed. TABLE III Analyses of Lavender Flowers (Lavandula vera D. C., Labiatae) BEY SSP) SoG cs ow EN ok 520-4 508-5 UA tei ee nde ee ENE MSDE ea nS ae RD as 1940 1940 COUT E ry ee EY ay BL TS England U.S.A. Mitcham Scottsville, Texas Moisture by desiccator ~~-_____._-____ 1.190% 7.333 % Moisture by oven ______--____________ 5.583% 9.250% Total Moisture: 2b 2 wee eee ee ee 6.773 % 16.583% Oil by Seagram’s Method —__._______ 2.1% 4.2% Oil by Clevenger Method ~___________ 1.2% 8.5% Acidity as mg. KOH/gm. __~_____.____ 9.52 4.41 Refractive index nD 20°/20° ________ 1.4675 1.4675 CNG Kai ce eS HR See aa ey ee ee Similar The results summarized from Table III are: 1. There is twice as much essential oil present in the domestic sample as there is in the imported. 2. The refractive index of the two is identical. 3. The study of the quality of the distillate shows that the fragrance com- pares favorably with standard English Lavender from Mitcham district. Twenty-Eighth and Twenty-Ninth Annual Meetings 17 IV. JUNIPER Juniper is important to our industry. To the best of our know- ledge, it has not yet been cultivated in this country except occasionally for ornamental purposes. However, numerous places of wild growth have been found in various states, and our laboratories tried to cover thor- oughly all these wild sources in order to find the best material suitable for our purpose. It must be said that so far the results of these analyses are satisfactory only to a certain extent; however, there is a very good chance that culti- vation will bring favorable results as it has been possible to select some suitable material from the wild samples. The imported species is the Juniperus communis L., Pinacae and the plant is a perennial. The commercially important part of Juniper is the bluish black, very pulpy fruit or berry having the essential oil in glands situated on the surface of the seeds. The main constituent of Juniper oil is alpha pinene. The other known constituents are cadinene, camphene, terpinene, borneol, and iso-borneol. Because the components are mostly terpenes, Juniper oil is especially un- stable. In the ripe berries the oil is already partly changed into a resin: while if overripe it is resinified almost completely. The oil from green fruit is very unpleasant. Juniper berries find applications in various food products such as candies, packing industries, carbonated beverages, etc. They have pharmaceutical uses and are also used for spice oil and in the liquor industry. TABLE IV Analyses of Juniper Berries (Juniperus communis L., Pinacae) Brcpsy INOS ove ee 169 225 515-1 515-2 ANGE) SS ee 1940 crop 1940 1941 1941 IS OUIMET Vig eas ee Italy USA Wyoming USA Montana USA Montana (J. communis) (J.scopulorum) (J.scopulorum) (J. communis) Moisture by desiccator __-_--____ 10.0% 2.05 % 3.783% 1.324% Moisture by SNOETD, Cea aes ee 13.5% 6.60% 9.982 % 5.454% Total Moisture —~____ 23.5% 8.65 % 13.765 % 6.778 % Oil by Seagram’s Method 2s. e ei 1.22% 2.01% 2.19% 3.81% Oil by Clevenger Methodiy = === sa 1.00% 1.90% 2.35% 4.00% Acidity as mg. KOMYiom = 32 17.3 10.00 7.07 26.11 Refractive index D2 Og 20) Ce 1.4862 1.4816 1.4752 1.4798 Orie eel ee Different Different Different Table IV summarizes the results of analyses of Juniper berries brought from various regions of this country. We will discuss only those that came nearest to our requirements. 18 The Kentucky Academy of Science 1. There seems to be a general tendency for the oil content to be — higher in wild aromatic plants than in cultivated varieties. 2. The quality seems to be almost in reverse proportion to the amount of oil. 3. In some cases, it was possible to approach the pleasant flavor of the control distillate by reducing the concentration of oil from domestic species as much as three or four times. For example, the oil in the second column when diluted in proportion 1:4 had a more pleasant flavor than the standard. In other cases, only by blending with im- ported material could favorable results be obtained. This experience leads us to believe that the main difficulty with the wild domestic Juniper berry is that it is improperly collected. For this reason, we would like to give here a few recommendations on this subject. Juniper shrubs flower in May, but do not ripen their fruit until late in the following year. The berries ripen 2 to 3 years. The amount of oil is most abundant in those berries which have attained their full growth, but are still green. The quality of oil in these green berries is however in- ferior. In overripe berries the oil, although abundant, has been almost totally changed into resins, and only ripe berries have a characteristic frag- rance and should contain from 0.9 to 1.59% of essential oil. The best ripe berries should be at least 1% years old after blooming, have a bluish-black shining color, and should be harvested in the season from October-De- cember. The average size of berries should be 8-10 mm. and they should have no blemishes. If all these conditions are carried out, we believe that domestic Juniper berries will compare very favorably with the imported product. America can produce excellent aromatic plants if a certain prejudice can be overcome and the necessary knowledge acquired. The prejudice we mean is the idea existing in this country that only Europe can pro- duce aromatic plants of good quality and that American soil is not suit- able for such cultivation. The example of Coriander which is already cultivated on a large scale (150 acres) in Kentucky this year seems to bear out our conclusions. Probably this should be true for almost any imported aromatic plant. Our experience in this field would indicate that: 1. Domestically cultivated Coriander, Angelica, and Lavender could totally replace the formerly imported plants. 2. Further research is necessary to improve the quality of domestic Jun- iper and to a lesser degree of Angelica. 3. That there are unusual opportunities for further research in the field of aromatic domestic plants and essential oils. Twenty-Eighth and Twenty-Ninth Annual Meetings 19 THE BRIGHT GREEN BEES OF THE GENERA AGAPOSTE- MON, AUGOCHLORA, AUGOCHLOROPSIS AND AUGOCHLORELLA IN KENTUCKY Harvey B. Lovett University of Louisville The bee family Halictidae contains a group of four genera which are easily distinguished from other members of the family by their bright green or blue-green color. They may be readily separated from the green bees of other families by the strongly curved basal vein in the fore wing and the short tongue. Because of the latter character they are restricted to short tubed or open flowers in their nectar gathering. They do visit many of the longer tubed flowers for pollen, however. They fly from early spring to late fall and are sufficiently numerous to be of considerable assistance in the pollination of many species of plants. These genera have been- recently revised for the United States by the late Grace Sandhouse of the Bureau of Entomology and Plant Quarantine (2,3), whose excellent work has reduced to synonyms many of the super- fluous names which have cluttered up the literature pertaining to these groups. Two other entomologists have made lists and keys for these genera in eastern United States, namely Robertson (1) for Carlinville, Illinois and H. L. Viereck (4) for Connecticut. The resurrection by Sandhouse of older names which have priority has made these papers of Robertson and Viereck difficult to use. In the following keys simple terminology has been used whenever pos- sible but a little knowledge of insect anatomy is necessary to identify in- sects. A good hand lens or a binocular dissection microscope is also indis- pensable. Sandhouse (2 and 3) lists the states from which she has seen specimens but lists only two records from Kentucky, namely, Agapostemon virescens and A. radiatus. The other species therefore appear to be recorded for Kentucky for the first time in this paper. All county locality records refer to Kentucky collections unless otherwise indicated. Key to the females: segments in abdomen, six; joints in antennae, twelve 1. POSTERIOR SURFACE OF PROPODIUM ENCLOSED BY A SHARP- LY DEFINED RIM, DORSAL SURFACE LACKING AN ENCLOSED DISK (AGAPOSTEMON). ; 2. Abdomen black, thorax and head green ..... Agapostemon virescens 2. Abdomen green like the thorax 3. Mesoscutum with well separated punctures of two distinct sizes, large species, 12 mm: long 2) yd ee Agapostemon texanus 3. Mesoscutum with punctures nearly uniform in size and close together 20 The Kentucky Academy of Science 4. Mesoscutum not rugose between punctures, dorsal surface of pro- podium with irregular anastomosing ridges Agapostemon splendens 4. Mesoscutum rugose between punctures, dorsal surface of propodi- um with longitudinal ridges, small under 10 mm. Agapostemon radiatus 1. POSTERIOR SURFACE OF PROPODIUM NOT ENCLOSED BY A COMPLETE RIM, DORSAL SURFACE WITH AN ENCLOSED DISK. y. First and second tergites with an apical fringe of bristles, inner spur of hind tibia with a few long teeth (AUGOCHLOROPSIS) 6. Vertex elevated above ocelli, humeral angles of pronotum forming wing-like projections, 12 to 13 mm. ..... Augochloropsis caerulea 6. Ocelli on highest part of head, humeral angles of pronotum forming sharp ridges only,’ 10 mm: long). 4 ...0. Augochloropsis cuprea §. Fringe of bristles absent, innerspur of hind tibia merely serrate 7. First sternite with prominent median ridge, mandibles with two nearly equal teeth (AUGOCHLORA) ........ Augochlora pura 7. First sternite without ridge, mandibles with one large and one small tooth (AUGOCHLORELLA) 8. Disk not ridged to posterior margin, ridges very irregular, meso- scutum closely and uniformly punctured .. Augochlorella aurata 8. Disk ridged to posterior margin, ridges longitudinal and regular, mesoscutum rugose-punctate anteriorly 9. Disk with about 24 to 30 nearly straight prominent POSES EMS. apc kile don cused aan eS Re ae Augochlorella striata 9. Disk with 36 or more fine, slightly irregular ICES 1 eae the NaN ce ae ee Augochlorella gratiosa Key to the males: segments in abdomen, seven; joints in antennae, thirteen . ABDOMEN BANDED WITH ALTERNATE Eo AND DARK BANDS (AGAPOSTEMON) 2. Dark bands on abdomen strongly tinged with ice green, dorsal sur- face of propodium with triangle in center. ...Agapostemon texanus _ 2. Dark bands not tinged with blue-green, no ‘triangle on propodium 3. Hind femur so strongly swollen that its width is more than one- inalgats@lenechiiy a. eater eet sets me tetas Agapostemon splendens 3. Hind femur less than one-third its length 4. Apical margin of 4th sternite with a green spot, front and mid- dle trochanters largely yellow ......... Agapostemon radiatus — 4. No green spot on 4th sternite, trochanters black ...... Agapostemon virescens 1. ABDOMEN GREEN OR BLUE-GREEN, NOT BANDED WITH YELLOW 5. First and second abdominal tergites with apical fringe of bristles, tegulae large with blunt caudal borders (AUGOCHLOROPSIS) 6. Vertex elevated above ocelli, tarsi dark brown Augochloropsis caerulea 6. Ocelli on highest part of head, tarsi yellow. .Augochloropsis cuprea Twenty-Eighth and Twenty-Ninth Annual Meetings Di §. First and second abdominal tergites without apical fringe of bristles, tegulae small and rounded 7. Second flagellar joint longer than first, apex of radial cell truncate with a tiny vein attached (AUGOCHLORA) .. Augochlora pura 7. First and second joints nearly equal, apex of radial cell pointed (AUGOCHLORELLA) 8. Disk of propodium not ridged to posterior margin, mesoscutum closely and uniformly punctured ......... Augochlorella aurata 8. Disk of propodium ridged to posterior margin, mesoscutum later- ally and anteriorly rugose 9. Hairs at base of hind basitarsus more than half its length, apical margin of 4th sternite with deep indentation........... Augochlorella gratiosa 9. Hair at base of hind basitarsus less than half its length, apical margins of 4th sternite with shallow indentation ........ Augochlorella striata TERMINOLOGY. Dorsal plates on thorax from the anterior end are: PRONOTUM, MESOSCUTUM, MESOSCUTELLUM, METANOTUM; PROPODIUM: the last segment of thorax (really derived from abdomen), the dorsal surface of which usually has an enclosed disk; TERGITES: plates on the dorsal side of abdomen; STERNITES: plates on ventral side of abdo- men; TEGULAE: small scales at base of wings; RADIAL CELL: the cell on anterior margin of fore wing just distal to stigma (black spot); BASITAR- SUS: first segment of tarsus; VERTEX: top of head; TROCHANTERS: second segment of legs; FLAGELLUM: the eleven (female) or twelve (male) distal joints of antennae. AGAPOSTEMON— Guerin Agapostemon has the posterior surface of the propodium enclosed by a conspicuous rim but the dorsal surface lacks the enclosed disk, and the tibia of the hind leg is as long as the tarsus. The sexes vary greatly, the females usually having green abdomens (black in one eastern species) and the males having yellow bands alternating with black ones. 1. Agapostemon virescens (Fabricius). Females: the only eastern species with black abdomens. Males: the yellow and black banded abdomens lack the greenish tinge entirely, hind femur less than one-third as long as broad, trochanters black. A fairly common species in Kentucky. Has been collected on Melilotus alba, Pentstemon canescens, Hibiscus syriacus; from June 9 to Aug. 5; Jefferson Co. 2. Agapostemon radiatus (Say). Females: abdomen green, plate on dorsal side of mesothorax (mesoscutum) with uniform punctures, rugose be- tween punctures. Males: bands on tergites of abdomen not tinged with green, but a green spot on margin of 4th sternite, front and middle trochanters yellow, hind femur more than three times as long as broad. A fairly com- mon species. Has been collected on Teucrium canadense, Pentstemon cane- 22 The Kentucky Academy of Science scens, Camassia esculenta, Dianthera americana and cultivated Weigelia; from May 5 to Aug. 10; Jefferson, Warren and Powell Cos. 3. Agapostemon texanus (Cresson). Females: abdomen green, meso- scutum with well defined punctures of two distinct sizes. Males: dark bands on abdomen strongly tinged with blue-green, dorsal surface of propodium with faint triangle in center. This species is confined in the eastern part of North America to the northern states, although in the west it extends as far south as Texas. It should occur as far south as Kentucky, although no speci- mens have been obtained as yet. I have two males collected at Rimersburg, Penn. on Mentha piperita. 4. Agapostemon splendens (Lepeletier). | Females: abdomen green, mesoscutum with punctures nearly uniform in size and not rugose. Males: easily recognized by the swollen hind femur which is more than half as broad as long. The range covers the United States east of the Rockies. No speci- mens have been obtained in Kentucky. AUGOCHLORA—F. Smith Apex of radial cell truncate with appendiculate vein, the posterior sur- face of propodium not enclosed by a rim, no apical fringe of bristles on ter- gites one and two, hind tibia longer than tarsus. Females: first sternite with a high, median ridge, mandibles with two nearly equal teeth, inner spur on hind leg finely serrate. Males: second flagellar joint longer than first. As redefined by Sandhouse (2), this genus now contains only one species in the United States east of Texas. 1. Augochlora pura (Say). A bright green bee about 9 mm. in length, with the characters of the genus. Has been taken on Teucrium canadense, Camassia esculenta; May § to July 11; Jefferson and Oldham Cos. AUGOCHLORELLA—Sandhouse Apex of radial cell pointed, tegulae small and oval, posterior surface of propodium not enclosed by a rim, no apical fringe of bristles on tergites one and two, hind tibia longer than tarsus. Females: mandible with one large and one small tooth, first sternite without median ridge, inner spur of hind tibia finely serrate. Males: first and second flagellar joints nearly equal. This genus was erected by Sandhouse in 1937 and so closely re- sembles Augochlora that it might well be considered merely a subgenus. 1. Augochlorella striata (Provancher). Females: disk of propodium ridged with 24 to 30 nearly straight prominent ridges, mesoscutum rugose-punctured on anterior margins. Males: apical margin of 4th sternite with a shallow indentation. This species has been known as Augochlora confusa Robertson until Sandhouse discovered that it was identical with the species described earlier by Provancher. Has been taken on Camassia esculenta, Asclepias tuberosa, A. syriaca, Erigeron philadelphicus, Houstonia coerulea, Cunila Twenty-Eighth and Twenty-Ninth Annual Meetings 23 originoides, and Monarda fistulosa (sucking through holes bitten by some insect in tube), Companula americana, and Melilotus alba; from May 5 to September 19; Jefferson and Oldham Counties. 2. Augochlorella gratiosa (Smith). Female: disk of propodium with 36 or more fine rather irregular ridges running to the posterior end, mesoscu- tum rugose-punctate. Males: apical margin of fourth sternite with a deep indentation. This species is difficult to distinguish from the preceding as there is considerable variation in the regularity and number of ridges on the disk. Taken on Asclepias syriaca, Cunila originoides, Erigeron phila- delphicus, and Teucrinm canadense; May 11 to Sept. 19; Jefferson Co. 3. Augochlorella anrata (Smith). Both sexes may be distinguished by the irregular striae on disk which do not reach to the posterior margin, and by the lack of rugosity on the mesoscutum. Azgochlora similis Robt. is a synonym. Taken on Asclepias tuberosa and Claytonia virginica; from April 29 to June 23; Jefferson Co. AUGOCHLOROPSIS—Cockerell This is a well defined genus, distinguished by an apical fringe ‘of bristles on the first and second abdominal tergites, the large tegulae with blunt caudal margins, and the lack of a raised rim on the posterior surface of the propodium. Females: Spur of hind leg with a few long teeth, mandible with one large and one small tooth. Only two species are known in the United States both of which occur in Kentucky.. 1. Augochloropsis caerulea (Ashmead). Both sexes have vertex elevated above ocelli, antero-lateral margins of pronotum sharply projecting, bluish tinge, large (12 to 13 mm.). Has been taken on Apocynum cannibinum, Asclepias incarnata, A. tuberosa, Dianthera americana, Melilotus alba, and Solanum; from June 3 to July 19; Jefferson, Todd, and Warren Cos. 2. Augochloropsis cuprea (Smith). Females and males: ocelli on top of vertex, antero-lateral margins of pronotum form shallow ridges, golden- green, smaller (9 to 10 mm). Has been taken on Camassia esculenta, and Melilotus alba; from May 5 to July 19; Jefferson and Oldham Cos. Literature Cited 1. Robertson, Charles, 1902. Synopsis of the Halictinae. Can. Entom., 4: 243-250. 2. Sandhouse, Grace A., 1936. The bees of the genus Agapostemon oc- curring in the United States. J. Wash. Acad. of Sci., 26: 70-83. 3. Sandhouse, Grace A., 1937. The bees of the genera Augochlora, Augo- chloropsis, and Augochlorella occurring in the United States. J. Wash. Acad. of Sci., 27: 65-79: 4. Viereck, H. L., 1916. Guide to the Insects of Connecticut. Halictidae pp. 699-707. 24 The Kentucky Academy of Science MODIFICATION OF DEVELOPMENT IN BRUCHID (Coleoptera) EMBRYOS BY TREATMENT WITH AMINO ACIDS ALFRED BRAUER Department of Zoology University of Kentucky Of primary importance in metabolism are the so-called fundamental amino acids. They constitute not only structural blocks of a protoplasm but actively share in its operations in the sense that their molecules are continually changing and renewing themselves (5). The enzyme system of a protoplasm, at the basis of which is the gene complex, is constantly acting upon, or interacting with the amino acids for the liberation of energy as variously expressed throughout a life cycle. In that period of the cycle immediately following fertilization of the ovum it is expressed in rapid cell division, development and differentiation. This being the case, it might now be assumed that the interactions be- tween oxidative enzymes and amino acids and consequently the rate of embryonic development can be augmented by enhancing the availability of the fundamental amino acids during early development, and that develop- ment might be progressively modified thereby. It is the purpose of this investigation to determine whether or not this is the case. Methods Eggs of the so-called pea weevil, Bruchus quadrimaculatus Fabricius, deposited on sterilized peas over 30-minute intervals were removed from the peas and at the desired age were immersed and incubated in solutions ef amino acids. After numerous trials the time of treatment was estab- lished at 24 hours, after which the eggs were removed from the solution, washed and transferred to Ringer’s solution, and incubated in this until their total age was 48 to 52 hours. The amino acids used were chlorides of d-l tyrosine, | beta-phenylalanine, and cysteine in concentrations ranging from 1/2000 to 1/200,000 mol. Hydrogen-ion concentration was originally not controlled. For tyrosine and phenylalanine this was pH 6.4 and 6.5 respectively, and known to be in the range of tolerance for the embryos. Cysteine hydrochloride being more acid, pH 6.0, was diluted with Ringer’s solution and there-. by given a pH of 7.2. Controls were then run with the two former acids dissolved in Ringer’s solution but without detectable differences in reaction of the embryos. After incubation, the eggs were fixed, stained and embedded in celloidin. In this they were cleared for entire study. Afterward if desired, they were prepared for histological study by the celloidin-paraffin combination method. Twenty-Eighth and Twenty-Ninth Annual Meetings 25 Modifications after Tyrosine Treatment Practically none of the treated eggs were entirely normal, though it is dificult to set a criterion for normality. They may be classified as 68 percent nearly normal, 17 percent gross malformation, 15 percent, no de- velopment. The more completely normal are only smaller than the average embryos of untreated eggs. Symmetry and general appearance of the embryos are otherwise normal. A minor deformity shown by many embryos is that the body is bent laterally to either side, and sometimes it is curved upward from the ventral to the dorsal side of the egg. Failure of the caudal plate, prospective abdomen, to contract entirely by the 50-hour stage as it does normally by 36 hours, is a common fault though this is hardly peculiar to tyrosine treated eggs. It may indicate retarded development. (Fig. 1). From such embryos the transition is toward those which show most or all of the abnormalities mentioned, and in addition are more deeply seg- mented. Head lobes may be small, and mandibular rudiments asymmetri- cal. Such embryos usually stain more intensely with carmine, so that they appear dark. Finally there appear contorted, twisted or very abnormally flexed em- bryos. In these there is lack of uniformity as to the mode of flexure. Sev- eral appear as though there were a mild tendency toward thoracic duplicity, but nowhere in the tyrosine series were embryos found which definitely showed duplicity of any sort (Figs. 2 and 3). Sectioned, undeveloped eggs show first, that the cleavage nuclei are en- larged. They have a reticular structure internally, but the chromatin appears scanty. In some eggs it appears as though nuclei had failed to divide regularly. Second, the cytoplasmic cortex of the egg is more or less irregular and thinner than normal, while in a few the cortex seems to be entirely lack- ing. Cleavage nuclei may either have failed to reach the cortex, or they may have reached and entered it. In the former, division and peripheral movement of the cell appear to have ceased. In the latter, failure to de- velop appears not to have been the fault of the nuclei, since these have entered the cortex and here have proliferated freely. Proliferation appears rather to have taken place without organization into either a blastoderm of distinct cellular units or into an embryonic plate. Sectioned embryos show complete organization of structures and tissues. Mitotic figures are numerous especially where localized development occurs. Injured nuclei, instead of being enlarged as in cleavage stages, are smaller than normal especially in that part of the embryonic plate from which the ganglionic rudiments of the nervous system form. Here they are found in groups more or less surrounded by normal neuroblasts. Considerable recovery has taken place previous to this differentiation. Fig. 1. A nearly normal embryo of 52 hours. Treated for 24 hours in 1/12,000 mol. tyrosine. Fig.2. Embryo of 52 hours, treatment same as No. 1. Distorted, deeply segmented, dwarfed, no ventral ganglia developed. Fig. 3. Embryo of 48 hours. Treated with 1/2000 mol. Phenylalanine. Embryo twisted and contorted. This form also appears often in the tyrosine series. Fig. 4. Embryo of 52 hours. Treated with 1/12,000 mol. Phenylalanine. Unintegrated development of three areas. Fig. 5. Embryo of 50 hours. Treated with 1/12,000 mol. Phenylalanine. Partial duplicity of thoracic region. Fig. 6. Embryo from the same egg lot as preceding. Double head, prothorax and lower thorax. Conjoined in mid-thorax and lower abdomen. Fig. 7. Embryo from same series as 5 and 6. Anteriorly the same as preceding. Duplicity of abdomen in which one part has again partially reduplicated itself. Fig. 8. Embryo of 48 hours treated for 24 hours with 1/10,000 mol. cysteine. Body por- tions are poorly. integrated. Embryo has developed on side of the egg. b Twenty-Eighth and Twenty-Ninth Annual Meetings 27 Modifications after Phenylalanine Treatment Here modifications are definitely more pronounced than in those of the tyrosine series. Eighteen percent of the eggs failed to develop; 28 percent gave gross malformation; 52 percent are fairly normal. Dwarfed, contorted and twisted embryos appear in nearly every treated egg lot. Since many of the abnormal embryos are similar to those of the tyrosine series, only those peculiar to phenylalanine treatment will be de- scribed. These are embryos showing several forms of unintegrated de- velopment. First, complete unintegrated development of several embryonic rudiments is found in several eggs. Isolated parts appear anteriorly, later- ally and posteriorly on the ventral egg surface (Fig. 4). Whether or not these represent special embryonic parts cannot be determined, but it is likely that they are not sufficiently organized to show differentiation of head or abdominal portions. Second, unintegrated axial development, or partial twinning is shown by numerous embryos of different lots. The first of these is microcephalic and divided thoracically but conjoined in head and abdomen. The caudal plate of the embryo has not contracted (Fig. 5). A second case represents lack of separation in a portion of the thorax and abdomen and duplicity of head and a portion of the thorax (Fig. 6). A third is conjoined in the mid-thorax only. Anterior thorax and head have separated into two asymmetrical portions. There is a divided abdomen in which the left abdominal part has again divided. In other words it is triplicity of the abdomen (Fig. 7). Single but contorted embryos differ from those of the tyrosine series in at least one respect, namely, they are much more elongate. In consequence the distortions are emphasized, probably because the entire embryonic plate is more seriously affected and because a contraction of the embryonic plate from the earlier elongate phase has not taken place as it normally does at 24 to 36 hours of development. The embryo represents an arrested earlier developmental stage. Modifications after Cysteine Treatment Of this series 16 percent failed to develop, 39 percent gave grossly mal- formed embryos and 45 percent are nearly normal. From this it appears that cysteine is as toxic as phenylalanine although this may not actually be the case. The gross malformations are more like those of the tyrosine series in that the embryos are contorted rather than unintegrated. There are no cases of duplicity in this series. One of the most extremely malformed embryos has developed laterally around the surface of the egg yolk. While it has sev- eral distinct segments the relationship of these to definite body aréas can not be determined. Secondary segmentation is indefinite and head parts have failed to differentiate (Fig. 8). A majority of the treated eggs have a more normal form for their age 28 The Kentucky Academy of Science but are dwarfed and somewhat twisted. A failure of the abdominal plate to contract after the head and thorax have undergone greater differenti- ation is a common fault. Discussion — Conclusion The amino acids | tyrosine, dl beta--phenylalanine and cysteine definitely exhibit inhibitory toxic action on the developing bruchid embryo. Like toxic substances generally, their inhibitory action is expressed differentially from centers of high metabolic activity to regions of lower rates. In the very early insect egg the cleavage cells are especially susceptible, while slightly later (3 to 6 hours after oviposition) the antero-ventral cortical cytoplasm becomes most susceptible. This differential susceptibility to- gether with differential recovery or acclimatization can account for all of the reconstituted forms here exhibited (4) (2). The effects of tyrosine and cysteine appear to be exhibited first on the susceptible cleavage cells in the complete or partial arrest of the cleavage process and then to partial arrest of activity in the cortical cytoplasm. In no cases has this secondary arrest brought about the complete isolation of potential secondary centers which would have resulted in unintegrated developments and in duplicities. Phenylalanine on the other hand may completely arrest the early integration center of the antero-ventral cyto- plasm to produce duplicities and even triplicities. After treatment with lower concentrations of the acids less complete arrest and more complete differential recovery is the rule, yet peculiar modifications may result. The method of action or interaction between the acids and the proto- plasm is not known and only suggestions can be offered. The optic activity of the amino acids and the capacity of protoplasms to produce both dextro- and laevo- isomers in these molecules, together with the differential actions of the asymmetric molecules on the cytoplasmic mechanism may be a possibility (1). On the other hand the effect of the acids on the oxidative mechanism is not precluded. The supplied amino acids, acted upon by oxidases of the cells or of the egg cortex, possibly more readily than those which are constitutional to the cytoplasm, actually inhibit normal intra- cellular oxidations and thereby produce a suffocation effect (6). References 1. Bate, Sir Hardy William, 1934. Abraham Flexner Lecture Series, No. 2, p. 28. Williams & Wilkins, Baltimore. 2. Brauer, Alfred, 1938. Physiol. Zool., 11: 249-266. 3. Brauer, Alfred and Taylor, Albert Cecil, 1936. Jour. Exp. Zool., TBS WAS Ihe 4. Child, C. M. 1940. Patterns and Problems of Development, Univ. of Chicago Press. 5. Schoenheimer, R. 1940. Physiol. Rev., 20: 218. 6. Van Slyke, Donald D. 1942. Science, 95: 259-263. a a Oe Twenty-Eighth and Twenty-Ninth Annual Meetings 29 ORIGIN AND DEVELOPMENT OF PRICKLES OF ARALIA SPINOSA P. A. Davies University of Louisville The great development of sclerenchyma in aerial parts of many plants is associated with the development of prickles or spines (thorns). These sharp pointed structures, though not lacking in other regions, are characteristic of many xerophytes of desert and steppes. A few mesophytes produce these sclerenchyma structures to a greater degree than many xerophytes. Aralia spinosa (Hercules’ club) is a mesophyte which produces an abundance of sharp pointed sclerenchyma protuberances particularly on the first and second season’s stem growth. Because the literature is not clear as to the definite terminology to be used in describing these sclerenchyma protuberances, the classification by Maout and Decaisne will be followed (1). According to them, the scleren- chyma protuberances are grouped into two types: spines or thorns which result from reduced structures, i.e., stems, leaves and flowers, and contain conducting tissue, and prickles which result from out-growths of epidermis and cortex and contain no fundamental tissue. The sharp-pointed scleren- chyma growths in Aralia spinosa belong to the latter group. The prickles appear on the new growth in the spring and continue to elongate until the wood ripens in early summer. They are not evenly dis- tributed over the stem; the heaviest concentration occurs during the first season’s growth (nearest the ground) and each succeeding year’s growth produces fewer prickles. Although the prickles are scattered without def- inite arrangement over the stem, they are more abundant at the leaf bases. The first appearance of the prickles is a localized outward arching of the cortex and epidermis. Microscopic examinations of prepared slides at this early stage of development show a mass of small cortical cells. As the cells in the localized mass continue to enlarge, those in the outer area push the center cells into a plane in which their long axes are at right angles to the stem. The gradual elongation of these vertical cells with the covering epi- dermis, form the prickle. As there is no vascular supply to the prickle, the growth is limited in proportion to the diameter of the initial arching cell- mass. The length of the mature prickle varies from 1 to 15 millimeters, with an average of 7.08 millimeters. The prickle hardens through lignification of the cell structures. The cells at the apex lignify first, followed by a progressive lignification inward and toward the base. The lignification is rapid, and by the time growth ceases, a sharp-pointed, tough prickle is formed. 1. Maout, E., and J. Decaisne, 1876. A General System of Botany, pp. 17, 26, 138. Longmans, Green and Co. 30 The Kentucky Academy of Science ABSTRACTS A review of Kentucky botanical history. H. T. SHackLeTTE, Depart- ment of Botany, University of Kentucky.—The serious study of plants of — Kentucky was first attempted by Andre Michaux in 1793, who botanized in this region while carrying out a political mission for the French Goy- ernment. His son, Francois Andre Michaux, collected in this area in the early part of the nineteenth century, and was contemporary with McMurtrie of Louisville and the Transylvania University group of botanists, con- sisting of Rafinesque, Short, Peter and Griswold. The works of these early botanists constitute the only major effort toward a comprehensive listing of Kentucky plants. With the end of this period, the study of this science was largely neglected until the botanical works of the Kentucky Geolog- ical Survey were issued in the latter half of the century. A period then followed (which has persisted to the present time) of sporadic isolated works, consisting of five county floras, several ecological papers, lists of particular groups of plants, and a great number of isolated references in taxonomic monographs to Kentucky plants. It may be said that the flora of this state is known to the majority of botanists of both this country and Europe from the collections which Dr. C. W. Short and his associates made over a century ago. A mimeographed list entitled ‘‘A Preliminary Bibliography of Kentucky Botany” September, 1940, has been compiled. It contains 370 references. A second list entitled “First Supplement to a Preliminary Bibliography of Kentucky Botany,” April, 1941, expanded the references to 441 titles. These publications have been disco buted to all workers in Kentucky known to be interested in this project. The earliest movements of the Bruchid embryo and remarks concerning the neuro-motor mechanism development of corresponding stages. E. BRUCE NeEwELL, Department of Zoology, University of Kentucky.—A study has been made of the neuro-motor development of Bruchid (Coleoptera) em- bryos, and an attempt is made to correlate this development with the earliest movements. The earliest movements. begin between approximately 68 and 72 hours of age and consist primarily of ventro-lateral contractions in the thoracic region. The very first movements are wholly spontaneous but within a few hours the embryos respond to a stimulation of the an- terior end. As age increases, the movements become evident in the more posterior regions of the body, and by about 75 hours they begin to assume the form of general bodily contractions, and the embryo will withdraw quickly from the point of stimulation. That the early behavior is not dependent upon simple ‘“‘protoplasmic conduction” is evidenced by the presence of nerve fibers, commissural connections, and the formation of muscle fibers on the ventro-lateral side. As the movements progress in a cephalo-caudal direction, they are preceded by the development of both receptor and motor elements. Twenty-Eighth and Twenty-Ninth Annual Meetings 31 Spectrochemical Analysis.* Epwin S. Honce, Agricultural Experiment Station, University of Kentucky.—In recent years the use of the spectro- graph for qualitative and quantitative chemical analysis has received a great deal of attention. This paper deals with some of the instruments, techniques and results of the spectrographic methods to research and routine control. The use of different types of instrument and-light sources for biological, ferrous and non-ferrous analysis is described. Many industries are using spectrographic methods for routine control and several of these techniques are described. In consideration of the time saved and accuracy attained these methods are much superior to the usual chemical procedures. The application of similar methods is equally important to research prob- lems. The results of several such investigations are given together with the accuracy and sensitivity obtained. This offers a solution to many problems which would be difficult or impossible to solve otherwise. A spectrographic study of the tissues of wobbler horses. Eyow1tn S. HopceE, W. S. Hopexiss and B. J. Errtncton, Agricultural Experiment Station, University of Kentucky.—For some time, particularly in Kentucky, it has been reported that a condition characterized by a bilateral incoordination of the hind limbs exists in horses. Certain symptoms in common with similar diseases in other animals led to the belief that this condition may be due to a mineral deficiency. With this in mind the ribs, vertebrae, spinal cord, and livers of some diseased and normal horses have been studied spectrographically. Examination of the data shows that about 20 elements are present. An approximation technique was used to determine the amount of certain elements present. Others were estimated as present in trace and micro quantities. This data is compiled in a table to show the occurrence of elements in relation to whether the horse was a normal one or a wobbler. In this preliminary survey no significant correlation could be found in the mineral composition of tissues of either normal or diseased animals. Sources of minor element contamination in sand culture experiments with plants. R. H. Haceman and J. S. McHarevue, Agricultural Experiment Station, University of Kentucky.—The establishment of nutritional defi- ciencies of the minor elements in plants grown in cultural media is com- plicated by contamination in chemicals, water, sand and containers. In recent experiments performed at the Department of Chemistry, Ken- tucky Agricultural Experiment Station, chemicals were purified by a com- bination of the Steinberg procedure and a dithizone purification. Pure water is obtained by distillation in a still having a special quartz condenser. Pure sand is washed with acid to remove all minor elements. Glassware or well-glazed porcelain pots are satisfactory containers. Lettuce, tomato and Swiss chard plants developed severe boron defi- * Material published in full in: J. S. McHargue and E. S. Hodge, ‘‘Spectrography in Agri- cultural Research,’ J. Assoc. Off. Agr. Chemists, Vol. 25 (1942). 52 The Kentucky Academy of Science ciencies when grown in a sand media and supplied with an unpurified nutrient solution made from reagent grade chemicals. Tomato plants were the only plants that developed a manganese deficiency when grown in this media. It is apparent that nutritional deficiencies of the trace elements may be | produced in vegetables only by careful purification of chemicals, sand, water and containers. Determination of moisture on tobacco (oven method vs. vacuum desic- cator method). C. W. WoopMansEE, Karu E. Rapp and J. S. McHarcugE, © Agricultural Experiment Station, University of Kentucky.—For many years — che cleaeamingion of melee on agricultural products has been a subject — of controversy. Perhaps the most reliable source of reference on this sub- — ject is the Official and Tentative Methods of Analysis of the Association of Official Agricultural Chemists; and yet, in it one finds no definite'method — for tobacco. , A study was made to ‘compare the moisture determined by drying in an | oven at 100°-105° C. and in a vacuum desiccator over sulphuric acid at room temperature for various intervals of time. Graphs of the data plot- ting percent moisture against time of drying shows the vacuum desiccator | method to give a better indication of the true moisture present in tobacco. ~ This conclusion is made because it is believed that some of the constituents of tobacco other than moisture are volatilized by the oven method, and thus would not give satisfactory results. * Published in full in J. Assoc. Off. Agr. Chemists, Vol. 25 (Feb. 1942). TRANSACTIONS OFTHE. 7. KENTUCKY ACADEMY OF SCIENCE (Quarterly Series) : AFFILIATED WITH THE A.A.A.S. Bes VOLUME 10 a (Double Issue) a NUMBER 3, DECEMBER 1942 A NUMBER 4, MARCH 1943 fg TWENTY-EIGHTH AND TWENTY-NINTH a ANNUAL MEETINGS | 1941-1942 CONTENTS u Page The Semimicro Determination of Magnesium as the Quinolate ¥ Using the Colorimetric Ferric Chloride Method f M. E. Weeks ae eee eRe Todds fo 5 aMbee sak i oath ol 2 ee 33 Studies on Human Parasites in Kentucky. JI. Preliminary Note on the 5 Occurrence of Endamoeba histolytica and related species in a Group of College Students Davidwiwehand, Lincigome ia iti oc. 2 41 The Measurement of Relative Intensities of Soft X-Rays by the Photo- Al _ graphic Method. % i@lramesales Owens” MAGE es TON it Lak Pe N eae ae 43 i 2 ESSAYS as LC) APN RS a i A PMO 45 OLAS UOTE ea eel UN |. De EG DR eM 49 é } j = TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE 4 irr ie 4 . EDITORIAL STAFF Rt x 4 Dayrence Baker’ io 0 us Berea College ____ Psychology and Philosophy We Cook. ois shot 8 sy) Gentrey Callegey i!" ve ae eas ie H. B. Lovell _________-_ University of Louisville _..--__ Biology | WG.) McFarlan! 8.0) Wl University of Kentucky ______.___ Geology | ward) Gr Sumpter) 2202 Western State Teachers GolleeG __ Chemistry 5 fevenratoWl Boye cs iy ice aliaaie Nene La a University of) Kentucky: 4) 2 2a Physics Johns Kurperi) ass sak ui University of Kentucky, Managing Editor 4 >. Manuscripts. The Transactions must be limited to the proceedings of the annual meetings of the Kentucky Academy of Science and to original manuscripts pertaining to science. Manuscripts are subject to the ap-~ proval of the Editorial Staff and may be submitted to the Editor of the subject covered or to the Managing Editor. Exrra-Cost Features. ‘The extra cost of special features such as cuts, graphs, tables, etc., above the text-run price per page must be borne by the — contributor. The Editorial Staff will advise contributors concerning the ~ extra cost of features upon receipt of manuscript. Illustrations to be in- © eluded in an article should accompany the manuscript if possible, or, if sent — in separate package should be properly labeled as to the article in which they - are to occur. : ; Proor. Galley proof will be sent for approval of contributors. The proof should be returned piompely to the Berea College Press, Berea, Ken- — tucky. . Reprints. Reprints are furnished at publisher’s prices by negotiating directly with Berea College Press. Price quotations on reprints are sub- mitted with the proof. Orders for reprints should accompany the proof to the Berea College Press, Berea, Kentucky. SUBSCRIPTION Rates. The Transactions is sent without additional ex- — pense to all members of the Kentucky Academy of Science who are not in © arrears for annual dues. The annual-subscription rate for non-members — is $2.00 in the United States and Canada, $2.50 in foreign countries; single numbers 75 cents. One volume of four numbers appears each Academy — fiscal year. ' | BusINEss CORRESPONDENCE. Remittances and correspondence concern— — _ ing subscriptions, extra costs, and other financial matters except reprints should be addressed to John Kuiper, University of Kentucky, Lexington, = > He ey / 323549 7 | Twenty-Eighth and Twenty-Ninth Annual Meetings 33 THE SEMIMICRO-DETERMINATION OF MAGNESIUM AS THE QUINOLATE USING THE COLORIMETRIC FERRIC CHLORIDE METHOD. *7+ M. E. WeEKs aNnpD Jack R. Topp Agricultural Experiment Station University of Kentucky In working with plant analyses for several years it has been found ad- visable in many cases to change from the slower macro methods to the newer semi-micro methods to save time and reagents. Much time and effort has been expended obtaining the required accuracy along with speed. In some cases the ordinary methods did not prove satisfactory when applied to the small quantities of an element encountered in semi-micro analyses; ‘so new or modified methods had to be substituted. One of these cases was in the determination of magnesium. Several of the semi-micro procedures for this element were tried wherein the magnesium was precipitated as the ammonium phosphate, but the reproducibility of the results was always poor. In duplicate determinations on the same sample it was difficult to obtain good agreement and quite often these duplicates could not be made to check within ten per cent. In attempting to improve the accuracy of the magnesium determination, it was decided to turn to precipitation with 8-hydroxyquinoline since a number of recent authors had reported good quantitative results with this reagent (1) (2) (3) (5). _ The first method tried was that of Wolff (7) because it was simple and readily applicable to biological materials. It gave fair results with standard solutions but when applied to actual plant samples and soil extracts, it was usually necessary to make three determinations to get two that gave good agreement. This naturally left some doubt as to the method’s reliability. ‘Consequently it was decided to study the reaction between 8-hydroxyquin- oline and magnesium in standard synthetic solutions to determine the best ‘conditions for precipitation and estimation. * The investigation reported in this paper is in connection with a project of the Kentucky Agricultural Experiment Station and is published by permission of the Director. 7 Eprror’s Note. A special committee selected this paper as the best read at the 1942 meeting. The authors were awarded the King Prize of $50. for the year 1941-1942. 34 HLONA1 AAVM SNOYOINITIIWN NI The Kentucky Academy of Science TRANSMISSION (PERCENT) aS o Oy) o) O Oo O¢€ OOvV OOS 009 OOL ww GL | a] Fj CeCe oe Fa | 29 TET ao ee my ff T CEE heph RS x HH E PCEECEEE = ri co } [o) a cS) lon) [@) +H o4 a 38 fal ee oO 7 38 oe -00 Twenty-Eighth and Twenty-Ninth Annual Meetings E 8 + . nis | Ee auaaeH nian GIL [ [ moa Cel il e if PHT EERRE- PERE Cee Eee eee ne PS 8 I Ht f ot ; tH t pay fin let cu ae 1 maa i Ou IL 8 nan i tt SE ; eat Laer HeunIEGrEaE led Wie [ eT TORANUEL | t nani Yet 36 The Kentucky Academy of Science Estimation of Quinolate. Of the variously proposed ways for estimating the quinolate, two have proved satisfactory in our work. _ One involves the use of a ferric chloride- acetic acid reagent to develop the green color which is measured with a *“Cenco-Sheard-Spectrophotolometer.”” The other is the titration method using ammonium hexanitrato cerate in 2 molar perchloric acid proposed by Nielsen (5). The reaction between 8- hydroxyquinoline and ferric ions was first ap- plied to the quantitative estimation of iron in 1935 by Lavollay (4). It was found that when the logarithm of the concentration of 8-hydroxyquin- oline was plotted against percentage transmission a straight line resulted and the reaction obeyed Beer’s law. It was therefore adopted for further study. This principle was first applied in estimating magnesium hydroxyquinolate in 1938 by Wolff (7). A transmission curve (Fig. 1) was constructed and maximum transmission was found at a wave length 520 millimicrons with maximum absorption from 560 to 660 millimicrons. It was con- venient to use 560 millimicrons for future readings because at this point transmission of the green color of iron hydroxyquinolate was at a minimum and the yellow of FeCl; nearly at a maximum. Figure 2 gives the con- centration curve for magnesium hydroxyquinolate. The green color of ferric hydroxyquinolate was found to be very stable, keeping without change for a period of several days. The effect of the iron and acetic acid on the color intensity was de- termined by varying the concentration of each in the reagent. A study of absorption data showed that the concentration of ferric chloride had to be controlled within rather narrow limits but that of acetic acid could vary considerably without effect on the results. Precipitation of Magnesium. The precipitation of magnesium with 8-hydroxyquinoline is quantitative even in very dilute solutions and the composition of the precipitate is con- stant (2). Precipitation occurs only in strongly alkaline solutions and since a considerable number of other ions are precipitated under these condi- tions, they must be removed by previous separations. The work of Redmond and Bright (6) has shown that precipitation of magnesium is brought about most rapidly by making the solution strongly alkaline before adding the quinolate reagent. Most procedures, however, call for adding the hydroxyquinoline to a neutral or only slightly acid eleion containing ammonium chloride and then adding the excess ammonium hydroxide. This was the procedure used in this study. It has also been found that a certain excess of quinolate is necessary for the best results. In our early work it was found necessary to use one-half ml. of a two per cent solution of 8-hydroxyquinoline to precipitate magnesium in amounts ranging up to 0.7 Twenty-Eighth and Twenty-Ninth Annual Meetings 37 milligram, and one ml. for greater amounts. In both cases this amounts to a considerable excess but the effect of varying the amount was further checked by experiment and the results are given below. Mg. Magnesium MI.1% Mg. Magnesium Percent used quinolate recovered recovered A415 1 42 101 415 2 Aik 101 415 4 Als 100 It would appear from the above data that there is no effect dut to increas- ing the amount of 8-hydroxyquinoline used in precipitation. However, it seemed that the greater excess caused a slightly more granular precipitate that was easier to centrifuge down. A study was made of the time necessary to give complete precipitation of magnesium hydroxyquinolate. A fairly definite time period was found to be necessary not only to give complete precipitation, but also to maintain the most satisfactory conditions during precipitation. In all cases the solutions were heated on a water bath at about 75° Centigrade after which they were allowed to stand in the cold for periods ranging from twenty minutes to twenty-four hours or longer. If the solutions were heated on the water bath for less than about thirty minutes, it was difficult to get complete recovery of magnesium. On the other hand if they were heated much longer than this, loss of excess ammonia and evaporation of the alcohol layer occurred. In the best procedure found, an alcohol layer about one centimeter thick was added to the surface of the liquid in the centri- fuge tube just as soon as the precipitate of magnesium hydroxyquinolate started to form. This kept the hydroxyquinolate from creeping up the sides of the tube and prevented the formation of a scum at the surface of the liquid that could not be thrown down by the centrifuge. Two ml. of alcohol were sufficient to form a layer on the surface of the liquid in a 15 ml. centrifuge tube. After heating, the solution should stand for at least one hour before cen- trifuging. A standing period of twenty minutes gave a recovery of only ninety-three per cent for amounts up to 0.02 mg. in determinations on a series of standards ranging between 0.01 and 0.1 mg. magnesium. For amounts greater than 0.02 mg., recovery was complete. Where the precip- itate was allowed to stand over night the results were slightly more con- sistent than for the one-hour period. One of the greatest difficulties encountered in the use of 8-hydroxyquino- line as an analytical reagent is that it precipitates so many metals in both slightly acid or alkaline solutions. The most important interfering metals occurring in plants or in soil extracts are iron, aluminum and calcium and 38 The Kentucky Academy of Science these are easily removed. In some materials, notably corn grain, so much phosphate is present that it may not be removed by ordinary methods and, furthermore, in the precipitation of calcium considerable oxalate is added. The question then arises as to whether the presence of these two ions inter- feres with the determination of magnesium. It has been reported that an excess of oxalate prevents good precipitation and some procedures call for the removal of phosphates. Several experiments were set up to check on this and while it was found that phosphates did not interfere if sufficient time was allowed for precipitation, conflicting data were obtained in the case of oxalate. Early work showed a definite interference when the amount of oxalate was as much as 0.5 ml. of saturated ammonium oxalate per determination under our experimental conditions. More recent work, however, has shown that as much as 1 ml. of the same solution had no ef- fect. We have no entirely satisfactory explanation for this at present but it is believed to be due to the time allowed for precipitation. The data in Table 1 are typical of our more recent results obtained by the method finally adopted. ‘The experiments have been repeated several times and the results for the two methods of estimation of the quinolate were obtained from different experiments. Table 1. The effect of varying amounts of saturated ammonium oxalate on the precipitation of magnesium quinolate. Mg. Vitae: Mg. Magnesium recovered Magnesium. NH#4 Oxalate Colorimetric Volumetric* 415 0 43 41 415 0 41 41 15 0 43 40 41S 0.2 43 41 415 0.2 -42 41 415 0.2 41 42 41S 0.5 42 43 41S 0.5 -42 42 415 0.5 -41 41 415 1.0 -41 42 415 1.0 43 41 415 1.0 41 40 The 8-hydroxyquinoline method finally adopted and described below was used in determining the magnesium content of about twenty samples of corn grain. The samples were analyzed in duplicate and ranged between 0.1 and 0.14 per cent of magnesium. The duplicate samples agreed fairly closely for all constituents determined and the average deviation between the per- centage magnesium among the duplicates amounted to slightly more than three per cent if two samples were discarded that gave a deviation of sixteen * Oxidation using ammonium hexanitrato cerate. Twenty-Eighth and Twenty-Ninth Annual Meetings 39 per cent. Maximum deviation, aside from the two samples listed above which must be rerun, was 7 per cent pons most samples agreed within one to three per cent. Procedure for the Determination of Magnesium in Plant Materials. Place in a 15 ml. conical centrifuge tube (tube #1) an aliquot of the HCl extract of plant ash containing between 0.01 and 0.08 mg. magnesium. Add 1 drop of methyl red indicator and 0.25 ml. glacial acetic acid; bring to pH 5.9 to 6.1 with SN NHiOH. Stir constantly and do not allow the solution to become alkaline. Add 3 drops of a solution of 2 per cent acetic acid that contains 2.5 per cent 8-hydroxyquinoline. Mix thoroly and run © in a few drops of alcohol to break up any scum of iron and aluminum hydroxyquinolate that forms at the surface of the liquid in the tube. Heat for a few minutes after which centrifuge for 10 minutes at 2000 r.p.m. Transfer the supernatant liquid to another tube of the same size (tube #2). Wash the precipitate once with distilled water and transfer the washings to tube #2. Precipitate calcium by adding 1 ml. saturated ammonium oxalate, stirring and allowing to stand several hours. Centrifuge for 10 minutes at 2000 r.p.m. as before and transfer the supernatant liquid con- taining magnesium to a 40 ml. conical centrifuge tube (tube #3) by means of suction, drawing the liquid thru a glass tube constricted at the end and turned up. This tube should reach to the bottom of centrifuge tube #2 and should be attached to tube #3 by means of a two-holed rubber stopper in such a way that the liquid containing magnesium can be drawn over by suction. Wash the calcium precipitate 3 times with a mixture of one part of alcohol and one part of 2 per cent ammonium hydroxide and transfer the washings to tube #3 as described above. Add to the magnesium solution in tube #3 two ml. of a solution con- taining 2 grams of 8-hydroxyquinoline in 100 ml. of ethyl alcohol. Mix well, and then add 2 ml. NHsOH (141) and mix again. Place the tube in a hot water bath at 75° C and when precipitation starts, usually in two or three minutes, add a layer of alcohol 1 cm. thick to the surface of the liquid in the tube. Leave in the bath for about thirty minutes, then place in a rack and set aside for at least one hour (preferably longer). Centrifuge at 1000 r.p.m. for ten minutes, draw off the supernatant liquid with gentle suction, wash the precipitate twice with a solution made up of one part of ethyl alcohol and one part of 5 per cent NH,OH, centrifuging each time and drawing the liquid off as before. After washing is complete, place the tube back in the water bath to evaporate excess alcohol and ammonia. To the dried precipitate of magnesium hydroxyquinolate, add 5, 10 or 20 ml. (depending on the amount of quinolate) of FeCls acetic acid* * The ferric chloride acetic acid reagent contains one-half per cent of FeCls.6H20 in one-half per cent acetic acid. 40 The Kentucky Academy of Science reagent to the tube, stir, and then set aside for two or three hours or until solution is complete and the green color is fully developed. The green so- lution is then diluted one or two to five and its light transmission value is determined by means of a “‘Cenco-Spectrophotolometer” set at 560 millimi- cron wave length with a 1 mm. entrance slit and a 5 millimicron exit slit. From this data the value of the unknown is picked from a standard curve. SUMMARY The use of 8-hydroxyquinoline for the precipitation of magnesium in semi-micro quantities was found to be quite satisfactory. A method was worked out that gave much more reliable results than would be obtained: by precipitating the magnesium as the ammonium phosphate. The hydroxyquinolate could be estimated satisfactorily by either one of two methods: (1) by titrating with ammonium hexanitrato cerate in 2 M perchloric acid; (2) colorimetrically by dissolving the precipitated mag- nesium hydroxyquinolate in a solution containing one-half per cent each of FeCls.6H20 and glacial acetic acid. ‘The colorimetric method was found to obey Beer’s law, to be very sensitive and to give a color that was stable for a period of several days. It was necessary in making up the ferric chloride acetic acid reagent to be sure the concentration of FeCls.6H20 was accurate. Magnesium is precipitated quantitatively from dilute solutions containing ammonium chloride after an excess of 8-hydroxyquinoline and ammonium hydroxide are added. Precipitation can best be carried out in either 15 ml. or 40 ml. conical centrifuge tubes. The solution should be heated in a water bath at 75°C for thirty minutes and should stand in the cold for at least one hour before centrifugating. A layer of alcohol added to the sur- face of the liquid just after precipitation starts, prevents the magnesium hydroxyquinolate from creeping up the sides of the tube and forming a scum at the surface that will not centrifuge down. Iron and aluminum interfere and must be removed, but phosphates and oxalates do not inter- fere if sufficient time is allowed for precipitation. BIBLIOGRAPHY (1) Berg, R., 1927. Determination of magnesium by means of Oxyquin- line. The analyst 52: 431. (2) Hildebrand, J. and Lundell G.; E. F., 1929. Applied inorganic analysis. John Wiley Company. (3) Hoffman, William S., 1937. A colorimetric method for the de- termination of serum magnesium based on hydroxyquinoline precip- itation. Jour. Biol. Chem. 118: 37-45. (4) Lavollay, J., 1935. Bull. Soc. Chem. Biol. 17:432. (5) Nielsen, John P., 1939. Determination of magnesium in biological materials. Ind. and Eng. Chem., Anal.Ed., 11: 649-651. (6) Redmond, J. C. and Bright, H. A., 1931. Determination of mag-. Twenty-Eighth and Twenty-Ninth Annual Meetings 41 nesium in Portland cement and similar materials by the use of 8- hydroxygquinoline. Bur. Standards Jour. Res. 6: 113-120. (7) Wolff, Rene, 1938. A new method for the microdetermination of magnesium in blood serum by the photoelectric colorimeter. Comptes Rendus. Societie de Biologie 127: 1445-6. SLODIES TONS ELUMAN QBARASTLES aN SEN TUCK: I. PRELIMINARY NOTE ON THE OCCURRENCE OF ENDAMOEBA HISHOLYTIEA AND RELAGED SPECIES IN A GROUP OF COLLEGCEHRSRUDENTS: Davip RIcHARD LINCICOME Department of Zoology University of Kentucky Since its discovery by Loesch in 1875 in the feces of a Russian suffering from dysentery, Endamocba histolytica has come to be recognized as an important, pathogenic protozoan parasite of man the world over. In 1913, Walker and Sellards, by feeding cysts of this organism to human volunteers, demonstrated its causal relationship to a type of dysentery in man. Since this time cumulative work by many investigators has shown that this parasite produces damage to the wall of the large bowel and may secondarily invade the liver as well as other organs and tissues of the body. Extended information on the occurrence and distribution of Endamoeba histolytica and other protozoa of man in Kentucky is apparently wanting, and with the exception of an as yet unpublished survey among Berea College students (Personal communication, Headlee, 1942) no extensive investiga- tion of the presence of E. histolytica and other intestinal protozoa of man has been undertaken in this commonwealth. However, isolated cases of E. histolytica infection reported to the U. S. Public Health Service give evidence that the parasite is endemic here. Since December, 1941, an investigation of the occurrence and distribution of intestinal protozoa has been conducted among students at the University of Kentucky at Lexington, and to date some ninety-eight individuals have been studied. The technic employed for the recovery and diagnosis of intestinal protozoa was one which has been developed and routinely recom- mended by Faust and his associates (1938, 1939). It consisted of but a single fecal examination of each individual involving two principal oper- ations: 1) a direct fecal film and 2) a zinc sulphate centrifugal flotation test. The direct film was made first and consisted of the preparation and microscopic study of an iodine-stained, saline suspension of a fleck of feces on a glass slide. 42 The Kentucky Academy of Science The zinc sulphate test was prepared next. It consisted firstly of the preparation of a water suspension of the bulk of the fecal sample. A ten milliliter portion of this suspension was then centrifugalized. The sediment remaining was next mixed with zinc sulphate solution (sp. gr. 1.180) and re-centrifugalized. The material composed of protozoan cysts and light fecal debris floating on the surface film after spinning was removed by a wire loop to a glass slide, stained with iodine and studied microscopically. Seven species of intestinal protozoa were encountered during the study. Endamoeba histolytica was found in 10 per cent of cases while E. coli was found in 23 per cent, Endolimax nana in 24 per cent, Dientamoeba fragilis and Chilomastix mesnili in one per cent, Giardia lamblia in six per cent, and lodamoeba biitschlii in three per cent. Craig (1934) has estimated that from five to ten per cent of human beings in the United States are infected with E. histolytica and, on the whole, this estimate has been substantiated throughout the United States wherever the organism has been studied. Faust (1941) in a study of two hundred post mortem accident cases has demonstrated a 6.44 per cent asymptomatic infection with E. histolytica in the New Orleans population. Recently Faust (1942) in his study on the prevalence of amebiasis in the Western Hemisphere stated that “the actual incidence of amebiasis in the United States is probably considerably higher than that which Craig (1934) and others have estimated. In the population of the United States an average amebiasis incidence as high as 20 per cent may be reached, rather than the accepted figure of 5 to 10 per cent.’ The preliminary data herein presented, although from a relatively small number of cases, indicate that the incidence of E. histolytica infection among University of Kentucky students probably parallels that estimated for the general population. It seems desirable, since Endamoeba histolytica does occur within the population of this State and in view of the fact that this organism may be a dangerous one, to extend the study of the occurrence and distribution of this parasite in man throughout the State. REFERENCES Craig, C. F. 1934. Amebiasis and Amebic Dysentery. Springfield, Illinois. Faust, E. C. 1941. Amebiasis in the New Orleans population as revealed by autopsy examination of accident cases. Amer. J. Trop. Med. 21: 35-48 3 Faust, E. C. 1942. The prevalence of amebiasis in the western hemis- phere. Amer. J. Trop. Med. 22: 93-105 Faust, E. C., D’Antoni, J. S., Odom, V., Miller, M. J., Peres, C., Sawitz, W., Thomen, L. F., Tobie, J. and Walker, J. H. 1938. A critical study of clinical laboratory technics for the diagnosis of protozoan cysts and helminth eggs in feces. Amer. J. Trop. Med. 18: 169-183 Twenty-Eighth and Twenty-Ninth Annual Meetings 43 Faust, E. C., Sawitz, W., Tobie, J., Odom, V., Peres, C. and Linicome, David R. 1939. Comparative efficiency of various technics for the diagnosis of protozoa and helminths in feces. J. of Parasit. 25: 241-262 Headlee, W. H. 1942. Personal communication. THE MEASUREMENT OF RELATIVE INTENSITIES OF SOFT X-RAYS BY THE PHOTOGRAPHIC METHOD CHarLes L. OWENS Department of Physics University of Kentucky Since their discovery by Roentgen in 1895, the ability of x-rays to affect the photographic plate has been known. This was the primary method used in measurements of intensity. The photographic method is not con- sidered as accurate as other methods, but when the others fail, this lesser degree of accuracy can be tolerated. When measuring intensities of soft x-rays, that is, x-rays of long wave length, certain problems arise that are not encountered in the use of hard radiation. When using the longer wave lengths the absorption in air be- comes so great that all measurements have to be made in vacuo. Also these longer wave lengths prohibit the use of any thin films in the apparatus. The chief difficulty in the use of the photographic method is in the calibration of the photographic plate. This may be accomplished in var- ious ways, but the greater the accuracy desired the more difficult the problem. According to the reciprocity law, the exposure may be expressed by the relation, R==1r where I is the intensity and t the time of exposure. More refined measure- ments indicate that the above relation should be written as E=I where m and n have values near unity. The evaluation of these exponents would then serve as one method of calibration, Several studies have been made of the photographic plate for soft x-rays by various authors. The conclusions reached were that the value of m is dependent upon the wave length, type of emulsion used, and conditions of development. Also the variation of the time of exposure seems to be out of the question. The calibration of the plates was accomplished by varying the intensity. In the use of the hot filament type of tube this is accomplished by varying 44 The Kentucky Academy of Science the tube current, all other factors remaining constant. If the intensity is assumed to be directly proportional to the current, then a number of ex- posures may be taken whose intensities bear a known relation to each other. Then by taking an exposure with a portion of the beam absorbed the cur- rent necessary to produce this intensity in vacuo may be determined from the other exposures. The value of I/I, may be determined from these values of current and then from the relation, T=], expat) the absorption coefficient may be calculated. Since all exposures are taken on the same plate any variation in conditions of development does not affect the accuracy of the measurement. In the use of this method there are certain possible sources of error. The change in tube current may change the focusing conditions of the apparatus or the thin layer of tungsten evaporated on the target may alter the in- tensity. However, preliminary measurements showed these effects to be negligible. The apparatus used consisted of an x-ray tube fastened to a grating chamber which was in turn connected to the camera box. Satisfactory con- nections were made by sylphon tubing. The grating was an original ruled grating having a grating space of d=1.664<10 s+ cm. The system was evacuated by a three stage mercury-in-glass pump backed by a rotary oil pump. This combination was capable of producing a pres- sure of 0.04 microns of mercury. A Pirani guage, calibrated against a McLeod guage, was used to determine the pressure. Measurements were made to determine the mass absorption coefficients of aluminum for three different wave lengths. Two measurements were made at a wave length of 8.03 Angstroms and one each at 13.3 and 14.9 Angstroms. From the values of », the mass absorption coefficient, at 8.03 Angstroms, there is some variation; however, this is small compared to the wide variation in values obtained by different authors. This gives reason to believe that the other values are near correct. The results are tabulated below. i in Angstroms in 8.03 333 8.03 345 es: 1239 Lge) 1800 Twenty-Eighth and Twenty-Ninth Annual Meetings 45 ABSTRACTS Relative weights of boiled and unboiled eggs and their yolks before and after drying.*__G. Davis BuckNER, W. M. INsko, JR. AND AMANDA Harms, Agricultural Experiment Station, University of Kentucky.—In connection with certain metabolism experiments with the laying hen, it became necessary to know the comparative weights of certain parts of eggs before and after being boiled for 30 minutes. Relations have been estab- lished between the weights of whole eggs, shells, whites and yolks before and after boiling. Also the alevsicins vie: have been determined between the water and solids in yolks before and after being boiled and after the yolks had been dried to a constant weight in a forced draft electric oven for three hours at 100°C. A study of some of the changes in composition of egg yolk fat during incubation.* G. Davis BucKNER, W. M. INsko, JR. aND AMANDA Harms, Agricultural Experiment Station, University of Kentucky.—Four lots of Rhode Island Red hens were fed rations which varied widely in composition. Eggs from the hens in each lot were obtained and grouped on alternate days for analyses and incubation. ‘The analyses of the fat in the yolk of egg from hens in the four lots before and after incubation are discussed. The effect of formaldehyde fumigation on embryo mortality in the chick.* W. M. INsko, Jr., Dewey G. STEELE aND Cecit M. Hinton, Agricultural Experiment Station, University of Kentucky.—Formaldehyde fumigation of eggs for control of pullorum disease has been well established and its use is increasing each year. Similar treatment for the control of another trouble- some disease, mushy chick disease, is less firmly established because effective control of this second disease Sage oss two or three times stronger treatment than does pullorum. The relationship between strength of fumigant and embryo mortality was studied for treatments ranging from 3 times the concentration commonly recommended for control of pullorum to 7 times this concentration. It is encouraging to report that embryo mortality was not increased appreciably until the concentration of formaldehyde exceeded 4 times the treatment usually employed for the control of pullorum. The most critical period of treatment was in the early stages of incubation, especially the second and third days. The results obtained in the extensive trials referred to above should be of considerable value to those interested in the poultry industry. A genetic analysis of the 1940 Thoroughbred stakes winners.* DEWEY G, STEELE AND Bitty Jackson,f Agricultural Experiment Station, University of Kentucky.—A genetic analysis of the 1940 Thoroughbred stakes winners * Read at 28th Annual Meeting, 1941, Richmond, Ky. { A senior student in the College of Agriculture and Home Economics. 46 The Kentucky Academy of Science has been conducted according to methods devised by Dr. Sewall Wright and Dr. Hugh McPhee. About 200 random two-line pedigrees were traced to the tenth generation and they constitute the raw data for this study. The following things were observed: the average inbreeding, the average inter se relationship, the influence of certain outstanding ancestors, and the yearly span per generation. There was 3.7% inbreeding in ten generation, an amount almost neg- ligible in its genetic effect. The inter se relationship of 7.0% is an estimate of the genetic similarity resulting from common ancestry. The interval per generation on the sire and dam lines was 11.5 and 11.1 years respectively. A more complete account is being published elsewhere.! An outbreak of plantago virus in Burley tobacco.* W. D. VaLLeau AND E. M. JoHNson, Agricultural Experiment Station, University of Kentucky.—The plantago virus is common in Plantago sp. in the vicinity of Lexington, Kentucky. An outbreak of a necrotic virus disease of Burley tobacco was found to be caused by the plantago virus in a variety which responded with the necrotic reaction. In some varieties, the same virus caused a mottle disease. Both types of reaction were found in tobacco plants naturally infected. In one variety of tobacco carrying the N factor from Nicotiana glutinosa, the plantago virus and the tobacco mosaic virus caused a similar reaction but in another N variety the plantago virus produced few minute necrotic spots while the tobacco mosaic virus gave the usual necrotic reaction. The suggestion is made that the plantago virus may have antedated the origin of N. tabacum and that the numerous strains of the tobacco mosaic virus have been derived from the plantago virus by mutation. A study of certain abnormal bones of chickens raised in Kentucky.* G. Davis BucKNER, W. M. Insko, JR. anD AMANDA Harms. Agricultural Experiment Station, University of Kentucky.—Besides perosis, two types of abnormal bone developments were observed in Kentucky for the first time about 1939. Type I was determined to be osteopetrosis or marble bone which had been found in other states. Type II was of the cancerous or tum- erous character and to date has been observed in Kentucky in the ratio of 3 of the osteopetrosis type to one of the other. Approximately 50 instances of these abnormal bones have been reported to the Experiment Station and they seem to be evenly distributed geographically throughout the State. The blood of the live birds affected was normal for the two sexes. Survey seems to indicate an increased number of this condition appearing in Ken- tucky. These abnormal bones are probably not due to calcium limitation in the diet. *Read at 29th Annual Meeting, 1942, Lexington, Ky. 1 Steele. Dewey G. 1942. Are Pedigrees Important? The Blood-Horse, Vol. 8, pp. 574-577. Twenty-Eighth and Twenty-Ninth Annual Meetings 47 The comparison of fats located in different places in the bodies of laying pullet and yearling hen.* G. Davis BUCKNER, W. M. INsko, Jr, AND AMANDA Harms. Agricultural Experiment Station, University of Ken- tucky.—Depot or reserve body fat located subcutaneously, intra-abdominally and extending along the neck, near the trachea and aesophagus, taken from laying pullets and yearling hens when sampled individually or as composite samples have approximately the same iodine number, refractive index, and color for the two ages studied. The reserve fats are chemically different in individual hens and for the different ages. The average iodine number of the uropygeal glands was higher in the yearling hens than in the pullets. How- ever, the average refractive index was a little higher in the pullets. A study of 363 cases of institutional behavior problems in a drug addict population.* E. OHMER Mitton, Jr., United States Public Health Service Hospital, Lexington, Kentucky.—This study was undertaken in an effort to answer the following questions: 1. In what general ways does the adverse behavior group of patients dif- fer from other patients of the institution? 2. In what ways do those patients who repeatedly violate regulations differ from those patients who violate regulations upon only one occasion? The study was divided into two parts. In Part I, a statistical comparison was made between 363 white, male, drug addict patients who were be- havior problems and patients of the general population of the institution. It was found that: the colored patients were the most maladjusted racial group; adverse behavior occurs most frequently near the beginning of the patient’s incarceration; and, both from a psychiatric and custodial point of view, those patients who repeat adverse behavior tend upon their first violation of regulations to commit the most serious offenses. In Part II, 64 white, male, drug addict patients who were reported for adverse behavior were matched with a group of patients who had not been reported for adverse behavior on the basis of age and on the basis of I. Q. as determined by the Wechsler-Bellevue Intelligence Test. The two groups were compared regarding functions measured by the test and no significant differences were found. The measurement of the behavior of teachers and children in the class- room.* JosEPH E. Brewer, Mental Hygiene Clinic, Louisville, Kentucky. This study reports the measurement of certain aspects of the social inter- play between children and teachers in the classroom. A reliable technique was developed for observing and recording in twenty-nine categories the behavior of children in the schoolroom, simultaneously with the recording of the teacher’s dominative and integrative contacts. The teacher’s behavior * Read at 29th Annual Meeting, 1942, Lexington, Ky. 48 The Kentucky Academy of Science was classified according to three degrees of dominative and three degrees of integrative contacts, the definitions being taken from an unpublished study by Anderson. Fifty-nine children in two second grade rooms were observed individually for a total of twenty-four non-consecutive five-minute periods per child. One teacher in comparison with the other used significantly higher fre- quencies of integrative behavior and significantly lower frequencies of dominative behavior. In her room the children were significantly lower in: looking up from work, undetermined child-child social contacts, conforming and non-conforming. Further lower frequencies in child behavior ap- proached significant differences in playing with foreign objects and holding up hands to recite. Significantly higher frequencies were found in volun- tary social contributions and in social contributions in response to others; higher frequencies approached significant differences in answers spon- taneously and in categories relating to problem solving. Some applications of experimental psychology to industry.* §S. SANFORD Dusin anp E. H. Scoriep, Joseph E. Seagram & Sons, Inc., Louisville, Kentucky.—This paper discusses seven applications of experimental psy- chology to the alcoholic beverage industry: 1. The absolute judgment of distilled alcoholic beverages in odor and taste terms. 2. The method of color discrimination. 3. The use of psychophysics in taste and odor problems. 4. The use of rating scales and other statistical techniques. 5. Threshold studies. 6. Psychochemical correlations. The practical study of factors of neutralization, enhancement, and auanpencetitin as they operate in odor and taste combinations. The paper attempts to show that psychology not only has a place in in- dustry, but can actually control fundamental production processes. N e * Read at 29th Annual Meeting, 1942, Lexington, Ky. Twenty-Eighth and Twenty-Ninth Annual Meetings 49 VOLUME 10 INDEX Page ADstractseolumes(0. nos. land’ 2 eee 30 Abstracts, Volume 10, nos. 3 and 4 ‘a SAS 45 Brauer, Alfred, iMicdertion oi development in Bruchid (Coleoptera) embryos by treatment with amino acids ____________ 24 Brewer, Joseph E., The measurement of the behavior of teachers and clildrensinuithe classroom(absctact m2. ssaes mene 47 Buckner, G. Davis, The comparison of fats located in different places in the bodies of laying pullet and yearling hen (abstract) 47 Relative weights of boiled and unboiled eggs and their — yolks before and after drying (abstract) __ 45 A study of certain abnormal bones of chickens raised in Kentucky (abstract) __ panes 8 es 46 A study of some of the changes in composition of egg yolk fat during incubation (abstract) _ 45 Davies, P. A., Origin and development of prickles in Aralia Spinosa ___ 29 De Glazoff, Alexandra, Comparison of assays of domestic and im- ported! aromatic plants and essential oils = 9-—2 22 = 13 Dubin, S. Sanford, Some applications of ee as psychology to indtastiy (@bstract) i aa=ees 48 Errington, B. J., A spectrographic study of the tissues of wobbler lovers (elosieray)) Me 31 Hageman, R. H., Sources of minor Gemeat contaainatioalin’ sandleul- ture experiments with plants (abstracts) _____------ 31 Harms, Amanda, The comparison of fats located in different places in the bodies of laying pullet and yearling hen (abstract) 47 Relative weights of boiled and unboiled eggs and their yolks after drying (SRASEACIC)) ob ech tll lander la CS A study of certain abnormal bones of chickens raised in Rentu clean (abstract) ya ssaaee ee ee ae 46 A study of some of the changes in composition of ege vols tataduring. incubationg (abstract )=sssmun sae 45 Hinton, Cecil M., The effect of formaldehyde fumigation on embryo MOGeAlieys ine the chick (absttact)) |. sss sees 45 Hodge, Edwin S., Spectrochemical analysis (abstract) ______________ 31 A spectrographic study of the tissues of wobbler horses (abstract) _ at ick ellie ee eS oF cee Hodgkiss, W. S., A spectrographic study of the tissues of wobbler thaws (abstract) eae ee ee ee 31 Insko, W. M. Jr., The comparison of fats located in different places in the bodies of laying pullet and yearling hen (abstract) 47 The effect of formaldehyde fumigation on embryo 50 The Kentucky Academy of Science mortality in the chick (abstract) __- 45 Relative weights of boiled and eabouled eggs and their yolks before and after drying (abstract) sas 4S A study of certain abnormal bones of chickens raised in Kentucky (abstract) i.) 2 5 re 46 A study of some of the changes in composition ‘of egg yolk fat during incubation (abstract) _ ae 45 Jackson, Billy, A genetic analysis of the 1940 Thoroughbred stakes win- ners (abstract) _ ss ms = es Kaneo ward | an eee | pepe ee 3 Kolachoy, Paul J., Comparison of assays of domnassie andl imported aromatic plants and essentialmoils = 9252s =s) == ae 13 Lincicome, David R., Studies on human parasites in Kentucky I. Preliminary note on the occurrence of Endamoeba histolytica and related species in a group of college Studentsye eee ee 41 Lovell, Harry B., The bright green bees of the genera Agapostemon, Aromatics, Augochloropsis, and Augochlorella in Kentucky _ see ae tee ena!) McHargue, J. S., Determination of moisture on tobacco (oven method Vise Vacuum desiccator) (alssrvact))) === === eee 32 Sources of minor element contamination in sand culture - experiments with plants (alstract) |. 2 = ee Melnteer, Bs B.) Whe Barrens on Kentuckyee ss ee 7 Milton, E. Ohne Jr., A study of 363 cases of faganeeel sober ice qupbllense | in a drug addict population (abstract) ._____ 47 Minutes of the 29th Annual Meeting, Kentucky Academy of Science, 1942 EE ee a a. JE 1 Newell, E. Bruce, The earliest movements of the Banebid aber and remarks concerning the neuro-motor mechanism develop- Ment of corresponding stages (abstract), 22) a seem 30 Official’ reports, 1941-1942:)" Secretary aes LJ 32s et dreasuren ae eee 4 Owens, Charles, L., The measurement of relative intensities of out x-rays by the photographic method ae 43. Rapp, Karl E., Determination of moisture on tobacco (oven method vs. vacuum desiccator) (abstract) cues eS ei Scofield, E. H., Some applications of experimental psychology to » industry (abstract ) an 2 eI Shacklette, H. T., A review of Rescmeley botanical history (abstract) 30. Steele, Dewey J., The effect of formaldehyde fumigation on embryo moriiey am the chick/ (alssesact)) ——- 9) eee 45. Twenty-Eighth and Twenty-Ninth Annual Meetings A genetic analysis of the 1940 Thoroughbred stakes wannerse (abstract) _ {aaaamuees ean Es Todd, Jack R., The semimicro determination of magnesium as the quinolate using the colorimetric ferric chloride methodie ss = a “aes Weeks, M. E., The semimicro determination of magnesium as the quinolate using the colorimetric ferric chloride method Woodmansee, C. W., Determination of moisture on tobacco (oven method vs. vacuum desiccator method) (abstract) ~--- 51 4S 3) op) S\ TRANSACTIONS OP WHE KENTUCKY ACADEMY OF SCIENCE (Quarterly Series) AG EIEDAT ED > Wileiiiie SAGAC ALS: VOLUME 11—NUMBER 1 JUNE 1943 THIRTIETH ANNUAL MEETING 1943 CONTENTS Report on the Thirtieth Annual Meeting SNES etnies dG Re Secretary's Report (abstract) Sede AME et = Sep BE sel a Some Functions of Mineral Elements in Connection with Enzymatic Action (Presidential address) J. T. Skinner i A MRA ot es Robie Sic WN ay Floral Glands in Ailanthus Altissima pe AND) avatesy2 i): Pt RE ate cg SI ie TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE EDITORIAL STAFF bawerence Baker: 4/12 os. Berea College ____ Psychology and Philosophy EW Cooks 220): Centre: Colleccmes™n a sasuum Bacteriology Hy By; Lovell 22.20 ls University ot veousville, ==. eee Biology Wee: Nichar bart eee eels University of Kentucky 25" a8 Geology Ward GSumpter) 208 25s oa Western State Teachers College om Chemistry atvis: odd asus omit University of Kentucky ___________ Physics John) Kasper: seas / University of Kentucky, Managing Editor Manuscripts. The Transactions must be limited to the proceedings of the annual meetings of the Kentucky Academy of Science and to original manuscripts pertaining to science. Manuscripts are subject to the ap-— proval of the Editorial Staff and may be submitted to the Editor of Be subject covered or to the Managing Editor. ExTra-Cost Features. The extra cost of special features such as cuts, graphs, tables, etc., above the text-run price per page must be borne by the contributor. The Editorial Staff will advise contributors concerning the extra cost of features upon receipt of manuscript. Illustrations to be in- cluded in an article should accompany the manuscript if possible, or, if sent in separate package should be properly labeled as to the article in which they are to occur. Proor. Galley proof will be sent for approval of contributors. The proof should be returned promptly to the Berea College Press, Berea, Ken- tucky. REPRINTS. Reprints are furnished at publisher’s prices by negotiating directly with Berea College Press. Price quotations on reprints are sub- mitted with the proof. Orders for reprints should accompany the proof to the Berea College Press, Berea, Kentucky. SUBSCRIPTION Rates. The Transactions is sent without additional ex- pense to all members of the Kentucky Academy of Science who are not in arrears for annual dues. The annual subscription rate for non-members is $2.00 in the United States and Canada, $2.50 in foreign countries; ‘single numbers 75 cents. One volume of four numbers appears each Academy fiscal year. BusINESs CORRESPONDENCE. Remittances and correspondence concern- ing subscriptions, extra costs, and other financial matters except reprints should be addressed to John Kuiper, Sa of Kentucky, Lexington, Kentucky. \ H\'ke aM 70st OTN _ —! beh, VAL. Wi Uo! - — Oe cr a a a by ae Se ae } | LO wr? Dope QAM \ ' my Oo | 4 , 0S er et ae Thirtieth Annual Meeting 1 THIRTEENTH ANNUAL MEETING ~ Louisville, Kentucky April 23-24, 1943 First General Session The first general session of the Kentucky Academy of Science was called to order by President J. T. Skinner on Friday, April 23, at 2:20 p.m., in Allen Court Room, Law Building, University of Louisville. The following reports were heard: Reading of minutes by the Secretary. Secretary’s report, Alfred Brauer. Treasurer’s report, Wm. J. Moore. Councilor’s report on Junior Academy activities, Anna A. Schnieb. Representative on Council of the A.A.A.S., Austin R. Middleton. Managing Editor’s report of the Academy Transactions, John Kuiper. No RWND In this report the following possible methods were suggested for placing the publication on a sound financial footing. 1) Increasing the membership 2) Obtaining advertising for the Transactions of the Academy 3) Soliciting the support of the State’s educational institutions. A motion was made from the floor and seconded, that the President appoint a committee to investigate the possibilities of soliciting the support of educational and industrial institutions, and that the committee submit a report at the second business session. "[Thé motion was carried. President Skinner then appointed the following committee: Morris Scherago, Chairman; Paul Kolachov; John Kuiper. 7. Report of the Chairman of the Membership Committee, Ruth N. - Fonaroff. The names of 30 candidates for membership were read at this time, and these were subsequently elected. Later in the day, the Executive Com- mittee received the names of others, who were elected by the Committee. This entire list is given elsewhere. 8. A report of the Special Committee on a constitutional amendment was read. The report proposed an amendment relative to the appointment of a board of editors for the Transactions. The report was tabled. A request for affiliation in the Kentucky fade of Science was received from the Kentucky Society of Natural History. \t was read by Mr. R. E. Stouder, who is also a member of the Academy. Action on this request was deferred to the Second Business Session to be considered under new business. 2 The Kentucky Academy of Science The meeting adjourned for the address of the president Some Functions of Mineral Elements in Connection with Enzymatic Action Dr. J. T. Skinner Second General Session Allen Court Room, U. of Louisville, Saturday, April 24, 8:00 a.m. President Skinner called the meeting to order and asked that the minutes of the preceding meeting be read. These were read and approved. The minutes of the Executive Committee meeting of April 23, 1944, were then read. These contained and provided for the following recom- mendations: 1. Placement of the A.A.A.S. fund for aid in research as follows: To Dr. D. R. Lincicome, $50.00 for purchase of materials required in his study of certain protozoan diseases in Kentucky. To Dr. W. R. Allen, $32.50, for the purchase of an accessory flat- field lens, for the preparation of plates in his study of Kentucky fishes. 2. Acceptance into affiliation with the Academy of the Kentucky Society of Natural History. 3. That Dr. Schnieb again be allowed a sum not to exceed $30.00 for expenses incurred by the Kentucky Junior Academy of Science. 4. That the names of the members of the Jos. E. Seagrams & Sons, Inc., as proposed by Mr. Herman F. Willkie, be accepted into membership of the Academy. The motion was made and seconded that this report be accepted. After discussion of each of the several items the minutes, including the recom- mendations, were voted upon and adopted. ; By a unanimous vote the question of an amendment to ARTICLE X of the constitution, as proposed by the Special Committee was raised from the table. The proposed amendment is as follows: Section 1. The President shall appoint an editorial board composed of a managing editor and a staff of assistant editors. One assistant editor shall be appointed from each of the major divisions of the Academy. Section 2. The editorial board shall have the power to select the papers to be published and to reject any which it does not deem suitable for publication in the Transactions. Section 3. That the editorial board be authorized to return papers to their authors when necessary requesting reduction in length or other modifications. Section 4. That papers accepted for publication be limited to five pages of print, except the president’s address, unless the author pays in ad- vance for extra pages. Also that all special features such as figures and expensive tables be financed by the author. A short but vigorous discussion of the proposed amendment followed. A motion was then made that sections 2,3, and 4 be eliminated from ‘Thirtieth Annual Meeting 3 the proposed amendment, and that section 1 be reworded by the Executive Committee and made to conform with the original ARTICLE X of the Constitution. The motion was seconded and passed. A report by the Committee on Sustaining Memberships was called for. The following report was read by the chairman: “Sustaining membership: Sustaining membership may be extended to any educational or industrial institution in Kentucky interested in the pro- motion and advancement of science, and in sympathy with the objectives of the Kentucky Academy of Science. An institution holding a sustaining membership may designate one of its members to represent it as a voting member and shall otherwise enjoy all of the privileges of membership. The annual dues for a sustaining member shall be $10.00. : It shall be the duty of the executive committee to prepare a list of institutions eligible to sustaining membership. The President shall extend the invitation to the institutions listed and shall include a statement of mutual advantages of affiliation.” It recommended that the Executive Gammmibeee take the necessary steps for establishing such membership, and made the motion that this be done. The motion was seconded and after discussion, was passed. A report of the Nominating Committee was called for. This proposed the following slate of officers for the year 1943-1944: President, L. A. Brown, Transylvania College Vice-Pres., Paul J. Kolachov, Seagrams & Sons, Louisville Secretary, Alfred Brauer, University of Kentucky - Treasurer, William J. Moore, Eastern Ky. State Teachers College Rep. on Council of the A.A.A.S., Austin R. Middleton, University of Louisville For Directors to serve till 1947, G. B. Pennebaker, Morehead State Teachers College L. Y. Lancaster, Western Kentucky State Teachers College L. S. O'Bannon, University of Kentucky, was nominated from the floor. Messrs. Pennebaker and Lancaster were elected. A report of the Resolutions Committee was called for and read by the Chairman, M. L. Billings. Other members of the committee are, Lucien Beckner, W. R. Allen. The report was adopted. The resolutions are: 1. A resolution of thanks and appreciation to the University of Louis- ville for its hospitality and the use of its buildings for the meetings. 2. A resolution of sympathy to the University of Louisville for the loss by sudden death of its President, Raymond A. Kent. 3. A resolution of sympathy to the University of Kentucky at the loss by death of Dr. James Burt Miner, Head of the Department of Psy- chology, and an active member of the Kentucky Academy of Science. 4 The Kentucky Academy of Science 4. A resolution expressing pride in the growth and expansion of scientific interest in Jefferson County, expressed by the organization of the many scientific clubs. 5. A resolution expressing satisfaction in the number of clubs and organizations throughout Kentucky whose programs are directed toward the conservation of natural resources. 6. A resolution expressing pride in the contribution which many of the members of the Academy are making in the war effort, and especially in the number which the Academy has contributed to the armed forces. ABSTRACT of The Secretary’s Report to the Kentucky Academy of Science Membership: Total membership _____________ 4) 307, Actual sustaining members ___________ 262 Llonoranys 92 22) - Neem 72 Emeritus Me 3 Corresponding a aa ee Life iu cs 3 National i Es 88 Absent for period cf the war =e Those now serving in the uniformed services and their branches of service are as follows: Charles E. Bortner, Army; Curtis H. Bottom, Ensign, Navy; Lucille Catlett, Lieutenant, WAVES; Minor E. Clark, Army; Robert L. Driver, 1st. Lt. Army; Harold T. Glover, Capt., Army; T. M. Hahn, Major, Army; D. P. Hall, Army; Lawrence Henson, Army; J. E. Hernandez, Capt., Army; Milton B. Jensen, Capt., Army; J. L. Keffer, Army; Durbin C. Kemper, Lt., Army; J. W. Lancaster, Capt., Army; Forrest Mercer, Coast Guard; Wayland Rhoads, Major, Army; Dean W. Rumbold, Army; H. T. Shacklette, Lt., Army; Wm. L. Smith, Navy; Jack R. Todd, Army; S. J. Lowry, Army. Research Grants: @irried wover trom: 947 eee eeeeees $42.00 Amount for 1943 _ 2 8550 Awailable for 1943, total 220 $80.00 Projects completed on former grants, all conditions of grants having been met: . B. B. McInteer and H. T. Shacklette in The Bryologist,45: 125-134. Ward C. Sumpter in Jour. Am. Chem. Soc.,64:1736. Thirtieth Annual Meeting ) Death of Members: Arthur F. Fehn, Centre College (Former member) Dr. James Burt Miner, University of Kentucky King Award The King Award of $50.00 for the most meritorious paper presented at the 1942 annual meeting was made to M. E. Weeks and Jack Todd for their paper on “The semimicro determination of magnesium as the quinolate using the colorimetric ferric chloride method.” | SOME FUNCTIONS OF MINERAL ELEMENTS IN CONNECTION WITH ENZYMATIC ACTION* J. T. SKINNER Agricultural Experiment Station University of Kentucky The term mineral element includes any of the approximately ninety simple substances, exclusive of carbon and oxygen, which remain wholly or in part in the ash when plant or animal material is burned. Except when proper precautions are taken to insure their retention in the ash a considerable proportion of some of the elements, e.g. sulfur and chlorine, may be volatilized at the temperatures imposed during oxidation. Mineral elements found most abundantly in biological materials are calcium, mag- nesium, potassium, sodium, iron, phosphorus, sulfur and chlorine. Occurring in much lower concentrations, although apparently also universally dis- tributed, are aluminum, copper, cobalt, zinc, manganese, arsenic, boron, _ iodine, fluorine and probably several others yet to be determined. Whereas the term mineral element suggests occurrence of an element in compounds closely akin to minerals of the earth’s crust, such elements occur in living organisms frequently as constituent parts of relatively com- plex structures. Phosphorus, for example, in addition to occurring in bones and teeth of animals in the form of a mineral compound similar to rock phosphate is found in biological materials combined with carbohy- drates, lipides, proteins, nucleosides, vitamins, nitrogen bases and alcohols. Having determined that a given element is of universal occurrence in plant or animal tissue, or perhaps in both, the next task of the biochemist is to ascertain whether its presence is essential to the welfare of any organ- ism. Fortunately response in rate of growth of a young plant or animal is a fair index of the essential nature of a chemical element. Perhaps this criterion would be infallible if 100 per cent of the element could be elim- * Presidential address, Thirtieth Annual Meeting, April 23-24, 1943. 6 The Kentucky Academy of Science inated from the nutrient medium of the plant or the diet of the animal, but the last traces are indeed difficult to remove. Assuming, however, that a given element is found to be essential for normal growth there re- mains yet to be accomplished the more difficult task of determining the specific role or roles played by that element in the organism. Some of the more familiar roles of mineral elements are (1) formation of bones and teeth, (2) maintenance of osmotic pressure, (3) establish- ment of a proper balance between acidic and basic elements, (4) buffer action, and (5) formation of chlorophyll of plants and hemoglobin of animals. In addition to performing these five well known functions mineral elements participate in the formation of structural units of the soft tissues, notably sulfur of proteins; they are responsible for irritability of muscle and nerve; they sometimes occur in hormones, as is true of iodine in thyroxine; and, finally, they are involved in digestion and absorption of the organic constituents of foodstuffs and in those processes by which such — constituents are metabolized with consequent release of energy to the organ- ism. In discussing the last named functions, which may be restated thus: “Some Functions of Mineral Elements in Connection with Enzymatic Action,” it will be necessary to touch upon the chemistry of several classes of complex organic compounds such as proteins, nucleotides and vitamins. One of the most useful processes by which chemists separate pure substances from mixtures is that of crystallization. During the past two decades rapid strides have been made in the isolation in crystalline form of enzymes from plant and animal materials. Included among the enzymes isolated as crystals relatively free from contaminating materials are urease, pepsin, pepsinogen, trypsin, trypsinogen, chymotrypsin, chymotrypsinogen, papain, muscle phosphorylase, ascorbic acid oxidase, catalase, peroxidase, various flavoproteins, carboxypeptidase, amylase, ficin, enolase, phospho- glyceraldehyde dehydrogenase, ribonuclease, alcohol dehydrogenase, and lac- tic acid dehydrogenase. All enzymes thus far obtained in crystalline state have proven to be proteins and in view of the general behavior of many other highly potent enzyme preparations it appears justifiable to assume that most, perhaps all, enzymes are protein complexes. If they qualify as simple proteins by yielding only amino acids when hydrolyzed, sulfur is the only mineral element found therein. They may, however, belong to the class of conju- gated proteins, which result from union of simple proteins with suitable prosthetic groups. In these conjugated proteins occurrence of mineral elements is limited to such prosthetic groups, or annexes if you please. Even so, such elements constitute essential parts of the molecular structures of these enzymes. At least three enzymes, catalase, peroxidase, and cytochrome oxidase, Thirtieth Annual Meeting 7 contain iron in their prosthetic groups as does the familiar protein hemo- globin. In fact, determination of the structure of hematin obtained from this readily available blood protein contributed greatly to the solution of the chemical nature of these iron-containing enzymes. Iron in such com- plexes may undergo valence change from two to three and vice versa as it does in its simple inorganic compounds. The function of catalase—don’t forget the iron in its molecular structure—appears to be chiefly that of ‘catalyzing the breakdown of hydro- gen peroxide to water and oxygen. A typical example of this catalysis, one with which all are familiar, is the behavior of hydrogen peroxide when applied to a wound. Bubbling is caused by the escape of oxygen liberated through the enzymatic action of body catalase and is not indicative of in- fection. The universal occurrence of catalase suggests that nature has provided tissues with this agent as a protective measure against accumula- tion of hydrogen peroxide in toxic concentrations. Rapidity of enzymatic action is well exemplified by catalase, one molecule of which decomposes 44,000 molecules of hydrogen peroxide per second at O°. Expressed in terms of iron, which constitutes about 0.1 per cent of the weight of the enzyme, 760 cc. of oxygen gas can be liberated per second at O° by the action of one milligram of catalase iron upon hydrogen peroxide. Peroxidase, although structurally related to catalase, differs from it in occurrence and action. Apparently occurring solely in plants, it func- tions in transfer of oxygen from hydrogen peroxide or some organic per- oxide to an oxidizable substance. Examples of familiar biological compounds susceptible to peroxidase action are tryptophane, histidine, tyrosine, adrena- line, and ascorbic acid. Cytochrome oxidase, known also as indophenol oxidase, catalyzes oxidation of the cytochromes by molecular oxygen. Reference will be made to this reaction in a later section of the discussion. In order to facilitate removal of carbon dioxide from the body, it is highly essential that the gas be taken into solution immediately following its liberation in the tissues. Equally important in the lungs is rapid reformation of gaseous carbon dioxide. Nature’s solution of the problem is an enzyme which hastens establishment of an equilibrium between water and gaseous carbon dioxide on the one hand and carbonic acid on the other. In constructing a molecule suited to the task she synthesized a conjugated protein containing zinc. This enzyme is known as carbonic anhydrase. Zinc has also been reported to be an essential part of intestinal phosphatase, an enzyme catalyzing the hydrolysis of certain organic phosporus com- pounds. Rather widely distributed in the plant kingdom are several copper- containing enzymes belonging to the family of oxidases. During the course of their catalytic activity the copper in these enzymes changes valence from cupric to cuprous state and vice versa. These plant oxidases, of which 8 The Kentucky Academy of Science monophenol oxidase (or tyrosinase), polyphenol oxidase, and laccase are best known, catalyze the direct union of molecular oxygen with certain benzenoid compounds. The familiar blackening of potatoes when a cut surface is exposed to air is the result of such a reaction. That copper functions in enzyme systems of all animals is highly probable. Tissues of arthropods and certain molluscs are known to contain enzymes similar to polyphenol oxidase and laccase. Utilization of iron by vertebrates in formation of hemoglobin, cytochromes, and cytochrome oxidase is dependent upon an adequate intake of copper. Although direct evidence is lacking, it may well be that a copper-containing enzyme plays a role in the synthesis of these important compounds of iron. The metallic element magnesium is said to be an essential part of the enolase molecule which is involved in carbohydrate breakdown. Why are mineral elements contained in enzymes? What functions do they perform there? In some enzymes containing copper and iron the ability to influence reaction rates is dependent upon valence change of these min- eral constituents but valence change cannot be the only significant con- tribution of metals to the activity of enzymes. Some metallic elements occurring in enzymes do not exist in two valence states and, furthermore, iron of peroxidase does not undergo valence change during catalysis by this enzyme. Regardless of dearth of information on the subject, in many and perhaps in all enzymes belonging to metalloproteins the metals apparently perform important roles in enzymatic action and are not incorporated in the molecules simply as so much structural material. The tendency of an enzyme to catalyze a reaction which no other enzyme will affect is termed specificity. In spite of the fact that enzymes belonging to a given family may be quite similar in chemical structure, the individual members exhibit this characteristic to a marked degree. Such specialization among biocatalysts obviously must be due to peculiarities in structural design. In some instances specificity has been traced to -SH (thiol) groups contained in the enzyme molecules—Note the sulfur. Apparently these reactive groups. participate in formation of labile com- plexes between enzymes and substrates, but the exact mode of labilization is unknown. Needless to say this constitutes relatively an important phase of enzyme research. The activity of an enzyme often is markedly affected by the presence of one or more inorganic ions. Any such ion which increases the activity of a given enzyme is termed its activater. It would appear to be an easy task to summarize the role of mineral elements in this capacity but search through the literature reveals a bewildering array of conflicting reports from many laboratories. So far as ability to “hese nits reports is concerned, it is unfortunate that a given ion when present in minute amounts may activate an enzyme whereas at higher concentrations it may exert no effect or even inhibit enzyme action. Complicating the situation still further Thirtieth Annual Meeting 9 is the fact that more than one kind of ion may serve as an activator of a particular enzyme. In other words, there is less specificity of activator for enzyme than there is of enzyme for substrate. Finally, in some instances there is a question whether an element merely activates a particular enzyme or indeed forms an essential part of the molecular structure in which it is held so loosely that purification procedures usually result in its removal. Aluminum, calcium, chlorine, iron, magnesium, manganese and zinc have been reported most often to be activators of enzyme systems. One may well wonder by what mechanism inorganic ions impart activity to enzymes. Hellerman has explained activation on the basis of metal coordination. That is to say, with the aid of certain functional groups in enzyme and substrate the metallic ion serves as a connecting link between the two compounds. It is conceivable that fixation of chemical bonds thus brought about in the substrate would weaken the structure at some point and thereby facilitate chemical change. One of the most important steps in unraveling the mystery of metabol- ism of carbohydrates was the discovery that phosphorus as phosphate is essential to conversion of glucose into alcohol through the action of enzymes of yeast juice. Subsequently it was shown that in the animal body glucose must be converted first into hexose phosphate which then may be taken stepwise through a number of intermediary compounds, many of which are esters of phosphoric acid, until eventually carbon dioxide and water result. It is claimed that during oxidation of a molecule of glucose by the body removal of at least ten of the twelve hydrogen atoms is directly dependent upon phosphorylation reactions, i.e. formation of phosphate esters. Nature’s extensive use of phosphate esters to form labile derivatives of many metabolites is due probably to the ease with which linkage between enzyme and substrate can thereby be effected. Because of the polybasic character of phosphoric acid its esters contain acidic hydrogen and hence may combine readily with proteins, including the specific enzymes which catalyze oxidation of these esters. The simplest type of biological oxidation in which an enzyme can participate consists of simple addition of oxygen to the metabolite or re- moval of hydrogen therefrom through union with molecular oxygen. So far as food materials are concerned this simple oxidation apparently is seldom realized. Instead certain complex heat-stable compounds serve as carriers of hydrogen released by the metabolite, passing it along under the influence of appropriate enzymes, known as dehydrogenases, from one carrier to another until eventually it combines with oxygen. Such complex heat-stable compounds which must be present for enzymes to exert their catalytic effect are termed coenzymes. Invariably they contain one or more mineral elements, but they must not be confused with activators which are merely ions of mineral elements. Although the term carrier perhaps conveys the meaning quite ade- 10 The Kentucky Academy of Science quately, a homely illustration of their role may not. be amiss. Let us assume that the author finds himself in great need of some ready cash which he seeks to raise by disposal of his library consisting of Chemical Abstracts collected over a period of years, some fiction which his family has read, and quite an array of old textbooks representing an accumulation of those studied and taught plus many free examination copies, now out of date, received from a number of publishing houses. One of my associates in the Academy learns of my plight and buys Chemical Abstracts. He in turn, because of his affiliation with the American Chemical Society, sells the entire lot to that society. Here they are passed on to various individuals who have requested back numbers to complete their files. The appearance of my library has been changed and it is no longer worth as much as at the be- ginning, but it can yet be made to yield some cash. As a friendly gesture a neighbor of mine buys the fiction, which he donates to the army when informed by his soldier son of the need of service men for reading material. Only my old textbooks remain as a source of cash, energy if you please. One of my former colleagues becomes sufficiently concerned over my dire need to loosen his purse strings a bit and take these books off my hands. Through his church affiliation he learns of a settlement school which is in need of reference books of all kinds. Consequently he ships the entire lot to this school. It is doubtful if I unaided could have made a single sale to any individual who eventually acquired a part of my library. It was through the intermediary action of various individuals (the co-enzymes or carriers if you please) prompted by certain influences (enzymes) that the money (energy) was made available little by little (stepwise dehydrogena- tion) to me (the organism). Even though the picture as a whole may be somewhat inaccurate, it tends to illustrate the contribution which coen- zymes, and hence certain mineral elements, make to enzymatic action. At least six compounds deserve mention in any discussion of coenzymes. They are diphosphopyridine nucleotide, triphosphopyridine nucleotide, flavin nucleotides, the cytochromes, glutathione, and cocarboxylase. Diphosphopyridine nucleotide or simply DPN is known also as cozymase and coenzyme J. A molecule of adenine, two molecules of the pentose sugar ribose, one molecule of the amide of nicotinic acid (a member of the vitamin-B complex), and two molecules of phosphoric acid are combined to form this nucleotide. It functions as a hydrogen carrier by virtue of the ease with which the pyridine ring of the nicotinamide fraction acquires two hydrogen atoms and transfers them to some other compound. Since no mineral element is located in the ring where this hydrogén transfer occurs, one could assume that the oxidation-reduction change is entirely independent of the mineral component of the molecule. A study of related synthetic compounds reveals, however, that coenzyme activity is lost when any part of the molecule, including phosphoric acid, is omitted from the structure. Among dehydrogenases for which this nucleotide serves as coenzyme are Thirtieth Annual Meeting 11 those acting upon lactic, malic, and B-hydroxybutric acids, triosephosphate, glycerophosphate, and glyceraldehyde. Triphosphopyridine nucleotide, abbreviated TPN and also termed coenzyme II, differs from the preceding nucleotide only in that three rather than two molecules of phosphoric acid are found therein. In view of the almost identical structures of the two it is strange indeed that with but two exceptions recorded to date neither of these coenzymes can replace the other as hydrogen carrier. TPN acts as coenzyme for hexose monophosphate, citric acid, and phosphohexonic acid enzymes. In at least two coenzymes both phosphoric acid and the vitamin riboflavin are essential constituents. They are riboflavin phosphate and flavinadenine dinucleotide. One of the rings of the riboflavin molecule is particularly susceptible to reduction and thus it confers upon its phosphate derivatives ability to act as hydrogen carriers. Coenzymes containing riboflavin exhibit in nature a marked tendency to unite with various pro- teins to form relatively stable complexes known as flavoproteins. One of the most important functions of such flavoproteins appears to be that of transferring hydrogen from reduced pyridine nucleotides, i.e. DPN and TPN. to the cytochromes. Occurring in all cells are three iron-porphyrin compounds known as cytochromes, the importance of which is suggested by the fact that they occur in highest concentrations in those tissues which are most active. Although discovered in 1886 their physiological significance remained un- known until 1925. They too are characterized by their ability to undergo reversible oxidation and reduction. Whereas the three coenzymes previous- ly described aid in oxidation merely by transport of hydrogen, the best known cytochrome, viz c, effects an actual union between this transported hydrogen and molecular oxygen. In the tissues this reaction of reduced cytochrome c with molecular oxygen is dependent upon the catalytic action of cytochreme oxidase, an iron-containing enzyme mentioned previously. In reduced form glutathione is a tripeptide of three amino acids— glycine, cysteine, and glutamic acid. The occurrence of a sulfur atom in the cysteine fraction enables glutathione to undergo oxidation and reduction with ease. In spite of its general occurrence and extensive investigations of the chemical behavior of this compound, little is known regarding its specific role in oxidation of body metabolites. Parenthetically I may say that glutathione functions as a protective agent for vitamin C by main- taining it in reduced form and thereby preventing its irreversible oxidation to compounds exhibiting no vitamin activity. Finally, in this group of coenzymes belongs a complex of another B vitamin, viz diphosphothiamin or thiamin pyrophosphate. This coenzyme contains two mineral elements, phosphorus and sulfur. Prior to the eluci- dation of its structure the compound was named cocarboxylase to indicate its relationship to the enzyme of yeast which splits out carbon dioxide from 12 The Kentucky Academy of Science pyruvic acid. In the animal body its chief function appears to be that of a hydrogen carrier in the oxidative breakdown of alpha-ketonic acids of which pyruvic acid is perhaps the most familiar member. In summarizing we note that (1) mineral elements function as struc- tural units in enzyme molecules; (2) specificity in some instances can be attributed to a chemical group containing sulfur; (3) mineral elements activate many enzymes; (4) phosphorus derivatives of metabolites serve as labile intermediates susceptible to enzymatic action; and, finally, min- eral elements form parts of the molecular structures of coenzymes or hydro- gen carriers which are essential to biological oxidations. Since life is im- possible without the continued performance of the enzyme systems of the organism, any discussion of functions of mineral elements in plants or animals is far from complete if it fails to include the contributions which these elements make to enzymatic action. FLORAL GLANDS IN AILANTHUS ALTISSIMA P. A. Davies University of Louisville Following its importation into North America, Ailanthus altissima Swingle (tree-of-heaven) had a favorable reception. It has many desirable characteristics for city planting: it is a medium size tree with a heavy subtropical foliage, it will withstand smoke and dust better than most trees, it is almost free from fungus and insect attacks, and will grow on almost all types of soil. However, it has several unfavorable character- istics, chief among these is a disagreeable odor. Although the odor arises from both floral and extra floral glandular structures, it is the odor from the floral glands which is most prominent. Saadies sini ineve [been made on the floral glands of Ailanthus altissima have come from the taxonomists rather than the morphologists or physiologists. Green (1939) states that when the staminate flowers open, we realize the cause of the unpopularity of the trees, for the odor is overpowering and far reaching, but that the pistillate or seedbearing flowers do not have an unpleasant odor. Britton and Brown (1913), Jepson (1923), Rehder (1927), and Bailey (1935) mention the disagreeable odor arising from the staminate flowers but fail to mention any odor arising from the pistillate flowers. The purpose of this study is to present data on the position, structure, and secretion of glands in both staminate and pistillate flowers and to cor- rect certain misconceptions prevalent in the literature. Glandular studies were made in two ways: (1) by direct observations on freshly gathered flowers, and (2) through the preparation of micro- Thirtieth Annual Meeting 13 sections. For direct observations of glands on the disc, the pedicel was embedded in modeling clay to hold the flower in an upright position. Any structures as petals or stamens which obstructed a clear view of the disc were removed. For glandular structures below the disc the sepals and lower part of the receptacle were removed and the remainder inverted and pressed firmly into modeling clay. Micro-sections were prepared by fixing in formalin-acetic-alcohol, as given by Chamberlain (1930), dehydrated by the Zirkle (1930) method, embedded in paraffin, sectioned, stained with crystal violet, and mounted in balsam. The staminate and pistillate flowers are borne in terminal upright panicles on different trees. The staminate panicles are larger than the pistillate panicles and contain more flowers. The average number of potential flowers present in each of 50 pistillate and 50 staminate panicles for three consecutive years was: the first year, 199 pistillate and 1398 staminate flowers; the second year, 126 pistillate and 1161 staminate flowers; and the third year, 112 pistillate and 907 staminate flowers. The ratio was 1:7, 1:9, and 1:8. This indicated that there was on the average eight times as many staminate flowers as pistillate flowers,in a panicle. The flowers in a panicle do not ail open at the same time. Over a three year period, the average time from the beginning to the end of flower- ing in staminate panicle was 27 days and in pistillate panicle was 12 days. On the average, the first staminate flower opened three days before the first pistillate flower and continued to open 12 days after the last pistillate flower. After opening, the staminate flowers remained for an average of only 2.9 days and then fell from the panicle. By the time the last staminate flower had fallen, the fruits averaged 2.6 cm. in length. The flowers are small and usually regular with 5 spreading greenish petals. The spread of the petals in staminate flowers averaged for 500 measurements 7.1 mm. and for pistillate 6.9 mm. The normal staminate flowers have 10 fertile stamens inserted in two cycles at the base of a 10 lobed disc and a rudimentary ovary, while the pistillate flowers have a 5 cleft ovary and 10 non-functional stamens. Two different secreting areas are present in pistillate flowers, stigmatic and basal (fig. 1), while only the basal is present in staminate flowers. The stigmatic gland exhibits nothing unusual and serves the same function as that observed from other species. The basal glands appear in three cycles about the base of the flowers: on the disc, between the petals, and between the sepals. The basic number of glands is 15, 5 in each cycle, although a variation from this number may be present. Fig. 2 shows an apical view of the disc glands in a staminate flower. The 5 disc glands are closely packed and form a ring about the rudimentary ovary. They extend from the rudimentary ovary to and partly surround the filament of the stamens, and press firmly against the interpetal glands. The five disc glands are frequently broken up into many glands. The interpetal glands ® FON OAC, Fig.3 Fig. 1. Median longitudinal section thrcugh a pistillate flower showing the position of the stigmatic and basal glands. Legend: stig, stigma;, pet, petal; fil, filament; d.g., dise gland; i.p.g., interpetal gland; sep, sepal. Fig. 2. An apical view of a staminate flower showing the position of the glands. Legend: i.p.g., interpetal glands; d.g., disc glands; pet, petal; fil, filaments. Fig. 3. A basal view above the sepals of a staminate flower showing the position of the glands. Legend: pet, petal; i.p.g., interpetal glands; rec, receptacle; i.s.g., intersepal glands. Fig. 4. An apical view of a pistillate flower after the ovary had been removed showing the position of the glands. Legend: i.p.g., interpetal glands; d.g., disc gland; pet, petal; fil, filaments. 3 Fig. 5. A basal view above the sepals of a pistillate flower showing the position of the glands. Legend: pet, petals; i.s.g., intersepal gland; i.p.g., interpetal glands; rec, receptacle. Fig. 6. A section through an interpetal gland. Thirtieth Annual Meeting “185 lie opposite the disc glands, extend from below, and partly surround the filaments. Fig. 3 is an inverted staminate flower showing the position of the interpetal and intersepal glands. The interpetal glands lie between the petals, almost surrounding them from below and extending upward to the margins of the disc glands. The intersepal glands are small, wedge shaped and lie between the lower margins of the interpetal glands. A disc view of the glands in a pistillate flower is shown in fig. 4. The disc gland is not divided as in the staminate flower but is a single large gland surrounding the ovary. It extends outward beyond the inner cycle of stamens and presses firmly against the interpetal glands. The position of the interpetal and intersepal glands below the petals in a pistillate flower is shown in fig. 5. The interpetal glands are of the same number and occupy a similar position as those presented in the staminate flower. The intersepal glands varied from 1 to 5 and occupied a similar position between the lower margins of + the interpetal glands as was found in the staminate flowers. Micro-sections of the basal glands of both staminate and pistillate flowers show similar structures. A section through an interpetal gland is shown in fig. 6. The cells are closely packed. They vary in size and shape, and have large nuclei. The glands are devoid of any conducting elements, the secretion is exuded through the glandular mass and exposed upon the surface. i Although there are usually more glands in the staminate flowers, the union of glands and the greater swollen surface of the glands in pistillate flowers yield as much if not more glandular surface than is present in the staminate flowers. . Tests for the pungency of glandular secretion were made on two consecutive years. In the first year’s test 200 staminate and 200 pistillate flowers were placed in each of five 150 cc. beakers. The beakers were paired. Six persons were asked to select the one in each pair which had the most pungent odor. Of the 30 possible selections, 27 selected either the pistillate or no selection could be made. In only three cases was the staminate selected. In duplicating the tests the second year, 5 persons who had not been used the previous year were employed. Of the 25 possible selections, 22 selected the pistillate flowers or were unable to decide, and only three selected the staminate. The tests indicate that the pungent odor arising from the pistillate flowers is as great or greater than that from the staminate flowers. These results are not in agreement with Green (1939) that the pistillate flowers do not have an unpleasant odor. Summary 1. Staminate and pistillate flowers are borne in separate panicles on different trees. Approximately 8 times as many staminate as pistillate flowers are borne ih a panicle. 2. The average time cf flowering for all staminate flowers was 27 days and for all pistillate flowers was 12 days. The staminate flowers began 16 The Kentucky Academy of Science to open 3 days before the pistillate flowers and continued to open 12 days after the last pistillate flower. 3. Glands are present in three cycles about the base in both staminate and pistillate flowers: on the disc, between the petals, and between the sepals. The basic number of glands is 15, 5 in each cycle, although a variation of this number may be present. Observations indicate that there is as much if not more glandular surface in pistillate flowers as in staminate flowers. 4. Micro-sections show that the glands of both staminate and pistillate flowers have the same structure. The glands are devoid of conduction ele- ments, the secretion is exuded through the glandular mass and exposed upon the surface. 5. Tests indicate that the odor arising from pistillate flowers is as great or greater than that arising from staminate flowers. Literature Cited Bailey, L. H. 1935. The Standard Cyclopedia of Horticulture, Vol. I, fey Baril Britton, N. L., and Brown, A. 1913. An illustrated flora of the North- eastern United States, Canada, and the British Possessions, Vol. II, p. 446. Chamberlain, C. J. 1930. Methods in plant histology, 4th ed., p. 21. Green, C. H. 1939. Trees of the south, p. 324. Jepson, W. L. 1923. A manual of flowering plants of California, p. 606. Rehder, A. 1927. Manual of cultivated trees and shrubs, p. 527. Zirkle, C. 1930. The use of n-butyl alcohol in dehydrating woody tissue for paraffin embedding, Science, Vol. 71, pp. 103-104. RaeT as q TRANSACTIONS | ' OF THE ACADEMY OF SCIENCE (Quarterly Series) \ AFFILIATED WITH THE A.A.A:S. { Nn a VOLUME 11-NUMBER 2 . SEPTEMBER, 1943 THIRTIETH ANNUAL MEETING : 1943 CONTENTS 7 Page | Colonies of Bacterium Tabacum on Roots of Wheat and Several Grasses Stephen Diachun E. M. Johnson AYA 2g U9 ee Ne eee NM ah OUI LAS 17 ; Localization and Cleavage of Cytoplasm in Certain | Parasitic Protozoa ; } 1 igs 6s Oyo U el oT MM PN I ate frsceh cht catnanu ily So ae 20 Excretion of Manganese by the Rat J. T. Skinner a 4 aes ww VRGET aie Ue 5.) aS ascent EO NN IS ae : Bottom-Preferences of Fishes of Northern Kentucky Streams W. R. Allen Dvir seks Gear 24.056) i ALT Ng aa ep) Wale Wa 26 . PSTATACTS HME RENO E HE INAS 0 tl Ae aM MAO AA ICH Le Pn 31 ) ee es ee ey TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE . EDITORIAL STAFF Lawrence Baker...... Berea College...... Psychology and Philosophy Be Wi Coak ect cc aus, Centre Collesen aa scare Bacteriology Eb Bevel diee University of owmisvalle.\:\: 5. arene Biology ALG) McBarlanii. ©. University ofixentucky.)../ 0) aoe Geology Ward C. 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Remittances and correspondence con- cerning subscriptions, extra: costs, and other financial matters except reprints should be addressed to John Kuiper, University of Kentucky, Lexington, ae ar 709% Lt A ay aAc\\N\ WA eat Thirtieth Annual Meeting 17 COLONIES OF BACTERIUM TABACUM ON ROOTS OF WHEAT AND SEVERAL GRASSES*+ STEPHEN Diacuun, E. M. JoHnson, and W. D. VALLEAU Kentucky Agricultural Experiment Station Wildfire and angular leaf spot, common and often destructive bacterial diseases of tobacco in Kentucky, have been difficult to con- trol. Until recently the origin of the bacteria that initiate epidemics was unknown. But within the last two years, in the search for the source of inoculum, the bacteria have been recovered from overwin- tered field soils naturally contaminated from a previous infected crop (1), from the roots of wheat and other plants growing in natur- ally and artificially contaminated soils (2), and from the roots of inoculated plants of several species (3). The object of this report is to show that [1] Bactertum tabacum which causes the wildfire leaf spot of tobacco can grow and produce colonies on wheat roots inoculated under aseptic conditions; [2] the bacteria seem less able to grow on roots of some grasses, such as redtop, timothy, and Kentucky bluegrass. Method Wheat seedlings were used to study the growth of Bactertum taba- cum on roots because wheat roots are known to harbor the bacteria, and because wheat seedlings are easily and readily grown under aseptic conditions. Bluegrass, redtop, and timothy were also tested. The seeds were surface sterilized by soaking 2 hours in a 114 per cent solution of NaOCl; then germinated on sterile moist paper towel in sterile petri dishes. When roots were formed (2 to 7 days), they were dipped into an aqueous suspension of cells of Bacterium taba- cum, one to two million cells per cc. After inoculation the plants were so placed on sterile moist paper in sterile petri dishes that the roots were upright in the moist air, not in contact with paper or glass. From 2 to 5 days later the roots were examined microscopically on sterile slides. In some tests after the microscopic examination tobacco leaves were inoculated with the roots crushed in water. * The investigation reported in this paper is in connection with a project of the Kentucky Agricultural Experiment Station and is published by permission of the Director. 7 Presented at the 30th Annual Meeting, April 24, 1943, Louisville, Kentucky. 18 The Kentucky Academy of Science Results The bacteria consistently grew well on wheat roots. Usually colonies were present on the roots 2 days after inoculation. The colonies usually were most abundant on the central part of the root, i. e., the portion which was the root tip at the time of inoculation; the bacteria did not spread forward with root growth. Some of the colonies were strikingly tenacious. The one shown in figure 1 was kept mounted on a slide in water at 40°F and retained its shape for 5 days, with no visible diffusion of bacteria away from the colony. Whenever tobacco leaves were inoculated with crushed roots with such colonies on them, severe infection invariably resulted. (Table 1.) Roots of Kentucky bluegrass seemed less suitable for colony for- mation. With inoculum containing 2 million bacteria per cc. colonies developed on only an occasional root. When the bacterial count of the inoculum was increased to about 20 million cells per cc. colonies formed on 20 to 40 per cent of the roots. On roots of ryegrass, timothy, and redtop, more colonies were formed than on bluegrass, but fewer than on wheat. A summary is presented in ‘Table 1. Macroscopic and microscopic examination of the unstained living roots showed no evidence of injury, even near the bacterial colonies, up to 5 days after inoculation, when observations were discontinued. Discussion Since the discovery of the causal agent of wildfire of tobacco, more than 20 years ago, no satisfactory explanation of the sources of inoculum has been advanced. In our investigations we have found that the bacteria can be recovered from soil and from roots of several species of plants growing in fields in which infected tobacco grew the ‘TasLeE |. Formation of colonies by Bacterium tabacum on roots of seedlings. Wheat Ryegrass Timothy Redtop Bluegrass Number) (Of: (GESts siemens 6 2 4 3 3 Total number plants inoculated_......_== 96 40 80 50 40 Number plants on which colonies developed... 85 13 18 7 7 Number roots tested by inoculation onto tobacco: (leaves. sce a ee 40 40 60 15 40 Number roots that produced wildfire On tobacco, sae ee eet 40 il'7/ 14 5 7 Number non-inoculated control plants... 66 20 60 50 20 Number control plants on which colonies GLeviel ope hy ie 2c SI ree 1 ja 3a 7a 3a Number control roots tested by inoculation onto tobacco leaves — 20 20 20 10 10 Number control roots that produced wildfire on tobacco —_.-.-------------.- 0 0 0 0 0 a Obvious contamination; colonies not typical. Thirtieth Annual Meeting 19 Ficure 1. Colony of Bacterium tabacum on a root of wheat seedling. Root inoculated 2 days after germination. Photographed 2 days after inoculation. Root mounted in water. X 200. previous season. ‘The bacteria can grow and produce colonies on the surface of roots of wheat and several grasses, and thus can maintain themselves between crops of tobacco. It may be that this organism is not primarily a tobacco pathogen at all, but rather a common bac- terilum present on roots of many species of plants, which under special favorable circumstances can cause a leaf spot of tobacco and other hosts such as tomato, Physalis, and morning glory (Ipomoea). Perhaps some concepts of host range, and control of plant diseases by crop rotation should be reconsidered in view of the fact that some plant pathogenic bacteria can live and overwinter on roots of several unrelated species of plants. References 1. Diachun, Stephen, W. D. Valleau, and E. M. Johnson. Isolation of Bac- lerium angulatum from overwintered field soil. Phytopathology 32: 2-3, 1912. Abstract. 2. Valleau, W. D., E. M. Johnson, and S. Diachun, Association of tobacco leaf spot bacteria with roots of crop plants. Science 96: 164. 1942 3. Diachun, Stephen, W. D. Valleau, and E. M. Johnson. Multiplication of tobacco leaf spot bacteria on roots of other species. Phytopathology 33: 3. 1945. Abstract. Since this report was first written the bacteria have been recovered from roots of several crop plants in the absence of tobacco. Phytopathology 34: 163-174. 1944, 20 The Kentucky Academy of Science LOCALIZATION AND CLEAVAGE OF CYTOPLASM ; IN CERTAIN PARASITIC PROTOZOA* H. B. Croucu Kentucky State College Frankfort, Kentucky In 1915, Kofoid and Swezy! published a report and figures on the changes of body form in Tritrichomonas augusta of the frog. ‘They pointed out that the organism was able to localize its body areas by the constriction of the pellicle. Later, in 1920?, these authors described a similar condition in Chilomastix mesnili, the human intestinal fla- gellate. In addition to their earlier observations, they saw a living specimen of this species eliminate a large rod bacterium from the posterior end by the constriction of the body and ultimate cleavage of the cytoplasm surrounding the bacterium. More recently, several investigators have made notes on similar conditions in parasitic pro- tozoa; but the most of them gave hardly more than casual mention to their observations. Some suggested, however, like Kofoid and Swezy, that this auto-merotomic process represented a mode of egestion or defecation. This phenomenon has been observed many times by the present author in living and stained parasitic flagellates from a variety of sources. The process has been followed rather closely in several species of Trichomonas, particularly Trichomonas wenrichi, and a species of Chilomastix, all intestinal parasites of the common woodchuck (Mar- mota monax). At the present stage of the investigation, it is not known precisely how such a cytoplasmic pinching-off process is accomplished. But the frequency of its o¢currence suggests that it is a highly significant func- tion in the well-being of the organism. . In Trichomonas, it appears that the axostyle and the chromatic basal rod facilitate the process along with the progressive constriction of the body pellicle. Foreign solid matter within the flagellate is usually contained within a vacuole surrounded by a rather firm vac * Presented at the 30th Annual Meeting, April 24, 1943, Louisville, Kentucky. + Now associated with Tennessee State College. * Kofoid, C. A., and Swezy, O. 1915. Mitosis and multiple fission in trichomonad flagel'ates. Proc. Amer. Acad. Arts and Sci. 20: 145-168. ? Kofoid and Swezy. 1920. On the morphology and mitosis of Chilomastix mesnili (Wenyon), a common flagellate of the human intestine. Univ. Calif. Publ. - Zool. 20; 117-144. Thirtieth Annual Meeting 7A | uolar membrane. The vacuole together with its contents is forced toward the posterior end of the organism by the flexion of the chro- matic rod over the axostyle. Upon reaching the extreme posterior end of the body, the cytoplasmic mass surrounding the vacuole is isolated further by constrictions of the pellicle. Finally, the restricted area is pinched off from the body. The process in Chilomastix is similar to that of Trichomonas, except the twisting and constricting motions of the Chilomastix appear to be the only propelling forces. Some flagellates cast off rather large globules in the same manner, which appear to contain no solid particles whatever. The nucleus does not appear to take any part in the process at any time. The above described process may be termed ‘somatomy’, since nuclear reorganization is not involved. Following final cleavage of the cyto- plasm in this manner, the organism is obviously smaller, but appar- ently healthy otherwise. Unlike many free-living protozoa, parasitic flagellates have no well defined cytopyges. Neither do they eliminate solid matter by the rupture of the pellicle like many amoeboid protozoa. Many kinds of flagellates egest solid matter through the cytostome. However, the intake apparatus in some species is not well adapted to regurgitation. This is especially true of those having rather rigid cytostomes. In such forms, the rupture of the pellicle or cytoplasmic cleavage would afford the only alternatives for egestion. Somatomy may be regarded at least as an emergency function in the well-being of the organism, in the absence of other facilities for eliminating large quantities of foreign matter or metabolic wastes. Since the rather common hyper-parasite, Sphaerita, is often elim- inated in this manner, somatomy may by considered as a method of overcoming hyper-parasitism, along with its other egestive functions. The apparent homogeneous globules of cytoplasm are usually given off by the larger flagellates. This is highly suggestive that the metabolic load is materially reduced along with the reduction of the size of the organism. If this assumption is correct, somatomy accom- plishes cytoplasmic rejuvenescence, similar to the outcome of binary fission. From the above observations, it may be concluded that somatomy, or the casting off of cytoplasm, accomplishes the functions of the cytopyge in other protozoa, and cytoplasmic rejuvenescence in the absence of mitotic cell division. 22 The Kentucky Academy of Science EXCRETION OF MANGANESE BY THE RAT*+} J. IT. Skinner and J. S. MCHArGuE Contribution from the Department of Chemistry of the Kentucky Agricultural Experiment Station In connection with their studies of manganese metabolism in the rat Skinner, Peterson, and Steenbock (J) found that although the greater part of the element is eliminated in the feces, an appreciable amount is excreted in urine. For example, when rats were fed a ration low in the element, almost 20 per cent of the total manganese output was found in urine, but the proportion dropped to approxi- mately one per cent when the manganese content of the ration was increased forty-fold. Perla et al (2, 3) found that manganese retention was influenced by the diet employed and particularly by thiamin intake. That their second paper contains erroneous data on mangan- ese excretion is suggested by the tremendous retention, as calculated from intake and output, of 303 mg. of a total of 789 mg. of manganese ingested by each rat between April 12 and July 4. Kent and McCance (4) observed that in man only a very small part of food manganese was excreted in urine. Their conclusion “that Mn is not excreted freely, if at all, by the human bowel when it is injected in the amounts used for the experiment” is based upon balances admitted by the authors to be “somewhat irregular and unconvincing.” In view of these somewhat conflicting reports it was deemed desir- able to repeat the experiment on excretion of manganese at two dif- ferent levels of intake, and to include in the investigation a study of the effect of injected manganese upon elimination of this element by the rat. Procedure The adult male rats used in these studies were housed in indi- vidual screen wire (14 inch mesh) cages provided with false bottoms of the same material. Each cage rested in an enameled pan which was provided with filter paper for absorption of urine. A 14 inch mesh screen was placed above the filter paper to catch the feces and thereby prevent their continued contact with urine. * The investigation reported in this paper is in connection with a project of the Kentucky Agricultural Experiment Station and is published by permission of the Director. j Presented at the 30th Annual Meeting, April 24, 1943, Louisville, Kentucky. Thirtieth Annual Meeting 29 Feces were collected daily and urine papers were changed midway and at the close of each 6-day metabolism period. Following com- pletion of a period the false bottom of the cage, the feces-retaining screen, and the enameled pan were rinsed with distilled water and the washings were saved. The filter papers were first dried sufficiently to permit ready removal of bits of hair and any feed which was spilled during the period. All papers used with an animal during a given period were then placed in a beaker where they were extracted first with the wash water previously mentioned and then successively with four portions of boiling distilled water. The extracts were combined and after the resulting solution had cooled, it was diluted to a volume of 500 ml. for replicate sampling. Feces, spilled feed, and extracted filter papers were also analyzed for manganese. Most of the analyses were made according to the method of Skinner and Peterson (5), chlorides being removed by cautious heating after addition of phos- phoric acid. As a check on the efficacy of the phosphoric acid extrac- tion, a few samples were analyzed by the longer procedure as given in Methods of Analysis of the Association of Official Agricultural Chemmists (6). All color comparisons were made with a Klett-Sum- merson photoelectric colorimeter with the green filter, No. 54. The ration fed during Periods | and 2 was a commercial dog food! upon which the experimental animals were reared. Since a relatively high concentration, 75 mg. per kilo, of manganese in this ration made it unsuited to a study involving further additions of the element, a second ration was compounded which contained only 7.4 mg. of manganese per kilo. The latter was fed during the rest of the experiment and contained the following ingredients: per cent GOrnemeal screws eu) eran eee 78 GaSe Bes oe een AS Sirhan ae aaa nee L2 S Aili Smee tetas eis. Gcucdeatos Memento 4 BREWED SAV CASE tah a. nc! ean oe aaa 4. Codthivervoilk sy sss Soe ee 2 Each ration was fed ad libitum. A water solution of manganese lactate was injected subcutaneously on the second and fourth days of each of two experimental periods. Each injection was equivalent to 3.8 mg. of manganese. +Purina dog chow. 2A The Kentucky Academy of Science Results Urinary manganese as given in Table 1 is the algebraic sum of three values: [1] manganese transferred to aqueous solution by extrac- tion of urine from filter papers, [2] manganese remaining in papers in excess of that originally present, and [3] manganese absorbed by spilled feed. It will be observed in Table I that during Periods 1 and 2, when the manganese intake per period was approximately 8 mg. per animal, the urinary output of this element amounted to about 2 per cent of the total output. This small value was to be expected since the man- ganese content of the ration in comparison with that of most feeding stuffs was relatively high. When the high manganese ration was replaced by one furnishing but little more than one-tenth as much of the element, there was a marked drop in fecal manganese and a small but perceptible drop in urinary manganese. (See Periods 4, 5 and 6.) The net result was a great increase in the contribution of urinary manganese to the total output; the average for the three peri- ods being 13.4 per cent as compared with 2 per cent on the high man- ganese ration. This is somewhat lower than the value reported pre- viously (J), but it should be mentioned that.an intake of manganese averaging 12 per cent greater than in the former studies would tend to raise fecal output of manganese and hence minimize the contribu- tion of urinary manganese toward the total output. Positive balances during these periods doubtless were due to experimental errors and should not be construed as indicating storage since it has been ob- served (J) that adult rats contain less than 0.1 mg. of manganese when reared on a ration containing 10.47 mg. of the element per kilo. In Column 2 it will be noted that food manganese dropped appre- ciably during Periods 7 and 8. ‘This drop was due to a marked de- crease in food consumption which resulted when manganese injec- Taste I. Excretion of Manganese (Average of three rats) Period : Intake Output Part of (6 days) output Balance By mouth Injected Feces Urine in urine mg. mg. mg. meg. per cent mg. 1 8.13 8.24 0.16 1.9 —0.27 2 7.87 7.75 0.17 2.1 —0.05 3 0.82 1.47 0.13 8.1 —0.78 4 0.82 0.51 0.09 15.0 -+0.22 5 0.88 0.68 0.12 15.0 +0.08 6 - 0.89 0.61 0.07 10.3 +0.21 71* 0.53 7.6 4.65 0.09 1.9 +3.39 8 0.64 7.6 6.00 0.09 1.5 +2.15 g* 0.88 2.17 0.08 3.6 —1.37 * Average of only two rats. Thirtieth Annual Meeting 25 tions were initiated. Rapid loss of weight ensued with the result that during the twelve days constituting these periods the animals lost an average of 49 g. Total manganese intake as represented by that con- sumed in food and received by injection, however, approximated closely that of the first two periods when the high manganese ration was being consumed. Increased elimination of manganese in the feces lowered the percentage of urinary manganese approximately to 2 per cent of the total, a proportion obtaining in the first two periods. Failure to make a quantitative injection of manganese into one of the three experimental animals was responsible for including data on only two rats in Period 7. With regard to the apparent storage of more than 5 mg. of manganese per animal during Periods 7 and 8, it should be stated that each animal acquired one or more abscesses from the four injections of manganese lactate. It seems likely that the manganese in such areas would have been largely immobilized and hence its excretion would have been greatly delayed. Data for Period 9 substantiate this belief since a negative balance of 1.37 mg. occurred in spite of the fact that the final injection was made two days prior to the end of the preceding period. Assuming that injected manganese does not appear in urine, one would have expected the urinary output of manganese in Periods 7 and 8 to be somewhat less than in the three periods which preceded because, according to Column 2, food manganese was only 68 per cent of that consumed during the low-manganese periods. Since there was no decrease in urinary manganese, it appears quite likely that a part of the injected manganese appeared in the urine. Conclusions ]. On a ration containing 75 mg. of manganese per kilo, rats excreted 2 per cent of the element by way of urine. 2. Urinary manganese increased to 13.4 per cent of the total out- put on ration containing 7.4 mg. per kilo. 3. Manganese injected subcutaneously was eliminated readily by way of the bowel. Bibliography 1. Skinner, J. T., Peterson, W. H., and Steenbock, H., J. Biol. Chem., 90, 65 (1931). 2. Sandberg, M., Perla, D., and Holly, O. M., Proc. Soc. Exp. Biol. and Med., 42, 368 (1939). 3. Perla, D., Sandberg, M., and Holly, O. M., Proc. Soc. Exp. Biol. and Med., 42, 371 (1939). . Kent, N. L. and McCance, R. A., Biochem. J., 35, 877 (1941). . Skinner, J. T., and Peterson, W. H., J. Biol. Chem., 88, 347 (1930). . Official and ‘Tentative Methods of Analysis of the Association of Official Agri- cultural Chemists, Washington, 5th Edition, 1940, 128. Dore 200Gae The Kentucky Academy of Science BOTTOM-PREFERENCES OF FISHES OF NORTHEASTERN KENTUCKY STREAMS* W. R. ALLEN and Mrtnor E. CLark Department of Zoology, University of Kentucky, and Kentucky Division of Game and Fish, Frankfort During some three summers the junior author was engaged in stream-survey work for the Kentucky Division of Game and Fish. The principal areas covered were the basins of the Big Sandy and Little Sandy Rivers, Tygart’s Creek, Kinniconnick Creek, and the Licking River, all in the northeastern section of the state. In con- nection with other features of the project, he, together with the senior author, gave considerable attention to the matter of local distribu- tion of fishes. This emphasis was fruitful of a rather voluminous accumulation of data with respect to the types of stream-bottom upon which fishes were collected. We have 945 records of the occurrence of particular species upon specific types of bottom in given localities. From these it seems reason- able that some conclusions may be drawn as to whether the fishes in question do exercise choice of bottom, and if so, where their pret- erences lie. . We recognize seven distinct types of stream-bed, which, with their respective intergradations and combinations, give us a total of twenty-five discernible varieties of bottom. Each of our records means the occurrence of one species on a certain type of sub-stratum in one locality. ‘That is our unit for calculation. If five species or ten species were taken together, they are entered five or ten times correspondingly in the record. Each species is entered into the record once for each occurrence in a different type of bottom or different locality, in the same stream or different streams. No account is taken of the numbers of individuals collected. The seven principal bottom-types were: (1) Bedrock, (2) Boulder, (3) Rubble, (4) Gravel, (5) Sand, (6) Mud, and (7) Detritus. It will readily be seen that this series is arranged in accordance with the coarseness or fineness of the material; also that they represent a grada- tion of materials correlated with the velocity of the current, the bed- rock having been transported not at all, the boulders but little with the assistance of steep gradients, the rest grading out to a point at which detritus occurs in the sluggish pools and backwaters, far downstream. Due to considerable declination of the terrain within the east- * Read at the Thirtieth Annual Meeting, April 24, 1943, Louisville, Ky. Thirtieth Annual Meeting 7a west axis of the state, something on the order of 4000 to 400 feet in 400 miles, our streams possess considerable gradient, much moderated, however, by the fact that they are deeply entrenched toward their headwaters, and much of the fall assumed by their numerous and declivitous tributaries. Thus they present problems in the dis- tribution of aquatic life which would seem to set off the region from the Gulf and Mississippi lowlands on the one hand, and from an aggraded glacial terrain on the other. The present paper looks toward a more complete future appraisal of the problems of dis- tribution. A chart, Table I, shows the types of correlation existing between current velocity and bottom structure, measured in terms of the num- bers of species collected from each. From this you will note that at “one extreme we have eight records from bedrock bottom, all showing swift water. At the other extreme no fishes were recorded upon a bottom of pure detritus, and only five records where detritus even existed in combination with sand and gravel. Only 18 occurrences are attributed to mud bottom, all in sluggish water, and eight in situa- tions even partly muddy, in water mostly sluggish. Boulder-bottoms Tas_e I. Correlation of Bottom with Velocity of Current : Current Relative ; fineness : Type of bottom Moderate Sluggish Swift of bottom Rec- Per Rec- Per Rec- Per Total materials ords cent ords cent ords_ cent 1 DE YS (6 FXG) A eae Se ul Siete PV poke aE Ee oA capa 8 100.0 8 IEE Pee Bedrock, Rubble, Gravel _... 1 16.6 3 50.0 2 33.3 6 ss 3 Bedrock, Rubble, Sand _._ 1 33.3 2 GES Glee A ee 3 1, 3, 6 Bedrock, Rubble, Mud ____... 1 TOOL OS oho eae yay eee ae ee aa eek 1 1, 4 Bedrock, Gravel elise 3 21.4 11 718.6 14 15.455 Bedrock aGravel) cand) 2222 4 80.0 1 20.0 b) ihe 5} Bedrock, Sand _._..__ 1 20.0 2 40.0 40.0 5 1, 6 Bedrock, Mud _...___ ee 1 LL OOE Oy ere et ig Spee ia heey mnt PUT 1 2 Boulders = eee EEN a age eee Re a Exes 1 100.0 1 2, 3, 4 Boulder;> Rubble, ‘Gravel. —— =~) =e ee 1 100.0 1 3 Rub ble eae Fa Nos ee 1 TA pe epee 6 85.8 7 ane. Rubble, Gravel __...__ 2 2.8 9 12.7 60 84.5 71 ay ee ts) Rubble, Gravel, Sand _ a 2 P78} 43 48.3 44 49.4 89 By) Rubble, Sand —.-. SO pk iy Se pee 5 71.4 2 28.6 th ae BL G Rubble, Sand, Mud py eee Vp ee 1 NOOO ato) eee 1 4 Go AVG ls) ste ate 39 32.8 44 36.9 36 30.3 11S 4,5 Gravel, Sand 27 7.5 249 69.2 84 23.3 360 4, 5, 6 Gravel, Sand, Mud __.. 3 LOO IO ya ocean a ae ne ene Pay = ek Ay ate 8 3 4,5, 7 Gravel, Sand, Detritus 2 66.6 1 SO vOlen ye) UE eh Gish 3 4,6 Gravel, Mud _.....__ = 2 TOO: Oe Ree Wt ates, tae ST yh 2 5 S211 Clee ee ava é 42 24.7 124 73.8 3 1.5 169 5, 6 Sand, Mud __ 45 91.8 4 SiO erie LU Rees 49 ay Ul Sand, Detritus & Deh ghee ESS Ree oe DA ee ee ea 2 6 IMA iy ee as 18 ‘TOO One ee pee ere hn Sel ge ee 2 18 7 GLEE UG pee a is eee ee Ieee aed oe ah pee oe ee aie ea et, PR OGA Siege te 190 20.1 494 52.3 261 27.6 945 The degrees of fineness or coarseness of the bottom materials are expressed as numerals from one to seven, ranging through bedrock, boulder, rubble, gravel, sand, and mud to detritus. (First column.) The numbers in the columns headed ‘‘Records’’ indicate the frequency with which distinct species were taken on each type of bottom, in as many distinct localities. 28 The Kentucky Academy of Science are exceedingly uncommon, and have yielded only one record, with one record also in combination; both were in swift water. The rubble bottoms, nearly all in swift water, produced seven species-records; when in combination with other materials, we have 179 occurrences of fishes on mixed-rubble bottoms, of which 109 were from swift water, 63 in moderate, only 7 in sluggish. Upon gravel, we have 119 records, almost equally divided among sluggish, moderate, and swift water. Upon sand bottom we have 42 records in sluggish water (fewer than in gravel), 124 in moderate current (three times the record on gravel); only three in swift current on sand. The total for sand, 169, as against 119 for gravel, may be accounted for in part by the greater predominance of sand from the weathering sandstone of the region. The chart indicates clearly that a much higher frequency was ob- tained upon mixed gravel-and-sand bottom than any other, the total of 360 records being 38 per cent of the grand total of all records. This may be compared with the occurrence of 52 per cent (494) in mod- erate current, as against 20 per cent (190) in sluggish water, and 27 per cent (261) in swift current. In the region in question we have some 98 species of record. Welter has reported 70 from the Licking River drainage, Clark 52 from the Levisa Fork of the Big Sandy River. Of the 98 known species, our record applies to 74. With some allowance for taxonomic revision, we find 21 species and subspecies taken on bedrock bottom, 39 on rubble, 66 on gravel, 67 on sand, 36 on mud, and 5 on detritus. Thus not only are the records for sand and gravel much greater in number than for other substrata, but a much larger number of species are found frequenting those types of bottom. Of the total number of combinations of different species with each of the types of bottom on which it had sometimes been found, and casting out duplicates, we have 236 distinct from one another, and 133 of this total, or 56 per cent, occur where sand or gravel is present. In ‘Table II we have an analysis by species of certain representa- tive distribution records, about one-third of the total, and enough to explain how the totals were arrived at. This table also shows, of course, the great predominance in the columns headed sand and gravel. However, Table III, a summary by families, may Puseae the total picture more graphically: With no significant exceptions, the preferred habitat of this fish population is upon bottoms in which sand and gravel predominate. This may be due either to the actual choice of a preferred substratum, or to the selection of waters having a preferred velocity, which hap- . pens to correspond with the resting-state of sand and gravel. The Thirtieth Annual Meeting 29 Tas_e II. Distribution of a Selected List of Species Types of bottom Check Ole uray 4 Our list Species gf s 3 > cs} Gs We SG Serie) oa ish aa eae is oe & O wo 2) fa) 10 7136 Catostomus commersonii 2 3 11 11 2 re 11 7156 Hypentelium nigricans E 6 25 38 1 1 15 781 Moxostoma erythrurum __ sie eas 7 10 2 as 18 800 Chrosomus erythrogaster Cee Crneaeea Rees 1 4 6 2 1 19 833 Semotilus a. atromaculatus 3 1 19 49 93 3 is 23 880 Notropis photogenis ae e 1 11 8 a2 pune 24 895 Notropis rubellus Eee tig) B 6 15 19 bak Fe 26 904 Notropis u. umbratilis Pee Meh a ES 8 5 2 27 905 Notropis ardens lythrurus 1 ae, 1 3 3 sais 29 934 Notropis cornutus chrysocephalus _._ 3 8 36 40 2 30 940 Notropis spilopterus 2 3 10 9 1 31 (940) Notropis whipplii pad 0 6 18 18 aes ! 32 976 Hybopsis amblops amblops ____ 1 coat 12 14 1 35 993 Notropis deliciosus stramineus eeu rao) 3 20 23 1 37 1022 Nocomis micropogon _—_._.._.-. 1 = 3 11 10 a) 42. 1063 Ericymba buccata Po oN ate 2 1 12 22 4 45 1091 Hyborhynchus notatus —__. vesanssal baie 23 46 43 1 47 1098 Campostoma a. anomalum __._..____ 4 1 19 41 49 2 52 1164 Ameiurus nebulosus nebulosus _.___ 1 3 2 4 58 1182 Schilbeodes miurus —_._.._- Wi oe a he 10 11 2 s 62 1974 Labidesthes sicculus — Sel: a 4 8 " 3 64 2185 Percina caprodes caprodes Eee ee a pe 13 9 il 65 2190 Hadropterus maculatus __.. WAST} 11 38 43 4 71 2217 Etheostoma b. blennioides - aaa 8 16 10 pee Z 713 2225 Boleosoma nigrum nigrum - 1 3 15 27 3 2 80 2276 Catonotus f. flabellaris ___ 3 = 1 17 16 1 e 26 2324 Lepomis auritus " s 1 8 9 2 S 89 2329 - Lepomis megalotis megalotis SEU ees 3 6 24 29 3 ae 93 2351 Pomoxis annularis —__---.------- 1 2 4 2 2 be Total individual species (duplicates not included) —.--.--W2.- 21 2 39 66 67 36 5 Total records (including duplicates) —_._..... 40 2 178 667 #&673 714 5 For purposes of analysis bottoms of mixed character are counted under all the heads, as, for example, a sand-gravel area would be included under both sand and gravel. the totals in this table do not tally with those in Table I, and we have a grand total of 1639 as against 945, Table I. Check-list numbers based on Jordan, Evermann and Clark, 1930. TaseE III. Distribution by Families Hence Type of bottom Families f Coarser Gravel Sand Finer Catostomidacy == 9 49 63 7 Cyprinidae __ _. 126 344 367 36 Ameiuridae La 7 26 22 10 Esocidae __... a eee 3 3 eu Cyprinodontidae . ib deme neees aM Yin ets WNC o es 3 3 Percopsidae) 222 ee si 2 4 3 He Atherinidde ee ee a 6 8 7 3 Etheostomidae _. ft 59 182 173 10 Gentrarchidae 2225 10 45 49 10 30 The Kentucky Academy of Science latter choice would be in the middle range of swiftness. No matter which motive actuates the fishes, we do find a definite correlation at any rate. Our present analysis is out of balance from the fact that few of the data were obtained from deeper water, such as the Ohio River and lower courses of its tributaries. Depth may be a factor which would largely cancel out any bottom selection, except for those species frequenting the bottom. This unbalance of our record is reflected in the small numbers of certain families such as catfishes or the Esocidae in the record. The record is, then, chiefly that of the more settled time of year; we offer no data as to what happens during freshets or the occasional excessively dry season. In further interpretation of the data, we can well understand cer- tain reasons for the paucity of records in the extreme situations. Our collections came mostly from shallow streams, and from the more shallow portions of those streams. ‘Therefore the bed of the streams would have the maximum effect upon the life of the fishes. The effect of detritus and mud upon the gills should be found at their greatest, and that type of bottom should be avoided to a greater extent than in deep water. Our lack of specimens from boulder- strewn waters is the common experience of those who do collecting in all kinds of streams. Furthermore, we should expect in sandy or gravelly situations the greatest amount of plant growth, such as Pota- mogeton, and in rocky or boulder-riffes only the minimum. ‘This should affect the amount of food and shelter in favor of the sand- gravel complex. Bedrock is found more often well upstream, in sections most exposed to the flash-freshets of our climate. ‘This fact, together with a probable sweeping out of food-supplies, renders the well-swept floors less acceptable. Rock-bottom pools further down- stream have a decided attraction for some species. ‘The question might be raised whether the use of a current-meter to determine velocities and screens for bottom-materials would elim- inate the personal factor. However, for purely qualitative purposes, we believe we have been more than reasonably consistent. Our total records of 667 for gravelly situations, 673 for sand, added together give 1340, or 82 per cent of all records. ‘This should be significant, with ample allowance for error. Thirtieth Annual Meeting 31 ABSTRACTS Field strains of tobacco mosaic.*+ E. M. JoHNson and W. D. VAtL- LEAU, Kentucky Agricultural Experiment Station.—Field and green- house inoculation studies with 54 tobacco mosaic strains, collected during 15 years in Kentucky tobacco fields, resulted in a wide range of symptoms, distinct for each strain, on necrotic spotting varieties (N’) of Burley and dark tobaccos. Inoculated plants developed various shades of mottling, from dark green through yellow to pure white, accompanied by varying degrees of dwarfing, necrosis and distortion. No two strains caused identical symptoms, either in the greenhouse or field. ‘This study indicates that many strains of the tobacco mosaic virus exist in nature. The suggestion is made that the terms “Tobacco virus 1,” “Common field tobacco mosaic” or “Wild-type tobacco mo- saic’ should be avoided in scientific papers when used in the sense that this is sufficient description for identification, because any of the 54 strains used in this study is a common field or wild-type strain, yet no two strains are identical. ~ The influence of nutrition on sporangial formation in Araiospora streptandra.? RicHARD Curtis WessTER, School of Medicine, Univer- sity of Louisville—The discovery of Araiospora streptandra in Ken- tucky is a new record for the genus. Hitherto it has been found only in Massachusetts, Rhode Island and North Carolina. A pure culture was obtained, facilitating the study of the effect of nutrition on sporangial formation. It was found that spiny spor- angia were formed under favorable nutritive conditions, while smooth sporangia were formed as a result of drastic reduction of nutrition. Smooth sporangia discharge their zoospores in a shorter time than do the spiny sporangia, the former being the chief means of dispersal to a more favorable substratum. Cytogenesis within the suwpra-optic and paraventricular nuclei of the pig.t Paut Roore, School of Medicine, University of Louisville.— In a series of 32 pig embryos and fetuses ranging from 15 mm. to term the following cellular changes occurred within the supra-optic and paraventricular nuclei: there are two stages in the development of the cells, the first stage somewhere around mid gestation and the sec- ond a few days previous to parturition. These two stages may be called growth periods. They are so designated because of an apparent * The investigation reported in this abstract is in connection with a project of the Kentucky Agricultural Experiment Station, and is published by permission of the Director. + Read at the 29th Annual Meeting, April 11, 1942, Lexington, Kentucky. + Read at the 30th Annual Meeting, April 24, 1943, Louisville, Kentucky. 32 The Kentucky Academy of Science sudden increase in size of the cell bodies of the neurons. The density of the vascular bed around the nuclei is correlated with the increase in size of the neurons. There is no evidence of endocellular capillaries “nor the presence of multinucleated cells as described by various inves- tigators for the adult forms of other vertebrates. ‘These same investi- gators have also reported peculiar secretory activity within these hypothalamic nuclei but cytogenesis of the cells within the pig gives no evidence of such phenomenon. All other nuclear and cytoplasmic changes are no different from the cells found in other diencephalic nuclei. Spinal ganglion hypoplasia after limb amputation in the fetal rat. E. K. Hatt and M. A. SCHNEIDERHAN, Departments of Anatomy and Chemistry, School of Medicine, University of Louisville.*—In a series of 45 cases, pregnant rats of the Nutrition Colony of Professor A. W. Homberger were operated on according to the technique of J. S Nicholas for amputation of the fore limb of the fetus. Such fetuses, fixed at term and sectioned serially, reveal a reduction on the ope- rated side in the size of the spinal ganglia of the brachial region and in the number of ganglion cells. One specimen, operated upon slightly less than 5 days before birth and fixation, may be described in detail; 11,910 cells were counted. The 8th cervical ganglion of the operated side shows the greatest reduction in cell number (79 per cent) as compared with the contralateral ganglion. ‘The reduction in the 7th cervical ganglion is less marked (34 per cent), and it is still less in the Ist thoracic (20 per cent). In the 6th and 5th cervical ganglia, the reduction is slight (8 and 15 per cent). No indications of injury or degeneration were found in the ganglia. ‘These results ex- tend to the mammal similar observations of Shorey and of Hamburger on the chick, and of Detwiler and others on the amphibia. The development of the uropygial gland in pigeons.* G. B. PENNE- BAKER, Morehead State Teachers College.—The uropygial gland in pigeons arises as two invaginations of the epidermis on the dorsal side of the uropygium, making its first appearance during the eighth _ day of incubation. The reservoirs form first, the secreting tissue orig- inating later as buds from the reservoirs. The lumina of the buds of glandular tissue do not appear until the gland begins secreting between the ninth and fourteenth days after hatching. Adult size of the gland is reached when the squab is about thirty days of age. In birds that do not possess the gland as adults, it does not begin development and later degenerate as has been suspected by some. * Read at the 30th Annual Meeting, April 24, 1943, Louisville, Kentucky. Pee ‘ 7 TRANSACTIONS OF THE . KENTUCKY - ACADEMY OF SCIENCE (Quarterly Series) AFFILIATED WITH THE A.A.AS. VOLUME 11 (Double Issue) No. 3—December, 1943 No. 4—March, 1944 THIRTIETH AND THIRTY-FIRST ANNUAL MEETINGS 1943 and 1944 CONTENTS The Root System of Azlanthus Altissima Weis SAVES UN Fe a facade ach ulate eS Ah) cage eas 33 Preliminary Studies on the Separation of Ether- Insoluble Pigments from ‘Tobacco Dirk Verhagen SHOT WM GEN! ie e282 Sih os wets pmo tee ane 36 Ontology of Consciousness KeSteno.O Bannon sj. 01) Bec. eee Seo NR aha Oe 41 Influence of Manganese Intake upon Urea Excretion J. T. Skinner Aeros EC Rar oUle Na ie Uo eae 47 Derivatives of 1-dimethylaminomethyl-2-naphthol Gerald dy "Grillo sats oe enn tegens eek Ren ir 51 Influence of Population Number on Egg Production in the Four-spotted Pea Beetle, Bruchus Quadrimaculatus Fabr. JAIUNWECOL AI BTEZRETS Channi PrN Enon hella Nes a ClM ten Mahe tea 56 FARMS GAG LSU RMI Pc (iota cg coke SIME e age Lica aIOt Se. 2 hens 63 ee ay A TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE EDITORIAL STAFF Bai Lawrence Baker...... Berea’'College> 74x Psychology and Philosophy — HEAVY, VOOR Lah i er Gentre Golleser ac as. ee eee Bacteriology PLB: ovelbai iah. « University of ‘Louisville. 2): ae. beeen Biology A. C. McFarlan...... University of Kentucky.............. Geology Ward C. Sumpter.... Western State Teachers College... .Chemistry Jamvass~ Wodd oor. rte 2). University of Kentucky .3: )o 34. Physics | John Kuiper..... University of Kentucky, Managing Editor Manuscripts. The Transactions must be limited to the proceed- ings of the annual meetings of the Kentucky Academy of Science and to original manuscripts pertaining to science. Manuscripts are sub- ject to the approval of the Editorial Staff and may be submitted to the Editor of the subject covered or to the Managing Editor. _ Extra-Cosr Features. The extra cost of special features such as cuts, graphs, tables, etc., above the text-run price per page must be borne by the contributor. The Editorial Staff will advise contributors concerning the extra cost of features upon receipt of manuscript. Illustrations to be included in an article should accompany the manu- script if possible, or, if sent in separate package should be properly: labeled as to the article in which they are to occur. . Proor. Galley proof will be sent for approval of contributors. The proof should be returned promptly to managing editor. Reprints. Reprints are furnished at publisher’s prices by nego- tiating directly with the printer. Price quotations on reprints are sub-— mitted with the proof, and orders for reprints should accompany the proof when returned to the printer. SUBSCRIPTION Rates. The Transactions is sent without additional expense to all members of the Kentucky Academy of Science who are not in arrears for annual dues. The annual subscription rate for non- members is $2.00 in the United States and Canada, $2.50 in foreign countries; single numbers 75 cents. One volume of four numbers appears each Academy fiscal year. : BusINEss CORRESPONDENCE. Remittances and correspondence con- cerning subscriptions, extra costs, and other financial matters except — reprints should be addressed to ene Lobates aera of Kentucky, — Lexington, Kentucky. SONIAN INS ae Ae LINN [s 329998 | iy 2 = O45 oe Uarong. yyseo— $] ee, AP"h y — i eal Z Thirtieth and Thirty-first Annual Meetings 33 THE ROOT SYSTEM OF AILANTAUS ALTISSIMA* P. A. Davies Department of Biology University of Louisville Although the extended distribution of Ailanthus altissima in North America indicates its tolerance to a wide range of habitats, certain growth conditions are more favorable than others for its maximum development. (/) Observations indicate that the most fav- orable growth conditions are: (a) open areas where there is little crowding by other plants, (b) deep, light soils containing an abun- dance of organic material, and (c) good drainage. Young plants are very sensitive to crowding and continuous shade. These conditions result in numerous spindly shoots which rarely live more than one year. Older trees which have their foliage above that of the surround- ing plants will tolerate crowding without apparent injury. Although Ailanthus will thrive on heavily packed clay soil of city streets to sandy beaches of almost pure sand, it develops more rapidly and to a greater size with a more luxuriant foliage on a deep loam soil with an abundance of organic materials. Good drainage is of paramount im- portance for its survival. The plant does not exist in swamps or live long on areas of poor drainage where water accumulates even for short periods. Native stands are found on high ground or where there is a lower drainage area. ‘The requirement for good drainage may be associated with a high oxygen requirement. Mangin made some studies on the relation of oxygen and carbon dioxide content in the soil to the opening of the buds in this plant. In buds which were open he found that the oxygen content of the soil varied from 17.86 to 20.30, with an average of 19.17 per cent, and the carbon diox- ide content varied from 0.35 to 3.43, with an average of 1.51 per cent, while in buds which were closed the oxygen content of the soil varied from 3.16 to 15.92, with an average of 10.96 per cent, and the carbon dioxide varied from 3.89 to 24.84, with an average of 9.5 per cent. (2) The drought resistance of Ailanthus is high. Stiles and Melchers ob- served that during 1934, which was the hottest summer recorded in Kansas, that 20 per cent of the trees examined were killed and another 30 per cent injured, and that young Ailanthus altissima trees in the nursery as well as old established trees were one of the trecs least injured by the drought. (3) The rapid rate of growth with a heavy sub-tropical foliage, its tol- erance to a wide range of soil types, and the ability to withstand severe droughts must make a considerable demand on the root system. Such * Read at the Twenty-ninth Annual Meeting, April 11, 1942, Lexington, Ken- tucky. 34 The Kentucky Academy of Science demands would indicate both a shallow spreading system for anchor- age and absorption and a large tap-root for water storage. As it 1s impossible to formulate rules in regard to the depth to which roots will penetrate, only digging will reveal what they actually do. To gain information on the root habits of this plant excavations were made on plants ranging from seedlings approximately two inches in height to ‘mature trees, the largest of which showed twenty growth-rings. Seedling studies were made in the laboratory. Seeds were planted in a good grade of greenhouse soil. When the plants were approxi- mately two inches in height the soil was washed from the roots by a gently flowing stream of water. Records were made immediately of the roois of these plants. Data from these studies show three general characteristics of the root system: [1] it is shallow and spreading, [2] the roots near the stem thicken into a horizontally enlarged storage structure, and [3] ad- ventitious shoots arise from the smaller roots of mature plants near the surface of the soil. The shallowness of the distribution of the root system is evident in all stages of growth. Even in mature trees the roots are almost entirely confined to the upper layers of the soil. Fig. 1 shows the horizontal distribution of the root system in a mature tree exhibiting twenty growth-rings. he root spread of this particular tree was uniform and extended approximately fifteen to twenty-five feet from the main stem. Fig. 2 shows that the root system is shallow and is chiefly confined to the upper eighteen inches of the soil. The deeper lying surface roots send numerous small roots to the surface. Adventitious shoots which arise from small surface roots of mature trees exhibit the shallow spreading root habits of the mother tree (Fig. 3). Seedling root systems show the same characteristics as those observed for mature trees and adventitious shoots. The primary and secondary roots spread out and grow horizontally in the upper inch of the soil (Fig. 4). ‘To compensate for the shallow distribution of the root system the larger horizontally distributed roots near the stem thicken into a large compact storage body (Fig. 2). The adventitious shoot which arises from the smaller surface roots of mature trees stim- ulates the mother root beyond the point of attachment to enlarge into a storage structure (Fig. 3). ‘The presence of the storage body may account for the drought resistance of Ailanthus observed by Stiles and Melchers. (4) As long as the mother plant remains active few adventitious shoots arise from its smaller roots near the surface. How- ever, when the dominance exerted by the mother stem is destroyed either by death or destruction many shoots arise from the extended reot system. ‘This situation results in a thicket of many, rapidly grow- ing shoots. (3) 2 6 8 {0 Distance In Feet ~ ai AS ON w : e Fig. 2 == a Distance In Feet Distance In Inches Fic. 1. The horizontal distribution of the larger roots in a mature tree with twenty growth-rings. Fic. 2. A cross-section of the vertical distribution of the larger roots in the same tree shown in Fig. 1. Fic. 3. Size and vertical distribution of the root system of an adventitious shoot which arose from one of the smaller roots of a mature tree. Fic. 4. Distribution of the primary and secondary roots of seedlings. Bibliography I. Davies, P. A., 1941. The history, distribution, and value of Ailanthus in North America. Trans. Kentucky Acad. Sci. 9: 12-14. 2. Mangin, L., 1895. Sur l’aeration du sol dans les promenades et plantations de Paris. Comp. Rend. 120: 1065-1068. 3. Stiles, E. H., and Melchers, L. E., 1935. The drought of 1934 and its effect on trees in Kansas. Trans. Kansas Acad. Sci. 38: 107-127. . Ibid. . Davies, P. A., 1935. Ecology of Ailanthus altissima thickets. Trans. Kentucky Acad. Sci. 7: 24. Or 36 The Kentucky Academy of Science PRELIMINARY STUDIES ON THE SEPARATION OF ETHER-INSOLUBLE PIGMENTS FROM TOBACCO* Dirk VERHAGEN} and Simon H. WENDER Contribution from the Chemistry Department, University of Kentucky For this study of the color-producing chemicals of tobacco, the pig- ments have been arbitrarily divided into two groups, based on their relative solubilities: the ether-soluble and the ether-insoluble pig- ments. The ether-soluble fraction, containing the chlorophylls and carotenes, has been studied for some time by other workers, such as R. N. Jeffrey and R. B. Griffith (/), at the Kentucky Agricultural Ex- periment Station. This present preliminary report deals with the separation and study of some of the ether-insoluble pigments. Experimental The method of separation involves extraction with proper solvents, partition between immiscible solvents, and use of chromatographic adsorption analysis. Using these methods, at least five of the ether- insoluble pigments of tobacco have been separated. Extraction A sample of lug Burley tobacco (Nicotiana tabacum) was obtained from the Kentucky Agricultural Experiment Station in Lexington. It was grown and cured on the Experiment Station Farm during the season of 1942. ‘This tobacco was stemmed and then powdered in a Wiley mill. Ten grams of the powdered tobacco were treated with 85 ml. of acetone and 15 ml. of water, and this mixture was thoroughly shaken and allowed to stand for at least 30 minutes. Fifty ml. more of water were added, and the mixture allowed to stand for about 5 minutes. The extract was then filtered by suction, and the residue washed with several small portions of a water-acetone mixture con- taining 55%, acetone. The filtrate was combined with an equal vol- ume of ether in a separatory funnel and mixed very carefully. The acetone-water layer was removed, and the ether layer was washed with 50 ml. of water, and this water was then added to the acetone-water extract. The aqueous layers were washed with five 50 ml. portions of ether. All ether portions were now discarded. The acetone-water extract should then be free of the ether-soluble pigments, and con- tain the portions of the ether-insoluble pigments studied in this inves- * Read at the Thirtieth Annual Meeting, April 24, 1943, Louisville, Kentucky. + Present address: Lyle Branchflower Co., Seattle, Washington. Thirtieth and Thirty-first Annual Meetings on tigation. ‘This extract was then made up to a volume of 250 ml. in a graduated flask with a water-acetone mixture containing 90% acetone. Chromatogram Chromatographic adsorption methods (2) were employed in the further separation of the extract. For this separation it was found that when 2 grams of Hyflo Super Cel were mixed with 10 grams of Alorco activated Alumina, which had previously been powdered so that all of it would pass through a 100-mesh screen and half through a 200- mesh screen, good resulting adsorption columns could be obtained. The adsorption column was formed by pouring an acetone suspension of this mixture of Hyflo Super Cel and Alumina through a glass col- umn about 1.9 cm. in diameter and 37 cm. in length and constricted at one end. A cotton plug had been placed in the constricted end of the column. The column was then washed thoroughly with acetone, afterwards with a few ml. of water. Twenty-five ml. of the ether-insoluble extract were diluted with 50 ml. of acetone and passed through the column. As the liquid passed through the adsorbant, most of the pigment material remained at the top as a uniform band of color, about | cm. in depth. The adsorption column was washed with acetone until the percolate no longer showed any color. ‘The adsorption column was then washed with large quan- tities of 80% acetone. As the 80% acetone passed through the adsorbant, three bands of color formed and moved down the column. Each was collected sep- arately. ‘The first pigment solution to be eluted was called pigment number one; the second, pigment number two; and the third, pigment number three. Pigments 1, 2, and 3 Each of the solutions was in turn purified and concentrated by re- adsorption on a different, smaller column; then eluted again. The re-adsorption was accomplished by adding a large volume of acetone to the pigment solution (2:1), and passing the diluted solution through the new column. With this dilution, the pigment was re-adsorbed at the top of the column. It could be then eluted with 80% acetone. Thus, a purified solution of pigment one; another of pigment two; and a third, of pigment three, were obtained. Pigment 4 When no further color could be eluted from the original adsorp- tion column after the first removal of pigments one, two and three, the adsorbant was washed with distilled water. A band of color moved down the column rapidly, and was collected and called pigment num- 38 The Kentucky Academy of Science ber four. It was purified by dilution with three volumes of acetone; re-adsorption on a new column; washing with acetone; then elution again with water. Pigment 5 After the removal of pigment number four from the original ad- sorption column and thorough washing with water, a band of color still remained adsorbed at the top of the column. This ‘color was eluted with 0.25% hydrochloric acid solution. At least two bands moved down the column. Each was collected separately. hese were called pigment number five and pigment number six. Further study of this separation is now in progress. Absorption Spectra To study the first five of the pigments obtained in the above pro- cedure, the absorption spectra curves were studied by means of a Cenco-Sheard spectrophotelometer (3). The curves were determined for wave lengths between 336 mz and 650 mv. A blank was prepared by using the same procedure as the method used in purifying each of the pigment solutions. However, instead of adding the pigment solu- tion to the column, the solvents of approximately the same concentra- tion were used alone. Pigment number one had one maximum, at 350 mv (4). This max- imum was quite sharp and no further maxima or minima were ob- tained for this pigment in the range examined (Fig. 1). Pigment one is yellow in color with a slightly golden fluorescence. Pigment number two had a sharp maximum at 34] me and a second slight maximum at 525 mz, (Fig. 2). This pigment was yellow with a pink fluorescence. Pigment number three exhibited a sharp rise towards a maximum just below 330 mse, a minimum at 375 me; and a maximum at 420 mez, (Fig. 3). This third pigment was yellow, with a greenish fluor- escence. Pigment number four showed a sharp maximum at 345 me (Fig. 4). Pigment number five had a sharp maximum at 352 me. Neither had other maxima or minima in the range examined. The curves for solutions of pigments number one; four, and five have only one sharp maximum point each. The values for these max- imum points, 350 mz, 345 me, and 352 mz, would indicate that the pigments, though different, are similar in type. Pigment number three is of a different type. It would indicate a maximum below 335 mz, and a second maximum at 420 mz, Further studies on these pigments are in progress. Vv aaunosis ; 2 aunois Unoda LNAWIId WOd AAUWND VULOAdS Nolldwosav SOMATA, i TELLONGACaNUA OML LNAWSId YOs SAUND VULoaAdS NOlLdwdosaVv er) 009 00S oor sve o2zc SNOUSINITTIW NI HLON|T aAWM oF = Koll 009 00s oov Ive O7e 21 oz “a vi oc Le 91 ov roa t aunoi4 SNO LNAWSId dOs SAUND VdLOAdS NOILdYosav SNOXOIIWITTIW Nt HLON3AT SAVM € saydNnol4 009 oos oor oss o2e SaYHL LNAWSId YOs SAWN VaLDAdS NOILdyossv SNOXDINITIIW NI HLONAT SAVM 009 fofohey oov cde o2e Thirtieth and Thirty-first Annual Mectings ian 40 The Kentucky Academy of Science Summary 1. A preliminary study has been made on the separation of certain of the ether-insoluble pigments found in tobacco. 2. By use of chromatographic adsorption analysis, five of these pig- ments have been separated. 3. Absorption spectrum curves have been determined for the five pig- ments thus separated. ; 4. The presence of still other of these pigments in tobacco has been indicated. References 1. Griffith, R. B. 1942. Master’s Thesis, University of Kentucky. . Strain, Harold H. 1942. “Chromatographic Adsorption Analysis.” Interscience Publishers, Inc., New York. §. Sheard, C. and States, M. H. 1941. J. Optical Soc. Am., 31: 64-69. 4. Verhagen, Dirk 1943. Master’s Thesis, University of Kentucky. nN Thirtieth and Thirty-first Annual Meetings 4] ONTOLOGY OF CONSCIOUSNESS* Lester S. O’ BANNON Experiment Station University of Kentucky There are two basic approaches to the ontology of consciousness. One leads through the physical categories, space, time, matter, motion and force. ‘Those who follow this path either deny the existence of consciousness as something to be reckoned with, or attempt to explain it as a natural consequence of physical activity. his method con- forms to the rule of parsimony, and for that reason is to be com- mended, but the results invariably are vague and unconvincing. The second approach is by way of postulating an additional cate- gory. The new category assumes various names, such as mind, spirit or soul, or consciousness itself. “Those who follow this method evade the ontological problem. Then there are methods that combine the two basic approaches. Theorists who follow the conventional physical categories sometimes end with an entirely new category, such as, for example, eventness. It is difficult to see what there is in a unidirectional series of physical events that can produce anything to be called consciousness. Brain quality, or a qualitative aspect of brain events, is another category sometimes proposed. To call something a quality is merely to declare that it is swch as it is—which is far from being a satisfactory explan- ation. The problem seems to be to define consciousness in terms of some of those things which exist that do not already embrace consciousness itself. The physiology and evolution of consciousness are not points at issue. ‘The ontological problem is to show where consciousness 1s among existents. I conceive the universe to be kinetic-atomic, in four or five dimen- sions, depending upon whether I think of a single level or of two or more levels of kinetic atomicity. My speculative attitude towards the world may be described further as that of a man who spends his whole life in the middle of a stream swimming against a swift current: he has never seen either shore and he knows not whence the stream comes nor whither it goes. Under such circumstances one’s speculations are limited to the discovery of relationships and tendencies within one’s immediate environment. Such relationships as are discoverable are essentially * Read at the Thirtieth Annual Meeting, April 24, 1943, Louisville, Kentucky. 42 The Kentucky Academy of Science spatial and reversible and conform necessarily to the principle of con- servation. ‘Tendencies, on the other hand, are irreversible and temporal. My categories therefore consist of all of those attributes that are descriptive of the so-called physical world, such as space, time, matter and motion, and in addition, two indispensable tendencies which in physical terms would be described as potentials. One of these ten- dencies is characteristic of the physical, the non-organic or non-living world, and is to be identified always with the second law of thermo- dynamics. The other tendency is characteristic of the biological world. That is, it is the distinguishing characteristic of those parcels of mat- ter that strive to maintain and build up their state of organization, as opposed to the tendency toward disorganization which all of nature otherwise possesses. “hese tendencies are as real as kinetic atomicity— no more transcendental, and no less empirical. We are able to recognize these two opposing tendencies only be- cause we are predominantly the one or the other. A log floating down- stream follows the tendency of the stream and does not have a sense of motion because it is itself a part of the stream. A man struggling against the current not only recognizes the opposing tendency but he also recognizes himself as being in some way different from the stream. It is a conscious experience; and it is such as it is only because of these two opposing tendencies. When Descartes said cogito, ergo sum, he no doubt made a mature and recondite observation, but he overlooked the crux of the onto- logical problem. The proper place to start is at the final o in cogito; that is, with the first person singular, present tense. The perimeter of the O is the periphery which separates the I from the not-I. This is obvious enough and identifies what most of us mean by conscious- ness, but it does not go far enough. In the field outside the perimeter an. arrow pointing downward should be placed to represent the ten- dency of the physical world. This tendency is an ontological fact cor- responding, perhaps, to the Aristotelian notion of becoming, except that in this case it should be called a going. Inside the perimeter one should place an arrow pointing upward to represent becoming in the positive sense; that is, a tendency to maintain and build up organ- ization. Consciousness, then, is to be identified with the vortex created by the two opposing currents. This setting of consciousness at the juncture of two opposing ten- dencies differs fundamentally from other points of view. Materialists possess only one tendency, that of the physical world, and hence are unable logically, and structurally, to account for either life or con- sciousness, even with the aid of all of the subtle and abstruse facts Thirtieth and Thirty-first Annual Meetings 4D of modern physics, physiology and physical psychology that may be assembled in the attempt to substantiate their argument. The biologically minded who consider consciousness as a product of evolution likewise limit themselves to a single tendency the origin of which remains obscure. In the end, it is not consciousness that the biologist achieves. Instead, he assumes the probability of conscious- ness and then proceeds to prove not consciousness itself but the cate- gory of consciousness. Dualists are those who assume a suitable category, and conse- quently, as we have already stated, evade the problem of consciousness. Dualists put one kind of stuff inside of the O and another kind out- side, but can find no structural connection between the two. The two kinds of stuff coexist and there is no ontological problem to be solved. Descriptively the dualist is correct, but his theory lacks both physical and logical coherence. I take the categories of space, time, matter and motion to be basic to both the living and the non-living. My dualism comes from recog- nizing two opposing tendencies that occur co-temporaneously in one stuff; whereas, in other dualistic systems the stuff itself is divided in accordance with the dictates of a three-dimensional logic. Thus, in addition to matter and indefinite or indifferent motion, I see positive and negative tendencies, or potentials, which are to be identified with additional dimensions. This is a consistent coherent system which embraces all that we as participants in the universal flux are capable of perceiving. There is no matter without motion and no motion with- out matter. These together constitute the single category, kinetic atomicity. But indifferent matter and motion is meaningless and senseless. ‘here must be determinateness and purposiveness, explicit pattern and design. These qualities can be achieved in nature if, and only if, nature follows tendencies that are eternal and absolute and not capricious. Furthermore, if there is one tendency, there also must be an opposing tendency. My contention is that generically and cate- gorically there are only two tendencies; that one opposes the other; that consciousness mediates these tendencies; and that consciousness is, ontologically, where these two tendencies meet. I do not consider consciousness as being intrinsic solely to brain events. Brain events are sub-conscious events that involve sub-generic or specific tendencies such as those intrinsic to the heart or liver. Con- sciousness is intrinsic to the organism as a whole and not to a particular organ. The brain is essential to consciousness only because it is the means by which the unity of the body is attained. Unity is attained when the body acts as a whole in relation to other bodies. But this does not mean that the body is one of several stones tumbling down 44 The Kentucky Academy of Science a hill. On the contrary, it means a stone come to life and possessing the ability to climb the hill. ‘There is an important distinction here that should not be over- looked. Imagine a stone climbing a hill. Can you imagine a stone climbing a hill merely by the changing of its internal configuration? In order for a stone to climb a hill it must reach out and grab some- thing to pull or push itself along. It must organize its internal re- sources for this purpose, and maintain those resources. Also, it must act as an integrated, coherent whole because the stone as such will not reach the top of the hill if it becomes disorganized and leaves part of itself behind. But in order for the stone to be able to reach out and grab success- fully it must possess the means of distinguishing both the presence (perception), and the probable utility of appropriate objects (concep- tion). And it will need more than sense receptors and more than motor effectors. It will need something to mediate between itself and the hill; something wherein strategic interpretations can be made, meanings evaluated, volitions formulated, and other distinctions de- lineated. ‘This is the function of consciousness. Consciousness 7s where the living stone meets the hill. It is at the conjunction of two opposing tendencies, one of which is the slope of the hill; the other, the will to climb the hill. I can see only two objections to this point of view. One is that organic and non-organic tendencies might be considered as pointed in the same direction but as differing relatively. But further thought will reveal that such relative tendencies when viewed effectively in terms of relative motion or relative change would require a third referent necessarily absolute. Such an absolute we do not have. It is tempting to name either space or time; but this is untenable since we know that space and time are not absolute. Space and time are only descriptive of kinetic atomicity. Having no absolute referent, we therefore cannot imagine organic and non-organic systems both mov- ing in the same direction with tendencies differing relatively in such a way as to account for differences between the respective character- istics of each system. ‘There is no absolute system of another sort by which we can account for the resistance or drag that may cause one tendency to lag behind the other. The two imagined parallel systems become in effect a single system with a single unidirectional tendency. As a matter of fact, we have no right to entertain such an imagin- ing as a single system with a single unidirectional tendency. If we were a part of that system, as logs floating downstream, we would not be able to recognize either the system, ourselves as individuals, or the directional tendency of the system. It is only because we are a system, Thirtieth and Thirty-first Annual Meetings 45 or part of a system, having a tendency paralleling an opposite direc- tion that we are capable of cognizing the other opposing posited direction. The question may arise: Since opposite tendencies are in a strict sense relative, where is the third absolute or referent for gauging the tendencies? The answer is that this absolute is at the conjunction of the two tendencies. When two bodies meet head-on in impact there is nothing more absolute as a referent for their respective previous directions of motion than the event of impact itself. This meeting of opposite tendencies coupled with something sub- stantial through which they operate constitutes for us our universe, including ourselves as representative of one tendency, and the objec- tive world as representative of the other. To thus conceive our uni- verse is consistent with our schedule of minimum data for a rational philosophy. These data are: (1) something substantial to yield ap- pearances; (2) change; and (3) direction of change. The second likely objection that I see to my point of view may be stated as follows: If you place consciousness at the conjunction of opposing tendencies, one of which represents the physical world and the other the biological world, would it not be just as logical to say the environment is conscious as to say the organism is conscious? We know from personal experience that the organism is conscious, but we do not know and cannot believe that the environment is conscious. This query illustrates the fallacy of mosaicism: the irrational doc- trine that the body can be separated and lifted bodily from the envir- onment, either logically or physically or both. It is an obvious fact that there is no division line between the body and the environment. The same kind of atoms are contained in each. The atoms of each are joined by the same kind of forces and possess the same degree of dimen- sional freedom. Contained in one inseparable stuff the two opposite tendencies constitute the only distinction that can be made between the living and the non-living. We have indicated in another place! that consciousness is char- acteristic of a particular level of kinetic atomicity. It is not essential that every concretion of physical stuff that shows signs of living shall be conscious. Since consciousness is a personal matter we cannot know for sure but we can be reasonably certain that any living body that possesses the means of organizing and directing its parts in such a manner as to participate in its environment as a single unit possesses also the capacity for being conscious, and that it is conscious when it exercises that capacity. 1“On the Five-dimensional Mechanism of Knowing,’ University of Kentucky Research Club Bulletin, December 1942. 46 The Kentucky Academy of Science The brain of course performs a strategic function in establishing the unity of the organism. In case of injury to the brain, in natural sleep, or in hypnosis, the brain may or may not be active in a psychical sense. It certainly would continue to function in both a physical and physiological sense. The essential thing that happens under those conditions is that the unity of the body is disrupted. The social unity is dissolved on the cellular level, exactly as social consciousness on the molar level would be destroyed if the means of communication be- tween individuals should become inoperative. Furthermore, I am not sure that we know where within the body the functional aspects of the brain begin or end. It seems to me that consciousness is intrinsic to the organism but not to a particular organ. Since the stuff of the organism cannot be separated either logically or physically from the stuff of the environment, and since we can hardly deny that conscious- ness is peripheral, then the only place where consciousness can exist is at the juncture between that tendency which the organism has to pre- serve itself and that opposite tendency which the stuff of the organism has to disintegrate. Thirtieth and Thirty-first Annual Meetings 47 INFLUENCE OF MANGANESE INTAKE UPON UREA EXCRETION*+ J. T. SKINNER and J. S. McHarcur Contribution from the Department of Chemistry of the Kentucky Agricultural Experiment Station In recent years, as Borsook and Dubnoff have pointed out in their review (/), there has been an accumulation of data which indicates that urea formation may not occur through the ornithine cycle as pro- posed by Krebs (2). Assuming the validity of Krebs’ theory, one would expect a reduction in urea output in manganese-deficient animals since arginase, an enzyme essential to the ornithine cycle, is markedly activated in vitro (3-6) by manganese. In their investigations con- cerning the nature of manganese deficiencies Shils and McCollum (7) and Wachtel et al (8) detected no differences between normal and manganese-deficient rats as regards nitrogen metabolism. No data on nitrogen excretion are included in either report. A thorough study of urea excretion, at different levels of nitrogen intake and on different rations, as affected by manganese intake should furnish information which would be of aid in evaluating Krebs’ theory (2) of urea formation. Procedure Animals with limited reserves of manganese were prepared by placing stock colony rats with their litters, when the latter were two weeks of age, in cages provided with wire false bottoms and feeding them a milk-iron-copper ration until the young weighed 40-50 gm. The litters were then divided so as to provide a litter-mate control of the same sex and approximately the same weight for each animal receiving a manganese supplement. Series I received a basal ration of whole milk supplemented with 1.0 mg. of iron and 0.1 mg. of copper per 100 cc. Analysis of composite samples of the milk representing several days’ deliveries showed it to contain 0.053 mg. of manganese per liter. ‘The second series of ani- mals were fed a solid ration having the following percentage composi- * The investigation reported in this paper is in connection with a project of the Kentucky Agricultural Experiment Station and is published by permission of the Director. + Read at the Thirty-first Annual Meeting, April 29, 1944, Lexington, Kentucky. 48 The Kentucky Academy of Science tion: crude casein 18, sucrose 75.7, salts! 5, fortified corn oil? 1, and a mixture® of B vitamins 0.3. Since it was planned to use the same ani- mals in another study in which a magnesium deficiency was desired, this ration was low in magnesium as well as in manganese. On the basis of solids the two rations contained 0.4 and 1.5 p.p.m. of man- ganese, respectively. Addition of the respective manganese supple- ments raised the levels of manganese to approximately 8 and 500 p.p.m. The paired feeding method was employed with both rations. As a ‘rule, food intake of each pair was limited by the amount consumed ad libitum by the rat on the control (low manganese) ration. One series of urines, however, was collected from animals on the solid ration when the food intake was limited approximately to one-half the average normal consumption. After the rats were maintained on their respective rations for about two months, urine samples from the individual animals were collected by placing their cages over funnels the stems of which extended into bottles containing a small amount of toluene. Bottles were changed daily, the urine samples being stored in the refrigerator until the end of the three-day collection period when the samples of a given rat were combined for analysis. Urea was determined by a modification of the Van Slyke and Cullen procedure (9). One series of urines was analyzed for ammonia content. This compound was determined, after addition of potassium carbonate to an aliquot of the urine, by aera- tion into N/20 sulfuric acid. Results When one compares the urea nitrogen output with nitrogen intake, as recorded in Table I, it will be observed that in neither the control nor the manganese groups was there a constant relationship between the two nitrogen values. In order to check on this inconsistency the urines in Series I were also analyzed for ammonia. ‘The data on am- monia excretion, Columns 6 and 7, show that for each pair the low animal as regards urea excretion invariably excreted the greater amount of ammonia. For the control group the combined urea and ammonia outputs accounted for an average of 74 per cent of the nitro- gen intake as compared with 84 per cent for the animals receiving the manganese supplement. In this series four of the six control animals 1Except for omission of magnesium and manganese compounds this salt mix- ture was essentially that of Phillips and Hart, J. Biol. Chem. 109, 657 (1935). *Carotene 0.133 gm., viosterol 1 gm., alpha-tocopherol 0.4 gm., 2-methyl-1, 4-naphthoquinone 10 mg., corn oil 100 gm. *'Thiamine hydrochloride 20 mg., pyridoxin hydrochloride 20 mg., riboflavin 50 mg., calcium pantothenate 0.1 gm., nicotinic acid 5 gm., p-aminobenzoic acid 5 gm., inositol 10 gm., choline chloride 10 gm. Thirtieth and Thirty-first Annual Meetings 49 excreted more urea than did their mates which ingested twenty times as much manganese, but the situation was reversed in Series II on a normal food intake. When the nitrogen consumption of the animals in Series II was limited to 0.408 gm. per rat, approximately one-half the normal intake, the urea nitrogen excreted by the controls aver- aged 0.3156 gm. as compared with 0.2937 gm. for the animals receiving additional manganese to the extent of 500 p.p.m. of ration. Only one rat in the manganese group excreted more urea than its control mate when food intake was restricted. This fact is of interest in view of the tendency of the same animals to excrete more urea than the con- trols when the protein intake was normal. Whether the manganese supplement enabled the animals to utilize a suboptimal amount of protein more efficiently than did the controls cannot be determined from the data available. In Column 5 the urea excretion of each man- ganese rat is expressed as percentage of the output of its respective control mate. The percentages range from 82 for Pair 5 on the milk ration to 139 for Pair 10 when consuming a normal amount of the solid ration. Considering the data for the three series one finds that in 7 instances the manganese animals excreted more urea than their TasLeE I. Nitrogen Excretion on High and Low Manganese Intakes Urea-N output NH3-N output Pair : N No. Intake Control Mn-fed Mn-fed/ Control Mn-fed Control Series I (Milk-iron-copper ration) gm. gm. gm. per cent gm. gm. 0.6875 0.4302 0.3785 88 0.0235 0.2049 0.6050 0.3872 0.3397 88 0.0539 0.2298 0.7563 0.3852 0.3329 86 0.0459 0.1905 0.5511 0.2981 0.3353 113 0.1262 0.0961 0.5225 0.3232 0.2658 82 0.0436 0.1464 0.8250 0.3758 0.4404 117 0.4036 0.3495 0.6579 0.3666 0.3488 95 0.1161 0.2029 (Solid ration) Normal intake eis 0.4408 0.4281 97 ee 0.4116 0.4721 115 9* 0.5516 0.4619 84 10* 0.3788 0.5256 139 11* 0.4431 0.4970 112 12* 0.4952 0.5123 104 Av. 0.4535 0.4828 106 Series II (Solid ration) Reduced intake 0.3423 0.3039 89 0.3060 0.3242 106 0.3201 0.2955 92 0.3025 0.2910 96 0.3079 0.2790 91 0.3147 0.2688 85 0.3156 0.2937 93 * Average of two periods. ** Average of three periods. 50 The Kentucky Academy of Science respective controls. he average urea excretion of all manganese groups amounted to 101 per cent of the average output of the con- trol groups. Summary Urea excretion was determined on rats at varying levels of food con- sumption and on low, medium and high intakes of manganese. In the manganese groups the urea excretion ranged from 82 to 139 and averaged 101 per cent of that excreted by the respective pair-mate controls. In 7 out of 18 comparisons it was observed that the manganese-fed rat excreted more urea than its pair mate on a low-manganese ration. Bibliography . Borsook, H., and Dubnoff, J. W., Ann. Rev. Biochem. 12, 194 (1943). . Krebs, H. A., and Henseleit, K., Z. physiol. Chem. 210, 33 (1932). . Edlbacher, S., and Pinosch, H., Z. physiol. Chem. 250, 241 (1937). Edlbacher, S., and Baur, H., Z. physiol. Chem. 254, 275 (1938). Edlbacher, S., and Baur, H., Naturwissenschaften 26, 268 (1938). . Richards, M. M., and Hellerman, L., J. Biol. Chem. 134, 237 (1940). . Shils, M. E., and McCollum, E. V., J. Nutrition 26, 1 (1943). . Wachtel, L. W., Elvehjem, C. A., and Hart, E. B., Am. J. Physiol. 140, 72 (1943). . Van Slyke, D. D., and Cullen, G. E., J. Biol. Chem., 19, 211 (1914). Oo cont oO FOO Ne Thirtieth and Thirty-first Annual Meetings 5] DERIVATIVES OF 1-DIMETHYLAMINOMETHYL- 2-NAPHTHOL* A Progress Report GERALD F. GRILLOT Department of Chemistry University of Kentucky The purpose of this investigation has been the preparation and the detection of any local anesthetic action of the derivatives of 1-dimeth- ylaminomethyl-2-naphthol (Compound VIII: NR, = N(CHs3)»s. See plate, page 55). Since 1-dimethylaminomethyl-2-naphthol as well as many com- pounds whose derivatives show local anesthetic power are synthesized by the Mannich reaction, some information concerning this reaction follows: Although ‘Tollens (/, 2) observed condensations of this type in 1903, and Auwer and Dombrowski (3, 10) reported the condensa- tion of phenols with formaldehyde and secondary amines in 1906, no extensive study of this reaction was begun until about 1917 (4, 5). In this year Mannich began studies on this reaction which now bears his name. The Mannich reaction is a condensation of an ammonium salt or an amine with formaldehyde and an organic compound which con- tains a fairly active hydrogen atom. Secondary amines and their salts usually produce a single product, while primary amines, their salts and ammonium salts lead to a mixture of products which usually are difficult to separate. Among the compounds which contain fairly active hydrogen atoms and which take part in this condensation are ketones, aldehydes, and phenols. If ethyl methyl ketone (Compd. I) is condensed with formaldehyde and dimethylamine, 3-methyl-4-dimethylamino-2-butanone (Compd. Ia) is produced. ‘This reaction is an example of the Mannich condensa- tion. ‘This aminoketone can be reduced to the corresponding amino- alcohol, 3-methyl-4-dimethylamino-2-butanol (Compd. II). If this al- cohol is treated with p-nitrobenzoyl chloride, the hydrochloride of the p-nitrobenzoyl ester of this alcohol (Compd. HI: R = p-NO.C;Hy CO-—) is obtained, and this ester on reduction produces “Tutocaine” (Compd. III: R = p-NH,C,H,CO—). “Tutocaine” produces good sur- face anesthesia, as well as anesthesia by subcutaneous injection. An example of an aldehyde used in the Mannich reaction is iso- butyraldehyde (6). When it is condensed with diethylamine and for- maldehyde, 3-diethylamino-2, 2-dimethylpropionaldehyde (Compd. IV) * Read at the Thirty-first Annual Meeting, April 29, 1944, Lexington, Kentucky. De The Kentucky Academy of Science is obtained. This can be reduced to an alcohol, (Compd. V) which is much more stable than the aldehyde. ‘The p-aminobenzoyl ester of this alcohol (Compd. VI: R = p-NH,C,H,CO-—) possesses both sur- face and subcutaneous anesthesia and has been used as a substitute for Cocaine under the name of “Larocaine.” One of the simplest examples (7) of the Mannich reaction using a phenol as the component with an active hydrogen atom, is. the con- densation of phenol with formaldehyde and diethylamine, whereupon 2-diethylaminomethylphenol (Compd. VII) is produced. Auwer and Dombroski (/) condensed beta-naphthol with formaldehyde and piper- idine and obtained 1-piperidinomethyl-2-naphthol [Compd. VIII: NR», = N(CH,);] in good yields. Grillot and Shriner (8) prepared the hydrochloride of the p-aminobenzoyl ester [Compd. IX: NRs= N(CH));; R’ = p-NH.,C,H,CO—] of this compound. It was slightly more toxic than Novocaine, and produced good surface anesthesia. Another compound prepared by Auwer and Dombroski by the condensation of beta-naphthol with formaldehyde and dimethyla- mine (J, 10) was dimethylamonimethyl-l-naphthol (Compd. VIII: R = CHs3). The author of this paper has prepared this compound, and a method for its synthesis is discussed in the experimental part. It was obtained as colorless plates that melted at 74-74.5°C. This substituted naphthol was treated with benzoyl chloride, and the hydrochloride of the benzoyl ester (Compd. IX: R = CH;; R’ = Cg;H;CO—) was ob- tained as a colorless amorphous powder which decomposed at 174- 175°C. The hydrochloride of the p-nitrobenzoyl ester (Compd. IX: R = CH; R’ = p-NO.C,H,CO—) was obtained as pale yellow needles by treating the naphthol base dissolved in toluene with p-nitrobenzoyl chloride. ‘The p-nitrobenzoy] ester was dissolved in alcohol and was hydrogenated, using a platinum catalyst; the hydrogen was introduced at room temperature and about 2-3 atmospheres pressure. Although approximately the calculated amount of hydrogen was absorbed, no crystalline p-aminobenzoyl ester (Compd. IX: R=CH3; R’=p-NH»2— C,;H,CO-—) could be isolated from the solution. An attempt was also made to prepare the cinnamoyl ester (Compd. IX: R = CH3; R’ = C;H;CH = CHCO—) of dimethylaminomethyl-2-naphthol, but the viscous mass resulting from the action of cinnamoyl chloride on this substituted naphthol could not be purified. The anesthetic properties of the p-aminobenzoyl esters of the com- pounds derived from the Mannich reaction can be suspected because of their resemblance to the structure of “Novocaine.”’ This is evident when the structure of “Larocaine”’ (Compd. VI: R = p-NH,C,H CO—) and the p-aminobenzoyl ester of 1-dimethylaminomethyl-2- naphthol (Compd. IX: R = CH;; R’ = p-NH.C;H,CO—) are com- Thirtieth and Thirty-first Annual Meetings 53 pared to “Novocaine” (Compd. X: R= p-NH,C,H,CO—). (Note similarity of the enclosed portions of these three molecules as shown on the included plate, page 55.) Since piperidine (K, — 1.6 x 10-*) is a stronger base than dimeth- ylamine, (K, = 7.4.x 10-4), the dimethylaminomethylnaphthol would be expected to be a weaker base than the piperdinomethylnaphthol. The hydrochloride of the weaker base will be more readily hydrolyzed to the free base, and will be absorbed into the nerve tissue more readily than the stronger base. ‘Then the derivatives of the weaker base should produce greater anesthetic action, but at the same time it should be more toxic (9, //). ‘Therefore one should expect that 1-dimethylamino- methyl-2-naphthol derivatives should be slightly more powerful as anesthetics than the derivatives of the corresponding piperidine deriva- tives, but at the same time should be slightly more toxic. After the synthesis of the 1l-dimethylaminomethyl-2-naphthol derivatives they will be tested for local anesthetic action, and then it should be pos- sible to compare them qualitatively with the corresponding piperidine derivatives in order to see if the above predictions are supported by the evidence. Experimental 1-Dimethylaminomethyl-2-naphthol (3): To 29 g. of @Q-mnaphthol dissolved in 150 ml. of alcohol (95%) was added 27 g. of 33% dimeth- ylamine solution. The resulting solution was cooled and 15 g. of formalin (35-40°% formaldehyde solution) was added slowly with stir- ring. When this was seeded with crystals and was permitted to stand a discolored crystalline mass formed which was filtered and dried. ‘The yield of this crude product amounted to 28 g. or 70% of the theoretical. The melting point of this product was 73-74°C. This product was recrystalized from alcohol using charcoal as a decolorizing agent. Beau- tiful colorless plates were obtained which melted at 74-74.5°C. 1-Dimethylaminomethyl-2-naphthyl Benzoate Hydrochloride: ‘To 8 g. of 1-dimethylaminomethyl-2-naphthol dissolved in 100 ml. of cold dry toluene was added slowly a solution of 6 g. of benzoyl chloride also dissolved in 100 ml. of cold dry toluene. ‘The ester immediately pre- cipitated. ‘The reaction mixture was permitted to stand over night. It was then filtered and washed with toluene, then ether. It was then placed in a desiccator for several days. Decomposition point of this material was 174-175°C. ‘The yield amounted to 12.5 g. which was 89% of the theory. Anal. caled. for C,,H,,0,N Cl: N,4.10; Cl, 10.3. Found: N, 4.09, 4.03; Cl, 9.97, 10.0. 54 The Kentucky Academy of Science 1-Dimethylaminomethyl-2-naphthyl p-Nitrobenzoate Hydrochlo- ride: Twelve grams of |-dimethylaminomethyl-2-naphthol was dis- solved in 100 ml. of dry toluene and this solution was added to 11.5 g. of p-nitrobenzoyl chloride which was also dissolved in 100 ml. of dry toluene. The resulting material was heated on a hot water bath for three hours and was then permitted to stand overnight. ‘he residue was filtered and dried. A pale yellow powder was obtained in a nearly quantitative yield. It decomposed at 184-185°C. When it was recrys- talized from alcohol (95%), pale yellow needles were obtained. Anal. calcd. for Cy>H,gO,NoCl: N, 7.27; Cl, 9:2. Found: N> 742; (Ass (Cll, S805 nels). Attempted preparation of 1-Dimethylaminomethyl-2-naphthyl p-Aminobenzoate Hydrochloride: Ten grams of |-dimethylamino- methyl-2-naphthyl p-nitrobenzoate hydrochloride was suspended in 250 ml. of alcohol (95%) and 0.2 g. of Adam platinum catalyst (/2) was added. This solution was then shaken with hydrogen at 2-3 atmospheres pressure and at room temperature; 0.082 moles of hydro- gen were absorbed. (0.078 moles of hydrogen is the theoretical quan- tity that should have been absorbed.) The platinum residues were removed by filtration, the resulting clear solution was evaporated to 50 cc., and was then placed in an ice chest. No crystallization occurred over an extended period of time. Cinnamoyl Chloride (/3): After the author failed to obtain any cinnamoyl chloride by the action of phosphorus pentachloride on a half mole of cinnamic acid, he devised the following procedure for the preparation of this acid chloride: Twenty-eight grams of commercial thionyl chloride (10% excess) was added in small portions to 30 g. of cinnamic acid (C.P.). When all the thionyl chloride was added the reaction mixture was warmed to about 50°C. until the vigorous reaction that first occurred nearly subsided. The residue contained little unreacted thionyl chloride and distilled as a pale yellow oil at 128-131°C. at 14-16 mm. of pressure. On standing, the distillate set to a crystalline mass. ‘The yield amounted to 28.4 g. or 849%, of the theory. Attempted preparation of 1-Dimethylaminomethyl-2-naphthyl Cinnamate Hydrochloride: To 6.5 g. of cinnamyl! chloride dissolved in 75 ml. of dry toluene was added 8 g. of 1-dimethylaminomethyl-2- naphthol which was also dissolved in 75 ml. of dry toluene. The re- sulting reaction mixture was permitted to stand overnight and a gummy residue formed. Attempts to recrystallize this mass were unsuccessful. Thirtieth and Thirty-first Annual Meetings 55 CH,COCH+CHO+(CH,), NH —> CH,COCHCH,N (CH, ) => GH. Cre if Ta CH,CHOHCHCH,N (CH,), > CH.CHEHCH.NCH),HCL CAL OR CH, al mm CH, CH, GH), NCHECHO- (GHNCHECHOH+ quel “tf CH, CH, Ww Frye’ ua O CH,N(C2Hs)2 AVAOE CH.NR, | CH,NRZHCL COM Coe CO AVAILE IX | CH.NICH.).“HCL | CH,[OR] x Bibliography . Van Merle and Tollens, Ber., 36, 1351, (1903). Shafer and Tollens, Ber., 39, 2181, (1906). . Auwer and Dombrowski, Ann., 344, 284, (1906). Mannich and Kroesche, Arch. Pharm., 250, 647-67, (1912). . Mannich and Kathers, Arch. Pharm., 257, 18, (1919). . Mannich, Lesser and Selten, Ber., 65, 380, (1932). Gormley, Wm. T. Jr., Master’s thesis, University of Kentucky, 1945. . Grillot, G. F., Doctor’s thesis, University of Illinois, 1940. Goodman and Gilman, The Pharmacological Basis of Therapeutics, page 242, Macmillan Co., (1941). 10. German Patent to Friedr. Baeyer and Co., D.R.P. No. 89, 979. 11. Nisbet, J. Chem. Soc., 1237, (1938). 12. “Organic Synthesis,” Collective Vol. I, p. 452, John Wiley & Sons, N. Y. (1932). 13. Claisen, Antweiler, Ber., 13, 2124; Meyer, Monat., 22, 428. Per ARON > 56 The Kentucky Academy of Science INFLUENCE OF POPULATION NUMBER ON EGG PRO- DUCTION IN THE FOUR-SPOTTED PEA BEETLE, BRUCHUS QUADRIMACULATUS FABR.* ALFRED BRAUER Department of Zoology University of Kentucky Introduction ~ A study by Park (1932) on the relationship of numbers to initial population growth in the flour beetle, Tribolium confusuwm, showed that population growth is more rapid in this insect when there is an intermediate degree of crowding, than when there is a larger or smaller number of individuals present in the initial group. He later identified (Park 1933) two opposing ecological factors which favor initial populations of intermediate numbers. The first of these is recopulation, a stimulus to egg laying, favored when the population is large. The second, cannibalism, prevails in a given quantity of meal overcrowded with eggs and with feeding larvae. A similar study by Maclagen (1932) on Drosophila, showed that the reproductive rate is unaffected by number of pairs originally present, or through a number of degrees of crowding within the cul- ture medium. Several years ago in our laboratory it was observed that in the beetle Bruchus quadrimaculatus repeated copulation appeared to stimulate egg laying. Thinking that this condition would be favored in a given volume if the number of individuals were increased, the problem was placed on an experimental basis with stock beetles then available. ‘The result was that single pairs always yielded a greater number of eggs per female than did the group cultures. These preliminary observations warranted further experimenta- tion, and the present paper is concerned with more exacting tests relative to the influence of population numbers on egg production in this beetle. Methods Cultures of bruchids were started from wild stock; the first and successive generations of emergents were used for experimentation. Sterilized black-eyed peas provided the nutrient material. Cultures were maintained in shell vials 314” x 1”, and varying degrees of crowding were obtained by confining one, two, four, or six pairs of * Submitted May 1, 1945. Thirtieth and Thirty-first Annual Meetings 57 beetles within a vial containing twenty to twenty-four peas and plugged with cotton down to the level of the peas. The surface of the cotton in contact with the peas was covered with a layer of black paper so that eggs which otherwise would have been lost in the cotton could be seen and included in the count. ‘The volume occupied by the peas was 15 cc., and this was considered as the unit, or standard volume. ‘This volume of peas was doubled in two of the experiments. Bruchids are sexually dimorphic and readily distinguishable. For experimentation they were separated from brood cultures at the time of their emergence from egg-infested peas, or within an hour there- after. The first eggs of newly emerged bruchids are not usually de- posited by them until about six to ten hours after their emergence. Eggs were counted daily in the group cultures, and either daily or on alternate days in I-pair cultures until the death of the parent beetles. At the final count, those eggs deposited on the sides of the vial and on the black paper over the cotton plug were added to those obtained on the peas. Counting errors were insignificant. The first series of experiments consisting of |-pair, 2-pair, 4-pair, and 6-pair cultures were run simultaneously with first generation emergents of the wild stock. Here one series of cultures served as control to each of the others. For each series run thereafter with emergents of later generations, a l-pair control series was run simul- taneously with emergents of the same culture. Uniform conditions of incubation and handling were maintained and observed. Experimental 1. INFLUENCE OF VARYING DEGREES OF CROWDING ON EGG PRODUCTION Table | summarizes records of one to six pairs of beetles confined on the standard volume of 15 cc. of peas. This shows an almost unt- form and steady decline from 94 + 2.8 eggs per female for single male-female pairs down to 63 + 2.2 eggs per female in 6-pair cultures. The decline between any two successive series is not significant sta- tistically, but becomes so between alternate successive series. In order to determine whether or not the difference in egg produc- tion between single pairs and 6-pair groups would be maintained through several generations, a test was made with seventh generation emergents. By chance the brood culture selected to supply the test beetles contained an especially fecund stock, and egg production was considerably greater than it was in the previous test (Table 1, a and b). Average production in the l-pair series was 102 + 5.7, despite the fact that one female produced only 61 eggs. Six of the ten females in the series produced over 100 eggs cach. (Fig. 1) Average production in the 6-pair cultures was increased by 15 eggs 58 The Kentucky Academy of Science per female over that of the equivalent group in the first test. Never- theless the average production per female remained below that of the control series by about the same percentage as it had in the first test. Rate of egg deposition usually reached its maximum during the second 24-hour period, and thereafter declined steadily to the end of the adult life span of eight to ten days. The lower egg production of the first day is undoubtedly due to the fact that eggs are seldom de- posited prior to six hours after fecundation of virgin females. Taste |. Influence of varying degrees of crowding on egg production in bruchid cultures maintained at standard volume of 15 cc. (a) Number of No. of Max’m no. Min’m no. Av. no. Standard pairs per enltnes eggs per eggs per eggs per devia- culture female female female tion hn 20 115 714 94.0+ 2.8 12.4 aie See 10 113 67 86.6 = 3.7 12.5 A * a w 7 88 72 79.0 + 2.3 7.7 Ge eee 10 73 54 63.0 + 2.2 10) (b) Supplementary tests involving a particularly prolific strain. eee Setar 10 124 61 102.0 + 5.7 19.0 G2 eee eee 10 93 62 78.5 + 3.4 11.3 Average in each case is followed by its standard error. Note that under the conditions of the experiment, progressive crowding of beetles was accompanied by declining egg pro- duction. Tasie 2. Interrelationship between number of pairs and volume, and ege production in bruchids. No. of Volume No. of Max’m no. Min’m no. Ay. per Standard pairs per of culture cultures eggs per eggs per female devia- culture in cu. cm. female female tion cme etepre tee i 15 20 115 74 94.0 + 2.8 12.3 Dy pteeas So ee, 30 10 138 54 93.5 + 7.6 25.2 Di ee ON 15 10 113 67 86.6 + 3.7 12.5 Pd apes dee CEL 30 10 121 72 88.4 + 4.7 15.6 Note that in cultures containing only one or two pairs of beetles only minor changes in egg production occurred when culture volume was doubled, probably because of minor changes in ecological factors involved at those degrees: of crowding. Doubling number of pairs when volume was held constant depressed egg production because of crowding. 2. INTER-RELATIONSHIP BETWEEN NUMBERS OF INDIVIDUALS AND CUL- TURE VOLUME, AND EGG PRODUCTION Several tests were made in which the culture volume was increased to two times standard volume, or to 30 cc., in I-pair, and in 2-pair series. The l-pair series of Table l-a served as a general control (Table 2). In this test the l-pair series on double volume, gave an average of 93.5 + 7.6 eggs per female; remarkably near that of the control Thirtieth and Thirty-first Annual Meetings 59 series. Here however, the standard deviation, hence the standard error, was increased x 2, over that of the control, and over that of any other 1]-pair series. Increasing the standard volume x 2, for a 2-pair series, increased the average production per female somewhat, or to a point between that of the l-pair and the 2-pair series. Although not statistically sig- nificant, this is in agreement with preceding tests and with expected results. 3. INFLUENCE OF FEGUNDATION ON EGG-PRODUCTION IN BRUCHID CULTURES In Bruchus, at least one copulation is required to initiate oviposi- tion, but during the productive period it occurs repeatedly. In order to test the importance of repeated copulation on the production rate, and the number of eggs deposited, two tests were made with 8th gen- eration emergents. In the first of the tests (Table 3, a) females were paired with males for about six hours. Each pair was observed to have copulated at least once, but it is probable that this occurred several times. The females were then separated from the males and isolated singly in unit-volume vials of peas. In only one female was the average number near that of the male-female control series. The rate of deposition in all but the one mentioned above declined sharply after the third day, thus showing that repeated copulation exercises some control over egg laying. Tasie 3. Influence of repeated copulation on egg production in bruchid cultures maintained at standard volume of 15 cc. (a) Treatment No. of Max’mno. Min’m no. Av. no. Standard cultures eggs per eggs per eggs per devia- female female female tion Females isolated singly after exposure to males for 6 hours ____.___ 10 83 36 59.0 + 4.4 14.8 Control. Male-female pairs not separted __.. 10 94 70 1923) == 228 8.6 (b) Females isolated by twos after exposure to males for 6 hours -_-_ 10 90 42 71.0 + 4.6 14.0 Control. Male-female pairs not separted —_ 10 105 71 90.0 + 3.7 12.0 Paired results in each case clearly indicate that re-copulation exercises a positive influ- ence on egg production in Bruchus. 60 The Kentucky Academy of Science Finally, to determine if contact with other females plays a part in ege production, a series was tested in which fecundated females were separated from males after six hours, and placed in pairs (female- female) within unit-volume vials of peas. Beetles for this test (Table 3, b) were from a different culture than those of the preceding test and hence required a separate control; however, the average number of eggs per female was lower than the number per female in the con- trol series. ‘The result of the test is consistent with the experimental results of each of the other tests. Discussion — Conclusion ‘These experiments show that maximum egg production for Bru- chus is realized when single pairs are confined in relatively small vol- umes of peas. As the number of beetle pairs is increased there is a steady decline for each additional pair. While the decline from 1-pair to 2-pair cultures is not sufficient to be statistically significant for the numbers tried, the decline becomes significant between l-pair and 4-pair cultures, or even between 2-pair and 4-pair cultures. If the curve and the abscissa (Fig. 1) were extended, they would meet at approximately 15. In other words, rate of decline remaining con- stant, egg production would reach zero in a concentration of 15 pairs in 15 cc. of peas. Undoubtedly other factors would alter the rate of decline however. O io) iG ro) 4 2 S A 25 6 Number of bruchid pairs per culture Average number eggs per ~] S) Ficure 1. Influence of varying degrees of crowding on egg production in bru- chid cultures of 15 cc. volume. Dotted curve is obtained from egg counts of Table Ib. Solid line curve obtained from Table Ia. Thirtieth and Thirty-first Annual Meetings 61 Within the limits of the tests, doubling the volume occupied by ]-pair beetles had no effect on the total number of eggs deposited, but resulted in a greater deviation from the average, the possible reason for which will be pointed out. Two-pairs in unit volume produced a noticeable if not significant decline in production, while 2-pairs in double volume produced somewhat fewer than 1-pairs, but more than 2-pairs in single volume. Repeated copulation as shown in ‘Table 3, is definitely a stimulus to egg laying, and probably requires no special comment; neverthe- less, sexual contact definitely appears to be the chief biotic factor re- sponsible for the drop in egg production of the more crowded cultures, namely: in four- and six-pair combinations a condition prevails which not only insures fecundation of each female, but increases the sexual contacts as well as contacts by moving individuals of both sexes. This does not afford females a sufficient interval to prepare for oviposition, for in order to do this the female first moves about, stops to massage the abdomen with metathoracic legs, then moves on to repeat the performance. When finally an egg is ready for release, some time is again required. ‘he behavior is repeated for each egg released. The laying behavior insures a distribution of eggs over numerous peas, a condition more favorable for larval feeding. Crowded cultures pre- vent the freedom required by females for this behavior in that they are continually aggravated by moving beetles or by soliciting males. When the culture volume is increased, some beetle pairs may make a sufficient number of sexual contacts to insure maximum production, while others do not, because there is greater opportunity for isolation; hence, the greater deviation from the average in these cultures. It is possible that 30 cc. is the critical maximum volume at which egg pro- duction will remain at maximum. This paper has only an indirect bearing on the problem of rate of population increase, as studied by Park on the meal beetle, Tri- bolium confusum (1932,1933), and by Maclagen (1932) for Droso- phila, namely in as far as the rate of population increase is affected by the total number of eggs lain per female. In this connection, however, another inhibiting factor has become apparent in this work and in laboratory observations on mass cultures of beetles on a limited number of peas. When peas become over-infested with eggs, the feed- ing larvae later crowd one another from their excavations in the peas before feeding is completed and maturity is reached. ‘These fail to pupate and mature, thus lowering the percentage of adult emergents under the number of eggs originally present on the peas. A priori reasoning would lead to the conclusion that for a given volume of peas there must be an optimum population number which 62 The Kentucky Academy of Science is large enough to insure sufficient sexual contacts between beetles for maximum production, yet not great enough to overstock the peas with eggs and larvae. Literature Cited Allee, W. C., 1938. Social Life of Animals. W. W. Norton & Co., Inc., New York. Maclagen, D. S., 1932. The Effect of Population Density upon Rate of Repro- duction with Special Reference to Insects. Proc. Roy. Soc. Br., 111:437-454. Park, Thomas, 1932. Studies in population physiology: The relation of num- bers to initial population growth in the flour beetle, Tribolium confusum Duval. Ecol., 13:172-182. Park, Thomas, 1933. Studies in population physiology: II. Factors regulating initial growth of Tribolium confusum populations. J. Exp. Zool., 65:17-42. Thirtieth and Thirty-first Annual Meetings 63 ABSTRACTS A preliminary report on the lichen flora of Kentucky.* Mary James ALLEN and Bruce E. Rawiines, University of Kentucky.—In the spring of 1941 the authors began systematic collections of the lichen flora of Kentucky with special reference to their habitats. Of the six hundred specimens collected, two hundred have been identi- fid and the identifications verified by G. G. Nearing. Collections have been made in Rockcastle,, Madison, Russell, Caldwell, Fayette, Mc- Creary, Pulaski, Whitley, Knox, Union, Jessamine, Wolfe, Menifee, Bath, and Rowan Counties. Interesting lichens discovered include Opegrapha cinera, collected in Whitley County, previously reported only from Florida by Fink; and Sticta pulmonaria, a northern species which we have found ex- tending to the southern boundary of the state. Probably the two most common species are Parmelia caperata, found on nearly every tree and rock, and Cladonia cristatella, the British soldier lichen, growing on soil in moist shaded areas. Teloschistes lychneus, a common yel- lowish lichen, has been collected in Jessamine County in the fruiting state which is unusual. A species of Verrucaria has been collected which appears to be new to science. Coccocarpia incisa, the rare sha- dow lichen, so named by reason of its dense, dark, webby hypothallus which grows its own shadow, has been found in abundance in Mc- Creary County. Dr. Bruce Fink is the only recognized lichenologist who has sys- tematically worked on the lichen flora of Kentucky, chiefly in Rock- castle County. We have found numerous species that he did not record from the state. The fact that very little reliable detailed work has been accomplished in this state makes it the undeveloped happy hunting ground for the lichenologist. ‘To prepare a comprehensive catalogue of the lichens of Kentucky is a project which will require many years. This paper is but a preliminary report and it is our hope that the work will be continued by others until completed. A new plant for the western hemisphere found in Kentucky.* Hot- Lis J. Rocrers, University of Kentucky.—A plant new to North Amer- ica belonging to the Labiatae was discovered by the author in Mc- Creary County. It has been identified by F. R. Fosberg of the United States National Herbarium as Mosla dianthera (Buck ex Roxb.) Max. He states that the former range of the plant was through southern and eastern Asia. In Kentucky Mosla has been found to be a weedy plant that pre- fers the low, sandy soil in the woods and in clearings along the river * Read at the 29th Annual Meeting, April 11, 1942, Lexington, Kentucky. 64 The Kentucky Academy of Science bottoms of the South Fork of the Cumberland River in McCreary and Pulaski Counties. Its known range extends from Worley in McCreary County over a distance of 40 miles of the course of the river in Pulaski County, where the south fork joins the Cumberland River. A thor- ough examination of the banks of the Cumberland River at Burks- ville failed to reveal any of the plants. No information is available as to whether Worley is the highest point on the South Fork River where the plant is established. It is quite probable that Mosla extends across the state line to some place in Tennessee. At only one place in this area have Mosla dianthera been found at a distance from the river, which means that the water to be backed up by the Wolf Creek Dam now under construction will cover at least 90 per cent of the plants. This location was found at Yammacraw where the species was quite abundant to a point about 800 feet above the river. Fosberg has published an account of the occurence of Mosla in Kentucky in the American Midland Naturalist for January, 1942. Aquatic plants of Jefferson County, Kentucky, and environs*. Mary SERGEANT, Jacobs School, Louisville, Kentucky.—The writer collected and made a study of aquatic plants for an interval of three years. This study includes the taxonomy, morphology and ecology of the aquatic seed plants of Jefferson County, Kentucky. Jefferson County is especially interesting for such a study because of the formerly large number of swamps and ponds, some of which have now been drained. The ponds of this county are of three types, permanent, tempor- ary and artificial. Each type of pond has a distinctive flora. From microscopic studies and observations of living plants, the writer concludes that submerged aquatics show the greatest modifi- cations. The presence of a highly developed system of intercellular spaces, reduced vascular system, very slight lignification and lack of cutin are the most distinctive features of true aquatic plants. Dis- tinctive features of emergent and subaquatic plants are the presence of diaphragms which cross the large internal air spaces and the pro- nounced variation in the growth of the leaves according to the watet content in which the lar is growing. The writer has collected and studied over one inaded species of aquatic plants, three of which were new records for Kentucky and one at least that once was abundant has all but disappeared due to drainage conditions. * Read at the 30th Annual Meeting, April 24, 1943, Louisville, Kentucky. INDEX = Volume 11 Abstracts, Volume 11, No..2 ....... es) TDI ate 8 Se A a LINO Ey a2 0X6 Ie a Ree ee ALL UN ACE AC A Allen, Mary Jane. A preliminary report on the lichen flora OH MCCHOUCkKy mM aOStract) (Pama « UES aA ce «dhe ee ree MEM othe Ltd Allen, W. R., Bottom-preferences of fishes in northeastern Ken turclayiStRea mash 5. 6/4 Fee RCP case ines eas OMe IME tae tar i leo Brauer, Alfred, Influence of population number on egg production in the four-spotted pea beetle, Bruchus quadrimaculatus fabr Clark, Minor E., Bottom-preferences of fishes in northeastern RemnuicksyyrStreanns) Nua iis ahs SUN a ae Ny A Ny a Rea ta yes Crouch, H. B., Localization and cleavage of cytoplasm in GELKAMG PATASIAG | PNOLOZO (1: oi. SU a Wat. ebay papa ene ela aa By te Davies,\P. AY Floral glands in Ailanthus altussima ....0)0.00- 2%. 4: The Root System of Ailanthus altissima................... Diachun, Stephen, Colonies of Bacterium tabacum on roots of wheat EMC MISE VETAN | CDASSES ME uf SNM all) so bland HOt azarae aidan ATED Age Grillot, Gerald F., Derivatives of 1-dimethylaminomethyl- ZHAI COOL) (aKOSTESS SLEPOONM 7. owe sae caste ii) Mae aay tal aseele ahs Hall, E. K., Spinal ganglion hypoplasia after limb amputation Ae e whet allt raitye (ADSiRACE) Main uNa tose Ue lnlrs cla ini) auieN recite a Alora Johnson, E. M., Colonies of Bacterium tabacum on roots of Wahler TaN Gy Se Vela MASSES pic ds ELE Ci) un aes C RMT) Alok aia Rreld)/straims ot tobacco mosaic’ (abstract) ))). 3...) 2 es McHargue, J. S., Excretion of manganese by nthe; rates. c(t eye clea Influence of manganese intake upon urea excretion........ OBannonmsWester'S.) Ontolory of COMSCIOUSIESS |). 1)0)0).)in)-\eiais es ee Pennebaker, G. B., The development of the uropygial gland in pigeons Rawlings, Bruce., A preliminary report on the lichen flora Ci MeneUcky y (albsiracty), meivamm se hi ciao eisie creda, ie Nd rae Nea WLS a Rogers, Hollis J., A new plant for the western hemisphere HOWL! Thm TASTE Sing (MOEN) "Solo ole udm Ue edna n ago coo ol velba Roofe, Paul, Cytogenesis within the supra-optic and paraventricular RNULEIMO PVE eg plo (aS track) ir sh ea Nate NUON atone onda Mia Schneiderhan, M. A., Spinal ganglion hypoplasia after limb aimputanon im the fetal, rat) (abStrach)) oc ast said Baca lay BCehelahy Ss MEPONey yAeich ie <4 Wess aches eH MMe ey ae nen atte os MORE Sergeant, Mary, Aquatic plants of Jefferson County, Kentucky. AMCMEMNVALOMSI (AIOSUGACE Pay i Je ee ARON GAD MS atl Skinner, J. T., Excretion of manganese by the rat .................. Influence of manganese intake upon urea excretion ...... Some functions of mineral elements in connection CG )aICE AN | LEXMA WTR ATE WC) 2 VON TKO) CD Mea ey SH oD. in Netti i Thirtieth Annual Meeting ....,... RITE AMUN OL PN it EN TROND Valieau, W. D., Colonies of Bacterium tabacwm on roots of SURE UG SEVER AM OMASSES( 14)00) So. tou Ah UN Ma NA RO a) Kield istrains Of tobacco mosaic (abstract), 2.0)... be Verhagen, Dirk, Preliminary studies on the separation of ether- TSOlubleNpronents! froin tobacco Wik .!.0) HAaMUMI eau nN Webster, Richard Curtis, The influence of nutrition on sporangial formation in Araiospora streptandra (abstract) ............... Wender, Simon H., Preliminary studies on the separation of cilerinsoluble )pioments, from: tobacco. ie eee. id al aie es epee ae CeCe y UNIV” tSITY OF KENTUCKY LIBRARY Lexington, Kentucky BONE 20 Se eee Library Form No, 7 — a eee ‘ea ae Dias ie Ley “NOH NNO 3 9088 01304 3187