ee oe [es QeOuhetceam euten Lf Ne. sahabwciete neodeenctterersncrs A % : tate ; .- : shademcsnatng oe | i 4 ; ane ok ly re : . hee Daa 3 ‘ See e es Spt ne Pn norm noy ee nn artes aT ae Me SAP Fe oe Fae UU ‘ ’ # bi; : ‘ a | i r ne bine 1G billy i mew hd > ar (TR a. 4 reac at an aA, ili Cee al ov dada, \) i Rabies “AN oily Aly as ee at , _ i ody : TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE Volume 21 — 1960 Published by THE KENTUCKY ACADEMY OF SCIENCE TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE EDITORIAL STAFF Editor Rocrer W. Barsour Associate Editors JoHn M. Carpenter, Zoology Barsara M. Conxin, Geology SETH GILKERSON, Bacteriology and Medical Technology Warp SuMPTER, Chemistry Mary E.. Wuarton, Botany Editorial Office Department of Zoology University of Kentucky Lexington, Kentucky CONTENTS OF VOLUME 21 No. 1-2 A Search for Long-Lived Ca®° and Cr56 WILLIAM D. EHMANN and JOHN R. HUIZENGA ...........cceseeeceeee Materials, Techniques and Methods in Teaching Psychology in 34 Secondary Schools Pee Nira mA CANIS oe mrt Mace deh ucts! Va ckttsiuascceuenadsuawatenenoiivaidaukedesseueee A Study of the Worm Snake, Carphophis amoenus Say, in Kentucky RRONGERY WV), SARBOU Ric csyastndvsavcersussuisnstandesacusoessensconssnduedtigartessszee The Chromosome Number of Helianthus decapetalus BD aaa es NAMM EN arc cere ce crc s ca caee dances sais cou'vdaceusazngusstatesemser seed Sugars in the Nectar of the Poinsettia, Euphorbia pulcherrima SISTER VirGINIA HEINEs and SisTER MARY ADELINE OLE LAT Ey SP al NSE ge As 9 ONAN a tg Ce Sulfur Compounds as Inhibitors in Oil Bearing Corrosion N. W. Hati, W. D. WitxiiaMs and J. R. MEADOW............0006 The Cave Snail, Carychium stygium Call PES Ee Ube EMU ERU CEU I xe cs anGvce=\ oases nsdsev soSeee eiscanenestscseesdsanecnetecesss¢sies Applications of Solar Energy Ma Vic HERA SAININIAG cc's cous enncot eves eds car coaduhcenasaCuuestnvetrenesscsscsaoesceetapazbae EMME A A EERBT SS aces cere ce eee pass even cea aah es nodes aneacaee vane cchnetenancscaanconsonnsonct No. 3-4 Soil Temperature Measurements at Lexington, Kentucky, from 1952 to 1956 E. B. PEnrop, J. M. ELuiotr, and W. K. BROWN ...............0008 Cyclic Entry-Exit for Subroutines ere SMES ccinicl slr Jie PIGNUAIN 5 ©, isc acecsaceesdtesscoucsce-anccucvearssavts The Salting Effect of Sodium Chloride on the Extraction of Erbium Acetylacetonate and 8-quinolinol Y. G. IsHma, J. F. STEINBACH, and W. F. WAGNER ..........00000 A North American Leafhopper Previously Confused with Typhlocyba andromache McAtee (Homoptera, Cicadellidae ) [PAL ag] FOr 027 oe Me ea Studies of Overwintering Potential of Certain Drosophila Species IN AITO CID. LPANU SON: sce saaceccestushtencaceosuiuaestuencacseaucnevaevvucsssceaso¥e Pape MINNA HEIN Siete cates cci4ia cu 4 < 10" years); Pue** (~ 8 10° years), and Cm?* (> 4 xX 10% years) appear to have half-lives in this range. Of these, only Pb?® which has a stable decay product, Tl°°°, would be important for isotopic dating. The other two radionuclides may have had, how- ever, important thermal effects in the history of the earth. A number of radionuclides that might be produced by two succes- sive neutron captures on the highest mass number stable isotope of a given element are still unknown. In this work a search was made for the two nuclides Ca°® and Cr°®. Previous work by Jones (1956) has established exclusion limits for the half-life of Cr°® of from 2 hours to 200 years. He believed it to be long-lived. Roy and Yaffe (1957) cat8 n cat? cones (Ema cal0 Stable Ms Bd 8.8 min ? Ca a a Se 57.2 min P 1.7 min Bie p49 450 Stable Stable Fig. 1. Production and decay of Ca50 * Based on work performed partly under the auspices of the U.S. Atomic Energy Commission. eT oct 10 08 2 William D. Ehmann and John R. Huizenga using a long pile irradiation of enriched chromium established that: tye Cr®®/c Cr®> = 270 years/barn if it is not very short. No published data is available on Ca®®. Both Ca®® and Cr®® were searched for in this work by use of the active daughter extraction technique. The production and parent daughter relationships for these nuclides are given in Figures | and 2. el Ma ee) ul ee Stable 3.52 min | 2 Mino? oe nee Stable: 2.58 hr RY FeD6 Stable Fig. 2. Production and decay of Cr®6 Experimental Procedures and Results Ca5®? Experimental—A 28.97 mg sample of CaCO; containing calcium enriched at Oak Ridge National Laboratory in Ca*® to 51.4% was irradiated in a high flux position at the Materials Testing Reactor at Idaho Falls, Idaho for approximately nine months. The total nvt received was 4.66 < 107! neutrons-cm—?. The irradiated sample was allowed to cool for one month before starting the chemistry. The radiochemical procedures consisted of solution of the sample in dilute HCl and extration of the Sc°® active daughter, using Sct? carrier, into a 0.5M TTA solution in benzene. The efficiency of the extraction as determined with Sc*® tracer was 97.5%. Since the half- life of Sc®® is only 1.7 minutes, rapid chemistry was required. The elapsed time from the start of the extraction to the start of the count- ing was always less than four minutes. The benzene layer containing the Sc-TTA complex was quickly separated and placed in a 7 ml volume plastic vial mounted in front of a 214” & 214” Nal crystal scintillation detector. The gamma spec- trum of the sample was examined for the 1.17 and 1.59 Mey gamma ray photo-peaks of Sc®° using a 256 channel pulse-height analyzer. A Search for Long-Lived Ca*® and Cr*® 3 The physical geometry of the counting set-up was approximately 20%. The initial gamma spectrum obtained for a sample was compared to a “background” spectrum taken 15 minutes after the initial count in order to detect any activity in the region of the two Sc°® gamma ray photo-peaks. Results—A series of five Sc extractions were made on the irradiated calcium sample in an attempt to detect the Sc°*® active daughter of Ca®°. In no case were peaks observed at 1.17 or 1.59 Mev and no difference outside of the statistical standard deviation was observed between the first count and the final count on a given extraction. The background counting rate in the vicinity of the 1.17 Mev photo-peak was ~ 8 counts/minute/channel and in the vicinity of the 1.59 Mev photo-peak was ~ 1 count/minute/channel. Assuming a counting rate for Sc®® less than or equal to the stand- ard deviation of the initial counting rate in the area bracketing the 1.59 Mev peak, the following ratio is obtained: tio Gall /o Ca? == £ SC 10? years/ barn If the cross section of Ca*® is estimated to be approximately 0.1b, a lower limit to the half-life of Ca°° of approximately 1 x 10° years is obtained. The possibility that Ca®® might be very short lived is, of course, not excluded. Cre Experimental—A 36.695 mg sample of Cr.O3 containing chromium enriched at Oak Ridge National Laboratory in Cr°* to 83.1% was irradiated in a high flux position at the MTR reactor for approxi- mately nine months. The total nvt was also 4.66 10?! neutrons-cm—?. The sample was allowed to cool for 2 months before the start of the chemistry. The radiochemical procedures consisted of fusion of the sample in Naz,Ovz followed by solution in dilute HCl. The acid solution was treated with a few drops of HzO. and evaporated to near dryness. The residue was then made up to 15 ml with H:O and scavenged with CuS. Cr(OH)s; was separated by precipitation with NH,OH and dis- solved in 15 ml of concentrated HNO3. Mn? carrier was added and solid KCI]O3 added slowly with heating to affect oxidation of Mnt? to MnO, and Crt? to CroO;—?. The MnO. was filtered off and dis- carded. This MnO, precipitation step was repeated several times using Ir carrier to remove Ir!” contamination, which follows this Mn chemistry. The Cr,0;—* was reduced to Crt+® between MnOz pre- cipitations by addition of concentrated HCl and a few drops of H2O, ST 4 William D. Ehmann and John R. Huizenga followed by heating. Zr®® contamination was removed by scavenging with BaZrF.. The purified Cr sample at this point showed only Cr°! activity in a gross gamma ray pulse-height analysis. This sample was then used to attempt the extraction of the Mn*® active daughter of Cr°®. The method used was addition of a known amount of Mn? carrier to the purified Cr+* solution followed by oxidation to MnOs, as before. This MnO, which carried any Mn°® present was reprecipitated several times as above with inactive chromium carrier to remove traces of Cr®!. Solution of the MnO, between precipitations was affected by use of concentrated HNO3 and a few drops of H2,Oz. The final MnO, precipitate was filtered on a weighed paper disk, weighed to deter- mine chemical yield, and mounted on a card for beta counting of the 2.58 hour half-life Mn°°. The time required for the chemistry was about two hours. Counting was done on the first shelf of a standard end-window proportional counter having a background of 10.8 c/m. A 85.5 mg/cm? Al absorber was used to eliminate a high background of unidentified low energy betas. Results—A series of four MnO, samples were prepared as above. The samples were counted at short intervals for approximately six hours. In no case was any decrease in the counting rate observed corresponding to the 2.58 hour decay of Mn°*. In the best run the background corrected counting rate of the MnO» sample through the absorber was 35 c/m. No variation in this counting rate outside of the statistical standard deviation was observed over a period of more than five hours. If the counting rate of Mn*® at the time of the initial count is taken to be less than or equal to the standard deviation of the counting rate for the initial ten minute count, the following ratio is obtained: tijo Cr®®/o Cr®> = 1100 years/barn If the « Cr® is estimated to be approximately 10 barns, then the lower limit for ty/2 Cr°® is approximately 1.1 x 10+ years, if it is not very short lived. Since Jones (1956) has found that the half-life of Cr** is not in the range of 2 hours to 200 years, the exclusion limits for the half-life of Cr°® may now be extended to from 2 hours to approxi- mately 1.1 x 10* years. The possibility that Ca®®° and Cr°® might have half-lives in the range specified for extinct natural radionuclides is, therefore, not ex- cluded by this work. In order to finally extend the exclusion limits through this half-life range, much longer irradiation times and larger, highly enriched samples will have to be used. A Search for Long-Lived Ca** and Cr’® 5 Literature Cited Kohman, T. P., 1956. Extinct natural radioactivity: Possibilities and potentialities. Ann. N. Y. Acad. Sci. 62:503. Jones, J. W., 1956. Spallation yields from chlorine with 45-430 Mev. protons; A search for unknown medium-light even-even nuclides; and beta lifetime sta- tistics. U.S. Atomic Energy Commission Document NYO-6627. Roy, L. P. and Yaffe, L., 1957. Search for successive neutron capture reactions on Mg?6, Si80, and Cr54. Can. J. Chem. 35:176. Accepted for publication December 4, 1959. MATERIALS, TECHNIQUES AND METHODS IN TEACHING PSYCHOLOGY IN 34 SECONDARY SCHOOLS PAUL McNEELY Asbury College, Wilmore, Kentucky Introduction After a brief introduction, this article will present the reactions of students and teachers to practices in current use in psychology classes in Indiana secondary schools. The procedure and major findings of the author’s doctoral thesis relative to materials, techniques and meth- ods will be given with some implications that they may have for Ken- tucky high schools. According to T. L. Engle! Kentucky and Arkansas are the only two states in which 25 percent of the secondary schools offer a course in psychology. A letter from the Department of Education of Ken- tucky? states that of the 420 public high schools, 101 offer a course labeled Social Psychology. Of the 68 non-public schools, 6 offer a course with the above title. Dr. Halice Wiggs? found in his doctoral study that 82.7 percent of the principals of Indiana are in favor of the course. Actually 14 per- cent of the schools of this state offer the course. Nationally, from the article by Engle referred to above it was learned that over 1,082 high schools offer the course. This is 8.4 percent of the total of 12,939 high schools reported by officials in 34 states. The American Psychological Association has a special committee working on the problem at the secondary level. Since one fourth of the high schools of Kentucky offer a psychology course it would seem of interest and value to present the major findings of the Indiana study conducted in 34 high schools. It is hoped that some of the findings will be helpful to those in Kentucky who are concerned with teaching or curriculum construction. The writer's study* was conducted during the spring of 1956. A stratified sampling of 34 Indiana high schools in a 30 mile strip north and south and east and west was made. Personal visits were made to schools and tests given to 1,236 students (715 females and 521 males) who were enrolled in psychology courses. The 88 teachers in the 34 schools visited were mailed questionnaires before visitation. These were collected at visitation and later analyzed. One teacher did not return his questionnaire making a 97.4 percent return. Materials, Techniques and Methods in Teaching Psychology 7 Major Findings of the Thesis By confining our discussion to the data presented in this thesis, it is possible to summarize as follows: Major findings related to problems of students. Most students of high school psychology thought the course had been helpful, but they did not feel free to discuss their personal problems with their teacher. One reason for this could have been that most psychology teachers had only five hours or less of free time per week for personal counsel- ing. Teachers tended to overestimate the degree of help students re- ceived on their problems. Students checked Reader's Digest, Life, and Colliers as the magazines most valuable in solving their personal prob- lems. The outstanding problem on which the students desired help but did not obtain it was choosing a career. Not only were more females enrolled in the psychology course, but they also apparently received more help from the course. Major findings related to topics. The topic liked least by students was how to study. The largest percentage of students indicated that the topics misconceptions about psychology, and how to study, should be used less than at present. On the other hand, students desired more use of such topics as courtship and marriage, prevention of mental disease, and emotions and their control. The favorite topic of students was personality. A higher percentage of students indicated that this topic had helped them more than any other topic. Topics of highest interest were distributed throughout the course. Major findings related to materials. The text most frequently used was by Engle, Psychology: Its Principles and Applications, second edi- tion, in a course which was almost always labeled Psychology. Teach- ers rarely used a syllabus, but almost half of them used supplemental reading in addition to the text. The least frequently used materials were recorders, slides, World Book, television, health charts, and mimeographed outlines while the most frequently used materials were filmstrips, reference books, study questions, newspapers, and maga- zines. Students desired more use of films and filmstrips, and less use of World Book, health charts, and mimeographed outlines. Major findings related to methods. The most unpopular method ac- cording to students was the writing of an autobiography. Dramatiza- tion and skits were not very popular. Both teachers and students agreed that by far the most frequently and best used methods were lecture and discussion. When outside speakers were used, they were more frequently a psychologist or a social worker. Some teachers did not use any speakers but one used as many as 10 different persons. 8 Paul McNeely Both teachers and students agreed that more outside speakers should be used in the course. Experiments performed as demonstrations and those performed by individual students were power of observation, reaction time, learning time, mental telepathy, and conditioned response. The tests most frequently explained to students by teachers were Rorschach, Kuder Preference Record, Draw a Picture Test, and Cali- fornia Test of Mental Maturity. Tests most frequently administered to students were Kuder Preference Record and California Test of Mental Maturity. Major findings related to the evaluation of the course. Most teach- ers gave a combination of objective and essay tests. An opportunity was given for discussion of examinations. Both teachers and students preferred announced written quizzes and chapter tests. Teachers gave these types of examinations. Unannounced quizzes were not popular with either teachers or students. Teachers thought the psychology course was average in difficulty; students tended to think it was easy. Major findings related to the high school psychology teacher. Psy- chology teachers tended to be enthusiastic about their work. They desired summer workshops, magazine articles, seminars, and other aids that would help them in presenting psychology at the secondary level. Those teachers who used a variety of teaching methods seemed to have better rapport with their students as judged by their students’ willingness to discuss personal problems with their teacher. The size of the school, class, number of years psychology had been taught did not seem to be as important as the relationship that existed between the student and the teacher. Even the type of materials and methods used seemed to be secondary to the personality of the teacher. Very few psychology teachers were rated below average in their ability to present materials and methods in high school psychology. implications If it can be inferred that Indiana and Kentucky present analagous situations as far as psychology at the secondary level is concerned, then it would seem that more attention should be given to the prepa- ration of high school psychology teachers. This seems to indicate that more free time should be given for personal counseling, more use of outside speakers, and an increased volume of psychological informa- tion that is scientific yet adapted to the use of high school psychology teachers. Materials, Techniques and Methods in Teaching Psychology 9 Summary Results from a survey of the teaching of psychology in 34 high schools are presented, together with some of the possible practical applications of the findings for Kentucky high schools offering a course in psychology. Literature Cited l. Engle, T. L., 1952. “Teaching of Psychology in High Schools,” Amer. Psych., Tol. Department of Ed., Frankfort, Ky., dated Oct. 19, 1959. 3. Wiggs, H., The Status of Psychology in Indiana Secondary Schools. Indiana University, Bloomington, 1952. 4. McNeely, P. Materials, Techniques, and Methods in Teaching Psychology. In- diana University, Bloomington, 1958. bo Accepted for publication November 27, 1959. A STUDY OF THE WORM SNAKE, CARPHOPHIS AMOENUS SAY, IN KENTUCKY ROGER W. BARBOUR Department of Zoology, University of Kentucky, Lexington, Ky. The worm snake, Carphophis amoenus Say, ranges from southern New England south to central Georgia and Alabama and west to east- ern Nebraska, Kansas, Oklahoma, and Texas. In the Mississippi val- ley, it extends southward to the Gulf. Three subspecies are currently recognized, C. a. amoenus, C. a. helenae, and C. a. vermis. The latter is essentially confined to the area west of the Mississippi river, while the preceding two are found east of the Mississippi. Carphophis a. helenae ranges from the Mississippi eastward to the Appalachians; C. a. amoenus ranges in the Appalachians and eastward to the coast. (Conant, 1958). The three subspecies may be separated by the fol- lowing key: A. Pink of belly extending up sides onto third scale row; color aboverusually.punplisiinlacka senecue eee C. a. vermis (Western Worm Snake) AA. Pink of belly extending up sides onto only first or second scale row; color above usually plain brown. B. Internasals and prefrontals distinct .............. C. a. amoenus (Eastern Worm Snake ) BB. Each prefrontal fused to the corresponding internasal ........ C. a. helenae (Midwest Worm Snake) The species ranges throughout the state of Kentucky. Schmidt (1953) stated that C. a. amoenus occurs “. . . east of the Appala- chians,” which implies that only one subspecies, Carphophis a. helenae Kennicott, occurs in Kentucky. The 116 specimens of Carphophis amoenus from Kentucky in the zoological collections of the University of Kentucky have been ex- amined. Measurements, scale counts, condition of head plates, overall dorsal coloration, sex, number and size of eggs, if present, date and locality collected, and habitat, where known, were recorded for each specimen. These data are summarized in this paper. The specimens were largely collected by the author during the past eight years. The Research Fund Committee of the University of Kentucky provided financial assistance incident to the collection of many of the specimens. ! A Study of the Worm Snake 11 Distribution in Kentucky Previous literature records, substantiated by specimens I have ex- amined, leave no doubt that both C. a. amoenus and C. a. helenae, as currently understood, occur in Kentucky. Fig. 1. Fig. 1. The distribution of Carphophis amoenus in Kentucky. Hexagons represent records of C. a. helenae; circles, C. a. amoenus; triangles, intergrades. Solid symbols represent specimens examined; hollow symbols, litrature records. Carphophis amoenus amoenus Say. Literature records are: “throughout Kentucky”—( Garman, 1894); Breathitt county (Dury and Williams, 1933); Bell, Harlan, and Letcher counties (Barbour, 1950b); Harlan and Letcher counties (Barbour, 1950a); Breathitt county (Bush, 1959). I examined 44 worm snakes that are referred to C. A. amoenus. They were collected from the following counties: Bell, 1; Breathitt, 21; Harlan, 21; and Lee, 1. Thirty-four of the specimens have the internasals and prefrontals separate, 6 have them fused, and 4 are intermediate in this characteristic. Carphophis amoenus helenae Kennicott. Literature records are: “throughout Kentucky”"—(Garman, op. cit.); Anderson and Fayette counties—( Funkhouser, 1925); Edmonson, Hardin, and Jefferson coun- ties—( Burt, 1933); Edmonson county—( Bailey, 1933); Carter county (Dury and Williams, op. cit.); Edmonson county (Hibbard, 1936); Fulton county (Parker, 1939); Rowan county (Welter and Carr, 1939); Oldham, Larue, and Warren counties (Barbour, 1950b). Thirty-four of the specimens examined are referable to C. a. helenae. They were taken from the following counties: Anderson, 1; Clark, 1; Edmonson, 6; Garrard, 4; Green, 2; Jessamine, 9; Menifee, 2; Nelson, 3; Russell, 1; Todd, 4; and Warren, 1. Twenty-six of the speci- mens have the internasals and prefrontals fused; five have them separ- ate, and four are intermediate. IB, Roger W. Barbour Although here considered C. a. helenae, the four specimens from Todd County (the westernmost specimens examined) are unlike any other Carphophis examined in the degree of contrast between the back and belly color, strongly resembling C. a. vermis. They differ from vermis, however, in the fact that the belly color extends only onto the second scale row. The specimens from Edmonson County show this marked contrast between back and belly color to a lesser degree. Additional collecting in western Kentucky will possibly reveal the Carphophis population there to be intermediate between C. a. helenae and C. a. vermis. Fig. 2. Photographs of a series of heads of Carphophis amoenus from Kentucky. A typical C. a. amoenus; B-H, intergrades between C. a. amoenus and C. a. helenae; |, typical C. a. helenae; J, individual with partly fused internasals; K, individual with partly fused — prefrontals; L, individual with internasals and prefrontals fused into a single large plate. A Study of the Worm Snake 13 B C J Fig. 3. Tracings from the photographs shown in Fig. 2. Carphophis amoenus amoenus Say X Carphophis amoenus helenae Kennicott. Barbour (1950b) has pointed out that “along the western edge of the mountains a tendency toward intergradation appears.” Dury and Williams (1933) mentioned a specimen, under the name helenae, from near Barbourville, Knox county, that has “the internasals and prefrontals fused on the left and separate on the right.” Thirty-eight specimens have been examined that are considered intergrades. They were collected from the following counties: Bath, 6; Carter, 9; Fleming, 10; McCreary, 9; Rowan, 1; and Whitley, 3. Fourteen of these have the internasals and prefrontals separate; 19 have them fused; and 5 are intermediate. Apparently a change is occurring in the Carphophis population of Rowan and adjacent counties. Welter and Carr (1939) stated “among 14 Roger W. Barbour all Carphophis examined in eastern Kentucky no typical amoena were found.” Barbour (1950b) pointed out the presence of typical amoena and intergrades in the area. In this study, 26 specimens from the ad- joining counties of Carter, Rowan, Bath, and Fleming, collected since the appearance of the paper of Welter and Carr, were examined. Six- teen of them exhibit fused internasals and prefrontals; in 7 they are separate, and 3 are intermediate. These data indicate that these coun- ties are in the range of intergradation between the two subspecies. Intergrades between the two subspecies are quite variable, repre- senting practically all possible stages in the degree of fusion of the internasals and prefrontals. Occasional specimens are found with fused internasals or prefrontals. In one case, the internasals and prefrontals were fused into one large plate. Some of these degrees of fusion are shown in Figs. 2 and 3. Habitat Worm snakes in Kentucky are almost invariably found beneath some object, generally a rock or a log. They may be found in a great variety of habitats, ranging from virgin timber through brushy areas to weed-grown fields, and even open pastures. They are, however, rarely found far from a wooded area. Food Of the 116 specimens examined, only 22 contained food. In every case, only earthworms, along with the detritus from the alimentary canal of the worms were found. Barbour (1950a) reported only earth- worms from the stomachs of 10 of 22 individuals collected in Harlan county, Kentucky. Reproduction A total of 20 egg-bearing females was examined, all taken between February 4 and June 18. Average egg number per female was 2.6, varying from 2 to 5. Average egg length in February (4 eggs) and March (2 eggs), was 5mm.; in April (19 eggs), 7.3mm.; in May (18 eggs), 15mm.; and in June (10 eggs), 18.6mm. Blanchard (1925) re- ported that 5 captive individuals laid eggs between July 4 and 11. Bush (1959) reported that in Breathitt county, Kentucky, “Egg laying seemed to take place about mid-June. Seven females having deposi- tional-sized eggs had an average clutch of 3.2; the average egg length was 24.2mm. Ten females taken between June 19 and July 2 had no eggs.” Funkhouser (1945) stated that Carphophis amoenus “lays eggs in late summer.” Data herein presented indicates that in Kentucky, egg-laying occurs about the middle of June. All females of egg-laying size (herein indicated to be a minimum of about 185mm. in total A Study of the Worm Snake 15 length) taken prior to June 10 contained enlarged eggs. No adult female taken after June 18 contained enlarged eggs. Thirty-three adult females averaged 233.4mm. in total length, ranging from 185 to 282mm. The average of those with eggs (20 specimens) was 233.4; those without eggs (13 specimens) averaged the same. Table 1. Table 1.—Number and Size of Eggs in Carphophis amoenus from Kentucky (Measurements are in mm.) Number Females Length Length of Length of eggs fe) with of egg- non-egg- Number (mean and Date females eggs bearers bearers of eggs extremes ) Feb. 4 2 2 948,948... 2.2 5 (5-5) March 14 1 1 PREVA GT Ser 2, 5 April 8 2 2 1651090 i. 2.3 8.5 (7-10) April is * 2 2 DAS ORIEN Naess 2.5 2.5 (2-8) April 23 1 1 PAPA GH” tN rian een 2, 13 April 24 1 1 DAC Wee 2 13 Apml 28 2 2) BINGL OMS Ys eapeers TE, 9 (8-10) May 3 1 1 DAS fi a wiassevres 3 12 May 9 3 3 228,236,255 ........ 2.92.3 16.6 (14-19) May 10 2, 2 ZAIBIAG er: 3,4 15.5 (14-17) June 10 1 1 DAO all eg assSece 3 13 June 18 1 OW” 2 Ze DED TO TE Nee NS Recess June 18 3 2 237,260 270 3,4 21 (20-22) June 20 1 Ong a katas DOO Ewa VTA O DEEeRT seeks June 25 1 Ole ume ests. 283i) — sue De ake. ete June 28 4 ees ssacese QOGQUAIB8OQOG) seecesti to emicenceneeence July 2 2 OPE fsa DOO206) ye GF eeesd .. tacos Aug. 5 Ht Of a sce: WET ip PE). “hw cbcecdyuly pili desevsaeeees Aug. 18 1 Crmy es DSA oy Gu een cee ae Aug. 24 1 On es 3 ese3 DAT. i | aasescin -nssteseeeets Sexual Dimorphism Measurement of the total length and tail length of 48 females and 65 males revealed that the males have a proportionally longer tail than the females. Relative tail length (tail length ~ total length) was calculated for the specimens. There was no appreciable change with increase in total length. In the females, the tail averaged 14.4 percent of the total length, with extremes of 12.9 to 16.1 percent. In males, the percentage ranged from 16.4 to 20.5, averaging 19.1. This differ- ence is sufficiently great to be readily discernible to the experienced eye. Summary One hundred sixteen specimens of Carphophis amoenus from Ken- tucky were examined. Food consisted entirely of earthworms. Egg- laying occurred in the middle of June, and the average number of eggs was 2.6, varying from 2 to 5. Measurements of the tail and total 16 Roger W. Barbour length of 48 females and 65 males revealed that the tail of the females averaged 14.4 percent of the total length, whereas that of the male averaged 19.1 percent. The worm snake of the southeastern moun- tains of Kentucky is C. a. amoenus; that of central and western Ken- tucky is C. a. helenae. Intergradation between the two subspecies occurs along the western border of the mountains of eastern Kentucky. There is some evidence that worm snake of western Kentucky is inter- mediate between C. a. helenae and C. a. vermis. Literature Cited Bailey, Vernon. 1933. Cave Life of Kentucky. Am. Midl. Nat. 14(5): 383-635. Barbour, Roger W. 1950a. The reptiles of Big Black Mountain, Harlan County, Kentucky. Copeia (2):100-107. & 3 1950b. The distribution of Carphophis amoena in Kentucky. Copeia 3) :287. Blanchard, Frank N. 1925. A collection of amphibians and reptiles from southern pigiana ced adjacent Kentucky. Pap. Mich. Acad. Sci., Arts, and Letters. Burt, Charles E. 1933. A contribution to the herpetology of Kentucky. Am. Midl. Nat. 14(6) :669-679. Bush, Francis M. 1959. The herpetofauna of Clemons Fork, Breathitt County, Ken- tucky. Trans. Ky. Acad. Sci. 20:11-18. Conant, Roger. 1958. A Field Guide to Amphibians and Reptiles. Houghton Miffin Co., Boston XV + 366 pp. Dury, Ralph and Raymond S. Williams. 1933. Notes on some Kentucky amphibians and reptiles. Bul. Baker-Hunt Foundation Mus. 1:1-22. Funkhouser, William D. 1925. Wildlife in Kentucky. Ky. Geol. Surv., Frankfort, Kentucky 1-385. . 1945. Kentucky Snakes. Dept. Uni. Ext., Uni. Ky., Lexington. 1-31. Garman, Harrison. 1894. A preliminary list of the vertebrate animals of Kentucky. Bul. Essex Inst. 26:1-63. Hibbard, Claude W. 1936. The amphibians and reptiles of Mammoth Cave Na- tional Park Proposed. Trans. Kans. Acad. Sci. 39:277-281. Parker, Malcolm V. 1939. The amphibians and reptiles of Reelfoot Lake and vicinity, with a key for the separation of species and subspecies. Jour. Tenn. Acad. Sci. 14(1):72-101. Schmidt, Karl P. 1953. A checklist of North American amphibians and reptiles. Am. Soc. Ichth. and Herps. VIII + 280 pp. Welter, Wilfred A. and Katherine Carr. 1939. Amphibians and reptiles of north- eastern Kentucky. Copeia (3):128-130. Accepted for publication December 15, 1959. THE CHROMOSOME NUMBER OF HELIANTHUS DECAPETALUS? DALE M. SMITH Department of Botany, University of Kentucky, Lexington In an earlier paper (Heiser & Smith, 1955) on chromosome num- bers in Helianthus, it was reported that H. decapetalus L., was an apparent tetraploid with a chromosome number of N=34. This report was based upon material collected in the vicinity of Bloomington, Monroe Co., Indiana. Subsequently, new cultures of H. decapetalus were obtained from other localities which proved to be diploid CN==17 ). Methods and Materials Plants from which chromosome counts were obtained grew in ex- perimental areas at Indiana University and the University of Ken- tucky, as well as natural habitats. Counts were obtained from micro- spore mother cells in each case, and in a few plants additional deter- minations were made using root-tips. Buds for meiotic studies were obtained at the proper age and fixed in 95% ethyl alcohol and glacial acetic acid (3:1) for approximately 24 hours and then transferred to 70% ethyl alcohol for storage. Most observations were made using temporary aceto-carmine squashes, but a few slides were made perma- nent for future reference and record. Root-tip material was obtained from potted plants in the green house. A pre-treatment was found to be necessary to study adequately the mitotic chromosomes. Best re- sults were obtained using 0.1% aqueous colchicine for four hours at approximately 4°C., or a mixture of 0.1% colchine and 0.02M 8-quino- linol (1:1) also for four hours at 4°C. The former treatment was some- what better for mere counting, but the latter was superior for studies of chromosome morphology. Staining was by the Feulgen technique and aceto-carmine. Voucher herbarium specimens are on deposit at Indiana University and the University of Kentucky. The chromosome counts obtained are listed in Table 1. Conclusions The widespread occurrence of both diploid and tetraploid popula- tions of H. decapetalus is clearly indicated by the results presented in Table 1. Comparable situations have been reported in a number of other plant genera, but this is the first instance of this type in the genus Helianthus. It is rather unusual that similar situations have not 1 Supported in part by NSF grant G-6380. 18 Dale M. Smith Table 1.— Chromosome Numbers of Helianthus decapetalus L. N-chromosome State and County number Collector and number Maine Somerset County Ly, Smith C-5 Vermont Bennington County 172 Eaton 565 New York Chautauqua County 84 Heiser P-190 Pennsylvania Crawford County 84 Heiser 4558 Virginia Rockingham County 17 Smith P-412 Smyth County 17 Martin s.n. West Virginia Randolph County 17 Smith P-406 Indiana Brown County 34 Heiser P-299 Lawrence County 84 Smith 1050 Monroe County 842 Heiser 3005 Monroe County 342 Heiser 3017 Monroe County 84 Smith P-272 Monroe County 84 Smith 1071 Kentucky Breathitt County 34 Smith K-115 Martin County 17 Smith C-1 Wolfe County 84 Smith K-122 been discovered in other species of Helianthus in light of the high degree of polyploidy which has been previously reported (Heiser & Smith, 1955). The closest approach is seen in the complex presently treated as H. strumosus L., in which both tetraploids (N=84) and hexaploids (N=51) are known. As yet, I have not been able to separate the two cytodemes of H. decapetalus morphologically. If differences exist, they seem to be largely quantitative, with the tetraploids being somewhat more robust than the diploids. Studies are also in progress dealing with the prob- lem of autopolyploidy vs. allopolyploidy in this and related species of Helianthus. The available evidence strongly suggests that tetraploid H. decapetalus may be a true autopolyploid, but the final determina- tion must be based upon more data than is presently available. Summary A perennial sunflower, Helianthus decapetalus L., originally re- ported to be a tetraploid (N=384), is shown to include diploids 2JT am indebted to Dr. C. B. Heiser, Jr., of Indiana University, for these counts. The Chromosome Number of Helianthus decapetalus 19 (N17) as well. Tetraploids are reported from ten localities, while diploids are now known from six stations. The two cytological types are closely similar morphologically, and it seems possible that this may be a case of true autopolyploidy. Literature Cited Heiser, C. B. Jr., and D. M. Smith. 1955. Proc. Indiana Acad. Sci. 64:250-253. Accepted for publication December 15, 1959. SUGARS IN THE NECTAR OF THE POINSETTIA, EUPHORBIA PULCHERRIMA* SISTER VIRGINIA HEINES and SISTER MARY ADELINE O’LEARY Nazareth College, Louisville, Kentucky Very few determinations on the individual sugars in flower nectars have been carried out. The older analytical methods required large volumes, which were not available in the nectars investigated. By using the technique of paper chromatography, it is now possible to determine the sugars present in micro-amounts of material. Previous work done on nectars by Wykes (1) has shown that sucrose, glucose and fructose are not equally attractive to bees. He analyzed (2) the nectars from twelve species of flowers by paper chromatography and found that the proportions of glucose and fructose varied greatly in different species. The present study investigates the total sugar con- tent of the fresh nectar of the Poinsettia, and the separation of the individual sugars by paper chromatography. Experimentation Total Sugars by the Anthrone Method (3) Four hundredths of a milliliter of the clear, viscous nectar were drawn from the yellow gland of the cyanthium and diluted with dis- tilled water to 2 ml. Three milliliters of the anthrone reagent were pipetted into each of three test tubes, followed by 0.2 ml. of a stand- ard glucose solution (0.02 mg./ml.) into the first tube, 0.2 ml. of dis- tilled water into the second tube as a blank, and 0.2 ml. of the diluted nectar into the third tube. The solutions were mixed by tapping and tubes placed in a thermostatically controlled water bath at 80°C for 10 minutes, then cooled under tap. By using Beckman B and Cole- man (6C) spectrophotometers the optical density was read at 640 mu. Individual Sugars 1. Circular Chromatograms. The circles of Whatman No. 1 paper (32 cm.) were spotted with known sugars and nectar solutions, placed in desiccators with strings for wicks, and let run for 18 hours in n-butanol-pyridine-water, 6:4:3, solvent. They were removed, air- dried, and dipped into an aniline phthalate reagent. After drying for several hours at room temperature the chromatograms were heated for 5 minutes at 105°C and then 10 minutes longer at 120°C to bring out the sucrose (4). Examination under ultraviolet light helped to identify the sugars. * Supported in part by grant from National Institutes of Health. Sugars in the Nectar of the Poinsettia 21 2. Sheet Chromatograms. Sheets of Whatman No. | paper, 23 « 23 cm., were spotted with known sugars and nectar samples using 0.01 ml. volumes. After 24 hours in the solvent, the silver nitrate method was used to determine the sugars (5). Quantitative analysis of the nectar was made by the Pridham method (6). Standard solutions of pure glucose and fructose were spotted in 0.01 ml. volumes so as to extend over a range of 50y to 175y by weight. The chromatograms were developed with the 6:4:3 solvent for 24 hours, air-dried, dipped into the p-anisidine hydrochloride reagent, and heated at 180°C. for 10 minutes. The colored spots and blanks were cut from the paper and eluted by shaking 5 minutes with 3 ml. of the methanolstannic reagent. Optical density values read at 640m» were plotted against the weight of standard glucose and fructose solutions. The standard curves were used to determine the amount of glucose and fructose in the nectar sample. Several drops of the viscous nectar were expelled from the yellow gland onto a watch glass and kept in an airtight box in a warm room for a period of six months. The sugars crylstallized out and became embedded in a clear solid. Chromatograms were run on these crystals and the ratio of glucose to fructose calculated. Results and Discussion The total sugar content of the nectar determined by the anthrone method was found to be 50% calculated as glucose: 0.26 O.D. (nectar) 0.56 O.D. (Std. glucose) The original sample (0.04 ml.) was diluted to 2 ml.; 0.5 ml. of this was made up to 10 ml. and 0.2 ml. of this last dilution was used to determine the total sugar content. Circular chromatograms made on samples of the fresh nectar showed presence of glucose, fructose and sucrose. The crystalline product indicated glucose and fructose only. Zimmerman (7) found small amounts of the enzyme, invertase, in the nectar of the Poinsettia, Euphorbia pulcherrima, which could account for the absence of su- crose in the crystalline sample. Calculated values of these from the standard curves gave 46y of glucose and 35y of fructose, out of a spotting of 82y sample. The crystals were of the orthorhombie type (8). These results are summarized in Table 1. The ratio of glucose to fructose (G/F) was found to be 1.39, which corresponds closely to that of one species of another plant investigated by Wykes. | 10 < 0.02 mg.(Std.) x —— —=5 mg./10 ml. 0.2 22, Sister Virginia Heines and Sister Mary Adeline O’Leary Table 1.— Sugars in Crystallized Nectar of the Poinsettia Sugars Volume applied to Optical Weight sugar found Chromatogram Density OF Glucose (1) 10 zl 0.210 47 (2) 10 zl 0.200 45 Average 46 Fructose (1) 10 zl 0.090 34 (2) 10 wl 0.095 36 Average 85 Total sugars in crystalline nectar 81 Total sugar calculated from sample weight 82 Literature Cited Wykes, G. R., J. Exp. Biol. 29, 511, 1952. Wykes, G. R., Biochem. J. 53, 294, 1953. Roe, J. H., J. Biol. Chem., 212, 355, 1955. Block, R. J. et al., “Paper Chromatography and Paper Electrophoresis”, 2nd ed., Academic Press, N. Y. 1958. 5. Green, S. R., and Stone, I., Wallerstein Laboratories Communications, 15, 850, 1952. 6. Pridham, J. B., Anal. Chem., 28, 1967, 1956. 7. Zimmermann, M., Ber. schweiz. botan. Ges. 63, 402, 1953. Chem. abst., 48, 11575», 1954. 8. Wycoff, R. W. G., “Structure of Crystals”, 2nd ed., Chemical Catalog Co., N. Y. 1931. Cfo i) Accepted for publication October 27, 1959. SULFUR COMPOUNDS AS INHIBITORS IN OIL BEARING CORROSION N. W. HALL,1 W. D. WILLIAMS,? and J. R. MEADOW Department of Chemistry, University of Kentucky, Lexington, Ky. Reactions in an internal combustion engine are varied, complex and uncertain. Lubricating oil temperatures may range from 120°C to 250°C. Oil is whipped around as a mist in the crankcase and ab- sorbs considerable quantities of air. Under these conditions, lubri- cating oil is subject to increased oxidation and breakdown. The break- down processes are necessarily very complex. The order of events and the intermediate reaction products depend on such factors as structure of oil molecules, temperature, pressure, presence of air, inhibitors, metallic catalysts, and others. Such a combination of fac- tors introduces a considerable element of uncertainty into any specu- lation about the nature of breakdown products (1). The formation of lacquers and sludges as well as the corrosion of bearings is usually attributed in part at least to the attack of the oil by oxygen (2). Metals are not usually corroded by the hydrocarbon components per se which make up a very large portion of any lubri- cating oil, but corrosion does occur under certain conditions as the result of the presence of impurities or “catalysts” in lubricants and as a result of the development of oil oxidation products. Lubricants can vary as much as a hundredfold in their bearing corrosion tendencies. Bearings themselves differ greatly as to their resistance to corrosion in oils. Copper-lead and cadium-base bearings have greater me- chanical strength than babbit bearings, but unfortunately they are more subject to chemical attack at elevated temperatures by oxidation products of the oil. Thus much work has been devoted to the problem of finding so-called additives which will suppress the oxidation of the oil and thereby decrease the harmful effects on the engine and bear- ings as well as increase the life of the oil. The literature on the breakdown of lubricants reveals many differ- ent viewpoints regarding the mechanism of oil oxidation and the role of antioxidants or inhibitors present in an oil (3). Numerous investi- gators have studied the effects of various types of compounds on the stability of oils when they are subjected to oxidative conditions (2-15). In most instances the extent of oil oxidation, with or without inhibi- tors, has been reported in terms of oxygen absorbed, acidity developed, the amount of sludge formed in the oil after a given time, and to some extent the corrosion of a metal surface. 1 Present address, Shell Oil Company, Houston, Texas. 2 Present address, Harding College, Searcy, Arkansas. 24 N. W. Hall, W. D. Williams, and J. R. Meadow The only completely satisfactory test on a lubricant is necessarily long, tedious and expensive. Such a test would be a long engine test, and this would be followed by a thorough road test and examination of engine parts thereafter. Obviously, many shorter screening tests have been devised to help arrive at some conclusions about the stabil- ity of oils and what inhibitors should be used in them. It is known that lubricating oils contain many naturally occurring sulfur compounds which may function as oxidation inhibitors (6). Depending on the source of the oil, the amount of sulfur compounds present may vary from a trace to as much as twenty percent (6). Since lubricating oils are so complex, it is impossible to determine the exact structure of the sulfur compounds present. As suggested by Denison and Condit (7) it is perhaps better to study the problem of sulfur inhibition of oils by the use of known sulfur additives and not- ing their effect on the oxidation of a base stock oil of known sulfur content. These authors studied the inhibiting effects of several kinds of sulfur compounds on the oxidation of completely desulfurized oils. They concluded that thioethers, having at least on aliphatic or cycloali- phatic group, were the most effective inhibitors tried. Their findings were based on oxidation rates using a Dornte type of apparatus (8) for measuring the volume of oxygen absorbed by a given sample of oil. The purpose of our investigation was to study organic sulfur com- pounds as corrosion inhibitors, or possibly corrosion catalysts, and note the actual effects produced on a bearing strip. The choice of bearing strip was that of a cadmium silver metal surface.? The test used was similar to the one described by Moran, Evers and Fuller (17) which is an air blowing test for indicating the corrosive effect of oils on cadmium silver bearing surfaces. This type of short test is used in some laboratories as a preliminary screening test for studying oil addi- tives. Because of the very complex nature of metal corrosion which takes place during the oxidation of a lubricating oil at elevated tem- peratures, especially in the presence of organic sulfur compounds, one should recognize that totally different results might be expected when the same compounds are used with other bearing metal surfaces, or when experimental conditions are appreciably changed. Materials Employed Many of the sulfur compounds listed in Table II were obtained directly from vendors in a high degree of purity. This would include 3 Private communication, E. W. Fuller, Socony-Mobil Oil Co., Paulsboro New Jersey. Sulfur Compounds as Inhibitors in Oil Bearing Corrosion 25 such compounds as those numbered | to 8 incl., 10 to 15 incl., 22 and 32 in Table II. No. 6, however, contained a mixture of isomers. The sulfides numbered from 23 to 29 incl. were synthesized from alcoholic sodium sulfide and the necessary alkyl bromide in each case. No. 30 and No. 31 were prepared from ethylene oxide and the corre- sponding mercaptan. All of the disulfides listed in Table I, with the exception of compound No. 39, were made according to the method of Noller and Gordon (18) in which the alkyl bromide was allowed to re- act with an alcoholic solution of sodium disulfide. Tert-dodecy] disulfide (No. 39), however, was prepared from the sodium salt of tert-dodecyl mercaptan and iodine dissolved in CCl,. The sulfones, No. 16 to 21 incl., were prepared directly from the sulfides by oxidation with acid permanganate solution. The three sulfide polymers in the miscel- laneous group (Nos. 43, 45 and 46) were prepared from the necessary polymethylene dibromide in each case and alcoholic sodium sulfide solution. Ethylene disulfide polymer (No. 44) resulted from the reaction between ethylene bromide and sodium disulfide solution. 1,4-Dithiane (No. 47) was obtained as a by-product in the prepara- tion of ethylene sulfide polymer (No. 42) from ethylene bromide and sodium sulfide. Methyl-n-amyl-1,3-dithiane (No. 48) was the con- densation product from 1,3-propanedithiol and methyl-n-amyl ketone in the presence of dry hydrogen chloride. The blank oil used as a base in all of the corrosion tests was a solvent refined, S.A.E. 20 grade, paraffinic type of oil generously fur- nished for our purpose by the Socony-Mobil Oil Company. Some of the physical characteristics and analytical properties of this oil are listed below. Table 1.— Properties of Base Oil Rea ALPATE csc. coesesensnsees 81.0 WVascosity: Index >...) 5..208 110 Specific Gravity, 60°F. ........ 0.8708 Sulftir, (iat fs eae 0.08 IPYOUPS SO) De! Sosre eens ape earns 20 Refr. Index (D line TRG is SORE eee 450 DOSGA) ee aeecee eee eres 1.4801 Eunice seat ats eay cake tecesesreetccees: 490 Aniline Pomt, 1G, xccc.detes Je Kinematic Viscosity Waterman Ring Analysis Bip MIO? Oe escosccoseseocreosasa eee 66.99 Aromatic Rings, % ........ 5.9 Fi ALG (0 anna merle ee neieaes 32.76 Naphthenic Rings, % .... 19.7 GE OAC) 2) CUR te oo ee 8.69 Parraffinic Chains, % .... 74.4 Rings/Molecule ................ 2.0 Apparatus Details of an apparatus designed to measure the extent of bearing corrosion in a lubricating oil in the presence of air at 175°C. are shown in Figure 1. It is similar to the cadmium silver bearing corrosion test used in some lube oil laboratories and previously described in the pat- 26 N. W. Hall, W. D. Williams, and J. R. Meadow ent literature (15, 16). Except for a few modifications the conditions used in our work were very similar. Air at the rate of two liters per hour, controlled by the pressure regulator system at C in Fig. 1, was Fig. 1. Diagram of corrosion test apparatus. blown into a 25-gram sample of oil at H for a period of twenty-two hours. Temperature of the oil samples was kept at 175°C. by means of a well-insulated constant temperature bath (A), equipped with a stirrer, 250-watt heater, thermometer, and thermoregulator (+0.5°C.). Extra heavy lubricating oil served as a heat transfer medium. A sep- arate air supply for each sample entered the system at J, passed through a pressure regulator, D, a drying tube at L, thence to the test cell at E, and escaped through exit tube G and a glass jet at N. By means of a stopcock M the air current from the test cell E could be diverted through O to a flowmeter P. The standard test piece used was one quarter of a Pontiac connect- ing rod bearing, 4 x 1.4 cm., weighing approximately 6 grams and having a surface of cadmium silver alloy. A clean, weighed strip (I in Fig. 1) was removed after completion of a run, washed with petroleum ether, dried at 120° C., and reweighed. The loss in weight before and after a determination was reported in milligrams (see Table II). Experimental Results Under the experimental conditions noted above, the blank oil whose properties are recorded in Table I gave an average bearing weight loss of about 18 mg. when no inhibitor was added. Table II summarizes the results obtained with the various sulfur compounds Sulfur Compounds as Inhibitors in Oil Bearing Corrosion 27 TABLE Il” RESULTS OF THE CORROSION TEST ON OIL SAMPLES CONTAINING SULFUR COMPOUNDS AS INHIBITORS* Sulfur Compound Used Average Bearing Sulfur Compound Used Average Bearing as Inhibitor Weight Loss as Inhibitor Weight Loss in Mg. b in Mg.> BLANK OIL (No inhibitor added) 18,0 SULFIDES MERCAPTANS 22, n-Heptyl sulfide, B.p, 143-145°/8 mm, 21.8 1, n-Heptyl mercaptan, B.p. 174-6° 46.4 ; OL a i a 23, n-Octyl sulfide, B,p. 131-134°/1.8 mm, ‘21.7 2, n-Octyl mercaptan, B.p, 201-3° 10, 2 : a Es " M4 3 9 2h, 2-Octyl sulfide, B.p. 100-102°/1.7 mm, 8,5 3, mn-Nonyl mercaptan 23.0 25, n-Decyl sulfide, M.p. 24-25° 14,3 4, n-Undecyl mercaptan 5.9 26, n-Dodecyl sulfide, M.p. 38-40° 17.5 » NeDodecyl mercaptan, B.p. 146-8°/15 mm, 38 : 2 * See ie 3 2 27. n-Tetradecyl sulfide, M,p, 51-52° 11,6 6, tert-Dodecyl mercaptan, B.p, 78-87°/5 m, 0,2 rs ne Dla, 28. n-Hexadecyl eulfide, M.p, 54-56° 14 » mneTetradecyl mercaptan, B,p, 177-181°/21 mm; au 7 sa: OAs 3 29. n-Octadecyl sulfide, M.p. 66-68° 4,6 8. n-Octadecyl mercaptan, B.p; 212-4°/15 mm, 0.3 a 30. Phenyl £-hydroxy ethyl sulfide,M,p.56-58°77.7_ 9. Trimethylene dimercaptan, B.p, 66-8°/18 mm, 4, ‘ F ony ‘ gh 9 31, n-Amyl f-hydroxy ethyl sulfide,B.p. 125- THIOPHENOLS 126°/10 mm, 45.5 10, Thiophenol, B.p, 62-3°/15 mm, 57.2 32, Phenyl sulfide, B.p, 151-153°/15 m, 17.7 11, p-Thiocresol, M.p, 41-43° 58.6 DISULFIDES 12, o-Thiocresol, M.p, 10-12° 48,9 33, n-Butyl disulfide, B.p. 110-113°/15 mm, 26.5 13, m-Thiocresol, B.p. 75-7°/10 m, 65.5 34, n-Heptyl disulfide,B.p,198-210°/22 mm, 2,1 1h, p-Octadecyl thiophenol, Mp. 55°55.5° 0.2 © 35. n-Octyl disulfide,B,p.180-166°/5 mm, 5.9 15, £-Thionaphthol, M.p, 80-82° 45,1 36.°2-Octyl disulfide,B.p, 125-129°/1.7 mm, 4,5 37. n-Decyl disulfide, B.p, 15-16° 2.7 38, n-Dodecyl disulfide, M.p. 26-27°C 2.8 SULFONES 39, tert-Dodecyl disulfide, B.p. 60-90°/0.2- A = ee 0.3 m 0.3 16, n-Heptyl sulfone, M.p, 79-79,5° 23.3 . 40, n-Tetradecyl disulfide, M.p. 44,5-45° 2.0 17. n-Octyl sulfone, M.p. 75-76° 12.0 7 Fs : 41, n-Octadecyl disulfide, M.p. 48-48.5° One 18, n-Decyl sulfone, M.p. 83-84° 21.5 MISCELLANEOUS. 19, n-Dodecyl sulfone, Mp. 93.5-94° 20.6 42, Elemental sulfur 0,0 20, n-Tetradecyl sulfone, M.p. 98-98.5° 24.7 : 43, Ethylene sulfide polymer, M.r, 148- 21, n-Octadacyl sulfone, M.p. 105-106° 16.0 159° 0.4e 4h, Ethylene disulfide polymer,M.r.98-115° 0.8° 45, Hexamethylene sulfide polymer, M.r,54- bee 1.4° 46, Decamethylene sulfide polymer, M.r,71l- 76° 7.0 47, 1,4-Dithiane, M,p, 112° 21.4 48, Methyl-n-amyl-1,3-dithiane, B.p, 1h0-142/15 mm, 9.9 8£ach oil sample contained 0,2% sulfur from added inhibitor. é bresulting from the average of two or more determinations after running for 22 hours at 175° C. CAll temperatures reported are Centigrade readings, uncorrected, dcontained a mixture of isomers, Bearing showed a gain in weight, frhis sample contained only 0.1% sulfur instead of 0,2% as in others, Spoiling range not listed by vendor's catalogue (Columbia Organic Chemicals Co,), when they were used as inhibitors in amounts sufficient to give oil samples containing 0.2% sulfur on a weight basis. These quantities were equivalent to 0.0062 mole per 100-gram oil sample for compounds containing only one sulfur atom per mole, or 0.0031 mole for com- pounds with two sulfur atoms, etc. 28 N. W. Hall, W. D. Williams, and J. R. Meadow For some of those compounds which caused little or no bearing weight loss, as indicated by results in Table I, a few additional cor- rosion tests were made with oil containing smaller amounts of the sulfur inhibitor. These results are indicated in Table III, and as in the previous tests they represent the loss in weight, in milligrams, of the cadmium silver bearing strip after contact with a 25-gramm oil sample for 22 hours at 175°C. TABLE IIL CORROSION TESTS ON SOME OIL SAMPLES WITH REDUCED AMOUNTS OF SULFUR INHIBITORS* _———————————— eee eee b Bearing Weight losses in Mg. for different concentrations of inhibitor© Sulfur Inhibitor 0.2% 0.1% - 0.05% 0.037% 0.025% Ethylene sulfide polymer (No. 439) o.4& 0.2° 3.6 11.6 19.2 Hexamethylene sulfide polymer (no, 45) 1h 0.2 16.3 seen RES Decamethylene sulfide polymer (No. 46) 7.0 12.5 -oee ecee cans Ethylene disulfide polymer (No. }) onee 0.8 7.2 eooe onnn tert-Dodecyl mercaptan (No, 6) 0.2% 2h 1 cane ecoo mae p-Octadecyl thiophenol (No. 14) 0.0 0.2 eons cose ance Elemental sulfur (No. 42) 0.0 0.2 O.1 1.0 eens See aResults expressed as average weight loss in mg, for each different concentration of inhibitor used with conditions of time and temperature similar to those in Table II, badded to 25.g sample of base oil, °Expressed in terms of weight percent of elemental sulfur, dyumbers refer to compounds listed in Table I. s factually this represents a slight gain in weight of the bearing strip, Some oil inhibitors may appear to be satisfactory from the stand- point of preventing or reducing the amount of corrosion of a cadmium silver bearing surface for a limited time, but they may be quite un- desirable because they tarnish copper. Because of this, inhibitors are usually subjected to the copper strip test as a preliminary screening operation before other tests are actually carried out. A simple quali- tative test is to permit a small sample of oil to remain in contact with a strip of copper for 24 hours at 100°C. without agitation by air or stirring. Such a copper tarnish test was made on some of the better sulfur-containing inhibitors shown in Tables II and III. The results of the copper strip test are listed in Table IV, giving the concentration of inhibitor expressed in terms of percent sulfur, and the extent to which the added substance attacked the surface of the copper strip. Discussion A systematic study has been made of the corrosion inhibiting effect of several different kinds of organic sulfur compounds when added to oil samples in quantities to give 0.2% or less sulfur by weight. The compounds listed in Table II can be divided roughly into three types: (1) those compounds which had no effect; (2) those which Sulfur Compounds as Inhibitors in Oil Bearing Corrosion 29 TABLE IV EFFECT OF SULFUR-CONTAINING INHIBITORS ON COPPER STRIPS Per Cent Effect | Per Cent Effect Sulfur Inhibitor Sulfur in on Sulfur Inhibitor Sulfur in on E Oil Sample Copper - Oil Sample Copper MERCAPTANS : SULFIDES tert-Dodecyl mercaptan (No, 6°) 0.2 Severe” n-Tetradecyl sulfide (No, 27) 0.2 Extremely slight n-Tetradecyl mercaptan (No. 7) 0,2, Slight n-Hexadecyl sulfide (No. 28) 0.2 Extremely slight n-Octadecyl mercaptan (No, 8) 0.2 Considerable n-Octadecyl sulfide (No, 29) 0.2 Extremely slight DISULF IDES MISCELLANEOUS n-Heptyl disulfide (No, 3h) 0.2 Severe Elemental sulfur (No, 42) 0,05 Severe n-Octyl disulfide (No. 35) 0.2 Severa Ethyleme sulfide polymer (No. 43) 0.05 None 2-Octyl disulfide (No, 36) 0.2 Slight Ethylene disulfide polymer (No, 44) 0,1 Considerable n-Dodecyl disulfide (No, 38) 0,2 Considerable Hexamethylene sulfide polymer (No,45)0.1 Slight n-Tetradecyl disulfide (No, 40) 0,2 Slight Decenstip tore sulfide polymer (No.46)0.2 None n-Octadecyl disulfide (No, 41) 0.2 Severe p-Octadecylthiophenol (No. 1}) 0.2 None Numbers in parenthesis refer to compounds listed in Table Ir. bextent of tarnish on copper strip indicated in following order: None, extremely slight, slight, considerable, and severe. had a catalytic effect; ie. caused a weight loss of more than 18 mg,; and (3) those which produced an inhibiting effect on the oil. A few generalizations as follows can be made from this study: (1) The corrosion inhibition effect of a particular type of com- pound usually became more pronounced with an increase in molecular weight; this was more noticeable with the sulfides, the mercaptans and the disulfides than with some of the other sulfur derivatives em- ployed (see Figures 2-A and 2-B). (2) All compounds tested which contained the octadecyl radical, with the possible exception of the sulfone, were quite effective as inhibitors of corrosion on cadmium silver surfaces; this was not neces- sarily true on copper, however. (3) The mercaptans and disulfides which contained twelve or more carbon atoms per mole were found to be surprisingly good in- hibitors of corrosion on cadmium silver but unfortunately they tar- nished copper surfaces very badly. (4) Our work, in general, supports some of the observations of Denison and Condit (7), namely that alkyl sulfides as a group are probably the best all-around inhibitors among the sulfur compounds tested; this is especially true if one includes the oil-soluble polymeric sulfides, such as ethylene sulfide polymer (No. 43 in Table II). Figures 2-A and 2-B show the comparative inhibiting effect on cadmium silver surfaces of straight chain aliphatic sulfides, mercap- tans and disulfides when the chain length, and hence the molecular weight, is increased. The polymethylene sulfide and disulfide polymers were not included in the graph. Those compounds which caused a 380 N. W. Hall, W. D. Williams, and J. R. Meadow MILLIGRAM WEIGHT LOSS OF CADMIUM SILVER BEARING STRIP MTS Sy 8 HY ao sh ws a BBG mag ae NUMBER OF CARBON ATOMS IN ALKYL CHAIN Fig. 2. Comparative inhibiting effect of straight chain sulfur compounds. A—Alkyl sul- fides; B—Alkyl mercaptans and disulfides. bearing loss above 18 mg. might be considered as catalysts for bear- ing corrosion. Figure 3 gives the corrosion rate curves from 0 to 22 hours for the straight chain aliphatic mercaptans. It is admitted that results might vary somewhat with different stocks of base oil used, but due to its low sulfur content it doubtless served our purpose very well. It can be observed that the alkyl sulfides containing over twenty carbons per mole, the mercaptains and disulfides with at least twelve carbons per mole, the polymeric sulfur compounds, and a single thiophenol with an octadecyl group were fairly effective as inhibitors on cadmium silver. The sharp increase in the inhibiting effect of the higher molecular weight mercaptans was unexpected. Sulfur Compounds as Inhibitors in Oil Bearing Corrosion 31 SILVER BEARING STRIPS MILLIGRAM WEIGHT LOSS OF CADMIUM- Oar: ST eas 8 12 16 20 24 TIME IN HOURS Fig. 3. Rate of corrosion curves for straight chain aliphatic mercaptans. The alkyl sulfones were generally ineffective either as inhibitors or as catalysts for this type of corrosion test. This observation is in agreement with some of the results reported by Denson and Con- dit’ (7). Although some results were obtainable for studying the effct of chain branching on inhibitor action in this particular base oil, one must exercise caution in making conclusions from the limited data which are available. There is some indication that branched alkyl chains were slightly more effective than the corresponding straight chain isomers. Compounds listed in Table III were added to oil samples in quan- tities to give less than 0.2% sulfur. It can be observed that elemental sulfur was very effective at concentrations as low as 0.037%, and ethylene sulfide polymer (No. 43) was quite effective at 0.05% or even lower. p-Octadecylthiophenol (No. 14) gave favorable results at 0.1% concentration; unfortunately lower concentrations of this com- pound were not tried during the period of experimentation. De- camethylene sulfide polymer (No. 46) and tert-dodecyl mercaptan (No. 6) were not very effective when reduced from 0.2% to 0.1%. Hexamethylene sulfide polymer (No. 45) and ethylene disulfide poly- mer (No. 44) were effective at 0.1% but much less so at 0.50% con- centration. If compounds classed as inhibitors based on the Cd-Ag corrosion test are ruled out because they tarnish copper (copper strip test, Table IV), the number of satisfactory inhibitors listed in Table II 32 N. W. Hall, W. D. Williams, and J. R. Meadow would be reduced considerably. In fact, this would eliminate all but the alkyl sulfides, the polymethylene sulfide polymers, and p-octade- cylthiophenol. The latter compound probably deserves more study as an oxidation inhibitor for oils and is a type of compound which was mentioned in the work of Reid and Hamilton (16). The mechanism of corrosion inhibition may be due to any one or a combination of various factors. The most probable are the following: (1) Formation of a protective coating on the bearing surface.* (2) Reduction of acid formation in the oil by furnishing basicity. (3) Retard oil oxidation possibly by preventing the formation of peroxides. (4) Reduction of peroxides, i.e. destroy the peroxides formed. It has been demonstrated that the attack of oxygen on a hydro- carbon is a free radical chain reaction (19, 20). Denison (6, 7) has suggested that corrosion is a result of the ability of peroxides to con- vert metal into metal oxides which are subsequently removed by reaction with acidic constituents developed during oxidation. By a combination of these concepts, the mechanism of bearing corrosion may be illustrated by the following series of reactions: A. Formation of peroxides through chain reaction RH (activation) — R. + H. R. + O2 — ROO. ROO. -+- RH — ROOH -+ R. B. Corrosive action ROOH + M — ROH + MO MO + 2HA — MA, + H:O where M = Metal, ROOH = peroxide, and HA = organic acid. Several authors (7, 21) have suggested that the best corrosion inhibitors are probably those which readily reduce hydroperoxides. Thus, a sulfide inhibitor might destroy a peroxide by reduction as follows: ROOH + R-S-R — R-SO-R + ROH ROOH + R-SO-R > R-SO.-R + ROH The oxidation products of the sulfide might be either a sulfoxide or a sulfone. Our work has demonstrated that the sulfones as a group are quite inert as corrosion inhibitors for cadmium silver surfaces (see Table 4 This might explain the slight increase in bearing weight noticed in a few instances and recorded in Table II. Sulfur Compounds as Inhibitors in Oil Bearing Corrosion 33 Il). This particular observation of ours lends much support to the Denison-Condit mechanism described above. Summary Alkyl sulfides, disulfides and mercaptans were more effective as oxidation inhibitors than sulfones as determined by the cadmium sil- ver bearing corrosion test. Their effectiveness as inhibitors increased with molecular weight. Polymethylene sulfide polymers were most effective. With the exception of p-octadecylthiophenol, the thio- phenols appeared to be more corrosive than the mercaptans. The tar- nish effect on copper increased in the following order: polymethylene sulfide polymers, alkyl sulfides, alkyl disulfides and mercaptans. Some of our results lend support to the Denison-Condit idea of peroxide reduction in oils. Acknowledgment The authors wish to thank E. W. Fuller of the Research and De- velopment Laboratories, Socony-Mobil Oil Company, for his valuable suggestions relating to corrosion tests, and also for the base oil which he supplied for this work. Literature Cited 1. Weiland, W. F., 1950. Sci. Mo., 71:121. Larsen, H. G., Armfield, F. A., and Whitney, G. M., 1943. S.A.E. Jour., 51:310. 8. Zuidema, H. H., 1946. Chem. Rev., 38:197. ° 4. Byers, J. H., 1936. Natl. Petroleum News, 28:78-84 (No. 51). 5. Davis, L. L., Lincoln, B. H., Byrkit, G. D., and Jones, W. A., 1941. Ind. and Eng. Chem. 33:339. 6. Denison, G. H., Jr., 1944. Ind. and Eng. Chem. 36:477. 7. Denison, G. H., Jr., and Condit, P. C., 1945. Ind. and Eng. Chem. 37:1102. 8 9 bo . Dornte, R. W., 1936. Ind. and Eng. Chem. 28:26. . Fenske, M. R., Stevenson, C. E., Lawson, N. D., Herbolsheimer, G., and Koch, E. F., 1941. Ind. and Eng. Chem. 33:516. 10. Haslam, R. T., and Frolich, P. K., 1927. Ind. and Eng. Chem. 19:292. 11. Lamb, G. G., Loane, C. M., and Gaynor, J. W., 1941. Ind. and Eng. Chem., Anal. Ed. 18:317. 12. Larsen, R. G., Armfield, F. A., and Whitney, G. M., 1943. S.A.E. Journal 51:810. 13. Larson, R. G., Thorpe, R. E., and Armfield, F. A., 1942. Ind. and Eng. Chem. 84:1838. 14. Loane, C. M., and Gaynor, J. W., 1945. Ind. and Eng. Chem., Anal. Ed. 17:89. 15. Mead, B., 1927. Ind. and Eng. Chem. 19:1240. 16. Reid, E. E., and Hamilton, L. A., 1941. U.S. Patent No. 2, 230, 966. 34 N. W. Hall, W. D. Williams, and J. R. Meadow 17. Moran, R. C., Evers, W. L., and Fuller, E. W., 1936. U.S. Patent No. 2, 058, 342. 18. Noller, C., and Gordon, J., 1933. J. Am. Chem. Soc. 55:1090. 19. Frank, C. E., 1950. Chem. Rev., 46:155. 20. Waters, W. A., 1948. “Chemistry of Free Radicals”, 2nd Ed., Oxford Press, London, England. (p. 232-234). 21. Gruse, W. A., and Stevens, D. R., 1942. “Chemical Technology of Petroleum”, McGraw-Hill Book Company, New York. Accepted for publication January 4, 1960. THE CAVE SNAIL, CARYCHIUM STYGIUM CALL LESLIE HUBRICHT 1285 Willow Avenue, Louisville 4, Kentucky Carychium stygium Call, was described from specimens collected in Mammoth Dome, Mammoth Cave, Edmonson County, Kentucky. (Call, 1897) Giovannoli (Bailey, 1933) found it in Buzzard’s Cave and White’s Cave, near Mammoth Cave. This remained the known range of the species until the present author found it to be widely distributed in the caves of central Kentucky. It was found in an area about 60 miles long, and about half as wide, extending from near Upton, Kentucky on the north to near Portland, Tennessee on the south. In this area it was found in most of the caves in which there was sufficient food and moisture. Efforts to find it outside of caves were unsuccessful; it was found only in the total darkness of caves. ——————— Fig. 1. Carychium stygium Call. Whites Cave. A. head of animal with round eyes and rounded tentacles. B. head of animal with degenerate eyes and pointed tentacles. C. three shells opened to show lamellae. Scale line equals 1 mm. Localities: KENTUCKY: Hart County: Copelin Cave, 2 miles east of Millerstown; Puckett Cave, 1 mile west-southwest of Priceville; Chattin Cave, 2 miles west of Priceville; Cooch Webb Cave, 2.2 miles west of Priceville; Buckner Hollow Cave, 7 miles east-northeast of Munfordville (Thomas C. Barr, Jr., Coll. ); Cub Run Cave, 2 miles west of Cub Run; Ronalds Cave, 2.6 miles north of Cave City; Hogan Cave, 3 miles north of Cave City; cave, 2 miles southwest of Northtown. Edmon- son County: near river in Great Onxy Cave; Cathedral (Buzzards) Cave, near Floyd Collins Crystal Cave. Mammoth Cave National Park: Pogoda Cave; Salts 36 Leslie Hubricht Cave, near old Pike Chapman Entrance; Whites Cave; Little Whites Cave; Dixon Cave; Proctors Cave; Martins Cave; Running Branch Cave; Blowing Spring Cave; small cave near Longs Cave; Mammoth Cave: Bunker Hill, end of Audubons Ave- nue; Mammoth Dome; near Richardson Spring; River Hall; Violet City; Cathedral Domes; New Entrance; Frozen Niagara. Barren County: Indian Cave, 4 miles west of Cave City; Railroad Cave, Cave City; Burnett Cave, 0.6 mile west of Park City; Vance Cave, 0.8 mile northwest of Park City; Brushy Knob Cave, 2 miles northwest of Park City; Diamond Cavern, 2 miles north of Park City (fossil only); Short Cave, 2.2 miles northwest of Park City; Beckton Cave, 0.5 mile northwest of Beckton; Duval Saltpeter Cave, 0.7 mile northwest of Beckton; Cave Spring Cave, 1 mile south-southwest of Red Cross. Warren County: Bypass Cave, Bowl- ing Green; Vails Cave, 2 miles west of Bowling Green. Simpson County: Hoy Cave, 2 miles north of Franklin; Steeles Cave, 4 miles southeast of Franklin. TENNESSEE: Sumner County: small cave in sink above White Oak Cave, 2.2 miles east-northeast of Mitchellville. Considering the isolation of the colonies, there is remarkably little variation. In shell size, the over-all variation is only slightly greater than that to be found in a single colony. (Fig. 2). The largest shells were found in Mammoth Cave and other caves in the vicinity. They become slightly smaller at the northern and southern limits of the range. In the shells opened there was very little variation in the internal lamellae. The principal lamella has an even spiral edge and is rather small. The columellar lamella is obsolete (Fig. 1C). In the animals examined the tentacles were either bluntly pointed (Fig. 1B) or lobed (Fig. 1A), the latter being in the majority. The eyes are situated in the base of the tentacles and are usually round (Fig. 1A), but sometimes are reduced to an irregular mass of dark pigment (Fig. 1B). The question of how Carychium stygium became distributed in the caves in which it is found is as yet unanswered. That this snail, whose movement is so slow as to be barely perceptible, could crawl from a single point of origin through subterranean passageways into the many caves in which it is now found seems improbable. It seems more probable that it was originally a surface species which moved inde- pently into each of the cave systems in which it is now found. The absence of variation and the presence of eyes would indicate that either the migration into the caves has been rather recent or that the snails have an unusually stable gene system. Much has been written about the uniformity of the cave habitat. While it is true that temperatures and the absence of light are con- stant, other important factors, such as moisture, food supply, and the prevalence of competitors and predators, may vary widely. Some caves are quite wet, with dripping walls and ceilings, often with stalactites and stalagmites. These usually have an abundant fauna if food is available. Others are subject to flooding after heavy rains; these caves usually do not have as abundant terrestrial fauna as the first type. The Cave Snail, Carychium Stygium Call AAA AAABAE seaicaiss ddddddade PAA AA AAA ee Fig. 2. Carychium stygium Call. A. Copelin Cave. B. Cooch Webb Cave. C. Buckner Hollow Cave. D. Cub Run Cave. E. Ronalds Cave. F. Hogans ave. G. Mammoth Dome, Mammoth Cave. H. River Hall, Mammoth Cave. |. Brushy Knob Cave. J. Duval Saltpeter Cave. K. Bypass Cave. L. Steeles Cave. Scale line equals 1 mm. Still others may be relatively dry, and usually have a meager fauna. There are two principal sources of food supply; debris washed in by streams; and the guano of bats, cave rats (Neotoma), and cave crickets (Hadenoecus). My observations indicate that while Carychium stygium will feed on leaves and wood where it is available, the principal food is the 38 Leslie Hubricht guano of the cave cricket (Hadenoecus subterraneus Schudder ). They have not been observed feeding on either bat or cave rat guano. In most caves there is a direct correlation between the number of cave crickets and the abundance of Carychium stygium. Since the cave crickets usually must go outside of the caves at night to feed, they are not found in abundance very far from an opening. Carychium stygium is usually found near the entrance (but in total darkness), or in the vicinity of breakdowns. In Mammoth Cave they are found in the deeper parts of the cave only in River Hall, where they appear to feed on mud and slime brought in by the annual floods. Usually associated with Carychium stygium, in addition to Hade- noecus subterraneus, are the cave millipede Scoterapes copei (Pack- ard), the bristle-tail Campodea cookei Packard, and the beetle Ptoma- phagus hirtus (Tellkampf). These are guano feeders and compete with Carychium stygium for food. The harvestman Phalangodes ar- mate Tellkampf, the spider Antrobia mammouthia Tellkampf, and the beetles Neophaenops tellkampfi (Erichson), and Pseudanophthalmus spp. are predators. Whether any of these prey upon Carychium sty- gium is doubtful. None was seen to show any interest in the snails. Carychium stygium requires a very moist habitat and drought is fatal. On the other hand, it seems to be able to stand submergence in water for long periods. It was found crawling about on the wet mud after a flood in River Hall, Mammoth Cave. In White’s Cave it was seen crawling about on the bottom of a pool where there had been water for several months. Literature Cited Call, R. E., 1897. Some notes on the flora and fauna of Mammoth Cave, Ken- tucky. Amer. Nat. 31: 377-392. Bailey, Vernon, 1933. Cave Life in Kentucky. Amer. Midl. Nat. 14: 385-636. APPLICATIONS OF SOLAR ENERGY K. V. PRASANNA Illinois Institute of Technology, Chicago, Illinois The amount of energy radiated at the sun’s surface is about twenty million B.T.U. per square foot per hour. Of this, the energy received at the outer edge of the earth’s atmosphere amounts approximately to two thousandths of one per cent (0.002%). A maximum of forty percent of this energy intercepted could be collected at the earth’s surface, the remainder being absorbed and scattered in the atmos- phere. All the deposits of coal and oil comprise a very small fraction of this energy stored over millions of years. Water situated at high potential is another form of the sun’s energy. Fuel alcohol obtained from certain vegetable matter is yet another form of stored energy. If all the stored energy that could be recovered from the earth were stock piled, it would amount only to a three-day period of solar energy intercepted by the earth. It has been estimated that the energy consumed during the past fifty years corresponds nearly to eighty percent of all the energy ever consumed by Man. According to very reasonable estimates of the future power requirements, all the present energy, including atomic energy, will last for only 245 years. By 2200 A.D. Man must be able to harness one percent of the solar energy re- ceived by the earth to cope with his power requirements. Even the most modern equipment operating on present fuels cor- responds to sixteen hundredth of one percent of the energy received. It is needless to mention the efficiency of the equipment used by our predecessors. Let us consider the familiar example of the domestic animal the bull, which is still a chief source of motive power used by the agriculturists of India. An average Indian bull can exert a force of sixteen hundredth (0.16) of a ton for eight hours, the power of the bull being 720 watts for that period. The average power utilized is only a third of this, that is 240 watts for eight hour period. The aver- age for a twenty-four hour day is only eighty (80) watts per hour. On the other hand, the average minimum consumption of food is nine- teen pounds of dry hay per day, a heat equivalent of 1500 watts. A steam engine operating on the same fuel can develop one hundred and eighty (180) watts. These figures show how inefficiently we are utilizing our resources. If a power plant were to be built utilizing the solar energy re- ceived on 750 square miles of desert land in the United States, it could cater to the present power requirements. A power plant covering one- 40 K. V. Prasanna fifth of the area of the state of New Mexico can supply thirty times the present power requirements of the United States. If the harnessing of solar energy is broken into its elements, it falls into four branches: collection, transportation, storage and utilization. Let us consider the influence of each of these factors. The amount of solar energy received on any part of the earth’s surface depends on: (1) the hour angle of the sun (2) the declination (3) the latitude of the place (4) the solar altitude above the horizon (5) the azimuth of the sun . (6) atmospheric conditions like air mass factor, cloud, and haze factor, diffuse radiation factor (7) the position of the energy receiving surface. It is observed that, at the outer edge of the earth’s atmosphere a maximum of 4300 B.T.U. per square foot of normal area for a day of ten hour sunshine, can be received. The average amount of solar energy at any place is given by 4300 multiplied by all the above said factors. According to U.S. Weather Bureau reports, near the Boston area, minimum average incident solar energy is 1000 B.T.U. per square foot of south wall area per day. This figure is greater as we go towards the equator. The threshold value of incident solar energy below which it cannot be collected is thirty B.T.U. per square foot per hour which corresponds to 4.3% of daily total energy received. The amount of solar energy that could be collected depends on the efficiency of collectors and the temperature of collecting surface above that of the environment. The type of collector depends on the par- ticular application. For water and space heating, a flat plate collector can be used in which a temperature as high as 195°F has been at- tained. For conversion of solar energy into power a higher tempera- ture difference is necessary. For this a mirror type collector is used in which a temperature as high as 8500°F has been attained. How- ever, to keep the radiation losses to a minimum, a smaller temperature difference is advisable. A blackened flat copper plate, when used as a collector, has an absorption efficiency of ninety-five percent, but external radiation losses bring down the net collection efficiency to a low value. Intro- duction of a glass cover plate reduces the incident solar energy by a small amount, but it reduces radiation losses to a great extent, thus increasing the collector efficiency. Experimental investigation has shown that up to a maximum of three glass cover plates with air Applications of Solar Energy 41 spaces in between, improves collector efficiency considerably, the value of the efficiency in this case being fifty-seven percent. For concentration of solar energy by mirrors a concave or a para- bolic mirror can be used. However, from the cost consideration it is economical to build the collector using pieces of flat mirrors with proper orientation. It is needless to say that the transportation and storage losses should be kept at a minimum by the use of proper insulation so that overall efficiency of solar equipment can be kept at a high value. Utilization of Solar Energy The term utilization factor is defined as the ratio of energy avail- able in the required form, to the amount of solar energy incident on the collector. Experimental investigations have shown that utilization of solar energy as heat, is far more efficient than other forms of energy like mechanical and electrical energy, since the latter usually involves heat as an intermediate stage. The utilization factor is as low as five tenths of one percent (.5%) in photonynthesis. It has reached a peak of thirty-two percent (32%) for space heating, which figure is con- siderably greater than that obtainable from other forms of energy whose ultimate source is solar energy. Egyptians were one of the pioneers in utilizing solar energy for heating water and operating irrigation pumps. In the year 1818, the first steam engine operated completely by solar energy was built in France. It produced one horse power utilizing twenty square yards of collector area. A modified solar steam engine was built in 1878 which operated a complete printing press. In 1868, a solar furnace for melting copper was built. A solar steam engine built at South Pasadena, California operated a water pump, with a discharge capac- ity of 1000 gallons per minute. In more recent years several devices harnessing solar energy have been built all over the world. In Algiers a solar furnace equipped with twenty-seven and a half foot diameter parabolic mirror, synthesizes nitric acid directly from air, water, chalk and sunshine. In Pyrnees, France a forty foot mirror produces refractory ceramics such as fused quartz and titanium oxide on a commercial scale. Convair’s at San Diego, have built a solar furnace using one fourth inch aluminum plate bent into parabolic shape. The ten foot diameter mirror focuses the sun’s rays to a hot spot five sixteenths of an inch in diameter at thirty-four inches from the center of the mirror. The temperature attained is 8500°F as compared with a maximum temperature of 5800°F attained by oxy-acetylene torch. Dr. C. F. 42, K. V. Prasanna Kettering has developed a photo-electric cell converting directly light into D.C. current which runs a small electric motor. Solar batteries developed by Bell laboratories, use ultra-pure silicon strips one inch by four inches (1” x 4”) impregnated with one ten thousandth of an inch (0.0001”) boron. It is between these two surfaces transistor action takes place which results in flow of current. When several such elements are electrically connected, the battery is capable of giving a high current at a high voltage. The utilization factor for these bat- teries is eight percent. Now these batteries are commercially used for battery charging and in telephone circuits. The National Physical Laboratory of New Delhi, India has built an engine completely oper- ated by solar energy. The same laboratory has built a solar cooker using a mirror type collector. Another type of cooker that can be used indoors uses heat absorbed by the fluid. Similar cookers have been developed at M.I.T., Massachusetts, and Japan. According to Russian claims, giant reflectors operate textile factories; high pressure solar heaters cook fruits and vegetables in canneries; distill water, make ice and heat laboratories. Yet another application is the use of solar energy for hot-water heating. Hot-water heaters have been success- fully built both in California and Florida. A temperature as high as 140°F has been attained. Solar Energy for Space Heating One of the most feasible and economical application of solar energy for the present day, is its utilization for home heating. Experimental houses have been built in several parts of the United States to get an idea as to the various factors that influence space heating. The U.S. Weather Bureau over several parts of the country is regularly collect- ing data as to the amount of energy received by the earth. Maximum energy can be collected when the collectors are normal to the angle of incidence of the sun’s rays. It is almost impossible to achieve in practice since the angle of incidence is always changing. The best possible location of the collector for winter months, is at the south facing the roof inclined at sixty degrees to the horizontal. This has a disadvantage because of the accumulation of snow. To overcome the above disadvantage, collectors can be located at the south facing the vertical wall. This reduces the incident energy by ten percent but has the added advantage of collecting reflected solar energy during bright snowy days. Purdue University at Lafayette, Indiana, was the first to build an experimental solar house to investigate the possibility of using solar energy for space heating. The house built had the upper two thirds Applications of Solar Energy 43 of the entire south wall covered with glass. The roof overhang was so adjusted that it allowed maximum exposure to sunlight during winter and completely cut-off the sunlight during summer. The experimental facts showed that excess loss through the added glass area during the dark period was more than the energy gained during bright sunny day. The drawbacks were: (1) not covering the glass area by an insulated partition during the period when there was no solar energy gain, (2) not providing a means of storage when the energy received was in excess of the requirement. Dr. Maria Telkes at M.I.T. made an exhaustive investigation of several methods of storing systems with a view to building a success- ful solar house. The chief properties used were specific heat and latent heat of substances. Of all the substances, water has maximum specific heat, which permits maximum storage of heat for a given weight. On equal volume basis, it has a slight advantage since its specific weight is low. Experiments showed that due to excessive heat losses, it is not economical to store heat at a temperature higher than 100°F. Latent heats of melting of several chemical salts which lie in the range of 80 to 100°F can be used to a great advantage since a large amount of heat can be stored at constant temperature. One such salt is sodium sulphate, or commercially known as salt cake. The melting point of sodium sulphate is 90°F and the latent heat is 104 B.T.U. per pound. At the melting point most of the sodium sulphate melts in its own water of crystalization leaving only a small part of anhydrous salt. For instance, between 80°F and 100°F sodium sulphate can store 11,000 B.T.U. per cubic foot. To store the same amount of heat eight and a half cubic foot of water is necessary, which means larger space requirements. The impure sodium sulphate is quite cheap and cost of installation compares favorably with other types of specific heat storage. Possibility of storing a large quantity of heat at a constant temperature is of very great advantage for space heating. Inner cor- rosion can be prevented by the use of corrosion resistant materials. The heat receiving coil can be designed to prevent the settling of solid to the bottom by providing temperature stratification. Catalyst may be added to promote melting and recrystalization. The slight increase in volume during melting may be taken care of by partially filling the container. Salt packed in small containers is more economical and advantageous. Figure 1 shows the cross section of a house having a large south wall glass area which almost corresponds to one built at Purdue Uni- versity. In the vicinity of the Boston area during the winter months, on an average, the incident solar energy is 1000 B.T.U. per square foot 44 K. V. Prasanna DOUBLE GLASS With AIR SPACE TRANSMITTED - 720 NET GAIN - 220 Loss - $00 LFFICIENCY - 22 Y, figures ARE IN BTU PeR S0.Fr PER DAY Pic. 1 -So.rae Hous&é Witw LARGE Winoows of south vertical wall area per day. Energy transmitted through the double glass wall with air space is 720 B.T.U. per square foot per day. The outward heat loss is 500 B.T.U. per square foot per day leaving a net gain of 220 B.T.U. per square foot per day. On clear days there will be more gain of energy, overheating the house, and on cloudy days there will be more external loss which necessitates auxiliary heating. Figure 2 shows a similar house equipped with a chemical heat storage, using sodium sulphate as the chemical. The house is equipped with an insulating partition which is lowered during nights and dull days, reducing outward losses. The insulating partition at the inner end controls the room temperature at 70°F. For the same incident conditions the outward loss is 400 B.T.U. per square foot per day, and the net gain is 320 B.T.U. per square foot per day. Eight and a half inches represents the width of storage wall necessary to heat a four room house, the entire south wall being covered by the collector. By the figure it is apparent that this arrangement has the advantage of controlling the room temperature, added to increased utilization effi- ciency attained. Figure 3 indicates improvement over the ordinary type of storage wall. The insulation wall in between collector and storage reduces the outward loss. The hot air circulation system ensures a better way of transmission of energy absorbed. The outward loss in this case is reduced to 350 B.T.U. per square foot per day ee the net gain to 370 B.T.U. per square foot per day. Applications of Solar Energy 45 Utilizing these principles of storage systems, M.I.T. built a solar house which was completely heated by solar energy. The excess heat collected during clear winter days was stored in large storage bins to provide heat for intermediate spans of cloudy days when there was no gain of solar energy. Experimental investigations showed that it is economical to build storage bins as an integral part of the house, yeh GLASS W'TH AIR SPACE INSULATING PARTITION LOWERED git Ar NicHT f i | ONTROL 4 . Room TEMP. - TOF Sun-1000 OUTWARD Loss-400 NET GAIN EFFICIENCY -32 7, Fisures ARE IN BTU FER Sa. fr FER Day 276.2 - SUN Want Wirn CHemical HEAT STORAGE CHEMICAL ~ GLAUBER'S SALT (NazS0s 1020) ie DouBLeE GLASS WITH AIR SPACE Air Duct _»- HEAT CONTROL / Sun 1000 Room TEMP. ~ TOF ColtEcror TEMP. Less THaAn 110°F OUTWARD | EFFICIENCY-37/, STK) INSULATED FAN, ix Bs Fi00R pel a EEE Fisures ARE INBTU PER Sa.Fr. PER Day FIG. 3-WALLTYPE CHEMICAL HEAT STORAGE CHEMICAL~ GLAUBER'SSALT (NazS04 10 4,0) 46 K. V. Prasanna situated in between two rooms to be heated. With this the radiation losses of the bins can be used to heat the rooms, raising the utilization efficiency. It was felt that, to equip the house with a complete solar heating system, to take care of long, infrequent, no sunshine days, necessitates the building of large storage bins. To overcome this dis- advantage, it was felt that the house should be provided with any of the present day equipment as an auxiliary unit. Several advantages can be attained by the installation of an auxil- iary heat pump using air water or earth as a heat source, along with solar collectors. Provision of a higher heat source improves the effi- ciency of the heat pump. The temperature of collection need not be as high as any of the previously described systems since the heat pump acts as a booster and raises the collector’s efficiency. The excess heat collected can be stored in the earth (in the case of an earth heat pump), saving the cost of storage equipment. There is no necessity to build the collectors as an integral part of the house. During sum- mer the solar collectors can be used for hot water heating and the heat pump can be operated on the cooling cycle for summer air-condition- ing. In the future, if all the American homes were equipped with solar heating systems, the energy saved would be more than fifty per- cent of the total American fuel consumption. Today it is not impractical to think of a solar engine even up to a capacity of five horsepower which is quite a handy piece of equipment in an American farm home. Figure 4 shows a typical solar engine that can be built for such purposes. The solid line shows a freon-12 cycle and the dotted line shows a water cycle. The water cycle is equipped MiRROR CONCENTRATION Ve ON SUPERHEATER FREON-12 EVAPORATOR FUATPLATEABSORBER 4 (irs WATER CIRCULATION |} bm <--— pe aa CONDENSER FREON -12 —--— WATER Lie. 4 - TYPICAL SOLAR ENGINE Applications of Solar Energy 47 with a pump to circulate, and a flat plate absorber to heat the water. The hot water passes through a heat exchanger evaporating freon in the evaporator. The freon vaper passes through a super heater equipped with mirror concentration. The super-heated vapor expands through an engine generating useful power. The exhaust vapor passes through a condenser and a circulating pump back to the evaporator. Another possible application is the use of solar energy for summer air conditioning. Figure 5 shows a typical solar air-conditioner. At- HEATER COOLER TURBINE” N\ Compressor EXPANSION TURBINE EXHAUST Air From Colo AIR FOR ATMOSPHERE AIR-CONDITIONING PIG6.5 ~TyPICAL SOLAR AlR-CONDITIONER mospheric air is drawn into a compressor and compressed to a high pressure. A small part of this high pressure air passes through a solar heater equipped with mirror concentration. The air at high pressure and temperature, passes through a turbine, which provides motive power for the compressor. The major part passes through a conven- tional water cooler. The cool high pressure air passes through an expansion turbine, with a further drop in temperature. The expansion turbine provides additional power for the compressor. The cold air coming out of the expansion turbine is available for air conditioning and human comfort. The ever increasing power requirements of the modern man and the limited power resources at his disposal will certainly provide a great incentive to the already curious scientific investigator to explore successfully the possibility of harnessing the everlasting sun’s energy for his power requirements. Before long, the economical barrier that is still enshrouding the development will break down and we will see man utilizing solar energy for his power requirements, not to mention the small innumerable household appliances. Very soon historians will 48 K. V. Prasanna have to change the statement that, the prosperity of a nation depends on its coal deposits, to the prosperity of a nation depends on the extent to which it can harness the mighty and inexhaustible solar energy. Bibliography 1. Hutchinson, F. W., “Solar House: A Full Scale Experimental Study,” Heating and Ventilating, Vol. 42, No. 9, September ’45, Page 96. Hutchinson, F. W., “Solar House: Research Progress Report,” Heating and Ventilating, Vol. 43, No. 8, March ’46, Page 53. 3. Hutchinson, F. W., “Solar House: A Second Research Progress Report,” Heat- ing and Ventilating, Vol 44, No. 3, March ’47, Page 55. 4. Telkes, Maria, “Solar House Heating: A Problem of Heat Storage,” Heating and Ventilating, Vol. 44, No. 5, May ’47, Page 68. 5. Telkes, Maria, “A Review of Solar House Heating,” Heating and Ventilating, Vol. 46, No. 9, September 749, Page 68. 6. “Utilization of Solar Energy,” Engineering, Vol. 174, Nov. 7, 1952, Pages 607-8, Engineering, Vol. 174, Nov. 14, 1952, Pages 656-6, Engineering, Vol. 174, Nov. 21, 1952, Page 679. 7. Heywood, H., “Solar Energy: Past, Present and Future Applications,” Engi- neering, Vol. 176, Sept. 18, 1953, Pages 377-80, Engineering, Vol. 176, Sept. 25, 1953, Pages 409-11. 8. Jordan, R. C., and Threlkeld, J. L., “Design and Economics of Solar Energy Heat Pump,” Heating Piping and Air Conditioning, ASHVE Journal Section, Vol. 26, No. 2, Feb. ’54, Pages 122-30. 9. “Bell Makes Battery Powered by Sun,” Electrical World News, Vol. 141, May 10, 1954, Pages 47-8. bo ACADEMY AFFAIRS The fall meeting will be held at the University of Louisville on November 4 and 5. Titles of papers to be presented at the sectional meetings will be accepted by the section secretaries at any time prior to mid-October. INSTRUCTIONS FOR CONTRIBUTORS The TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE is a medium of | publication for original investigations in science. Also as the official organ of the Kentucky Academy of Science, news and announcements of interest to the member- ship are published therein. These include programs of meetings, titles, abstracts of papers presented at meetings, and condensations of reports by the Academy’s officers and committees. Papers may be submitted at any time to the editor. 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Telephone JU 3-0666 Telephone DI 3-5515 Volume 21 1960 Numbers 3-4 RANSACTIONS - BR EINTUCKY ACADEMY of SCIENCE Official Organ KeNTucKy ACADEMY OF SCIENCE CONTENTS Soil Temperature Measurements at Lexington, Kentucky, from 1952 to 1956 E. B. Penrop, J. M. Exxiott, and W. K. Brown Cyclic Entry-Exit for Subroutines J. C. Eaves and T. J. PicNANI The Salting Effect of Sodium Chloride on the Extraction of Erbium Acetylacetonate and 8-quinolinol Y. G. Isoipa, J. F. Srernspacu, and W. F. WAGNER A North American Leafhopper Previously Confused with Typhlocyba andromache McAtee (Homoptera, Cicadellidae ) Pau. J. CHRISTIAN Studies of Overwintering Potential of Certain Drosophila Species WALLACE D. Dawson Academy Affairs Index to Volume 21 The Kentucky Academy of Science Founded May 8, 1914 OFFICERS 1960-61 . President: H. H. LaFuzer, Eastern State College President-elect: CHARLES WHITTLE, Western State College Vice President: LyL= Dawson, University of Kentucky Secretary: Grennir LEvrEy, Berea College Treasurer: RicHarp A. CHAPMAN, University of Kentucky Representative to AAAS Council: Mary E. WHarton, Georgetown College Counselor to Junior Academy: Maurice CuristorHER, Murray State College OFFICERS OF SECTIONS BACTERIOLOGY AND MEDICAL TECHNOLOGY Chairman: GENEVIEVE CLark, Georgetown College Secretary: MARGARET Hotcukiss, University of Kentucky BOTANY Chairman: Cart. HENRICKSON, University of Kentucky Secretary: ARLAND Horcuxiss, University of Louisville CHEMISTRY Chairman: Cari. Hussune, Murray State College Secretary: AnTHUR W. Fort, University of Kentucky GEOLOGY Chairman: JaMrEs E. Conxr1n, University of Louisville Secretary: Marion STALLARD, Louisville PHYSICS Chairman: Bruce B. VANCE, Louisville Public Schools Secretary: RicHarD Hanau, University of Kentucky PSYCHOLOGY Chairman: Ray H. Bixier, University of Louisville Secretary: Paut McNEEty, Asbury College ZOOLOGY Chairman: RoBERT KUEHNE, University of Kentucky Secretary: Dwicur Linpsay, Georgetown College BOARD OF DIRECTORS a. YAUGANGASTER) 0; ii cscocsscetssectencaes .-to 1961 HAZEL NOLLAU .........cccccccscenscnss ....to 1962 eG RIE yy ke Nd telat aalvadenes ave to 1961 WHET TANS. (CrAS © ii. 00L ea eee to 1963 WILLIAM B. OWSLEY ......cccscesecsecoccees to 1962 CART GANGS (ule. iestukee cose aca uue aay to 1964 Gu Bu FIAMANN) 2552 A eociesese eeee teens to 1962 NOU CPN SAS 0 9a CR Ser pS A EN to 1964 EDITORIAL STAFF Editor: RoGER W. Barsour, University of Kentucky, Lexington, Ky. Associate Editors: t (Bacteriology and Medical Technology) SrrH Gi_krrson, Berea College, Berea. (Botany) Mary E. WuHarron, Georgetown College. (Chemistry ) Warp SuMptTER, Western State College, Bowling Green. (Geology) Barspara M. Conxtn, Louisville : (Zoology) Joun M. Carpenter, University of Kentucky, Lexington aw, Membership in the Kentucky Academy of Science is open to interested persons upon nomi- nation, payments of dues, and election. Application forms for membership may be obtained from the Secretary. The TRANSACTIONS are sent free to all members in good standing. Subscription rates for non-members are: domestic, $2.00 per volume; foreign, $2.50 per * volume. z The TRANSACTIONS are issued semi-annually. Four numbers comprise a volume. ‘A Correspondence concerning memberships or subscriptions should be addressed to the oi | Secretary. Exchanges and correspondence relating to exchanges should be addressed, The 3 Librarian, University of Louisville, who is the exchange agent: for the Academy. Manuscripts and other material tor publication should be addressed to the Editor. SOIL TEMPERATURE MEASUREMENTS AT LEXINGTON, KENTUCKY FROM 1952 TO 1956 E. B. PENROD, J. M. ELLIOTT, and W. K. BROWN Department of Mechanical Engineering, University of Kentucky, Lexington, Ky. Soil temperature variation with time and depth is of interest to the design and maintenance engineer and agronomists. The annual variation of ground temperature must be taken into consideration by engineers in designing buildings, dams, highways, airport runways, water mains, pipelines, underground high voltage cables, and ground coils for earth heat pump installations. The extent of interest in soil science in recent years can be obtained readily by referring to the publications listed in the Bibliography. Research In Geophysics Research in geophysics was started at the University of Kentucky in 1949 when an earth heat pump system was employed to determine the thermal diffusivity of a clayey soil immediately west of Anderson Hall [4, 5].1 The soil is covered with sod and has a general Casa- grande classification of lean clay with low plasticity characteristics. The experimentally determined values of the thermal diffusivity and density of the soil are 0.019 ft? hr and 120 lb. ft-* respectively, and the calculated values of the specific heat and thermal conductivity of the soil are 3.37 B Ib-1 F—1 and 0.84 B hr~ ft-1 F—! respectively. The specific heat of a sample of dried clay, taken from the earth near the ground coil of the heat pump system, was determined by the use of a special calorimeter [13]; the specific heat and density of the dried clay were found to be 0.196 B Ib-! F~- and 95 lb ft-? respectively. From these values, the specific heat of the wet clay was calculated to pe 0.37 Blb-* F-1, Thermal properties of a sample of fine clay were determined in a single transient heat flow laboratory experiment [16]. The clay was taken at a depth of about six feet, east of Anderson Hall during an excavation for a new building, and was dried in an oven at 230° F. The physical properties, of a portion of the dried clay that passed through a No. 10 sieve, are: thermal diffusivity 0.0075 ft? hr-! thermal conductivity O23 Bears tt 1 The numbers in brackets refer to research papers and reports listed in the Bibliography. = PSV) ev » ; Perrine yw JAR 2 i " y Pee heaves t 50 E. B. Penrod, J. M. Elliot, and W. K. Brown specific heat 0.204 B Ib- F-1 density (dry) 80.5 Ib ft-%. Geophysical studies at the University of Kentucky, also, include soil temperature measurements and the determination of solar energy received on a horizontal surface [19, 24]. The empirical equations, developed for calculating soil temperatures at any time and depth, are based on soil temperatures taken at depths of 0, 2, 4,7 5, 6, 8 and 10 ft. west of Anderson Hall. The thermocouple used to determine the surface temperature was buried beneath the sod at a depth of about one-half of an inch. Soil temperatures have been recorded at two- hour intervals since 1952. Empirical Equations The surface temperature of the earth undergoes an annual tem- perature change that is nearly simple harmonic. It is often desirable to have an equation that can be used in calculating soil temperatures at any depth for a particular time. To derive an equation, it is as- sumed that the soil at a given locality is uniform, its surface is flat; and that heat flows in a direction perpendicular to the surface [19]. It is then only necessary to solve the Fourier heat equation C) att Ot Ox= subject to the boundary condition t=t, sin Sv > (2) where t — temperature, t,; — temperature amplitude, 7 = time, P — period, a — thermal diffusivity, and x = distance from the sur- face. A particular solution of Eq. (1) is given by the equation ty=tm + to rate Tr /ex P sin a (T—-Te) —xX =P | ; where t, — temperature at any distance x from the surface, tn = mean surface temperature and/or mean soil temperature, 7, = the time in- terval between the middle of a designated month and the date that the surface temperature is equal to the mean temperature [19]. The following equations were developed, from recorded tempera- 2 The thermocouple used to obtain temperatures at a depth of 4 ft. ceased to function in November, 1953. Soil Temperature Measurements at Lexington, Kentucky 51 ture data, for calculating soil temperature, in °F, for any chosen time and depth for the five year period under consideration: SBe (T=58.08 +20.54 e 71°°* sin(SO°T-10.65°-6.25°x), 1953 T=58.28 + 18.83 e099 sin(30°T-26,.23°-5.21°x), 1954 t=59.01+ 23.50 e°°?"* sin(Z0°T -10.38°-7.28°x), 1955 t=56.86+26.25e°°'™ sin(Z0°t-16.71°-7.73°x), "1956 t=56.48+21.88e~°!2™ sin(30°T-12.78°-6.92° x), and the 1952-1956 norm t=S57.75 421.356 0 sin(30°T-15.72°-6.4.9°x), where 7 = 0, 1, 2, . . ., on April 15, May 15, June 15, . . ., and x is the numerical value of the depth of soil. Example.- As an application of the above equation for the five year norm, the soil temperature, say, at a depth of 13 ft. is desired on September 30, 1961. For this depth and time, x = 18 and 7 = 5.5. Substituting 13 and 5.5 for x and 7; in the equation, the soil tempera- ture is found to be 62.2° F. This value, of course, is for the type of soil in question, and will be approximately the same for a similar clayey soils, providing the latitude and climatic conditions do not vary greatly from those at Lexington. Results Using methods similar to those given in Reference 19, values of the mean soil temperature, surface soil temperature amplitude, and average thermal diffusivity were determined and are recorded in Table 1. Calculations were made also, to determine the variation of ‘ay E. B. Penrod, J. M. Elliot, and W. K. Brown Table 1— Summary of Data for the Five Year Period at Lexington, Kentucky. Total precipitation, inches/year f - 42. Average solar energy, Btu/ft*/doy | 1534.58 | 1584.66 | 1617.84 | 1595.69) (567.51 | 1580.05 | Average thermal diffusivity, ft*/hr | 0.0301 | 0.0433 | 0.0222 | 0.0197 | 0.0245 “Dota supplied by the U.S. Weather Bureau, Blue Grass Airport, Lexington, Kentucky. soil temperatures with time and depth. These results are presented graphically in Figures 1 to 9. By the method of trial and error, the following equation was de- veloped, using observed air temperatures taken and recorded at the U.S. Weather Station at the Biue Grass Airport, Lexington, Kentucky, for the five year period, 1952-1956: tair= 56.55 +2183 sin(Z30°T-2°52'), °F, where 7 = 0, 1, 2, s.., on April 15, May 15, June 15,.... Calculated and observed air temperatures are listed in Table 2. The agreement of the air temperatures calculated by the use of the above equation and the observed values is very good. In Table 3 are listed calculated and observed soil temperatures. The agreement between the calculated and observed soil temperatures are excellent, except near the earth’s surface which is subjected to sud- den changes in weather. In developing the empirical equation the assumption, that the surface soil temperature varied in a sinusoidal manner, is approximately true. The area between the arch of the sine-surface temperature curve (Fig. 1) and the mean temperature axis from May to November is, of course, equal to that for the follow- ing six months. Of the total annual solar energy received by the earth's surface at lexington, 66 per cent is received from May to No- vember and only 34 per cent during the following six months. As a result of this, and probably other conditions, the area between the mean temperature axis and the curve (not shown in Fig. 1), for the observed surface soil temperatures for tne period, November to May, is less than the area corresponding to the period, May to November. Soil Temperature Measurements at Lexington, Kentucky 53 Table 2.— Observed and Calculated Air Temperatures for the Five Year Norm, 1952-1956, at Lexington, Kentucky Date Time | Observed* | Calculated | Difference be- air temp.°F | air temp.°F. | tween obs. and cal. temp°F *Data taken at the Blue Grass Airport, Leneatun, Kentucky by the U.S. Weather Bureau. Table 3.— Calculated and Observed Soil Temperatures for Lexington, Kentucky for 1952* Temperatures in degrees F.. at depths of Oft Zift S tt 6 ft 10 ft Calc Obs | Calc Obs Calc Obs Calc Obs Calc ‘Obs 3789) 41.41 | 42.90) 44.56 5 : ! ; 53.87 |52.44 38.70/40.16 F : : fs Q 4 50.70] 51.04 44.70} 49.79 i : : ; A d 49.50| 49.86 50.60/50.39 53.71 | 52.84 57.98] 57.97 62.29] 62.26 65.46] 65.43 66.66) 67.04 65.56] 66.16 62.45] 62.46 58.18 | 58.07 *Similor tables were made for the other four years, but are not included in this report. 54 E. B. Penrod, J. M. Elliot, and W. K. Brown The temperature curves, shown in Figures 7-9, for May and No- vember cross the mean temperature axis, 57.75° F, at 2 ft. and 30 ft. Therefore, the half wave length and wave-length are 28 ft. and 56 ft. respectively. Similarly, the temperature curves for August and Feb- ruary cross the mean temperature axis at depths of 16 ft. and 44 ft. respectively, showing that the half wave-length and wave-length for these months are also 28 ft. and 56 ft. respectively. Conclusions and Remarks Soil temperatures have been measured and recorded, at two-hour intervals, from 1952 to 1956 inclusive. From the measured or observed data empirical equations were developed for calculating soil tempera- tures for a specified time and depth for each year and also for a five year norm. The variation of soil temperatures with the time are presented graphically for different depths. From these graphs it can be seen readily the temperature amplitudes decrease with depth, and that maxima and minima of temperature shift from left to right as the soil depth increases. The calculated value of the thermal diffusivity of the soil, for all depths, is 0.0279 ft? hr-1 for the five year period. From the graphs showing soil temperature versus depth, the wave- length of the temperature waves was found to be 56 ft. The tempera- ture graphs for each month of the year converge, in a sinusoidal man- ner, to the mean soil temperature of 57.75° F at a depth of about 100 ft. : An empirical equation was developed for calculating the mean air temperature at Lexington for any specified time. The mean air tem- perature and mean surface soil temperatures are 56.55° F and 57.75° F for the five year norm. The graphs in Fig. 6 show clearly the manner in which the air temperature and the surface soil temperature vary with the quantity of solar energy that is received at the earths surface, ~ Soil Temperature Measurements at Lexington, Kentucky 55 TEMPERATURE —°F > 6, ] ro) =SYNLVYSdNSL XIV NVAW GSAYNSSEO SHLNOW-SWIL (<= a a eal | ata an CG PoC NY | PRECIPITATION—INCHES Fig. 1. Variation of soil temperature versus time at different depths. Soil temperatures were calculated by the use of the empirical equation deyeloped for 1952. "4 Bad ‘ (xG2°9-S9'01-2,0E)uIS, 2 >S'0Z+80'8S =} SJ CLs al Zi) in ee Sl ei aN SNC SHLNOW-3WIL DAN he ee Ee a Lae "4 EJS [Eli as Se SN Gs mS NS uu cs (Oa ee 5) 5) ~ | IS (eo) nN > (ep) PRECIPITATION—INCHES Fig. 2. Variation of soil temperature versus time at different depths. Soil temperatures were calculated by the use of the empirical equation developed for 1953. = > a 4620 ‘(XJ2'S— €2°9ZS- 2 OL)UIS yeog9:9-928'SI+82'9S="} =! 56 E. B. Penrod, J. M. Elliot, and W. K. Brown a UBS sei Ne = le, Bo Din m cj ed (os zs lm 5m aS 3 > =v As mo cm 1 D> = m2 ro) | =e 7) = | eg re y. ct a U] on oO = + Nw oe n [o) °, © nm ~_ = = S Oi o, 3 ue o O @ ° I N in) @ ° = iS) a a n PRECIPITATION. = INCHES Fig. 3.— Variation of soil temperature versus time at different depths. Soil temperatures were calculated by the use of the empirical equation developed for 1954. JT EMPERATURE— © SRaSESRHECSaGnE Sa See SS6l FANS ABE ey, (acecnvane7 ZK "4 $68Q ‘(K,€2 LIZ 91-2 OLMIS yo¢j9-2S2'92498'9S= "3 aNd SECC = PRECIPITATION — INCHES Fig. 4. Variation of soil temperature versus time at different depths. Soil temperatures were calculated by the use of the empirical equation developed for 1955. ene Tung Tr b O oO Coos Pe) See A Lae | | e Din a LiMn eee ee m2 a m2 = < Aras 35 > mi o @) 25 Smee ys al | || a= =m em mo DP u = melee mz Slr =" (ial Sees el --> ae) eo | Lee) : | St NA [oS oO S fo) PRECIPITATION— INCHES Fig. 5. Variation of soil temperature versus time at different depths. Soil temperatures were calculated by the use of the empirical equation developed for 1956. PRECIPITATION- INCHES” Ves outs bs) @ oO — N Oo > ui 3 & : Po 3 =) SSS SSS SS = =e es a 4 5 aa Se m x alae 2 ao) = saan 7 “= a ‘ “il oe 2 nN ol SOLAR ENERGY— B-FT™®-DAY'x1I0 Fig. 6. Variation of soil temperature and mean air temperature versus time at different depths. Soil temperatures were calculated from the empirical equations developed for the five year norm. Similarly, mean air temperatures were calculated from the empirical equation developed for the five year norm. The mean soil temperature and mean air tem- perature for the norm are 57.75° F and 56.55° F. 58 E. B. Penrod, J. M. Elliot, and W. K. Brown ; Baie | OY AG: 8 AN RE + SUBERATURES id Fig. 7. Variation of soil temperature versus depth for the five year norm for 1952-1956. Rrange, from the surface to a depth of 22 ft. 72, ‘76 20 BY DEPTH—FEET 40 560 aa 576 58.0 584 °&588 : 59.6 “TEMPERATURE — °F Fig. 8. Variation of soil temperature versus depth for the five year norm for 1952-1956. Range, from 20 ft. to a depth of 46 ft. DEPTH—FEET Fig. Soil Temperature Measurements at Lexington, Kentucky 59 44 48} oi in) On (o2) to) (2) 64 5760 3768 5776 57.84 S792 TEMPERATURE °F 9. Variation of soil temperature versus depth for the five year norm for 1952-1956. Range, from 44 ft. to a depth of 68 ft. to ivy) = Bibliography The Residential Heat Pump In New England by C. H. Coogan, Jr. Connecti- cut Light and Power Bulletin, August, 1948,’ Waterbury, Conn. Ground Temperatures as Affected by Weather Conditions by A. B. Algren. Heating, Piping and Air Conditioning, Vol. 21, No. 6, June, 1949, pp. 111-116. Thermal Properties of Soils by M. S. Kersten. University of Minnesota Engineering Experiment Station Bulletin No. 28, June, 1949, pp. 1-225. Earth Heat Pump Research—Part I by E. B. Penrod, O. W. Gard, C. D. Jones, H. E. Collier and R. E. Patey. University of Kentucky Engineering Experiment Station Bulletin, Vol. 4, No. 14, December, 1949, pp. 1-64. Measurement of the Thermal Diffusivity of a Soil by the Use of a Heat Pump by E. B. Penrod. Journal of Applied Physics, Vol. 21, No. 5, May, 1950, pp. 425-427. Thermal Conductivity of Soils for Design of Heat Pump Installations by G. S Smith and Thomas Yamauchi. Heating, Piping and Air Conditioning, Vol. 22, No. 7, July, 1950, pp. 129-135. Factors Useful in Ground Grid Design for Heat Pumps by G. S. Smith. Heat- ing, Piping and Air Conditioning, Vol. 23, No. 1, January, 1951, pp. 169-176. Climatology as an Aid in Heat Pump Design by G. S. Smith. Heating, Pip- ing, and Air Conditioning, Vol. 23, No. 6, June, 1951, pp. 101-107. Soil Temperature, Moisture Content, and Thermal Content, and Thermal Properties by C. L. Carter. University of Tennessee Engineering Experiment Station Bulletin No. 15, June, 1951, pp. 1-85. 10. it; E. B. Penrod, J. M. Elliot, and W. K. Brown Soil Temperatures In Water Works Practice by R. F. Legget and C. B. Craw- ford. Journal of American Water Works Association, Vol. 44, No. 10, Oc- tober, 1952, pp. 923-939. Frost Action in Soils—A Symposium. Highway Research Board Special Re- port No. 2, National Academy of Sciences, National Research Council Publi- cation 213, 1952, Washington, D. C. Frost Action in Roads and Airfields, A Review of the Literature, 1765 to 1951. Highway Research Board Special Report No. 1, National Academy of Science, National Research Council Publication 211, 1952, Washington, D. C. Specific Heat Measurement by E. B. Penrod and F. M. Wells. Transactions of the Kentucky Academy of Science, Vol. 14, No. 1, February, 1953, pp. 22-32. Sizing Earth Heat Pumps by E. B. Penrod. Refrigerating Engineering, Vol. 62, No. 4, April, 1954, pp. 57-61, 108, 112. Frost Penetration Studies in Canada as an Aid to Construction by C. B. Craw- ford. Roads and Engineering Construction, Vol. 93, No. 2, February, 1955. An Experimental Determination of the Thermal Properties of Clay by E. B. Penrod, G. T. Privon and K. V. Prasanna. University of Kentucky Engineer- ing Experiment Station Bulletin, Vol. 10, No. 39, March, 1956, pp. 1-76. Ground Temperature Investigation In Canada by C. B. Crawford and R. F. Legget. Engineering Journal, Vol. 40, No. 3, March, 1957. Geotechnical Properties of Leda Clay In the Ottawa Area by W. J. Eden and C. B. Crawford. Proceedings of the Fourth International Conference on Soil Mechanics and Foundation Engineering, London, 1957. See also, Re- search Paper No. 37 of the Division of Building Research, Ottowa, Canada. A Method To Describe Soil Temperature Variation by E. B. Penrod, W. W. Walton and D. V. Terrell. Proceedings of A.S.C.E., Vol. 84, Part I, No. SM-1, February, 1958, pp. 1537-1 to 1537-21. Evaluation of Vehicle Sinkage in Off the Road Locomotion by M. G. Bekker. Report No. 35, June, 1958, pp. 1-49. Ordnance Corps, Land Locomotion Research Branch, Research & Development Division, OTAC, Detroit Arsenal, 28251 Van Dyke Avenue, Center Line, Mich. Soil Temperatures by B. J. Fluker. Soil Science, Vol. 86, No. 1, July, 1958, pp. 35-46. Preliminary Study on Jumping and Running Types of Locomotion by R. K. Bernhard. Report No. 438, July, 1958, pp. 1-46, Ordnance Corps, Land Locomotion Research Branch, Research & Development Division, OTAC. . A Soil Value System for Land Locomotion Mechanics by W. L. Harrison et al. Research Report No. 5, December, 1958, pp. 1-94. Army Ordnance Tank-Automotive Command, Center Line, Mich. Solar Energy Measurements at Lexington, Kentucky For 1951 by E. B. Pen- rod, W. W. Walton and H. O. Knight. Transactions of the Kentucky Acad- emy of Science, Vol. 20 (1-2), 1959, pp. 4-10. Solar Energy at Lexington, Kentucky for 1952 by E. B. Penrod and Cho-Yen Ho. Accepted for publication, Transactions of the Kentucky Academy of Science. Soil Temperature Variation at Lexington, Kentucky From 1952-1956 by E. B. Penrod, J. M. Elliott and W. K. Brown. Available for publication. Accepted for publication 15 January 1960. CYCLIC ENTRY-EXIT FOR SUBROUTINES J. C. EAVES and T. J. PIGNANI Department of Mathematics and Astronomy University of Kentucky, Lexington, Ky. Introduction The purpose of this paper is to call attention to the Cyclic Entry- Exit Method for subroutines used with high speed computers. We shall point out some of the advantages in using available subroutines. We shall outline the general principle followed in making a subroutine an integral part of one’s own particular program. The merits of the Cyclic Entry-Exit Method will be discussed. For contrast with the usual treatment of this problem, the reader may wish to refer to the papers of Livesley (1957), McCracken (1957), and Wrubel (1959). Whenever it is necessary to refer or make use of a computer instruction in this work we shall confine ourselves to the basic machine language, that is, the absolute codes, used for programming the IBM type 650 Magnetic Drum Data-Processing Machine. Subroutines are actually prepackaged special purpose programs. Subroutines are becoming more in demand since the number of computer patrons has increased almost beyond reasonable expectation. Users represent virtually every field of study found on a university campus in addition to a variety of business and industrial activities. Inexperienced programmers, as well as the experts, have brought about a crutial demand for effective, efficient subroutines. Justification for the use of existing subroutines seems to be well founded in the following advantages: 1. Time saving. By avoiding duplication of program writing time a better utilization of programmer efforts is achieved. 2. Better program. A subroutine which has been carefully worked out and used is likely to be more nearly optimized than the usual program given its first run This saves computer time. 3. Debugged. Subroutines available for use have been run and checked. The programmer may save considerable debugging time. 4. Acceptability. Most subroutines are supplied with established computational limitations and the user knows, through the error analysis studies of this particular subroutine, the acceptability and goodness of computed results. 62 J. C. Eaves and T. J. Pignani Subroutine Entry-Exit Principle A subroutine may be a correction routine needed for the correction of a program already underway or it may be a routine which causes the computer to perform a supplementary computation. In using a subroutine one attempts to make the subroutine an integral part of the main program. Thus the computer uses instructions from the main program until such time as those instructions contained in the sub- routine are needed. The main program instruction just prior to entry into the subroutine must contain three items of information; namely, the operation to be carried out just prior to the use of the subroutine, the cell location affected by this operation, and the location of the first instruction of the subroutine. The final instruction of the sub- routine usually directs the computer to leave the subroutine. The instruction address of this last subroutine instruction usually gives the cell location of the desired main program instruction to be executed immediately following completion of the subroutine. The Cyclic Entry-Exit Method For purposes of illustration we shall assume that our main program is to be stored in cells 0200 through 0499. We shall assume that the subroutine to be used is punched to be stored in cells 1800 through 1847. Furthermore, we shal] assume that this subroutine is to replace instructions located in cells 0400 through 0478. The Cyclic Entry-Exit Method then requires the following modifications of instructions. 1. Replace the main program instruction xx yyyy 0400 of cell 0399 with the instruction. xx yyyy 1800. This directs the computer to cell 1800 for its next instruction after the completion of operation xx. 2. The first instruction of the subroutine, located in cell 1800, is 69 0479 1801 which causes the computer to load the distributor with the instruction located in cell 0479. It should be noted that this is the first main program instruction to be used following the completion of the subroutine. 3. The second instruction of the subroutine is 24 1800 1802. This causes the computer to place the instruction, formerly in cell 0479, which is to be used immediately following completion of the sub- routine, in the first cell used for the subroutine. The computer is then directed to ce!l 1802 where it obtains the first computational instruction of the subioutine. The computer procedes through the subroutine computations in the prescribed manner until it encounters the last instruction of the subroutine (which may not be in cell 1847 due to optimization ). Cyclic Entry-Exit for Subroutines 63 4. The last instruction of the subroutine, written at the time the subroutine is prepared, directs the computer to get its next instruction from cell 1800. Thus the computer actually returns to the beginning of the subroutine only to find that the first instruction of the subroutine has been replaced by the de-ired instruction of the main program. This exit instruction is of the form xx yyyy 1800. It should be noted that the insertion of a set of instructions is achieved by exactly the same procedure used for the replacement of a set of instructions. One merely has the first instruction of the subroutine direct the computer to the desired instruction to be per- formed immediately following the execution of the subroutine. Using the above illustration the first instruction of the subroutine would be xx yyyy 0400 in case we wish to insert the subroutine between the main program instructions carried in cells 0399 and 0400. Generalized Use of Cyclic-Entry-Exit Method Multiple use of the Cyclic-Entry-Exit Method for the insertion of several subroutines and the replacement of parts of the original pro- gram is ‘'emonstrated by the brief program below. In this skeleton program ;resented, we insert the subroutine in three places and furthermore the subroutine replaces those instructions in cells 0400 through 0478, of the program used in the former section. Main Program Revisions Cell Instruction Replacement Instruction 0200 0257 xx yyyy 0258 xx yyyy 0400 0258 0290 xx yyyy 0291 xx yyyy 0401 1 0350 xx yyyy 0351 xx yyyy 0402 035% 0399 xx yyyy 0400 xx yyyy 0403 0400 xx yyyy 0401 69 0258 1801 0401 xx yyyy 0402 69 0291 1801 0402 xx yyyy 0403 69 0351 1801 0403 xx yyyy 0404 69 0479 1801 0404 Delete Replaced cath hee by 0478 Delete Subroutine 0479 0499 64 J.C. Eaves and T. J. Pignani Subroutine Cell Instruction 1800 00 0000 0000, exit cell to be used as needed 1801 24 1800 1802 1802 First instruction of subroutine 1847 xx yyyy 1800 A study of the above revised program shows that each time the program refers to the subroutine the operation to follow subroutine activity is stored in cell 1800. Also, part of that bank of cells deleted from the program is utilized for subroutine entry. The insertion of several different subroutines is made by a very similar adjustment in the program. Advantages of the Cyclic Entry-Exit Method 1. The replacement and insertion of instructions are carried out in the same way. 2. The programmer need not concern himself with the where- abouts of the last instruction of the subroutine since the exit is made through the entry cell. 3. Cyclic Entry-Exit Method requires only one alteration of the subroutine deck whereas other subroutine entry-exit techniques require the alteration of two or more of the subroutine instructions. 4, This method requires the use of only two extra operations and two extra cells of storage in the subroutine whereas many modified entry-exit programs make additional demands on storage and opera- tion time. Especially is this true with those methods requiring accum- ulative additive features or instruction modification. Literature Cited Livesley, R. K. 1957. DIGITAL COMPUTERS. Cambridge University Press: 27, 40. McCracken, D. D. 1957. DIGITAL COMPUTER PROGRAMMING. John Wiley and Sons: 111, 128, 230. Wrubel, Marshal H. 1959. PROGRAMMING FOR DIGITAL COMPUTERS. McGraw-Hill Book Company: 86, 151, 212. Accepted for publication 23 March 1960, THE SALTING EFFECT OF SODIUM CHLORIDE ON THE EXTRACTION OF ERBIUM ACETYLACETONATE AND 8-QUINOLINOL Y. G. ISHIDA, J. F. STEINBACH and W. F. WAGNER Department of Chemistry, University of Kentucky, Lexington, Kentucky The use of salting agents has proved to be very effective in the solvent extraction of some metal salts, such as, the nitrates by tributylphosphate (Hesford and McKay, 1958), uranyl nitrate (Fur- man, Mundy, and Morrison), thorium nitrate (Bock and Bock, 1950), and ferric chloride (Morrison, 1950) by ether. Morrison and Freiser (1957) have summarized the effect of salting agents on the extraction of inorganic salt systems. Diamond (1957) has derived general equations for the solvent extraction of inorganic compounds and studied the effects of salting agents on the extraction of indium (IIT) chloride from aqueous hydrochloric acid by various organic solvents (Diamond, 1959). Very little work on the effect of salting agents on the extraction of chelates has been reported. Recent studies (Brown, Steinbach, and Wagner, 1960), on the solvent extraction of rare earth acetylacetonates by acetylacetone led to this study of the effect of salting agents on the system. In addition, the extraction of 8-quinolinol (oxine) in a water- chloroform system was studied to compare the effect of salting agents on the neutral oxine, oxinate ion, and oxinium ion at different acidities. Lacroix (1947) has studied the properties and extraction of oxine by chloroform from aqueous solutions as a function of pH from 0 to 14. From potentiometric titration curves he showed that the oxinium ion (protonated oxine) exists in strongly acidic solutions, the neutral oxine is formed almost exclusively in the pH range of 6 to 9 and the oxinate ion is formed in more basic solutions. Extraction data showed an increase in extraction from 0.599, at a pH of 0 to quantitative extraction in the range of 5 to 6, then a decrease to 5°% at a pH of 14. Apparatus and Materials A Beckman Model DK-2 recording spectrophotometer was used for all absorption spectra. A Beckman Model G pH meter was used for all pH measurements. Eastman 8-quinolinol was recrystallized twice from alcohol-water solution to give a product that melted at 74-75°. Acetylacetone obtained commercially was washed once with 10° ammonia solution, twice with water, and distilled; the fraction boiling between 135-138° was used. Erbium acetylacetonate was pre- 66 Y. G. Ishida, J. F. Steinbach, and W. K. Wagner pared by the method of Stites, McCarty, and Quill (1948) and was recrystallized twice from 95°% ethanol. All other chemicals met A.C.S. specifications for reagent grade. Experimental Solubility of Acetylacetone in aqueous solutions of potassium chlo- ride and sodium chloride. Solutions containing varying amounts of the salts were prepared by dissolving the salt in water saturated with acetylacetone in 25-ml. volumetric flasks. The solutions were shaken and permitted to stand overnight at 25° and the excess acetylacetone which separated was removed. The concentration of acetylacetone in the solution was determined spectrophotometrically as iron acetylace- tonate (Steinbach, 1953). One ml. of the solution was added to a 25-ml. flask containing 5 ml. of a solution of ferric sulfate (20 mg./ml.) and 10 ml. of a sulfuric acid-sodium sulfate buffer (pH = 0.5) and diluted to volume. The absorbance minimum of iron acetylacetonate at 700 my was used to plot a working curve for the concentration of acetylacetone. Alternatively, a 1:50 dilution of the sample was prepared and the absorbance of one to five ml. of the diluted sample was used to determine the concentration of the acetylacetone from a working curve prepared at the maximum absorbance of iron acetylace- tone at 482 mp. Distribution of erbium acetylacetonate between acetylacetone and aqueous solutions of sodium chloride. Ten ml. of a solution of erbium acetylacetonate in water-saturated acetylacetone was shaken with 10 ml. of the acetylacetone-saturated water containing the salting agent. The phases were allowed to come to equilibrium overnight in a 25° bath. The erbium present in the organic phase was determined spectrophotometrically by measuring the absorbance of erbium acetyl- acetone at 378 mp. The erbium present in the aqueous phase was determined spectrophotometrically by the arsenazo method of Fritz, Richard, and Lane (1958). Volumetrically the erbium in the aqueous phase was determined by adding an excess of standard 10°? M EDTA and back titrating with standard 10° M zinc solution using PAN indicator. Distribution of oxine between chloroform and water. Ten ml. of 0.1 M oxine in chloroform was shaken for an hour with 10 ml. of the solution of sodium chloride in water and placed in a bath at 25° for 24 hours to reach equilibrium. The concentration of oxine in the aqueous phase was determined by acidifying the solution to a pH of 1 and measuring the absorbance at 251 mp. The concentration of oxine in the organic phase was obtained by difference. The Salting Effect of Sodium Chloride 67 Results and Discussion Solubility of Acetylacetone. Data for the solubility of acetylace- tone in aqueous solutions of sodium chloride and potassium chloride at 25° are shown in Table I and Figure 1. The solubility of acetyl- ais as leit ale 9/ mi. alo a08 0.06 Acetylacetone O04 02 [oxe,e) 10 2.0 3.0 4.0 5.0 Salt concentration MOU/i +6, Fig. 1. Solubility of Acetylacetone in solutions of NaCl and KCI at 25°. Table I.— Solubility of Acetylacttone in Solutions of NaCl and KCI at 25 NaCl KCl NaCl CsHsO:z KCl CsHsOz Mol/1. pH g/ml mol/1. pH g/ml 0.000 4.68 0.173 0.000 4.68 0.173 0.034 4.67 0.169 0.032 4.65 0.169 0.206 4.60 0.159 0.215 4.63 0.160 0.684 4.49 0.138 0.698 4.61 0.142 2.05 4,34 0.094 1.81 4.59 0.110 3.64 4.25 0.065 3.388 4.57 0.075 4.87 4,23 0.052 68 Y. G. Ishida, J. F. Steinbach, and W. K. Wagner acetone in solutions of sodium chloride is slightly lower than in solu- tions of potassium chloride, probably due to the lower activity of the water containing the more highly hydrated sodium ion. In the spectrophotometric determination of acetylacetone it was found that the absorbance minimum for iron acetylacetone at 700 mp shifts to higher wave lengths with an increase in concentration and Beer's Law is not obeyed. Consequently, the absorbance maximum for iron acetylacetonate at 482 mp» where Beer’s Law is followed was used for most analyses. Distribution of erbium acetylacetonate. The data for the extraction » of erbium acetylacetonate from aqueous solutions of 0.1 M and 1.0 M sodium chloride as a function of pH are shown in Table II and Figure 2. The extraction curve in 0.1 M NaCl was practically identical to that without any salt present, while in 1.0 NaCl the curve was shifted OQ----0 EXTRACTION CURVE NO NACL w%---X EXTRACTION CURVE O1M NACL @—e EXTRACTION CURVE 10M NACL Fig. 2. Extraction of Erbium Acetylacetonate as a function of pH. The Salting Effect of Sodium Chloride 69 Table Il._— Extraction of 5.48 x 10-8M Eribium Acetylacetonate as a Function of pH from 0.IM and IM NaCl at 25° Water 0.1IM NaCl 1.0M NaCl pH GE pH TE pH Jo 4.62 22.6 4.61 22.6 4.61 24.3 4.75 27.9 4.77 28.3 4.69 28.5 4.85 36.4 4.85 34.7 4.79 34.3 4,97 39.8 4.96 38.9 4.89 46.0 5.14 44.9 oelul 45.6 4,99 51.4 5.30 52.0 525 2a 5.09 57.0 5.40 Pptd. to lower pH values by about 0.10 units at a pH of 4.61 and by about 0.25 units at a pH of 5.30. A decrease in solubility of acetylacetone in the aqueous phase with increasing concentration of sodium chloride should cause the extraction curves to shift to higher pH values. The shift may be calculated from the equation Kp. Kk [HR]? D = ———_ PEt? where D is the distribution coefficient, K; is the formation constant of the chelate, p. is the partition coefficient of the chelate, K; is the dissociation constant of the acetylacetone, and [HR] is the concentra- tion of acetylacetone in the aqueous phase. In 0.1 NaCl, with other factors constant, the change in concentration of acetylacetone would raise the pH of extraction approximately 0.02 units between a pH of 4.61 and 5.30; in 1.0 M NaCl the shift would be +0.12 units at a pH of 4.61 and +0.14 units at a pH of 5.30. Since the experimental curve showed little if any shift in 0.1 M NaCl it is apparent that 0.1 M NaCl had very little effect on the distribution coefficient. However, in 1.0 M NaCl the experimental curve was shifted to lower pH values. An increase in ionic strength causes a decrease in the formation constants of chelates and an increase in the dissociation constants of weak acids. The presence of sodium chloride in the water makes that phase much less attractive for the chelate, increasing the partition coefficient in favor of the organic phase. Thus, in 1.0 M NaCl the changes in the partition coefficient and the dissociation constant of acetylacetone cause a shift to lower pH values which more than compensates to the shift to higher values by the change in the formation constant of the chelate and decreased solubility of acetylacetone in the aqueous phase. Because buffered solutions are required in counter-current separa- tions the effect of buffers on the extraction of erbium acetylacetonate was investigated. The addition of a sodium acetate-acetic acid buffer 70 Y. G. Ishida, J. F. Steinbach, and W. K. Wagner lowered the per cent of erbium extracted at a given pH. It has been suggested (Brown, 1959) that this shift of the extraction curve to higher pH values is due to the decrease in reagent concentration in the aqueous phase by increasing the ionic strength of the solution. However, the addition of 0.05 M NaCl to the buffered solution raised the per cent of erbium extracted. The shift of the extraction curves to higher pH values in buffered solutions is probably caused mainly by the association of acetate ion with some of the erbium to reduce its extraction into the organic phase. When potassium acid phthalate- sodium hydroxide buffer was substituted for the acetate buffer at the same pH and ionic strength, the per cent erbium extracted at a pH of 5.1 was 32% as compared to 47% for the acetate buffer, and 50% for an unbuffered solution. The lower extraction in the phthalate buffer probably is caused by the greater extent of complex formation between the phthalate ion and erbium. The addition of 0.05 M NaCl to the phthalate buffered system improved the extraction. Distribution of oxine. Data for the extraction of oxine by chloro- form from acidic, approximately neutral, and basic solutions of sodium chloride are shown in Table III. The addition of sodium chloride had Table IIl._— Extraction of 10-2 M Oxine by Chloroform From Acidic, Neutral, and Basic Solutions of Sodium Chloride pH NaCl, m/1. %E Dis) 0.00 20.0 2.5 0.05 31.3 2.6 0.50 35.0 2.6 1.00 39.0 DNAS) 2.70 61.9 6.5 0.00 99.5 6.7 0.05 99.6 6.6 0.50 99.6 6.8 1.00 99.6 6.9 2.70 99.7 9.8 0.00 62.2 9.8 0.05 60.1 10.2 0.50 61.9 10.6 1.00 62.6 10.8 2.70 65.4 the greatest effect on the extraction of oxine from an acidic solution where the oxinium ion predominates; the per cent extracted increases from 30% to 60% upon the addition of 2.7 M NaCl. In the pH range 6.5 to 6.9 where the oxine exists essentially as the neutral molecule, the addition of sodium chloride has very little effect since the oxine is almost quantitatively extracted in this range. In the basic solutions The Salting Effect of Sodium Chloride 7a the addition of sodium chloride in concentrations of 0.05 to 0.5 caused a small decrease in extraction of the oxine which, however, increased with an increased in concentration of sodium chloride. The distribution of oxine may be calculated from the equation (11): Kp D = ———_——_- [H+] Ky a Ky [H*] where D is the distribution ratio, K, is the partition coefficient of the oxine, K, is the dissociation constant of the oxinium ion, HsOxt = H+ + HOx and Kz is the dissociation constant of oxine HOx — Ht -+- Ox-. In acidic solutions where the H.,Ox* ion predominates, the third term in the denominator is negligible and the distribution is dependent on K,. An increase in ionic strength increases the value of K, causing an increase in D which, with an increase in Kp by the salting effect of the sodium chloride, causes a marked increase in the extraction of oxine. In basic solutions where the oxinate ion pre- dominates, the first term in the denominator is negligible and the distribution is dependent on K». An increase in ionic strength increases the value of K:, causing a decrease in D at lower concentrations of sodium chloride. As the concentration of sodium chloride is increased the salting effect, by increasing Ky, more than compensates for the increased K», and the extraction of oxine increases. Summary The solubility of acetylacetone in solutions of sodium chloride and potassium chloride was determined. Extraction of erbium acetylacetonate by acetylacetone from the aqueous phase was shifted to lower pH values by the addition of sodium chloride. The increases in the partition coefficient of the chelate and dissociation constant of acetylacetone by increasing ionic strength more than compensates for the shift to higher pH values by the decrease in concentration of acetylacetone and decrease in formation constant of the chelate. The addition of sodium chloride improves the extraction of oxine from water by chloroform in acidic solutions, has a small effect in basic solution, and has very little effect in neutral solutions. The influence of ionic strength on the dissociation constants of oxine and on the partition coefficient was used to explain the results. 72 Y. G. Ishida, J. F. Steinbach, and W. K. Wagner Acknowledgement The authors thank the Atomic Energy Commission for generous financial support of this work on Contract No. AT (40-1) 2124. Literature Cited Bock, R. and Bock, E., 1950. Z. anorg. Chem., 263: 146. Brown, W. B., 1959. Ph.D. Dissertation. University of Kentucky, Lexington, Kentucky. Brown, W. B., Steinbach, J. F., and Wagner, W. F., 1960. J. Inorg. Nucl. Chem. Isie WIS), Diamond, R. M., 1957. J. Phys. Chem., 61: 69. Diamond, R. M., 1959. J. Phys. Chem., 63: 659 Fritz, J. S., Richard, M. J., and Lane, W. J., 1958. Anal. Chem., 30: 1776. Furman, N. H., Mundy, R. J., and Morrison, G. H., U. S. Atomic Energy Commission Report: AEC D-2938. Hesford, E. and McKay, H. A. C., 1958. Trans. Faraday Soc., 54: 578. Lacroix, S., 1947. Anal. Chim. Acta, 1: 260. Morrison, G. H., 1950. Anal. Chem., 22: 1388. Morrison, G. H. and Freiser, H., 1957. Solvent Extraction in Analytical Chemistry. New York: John Wiley and Sons, Inc., p. 43. __....... Steinbach, J. F., 1953. Ph.D. Dissertation. University of Pittsburgh, Pittsburgh, Pennsylvania. Stites, J. G., McCarty, C. N., and Quill, L. L., 1948. J. Am. Chem. Soc., 70: 3142. Accepted for publication 27 August 1960. A NEW NORTH AMERICAN LEAFHOPPER PREVIOUSLY CONFUSED WITH TYPHLOCYBA ANDROMACHE McATEE (HOMOPTERA CICADELLIDAE) PAUL J. CHRISTIAN Department of Biology, University of Louisville, Louisville, Kentucky Contribution No. 30 (New Series) from the Department of Biology, University of Louisville. Introduction In 1926 W. L. McAtee revised the North American species of the genus Typhlocyba and described in this revision a new species which he named Typhlocyba andromache, from one male specimen collected in Salem, New York, June 27, 1924. In 1942 J. T. Medler redescribed this species and recorded it from Minnesota specimens. When McAtee described the species his method of preserving the cleared dissection was to fasten it to the same point to which the rest of the insect was glued, with a mixture of glycerin and shellac (McAtee 1926, p. 3). At the time he indicated uncertainty as to how suitable this method of preservation would be. The author found on reexamining McAtee types that this material was difficult to remove. In working on the type of T. andromache it was possible to clean the pygofer sufficiently to draw the posterior margin, but the aedeagus being much smaller and more delicate could not be cleaned suitably for observation. The specimens Medler used were in good condition, and since no other specimens of this species could be found in North American collections the author used one of these in his description of this species in his revision of the genus Typhlocyba (Christian, 1953). The posterior margin of the pygofer of the type was not exactly like that of Medler’s specimens, but the degree of difference was such as might be attributed to individual variability. The aedeagus of Medler’s specimens also differed from the description of the aedeagus of the type which was described as being bifid at the apex, although Medler had described his specimens as being bifid, because the sclerotized portion of the shaft appeared bifid and the transparent portion was overlooked. Since McAtee had overlooked the transparent portion of the aedeagus in other species of Typhlocyba (rubriocellata, niobe) the author assumed that he had done the same in this case and that Medler’s specimens were Typhlocyba andromache. 74 Paul J. Christian In the spring of 1957 the author collected from Carpinus a large series of specimens which appeared to be an undescribed species of Typhlocyba resembling T. andromache in the author's revision but differing in the shape of the aedeagus and posterior dorsal angle of the pygofer. On comparing the pygofer with a drawing of the type of T. andromache and the original description of the species it was con- cluded that these were specimens of T. andromache and what had been described as andromache was an unnamed species. The following descriptions of these two species should help workers in identifying them. I wish to express my appreciation to Dr. G. W. Byers for the loan of specimens from the Snow Entomological Museum of the University of Kansas, and to Dr. F. F. Cook for loan of specimens from the University of Minnesota collection. . To beameri Figs. 1-2.— Male genitalia of Typhlocyba andromache and T. beameri. a. Left side of pygofer. b. Left side of aedeagus. c. Posterior aspect of aedeagus. d. Ventral aspect of right style. A New North American Leafhopper 1) Typhlocyba beameri n. sp. (Figs. 2a,b,c,d.) Typhlocyba andromache, Medler, 1942, Minnesota Agr. Exp. Sta. Tech. Bull., 155:144-5. (in error) Typhlocyba andromache, Christian, 1953, Bull. Univ. Kansas Sci. Bull., 39( 11,9) :1176, pl. 83, fig. 4a,b,c,d. (in error) Resembling Typhlocyba andromache McAtee in shape of pygofer but with dorsal posterior angle acute forming less than a right angle, and with apex of aedeagal shaft acute as in Typhlocyba surcula De- Long and Johnson. Length.—3.0-3.25 mm. Color.—Dorsum pale yellowish-white; fore wings subhyaline to cross-veins, apical cells hyaline, slightly fumose, without distinctive markings. Genitalia.—_Male pygofer: in lateral aspect with posterior margin nearly straight, dorsal angle acute, slightly less than a right angle, ventral angle slightly more than a right angle and forming a small ventrally-directed hook. Aedeagus: with atril processes broad, elongate, exceeding shaft in length, arising from base close to shaft, gradually diverging laterodorsad from base, slightly sinuate, reduced to acute apices on outer fourth; shaft arising from base slightly above bases of processes, laterally broadened, margins foliaceous, broadest at gonopore, nearly transparent and extending beyond gonopore as a thin plate with acute apex. Styles: broadly tapering to near apex, abruptly reduced to sharp apices. Types.—Holotype male and five paratype males, Itasca County, Minnesota, July 26, 1939, J. T. Medler, in the Snow Entomological Museum of the University of Kansas; two paratype males, July 26, 1939, and one paratype male July 28, 1939, Itasca County, Minnesota, J. T. Medler, in the collection of the University of Minnesota; one male paratype, Sturgeon Bay, Wisconsin, July 29, 1951, D. A. Dever, in the collection of the author. Typhlocyba andromache McAtee (Figs. 1a,b,c,d.) Typhlocyba andromache McAtee, 1926, Proc. U. S. Nat. Mus., 68(18): 82., fig. 67. Resembling Typhlocyba beameri in the shape of the posterior margin of the pygofer, but differing in having the dorsal posterior 76 Paul J. Christian angle slightly rounded, forming not less than a right angle, and having apex of aedeagal shaft bifid. Length.—3.5 mm. Color._Head, pronotum and _ scutellum white to pale yellow, tegmina lemon yellow, subhyaline to cross veins, apices hyaline, slightly fumose, without distinctive markings. Genitalia._Male pygofer: in lateral aspect, with posterior margin nearly straight, dorsal angle slightly rounded, forming a right angle, ventral angle slightly more than a right angle and forming a small ventrally-directed hook. Aedeagus: with atrial processes slender, slightly enlarged near apical third, gradually tapering to sharply acute apices, slightly diverging dorso-laterad from base, apices slightly sinuate in lateral aspect, exceeding shaft in length, distinctly separate from shaft from base, shaft of nearly uniform width, gradually tapering to slightly inflated bifid apex, gonopore apical. Styles: slender, grad- ually tapering to needlelike apices. Material studied._Holotype male, Salem, New York, June 27, 1924, E. D. Ball; a large series of male and female specimens, Louisville, Kentucky, May 1957, collected on Carpinus sp. by the author. A female specimen taken in copula with the male used in making the drawings of the male genitalia of this species in this paper, collected May 30, 1957, Louisville, Kentucky, here designated Neallotype, is deposited in the Snow Entomological Museum of the University of Kansas. Literature Cited Christian, P. J., 1953. A Revision of the North American Species of Typhlocyba and its Allies (Homoptera, Cicadellidae). Bull. Univ. Kansas Sci. Bull. 35(IL9):1103-1277, pls. 73-92. McAtee, W. L., 1926. Revision of the American Leaf Hoppers of the Jassid Genus Typhlocyba. Proc. U. S. Nat. Mus., 68(18):1-47, pls. 1-6. Medler, J. T., 1942. The Leafhoppers of Minnesota (Homoptera: Cicadellidae). Minnesota Agr. Exp. Sta. Tech. Bull. 155:1-196, pls. 9. Accepted for publication 6 February 1960. STUDIES OF THE OVERWINTERING POTENTIAL OF CERTAIN DROSOPHILA SPECIES WALLACE D. DAWSON Present address: Dept. Zoology, Ohio State University This research was supported in part by grants from the University of Kentucky Faculty Research Fund and The National Science Foundation. Introduction Many investigators, both in this country and abroad, have found that Drosophila populations tend to exhibit a characteristic seasonal population frequency cycle. Patterson (1943) in collections at the Aldrich farm near Austin, Texas, over a three year period showed that D. melanogaster reached a peak population in September, while D. affinis-algonquin frequencies were high during the cooler months of the year, but extremely low during the mid-summer months. D. putrida populations tended to vary with rainfall, being high during periods of high precipitation. Williams and Miller (1952) in Nebraska and Carpenter and Giordano (1955) in Tennessee demonstrated similar seasonal fluctuations. Mather (1956) showed high summer frequencies for D. melanogaster in Queensland, Australia, as did Takada (1957) in Japan. The various possible causal factors of these seasonal changes have elicited interest from several investigators. Most attention has been given to climatological, nutritional and genetic factors. Patterson (1943), Speiss (1949), Levitan (1954), Jones (1958) and others found that high frequencies of D. melanogaster and putrida can be correlated with temperature and precipitation increases, respectively. Studies by Komatsu (1959) indicate that food selectivity is not a major factor in controlling the size of Drosophila populations. Dubinin and Tiniakov (1946), Dobzhansky (1956), Carson and Stalker (1949) and Levitan (1951) have studied seasonal changes in inversion frequencies of several species of Drosophila, but whether they are of any sig- nificance in the determination of population density has not been determined. The availability of suitable breeding sites, as suggested by Carson and Stalker (1951) and Carson, Knapp and Phaff (1956) may be a limiting factor. The various seasonal studies have suggested that there may be differences in the ability of the various flies to survive the winter, which largely determine the subsequent behavior of the population during the remainder of the year. The high frequency of D. affinis- 78 Wallace D. Dawson algonquin in the spring and late autumn indicates that this fly may be better adapted to cooler temperatures than other native and domestic species. D. affinis-algonquin may survive the winter in greater numbers than other species and, therefore, have a greater initial reproducing population during the first warm days of spring. On the other hand, such species as D. melanogaster, which are rare in the spring but attain a high population in August and September, may not survive the winter in natural habitats, or else may survive in extremely low numbers, and thus may have to reestablish their populations annually from domestic situations, or, alternatively, require several months to attain sufficient numbers of reproducing flies to establish a peak population. In this study two approaches to the overwintering problem were undertaken. First, field collections for adult Drosophila in natural wooded habitats were made during the winter, particularly during brief mild spells. Materials which might contain overwintering eggs, larvae or pupae were also collected in the field. Second, various species were directly tested in the laboratory under controlled condi- tions by exposure to low temperatures for given periods of time. Such investigations should provide information on the comparative toler- ances of these species to cold, which could then be correlated with the seasonal population fluctuations observed during warmer periods of the year. Materials and Methods Collecting Methods. A wooded area seventeen miles southeast of Lexington, Kentucky, near the Kentucky River was selected as a favorable location for the winter collection of flies. The area was designated Dry Branch Field Station. This location is remote from any urban influences, with human habitation in the area being limited to two small farm tenant houses. Collecting was not done in the immediate vicinity of these dwellings. There are no considerable garbage disposals, orchards, gardens or other similar factors which might affect Drosophila populations in the area. Deciduous trees and red cedar, together with a wide variety of undergrowth compose the vegetative cover. For collecting, a modification of the bottle method outlined by Dobzhansky (1936) was utilized. One-half pint milk bottles baited with a slant of banana-malt-agar mixture were inserted in the forks of trees and other suitable locations at a number of separate points within the area. After a week or ten days flies were collected by inserting a gauze covered cotton stopper in the neck of the bottle. The entrapped flies were returned to the laboratory for identification. Studies of the Overwintering of Certain Drosophila Species 79 Collection records included number of flies of each species collected, climatological data and miscellaneous other information, including observations on the condition of the flies. Collection of materials which could possibly contain overwintering eggs, larvae, or pupae consisted of selecting a variety of likely ma- terials, such as decaying material from tree stumps, walnut hulls, fruits, etc., and placing these in a carton with a small amount of culture medium and holding them at optimum temperatures for two or three weeks, then observing whether adult Drosophila were present. Techniques for Low Temperature Studies. Drosphila stocks of the species affinis, robusta, melanogaster, immigrans, hydei, cardini and tripunctata were maintained in culture for use in low temperature studies. All stocks except hydei and cardini were derived originally from wild flies taken in collections at Dry Branch during the summer of 1958. The hydei culture was obtained from domestic flies in the laboratory. The cardini stock was derived from several flies taken from a swamp near Orlando, Florida. Most of the low temperature experi- ments involved affinis, robusta and melanogaster, therefore, only these species were cultured extensively. Flies were all cultured on a medium of corn meal agar fortified with sugar and dried yeast with occasional modifications. For many of the low temperature tests it was desirable to have flies of a known age range in order to standardize the experiments as much as possible and to minimize errors in the results due to natural death. Aged flies were obtained by clearing thriving culture bottles, in which adults were ecluding from the pupae in large numbers, of all adult flies. These bottles were then held separately for thirty-four hours and all adults which had emerged in the interval fell within the desired age range. These flies were lightly etherized and examined prior to placing in testing bottles. Flies which showed structural defects and which were very recently emerged were rejected. Flies were chosen randomly from the remainder keeping sex ratio approxi- mately equal. For the purpose of exposure to low temperature flies were placed into half-pint milk bottles which had been specially prepared. Each bottle had at the bottom a moist square of bibulous paper thoroughly innoculated with a yeast suspension upon which the flies could feed prior to testing. Any excess moisture was removed prior to placing the flies in the bottles. A dry elongated folded strip of bibulous paper was inserted along one side of the bottle. The bottle was positioned with this side down, and the flies while yet under etherization were placed upon this folded strip in the desired numbers. The bottles 80 Wallace D. Dawson were closed with a gauze covered cotton stopper. The flies were then permitted to revive in the bottle and fed for a period of two hours. Prior to exposure a period of acclimatization at +7° C. (1°) was allotted for a period of time, generally overnight. For the low temperature experiments the freezer chest of a standard commercial eight cubic foot refrigerator was used. This equipment had certain limitations, the principal one of which was difficulty in maintaining an exact constant temperature, especially at the lower ranges used, thus the resultant data lists temperatures in ranges, for example “—5° C. range” is equivalent to —5° C. (2°). Following the acclimatization period flies in the testing bottles were placed in the freezer chest for the desired time. During this time occasional random readings were made from a thermometer in the chest to insure that the temperature did not vary from the desired range. After a given period in the freezer the flies were removed and a four hour period at +7° C. was allotted for deacclimatization since rapid warming appeared to reduce survival. Following this the bottles were removed to room temperature and permitted to stand for one hour. The flies were then emptied into a 10 x 16 x 2 inch white enamel pan for checking. Flies were examined and categorized into four rather well defined major groups based upon the degree of damage to the fly: Complete survival: Flies which behaved normally and displayed regular flight when stimulated. The criteria used for defining this category was the ability of the insect to fly from the floor of the pan with a sustained flight. Crippled: Flies which were capable of any degree of walking, but which were unable to maintain a sustained flight sufficient to escape from the pan. Stunned: Flies which were not capable of locomotion, but dis- played movements and twitchings when observed closely with a binocular microscope. Dead: Flies which displayed no movements or other signs of life when observed closely under a binocular scope. Since at least some of the flies in the crippled category as well as those which completely survived exhibited reproductive capability following cold treatment, both of these categories were considered as survival. The stunned flies inevitably died within a few hours without any degree of recovery, and therefore, these together with the dead were considered as non-survivals. | Studies of the Overwintering of Certain Drosophila Species 81 Results Overwinter Collections. In the course of the two year overwinter studies 1193 Drosophila flies representing ten species were collected at the Dry Branch area. Of this number 308, representing six species, were taken during the critical months, December through March. The remainder of the flies were collected during the latter half of November and during the first half of April. A summary of collections is presented in Table I. Table | collections affinis-alg. Number of putrida total Last half November December January February First half Dec.=-Mar. Of the six species taken during the December-March period two, D. transversa and nigromelanica, were represented by single specimens taken during December 1958. The other species were D. affinis- algonquin (180 flies), putrida (82 flies), robusta (26 flies), and tripunctata (13 flies). Five flies, dried beyond recognition, were not identified. They were apparently either affinis or the dark form of putrida. Species which were not taken during the critical period, but which were collected in November and/or April were the cosmo- politan species melanogaster, immigrans, hydei and busckii. All of these occurred in numbers of fifteen or less. The number of flies taken during the critical period of the two winters was disproportionate. In the winter 1957-58 only twenty flies were taken in thirteen collections from December through March. 82 Wallace D. Dawson During the same period of 1958-59 288 flies were taken in eighteen collections. The difference is probably due to the severity of the 1957-58 winter which was 3.8° F. colder per month than the mean average for Kentucky. A correlation was shown to exist both winters between the number of live flies collected and the temperature at the time of collection. Live flies were seldom obtained at temperatures below 40° F. In one instance a live D. putrida was in a trap taken at 18° F. No other live flies were taken at temperatures below 34° F. Occasionally dead flies were found in the traps and these were tabulated in the totals. In the winter 1957-58 fifteen of the twenty flies taken during the December- March period were alive. In the same period of 1958-59 241 of the 288 flies taken were living. During March and April 1958 twenty-eight flies of the D. affinis- algonquin sibling species were examined for sex. Twenty-one of these were females and six were males. In January through April 1959 sex ratio was determined in 334 affinis-algonquin. There were a total of 144 females and 190 males. Males outnumbered the females by ap- proximately twenty-five per cent during each of the four months. No significant correlation could be established between the number of flies or the relative abundance of any species obtained and precipita- tion or relative humidity. Collection of various types of material which were incubated in the laboratory to discern whether they served as suitable media for overwintering Drosophila eggs, pupae or larvae did not prove fruitful, although more than two hundred samples were collected in the course of the two winters. Special attention was given to decaying fruits, bark around slime fluxes, fleshy fungi, and similar materials which might be likely hibernacula. Direct Response Tests. A comprehensive series of experiments was conducted to determine the direct response of adult Drosophila after exposure to freezing and below freezing temperatures for various lengths of time. Including a period for feeding and acclimatization, all flies fell within an age range of ten to fifty-four hours from eclosion at the time they were placed into the freezer for testing. Relatively few of the flies were at the extremes of this age range. In these experiments the species D. affinis, robusta and melanogaster were used. These species were selected primarily because their seasonal population cycles are well known and they can be readily maintained in the laboratory. These species also presented the oppor- tunity to contrast two common native species with an abundant domestic species in a comparative temperature tolerance study. Studies of the Overwintering of Certain Drosophila Species 83 In each experiment of this series, except one, ten flies of each of three species were used. Each experiment was repeated once. In one test a total of seventy flies were used. Temperature ranges used were 0°, —5°, —10°, —15°, and —20° C. all +2°. Time exposures selected were one, four, eight, twelve, eighteen, twenty-four, thirty-six, forty-eight and sixty hours. These times and temperatures were selected because they best demonstrated the differences in survival capabilities of the species used. At the 0° C. range it was not necessary to use the shorter time exposures, since all Table II poUSU Species oe =5° -10° = =15° -20° Exposure affinis - 100 100 100 10 | robusta - 100 100 55 lor melanogaster - 100 95 55 10 affinis - 100 100 25 fe) 4 robusta ~ 100 90 O 6) melanogaster ~ 100 30 O re) affinis - 100 100 ¢) fe) 8 robusta - 100 60 e) - melanogaster - 90 5 8) = affinis - 95 90 0) - 12 robusta - Ons 50 ie) - melanogaster - fe) 5 6) - affinis 100 99 85 fe) = 18 robusta 100 100 fe) fe) = melanogaster 100 O 6) 0 - affinis - 90 10 O - 24 robusta = 60 @) - = melanogaster - 5 0 - - affinis - 95 ) - - 36 robusta - 50 ie) - - melanogaster - ie) 10) - = affinis 100 85 = - - 48 robusta 100 60 - - = melanogaster 40 6) - - - affinis 90 90 - - ~ 60 robusta 100 1S - - - melanogaster 5 0 - - = §4 Wallace D. Dawson species showed good survival at these levels. On the contrary at the lower temperature ranges, only short exposures were required since no flies survived beyond these limits. The results of this series of experiments are summarized in Table II. When a lethal dosage of fifty per cent for each species, as nearly as it could be determined from the results, is plotted against tempera- ture and time, the differential in survival at low temperature can be clearly demonstrated graphically (Fig. 1). D. affinis shows better 64 D. affinis 56 \ y \ eenemcenames Ds Fobtista 48 melanogaster 40 SZ Time tn Hours 24 16 eewreeene o° a -|0° =[5° =20° Temperature Centigrade Fig. 1. Lethal dosage fifty percent of Drosophila species exposed to low temperatures. survival at all levels indicated, with robusta somewhat intermediate between affinis and melanogaster. D. melanogaster shows only neg- ligible survival below —5° C for periods greater than twenty-four hours. Another experiment was conducted to determine approximately the nature of the survival response to freezing of several other species. In two separate experiments a total of fifty flies each of the species D. affinis, robusta, melanogaster, hydei, immigrans and cardini were used. D. hydei and immigrans are both cosmopolitan domestic species presumably of tropical or sub-tropical origin. D. cardini is a wild sub- tropical species of Florida and the West Indies. Thriving unaged flies were exposed to —5° C. for a period of eighteen hours. The results of this experiment indicate that hydei, immigrans and cardini are Studies of the Overwintering of Certain Drosophila Species 85 slightly less cold hardy than melanogaster and considerably less than affinis or robusta (Table III). Table Ill Percentage Actual Survival Complete 3 Survival Crippled Stunned Dead Survived Pertshed Species D. robusta De hydei tes ater