Transactions

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of the

Illinois

State Academy of Science

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Volume 64 No. I 1971 _

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JUN 2 19T1

NEW YORK BOTANICAL GARDEN1

9

TISAAH

TRANSACTIONS of the ILLINOIS STATE ACADEMY OF SCIENCE

Editorial Board :

Malcolm T. Jollie, Northern Illinois University, Editor and Chairman

Stanley H. Frost, Northern Illinois University, Geology Gordon C. Kresheck, Northern Illinois University, Chemistry John C. Shaffer, Northern Illinois University, Physics Darrell L. Lynch, Northern Illinois University, Microbiology Robert H. Mohlenbrock, Southern Illinois University, Botany

Articles in the Transactions pertaining to Biology, Chemistry, and Geology are abstracted in Biological Abstracts, Chemical Abstracts, and Abstracts of North American Geology.

The current Transactions may be obtained by payment of annual dues.

Exchanges may be arranged or previous issues may be purchased by addressing Paul Parmalee, Illinois State Museum, Springfield, Ill. 62706.

Mailing date, May 21, 1971.

TRANSACTIONS

OF THE

ILLINOIS STATE ACADEMY OF SCIENCE

VOLUME 64 - 1971

No. 1

Illinois State Academy of Science

AFFILIATED WITH THE

Illinois State Musetjm Division Springfield, Illinois

PRINTED BY AUTHORITY OF THE STATE OF ILLINOJ Richard B. Ogilvie, Governor

March 1, 1971

CONTENTS

On the Calculation of Association Constants of Polar Molecules.

By J. E. House, Jr., R. C. Reiter, and D. W. Hopkins . 3

Microclimate, Vegetation Cover, and Local Distribution of the Meadow Vole.

By Lowell L. Getz . 9

Association of Prodigiosin with Outer Cell Wall Components.

By Joseph C. Tsang and Dennis M. Kallvy . 22

Woody Vegetation of Hart Memorial Woods, Champaign County, Illinois.

By T. W. Root, J. W. Geis, and W. R. Boggess . 27

Growth Relationships between Aphelenchus avenae and Two Species of Nematopha- gous Fungi.

By H. L. Monoson . 38

Transverse and Crescent Cracks in Soybean Cotyledons Associated with Imbibition.

By Kuo-Chun Liu and A. J. Pappelis . 43

A Proposed Biochemical Mechanism of the Toxic Action of DDT.

By Robert C. Hiltibran . 46

Surfac Tensions of Binary Solutions of Nitroparaffins in Carbon Tetrachloride.

By Claude R. Gunter, Richard D. Madding, Jr., Thomas E. Hanson,

and Boris Musulin . 55

Computerized Curve Fitting: An Alternative to Graphical Interpretation.

By Boris Musulin . 67

On Various Methods Used in the Calculations of Inelastic Electron Atom and Elec¬ tron-Molecule Collisions.

By Yuh-Kang Pan . 84

NOTES

What Effect Does Prolonged Flooding Have on Ant Colonies?

By C. A. Dennis, D. Henry, D. Walch, G. Jones, and L. C. Shell . 95

Tribute to Dr. James W. Neckers.

By Richard T. Arnold . 96

A Convenient Method for the Preparation of Aromatic Diketones.

By Jerry Higgins and Joe F. Jones . 97

RE: Hawkins and Klimstra, “Deer Trapping Correlated with Weather Factors”. (Trans., 63:198-201).

By H. W. Norton . 99

Dr. Lester Wicks.

By Dr. Boris Musulin . 100

Professor Joseph Tykocinski Tykociner.

By Dr. Boris Musulin . 101

Dr. Byron Riegel President of American Chemical Society.

By Dr. Boris Musulin . 102

Biological Notes on Phloeotribus scabricollis (Hopkins) (Coleoptera: Scolytidale) .

By Milton W. Sanderson and James E. Appleby . 103

ON THE CALCULATION OF ASSOCIATION CONSTANTS

OF POLAR MOLECULES

J. E. HOUSE, JR, R. C. REITER, AND D. W. HOPKINS

Department of Chemistry, Illinois State University, Normal, Illinois 61761

Abstract. — A previously published method of calculating dipole association constants and dipole moments from dielec¬ tric constants of dilute solutions of polar substances in a nonpolar solvent is dis¬ cussed. A computer program is presented which enables simpler and more reliable treatment of the data. Some extensions of the previous method are presented and dis¬ cussed.

In 1964, Treiner, Skinner, and Fuoss published a method of evalu¬ ating self-association constants and dipole moments of polar molecules in nonpolar solvents. The method in¬ volves a graphical treatment of di¬ electric constants and concentrations of dilute solutions of the polar sub¬ stances. Further, the method as¬ sumes antiparallel dimerization of the dipoles which results in a can¬ cellation of the contributions which the permanent dipole moments make to the dielectric constants of the solutions. The method depends upon the fact that when dipole associa¬ tion occurs, the total polarization in¬ creases less rapidly than when there is no such association. Thus, the rate of increase of dielectric constant with increasing concentration is in¬ dicative of the extent of solute as¬ sociation.

In the calculations, a rather com¬ plicated function of dielectric con¬ stant and concentration, G (c, c) and derived and its reciprocal is plotted

against the product of G (e, c) and c. The slope and intercept of the resulting plot can be used to calcu¬ late the association constant and di¬ pole moment of the solute (Treiner, et al., 1964). The Debye-Clausius- Mosotti approximation of volume polarization was used and the vol¬ ume polarization of the dimer was assumed to be twice that of the monomer.

The work described in this paper was undertaken with two main ob¬ jectives. First, a computer program was developed to carry out the tedi¬ ous, repeated evaluation of the func¬ tion G (e, c). Because the calculat¬ ed values of the association constant and dipole moment are sensitive to small errors in the function G (e, c), least squares evaluation of the values of the slopes and intercepts is in¬ corporated into the program.

Second, instead of assuming that the volume polarization of the dim¬ er is twice that of the monomer, the computer program allows a variable factor to be introduced and its ef¬ fect on dipole moment and associa¬ tion constant to be determined. Per¬ haps this factor should vary as an inverse function of the strength of the forces of molecular association. The results of these extensions of the dipole association theory of Treiner, Skinner, and Fuoss are pre¬ sented and discussed.

[3]

4

Transactions Illinois Academy of Science

Calculations

The calculations were carried out using an IBM 360/40 computer with the program written in FORTRAN IV. The program and some general comments on its use follow. Comments :

1. The “Fuoss G” function,

1.189 x lO -X (E - Es)

Es + 2 J[_C (E + 2)

3RHOs0iv

(Es - 1) (/3 - Mwt/1000)

can be reproduced from the pro¬ gram by substitution of the in¬ termediate function F of S. 13 into the equation in S. 14, with Z = 2.

2. The notation X (I) represents the value of property X for the “Ith” solution. Other previous¬ ly undefined key symbols and their meanings follow.

E = e = dielectric constant of the solution

ES = Es = dielectric constant of the solvent

RHOSoiv = solvent density

BETA = /3 = a factor relating solvent and solution densities as RHO = RHOSoiv + PC. (Densities varied linearly in the range used.)

WMOL = Mwt = molecular weight of solute

NPTS = number of data pairs to be plotted

ALPIO = a10 = solute mono¬ mer electronic polarization

Notation in the “ least squares” segment of the program is stand¬

ard, with “s” equivalent to

3. The output format will result in clearly labeled results. The data read by S.05 can be any identi¬ fying material, such as solute name.

4. The program recycles until a blank card is read by S.02, the parameter read instruction.

5. As listed here, the program varies Z from 1.00 to 2.50 in increments of 0.25. Removal of S.09, S.10 and S.39, and replacement of S.ll by Z = 2. will elminate this procedure.

6. The factor 298.2 in S.30 is tem¬ perature (°K) and can be chang¬ ed depending on the data used.

0001 DIMENSION C(15),E (15),G(15)

0002 1 READ (1,2) NPTS, ES, BETA, WMOL, RHO, ALPIO

0003 2 FORMAT ( 12, 2X,F5.3,2X, F5.4,2X,F6.2,2X, F7.5,2X,E14.8)

0004 IF(NPTS)3,99,3

0005 3 READ (1,4)

0006 40 FORMAT (80H

1 )

0007 READ (1,5) (C(I),E(I), 1=1, NPTS)

0008 5 FORMAT (F7.6, IX, F6.4) 0009 DO 9 II — 1,7 0010 AI=II 0011 Z=.75+.25*AI

0012 DO 6 1=1, NPTS 0013 F=( (1.1890E-21) / (ES+

2.) )*( (E(I)-ES)/ (E(I) +2.)-(ES-l.)*

1 ( BET A- WMOL /1000. ) * C(I)/(3.*RHO))

House et al — Association Constants

5

0014 6G(I)=F/C(I)-(Z*

ALP10) /2.

C LEAST SQUARES FIT FOR PLOT OF 1/G VS CG FOLLOWS 0015 SX=0.

0016 SXY=0.

0017 SY=0.

0018 SXX=0.

0019 DO 7 I=1,NPTS 0020 X=C(I)#G(I)

0021 Y— l./G(I)

0022 WRITE ( 3,50 ) X,Y

0023 50 FORMAT (1H0,2E14.7 0024 SX=SX+X 0025 SXY=SXY+X*Y 0026 SY=SY+Y 0027 7 SXX=SXX+X**2 0028 SLOPE= ( SX*SY-NPTS

*SXY) / (SX*SX-NPTS* SXX)

0029 CEPT=(SXY*SX-SY* SXX) /(SX*SX-NPTS* SXX)

0030 DEB YE=SQRT ( 3 *1.3805 E-16*298.2/CEPT)*1.E+ 18

0031 AKEQ=SLOPE/(2.#

CEPT#CEPT)

0032 WRITE (3,14)

0033 14 FORMAT (//////80H

1 )

0034 WRITE (3,4)

0035 WRITE (3,20) Z

0036 20 FORMAT ( 1H0,4HZ = ,F5.2)

0037 WRITE (3,8) CEPT,

SLOPE, DEBYE, AKEQ

0038 8 FORMAT (1HO, ^IN¬ TERCEPT = ,E14.8/9H SLOPE = ,E14.8/17H DIPOLE MOME INT = ,F6.2/24H ASSOCIATION CONSTANT = ,E14.8)

0039 9 CONTINUE 0040 GO TO 1 0041 99 STOP 0042 END

SAMPLE INPUT

.16260 3.203 .10880 2.870

.06239 2.595 .04128 2.471 .03021 2.405

.01999 2.349 .07273 2.660 .05348 2.545 .03838 2.456 .02492 2.379

PNITRO ANILINE 10 2.233 .0406 138.12 1.02796 .15000000E-22

SAMPLE OUTPUT

X

0.9264186E-23

0.1390161E-22

0.1906553E-22

0.2544358E-22

0.7411161E-23

0.1084541E-22

0.1478510E-22

0.2187612E-22

0.3629993E-22

0.5170795E-22

Y

0,2689928E 22 0.2760829E 22 0.2805061E 22 0.2858479E 22 0.2697282E 22 0.2785510E 22 0.2792000E 22 0.2851968E 22 0.2997250E 22 0.3144584E 22

PNITRO ANILINE Z = 2.00

INTERCEPT = 0.26292032E 22 SLOPE = 0.99281275E 43 DIPOLE MOMENT = 6.85

ASSOCIATION CONSTANT = O.71810728E 00

Discussion

Dielectric constant, density, and concentration data of Treiner, Skin¬ ner, and Fuoss (1964) for solutions of p-nitroaniline (PNA), of m-nitro- phenol (MNP), and of pyridinium dicyanomethylide (PDM) in dioxane were used in the computer calcula¬ tions. A comparison of the results of

6

Transactions Illinois Academy of Science

these workers and the results ob¬ tained in this work is shown in Ta¬ ble 1. The results of the calculations for solutions of PNA, for which Treiner, et al., provide the most data, show almost identical values for the association constants. The small dif¬ ferences are due to the use of a least squares routine for obtaining the slope and intercept of the plot of cG(c, c) against 1/G(e, c). The val¬ ues of these functions were reported by Treiner, et al., for PNA solutions and the values obtained using the techniques described in this work agree closely.

For the calculations on MNP and PDM solutions the agreement is not nearly as good. In the case of MNP, the association constant calculated by Treiner, et al., is too high, as is the dipole moment. For PDM, the agreement between the dipole mo¬ ments is good but the association constants differ greatly.

Since the value of the dipole mo¬ ment is derived from the intercept of the cG(e,c) vs 1/G(c,c) plot, it is obvious that the major errors in the calculations of Treiner, et al., are in

the intercept for MNP and the slope for PDM. These results show the ex¬ treme sensitivity of the calculations to small errors in the determinations of the slopes and intercepts of the plots of cG (e,c) against 1/G(c,c). Because of the small variations in these functions in some cases, it would appear that reliable results are not likely to be obtained unless very accurate data are used and the calculations carried out by means other than graphical methods. How¬ ever, it appears that in the case of MNP, the intercept is so much in error that the error must be caused by a systematic numerical error in carrying out the calculations. In fact, if in the equation giving G(e,c) (Es + 2) is replaced by (Es-j-l) the result is an error in G(e,c) which would lead to the results published by Treiner, et al.

Intuitively, it seems quite likely that the volume polarization of the dimer should not be exactly twice that of the monomer because of mu¬ tual inductive effects. Consequently, this assumption was tested by allow¬ ing the volume polarization of the

Table 1. — Calculated Association Constants and Dipole Moments for PNA, PDM, and MNP.

Compound

K Assoc.,

(1/ mole.)

Slope

x 10 43

Intercept x 10-21

TSFa

This

Workb

TSFa

This

Workb

TSFa

This

Workb

TSFa

This

Workb

PNA .

6.91

6.85

0.8

0.72

1.06

0.99

2.59

2.63

PDM .

9.2

9.30

3

1.34

1.28

0.55

1.46

1.43

MNP .

4.38

3.75

0.37

0.26

3.10

3.92

6.43

8.76

a Average values obtained from a graph, dipole moments, and association contants published by Treiner, et al., (1964) when used in the equations 1018^ = \/3KT/ Intercept and K = slope/2

( Intercept) 2.

b Values obtained using densities and dielectric constants published by Treiner, et al., when the computer method is used.

House et al — Association Constants

7

dimer to vary from 1.0 to 2.5 times that of the monomer. The results of these calculations are shown in Table 2. It is immediately obvious that the calculated values of the dipole mo¬ ment and association constant are rather insensitive to the variable fac¬ tor, Z. Taking the volume polariza¬ tion of the dimer to be the same ,as that of the monomer or taking it to be 2.5 times that of the monomer results in a difference in the calcu¬ lated value of the association con¬ stant of only a few per cent. Because

of the introduction of relatively large errors when graphical methods are used in the calculations, the effect of varying the factor would probably be unclear or unnoticed unless the calculations were carried out by means of a computer.

Throughout the calculations it be¬ came increasingly clear that the most critical quantity in evaluating the function G(e,c) is (E0-Es). When the dielectric constant of the solution differs very little from that of the solvent, this quantity approaches

Table 2. — Calculated Values of Association Constants and Dipole Moments Showing the Effect of the Z Factor.

Data for PNA

z

10 22 x Intercept

10 43 x Slope

Dipole Moment

K

1.00

0.2579

0.9285

6.92

0.6979

1.25

0.2592

0.9441

6.90

0.7029

1.50

0.2604

0.9600

6.89

0.7079

1.75

0.2617

0.9762

6.87

0.7130

2.00

0.2629

0.9928

6.85

0.7181

2.25

0.2642

1.010

6.84

0.7234

2.50

0.2655

1.027

6.82

0.7287

Data for MNP

1.00

0.8259

0.03252

3.87

0.2384

1.25

0.8379

0.03405

3.84

0.2424

1.50

0.8504

0.03567

3.81

0.2466

1.75

0.8632

0.03740

3.78

0.2509

2.00

0.8764

0.03924

3.75

0.2554

2.25

0.8901

0.04121

3.72

0.2601

2.50

0.9041

0.04330

3.70

0.2649

Data for PDM

1.00

0.1412

0.5287

9.35

1.325

1.25

0.1417

0.5334

9.34

1.329

1.50

0.1421

0.5382

9.32

1.334

1.75

0.1425

0.5428

9.31

1.337

2.00

0.1429

0.5473

9.30

1.341

2.25

0.1433

0.5520

9.28

1.345

2.50

0.1437

0.5567

9.27

1.349

8

Transactions Illinois Academy of Science

zero. Thus, subtraction of «10 can result in a G(e,c) which is about zero or even a negative quantity. This can result in a negative value for the association constant, which has no physical significance.

Treiner, et al have stated that the energy of interaction of a pair of di¬ poles varies as the square of the dipole moment,

K « lO'3 Na3 e“u/kT

where N is Avogadro ’s number, a3 is the volume occupied by a pair of dipoles, and u is the energy of inter¬ action. Therefore, a linear relation¬ ship should exist between log K and /*2. Their report presents such a relationship. In view of the errors in the values they reported for the

association constants, it appears somewhat fortuitous that the linear relationship was obtained. The pres¬ ent results do not enable one to say with certainty that the relationship is linear.

ACKNOWLEDGMENT

The authors would like to acknowl¬ edge the support and cooperation of the Computer Services of Illinois State University.

Literature Cited

Treiner, C., J. F. Skinner, and R. M. Fuoss. 1964. Dipole Association. J. Phys. Chem., 68, 3406-3409.

Manuscript received June 21, 1970

MICROCLIMATE, VEGETATION COVER, AND LOCAL DISTRIBUTION OF THE MEADOW VOLE

LOWELL L. GETZ

Department of Zoology, University of Wisconsin, Madison, 53706

Abstract. — A comparison was made of the microclimate of the meadow vole, Microtus pennsylvanicus, in situations with varying amounts of cover. Substrate and surface air temperatures were higher in areas with sparse cover (not utilized by meadow voles) than in optimal habitats with dense cover. Relative humidities were higher in the optimal habitats than in situations not inhabited. There was no correlation between absolute humidity and amount of vegetation cover. Only the higher temperatures in areas with sparse vegetation have the potential of causing the voles to avoid these sites. Relative and absolute humidities were not low enough in the sites with sparse cover to place a serious physiological stress on the voles. Humidity does not appear to play a signifi¬ cant role in the avoidance of areas with sparse vegetation by the meadow vole.

Introduction

The local distribution of the mea¬ dow vole, Microtus pennsylvanicus , has been correlated with moisture (Getz, 1961, 1963; Lantz, 1907; Findley, 1954; DeCoursey, 1957) and amount of vegetation cover (Getz, 1961, 1970b; Pearson, 1959; Mossman, 1955 ; Eadie, 1953 ; Zim¬ merman, 1965). The actual factors involved in these correlations have not been determined, however; mic¬ roclimate, especially humidity, has been suggested as a possible factor (Getz, 1963).

Data comparing the microclimate of moist and dry graminoid situa¬ tions (Getz, 1965, 1970a) indicate

that humidity differences between such situations are slight. Humidity apparently is not responsible for higher population densities of the meadow vole in moist marshes. The above studies compared situations in which there was dense cover in both the moist and dry habitats. Micro¬ climate data at the level of vole run¬ ways are not available from grami¬ noid sites with varying densities of cover. The data presented in a prior paper (Getz, 1970b) which compared areas with dense and sparse cover were all taken in a wet marsh. Hu¬ midity differences would not be ex¬ pected to be great in areas with sat¬ urated substrates.

During the summer of 1968 data were obtained in southern Wisconsin which aid in evaluating the signifi¬ cance of microclimate on the local distribution of the meadow vole. These data were obtained in an area with varying amounts of vegetation cover ; the sites included those deem¬ ed optimal for the meadow vole as well as sites which the meadow vole did not utilize (Getz, 1970a).

Description of Study Areas

All work was conducted in the University of Wisconsin Arboretum, Madison, Wisconsin. Five stations, all in the southern part of the Grady Tract, were selected for the main

[9]

10

Transactions Illinois Academy of Science

study ; other stations in this tract and elsewhere in the Arboretum were spot-checked for comparison. The five main stations were all within 100 m of each other.

Station 1 was in a stand of Agro- pyron repens. The vegetation was 30-40 cm tall ; litter formed a dense mat over the surface. Light penetra¬ tion through the vegetation was only 0.02% (light penetration was used as an index of amount of cover pres¬ ent; see below and Table 1. Station 2 was in blue-grass ( Poa pratensis) which formed a dense mat 20-30 cm above the surface. The total cover was less than that of Station 1 (1.1% light penetration). Topographically, both Stations 1 and 2 were 20-30 cm higher than the other 3 stations. Station 3 was in a low marsh which supported a pure stand of Car ex lanuginosa. The vegetation at this station was 40-50 cm tall and had a cover value similar to that of Station 2 (Table I). Station 4 was in an area supporting a relatively sparse growth of various species of grasses and forbs. The average height of

the vegetation was 30 cm and the cover approximately l/10th that of Stations 2 and 3. Station 5 was in an area supporting a very sparse growth of several species of grasses and forbs. The height of the vege¬ tation was 15-20 cm. The cover was so sparse that the surface was rela¬ tively exposed in most places ; light penetration was high (17.5%).

In addition to the main study area, various other sites were spot-checked. One included a mowed blue-grass area. There was a very dense growth of grass in this area ; the surface was completely covered with a mat of green vegetation. Vegetation height was only 2-5 cm at the site studied. The microclimate measurements were therefore taken in the “crown” of the vegetation, i.e., within the green leaves. Spot-checks were also made under a stand of young oak ( Quer - cus spp.) shrubs (2 m tall). Crown coverage of the oaks in this site was 95-100% ; there was no understory of grass or forbs. The measurements were made over leaf litter 2-3 cm thick.

Table 1. — Soil temperature, soil moisture, and light penetration at the main study stations. See text for techniques.

Station

Mowed

Date

1

2

3

4

5

Blue-

Grass

Shrubs

Soil temp. (C) .

Surface .

2 Aug .

24.6

26.8

26.8

29.3

30.4

32.8

25.5

— 7 cm .

20.8

22.4

21.4

24.1

25.9

29.5

21.8

Soil moisture

g/280 cm3 .

25 July. . . .

112

116

142

163

153

108

103

2 Aug .

102

107

133

147

139

55

68

% Saturation .

25 Tuly .

35.4

34.7

42.6

50 1

43.7

26.0

29.1

2 Aug .

30.8

31.5

40.2

46.5

39.7

13.4

22.5

Per cent light

penetration1 .

2 Aug .

0.02

1.1

1.0

10.5

17.5

61.0

1 % of full sunlight.

Getz — Microclimate of Vole

11

Some data were obtained from a dry south-facing hillside which sup¬ ported a moderately dense stand of bluegrass ; the vegetation was 20-30 cm tall and formed a less dense cover (4.1% light penetration) than that at Station 2. Comparative data were taken in an adjacent site (2 m dis¬ tance) which was mowed. The vege¬ tation was much more sparse in this area than in the mowed area' de¬ scribed above ; the surface was al¬ most entirely exposed (47.5% light penetration) .

A few measurements were also taken in a tail-sedge {Car ex sp.) marsh, the typical habitat of the meadow vole in this region, for com¬ parison with the drier sites. The sedges grew in clones approximately 10-20 cm in diameter at the base and 1.0-1. 5 m tall. The bases of the clones were 30-50 cm apart ; the crown coverage was 100%, however. Light penetration was 1-2%.

Observations of vole sign and live- trapping were conducted in the above described vegetation types during the summers of 1967 and 1968. These indicated that Station 3 and the tail- sedge marsh area were the optimal habitats of the meadow vole (Getz, 1970a). Late in the summer of 1968 the vole population had also extend¬ ed out into areas including Stations 1, 2, 4 and the unmowed hillside. Station 5, the two mowed areas, and the shrub area were not utilized by the meadow vole. These latter areas are not considered suitable habitats for this species.

Methods

A series of microclimate measure¬ ments (1 cm above the substrate sur¬ face) were made at approximately 2-hour intervals between 0745 and 1900 on 2 August 1968. Another series were made between 1430 and

2045 on 22 July 1968. Data were not obtained during the night since these observations and other spot- checks indicated all stations had sim¬ ilar temperature and relative humid¬ ities between 2000 and 0800 ; relative humidity was 95-100% during this latter period. Five other series of spot-checks of all stations were made during July and August. All meas¬ urements were made on clear days at least 3 days after the last rain.

Surface microclimate. — A ther¬ mistor psychrometer was used to ob¬ tain dry-bulb and wet-bulb readings at the desired sites. These data gave surface air temperatures and were used to calculate relative and abso¬ lute humidities.

Three sites (within a 10 m radius) were sampled at each station ; three sets of dry and wet-bulb readings were taken at each site (all within aim radius) ; there were therefore nine sets of readings from each sta¬ tion each time it was checked.

Measurements were taken 1 cm above the surface. The barrel of the psychrometer was inserted through the vegetation so as to keep to a minimum disturbance to the crown cover and litter layer. All measure¬ ments were taken at the exact same place during each series of checks. The hole in the litter was closed after each set of measurements was com¬ pleted. One set of dry and wet bulb readings was also taken 1 m above the surface at each site. These pro¬ vided comparison of above-surface air temperatures at the various sta¬ tions.

The stations were visited in the same sequence ; it took 35-40 min to make the complete set of measure¬ ments. When other stations were spot-checked, all were completed within 30 min after the measure¬ ments at the regular stations were finished. See also Getz (1970a) for

12

Transactions Illinois Academy of Science

other microclimate data obtained in the same study area.

Soil temperature. — A Yellow- springs telethermometer with an at¬ tached hypodermic thermistor probe was used to measure substrate tem¬ peratures. Substrate temperatures were measured at the same number of sites as described above. Three sets of surface (sensitive portion of the probe only touching the surface) and subsurface (7 cm below the sur¬ face) temperature readings were tak¬ en at each site.

Soil moisture. — a gravimetric method (Getz, 1970a) was used to estimate substrate moisture ; data were obtained both in terms of per cent saturation and g water/280 cm3 of substrate. One 280 cm3 plug of substrate was taken at each station. Two sets of samples were taken, 25 July and 2 August 1968.

Light penetration. — Amount of light penetration through the vege¬ tation was used as an index of total vegetation cover at each station. A previously described device (Getz, 1968) was used to measure light in¬ tensity above the vegetation and at the surface of the substrate. This device creates a minimum of dis¬ turbance to the vegetation crown and litter. One above-surface reading and 12 surface readings were tak¬ en at each station. The two most aberrant surface readings at each station were omitted from the cal¬ culations. Readings were taken 1100- 1200 on a cloudless day.

Results

Main Study Area

Substrate moisture. — There were frequent showers in southern Wis¬ consin during the summer of 1968. As a result, substrate moisture was relatively high at all stations. There was no consistent correlation between

amount of vegetation cover and sub¬ strate moisture (Table 1). In gen¬ eral the two areas with less vegeta¬ tion had higher substrate moisture while those with the greatest amount of cover were drier. The former sta¬ tions were slightly lower topograph¬ ically than the latter two. Al¬ though Station 3 was the lowest of the five, the substrate moisture of this station was intermediate be¬ tween that at the other stations. The relatively slight microrelief differ¬ ences (combined with the frequent rains) appeared to be as important as did the amount of vegetation cover in influencing substrate moisture.

Substrate temperature. — Both sur¬ face and subsurface temperatures tended to be correlated with the amount of vegetation cover at each site; mid-day temperatures were higher and diel fluctuations greater where the vegetation was sparse. The maximum difference in substrate sur¬ face temperatures between stations was 5.8 C ; maximum subsurface dif¬ ference was 5.1 C (Table 1). These differences are essentially the same as those observed in the surface air temperatures (see below) . Although substrate temperatures were not measured during the night, there would be less difference in substrate temperature during these times than during mid-day. Substrate tempera¬ tures would also vary little on over¬ cast days.

Surface air temperatures. — Air temperatures 1 m above the surface did not differ significantly between any of the various stations studied. Wind currents apparently caused enough mixing of air to prevent de¬ velopment of different temperature profiles at each station.

As might be expected, surface air temperatures in general directly re¬ flected the amount of vegetation cover; higher daytime temperatures

Getz — Microclimate of Vole

13

and greater diel fluctuations occur¬ red at the sites with lesser cover than at those with greater cover. Differ¬ ences in temperatures between the five stations occurred only during

the period of 1000 to 1600. The max¬ imum recorded difference was 7.5 C; observed differences were normally less than this, however (Figs. 1, 2; Table 2).

Table 2. — Spot checks of air temperatures (C) 1 cm above the surface; values represent an average of 9 readings at each station.

Station

Mowed

Date

Time

1

2

3

4

5

Blue-

Grass

Shrubs

21 July .

1420-1510

28.5

29.4

29.6

29.9

30.0

28.9

28.2

23 Tuly .

0710-0750

14.8

15.2

14.6

15.2

15.6

16.2

14.9

25 July .

1420-1450

22.4

23.2

25.4

26.9

27.3

28.4

24.9

29 Tuly .

1350-1430

22.1

25.0

22.5

25.8

26.8

24.9

23.7

26 August .

1430-1540

18.6

20.3

18.7

19.7

24.0

24.1

20.3

Figure 1. Air temperatures 1 cm above the surface, 2 August 1968.

14

Transactions Illinois Academy of Science

|0— 1 - 1 - 1 - 1 - 1 - 1 - 1 - 1 - r

14 15 16 17 18 19 20 21

Time of Day

Figure 2. Air temperatures 1 cm above the surface, 22 July 1968.

Spot checks revealed that at night and on overcast days there was es¬ sentially no difference in surface air temperatures between the stations with sparse cover and those with con¬ siderable cover.

Relative humidity. — There was no significant difference in relative humidity 1 m above the surface be¬ tween any of the stations studied.

Major differences in surface rela¬ tive humidity between the five sta¬ tions existed only during the period of 1000-2000 ; at other times differ¬ ences were less than 5% (Fig. 2; Table 3). Between 2000 and 0800 relative humidity was approximate¬ ly 95% at all stations.

There appeared to be more similar¬ ities between relative humidity and the amount of cover than between relative humidity and substrate mois¬ ture (Figs. 3, 4; Tables 1 & 3.) The lowest relative humidities were observed where cover was sparse and the highest where there was the most cover.

The maximum difference in rela¬ tive humidity between the station with the sparsest cover and that with the most dense cover was 20%. The maximum difference between other stations was only 11%, however. Even at the stations with sparse cover, relative humidity seldom went below 60% ; normally humidities

Getz — Microclimate of Vole

15

Table 3. — Spot checks of relative humidity {%) 1 cm above the surface; values represent average of 9 readings at each station.

Station

Mowed

Date

Time

1

2

3

4

5

Blue-

Grass

Shrubs

21 Tuly .

1420-1510

82.7

75.7

71.2

67.8

62.7

81.4

70.5

23 July .

0710-0750

98.3

97.3

97.0

95.7

95.7

93.7

94.0

25 July .

1420-1450

90.0

90.8

81.7

87.0

75.2

90.8

79.3

29 Tuly .

1350-1430

84.8

80.9

77.9

80.2

71.1

93.0

69.7

26 August .

1430-1540

77.6

76.1

73.9

79.8

67.1

87.0

59.7

Time of Day

Figure 3. Relative humidities 1 cm above the surface, 2 August 1968.

were at least 70%. It therefore ap¬ pears that differences in relative hu¬ midity between optimal habitats and situations where the meadow vole does not occur are not great.

Absolute humidity. — Absolute humidities were highest during the

day. During the night and on over¬ cast, humid or rainy days the air at the surface was essentially saturated regardless of the amount of vegeta¬ tion cover. Absolute humidity at these times was essentially a func¬ tion of surface air temperature. Ab-

16 Transactions Illinois Academy of Science

Time of Day

Figure 4. Relative humidities 1 cm above the surface, 22 July 1968.

solute humidity differences between stations existed only during the pe¬ riod of 0900 to 1900. The maximum difference between any two stations was 5.8 mg*/l normally the differ¬ ences were much less than this (Figs. 5, 6; Table 4) .

Although there was no distinct correlation between absolute humid¬ ity and amount of vegetation cover, the higher absolute humidities fre¬ quently occurred at the stations with lesser cover. This is a reflection of

the significance of temperature on absolute humidity; the higher mid¬ day surface air temperatures (and thus the greater capacity for the air to hold water) in the area with less cover more than compensated for the lower relative humidity at these sites.

Other Areas

Mowed blue grass. — Air tempera¬ tures 1 cm above the surface were not significantly higher than those

Getz — Microclimate of Vole

17

Table 4. — Spot checks of absolute humidity (mg/1) 1 cm above the surface; values represent average of 9 readings at each station.

Date

Time

1

2

Station

3

4

5

Mowed

Blue-

Grass

Shrubs

21 July .

1420-1510

22.5

25.5

22.5

26.6

25.6

29.8

20.3

23 July .

0710-0750

13.4

13.5

13.1

13.1

13.4

13.6

12.7

25 July .

1420-1450

19.7

18.8

21.7

25.5

22.3

22.0

16.5

29 July .

1350-1430

18.2

21.1

17.5

21.9

20.3

23.9

16.3

26 August .

1430-1540

13.1

14.4

13.0

14.7

16.3

21.1

11.5

Figure 5. Absolute humidities 1 cm above the surface, 2 August 1968.

18

Transactions Illinois Academy of Science

at other stations with more cover (Tables 1 and 2). Since measure¬ ments taken 1 cm above the surface were essentially in the vegetation “crown”, transpiration cooling may be responsible for the lower air tem¬ peratures.

Relative humidity 1 cm above the surface was normally as high as or higher than (up to 10% higher) that at any station with dense cover (Ta¬ ble 3). The measurements were probably high owing to moisture re¬ leased via transpiration. The abso¬ lute humidities in the mowed area were normally higher than those at

the stations with the most cover (Table 4).

Shrub area. — Surface air temper¬ atures were lower than most ofRhe stations with graminoid cover (nor¬ mally temperatures at the former were intermediate between the ex¬ tremes of the five main stations). The crown cover of the shrubs shad¬ ed the area sufficiently to modify surface temperatures (Table 2).

Surface relative humidity was es¬ sentially the same as that recorded at Station 5 (Table 3). The com¬ bined low relative humidity and low temperatures resulted in lower ab-

Getz — Microclimate of Vole

19

solute humidities in the shrub area than were normally recorded in the graminoid areas (Table 4) .

Dry hillside. — In general the sur¬ face air temperatures in the unmow¬ ed site were higher than at any of the five main stations. The relative humidities were intermediate to those observed at the other sites (except for the 26 August reading) ; absolute humidities were similar to those at the five main stations (Table 5).

Surface air temperatures were up to 4.2 C higher and relative humidi¬ ties as much as 21% lower in the mowed site than in the unmowed area. Absolute humidities were nor¬ mally 2-5 mg/1 higher in the mowed area. The differences from the other mowed area undoubtedly resulted from the more sparse growth of vegetation in the hillside area. When the grass was mowed, the re¬ maining vegetation was so sparse that transpiration was not as sig¬ nificant a factor in the microcli¬ mate as it was in the other mowed area.

Marsh. — Although only 3 spot- checks were made of microclimate at this site, they did not indicate sig¬ nificant differences from drier sites with dense vegetation (Table 5). In general, relative and absolute hu¬

midities were actually slightly less than at sites with dense cover. The green part of the leaves in this area were some distance above the sur¬ face (the parts close to the surface were dried out) so that there was little transpiration near the surface.

The more open character of the vegetation permitted drying of the soil surface ; this apparently re¬ duced evaporation from the surface. The humidity at the surface in this type of marsh vegetation is therefore lower than that at drier sites where there is dense, but low, vegetation cover. In any event, air humidity near the surface was not higher in this type of marsh (op¬ timal habitat for the meadow vole) than in drier upland sites (marginal habitats of the meadow vole).

Discussion

As would be expected there was in general an inverse relationship between amount of vegetation cover and substrate and surface air tem¬ peratures. The greater the cover, the lesser the extremes in substrate and surface air temperatures. Relative humidity was more positively related to vegetation cover ; sites with more cover had higher relative humidities.

Table 5. — Surface air temperatures and relative (RH) and absolute (AH) humidities at other sites in the vicinity of the main study area. Values represent averages of 9 readings at each site.

Date

Time

Mowed Hillside

Unmowed Hillside

Marsh

Temp.

(C)

RH

%

AH

mg/1

Temp.

(C)

RH

%

AH

mg/1

Temp.

(C)

RH

%

AH

mg/1

25 July .

1500

30.1

70.8

22.4

27.9

80.9

19.5

29 July .

1445

28.2

62.4

19.2

25.7

79.0

21.3

2 August .

1000

26.4

74.6

15.3

22.2

92.0

20.0

22.2

86.0

18.5

2 August .

1620

27.9

55.9

17.9

25.6

77.2

20.1

22.5

80.0

19.7

26 August .

1610

26.4

50.7

14.3

23.2

58.5

13.5

21.7

74.6

15.9

20

Transactions Illinois Academy of Science

Owing to the higher air tempera¬ tures in sites with less cover, there was no direct relationship between cover and absolute humidities. The absolute humidity in the sites with the least cover were normally slight¬ ly higher than in those with the most cover, however.

The magnitudes of difference be¬ tween the microclimate of the vari¬ ous sites included in the present study were relatively small. The vegetation types studied ranged from optimal habitats to those not inhab¬ ited by the meadow vole ; the micro¬ climate of the optimal habitats of this species is therefore only slightly different from that of other vegetat¬ ed situations not normally util¬ ized. Even when sites such as closely mowed areas are considered, microclimates do not differ greatly from situations in which the meadow vole does occur. In close-clipped, dense vegetation the microclimate actually may be similar to that in optimal habitats.

Of the factors studied, air and substrate temperatures varied the most. The 5 to 7 C differences that were recorded may be sufficient to make the sparsely vegetated sites un¬ suitable for the meadow vole. Data pertaining to the temperature toler¬ ances and preferences of the meadow vole are not available, however.

Relative humidity differences be¬ tween the various cover types were at the most 20% and normally in the order of only 10%. These differ¬ ences do not appear to be great enough for relative humidity to place a physiological stress on the voles, if they were to attempt to inhabit situa¬ tions with sparse cover. Although the water requirements of the mea¬ dow vole vary with relative humid¬ ity, such requirements are signifi¬ cantly higher only at relative humid¬

ities of less than 50% (Getz, 1963). The lowest humidity recorded in the present study was above 50% (nor¬ mally they were above 70%).

Differences in microclimates oc¬ curred only for about a 9-10 hr pe¬ riod during mid-day, and then only on clear days. On overcast days and from 2000 to 1000 there was essen¬ tially no difference in the microcli¬ mate between sparse-cover and dense- cover situations. Any stress placed on the meadow vole by microclimatic differences would therefore be fur¬ ther reduced. It is possible, however, that the higher temperatures during mid-day on clear days would be suf¬ ficient to keep the voles from inhab¬ iting areas with sparse cover.

Another study in same area ( Getz, 1970a) also failed to find sufficient microclimate differences, humidity in particular, to account for the local distributional pattern of the meadow vole.

The results of this study and pre¬ vious ones (Getz, 1965; 1970a, 1970b) make it rather obvious that relative humidity is not a significant factor in the local distribution of the meadow vole. This applies to situa¬ tions involving varying amounts of cover as well as moist and dry habi¬ tats. The magnitude of habitat dif¬ ferences and the amount of time such differences do exist are not sufficient to place a significant stress on the voles.

Studies of temperature tolerances and preferences are required to es¬ timate the actual significance of tem¬ perature on the local distribution of the meadow vole. It is probable that other factors such as behavioral preferences, and predation (in the case of varying cover; Getz, 1970b) may also be at least partly responsi¬ ble for the association of the mea¬ dow vole with dense vegetation.

Getz — Microclimate of Vole

21

Acknowledgments

This study is a part of project number 68-184 of the University of Wisconsin Ar¬ boretum; it was supported by NSF GB- 6203. Miss Melinda A. Novak assisted with the field work and analysis of the data. A contribution from the Biological Sciences Group, University of Connecticut, Storrs. Present address: Department of Zoology, University of Illinois, Urbana, 61801.

References

DeCoursey, G. E. 1957. Identification, ecology, and reproduction of Microtus in Ohio. J. Mamm., 38: 44-52.

Eadie, W. R. 1953. Response of Micro¬ tus to vegetation cover. J. Mamm., 34: 263-264.

Findley, J. S. 1954. Competition as a possible limiting factor in the distribution of Microtus. Ecology, 35 : 418-420. Getz, L. L. 1961. Factors influencing the local distribution of Microtus and Syn- aptomys in southern Michigan. Ecology, 42: 110-119.

- . 1963. A comparison of

the water balance of the prairie and meadow voles. Ecology, 44: 202-207.

- . 1965. Humidities in vole

runways. Ecology, 46: 548-550.

- . 1968. A method for

measuring light intensity under dense vegetation. Ecology, 49: 1168-1169.

- . 1970a. Habitat of the

meadow vole during a “population low.” Amer. Midi. Nat., 83: 455-461.

- . 1970b. Influence of vege¬ tation on the local distribution of the meadow vole in southern Wisconsin. Occ. Paps, in Biol., Univ. Conn., 1: 213-241.

Lantz, D. E. 1907. An economic study of field mice (genus Microtus). U.S. Dept. Agr. Biol. Surv. Bull., 31: 1-64.

Mossman, A. A. 1955. Light pentration in relation to small mammal abundance. J. Mamm., 36: 564-566.

Pearson, P. G. 1959. Small mammals and old field succession on the Piedmont of New Jersey. Ecology, 40: 249-255.

Zimmerman, E. G. 1965. A comparison of habitat and food of two species of Microtus. J. Mamm., 46: 605-612.

Manuscript received Sept. 30, 1970

ASSOCIATION OF PRODIGIOSIN WITH OUTER CELL WALL COMPONENTS

JOSEPH C. TSANG AND DENNIS M. KALLVY Department of Chemistry, Illinois State University, Normal, Illinois 61761

Abstract. — Prodigiosin was extracted along with outer membrane glycoprotein by sodium dodecyl sulfate and sodium de- oxycholate from isolated cell envelopes of Serratia marcescens 08. Part of the pig¬ ment was separated from the glycoprotein by organic solvent extraction and by Seph- adex G-200 column chromatography.

In Serratia marcescens there is a tripyrrole pigment, prodigiosin, which absorbs at 535-540 mu (red) in acid medium but becomes orange in color (470 mu) in alkaline medi¬ um. Prodigiosin has been studied by many workers because of its suspect¬ ed biological properties. Castro (Castro, et al., 1967) noted that prodigiosin was active against vari¬ ous pathogenic bacteria and fungi, while Allen (Allen, 1967) found that it acted as an auto-oxidizable elec¬ tron acceptor and suggested its pos¬ sible role in cellular respiration. Al¬ though the biosynthesis of the pig¬ ment has been worked out to some extent (Morrison, 1966), the exact location of biosynthesis and its site of association with cell particles re¬ mains unclear. Castro (Castro, et al., 1959), speculated that prodigio¬ sin was present in the cell as the salt of a fatty acid which associated close¬ ly with the lipid of cell membrane. However, the observation that the concentration of prodigiosin paral¬ leled that of N-acetylhexosamine in the isolated cell envelope indicated that the pigment may be associated

with the cell wall (Williams and Purkayastha, 1960). However, it is not certain whether the pigment is located in the outer or inner mem¬ brane of the cell envelope (DePetris, 1967). An extracellular glycopro¬ tein containing prodigiosin has also been isolated and characterized (Yo- shida, 1967). More recently, prodi¬ giosin was found covalently-bound to a glycoprotein of high molecular weight (Cruz-Camrillo and Sanchez- Zuniga, 1968). This report repre¬ sents the studv of the association of prodigiosin with the outer soft layer of the cell envelope.

Material and Methods

The cell walls were isolated from Serratia marcescens 08 (grown on an enriched medium) according to the procedure of Williams (Williams and Purkayastha, 1960). The cell wall preparation (fr. CW08) was extracted with dissociating reagents such as sodium dodecyl sulfate (SDS), sodium deoxycholate (SC), and guanidine hydrochloride (GH). One hundred milligrams of fr. CW08 was stirred in 50 ml of 0.5% SDS (pH 7.5) for one hour at 50°. After centrifugation at 10,000 r.p.m. for 20 minutes, the supernate and sedi¬ ment fractions were separated and SDS was removed by dialysis. After lyophilization, the supernate fraction

[22]

Tsang & Kallvy — Localization of Prodigiosin

23

(fr. SDS-S) and the sediment frac¬ tion (fr. SDS-P) were recovered. Similarly, extraction with 1% sodi¬ um deoxycholate yielded fractions SC-S and SC-P. The condition for guanidine hydrochloride extraction varied slightly. Three molar guani¬ dine hydrochloride solution was used at room temperature and fractions GH-S (supernate) and GH-P (sedi¬ ment) were recovered. Protein (Lowry, et al., 1951), glucosamine (Rondle and Morgan, 1955), hexoses (Koehler, 1952), and uronic acids (Bitter and Muir, 1962) were deter¬ mined. Total lipids were extracted with chloroform ; methanol (2 :1 v/v) in a soxhlet apparatus and deter¬ mined gravimetrically. Other or¬ ganic solvents extractions were per¬ formed in a similar manner. Pres¬ ence of prodigiosin was monitored with a Beckman DK-2A UV-Visible spectrophotometer after fractions SDS-S, SC-S, and GH-S were ex¬

tracted with ethanol :0.1 N HC1 (9 :1 v/v) mixture. Double diffusion tech¬ nique in agar gel (Ouchterlony, 1962) was used to study the im¬ munological activities of the various fractions against anti-whole cell serum. Endotoxin was isolated ac¬ cording to the modified method of Boivin (Tsang and Rilett, 1970).

Column chromatography of frac¬ tion SDS-S was performed in Sepha- dex G-200 which was equilibrated with 0.5% SDS. Thirty milligrams of the sample were dissolved in 3 ml of 0.005 M sodium phosphate buf¬ fer, pH 7.4 in 0.2% SDS. The col¬ umn was eluted by the same buffer and monitored by protein determina¬ tion.

Results and Discussion

The yields and the results of the chemical analyses are presented in Table 1.

Table 1. — Chemical Composition of Extracts from Isolated Cell Envelope from Serratia marcescens 08.

Fractions

Yield Per Cent

Protein Per Cent

Total Lipid Per Cent

Total

Carbohydrate Per Cent

Hexoses Per Cent

Hexosamine Per Cent

Uronic

Acid

Per Cent

SDS-S .

85

58.0

17.5

9.4

4.6

2.6

2.2

SC-S .

65

26.3

25.0

10.3

4.3

3.4

2.6

GH-S .

16

65.0

N.D.

8.2

3.0

3.8

1.4

It appears that the extracts were all glycoprotein in nature. Al¬ though fraction GH-S has the high¬ est content of protein (65%) and fraction SC-S has the highest total carbohydrate content (10.3%), the extracting effectiveness of sodium dodecyl sulfate was far superior to

the other two reagents. Guanidine hydrochloride proved to be a rela¬ tively poor extractant for the surface material (16%), and it also failed to remove any of the prodigiosin. On the other hand, both sodium dodecyl sulfate and sodium deoxycholate completely removed the pigment

24

Transactions Illinois Academy of Science

from the isolated cell envelope (Ta¬ ble 2). Despite the fact that pre¬ vious evidence indicated that hexo- samine and prodigiosin were present in the same cellular structure, such as the cell envelope (Williams and Purkayastha, 1960), our results of analysis with the extracts (fractions SDS-S, SC-S, and GH-S) do not show such parallel correlationship of content of hexosamine with the pres¬ ence of pigment (Table 2). Both fractions SDS-S and SC-S were pig¬ mented, but their hexosamine content was lower than their corresponding colorless sediment fractions (fr. SDS-P and fr. SC-P). This observa¬ tion suggested that prodigiosin was associated with certain components which could be selectively removed from the isolated cell envelope by mild dissociating reagents. These components may possibly be the gly¬ coproteins and/or other conjugated macromolecules, such as lipoglyco- protein complexes which are present on the outer soft layer of the wall. Indeed, glycoproteins have been re¬ moved from the outer cell membrane by sodium dodecyl sulfate (Wein- baum and Markman, 1966). It is surprising, however, that guanidine hydrochloride failed to perform the same function as the other two rea¬ gents since it has also been used for

removing outer surface components from walls of other Gram-negative bacteria (Kushner, 1969).

In order to demonstrate the im¬ munochemical similarities of the ex¬ tracted fractions, namely fractions SDS-S, SC-S, and GH-S and lipo- polysaccharide-protein complex (en¬ dotoxin) (Tsang and Rilett, 1970) from S. marcescens 08, anti-whole cell serum was allowed to react with these fractions by the double diffus¬ ion technique. All fractions tested with the exception of fr. GH-S gave one precipitate line which cross-re¬ acted with each other. It is interest¬ ing to note that antigenicity paral¬ lels with the presence of prodigiosin (Table 2). It is possible that guani¬ dine hydrochloride extracted the non-antigenic cellular glycoprotein rather than the outer-membrane gly¬ coprotein, while the other reagents removed glycoproteins which have at least one common antigenic compon¬ ent with endotoxin.

In order to study the possible link¬ age between prodigiosin and the cell envelope glycoproteins, fractions CW08, SDS-S, and SC-S were ex¬ tracted with acetone and ethanol, as well as chloroform-methanol (2:1 v/v). After organic solvent extrac¬ tions, the residues were colorless, while the extracts gave the charac-

Table 2. — Correlationship between Pigmentation, Hexosamine Content and Immunologica 1 Activities of Extracts and Residues.

Fractions

Pigmentation

Hexosamine Per Cent

Immunological

Activities

SDS-S .

+

2.6

Positive, 1 line N.D.*

SDS-P .

5.6

SC-S .

+

3.4

Positive, 1 line N.D.

SC-P .

4.6

GH-S .

_

3.8

Negative

N.D.

GH-P .

+

+

5.3

Endotoxin .

12.2

Positive, 1 line

* N.D. = not done

Tsang & Kallvij — Localization of Prodigiosin

25

teristic spectra of prodigiosin. Cruz- Camarillo claimed that prodigiosin was conjugated with a glycoprotein to form a soluble complex which could be broken down with ethanol (Cruz-Camarillo and Sanchez-Zuni- ga, 1968) into a colorless component and a colored compound of low molecular weight. Our observation does not agree entirely with the im¬ plication of a possible existence of covalent linkage only between the pigment and the glycoproteins. It is likely that at least part of the pig¬ ment was associated with the macro¬ molecule by hydrophobic bonds and/ or salt bridges.

In the column fractionation exper¬ iment we obtained a single peak which gave positive protein reaction. Figure 1 shows the elution pattern. After dialysis and lyophilization, a colorless amorphous material was re¬ covered in 90% yield. The pigment remained bound to the column, and attempts to elute it failed. A similar

Figure 1. Sephadex G-200 Column Chromatography of Fraction SDS-S. Vol¬ ume collected 3 ml per tube. Column was monitored by protein determination on 0.2 ml aliquots.

result was obtained when the column was equilibrated with 1% sodium deoxycholate and eluted with 0.2% sodium deoxycholate in the same buf¬ fer. The results of these experiments strengthen our belief that not all of the prodigiosin is bound to the gly¬ coprotein.

Thus, it appears that prodigiosin is associated with the outer cell wall component which is glycoprotein in nature. This glycoprotein shares at least one antigenic component with the lipopolysaccharide-protein (en¬ dotoxin) complex. Whether the pig¬ ment is synthesized by the mem¬ brane enzymes or synthesized in the cytoplasm and transported to the outer membrane of the cell wall re¬ mains to be elucidated.

Acknowledgment

The authors thank Dr. P. Alaupovic, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, for the anti¬ whole cell serum. Supported in part by grants from Research Corporation, Chicago, Illinois, and Illinois State University Re¬ search Committee.

Literature Cited

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Bitter, T. and H. H. Muir. 1962. A Modified Uronic Acid Carbazole Reac¬ tion. Anal. Biochem. 4:330.

Castro, A. J., A. L. Coruin, F. J. Wax- ham, and A. L. Beiby. 1959. Products from Serratia marcescens. J. Organic Chem. 24:455-459.

Castro, A. J., G. R. Furcolow, G. R. Gale, G. E. Means, and G. Tertzakian. 1967. Antimicrobial Properties of Pyr¬ role Derivatives. J. Med. Chem. 10:29- 32.

Cruz-Camarillo, R. and A. A. Sanchez- Zuniga. 1968. Complex Protein-prodi- giosin in Serratia marcescens. Nature 218: 567-568.

De Petris, S. 1967. Ultrastructure of the Cell Wall of Escherichia coli and the Chemical Nature of Its Constituent Lay¬ ers. J. Ultrastruct. Res. 19:45-83.

26

Transactions Illinois Academy of Science

Koehler, L. H. 1952. Differentiation of Carbohydrates by Anthrone Reaction of Rate and Color Instensity. Anal. Chem. 24:1576.

Kushner, D. J. 1969. Self-assembly of Biological Structures. Bacteriol. Review 33:302-345.

Lowry, O. H., N. J. Rosenbrough, A. L. Farr, and R. J. Randall. 1951. Pro¬ tein Measurement with the Folin Re¬ agent. J. Biol. Chem. 193:265-275.

Morrison, D. A. 1966. Prodigiosin Syn¬ thesis in Mutants of Serratia marcens- cens. J. Bacteriol. 91:1599-1603.

Ouchterlony, O. 1962. Diffusion-In- Gel Methods for Immunological Analysis, II in Progress in Allergy. S. Kanger (Ed.) Vol. VI. New York.

Rondle, G. J. M. and W. T. J. Morgan. 1955. The Determination of Glucosa¬ mine and Galactosamine. Biochem. J. 61:586.

Tsang, J. and J. Rilett. 1970. Isolation

and Fractionation of Lipopolysacchar- ides from Serratia marcescens Bizio. Trans. Ill. State Acad. Sci. Vol. 63: 324-328.

Weinbaum, G. and R. Markman. 1966. A Rapid Technique for Distinguishing Enzymatically Active Proteins in the Cell “Envelope” of Escherichia coli. Biochim Biophys. Acta. 124:207.

Williams, R. P., A. Green, and D. A. Ra- poport. 1956. Studies on Pigments from Serratia marcescens. T. Bacteriol. 71:115.

Williams, R. P. and M. Purkayastha. 1960. Association of Pigment with the Cell Envelope of Serratia marcescens. Nature 187:349-350.

Yoshida, S. 1962. Study of a Soluble Complex of Prodigiosin Produced by a Strain of Serratia marcescens. Can. J. Biochem, Physiol. 40:1019.

Manuscript received September 2, 1970

WOODY VEGETATION OF HART MEMORIAL WOODS, CHAMPAIGN COUNTY, ILLINOIS

T. W. ROOT, J. W. GEIS, and W. R. BOGGESS Department of Biology, Blackhawk College, Moline, Illinois; Department of Forest Botany and Pathology, New York State College of Forestry, Syracuse University, Syracruse, New York; and Department of Forestry, University of Illinois, Urbana

Abstract. — The Hart Memorial Woods is one of the more xerophytic examples of upland streamside forests in the Prairie Peninsula of east-central Illinois. Upland soils are well developed and support a mixed stand of white oak (Quercus alba L.), black oak (Q. velutina Lam.), and red oak ( Q . rubra L.). Composition and ecological trends of three physiographic units (bottom¬ land, upland, and mixed) are discussed. Hickories appear to be becoming more important stand components, based on num¬ bers of seedlings and saplings present. As is characteristic of other woodlands studied, the oaks are not reproducing well. Elm mortality has been extremely heavy in the bottomland and mixed physiographic units.

The Hart Memorial Woods, lo¬ cated along the east bank of the Sangamon River near Mahomet, Illi¬ nois, is one of the more xerophytic examples of upland streamside for¬ ests in the Prairie Peninsula of east- central Illinois. The woodland is a remnant of a much larger timbered area that was about three miles wide and extended northward for six miles along the Sangamon from Mahomet (Spaeth, 1963) . Hart Woods was ac¬ quired by the University of Illinois in 1965 for its system of natural areas. These areas now complete a sequence from the wettest to the dri¬ est upland forest sites in the area (Boggess, 1964; Boggess and Bailey, 1964 ; Boggess and Geis, 1966 and 1967). Among the natural areas, Hart Woods is unique in that it pro¬ vides a transect from moist bottom¬ lands, formerly dominated by elm, to an upland series occupied largely

by red oak, white oak, and black oak as the sites become progressively drier. The woody vegetation of this woodland and its general ecological status are discussed in this paper.

Description of Area

Hart Memorial Woods occupies the N1/^ and E 16 acres of the S1/^, NEi/4, SW14, S36, T21N, R7E, 3rd P.M. (40° 14' N. Lat.; 88° 21' W. Long.), Champaign County, Illinois. The area lies between elevations of approximately 672 and 703 feet above sea level and includes a level bottomland and slopes up to 30 per¬ cent. Two small streams pass through the area and empty into the Sangamon River. One is inter¬ mittent, while the other has some flow except during prolonged dry weather.

Soils

Some of the most highly developed soils in the prairie-forest border of central Illinois occur on the uplands of Hart Woods. They are recognized as Gray -Brown Podzolic soils in the classification of Thorp and Smith (1949) and as Hapludalfs in the cur¬ rent system detailed in the 7th Ap¬ proximation (Soil Survey Staff, 1960). The Birkbeck and Camden series are the most prevalent upland soils in the woodland, and both de¬ veloped in loess under the influence of forest vegetation. Birkbeck is moderately well-drained and devel-

[27]

28

Transactions Illinois Academy of Science

oped in 36 to 60 inches of loess over glacial till. Camden, a shallower soil, developed in 15 to 36 inches of loess over glacial ontwash and is well-drained. Horizons in both series are distinct and easily differentiated. The dark gray to brown surface lay¬ ers are silt loams and grade into yel¬ lowish brown silty clay loam subsoils. Profiles are acid throughout.

One representative profile each of Birkbeck and Camden was excavated, described, and sampled. Data on se¬ lected physical and chemical charac¬ teristics are shown in Table 1. These data clearly indicate that the better upland sites are associated with

the Birkbeck soils and that they sup¬ port a more mesic vegetation than does Camden. Cation exchange ca¬ pacity and percent base saturation is greater throughout the Birkbeck profile indicating its superior nu¬ trient status as compared with Cam¬ den. Moisture relations are more favorable in Birkbeck due to gener¬ ally higher amounts of fine-textured materials at all depths, greater amounts of organic carbon in the A horizon, and character of materials underlying the solum. Within the woodland, Camden soils are associ¬ ated with stands that run heavily to black oak.

Table 1. — Selected physical and chemical characteristics of Camden silt loam and Birk¬ beck silt loam.

Texture, %

Cation

Exch.

Base

Horizon

Depth,

Inches

> 2 mm.

Sand

Silt

Clay

pH

Organic

Carbon,

%

Cap., me/ 100 gm.

Satura¬

tion,

%

Camden silt loam

A1 .

0- 3

0.13

8.8

71.4

19.8

4.86

2.31

9.71

36.2

A21 .

3- 6

0.05

9.2

69.0

21.8

4.71

0.91

8.80

39.8

A22 .

6-12

0.10

9.0

66.2

24.8

4.90

0.46

9.30

58.1

B1 .

12-18

0.11

7.0

62.0

31.0

4.97

0.25

13.47

72.5

B21 .

18-24

0.03

6.6

56.6

36.8

4.99

0.25

18.53

75.2

B22 .

24-31

0.00

11.2

53.0

35.8

4.88

0.14

18.56

75.2

II B23 .

31-42

0.00

21.1

50.1

28.8

4.68

0.12

17.38

71.2

II B31 .

42-63

0.20

22.0

53.0

25.0

4.70

0.10

13.01

66.4

II Cl .

63-75 +

0.28

68.2

18.3

13.5

4.64

0.03

8.86

68.4

Birkbeck silt loam

A1 .

0- 4

0.45

5.8

67.8

26.4

4.98

3.18

26.55

91.7

A21 .

4- 8

0.61

6.0

66.0

28.0

4.90

1.24

15.54

85.8

A22 .

8-12

0.60

4.1

65.4

30.5

4.89

0.49

14.68

85.0

B1 .

12-18

0.16

3.6

64.0

32.4

5.07

0.27

21.70

84.8

B21 .

18-26

0.04

2.5

59.5

38.0

5.10

0.14

30.10

86.0

B22 .

26-36

0.06

1.9

57.0

41.1

4.90

0.14

22.89

81.6

B31 .

36-52

0.07

8.0

60.4

31.6

4.87

0.08

21.74

84.8

II Cl .

52-59

2.29

13.9

60.7

25.5

4.80

0.10

20.96

92.8

II C2 .

59-68

9.86

24.5

48.1

27.4

4.78

0.07

13.47

97.0

II C3 .

68-95 +

12.00

31.2

46.2

22.6

4.59

0.06

28.29

100.0

Root et al — Hart Woods

29

Bottomland soils include both medium -textured (loam and silt loam) and moderately fine-textured (silty clay loam) alluvial soils. Medium-textured soils are represent¬ ed by Otter silt loam, and the fine- textured by Sawmill silty clay loam. Both of these soils are imperfectly drained. New soil material is added to the bottomland almost every year, as spring flods deposit soil from the intensively cultivated uplands in the surrounding area. Because of this continuing deposition, soil horizons are poorly defined. Nutrient and moisture conditions are quite favor¬ able for tree growth.

Methods

Prior to inventory in 1965, the woodland was permanently divided into 50-meter square blocks. Inven¬ tory data were kept separately by quarter-blocks, designated by divid¬ ing each block along the diagonals. Diameters at 4% feet above the ground (DBH) of all woody vegeta¬ tion 2.6 inches and above, were meas¬ ured and tallied by species. Dead¬ standing and dead-down trees were also measured and identified where possible. Four sets of nested circular plots (1/100 and 1/1,000 acre) were randomly located in each sample block. Small saplings (1 and 2 inch¬ es DBH) were measured on the larg¬ er plots, and seedlings on the smaller plots. Seedlings were tallied by spe¬ cies and height class, those less than one foot tall, and those greater than one foot in height but less than 0.6 inch DBH.

Stand data were developed by summarizing information for quar¬ ter-blocks that fell completely in the bottomland, upland, or partly in both. The latter is designated as a “mixed” topographic unit and in¬ cludes areas that are transitional be¬

tween the two main topographic un¬ its. Although the “mixed” category is highly artificial, it allows a more concise interpretation of the bottom¬ land and upland data.

Results

A total of 34 woody species were tallied. These are shown, along with their density and frequency by size class, in Table 2. The number of trees and basal area per acre, rela¬ tive density, relative dominance, and Importance V alue for the 10 leading species in each topographic unit are shown in Table 3. As used here, Im¬ portance Value (IV) is that defined by McIntosh (1957) and is the sum of the relative dominance and rela¬ tive density (Boggess and Geis, 1967). The leading dominant (spe¬ cies with the highest IV) is shown for each quarter-block in Fig. 1, and a breakdown for these same species into broad diameter classes is shown in Table 4.

wo

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Figure 1. Diagram of woodland showing species with highest Importance Value by quarter-blocks.

Table 2. — Checklist of woody taxa identified and number per acre and frequency (percent) of seedlings and sapling by species and physiographic

30

Transactions Illinois Academy of Science

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Totals .

32

Transactions Illinois Academy of Science

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34

Transactions Illinois Academy of Science

Upland Physiographic Unit

White oak and black oak rank first and second in importance. Red oak is in fourth position, slightly behind red elm. As a group, these three oak species comprise 70 percent of the total number of trees and 95 per¬ cent of stand basal area. Red oak and black oak have the largest dia¬ meters of all trees in the woodland, averaging 17.9 and 17.5 inches, re¬ spectively. Black oak is concentrat¬ ed on the steeper slopes in the north¬ eastern corner of the woods, where it exceeds white oak in importance. Seedling counts are 290 per acre for white oak and 507 per acre for black oak. Black oak was not represented in the 1- and 2-inch diameter classes, and there were only 5 white oaks per acre present in this size class.

Slippery elm and American elm ranked third and eighth in IV. Be¬ cause of the high mortality of larger trees from phloem necrosis and Dutch elm disease, these two species have the smallest diameters of any leading dominant. Red elm com¬ prises 30 percent of the seedlings, 35 percent of the small saplings, and almost one-tliird of the trees in the 3- to 6-inch diameter class. Its fre¬ quency of 77.6 percent for seedlings and 64.5 percent for small saplings exceeded that of any other species.

Black cherry, although confined largely to the smallest diameter classes, ranked fifth in IV and had the second highest number (2,956) of seedlings per acre. Unlike red elm, black cherry does not continue its high density into the 3- to 6 -inch diameter class. Mortality of seed¬ lings appears to be particularly hea¬ vy from 3 to 5 years after establish¬ ment.

In some parts of the upland, sixth- ranked sassafras forms a dense un¬ derstory as evidence by the fact it

accounts for almost one-third of the 1- and 2-inch trees. The 862 sassa¬ fras seedlings per acre, with a fre¬ quency of 30 percent, indicates that this species is reproducing quite well.

Three hickories — mockernut, shag- bark, and bitternut — rank seventh, ninth, and tenth in IV, respectively. Collectively, there are 950 hickory seedlings per acre and 79 trees in the 1- and 2-inch diameter classes.

Other species in the upland for¬ est, along with their IV ’s, are black walnut, 1.01; redbud, 0.54; shingle oak, 9.52; basswood, 0.32; red mul¬ berry, 0.31 ; honey locust, 0.25 ; iron- wood, 0.13; hawthorn, 0.06; green ash, 0.06 ; and hackberry, 0.04.

Mortality on the upland amounted to 6.9 square feet of basal area per acre. This included an average of about 7 elms and 8 white oaks per acre, with an occasional tree of other species.

Bottomland Physiographic Unit

Stocking in the bottomland is only one-half that of the upland, both from the standpoint of tree number and basal area. This is due to the near complete mortality of elm that once composed almost half of the bottomland stand. Dead-standing and dead-down elm comprise 65 trees and 48 square feet of basal area per acre. Total mortality (all species) in the bottomland was 70 trees and 56 square feet of basal area per acre.

Silver maple is the most important species and includes 23 percent of the trees and 40 percent of the basal area. Green ash is second in impor¬ tance, followed by American elm in third place. The elms, including red elm, which is sixth in IV, are less than 6 inches in DBH. In contrast the largest tree in the entire wood¬ land is a 49-inch green ash. Other species included in the 10 leading

Root et al — Hart Woods

35

dominants with their IV ’s are hack- berry, 14.1 ; black walnut, 11.7 ; hon¬ ey locust, 7.0; bur oak, 6.8; shingle oak, 5.6 ; and hawthorn, 5.5. Collec¬ tively the IV of the “others” cate¬ gory is almost as great as that of silver maple, stressing the relatively high importance of minor species in the bottomland as compared with the upland.

Regeneration of woody species in the bottomland is sparse as indicated by the tally of 83 seedlings and 98 small saplings per acre. The elms were reproducing better than any other tree species.

Mixed Physiographic Unit

Since this unit is transitional, it contains species characteristic of both the bottomland and upland areas. It also contains the highest quality sites in the woodland. While there are more trees per acre pres¬ ent in the upland, 146 vs. 128, the basal area of 97 square feet per acre is about 17 sq. ft. less than that of the upland. Again, elm mortality has been an important factor, amounting to 39 trees and 35 sq. ft. of basal area per acre.

White oak, with an IV of 40.1, is the leading dominant, followed close¬ ly by red oak (IV, 31.8). However, the 30 white oaks per acre have an average diameter of 10.9 inches com¬ pared with 21.5 inches for the 10 red oaks present. Black oak is third in importance and is intermediate in size (average DBH, 16.4 in.) be¬ tween the white and red oak. Collec¬ tively, these three oak species com¬ prise 35 and 63 percent, respective¬ ly, of number of trees and basal area. Two additional oak species, shingle and bur, rank sixth and tenth, re¬ spectively, in importance but consti¬ tute a minor part of the stand. Re¬ generation of the oaks presents about

the same picture as in the upland.

Other than oaks, the elms are the second most important species, with red elm ranking fourth and Ameri¬ can elm fifth in importance. Their IV is based on tree number rather than size, with most of the individ¬ uals falling in the 3- to 6-inch dia¬ meter class. The former position of American elm in the stand is il¬ lustrated by the fact that dead in¬ dividuals have an average diameter of 12.8, compared with 4.1 for those still living. However, larger individ¬ uals of red elm persist in the stand, especially in this transitional physio¬ graphic position. Also, red elm re¬ generation is greater than that of any other species, comprising 35 per¬ cent of the 5,178 seedlings and 50 percent of the 283 small saplings (1- and 2-inch DBH) per acre in the stand. Amrican elm is poorly repre¬ sented in both of these size classes, reflecting the lack of seed source.

Discussion

The positioning of Hart Woods upland at the xeric end of a mois¬ ture sequence for upland forests in east-central Illinois is justified by two factors: (1) the complete ab¬ sence of sugar maple ( Acer sacchar- um Marsh) in the stand; and (2) the greater importance of white oak and black oak, particularly the lat¬ ter, compared with other woodlands studied. On more mesic sites in these woodlands, sugar maple is an extremely aggressive species and ap¬ pears to be continually increasing in importance. Although sugar maple ranked only ninth in IV in the streamside forest at Allerton Park, it is an important stand component with a relatively large number of individuals in the 3- to 6-inch dia¬ meter class (Boggess and Geis, 1967).

White oak will probably increase

36

Transactions Illinois Academy of Science

in importance as the larger black oaks die. There are, as replacements, 31 white oaks compared with only 3 black oaks per acre less than 13 inches DBH. Mortality data sug¬ gests this trend, as the basal area of dead black oaks in the largest dia¬ meter class (13 to 24 inches) is four times that of white oak. Although mortality of small white oaks (3- to 6-inch DBH) is relatively high, there remain more live than dead trees. The reverse is true for black oak. Then, too, back oak does not gener¬ ally reproduce well in the absence of some disturbance factor (U. S. Forest Service, 1965). Any predic¬ tion of future composition must be tempered by knowledge of the recent devastation of the elm population, plus the fact that oak wilt is a threat in sections of Illinois.

Although hickories as a group ranked relatively low among the leading dominants, they were third in the number of seedlings and small saplings. On this basis, hickories are likely to increase in importance.

The future of red elm is an intrig¬ uing question, as seedlings and sap¬ lings are quite dense in parts of the upland. However, many individuals were produced from interconnecting rhizones rather than seed. There is a sharp drop in the density of red elm above 6 inches in diameter which suggests that the relatively large number of indiivduals less than this diameter may be of relatively recent origin. Competition from red elm in these smaller diameters has undoubtedly affected oak regenera¬ tion, particularly the growth of seed¬ lings into saplings and so on. Any suggestion that red elm will form an important component of the future overstory must consider the suscepti¬ bility of this species to Dutch elm disease and phloem necrosis, even

though it is less than that of Amer¬ ican elm.

The presence of large numbers of sassafras seedlings and small sap¬ lings in the upland forest is un¬ usual for this area. This species did not occur in Trelease Woods, Brown¬ field Woods, or the Funk Forest Natural Area. Although sassafras did occur on well-developed forest soils, Allerton Park numbers were quite limited compared with the Hart woodland. Sassafras along with per¬ simmon ( Diospyros virginiana D.) are usually the first woody plants to appear in fields abandoned from cultivation in southern Illinois. While sassafras persists for many years, it appears to drop out of the successional picture in mature oak- hickory forests of the area (Bazzaz, 1968). In sharp contrast, Eisen- hauer (1967) found that seedlings of sassafras ranked second in im¬ portance in forests growing on rela¬ tively level claypan soils and on slopes in south-central Illinois. In these same stands, it ranked eleventh in IV for trees 3 inches DBH and above. Sassafras is found in Sar¬ gents and Baber woods, located on the Shelbyville Moraine approxi¬ mately 50 miles southeast of Hart Woods, occurring as seedlings, small saplings, and small trees exceeding 3 inches DBH. (Ebinger, 1968; Mc¬ Clain and Ebinger, 1967). It ranks eleventh in importance in Baber Woods but is not included among the 10 most important species in Sargents Woods.

Hart Woods is at the northern range of sassafras in east-central Il¬ linois, although farther to the east it extends into central Michigan. Har¬ mon (1968) reported abundant sas¬ safras in the dune forests of south¬ western Michigan, as did Olson (1958) from the Indiana dunes.

37

Boot et al — Hart Woods

Thus the distribution of sassafras (U. S. Forest Service, 1965) shows a distinct adjustment to the geogra¬ phic occurrence of the Prairie Penin¬ sula. Kucera and McDermott (1954) list sassafras as one of the species common to Missouri forests that drops out in the northern portions of the Prairie Peninsula. The abun¬ dance of sassafras in Hart Woods may well be related to the advanced stage of soil development that has resulted in more pronounced base re¬ moval and lower pH than is found for soils of other woodlands in the area.

The future composition of the bot¬ tomland forest is uncertain. Spring flooding, siltation, heavy soils, and a dense herbaceous cover will all affect the establishment of tree seedlings. Silver maple is now the leading dom¬ inant, but whether it will fill the niche created by loss of the elms is open to question.

Literature Cited

Bazzaz, Fakhri A. 1968. Succession on abandoned fields in the Shawnee Hills, Southern Illinois. Ecology 49: 924-936.

Boggess, W. R. 1964. Trelease Woods, Champaign County, Illinois: woody

vegetation and stand composition. Trans. Ill. State Acad. Sci. 57: 261-271.

- , and L. W. Bailey. 1964.

Brownfield Woods, Illinois: woody vege¬ tation and changes since 1925 . American Midland Naturalist 71: 392-401.

- , and J. W. Geis. 1967. Com¬ position of an upland, streamside forest in Piatt County, Illinois. American Mid¬ land Naturalist 78: 89-97.

- , and - . 1966. The

Funk Forest Natural Area, McLean

County, Illinois: woody vegetation and ecological trends. Trans. Ill. State Acad. Sci. 59: 123-133.

Ebinger, J. E. 1968. Woody vegetation survey of Sargeants Woods, Coles County, Illinois. Trans. Ill. State Acad. Sci. 61: 16-25.

Eisenhauer, Leon D. 1967. Relation¬ ships between oak-hickory forest types and soils in central Illinois. Ph.D. Thesis, University of Illinois at Urbana-Cham- paign. 77 pp.

Harmon, Jay R. 1968. Environment and forest gradients on the southeast shore of Lake Michigan: a study in plant geography. Ph.D. Thesis, University of Illinois at Urbana-Champaign. 88 pp.

Kucera, C. L., and R. E. McDermott. 1954. Sugar maple-basswood studies in the forest-prairie transition of central Missouri. American Midland Naturalist 54:495-503.

McIntosh, R. P. 1957. The York Woods: a case history of forest succession in southern Wisconsin. Ecology 38: 29-37.

McClain, W. E., and J. E. Ebinger. 1967. Woody vegetation of Baber’s Woods, Edgar County, Illinois. American Mid¬ land Naturalist 79: 419-428.

Oison, Jerry S. 1958. Rates of suc¬ cession and soil changes on southern Lake Michigan sand dunes. Bot. Gaz. 119: 125-170.

Soil Survey Staff (1960). Soil classifica¬ tion, a comprehensive system, 7 th Ap¬ proximation. Soil Conserv. Serv., USDA. 265 pp.

Spaeth, J. N. “Forests,” Natural Re¬ sources of Champaign County. Ed., D. F. Hansen. Champaign County Conser¬ vation Education Council, Urbana, Ill. pp. 5-8.

Thorp, J., and Guy D. Smith. 1949. Higher categories of soil classification: order, suborder, and great soil groups. Soil Sci. 67: 117-126.

U. S. Forest Service. 1965. Silvics of forest trees of the United States. U.S. Department of Agriculture Handbook no. 271.

Manuscript received July 8 , 1970

GROWTH RELATIONSHIPS BETWEEN APHELENCHUS AVENAE AND TWO SPECIES OF NEMATOPHAGOUS FUNGI

H. L. MONOSON

Biology Department, Bradley University, Peoria, Illinois

Abstract. — Comparable populations of the mycophagous nematode, Aphelen- chus avenae Bastian, were cultured for seven days upon agar cultures with two species of nematophagous fungi. In the presence of nematodes the optimum growth temperature of the fungi was usually in¬ creased 5°C and fungus colony diameters were greater than controls. Low numbers of living nematodes were recovered from fungus-sown plates after seven days at 15°C and 20°C, but at 25°C and 30°C the numbers of nematodes recovered exceeded the original inoculum. Nematode numbers at the lower temperatures suggested that nematode trapping was higher than nema¬ tode reproduction, eggs laid by the nema¬ todes were incapable of hatching, or in¬ hibitory fungal metabolites are stable at lower temperatures and unstable at higher temperatures.

Experimental cultures of nema¬ tode-trapping fungi have included primarily either bacterium-feeding or plant-parasitic nematode species. Mycophagous nematodes have been used only recently in such cultures even though they are probably as widely distributed in nature as are the nematophagous fungi. Recent investigations (Cayrol, 1967; Cooke and Pramer, 1968 ; Feder, 1963 ; Hechler, 1963 ; Monoson, 1968a) have included a study of the relation¬ ships between nematophagous fungi and mycophagous nematodes.

The purpose of the present study was to determine what effects vary¬ ing numbers of nematodes have on fungus growth and what effects the fungi have on nematode populations when both are grown together.

Materials and Methods

Two species of nematophagous Moniliales were chosen. The fungi used in this study were two adhesive, network-forming species, Arthrobo- trys oligospora Fres. and A. musi- formis Drechs. Stock cultures of the fungi were maintained on a medium containing 20 g Difco corn-meal agar (CMA) in 1000 ml water at room temperature. The nematode Aphe- lenchus avenae Bastian was main¬ tained on Pyrenochaeta terrestris (Hansen) Gorenz, Walker, and Lar¬ son which grew on a medium con¬ taining 10 g Difco potato-dextrose agar (PDA) and 15 g agar in 1000 ml water.

Three media were used in this study: 2% Difco CMA, one-quar¬ ter-strength Difco PDA (Monoson, 1968a), and one-fifth-strength Y-8 agar (V-8) composed of 200 ml V-8 juice@ and 20 g agar in 1000 ml water. No attempt was made to ad¬ just the pH’s of the media.

Fungus inoculations were accord¬ ing to the technique described by Monoson (1968a). Each of the two fungi was cultured on the three agar media for a period of four days. Fungus plugs, 7 mm diam., were cut from these four-day-old cultures and placed in the center of 9 cm petri plates that contained the same agar medium. Nematodes were extracted from stock cultures according to the following technique (Monoson.

[38]

Mono son — Nematophagous Fungi

39

1968b) : Nematodes were separated from agar stock cultures by pouring 10-15 ml of sterile water over the surface of a plate. The nematodes readily moved into the surface water and were unable to re-enter the agar. A simple nematode extraction ap¬ paratus was formed by placing four layers of type 900-S-Kimwipe® tis¬ sues between two plastic funnel tops which rested in a Syracuse dish. The nematode extraction apparatus had been sterilized before hand by auto¬ claving for 15 minutes at 20 psi. Nematodes were extracted over a 1- hour-period and serially diluted to either 100 or 400 individuals per milliliter of water. Inoculation of either 100 or 400 nematodes into each experimental fungus culture was ac¬ complished by using a sterile meas¬ uring dispenser or a calibrated 1 ml duplicating pipette. Each culture was replicated three times and ap¬ propriate controls containing fungus alone were used.

Nematophagous fungus cultures containing A. avenae were main¬ tained for seven days at 15°, 20°, 25°, or 30° C. Observations and measurements of cultures began 24 hr after inoculation and continued at 24-hr intervals. The radial diameter of colonies was measured to the near¬ est 0.5 mm.

Upon completion of each test the agar contents of a petri dish were cut into approximately 0.5 cm squares and homoginized in a War¬ ing blendor for 8 sec. The homog¬ enate was poured into a nematode re¬ covery apparatus (Monoson, 1968b) and counts of the living nematodes were made after 24 hr.

Results

Average colony growth of the two fungi in radial diameter in the pres¬

ence and absence of A. avenae are shown in Figure 1. A. oligospora and A. musiformis had maximum growth at 25 °C in the presence of the nema¬ tode. A. musiformis grew more at 25° than at 20 °C on CM A. The growth of A. oligospora on PDA with A. avenae was as great at 20° as at 25° C. Slight growth differ¬ ences were noted for both fungi on CMA in the presence of the nema¬ tode at 20° and 25° C but were not considered significant.

Neither of the fungi repelled the nematode although in two instances less than 35% of the total number introduced was recovered from an

i ARTHROBOTRYS OLIGOSPORA e 50

15 20 25 30

TEMPERATURE C

15 20 25 30

TEMPERATURE C

Figure 1. Average colony growth of two species of nematophagous fungi on three agar media after seven days of growth with and without Aphelenchus avenae.

The bars are arranged in pairs. The solid black is the left-hand member of the pair, and gives the average colony growth in the presence of the nematode with an initial inoculum of 100 individuals. The right-hand member of the pair gives the average colony growth in the absence of the nematode, and is appropriately shaded to designate the medium, (see key).

40

Transactions Illinois Academy of Science

experimental culture (Table 1). All of the nematodes inoculated into a dish settled in the fungus plug after 12 hr. The nematodes moved freely in and around the inoculum plug prior to the 12 hr but were not ob¬ served to leave a colony once they began feeding on the hyphae. Fungal satellite colonies were never formed through the transportation of spores on the bodies of nematodes. At no time were immobile nematodes ob¬

served other than those which had been captured by a fungus.

Cultures were evaluated on the basis of nematodes recovered alive. Low numbers of nematodes were re¬ covered from all the media main¬ tained at 15° and 20° C, but at 25° and 30 °C numbers were much high¬ er than used as inoculum (Table 1). The number of live nematodes re¬ covered was divided by the total area of the colony, in square centimeters,

Table 1. — Total numbers of Aphelenchus avenae recovered per plate* after being cultured seven days with two nematophagous fungi on various agar media.

Temperature

No.

Nematodes

Added

No. Nematodes Recovered

Arthrobotrys Oligospora

Arthrobotrys Musiformis

CMA

PDA

V-8

CMA

PDA

V-8

15° C .

100

52

32

28

56

42

47

15° C .

400

56

124

44

74

111

41

20° C .

100

74

69

42

56

54

79

20° C .

400

61

295

56

219

181

32

25° C .

100

231

130

126

835

238

87

25° C .

400

1500

445

254

1600

406

139

30° C .

100

890

296

690

2600

793

834

30° C .

400

5000

919

661

3900

339

427

* Numbers recorded are averages of three replicates in each case.

to vield the number of nematodes present per square centimeter of fun¬ gus (Table 2).

Discussion

The addition of A. avenae to fun¬ gus cultures usually resulted in more fungus growth compared to cultures without the nematode. Cultures that contained 100 nematodes as inocu¬ lum most often produced the op¬ timum fungus growth in this study.

Monoson (1968a) reported that A. ave?iae was captured very effective¬ ly by these two fungi after four days

of maintenance at 15°, 20°, 25°, and 30° C on 2% CMA, one-quarter- strength PDA, and one- fifth - strength V-8. In the present study, the low numbers of living nematodes recovered at 15° and 20° C indicated that nematode trapping might have occurred after the seven days of these tests. The data also suggested that nematode trapping was higher than nematode reproduction or that eggs laid by the nematodes were incap¬ able of hatching.

Cooke and Framer (1968) report¬ ed that nematode-trapping fungi dis¬ played no predaceous activity until

Monoson — Nematophagous Fungi

41

Table 2. — Numbers of Aphelenchus avenae per cm2 recovered after seven days of growth with two nematophagous fungi on various agar media.*

Temperature

No.

Nematodes

Added

Arthrobotrys Oligospora

Arthrobotrys Musiformis

CMA

PDA

V-8

CMA

PDA

V-8

15° C .

100

1

1

2

3

2

6

15° C .

400

2

3

3

5

6

6

20° C .

100

1

1

2

1

4

6

20° C .

400

1

5

2

7

7

2

25° C .

100

4

2

3

17

7

3

25° C .

400

24

7

4

39

22

7

30° C .

100

60

6

48

2300

202

629

30° C .

400

602

328

52

9750

484

237

* Average of three replicates.

they had completely colonized the agar substrate. In addition, they stated that the retardation of preda¬ ceous activity was due to the con¬ centration of nematodes in a zone at or beyond the colony margin too juvenile to capture nematodes. It was of interest to note that neither of these situations were observed in the present study.

Temperature affected the amount of fungal growth in the same way both with and without nematodes (Figure 1). With the exception of A. oligospora on PDA fungus cul¬ tures that contained nematodes had an optimum temperature for growth five degrees above that of the con¬ trols.

Large amounts of fungus growth and high numbers of live nematodes were recovered at either 25° or 30° C on the three media. Released nema¬ tode metabolic products may have caused the stimulated growth of the fungi due to alteration of the agar media. Small shifts in the pH’s of tiie media were noted but not enough data were available to ascribe spe¬ cific effects to pH shifts.

Monoson (1968a) reported that 20- 50 nematodes were recovered from

CMA cultures of A. oligospora and A. nmsiformis after four days at 25 °C when the inoculum level was 100 nematodes. Almost identical numbers of live nematodes were re¬ covered under those same environ¬ mental conditions when the inoculum level was 400 individuals. In the present study, similar fungus cul¬ tures on CMA at 25 °C with an inocu¬ lum level of 100 nematodes contained 231, 835, and 373 nematodes after seven days. These numbers were con¬ sidered low when compared with those produced on P. terrestris (a non-predaceous fungus) after a sim¬ ilar maintenance period at 25 °C. The number of nematodes expected on P. terrestris cultures after seven days was either equalled or surpass¬ ed, for the most part, during a sim¬ ilar period of time on A. oligospora and A. musiformis, at the two higher temperatures.

The large nematode numbers could be interpreted as being due to : non¬ functioning of the trapping mechan¬ ism after a seven-day period at high temperatures — i.e., 25° and 30° C, (2) increase of nematode reproduc¬ tion over the rate of nematode trap¬ ping, or (3) decrease in generation

42

Transactions Illinois Academy of Science

time of the egg to the larval and adult stages because of cultural con¬ ditions. Microscopic observation of whether or not a trap still functioned after seven days was considered im¬ possible because of the amount of fungal growth.

Acknowledgments

Thanks are due to Dr. Helen C. Hechler for providing the nematode culture; Drs. R. C. Cooke, W. A. Feder, and A. C. Tar- jan for providing the fungus cultures.

Literature Cited

Cayrol, J. C. 1967. Etudes preliminaires cfes relations entre quelque champignons pathogenes des vegetaux et deux especes de nematodes mycophages: Ditylenchus myceliophagus j. B. Goodey, 1958 et Aphelenchoides composicola M. T.

Franklin, 1957. Ann. Ephiphyt. 18: 317- 329.

Cooke, R. C. and D. Pramer. 1968. In¬ teractions of Aphelenchus avenae and some nematode-trapping fungi in dual culture. Phytopathology 58: 659-661.

Feder, W. A. 1963. A comparison of nematode-capturing efficiencies of five Dactylella species at four temperatures. Mycopath. Mycol. Appl. 19: 99-104.

Hechler, Helen C. 1962. The develop¬ ment of Aphelenchus avenae Bastian, 1865 in fungus culture. Proc. Helminthol. Soc. Wash. 29: 162-167.

Monoson, H. L. 1968a. Trapping effec¬ tiveness of five species of nematophagous fungi cultured with mycophagous nema¬ todes. Mycologia 60: 788-801.

- . 1968b. Methods of cultiva¬ tion, isolation, and inoculation of myco¬ phagous nematodes into nematophagous fungus cultures. Trans. Ill. St. Acad. Sci. 61: 210-211.

Manuscript received January 13, 1970

TRANSVERSE AND CRESCENT CRACKS IN SOYBEAN COTYLEDONS ASSOCIATED WITH IMBIBITION

KUO-CHUN LIU AND A. J. PAPPELIS

Department of Botany, Southern Illinois University, Carbondale, 62901

Abstract. — Transverse and crescent cracks in cotyledons and cotyledon fracture were induced in soybean seeds by placing them in water (room temperature) prior to planting. These injuries resulted in reduced rates of seedling growth or non¬ emergence, depending on location and severity of the cotyledon damage. The cracks also were induced by allowing seeds to swell for 12 hours on wet filter paper at 10 degree intervals from 10 to 60°C. The highest percentage of cotyledon crack¬ ing (100%) was observed at the lowest temperature and the lowest percentage (19%) at the highest temperature. These cotyledon cracks were apparently caused by uneven swelling during imbibition.

In a study of cell death in pith tissue of soybean ( Glycine max L.) seedlings, Liu (1966) often observed transverse and crescent cracks in the upper surface of cotyledons of young seedlings in all 24 varieties studied. By anticipating a small percentage of damaged seedlings in each exper¬ iment and overplanting to permit se¬ lection of uniform seedlings having no obvious cotyledon damage, Liu was able to complete the study of parenchyma cell death in pith tissue of normal seedlings. This type of cell death also occurred in seedlings with injured cotyledons. For future studies of the cell death process in soybeans, more uniform stands are desired. Thus, an explanation of the cotyledon injury was required.

We thought that cotyledon injury could be due to injuries to the seed during harvesting (Bainer and

Borthwick, 1934). More recent stud¬ ies suggested that soybean cotyledons could be injured by preharvest mois¬ ture conditions and during storage (Metzer, 1967 ; Tachibana, et al., 1968). Earlier literature indicated that cracked cotyledons could result due to rough harvesting and clean¬ ing treatments (Humphrey, 1958; Moore, 1957). Cracks in cotyledons reduced field stands due to weak¬ ened plants even when the seeds were treated with fungicides and planted under favorable field conditions. In¬ ternal injuries in those reports in¬ cluded bruised and fractured seed leaves, roots, and plumules with com¬ plete or partial fractures noted in many plant parts or at the point of attachment of one part or an¬ other. Other studies also had shown that transverse cotyledon cracks re¬ duced germination, seedling growth and yield (Atkins, 1958 ; Waters and Atkins, 1959).

In an attempt to reduce seedling growth rate differences, we soaked seeds and removed seed coats after various stages of imbibition in shal¬ low water and observed further growth stages after planting in sand and peat mixtures. Transverse coty¬ ledon cracks were observed more fre¬ quently after soaking. When coty¬ ledons from swollen seeds were ex¬ amined before planting, it was ob¬ vious that the cracks had occurred during imbibition and not after

[43]

44

Transactions Illinois Academy of Science

planting. Individual cotyledons on wet filter paper (either with lower or upper epidermis in contact with the water) developed crescent cracks, beginning in the boundaries of the wet and dry tissue. Complete frac¬ tures often occurred.

We did find literature to suggest that seeds of beans, peas, and corn were damaged due to differences in seed coat permeability, rate of water imbibition, and uneven swelling (Mc¬ Collum, 1953; Shull and Shull, 1932). Resuhr (1941) described in¬ jury to soybean seeds due to 24 hours of soaking, the injuries being spot¬ ting and split cotyledons. The pur¬ pose of this paper is to present se¬ lected data from a number of similar studies designed to test the hypothe¬ sis that the transverse and crescent cracks (partial or complete frac¬ tures) occur in soybean seeds during imbibition.

Materials and Methods

In each of six replicates, 30 seeds of variety Wayne and Shelby were selected for uniform size and absence of visible damage. Each 30-seed sam¬ ple was divided into two groups of 15 seeds, one group being placed in a 9 cm Petri dish containing a 9 cm filter paper and 15 ml of water (room temperature) for 30 minutes of soaking. The seeds of both groups were planted at a depth of 5 cm in unsterilized sand and peat (equal volume, 6" diameter plastic pots) watered to saturation. Seedlings grew at room temperature for 14 days.

One four-replicate study of the ef¬ fect of temperature was conducted using seeds of Wayne. Sixty seeds, four Petri dishes with filter paper, and 100 ml of tap water were placed in incubators at 10, 20, 30, 40, 50, and 60° C. When the water reached

the desired temperature, 15 seeds and 10 ml of water were placed in each Petri dish and the swollen seed examined 12 hours later.

Results and Discussion

Typical results from one replicate are presented for the study of water soaking effects on planted seeds. Five days after planting, all of the 15 un¬ treated seeds of Shelby and 14 of Wayne had emerged. The last seed¬ ling of Wayne emerged on the sev¬ enth day. Of these, one cotyledon of Shelby and four of Wayne show¬ ed some small transverse cracks. On the eleventh day after planting, seed¬ lings were uniform in height in both varieties.

Of the seed soaked in water before planting, seven seeds of Shelby and seven of Wayne had emerged five days after planting. Five addition¬ al seedlings of Shelby and four of Wayne emerged on the seventh day. On the ninth day, another seedling of Wayne emerged. None of the other seeds germinated in either variety. Of the 12 seedlings of Wayne, 22 cotyledons were counted, 14 with one or more cracks. Two seedlings had one cotyledon as a re¬ sult of complete fracture near the point of attachment to the hypo- cotyl. Only three seedlings had both cotyledons without cracks. Of the 12 seedlings of Shelby, 24 cotyledons were counted, all with one or more cracks. On the eleventh day after planting, seedlings of both varieties were irregular in height. The non- emerged seeds of both varieties had severe cracks or complete fractures near the plumule.

Each experiment with our seeds of these varieties resulted in more coty¬ ledons of Shelbv with cracks than

*/

those of Wayne. We would prefer more trials with several sources of

Liu & Pappelis — Soybean Cotyledons

45

seed before suggesting that this is due to varietal differences to coty¬ ledon cracking. We conclude that uneven swelling caused a small per¬ centage of seedling damage in our previous experiments. Whether these cracks predispose the seedlings to pathogens or alters the cell death pat¬ tern in pith tissue remains to be studied.

The amount of damage to seeds of Wayne (average of four replicates) during 12 hours of soaking in water at various temperatures was as fol¬ lows : MFC, 100% of the seeds with one or both cotyledons cracked ; 20°C, 92%; 30°C, 93%; 40°C, 80%; 50°C, 55%; and 60°C, 19%. These observations are in agreement with those of McCollum (1953) for snap beans germinating in water or soil; as temperature increases (from 10 to 30°C), cracking decreases, es¬ pecially in soil conditions. We be¬ lieve, as did Shull and Shull (1932) and McCollum (1953) for their seeds, that cracks in soybeans occur simultaneously with water uptake as a result of uneven swelling produc¬ ing a tension crack in the dry inter¬ ior portion or along the boundry be¬ tween wet and dry tissue. It may be that the rate of penetration of water into the seed tissue at higher temperatures creates moisture gradi¬ ents less conducive to cracking. Or- plianos and Hey decker (1968) re¬ ported that oxygen deficiency in the interior of soaked snap beans re¬ sulted in injury and killing. Al¬ though we did not see ungerminated seeds without severe cracks or com¬ plete fractures in cotyledons, it may be that similar oxygen deficiencies exist in soybeans also under pro¬ longed soaking. Further study on this problem is required to discover

the extent of seed damage due to soaking in wet fields and the rela¬ tionship of this type of seed dam¬ age to vigor, disease resistance, and }deld.

Literature Cited

Atkins, J. S. 1958. Relative suscepti¬ bility of snap bean varieties to mechani¬ cal injury of seed. Proc. Amer. Soc. Hort. Sci. 72:370-373.

Bainer, R. and H. A. Borthwick. 1934. Thresher and other mechanical injury to seed beans of the lima type. Calif. Agr. Expt. Sta. Bull. 580. 30 p.

H umphrey, L. M. 1958. Soybean ger¬ mination. The effect of cleaning. Soy¬ bean Digest 18 (10): 22.

Liu, K. C. 1966. Death of cells in pith tissue of soybean seedlings. M. S. Thesis. Southern Illinois University. 32 p. McCollum, J. P. 1953. Factors affecting cotyledonal cracking during the germi¬ nation of beans ( Phaseolus vulgaris ). Plant Physio. 28:267-274.

Metzer, R. B. 1967. Natural and in¬ duced variations in soybean seed quality during maturation. Ph.D. Thesis. Iowa State University Library. 117 p.

Moore, R. P. 1957. Rough harvesting methods kill soybean seeds. Soybean Digest 17(4): 14-16.

Orphanos, P. I. and W. Heydecker. 1968. On the nature of the soaking in¬ jury of Phaseolus vulgaris seeds. Jour. Exp. Botany 19:770-784.

Resuhr, B. 1941. uber die bedeutung konstitutioneller miingel fur das auftrent- en von keimlingsschuden bei soja hispida moench. II. Zeitschr. Pflanzenkrankh. 51:161-192. (Biol. Abstr. 23:2850, No. 27499, 1949).

Shull, C. A. and S. P. Shull. 1932. Irregularities in the rate of absorption by dry plant tissue. Bot. Gaz. 93: 376- 399.

Tachibana, H., R. B. Metzer, and D. F. Grabe. 1968. Cotyledon necrosis in soybean. Plant Dis. Reptr. 52 : 459- 462.

Waters, E. C. and J. D. Atkins. 1959. Performance of snap bean ( Phaseolus vulgaris) seedlings having transversely broken cotyledons. Proc. Amer. Soc. of Hort. Sci. 74: 591-596.

Manuscript received November 13, 1970

A PROPOSED BIOCHEMICAL MECHANISM OF THE TOXIC ACTION OF DDT

ROBERT C. HILTIBRAN

Illinois Natural History Survey, Urbana, Illinois 61801

Abstract. — DDT, 5.9 x 10~4 grams per ml of reaction medium, inhibited oxy¬ gen uptake by bluegill liver mitochondria in the presence of succinic acid. DDT increased the hydrolysis of adenosinetri- phosphate in the presence of magnesium and manganese ions. A biochemical mech¬ anism of the toxic action of DDT is sug¬ gested.

DDT (1,1,1, -trichloro -2,2 bis (P- chlorophenyl) ethane) has been wide¬ ly used as an insecticide and appar¬ ently has spread throughout the world. DDT has been the subject of much research in efforts to explain its mode of action on insects, the pri¬ mary target organisms. More recent research has attempted to explain its effects on non-target organisms, such as fish and birds. Metcalf (1955) discussed the mode of action of DDT, and later O’Brien (1967) in¬ dicated that after 12 years of in-

*/

tensive research the mechanism of the insecticidal properties of DDT has not been elucidated. However, the general agreement was that the primary target of DDT appeared to be the nervous systems of both verte¬ brates and invertebrates.

This view is untenable, as it does not explain the diverse biological effects, such as the effects on fish and bird reproduction, which have been shown to be related to or caused by DDT. However, some of the ob¬ served results, such as increased ac¬ tivity, may be due to the effects of DDT on the nervous system, but

there is evidence that DDT affects fundamental biochemical processes in other tissues. The involvement of the production of cellular energy and the effects of DDT on these proces¬ ses was dismissed by Metcalf (1955) as of little importance in explain¬ ing the mode of the toxic action of DDT. The suggested effects of DDT on the nervous systems, extensively reviewed by O’Brien (1967) does not offer a suitable explanation of the effects of DDT on the reproductive processes in birds and fishes, while the effects of DDT on basic bio¬ chemical enzymatic processes may explain both phenomena.

We have been investigating the ef¬ fects of possible pollutants includ¬ ing pesticides on energy production by the mitochondria of the liver of the bluegill sunfisli, Lepomis macro- chirtiSf (Hiltibran, 1967b) and have found that compounds which were very toxic to fishes, such as rotenone, antimycin A, cyanide, isopropyl ester of 2,4-D altered the oxygen or phos¬ phate uptake by bluegill liver mito¬ chondria (Hiltibran, 1967b). The previous investigations of Sacktor (1950), Johnston (1951), and An¬ derson et al (1954) indicated that DDT altered the oxygen uptake by various tissues. The results of our investigation of the effects of DDT on the oxygen and phosphate metab¬ olism of bluegill liver mitochondria is reported.

[46]

Hiltibran — Biochemical Action of DDT

47

Methods

Native, wild bluegills were held in the laboratory aerated aquaria at 25° C and all enzymatic assays were conducted at that temperature. Pro¬ cedures for the preparation of the mitochondria, for estimating the oxy¬ gen and phosphate, for estimating the rate of release of inorganic phos¬ phate from adenosinetriphosphate, ATP, and for estimating the nitro¬ gen content of the mitochondrial preparations have been previously reported (Hiltibran and Johnson, 1965). The data are the average changes observed from three or more experiments and have been corrected for endogenous activity and the ef¬ fects of the solvent. Reference sam¬ ples of DDT were used, and redis¬ tilled ethyl alcohol or acetone were used as solvents. DDT concentra¬ tions are expressed as grams of DDT per ml of reaction medium.

Results

The effects of DDT on oxygen and phosphate uptake by bluegill liver mitochondria in the presence of suc¬ cinate and alpha-ketoglutarate as substrates are summarized in Table 1. In the presence of succinate, 5.9 x 10'4 g of DDT per ml of reaction medium completely inhibited the up¬ take of oxygen, and at a concentra¬ tion of 5.9 x 10"6 g oxygen uptake was inhibited approximately 40 per¬ cent. Lower levels of DDT were not as effective on oxygen uptake. Us¬ ually when there is severe inhibition of oxygen uptake there is an increase in the inorganic phosphate content of the reaction medium. However, in the presence of DDT there was not an increase in the inorganic phos¬ phate content of the reaction medi¬ um, when oxygen uptake is altered.

DDT inhibited oxygen uptake in the presence of alpha-ketoglutarate

Table 1. — Effects of DDT on Oxygen Uptake by Bluegill Liver Mitochondria.

g/ml of

Reaction Medium

Average

Change in n 102/hr/ mg N

Average

Change in

H moles POUhr/mg N

Percent Inhibition of O2 Uptake

Succinate

5.9 x 10~4 .

(-)

118

(±)

16

100

5.9 x 10-5 .

(-)

89

(±)

18/

80

5.9 x 10"6 .

(-)

39

(±)

9

40

5.9 x 10-8 .

(±)

30

(±)

4

Alph

a-Ketoglutarate

5.9 x 10 4 .

(-)

31

( + )

8

20

5 . 9 x 10~5 .

(-)

58

( + )

20

45

5.9 x 10 6 .

(-)

37

( + )

13

25

5.9 x 10 7 .

(+)

41

( + )

3

30

(increase)

48

Transactions Illinois Academy of Science

as substrate, but to a lesser extent. There was no effect of DDT on phos¬ phate uptake in the presence of al- pha-ketoglutarate. DDT did not ap¬ pear to uncouple the phosphate up¬ take from the oxidation of either substrate, therefore, the primary ef¬ fect of DDT appeared to be on the utilization of oxygen.

The effects of DDT on the hydroly¬ sis of ATP by the bluegill liver mito¬ chondria are summarized in Table 2. In the presence of cadmium, DDT inhibited the hydrolysis of ATP at all concentrations of DDT used, and in the presence of zinc, DDT inhib¬ ited the hydrolysis of ATP at the highest concentration of DDT. The hydrolysis of ATP was not greatly altered in the presence of either manganese or calcium, but in the presence of magnesium, DDT in¬ creased the hydrolysis of ATP from approximately 60 percent at a DDT concentration of 1.5 umoles of DDT per ml of reaction medium to 150 percent at a DDT concentration of 2.5 umoles.

The data suggests that DDT can alter the activities of various enzyme complexes from the bluegill liver mitochondria and that the observed

effects appear to be related to the concentration of DDT. The data also suggests a specific effect of the DDT molecule on each liver enzyme com¬ plex. When the DDT concentration was increased, complete inhibition of oxygen uptake occurred and this is consistent with the report that as the DDT poisoning of insects in¬ creased, the oxygen utilization de¬ creased, until the death of the or¬ ganisms occurred.

Discussion

Soon after the large scale use of DDT for the control of insect pests was begun, it became evident that bird populations (Robbins et al, 1951) declined in the DDT treated areas. Some of the observed changes appeared to be due to the effects of DDT on the viability and hatchabil- ity of eggs (Mitchell et al, 1951). More recent data have suggested that the wide-spread use of DDT and other organochloro insecticides may be responsible for the decline of pop¬ ulations of the golden eagle, Aquila chrysaetas (Lockie and Ratcliff e, 1964), Osprey, Pandion haliaetus (Ames, 1966), herring gull, Larus

Table 2. — Effects of DDT on Hydrolysis of ATP.

Metal

g of DDT per ml reaction medium

5.3 x 10"4

8.9 x 10-4

17.7 x 10-4

Average Change in n moles ATP/hr/mg N

Cadmium .

(-) 34

(±) 13 (+) 40

(+) 16 (±) 7

(-) 16 (±) 9

(+) 45 (+) 36

(±) 11

(-) 7

(—) 19

(±) 19 (+) 23

(±) 33

Zinc .

Manganese .

Magnesium .

Calcium .

Hiltibran — Biochemical Action of DDT

49

argentatus (Paynter, 1949), and the peregrine falcon, Falco peregrinus (Hickey and Anderson, 1968).

Lake trout ( Salvelinus namay- cush) sac fry, hatching from eggs of females from Lake George, New York, did not survive. However, when the sperm from males from Lake George were used to fertilize eggs of females from another water¬ shed, fry survival was normal. The watershed of Lake George had re¬ ceived applications of DDT for the control of gypsy moth. Burdick et al (1964) investigated this phenome¬ non, described the syndrome pro¬ duced in the lake trout sac fry, and pinpointed the time of its develop¬ ment as late in the yolk-sac utiliza¬ tion period and just before the fry began to feed. This time appeared to be correlated with the period of maximum utilization of the phospho¬ lipid content of the yolk-sac. Fur¬ ther, comparison of the affected fry with normal fry did not reveal any histological or pathological differ¬ ences. They noted that fry hatched from eggs which contained 2.95 ppm of DDT developed the syndrome, whereas fry from eggs with a DDT content of 2.67 ppm did not.

Allison et al (1963) reported that mortality among sac fry appeared to be highest in those cutthroat trout ( Salmo clarki lewisi) which received high concentrations of DDT. Maeck (1968) reported similar observa¬ tions with brook trout ( Salvelinus fontinalis) which had received re¬ peated sub-lethal concentrations of DDT.

The recent work reported by Ames (1966) on the Osprey in Connecti¬ cut and the work of Paynter (1949) on the herring gull in Lake Michi¬ gan indicated that DDT is the causa¬ tive agent in the decrease in popu¬ lations of these two species and that the effect was on the development

of the embryo. It has been suggested that in the eagle (Lockie and Rat¬ cliff e, 1964) and peregrine falcon (Hickey and Anderson, 1968) DDT caused a reduction in egg shell thick¬ ness. However, the effects of DDT on the reproduction of the quail (Dewitt, 1956) and the pheasant ( Phasianus colchicus ) (Genelly and Rudd, 1956) do not fit this pattern, for their chicks die several weeks after hatching.

DDT can cause mortality among fishes and birds. However, repeated sub-lethal doses of DDT did not cause mortality. Further, sub-lethal doses of DDT to fishes and birds did not cause any reduction in the number of viable eggs produced. In fishes, normal sac-fry were produced from females from the Lake George water¬ shed which apparently were not sus¬ ceptible to the effects of DDT until late in the sac-fry stage. The fish af¬ fected during sac-fry stages did not show any histological or pathological damage. Bird embryos, however, ap¬ peared to die in embryonic life. Thus, it would appear that DDT must be altering some fundamental biochemical or physiological process¬ es common to all the organisms and to the various situations discussed. Energy production would be such a common denominator.

The increased oxygen consumption and the associated random, awkward body movements observed in insects treated with DDT suggest a neuro¬ muscular involvement. It is gener¬ ally agreed that the increased oxygen consumption is the result of increased activity of the organisms (Metcalf, 1955, O’Brien, 1967). However, Riker (1946), Jandorf et al (1946), and Laug and Fitzhugh (1946) found that the oxygen consumption of iiver slices of animals in sub-acute and chronic DDT poisoning had an increased oxygen consumption, and

50

Transactions Illmois Academy of Science

that the oxygen consumption from animals in advance stages of DDT poisoning was decreased. The changes in body coordination could be considered as a partial impair¬ ment of functions due to the de¬ creased ability of the nervous system to function normally, a possible re¬ sult of reduced energy output.

Part of the reaction of the or¬ ganisms would appear to be a re¬ sponse to stress. It was observed that as the poison symptoms became more severe, oxygen uptake declined (O’Brien, 1967), which is consistent with the observed effects of DDT on oxygen uptake as described above. The first system to be involved might be the nervous system because of its high fat content. This point will be discussed later.

Rotenone, which has been used as an insecticide but more recently has been widely used as a fish toxicant, has been shown to decrease oxygen uptake by insects (Tishler, 1935) and was thought to cause damage to the tissues (Danneel 1933). Fu- kami and Tomizawa (1956) reported that rotenone inhibited the oxidation of glutamate, and later it was shown that this compound did not cause tissue damage (Oberg, 1955) but that its primary effect was the in¬ hibition of the transfer of electrons from the substrate to the cytochrome chain (Lindahl and Oberg, 1961). This has been the only biochemical effect shown for rotenone. It should be pointed out that in rotenone poi¬ soning, excessive, random movements of fishes are observed, but rotenone has not been considered a nerve poi¬ son (O’Brien, 1967). The effects of rotenone on the oxygen uptake by bluegill liver mitochondria have been reported (Hiltibran and Johnson, 1965).

Cyanide and antimycin A have been used as fish toxicants (Bennett,

1962), (Walker et al, 1964) and ap¬ pear to block the uptake of oxygen by bluegill liver mitochondrial sys¬ tems (Hiltibran, 1965, 1967a). They interrupt the electron flow from the substrate to the electron acceptor, oxygen (Chance, 1956). The data strongly suggest that when the oxi¬ dative pathways are blocked, energy cannot be produced, various cells cannot function, and if some regula¬ tory cells or organ would be affected, for example the respiratory center, the organism cannot maintain its integrity and dies. This has been suggested previously (Hiltibran, 1971), (Skidmore, 1965).

It is well known that some metals are extremely toxic to bluegills (Mc¬ Kee and Wolf, 1963). Recently we observed that cadmium and zinc at relatively low levels (Hiltibran, 1965; 1967a) severely inhibited the uptake of oxygen by bluegill liver mitochondria. We found also that calcium and manganese can inter¬ rupt energy production, but the ef¬ fect was primarily on phosphate metabolism, whereas the primary ef¬ fect of cadmium and zinc was on the uptake of oxygen (Hiltibran, 1971).

Previously we had been intrigued by the fact that certain derivatives of 2,4-D were more toxic to small bluegills than were others (Hilti¬ bran, 1967c). Therefore, we investi¬ gated the effects of nine derivatives of 2,4-D and found that the most toxic 2,4-D derivatives used in the study, the butyl and isopropyl es¬ ters, altered the uptake of oxygen and phosphate by the bluegill liver mitochondria. Thus it would appear that the primary effect of these de¬ rivatives was on oxygen uptake and that their toxic action could be pro¬ duced via their effects on energy production (Hiltibran, 1969a; 1969b). It appears, then, that the

Hiltibran — Biochemical Action of DDT

51

effects of DDT on the oxygen uptake cited above would be of paramount importance in explaining the mode of action of DDT, since we have shown the similarity of the effects of DDT, rotenone, cadmium, zinc, the butyl and isopropyl esters of 2,4-D, and other electron flow inhibi¬ tors and oxidative phosphorylation uncoupling agents.

Thus I believe that the interrup¬ tion of the production of energy is of primary importance in explaining the toxic action of DDT. I suspect that similar cases can be developed for some of the other organochloro insecticides on the basis of available data ( Hiltibran unpublished data ; Colvin and Phillips, 1968).

DDT is very soluable in or has a high affinity for the tissue fat. Fur¬ ther, it is assumed that the incor¬ poration of DDT into the tissue fat is a passive process and is not an active “detoxification” process. It does not appear that definitive data are available to determine whether this incorporation is a biological ac¬ tive or a biological passive process.

Exposure of target or non-tar- get organisms to lethal levels of DDT would result in a DDT up¬ take which would exceed the rate at which DDT could be incorpor¬ ated into the tissue fat and/or ex¬ ceed the DDT storage capacity of the total body fat content. This amount of DDT would be in addi¬ tion to that which would be metab¬ olized. The remaining “unbound DDT” or “circulating DDT” would be able to exert its toxic action by blocking the oxygen uptake in the various tissues. Repeated light doses of DDT to fishes and birds would give time for the storage of the DDT in the tissue fat, and this could account for the large buildup of DDT without apparent damage.

Apparently the DDT is transfer¬

red to the eggs of fishes or birds which would account for the high levels of DDT in these eggs. In fishes, most of the DDT would re¬ main bound throughout the devel¬ opment of the fish embryo, and DDT conld not exert its toxic action un¬ til the late stage of the development when the yolk fat was mobilized, and the DDT again became “cir¬ culating” or “unbound”. If only small amounts of DDT were in¬ volved, the effects on fry would not be lethal. This is consistent with the data in Table 1 and with the ob¬ servations of Burdick et al (1954). The DDT content of bird eggs which did not develop normal embryos has not been as well documented as has the DDT content of fish eggs ; the effects of DDT on bird reproduction is not as clearly defined as with the fishes. However, the developing bird embryos apparently were severely af¬ fected by high levels of “circulat¬ ing DDT”.

There are intraspecies differences in the quail and pheasant that com¬ plicate the picture, and these will remain a mystery until the com¬ parative biochemistry of the organ¬ isms involved is known. These dif¬ ferences might help to explain why quail and pheasant chicks are the susceptible unit to the lethal effects of DDT. It is suspected that the “circulating DDT” is the causative agent, but apparently the stress de¬ velops later which may coincide with the production of feathers or using the remaining portion of the yolk, which may contain large quantities of DDT.

Data to support this hypothesis comes from the work of Johnston (1951), Anderson (1954), Riker (1946), Jandorf et al (1946), and Laug and Fitzhugh (1946). Judah ( 1949) could not demonstrate any ef¬ fect of DDT on the succinic oxidase,

52

Transactions Illinois Academy of Science

but Johnston conlcl. Judah had ad¬ ministered the DDT in his experi¬ ments in an oil emulsion, whereas Johnston had administered the DDT in acetone. Further, when Johnston administered DDT in an oil emulsion, he could not demonstrate any effect of DDT on the succinic oxidase. This demonstrates the protective effect of the oil or fat on the DDT. It is not surprising, therefore, that with the high fat and phospholipid content of the nervous tissue that nervous systems might be one of the first tis¬ sues affected.

It has been suggested that DDT altered the production of the egg shell by the peregrine falcon (Hic¬ key and Anderson, 1968) and the golden eagle (Lockie and Ratcliff, 1964), and this effect of DDT was confirmed in experiments with the American sparrow hawk, Falco spar- verius (Porter and Wiemeyer, 1969). The synthetic formation of the egg shell, primarily calcium carbonate, would suggest that its synthesis is an energy-requiring process for the mobilization of the required calcium and for the production of the car¬ bonate. Torda and Wolff (1959) demonstrated that DDT inhibited the carbonic anhydrase enzyme com¬ plex, which has been suggested as one enzyme complex involved in the formation of the egg shell (Sturkie, 1954). Further, the effects of organ- ochloro pesticides on the metabolism of endogenous steroids is not known (Kupfer, 1967), but recently Welch et al (1969) reported that a DDT isomer was converted to an estro- gentic hormone metabolite which would indicate that organochoro in¬ secticides, particularly DDT, may have some effect on the steroid hor¬ mones involved in calcium metab¬ olism.

Johnston (1951) found that DDT did not alter the succinic dehy¬

drogenase but that the succinic oxi¬ dase complex was affected. The data in Table 2 indicate that DDT did not appreciably alter the oxidation of alpha-ketoglutarate. These ob¬ servations indicate that DDT in¬ hibited the flow of electrons at the flavoprotein transferring site be¬ tween the succinic dehydrogenase and the cytochrome chain. Matsu- mura and O’Brien (1966) have iso¬ lated complexes of DDT and tissue components and suggested that the complexes involved were of the charge-transfer type. These data would further support the biochem¬ ical hypothesis suggested, since DDT could form a complex with the flavo¬ protein of the electron transferring site between succinic acid and the cytochrome chain. Such action could block the flow of electrons, which would block the production of en¬ ergy via the oxidative pathways. DDT has been found to have an ef¬ fect similar to rotenone, cyanide, and antimycin A, and it is suggest¬ ed that this is the primary effect of DDT. DDT also has been shown to have an effect on phosphate metab¬ olism, as indicated by its effect on the hydrolysis of ATP, which ro¬ tenone, cyanide, and antimycin A did not alter. The data suggest that the primary effect is the inhibition of electron flow from succinic acid to the cytochrome chain.

Acknowledgments

The author acknowledges the technical assistance of Messrs. Michael Johnson and Richard Schumacher and Mrs. Charlene Mathis during this investigation.

Literature Cited

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Repeated Exposure to DDT. Science 142: 958-961.

Hiltibran — Biochemical Action of DDT

53

Anderson, A., R. March, and R. L. Met¬ calf. 1954. Inhibition of the succin- oxidase system of susceptible and resistant house flies by DDT and related com¬ pounds. Ann. Entomol. Soc. 47 : 595- 601.

Ames, P. S. 1966. DDT Residues in the eggs of the Osprey in the North-Eastern United States and their Relation to Nesting Success. J. Applied Ecol. Sup-

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Bennett, G. W. 1962. Management of Artificial Lakes and Ponds, Reinhold Publishing Co. N.Y. p. 142-149.

Burdick, G. E., E. J. Harris, H. J. Dean, T. M. Walker, J. Shea, and D. Colby. 1964. The Accumulation of DDT in Lake Trout and the Effect on Reproduc¬ tion. Trans. Amer. Fish. Soc. 93: 127- 136.

Chance, B. 1956. “Enzymes: Units of Biological Structure and Function.” Academic Press, Inc. New York. p. 450.

Colvin, H. J., and A. T. Phillips. 1968. Inhibition of Electron Transport En¬ zymes and Cholinesterases by Endrin Bull. Environmental Contamination and Toxicology 3 : 106-115.

Danneel, R. 1933. Die Giftwirkung des rotenons and seinen Derivativ anf fische. Ill Der Angriffspunkt der Gifte. Zeit. vergl. Physiol. 18: 524-535.

Dewitt, J. B. 1956. Chronic toxicity to Quail and Pheasants of some Chlorinated Insecticides. J. Agr. and Food Chem. 10: 863-866.

Fukami, J., and C. Tomizawa. 1956. Ef¬ fects of rotenone on the 1-glutamic oxi¬ dase systems in insects. Japanese Contri¬ butions to the Study of the Insectcide- Resistance Problem, p. 212-216. Orig¬ inally published by Boytu Kagaku 2 1 : 129-133.

Genelly, R. E., and R. L. Rudd. 1956. Effects of DDT, Toxaphene and Diel- drin on Pheasant Reproduction. The Auk 73: 529-539.

Hickey, J. J., and D. W. Anderson. 1968. Chlorinated Hydrocarbons and eggshell changes in raptorial and fish-eating birds. Science 162: 271-273.

Hiltibran, R. C. 1965. Oxidation of

succinate by bluegill liver mitochondria. Trans. Ill. Acad. Sci. 58: 176-182.

Hiltibran, R. C. 1966. Hydrolysis of

adenosinetriphosphate by bluegill liver mitochondria. Trans. Ill. Acad. Sci. 59: 249-253.

Hiltibran, R. C. 1967a. Oxidation of

alpha-ketoglutarate by the bluegill liver mitochondria. Trans. Ill. Acad. Sci. 60: 244-249.

Hiltibran, R. C. 1967b. Abstracts. 7th International Congress of Biochemistry. Tokyo, Japan, p. 899.

Hiltibran, R. C. 1967c. Effects of some herbicides on fertilized eggs and fry. Trans. Am. Fish. Soc. 96: 414-416.

Hiltibran, R. C. 1969a. The hydrolysis of adenosinetriphosphate by bluegill liver mitochondria in the presence of 2,4- dichlorophenoxyacetic acid derivatives. Trans. Ill. Acad. Sci. 62: 36-43.

Hiltibran, R. C. 1969b. Oxygen and phosphate metabolism of bluegill liver mitochondria in the presence of 2,4-di- chlorophenoxyacetic acid derivatives. Trans. Ill. Acad. Sci. 62: 175-180.

Hiltibran, R. C. 1971. Effects of cad¬ mium, zinc, manganese and calcium on the oxygen and phosphate metabolism of bluegill liver mitochondria. J. Water Poll. Contr. Fed. In press.

Hiltibran, R. C., and M. G. Johnson. 1965. The effect of rotenone on oxygen uptake by liver mitochondria of the bluegill, Lepomis macho chirus. Trans. Ill. Acad. Sci. 58: 140-143.

Jandorf, B. J., H. B. Sarett, and O. Bo- dansky. 1946. Effect of oral adminis¬ tration of DDT on the metabolism of glucose and pyruvic acid in rat tissues. J. Pharmacol, and Exp. Therap. 88: 333- 337.

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Macek, K. J. 1968. Reproduction in Brook Trout ( Salvelinus pontinalis) fed sublethal concentrations of DDT. J. Fish. Res. Bd. of Canada. 25: 1787-

1796.

Matsumura, F., and R. C. O’Brien. 1966.

54

Transactions Illinois Academy of Science

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166.

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Manuscript received May 4, 1970

SURFACE TENSIONS OF BINARY SOLUTIONS OF NITROPARAFFINS IN CARBON TETRACHLORIDE

CLAUDE R. GUNTER, RICHARD D. MADDING, JR.

THOMAS E. HANSON, AND BORIS MUSULIN Department of Chemistry, Southern Illinois University, Carhondale, Illinois 62901

Abstract. — The surface tensions of binary solutions of nitromethane in carbon tetrachloride and nitroethane in carbon tet¬ rachloride were determined by the ring me¬ thod at 30°, 35°, and 45 °C. Assuming perfect solutions, the surface excesses and the surface entropies and enthalpies were calculated. The surface entropies indicat¬ ed increasing ordering in the solutions. Surface entropies and enthalpies of mixing were also calculated. These calculations indicated that nitroethane formed a more ideal solution than nitromethane. Nitro¬ methane showed evidence of association as well as disassociation. The deviations for these solutions are within the limits of an empirical mixture law.

This investigation is a continua¬ tion of the series of simple physi¬ cal measurements upon binary solu¬ tions of nitroparaffins in carbon tetrachloride. The immediate pur¬ pose of such measurements is to place into the literature accurate solution data concerning commercial chemicals that have important ap¬ plications in automotive fuels. Al¬ though data involving pure nitro¬ paraffins, particularly nitromethane is plentiful, solution data is sparse (cf. Timmermans (1959)). The cur¬ rent series of papers indicates the degree to which solution values, for engineering purposes, can be obtain¬ ed by interpolation and/or extrapo¬ lation of existing data and by pre¬ diction from empirical formulae.

A second major objective of the current investigations is to obtain

inferential information concerning the structure of these binary solu¬ tions. First, are the solutions ideal, regular, or irregular? Second, if the solutions are irregular, what is the nature of the irregularity? Gunter et al. (1967) by means of density measurements have shown that these binary solutions are irregular with respect to volume effects. Wettaw et al. (1969) by means of viscosity measurements have shown that these binary solutions are irregular with respect to entropy effects. The pres¬ ent paper shows the nature of ir¬ regularities determined by surface tension measurements. Since infer¬ ences concerning the bulk solution may differ from inferences concern¬ ing the surface layer, this work has the advantage of providing two sets of conclusions with one set of meas¬ urements. Third, how do solutions of homologous members of the nitro- paraffin series differ? The previous sets of measurements indicated that the first member of the series is quite different from the second member. Whether or not the same conclusion is true for each different physical property is determined only by mak¬ ing the measurement.

This paper discusses the surface tension, the surface excess, the sur¬ face entropy and enthalpy, and the excess surface entropy and enthalpy

[55]

56

Transactions Illinois Academy of Science

of mixing as functions of concen¬ trations, and their relation to the molecular structure of the pure nitroparaffins. In order to acquire a proper perspective between older and more modern investigations of regularity, one empirical relation¬ ship, the parachor, is examined.

Experimental

Fisher certified and spectro grades of carbon tetrachloride, Fisher cer¬ tified grade nitromethane, high pur¬ ity research samples of nitromethane and nitroethane (Commercial Sol¬ vents), and highest purity nitro¬ ethane (Brothers Chemical Co.) were used without further purifica¬ tion. The solutions were prepared by mixing volumes of pure com¬ ponents. The estimated cumulative transfer error was ±0.005 mole frac¬ tion.

The solutions, in closed, ground glass containers, were brought to equilibrium in a Precision Scien¬ tific Co. bath (No. 66580) and regu¬ lated (±0.02cC.) with a Merc to Merc Modi1! PS-62510-D1 thermoreg¬ ulator. Temperature readings, pre¬ cise to ±0.01 C., were obtained with a thermometer calibrated with a Na¬ tional Bureau of Standards ther¬ mometer. The precision in the prep¬ aration of solutions dictated round¬ ing of the temperature readings by 0.1 °C. or less to integral values.

The apparent surface tensions were obtained, after rapid transfer of the solutions from the bath, bv the ring method using a Fisher Sur¬ face Tensiometer, Model 20. True surface tensions were calculated with the formula for the correction fac¬ tor.

where DAv is the average dial read¬ ing; y is the true surface tension; R wire and R,.infr are the radii of the suspending wire and the platinum ring, respectively ; C is the circum¬ ference of the ring ; and DHq and Dvap are the densities of the solu¬ tion and the vapor in equilibrium with the solution, respectively. Va¬ por densities were estimated, by linear interpolation, for use in the correction calculation ; liquid densi¬ ties were taken directly from or es¬ timated from Gunter et al. (1967).

Results

The number of readings and the average deviation of each set of read¬ ings are given in Table 1. These averages sometimes represent read¬ ings taken on different davs ; some- times they represent readings taken on different sample grades ; and sometimes they represent readings on repetitive trials taken to ascer¬ tain temperature loss in the trans¬ fer process. There is no explanation for the readings with the greatest deviation, i.e. 0.8 and 0.9 CH3 N02, except lack of experimental tech¬ nique. Nevertheless, even these read¬ ings are acceptable for an instru¬ ment whose scale gradations are giv¬ en in 0.1 units. Each set of readings was processed by a computer pro¬ gram which calculated the average deviation and the probable error of an individual reading, tested each individual reading for discard as being beyond normal experimental error, and, if necessary, recalculated the averages and the deviations. The discard judgment was based on Chauvenet’s criterion (Worthing and Geffner [1943]). Insomuch as

.04534 - 1.679

,]

.01452 DAv

—

>1

( IW)_

+

_C-* (!>,,

- o . ) _

Table 1. — Surface Tensions of Carbon Tetrachloride-Nitroparaffin Solutions.

Gunter et al — Nitroparaffin Surface Tensions

57

Av.

Dev.

Dial

Read.

o

o

No.

Read.

i~n

Prob¬

able

Error

(dyne/

cm)

y

(dyne/

cm)

Av.

Dev.

Dial

Read.

u

o

No.

Read.

LO

CO

Prob¬

able

Error

(dyne/

cm)

y

(dyne/

cm)

Av.

Dev.

Dial

Read.

u

o

No.

Read.

O

co

Prob¬

able

Error

(dyne/

cm)

y

(dyne/

cm)

Mole Fraction

(RN02)

ID

C

rt

■M

V

O — ' O O — 'OOOOOO

VOON^^LOM(NNMNI?\

rH i—H t— H r- H r-H i-H r-H i-H i-H

rH r-H r-H i— H r-H r-H r-H r-H i— I CvJ

ooooooooooo

Lo tM in c^^Ot— 'ONChfNcnoOrHNONOON

■^•^^unLnm^O'ONCiN

CN(N<N(N(N(N(N(NCNNtn

TfiO^^OOO^N^HtNN

OOOOOi— 'OOomcmo

'v0O'-olol^>O1j^,-oOOcn

CM »—i »-h »— h CM r— H r— i — CM i-h

— 1 O CM r— ' i— IClTfr- <

OOOOOOOOO

NOrHO\(NOOOOST)lVOQ

NC\0(NiJiinONMTjiO

min'O'O'O'ONNONi-'-^

Cl M (N (N (N <N C l (N C) rn CM

OOOON

o o o

0\ c*-) r— i oo oc On c<c

o o o o o o o

OOOloOOOOOlo^o

TjirtrH^HtnciciNcirt' — i

CldrHrH

o o o o

r— I CM r-H I

o o o o

OoooOcunHdNOiO'^

NooOi-icninNcnNN^

'O'CNNNNNOOON'hio

CICICICICICICICINCjO

O’-idcn^m'ONOOO'O

dboobbdbdow

aj

c

ctS

_G

■(->

4J

o

■l-l

£

OMO'O't* OO N On VO Tt< 1^1 OO OOOOOOOOOOO

vOOnOnOnOnOnOnCnC\CnC\

CM

■CrHClr-lCMCICICIrHr-lCI

ooooooooooo

inONCM^OcnCNClr-iCNrH

\Ot^OO-— ilooOi— it^^CMr- 1

^^T*unmm'OiaNOOC\ Cl CM CM CM CM CM Cl CM CM CM CM

TfOMOQiNiCTticoO^i*

OOOOOOOOOOO

lO'ninimninininO'ci lo CM CM t— i CM i— ii— i I

OOOOOO

CM o I— I CM

o o o o

NNOO

NOOO

i >0 Tf CM i-H cm On MD I LO OO Tl"1 O MD CM 1— I

inin\0'O'O'ON00000\O

(Ncicicicicicicicicicn

ooincicncncN

OOOOOO

dCN'Om

O O O O

0L'">00000,-ciLn'-c~>0 -f CM >— i i— i i— l i— i CM

' o o ■

o o o o

O cm c<c N\ON

cn m \0 Oi t|i 'O \0 cn in Oi m cm oo \0

'O'OVONNNNOOONOnO

CM CM CM CM CM CM Cl Cl CM CM CO

Oi-lCMdV^imONOOONO

ddddddddbd— I

58

Transactions Illinois Academy of Science

this particular criterion was used for decision making, the standard deviation of each set of readings was not recovered in the computer out¬ put. The standard deviation, ascer¬ tained by trial calculations, is ap¬ proximately 1.1 to 1.2 the average deviations tabulated in Table 1.

The probable error of the true sur¬ face tension calculated with Equa¬ tion (1) was calculated in the usual formulas for propagation of errors (cf. Daniels ct al. (1962)) using the errors given by Gunter et al. (1967) for Dliq , the average deviations of DAv , and zero error in Rwire , Rring , C, and Dvap . The probable errors are given in Table 1 along with the true surface tensions calculated from Equation (1). The Dliq determina¬ tion involved the compounding of errors from several sources which makes it, relatively, less determined than DAv which involved a single, accurate measurement. The probable errors shown in Table 1 reflect the weighted importance of the errors in

■^liq

Individual measurements at 30° C. showed no significant variation for the different grades of carbon tetra¬ chloride and nitromethane. The re¬ sults are in good agreement with the nitroparaffin values interpolated from the ring method data of Boyd and Copeland (1942). The results are lower than the data measured by and/or the data calculated from the non-ring methods used by Snead and Clever (1962) and Thompson, Coleman, and Helm (1954). The greatest 30° deviation from the lit¬ erature involves CC14 and is prob¬ ably due to slight evaporation loss during transfer. On the other hand, the deviation found in C2H5N02 is undoubtedly due to the impurities inherent in the grade of chemical used. Although of superior quality, the C2H5N02 is not of the great

purity as the CH3N02 and CC14. No literature values were available at 35°C or 45°C. One may surmise that the present data are of quality comparable to that of the 30° C data with somewhat greater experimental error due to evaporation at the high¬ er temperature. A comparison of the rapid ring method with a more accurate technique, e.g. the maxi¬ mum bubble pressure method, is ob¬ tained by comparing the values from the literature found in Table 6. No literature data for the solutions are available but the present data can be construed to be of the same ex¬ cellent quality as the data for our pure compounds.

The relative surface adsorption (surface excess) of the nitroparaffin (component 2) with respect to the carbon tetrachloride (component 1),

r2>1 was calculated from the equa¬ tion for perfect solutions (Defay

et al. (1966) ),

r2,i = / 9 7 \ (2)

RT \92 DT-P

where x2 is the mole fraction of the nitroparaffin, R is the ideal gas con¬ stant, T is the absolute temperature, and P is the ambient pressure. The derivative in Equation (2) was ob¬ tained analytically from a least squares fit of the surface tension data to a quadratic function of the mole fraction. The choice of a quad¬ ratic function was arbitrary but lim¬ ited by the criteria of a good repre¬ sentation of the input data and statistical reliability for the number of degrees of freedom. The zero de¬ gree and first degree polynomials are eliminated by the first criterion while any polynomial of quintic de¬ gree or higher is eliminated by the second criterion. Consideration of the errors inherent in determining a derivative through the use of data fit plus the errors in assuming the

Gunter et al — Nitroparaffin Surface Tensions

59

validity of Equation (2) suggested that cubic degree or higher poly¬ nomials were an over determination. The absolute surface adsorption of the nitroparaffln, r2, was calculated assuming a dividing surface under an inhomogeneous monolayer and a mixture surface tension which is a linear function of the surface ad¬ sorption mole fraction, r2/2 + Ti), (van Rysselberghe (1938)). The re¬ sults are given in Table 2. In gen¬ eral, the results illustrate the prin¬

ciple that the substance of lower sur¬ face tension is concentrated at the surface. For example, the negative values of r2>1 reflect the increasing deficiency of RN02 with respect to CC14 (the substance of lower sur¬ face tension). The values of r2 are consistent with the size of the nitro¬ paraffin molecule. The amount of excess is proportional to the differ¬ ence in surface tensions of the com¬ ponents of the solution. The more negative values of r2<1 for CH3N02

Table 2. — Surface Adsorptions of Carbon Tetrachloride-Nitroparaffin Solutions.

Stated in Units of Q cm 2

o

o

r— H

r2,l

r2

Mole Fraction

(RN02)

30°C

35°C

45 °C

30°C

35°C

45 °C

Nitromethane

0.0 .

0.0

0.0

0.0

0.0

0.0

0.0

0.1 .

0.190

0.143

0.107

—0.0462

—0.0241

—0.0154

0.2 .

0.152

0.0776

0.334

—0.0340

—0.0107

—0.00601

0.3 .

—0.115

—0.196

—0.220

0.0159

0.0369

0.0470

0.4 .

—0.610

—0.678

—0.653

0.0836

0.0997

0.110

0.5 .

—1.33

—1.37

—1.27

0.152

0.149

0.254

0.6 .

—2.29

—2.27

—2.06

0.221

0.327

0.368

0.7 .

—3.47

—3.37

—3.03

0.392

0.545

0.537

0.8 .

—4.88

—4.69

—4.18

0.735

1.11

0.872

0.9 .

—6.51

—6.21

—5.51

1.18

2.06

1.37

1 0

—8 38

—7 94

—7.03

Nitroethane

0.0 .

0.0

0.0

0.0

0.0

0.0

0.0

0.1 .

—0.0219

—0.0416

—0.0313

—0.00316

0.0113

0.0121

0.2 .

—0.112

—0.147

—0.128

0.00451

0.0642

0.0379

0.3 .

—0.271

-0.317

—0.291

0.00989

0.207

0.129

0.4 .

—0.498

—0.550

—0.519

0.150

0.271

0.283

0.5 .

—0.794

—0.847

—0.814

0.301

0.405

0.455

0.6 .

—1.16

—1.21

—1.17

0.521

0.807

0.634

0.7 .

—1.59

—1.63

—1.60

1.02

1.32

0.947

0.8 .

—2.09

—2.12

—2.09

1.68

1.82

1.43

0.9 .

—2.66

—2.67

—2.65

2.08

2.98

2.15

1 0

—3 30

—3 29

—3.27

60

Transactions Illinois Academy of Science

mirror the greater differential in the y-values of CH3N02 and CC14 as compared to C2H5N02. Although the specific values of r2 depend upon the mode of calculation (the choice of surface), these qualitative con¬ clusions remain invariant (Guggen¬ heim and Adam (1933)).

The surface entropy per unit area, So-, was obtained from a least squares fit of surface tension data to a linear function of temperature. The linear function was chosen because of the well-known behavior of surface ten¬ sion as a function of temperature (Partington, 1951). This fortuitous functional behavior results in a par¬ ticularly simple calculation whereby the slope of the least squares fit is simply (dy/dT)PA (A being the sur¬ face area). The identification of this slope as sa results from the alter¬ nate view of surface tension as sur¬ face free energy and the application of the usual thermodynamic equa¬ tions to the surface layer. The heat of extension of the surface per unit area (latent heat per unit area), ir , was calculated from siCT and, then, the enthalpy of extension of the surface per unit area, h^ , was calculated from la and y. That is,

the latent heat is obtained bv a tern-

%/

perature multiplication of the slope of the least squares linear plot. The enthalpy then follows immediately from the first law of thermodynam¬ ics, that is, from the conservation of energy. The values of sCT and h^ are given in Table 3.

The surface entropy per mole, sm, was calculated from the slope of a Ramsay-Shields (1893) plot deter¬ mined by the method of least squares. This calculation proceeds exactly as that of the preceding paragraph but the independent variable is not y but y (MV) 2/3, where M is the molecular mass of the compound and V is the

molar volume of the compound. For solution data, M was obtained by the assumption that mass of the solution is additive with respect to the mole fraction of the components,

M = x1M1 + x2M2 (3)

where x4 and x2 are the mole frac¬ tions of CC14 and nitroparaffin, re¬ spectively ; and M2 are the mo¬ lecular masses of CC14 and nitro¬ paraffin, respectively ; and M is the molecular mass of the solution. The molar volumes were taken from Gun¬ ter et al. (1967). Whereas y is the surface free energy, the quantity y(MV)2/3 is the free molecular sur¬ face-energy. As is true for

y,y(MV)V»

fits well a linear function of tem¬ perature, if the temperature is given in absolute terms. In a manner anal¬ ogous to the calculation of la and h^ , the heat of extension of the sur¬ face per mole (latent heat per mole), lm, and the enthalpy of extension of the surface per mole, hm, were cal¬ culated from sm. The entropliies and enthalpies are tabulated in Ta¬ ble 3.

The values of sCT and h^ for pure nitroparaffins are m good agreement with those given by Snead and Clever (1962) while the pure CC14 values, like those of Vogel (1948) are high¬ er than most values which can be obtained from literature data. The fact, that the values for CH3N02 are slightly higher than the best litera¬ ture values while the C2H5N02 val¬ ues are equivalent to the best litera¬ ture values implies that the slope of the CH3NO._, y-t function is chang¬ ing more rapidly than usually ac¬ cepted ; the conclusion is not incon¬ sistent with the possibility of slight¬ ly more evaporative loss at higher temperatures for the lower boiling CH3N02. A greater variation is

Gunter et al — Nitroparaffin Surface Tensions

Table 3. — Surface Thermodynamic Quantities.

61

Enthalpy

Entropy

h^

(ergs/ cm

2)

hra x 10 10 (ergs/mole)

Mole

Fraction

(RN02)

Sc r

(ergs/cm2-deg)

sm x 10 7 (ergs/ mole-deg)

30 °C

35 °C

45 °C

30°C

35°C

45 °C

Nitromethane

0.0

0.133

20.0

66.9

66.7

66.9

10.8

10.8

10.8

0.1

0.137

20.3

68.4

68.2

68.4

10.8

10.8

10.8

0.2

0.137

19.6

68.6

68.2

68.5

10.5

10.4

10.5

0.3

0.124

16.6

64.8

64.6

64.8

9.46

9.44

9.46

0.4

0.128

16.5

66.3

66.0

66.2

9.33

9.28

9.32

0.5

0.107

11.8

59.8

59.4

59.7

7.76

7.68

7.75

0.6

0.103

10.4

58.9

58.8

58.9

7.19

7.17

7.19

0.7

0.107

10.1

60.8

60.8

60:8

7.05

7.03

7.04

0.8

0.120

11.1

66.0

66.2

66.0

7.34

7.36

7.34

0.9

0.133

12.5

72.0

72.4

72.1

7.85

7.89

7.87

1.0

0.172

17.8

87.7

87.1

87.6

9.71

9.64

9.70

Nitroethane

0.0

0.133

20.0

66.9

66.7

66.9

10.8

10.8

10.8

0.1

0.120

17.3

63.1

62.9

63.0

9.94

9.99

9.93

0.2

0.122

17.4

63.7

63.7

63.7

9.89

9.89

9.89

0.3

0.128

18.2

65.9

65.8

65.9

10.1

10.1

10.1

0.4

0.114

15.2

61.7

61.6

61.7

9.15

9.10

9.14

0.5

0.113

14.7

61.8

61.6

61.7

8.95

8.91

8.94

0.6

0.119

15.6

63.9

64.0

63.9

9.21

9.22

9.21

0.7

0.125

16.5

66.5

66.6

66.5

9.47

9.48

9.48

0.8

0.122

15.5

66.1

66.1

66.1

9.20

9.20

9.20

0.9

0.111

13.5

63.4

63.4

63.4

8.59

8.59

8.59

1.0

0.104

12.0

62.2

62.2

62.2

8.13

8.14

8.13

found for the molar thermodynamic quantities because of the variations in the density data. Both s^ and sm for nitromethane exhibit a mini¬ mum and, thus, indicates that the surface molecules of some nitro¬ methane solutions are more ordered than the surfaces of either of the components. A possible explanation for the greater ordering would be the formation of a complex between CC14 and CH3N02, a possibility dia¬ metrically opposed to the sugges¬

tions of de Maine et al. (1957) that a monomer-dimer nitroparaffin equi¬ librium occurs in these binary solu¬ tions. More likely, this effect is the demonstration previously unreport¬ ed, for surface layers of the entropy of mixing effect, previously demon¬ strated for the bulk solutions of CC14 and CH3N02 (Wettaw, et al. (1969)).

The surface entropies and, enthal¬ pies of mixing were calculated for each solution by taking the differ-

62

Transactions Illinois Academy of Science

ence between a value in Table 3 and its corresponding value predicted for perfect solutions based upon addi¬ tivity of components with respect to mole fraction. That is, the per¬ fect solution value is found by re¬ placing and M2 in Equation (3) with the appropriate thermodynamic values, e.g. (sm)i and (sm)2 to ob¬ tain smM. Molar excess surface entro¬ pies of mixing, s,„E, were calculated

bv addition of the terms &

i = 2

R 2 Xj In Xi

i = l

to the molar surface entropies of mixing, smM. Scratchard (1949) has shown, for bulk solutions, that excess volume of mixing, VE, can be fitted to a function of the form

VE = a0x1x2 + a1x1x2(x1 —

X2)+ . (4)

where a0 and ax are constants, and

Table 4.— Surface Mixing and Excess Thermodynamic Quantities.

Enthalpy

Entropy

M

M

Mole

M

S(T

M E

ho-

(ergs/cm2)

hm x 10 10 (ergs/ mole)

Fraction

(RN02)

(ergs/

cm2-deg)

sm x 10-7 smxl0-7 (ergs/ mole-deg)

30°C „

35°C

45 °C

30 °C

35°C

45 CC

Nitromethane

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.1

0.0001

0.52

3.2

—0.58

—0.54

—0.57

0.11

0.12

0.11

0.2

—0.0038

0.04

4.0

—2.5

—2.6

—2.5

—0.08

—0.17

—0.08

0.3

—0.021

—2.7

2.3

—8.3

—8.2

—8.3

—1.0

—1.0

—1.0

0.4

—0.021

—2.6

3.0

—8.9

—8.9

—9.0

—1.0

—1.0

—1.0

0.5

—0.046

—7.1

—1.3

—18.

—18.

—18.

—2.5

—2.5

—2.5

0.6

—0.053

—8.3

—2.7

—20.

—20.

—20.

—3.0

—2.9

—3.0

0.7

—0.053

—8.4

—3.3

—21.

—20.

—21.

—3.0

—3.0

—3.0

0.8

—0.044

—7.1

—3.1

—18.

—17.

—17.

—2.6

—2.5

—2.6

0.9

—0.035

—5.5

—2.8

—14.

—13.

—13.

—2.0

—1.9

—1.9

1.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

Nitroethane

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.1

—0.010

—1.9

0.80

—3.3

—3.4

—3.4

—0.59

—0.63

—0.60

0.2

—0.0052

—1.0

3.0

—2.3

—2.1

—2.3

—0.38

—0.38

—0.38

0.3

0.0037

0.60

5.7

0.41

0.45

0.41

0.10

0.098

0.10

0.4

—0.0074

—1.6

4.0

—3.3

—3.3

—3.3

—0.58

—0.63

—0.59

0.5

—0.0055

—1.3

4.5

—2.8

—2.8

—2.8

—0.52

—0.56

—0.52

0.6

0.0034

0.40

6.0

—0.18

0.0

—0.18

0.012

0.016

0.012

0.7

0.012

2.1

7.2

2.9

3.0

2.9

0.54

0.54

0.55

0.8

0.012

1.9

5.9

3.0

3.0

3.0

0.54

0.53

0.54

0.9

0.0041

0.70

3.4

0.73

0.75

0.73

0.19

0.18

0.19

1.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

Gunter et al — Nitroparaffin Surface Tensions

63

x4 and x2 are the mole fraction of CC14 and nitroparaffin, respectively. For binary solutions, the first term on the right hand side of Equation (4) is a quadratic term in either xx or x2 while the second term is a cubic term. Other molar excess quan¬ tities are expected, for bulk solution, to have the same functional form as Equation (4). Gunter et al. (1967) have shown for the bulk solutions of CH3N02— CC14 that only a0 is non¬ zero while for bulk solutions of C2H5N02— CC14 only ax is non-zero. The surface enthalpies of mixing,

which are identical with the excess surface enthalpies of mixing, shown in Table 4 indicate exactly the same functional information. That is, the cubic functional forms of the nitro- etliane values represent less devia¬ tion from ideality than the quad¬ ratic functional form of the nitro- methane values. The first example known to the present authors of the usual treatment of perfect solutions and excess functions to surface lay¬ ers rather than to bulk solutions is given by Bloom, et a?. (1960). Their application to molten salts is rather

Table 5. — Parachors of Carbon Tetrachloride-Nitroparaffin Solutions.

Calculated from Solution Surface Tensions

Calculated from Component Surface Tensions

Calculated from Atomic and Bond

Mole Fraction

(RN02)

30°C

35°C

-

45 °C

30°C

35°C

45 °C

—

Parachor

Values

Nitromethane

0.0 .

222.0

221.3

221.8

222.0

221.3

221.8

229.8

.1 .

212.7

211.8

212.2

213.1

212.4

212.9

220.1

.2 .

203.4

202.5

202.9

204.3

203.6

204.0

210.3

.3 .

193.7

193.4

194.0

195.4

194.7

195.1

200.6

.4 .

184.5

184.0

184.9

186.5

185.8

186.3

190.9

.5 .

174.7

174.2

176.2

177.7

177.0

177.4

181.2

.6 .

165.2

165.2

167.2

168.8

168.1

168.5

171.4

.7 .

156.2

156.3

158.4

159.9

159.2

159.6

161.7

.8 .

147.9

148.3

149.9

151.1

150.4

150.8

152.0

.9 .

140.1

140.5

141.5

142.2

141.5

141.9

142.2

1.0 .

133.3

132.7

133.0

133.3

132.7

133.0

132.5

Nitroethane

0.0 .

222.0

221.3

221.8

222.0

221.3

221.8

229.8

.1 .

216.2

215.9

216.6

216.9

216.2

216.7

224.1

.2 .

210.8

210.6

211.0

211.8

211.2

211.7

218.3

.3 .

205.8

205.4

205.7

206.6

206.2

206.7

212.6

.4 .

200.2

198.6

200.6

201.5

201.2

201.6

206.9

.5 .

195.0

193.2

195.4

196.4

196.1

196.6

201.2

.6 .

189.8

190.0

190.1

191.2

191.1

191.6

195.4

.7 .

185.1

185.2

185.1

186.1

186.1

186.6

189.7

.8 .

180.4

180.3

180.5

181.0

181.0

181.5

184.0

.9 .

175.6

175.4

175.9

175.9

176.0

176.5

178.2

1.0 .

170.7

171.0

171.5

170.7

171.0

171.5

172.5

64

Transactions Illinois Academy of Science

remote from the type of solutions discussed in this paper. The only other example of this treatment to surface layers known to the authors was published by Suri and Rama- krishna (1969) after the conclusion of the present work. However, the agreement with the conclusions of Gunter, et al. (1967) is a validation of the application to surface layers rather than an assumption that sur¬ face layers obey the same physical laws as the bulk solution.

The negative smE nitromethane values indicate (Scatchard and Ray¬ mond (1938))that some CC14 mole¬ cules are associating with the clusters of nitromethane molecules, thus, sub¬ stantiating the conclusion of ordered surfaces derived from the sa and sm values. The positive smE values indicate dissociation of the nitro- paraffins. In one sense, both of the conelusioss suggested by Table 3 are valid, if a complex is interpreted as a simple association of molecules. Reid and Sherwood (1966) suggest the use of the parachor for the esti¬ mation of surface tensions of non- aqueous mixtures. Table 5 compares the values of the parachor calculated directly from solution densities and surface tensions and those calculated from the parachors of the mixture components. The maximum devia¬ tions of 2.3% (CH3N02) and 1.5% (C2H5N02) from the linear mixture law indicate that estimation from parachor values is appropriate for these solutions. The greater devia¬ tions for the nitromethane solutions is attributed to the greater differ¬ ence in surface tension of the com¬ ponents as first observed by Ham- mick and Andrew (1929). The dis¬ crepancy between the observed CC14 parachor value and the value ob¬ tained by addition of atomic para¬ chors is attributed to the accumula¬ tion of negative groups (Mumford

and Phillips (1929)) and suggests that a better estimation of surface tensions of carbon tetrachloride - nitroparaffin binary solutions would result from the use of an experi¬ mental CC14 parachor.

Acknowledgment

The authors gratefully acknowledge the use of the Data Processing and Computing Center of Southern Illinois University. They also wish to thank Commercial Solvents Corporation for complimentary samples which were used in the initial phases of this research. The authors are greatly indebted to Professor K. A. Van Lente for his assistance with experimental procedures. This work was supported by a grant from the Petroleum Research Fund (602-B) administered by the American Chemical Society.

Literature Cited

Bloom, H., F. G. Davis, and D. W. James. 1960. Molten Salt Mixtures. Part 4. The Surface Tension and Surface Heat Content of Molten Salts and Their Mix¬ tures. Trans. Faraday Soc. 56; 1179- 1186.

Boyd, G. E. and C. E. Copeland. 1942. Surface Tensions, Densities, and Para¬ chors of the Aliphatic NitroparafFins. J. Am. Chem. Soc. 64: 2540-2543.

Daniels, F., J. W. Williams, P. Bender, R. A. Alberty, and C. D. Cornwell. 1962. Experimental Physical Chemistry. 6th Ed. p. 401-402, McGraw-Hill, New York.

Defay, R., I. Prigogine, and A. Belle- mans, 1966. Surface Tension and Ad¬ sorption. Trans, by D. H. Everett, p. 92-93, John Wiley & Sons, Inc., New York, XIII x 432 pp.

Demaine, P. A. D., M. M. Demaine, and A. G. Goble, 1957. Experimental Stud¬ ies of Solution Processes. Part 1. Evi¬ dence of the Dimerization of Nitrome¬ thane in Carbon Tetrachloride and in Cyclohexane. Trans. Faraday Soc. 53: 427-432.

Guggenheim, E. A., and N. K. Adam. 1933. The Thermodynamics of Ad¬ sorption at the Surface of Solutions. Proc. Roy. Soc. A139: 218-236.

Gunter, C. R., J. F. Wettaw, J. D. Drennan, R. L. Motley, M. L. Coale, T. E. Hanson, and B. Musulin, 1967. Densities and Molar Volumes of Binary

Gunter et al — Nitroparaffin Surface Tensions

Table 6. — Summary of Literature Values.

65

Entropy

Surface

Tension

Enthalpy

30°C, y

S<x

sm x 10 7

ho-

Parachor

(dyne/cm)

(ergs/cm2-deg)

(6rgs/ mole-deg)

(ergs/cm1 2)

[P]

Nitromethane

34. 262

0.13877

31.597

81. 14

131. 814

35. 473

0.1504

15. 85

86. 43

132.63-8

35. 484

0.16783

14.48

81. 37

132. 77

35. 501

0.1605

20. 33

84. 45

35.9s

0.14648

79. 88

32. ID2

Nitroethane

31. 53

0.10907

12. 837

69. 83

171.07"8

0.12553

14. 19

65. 79

171. 13

0.11639

14. 98

67. 17

171. 215

0.12188

18. 43

67. 98

Carbon Tetrachloride

25. 546

0.1194

17. 612

61. 74

219. 316

25.5 74

0.125910

17. 811

63. 610

219. 617

0.13278

18. 16

66. 58

220. O’5

*45 . 1 °C

0.117311

18. 713

20. 38

60. 511

221. 08

1. Suri and Ramakrishna (1969)

4. Hennaut-Roland and Lek (1931)

7. Boyd and Copeland (1942)

10. Pugachevich et al. (1963)

13. Morino (1932)

16. Ray (1934)

2. Morgan and Stone (1913)

5. Thompson et al. (1954)

8. Vogel (1948)

Solutions of Nitroparaffins in Carbor Tetrachloride. J. Chem. Eng. Data 12: 472-474.

Hammick, D. L. and L. W. Andrew. 1929. The Determination of the Para- chors of Substances in Solution. J. Chem. Soc. 1929 : 754-759.

Harkins, W. D. and Y. C. Cheng. 1921. The Orientation of Molecules in Sur-

11. Ramsay and Aston (1894)

14. Hammick and Andrew (1929) 17. Mumford and Phillips (1950)

3. Snead and Clever (1962)

6. Harkins and Cheng (1922)

9. Ramsay and Shields (1893)

12. Renard and Guye (1907)

15. Mumford and Phillips (1929)

faces. VI. Cohesion, Adhesion, Tensile Strength, Tensile Energy, Negative Sur¬ face Energy, Interfacial Tension, and Molecular Attraction. J. Am. Chem. Soc. 43: 35-53.

Hennaut-Roland, Mme. and M. Lek, 1931. Methods and Equipment Used at the Bureau of Physico-Chemical Standards. IV. The Surface Tension

66

Transactions Illinois Academy of Science

of a Series of Organic Substances. Bull. Soc. Chem. Belg. 40: 177-94.

Morgan, J. L. R. and E. C. Stone. 1913. The Weight of a Falling Drop and the Laws of Tate. XII. The Drop Weights of Certain Organic Liquids and the Surface Tensions and Capillary Con¬ stants Calculated from Them. J. Am. Chem. Soc. 35: 1505-1523.

Morino, Y. 1932. The Surface Tensions of Binary Mixtures. Bull. Inst. Phys. Chem. Research (Tokyo) 11: 1018-

1043.

Mumford, S. A. and J. W. C. Phillips. 1929. The Evaluation and Interpreta¬ tion of Parachors, J. Chem. Soc. 1929: 2112-2133.

Mumford, S. A. and J. W. C. Phillips. 1950. The Physical Properties of Some Aliphatic Compounds. J. Chem. Soc. 1950: 75-84.

Partington, J. R. 1951. “An Advanced Treatise on Physical Chemistry”. Vol. 2. “The Properties of Liquids”. Long¬ mans, Green & Co., London, xliv + 448

pp.

PUGACHEVICH, P. P., L. A. NiSEL'sON, T. D. Sokolova, and N. S. Anurov. 1963. Density, Viscosity, and Surface Tension of Carbon and Stannic Tetra¬ chlorides. Zhur. Neorgan. Khim. 8: 791-6, cf. C. A. 1964. 60: 7483e. Ramsay, W. and E. Aston. 1894. Die molekulare Oberflachenergie von Mis- chungen sich nicht associierender Flus- sigkeiten. Z. f. Phys. Chem. 15: 89-97.

Ramsay, W. and J. Shields. 1893. Uber die Molekulargewichte der Fliissigkeiten. Z. f. Phys. Chem. 12: 433-475.

Ray, S. K. 1934. Determination of Parachor in Solution. Part I. J. Ind. Chem. Soc. 11: 671-679.

Reid, R. C., and T. K. Sherwood, 1966. The Properties of Gases and Liquids. 2nd Ed. p. 384-385, McGraw-Hill, New York. xx + 646 pp.

Renard, Th. and Ph. — A. Guye. 1907. Mesures de Tensions Superficielles a

L’air Libre. J. chim. phys. 5: 81-112.

Scatchard, G. and C. L. Raymond, 1938. Vapor-Liquid Equilibrium. II. Chloro¬ form-Ethanol Mixtures at 35, 45, and 55°. J. Am. Chem. Soc. 60: 1278-

1287. ^

Snead, C. C. and H. L. Clever. 1962. Thermodynamics of Liquid Surfaces. Surface Tensions of Eight High Purity Nitroparaffins from 0° to 60° C. J. Chem. Eng. Data 7 : 393-394.

Suri, S. K. and V. Ramakrishna. 1969. Surface Tension of Binary Solutions of Nitromethane in Benzene and Dioxane. Ind. J. Chem. 7: 243-247.

Thompson, C. J., H. J. Coleman, and R. V. Helm. 1954. The Purification and Some Physical Properties of Nitro¬ methane. J. Am. Chem. Soc. 76: 3445- 3446.

Timmermans, Jean. 1959. The Physico- Chemical Constants of Binary Systems in Concentrated Solutions. Vol. 1. Two Organic Compounds (without Hydroxyl Derivatives). Interscience Publ. Inc., N.Y. xiii + 1259 pp.

Van Rysselberghe, P. 1938. Conven¬ tions and Assumptions in the Interpreta¬ tion of Experimental Data by Means of the Gibbs Adsorption Theorem. J. Phys. Chem. 42: 1021-1029. _

Vogel, A. I. 1948. Physical Properties and Chemical Constitution. Part XXIII. Miscellaneous Compounds. Investigation of the So-called Co-ordinate or Dative Link in Esters of Oxy-acids and in Nitro- paraffins by Molecular Refractivity De¬ terminations. Atomic Structural and Group Parachors and Refractivities. J. Chem. Soc. 1948: 1833-1855.

Wettaw, J. F., E. E. McEnary, J. D. Drennan, and B. Musulin. 1969. Viscosities of Binary Solutions of Nitro¬ paraffins in Carbon Tetrochloride. J. Chem. Eng. Data 14: 181-184.

Worthing, A. G. and J. Geffner, 1943. Treatment of Experimental Data. J. Wiley, New York, pp. 170-171. Manuscript received August 5, 1970

COMPUTERIZED CURVE FITTING:

AN ALTERNATIVE TO GRAPHICAL INTERPRETATION

BORIS MUSULIN

Southern Illinois University, Carbondale, Illinois

Abstract. — A comparative study is made of the use of statistical techniques in determining the degree of an empirical polynomial in a problem with experimental error. Each problem requires information on error limits and the magnitude of the dependent and independent variables for establishing a statistical criterion. Data for densities of binary solutions of nitro- methane or nitroethane in carbon tetra¬ chloride are used for examples. Concen¬ tration, temperature, and excess functions are examined.

The purpose of this paper is to investigate criteria, determining the degree of an empirical polynomial, which appeal to chemists (as well as other scientists whose data contains experimental error). Particular em¬ phasis is placed upon first degree (linear) functions because of their common occurrence in chemical prob¬ lems.

The availability of high speed com¬ puters suggests the feasibility of an alternate to the “eye-ball” graphi¬ cal techniques long used by chemists in analysis of laboratory data. Gra¬ phical analysis involving vagaries such as choice of scale, artistic care, etc., is as much an art as a science while a computer alternative removes the subjective element from the an¬ alysis. An ideal criterion has two attributes, viz it can be routinely programmed for use on a computer and it is based upon concepts famil¬ iar to chemists. Proposed criteria are related to formalized statistical techniques.

The various techniques are applied to density data of binary mixtures of nitromethane and nitroethane which were reported by Gunter, et. al. (1967). These data are neither complete enough nor accurate enough to warrant extensive research treat¬ ment but they are sufficient for peda¬ gogical purposes. Consequently, the reader is cautioned that the chemical conclusions are only indicative, not definitive.

The Quintic Concentration Equation

For each temperature reported by Gunter, et. al. (1967), the best co¬ efficients, ai? for density as a func¬ tion of concentration

i = 5

cl = ^j^aiC1 (1)

i = 0

were determined by a least squares procedure (Daniels, et. al., 1962) contained as part of a standard re¬ gression program (Purcell, 1965) available at the Data Processing and Computing Center of Southern Illi¬ nois University at Carbondale, (Ni¬ tromethane at 45 °C was not included in this experiment due to insufficient data). Three different representa¬ tions of concentration were used, x2, weight fraction of nitroparaffin, viz. mole fraction of nitroparaffin,

[67]

68

Transactions Illinois Academy of Science

w2, and volume fraction of nitro- paraffin, v2. In each case, the nitro- paraffin was assumed to be a monom¬ er. These calculations were per¬ formed on an IBM 7044 computer using FORTRAN IV as the com¬ puter language.

The choice of a quintic degree was arbitrary but with the expectation that the degree was sufficiently high as to contain terms of no statistical significance. The coefficients are giv¬ en in Table 1. The value of the mix¬ ture density at zero concentration is the carbon tetrachloride density at the specified temperature, irre¬ spective of the concentration repre¬ sentation or the nature of the second solution component. The a0 values are in close agreement with each

other as well as with experiment ex¬ cept for C2H5N02 at 35°C which agrees somewhat less well with ex¬ periment. The slightly poorer 35° C C2H5N02 a0 value is clearly due to the data, a conclusion which can be verified by an examination of the data and their errors reported by Gunter, et. al. (1967). In Table 1, the values of the mixture density calculated for unit concentration are also listed. These values are the nitroparaffin density values at the specified temperature. The excep¬ tional agreement between sets and between calculated and experimental values indicates that any faults lie with the data near zero concentra¬ tion. Every value reported in Table 1 has four or five significant figures

Table 1. — Calculated Constants for Quintic Concentration Equations.

Nitromethane

ai

a2

a3

a4

as

Temp.

a0

(g/ ml-

(g/ml-

(g/ ml-

(g/ml-

(g/ml-

d(conc=l)

(°C)

(g/ml)

cone)

cone2)

cone3)

cone4)

cone5)

(g/ml)

d VS. X2

30

35

45

1.5749

1.5662

—0.2624

—0.2448

—0.1400

—0.2820

0.04214

0.4054

—0.1049

—0.4713

0.007572

0.1378

1.1173

1.1113

d VS. W2

30

35

45

1.5750

1.5667

-0.6757

—0.6789

0.3752

0.3613

—0.3013

—0.1868

0.2139

0.04097

—0.06987

0.007873

1.1172

1.1112

d Vi. V2

30

35

45

1.5749

1.5665

-0.4767

—0.4684

0.02663

-0.07702

0.007180

0.3432

—0.03319

—0.4280

0.01837

0.1751

1.1172

1.1114

Musulin — Computerized Curve Fitting

69

Nitroethane

ai

a2

a3

a4

as

Temp.

aG

(g/ml-

(g/ ml-

(g/ ml-

(g/ml-

(g/ ml-

d(conc=l)

(°C)

(g/ml)

cone)

cone2)

cone3)

cone4)

cone5)

(g/ml)

d VS. X2

30

1.5748

—0.4000

—0.1681

0.2476

—0.3825

0.1628

1.0346

35

1.5670

—0.5201

0.8910

—2.5015

2.4683

—0.8753

1.0294

45

1.5455

—0.4251

0.09944

—0.4724

0.4413

—0.1719

1.0168

d VS. W2

30

1.5749

—0.8377

0.5220

—0.3867

0.1983

—0.03624

1.0346

35

1.5660

—0.8859

1.3898

—3.4098

4.0286

—1.6599

1.0288

45

1.5452

— 0 . 8286

0.6076

—0.6634

0.5452

—0.1892

1.0168

d VS. V2

30

1.5749

—0.5471

— 0.004345

0.1398

—0.2630

0.1344

1.0347

35

1.5673

—0.6922

1.4641

—4.1008

4.5409

— 1 . 7504

1.0289

45

1.5454

—0.5618

0.2168

—0.4943

0.5024

—0.1916

1.0169

depending’ upon whether the abso¬ lute value is less or greater than unity. Gunter, et. al. (1967) indi¬ cate an experimental uncertainty in x2 of 5 x 10'3, which, for 30° C CH3N02, leads to

d = d± 10~3(1.31 + 1.40 x —

0.63 x22 + 2.10 x32 — 0.189x42) (2)

where the expression following the ± is the error due to the error in x2. For x2 = 0, an error of 0.001 re¬ sults and for x2 = l, an error of 0.004 results. The conclusion is that the number of significant figures in Table 1 are useful only to prevent rounding errors during calculation and that all final calculated densi¬ ties should be rounded to four sig¬ nificant figures. From this view¬

point, it is clear that at each tem¬ perature all a0 values are in perfect agreement. In other words, the ex¬ perimental scientist would not con¬ sider the 35° C C2H5N02 value as slightly poorer but merely a devia¬ tion within the range of experiment¬ al error. Since, as stated in the in¬ troduction a criterion for chemists is sought, the latter viewpoint must prevail.

Many chemists have made little use of statistical tools excepting means and standard deviations. The standard program (Purcell, 1965) produced the variance after the ad¬ dition of each power of the inde¬ pendent variable. The familiar standard deviation was derived b}r taking the square root of the vari¬ ance divided by the degrees of free-

70

Transactions Illinois Academy of Science

dom (number of observations less the number of constants determined by least squares). These results are pre¬ sented in Table 2. In both the x2 and w2 representations, an error results in the second decimal place if only the linear term is used thereby in¬ dicating that the linear term is in¬ sufficient. For 30° C CH3NOo the use of x22 gives a standard deviation equal to the maximum error due to experimental error in the mole frac¬ tion while x32 gives a standard de¬ viation equal to the minimum error. Depending upon the experimenter, either the quadratic or cubic term is sufficient. In either case, it is clear that an individual error analysis is required for each compound, at each temperature. In the present re¬ search, the decision to accept the maximum error due to experiment was made. The data in Table 2 in¬ dicates that the quadratic term is sufficient in the mole fraction and weight fraction representations and that the linear term is sufficient in the volume fraction representation. Figures 1 through 3 graphically de¬ pict the density as a function of vol¬ ume fraction. Every chemist would agree that both sets of nitroparaffin data are well represented by a linear function thereby verifying the suf-

Figure 1. Density of Nitroparaffinic Binary Solutions as a Function of Volume Fraction of Nitroparaffins at 30 °C.

Figure 2. Density of Nitroparaffinic Binary Solutions as a Function of Volume Fraction of Nitroparaffins at 35°C.

Figure 3. Density of Nitroparaffinic Binary Solutions as a Function of Volume Fraction of Nitroparaffins at 45 °C.

ficiency of linear terms. The fact that density is a linear function of concentration, expressed in volume units, indicates the solutions are ideal ( Weissberger, 1959).

This linearity also substantiates the conclusions of MacFarlane and Wright (1933) that volume fraction is the most suitable independent variable in a density function.

The per cent deviations suggested by Foley, et. al. (1964) were cal¬ culated by dividing each entry of Table 2 by the means of all observa¬ tions of that nitroparaffin at the tem¬ perature of that entry. The results

Table 2. — The Standard Deviation:

Musulin — Computerized Curve Fitting

71

in

x

O VO

on Os lo lo oo l o

O co O o o o o o o

o r-

LO'OrH

LON N

O co O

o o o o o o

CO N~

LO LO O

lo Ov vo

O co O o o o o o o

d d d

odd

dod

X

oo

CONrt

LO OV VO

O CO O

o o o

O O O

VO -H

O >— 1 NO i_o 1-0

O O

o o o o o o

ON CO

N- o N"

lo O VO

O LO o o o o o o o

Nitroethane

odd

odd

odd

co

X

0.000568

0.00427

0.000580

0.000925

0.00417

0.000921

0.000750

0.00488

0.000658

X

on co

N- O T-H i on

O O O

O O O

VO VO oo

NOvO

Nforo

O O O

O O O

On

CO LO ON

o- o

O "f 1“H

O O O

O O O

odd

odd

odd

X

OOc^N

LO OO LO

i— H r-H t— H

o o o

O — I-1 i NOh (N(N(N

O O O

VO

ciOON

o- —t o

O LO i-H

o o o o o o

odd

dod

odd

m

X

NO i— i •

o-v O

OO VO •

o o • o o • o o •

LO Ov

ON VO •

OO •

o o • o o • o o •

O ON •

O LO

ON N» •

O O ■

O O •

O O •

<N

X

d o •

<N

>

d o •

<N

h>

d d •

■'■f1 o- VO l-H •

o- vo •

o o •

O O •

O O ■

d vs.

o —i •

VO o •

N- N- •

o o • o o • o o •

*— H N-

CO VO

OO O'- •

o o ■ o o • o o •

4>

o o •

d d •

d d •

Nitromethan

CO

X

0.000967

0.00113

0.000775

0.000769

0.000774

0.000710

X

N- oo •

t-H ON .

o o • o o •

OO ON •

O oh

OH ON •

o o • o o •

LO ON

lo vo

O', oo •

o o •

o o •

o o •

d d •

d d •

O O ;

X

Tt^OOH

■rf co in rl in rN

O O O

vo oo o

VO VO Ov

i— H i— H <

o o o

-f •f vO lo Ov O

iHfHfO

o o o o o o

odd

odd

odd

nperature

(°C)

<D

H

O LO LO co co Tfi

o LO LO

CO CO Tt*

O LO LO

CO CO 'f

a All entries stated in units of g/ml.

72

Transactions Illinois Academy of Science

given in Table 3 show that the per cent deviations corresponding to terms judged of sufficient degree by the criterion of standard deviation are of 0(10'1%). Consequently, an error analysis, as in the case of standard deviation, is used to estab¬ lish a criterion for judging sufficien¬ cy of degree. Another observation is that the percent deviation (or the standard deviation) does not change, perceptibly, if terms of no statistical significance are added. For example, with 30° C CH3N02, cubic, quatric, and quintic terms in x2 give a per cent deviation of 0.06% indicating the cubic term is sufficient, but as noted, the use of experimental error determined the quadratic term is suf¬ ficient. The difference between the experimentalist and non-experiment¬ alist is further emphasized. In the case of 35 °C C2H5N02 the two view¬ points merge, indicating a quadratic term in x2 is sufficient since per cent deviation remains constant at 0.3%. the proper value considering exper¬ imental error. Of course, the fact that even the use of a quintic term does not reduce the per cent devia¬ tion again shows the difference in quality of this particular set of data.

The standard computer program also yielded the value of R2, the square of the multiple correlation co¬ efficient (Baten, 1938) after the ad¬ dition of each power of the inde¬ pendent variable. These values are given in Table 4. The values of R2 are also the sum of the proportions of variance through the term being added (Musulin and Musulin, 1967). Consequently a value of 1 indicates that all variance has been accounted for within the number of significant figures yielded by the standard pro¬ gram, i.e. five significant figures for R2. An examination of Table 4 in¬ dicates that the 35° C nitroethane data is generally not comparable to

the data at 30°C and 45°C, again emphasizing the experimental errors in 35°C data. Excepting this 35°C C2H5N02 data from a statistical viewpoint, the mole fraction repre¬ sentation recovers all the variance with terms through the cubic de¬ gree, as is true for the weight frac¬ tion representation with nitroethane. For the volume fraction representa¬ tion, quadratic terms are sufficient for nitromethane and linear terms for nitroethane. Once again, the view¬ point of the experimentalist must be introduced for the use of statistics alone results in equations of greater refinement than can be warranted by experimental error. Figures 1 through 3 show that the requirement of quadratic v2 terms suggested by statistical methods for the CH3N02 data is too stringent.

Foley, et. al. (1964) have suggest¬ ed that a criterion for linearity, within 6%, is |rj > 0.995. It is pro¬ posed to generalize this criterion for use with polynomials of any degree by establishing a lower bound for R2 (this particular standard pro¬ gram yields R2 but a lower bound on the absolute value of R would serve as well). The sum of squared deviations (and hence, the per cent deviation) is proportional to vari¬ ance of the dependent variable and to (1 -R2) (Baten, (1938)). Since the range of the dependent variable is essentially the same for each nitro- paraffin, the average criterion R2 > 0.998 can be established for deter¬ mining the proper polynomial de¬ gree to be used in an empirical equa¬ tion. This criterion provides the same results as were obtained with the standard deviation and the per cent deviation, i.e. quadratic equa¬ tions are appropriate with x2 and w2, and linear equations with v2.

An alternate statistical technique to determine degree of the inde-

Musulin — Computerized Curve Fitting

73

Table 3. — The Per Cent Deviation.

Nitromethane

Nitroethane

Temp.

(°C)

X

X2

X3

X4

X5

X

X2

X3

X4

X6

d vs. X2

30

1.76

0.301

0.0698

0.0551

0.0603

1.19

0.129

0.0427

0.0437

0.0391

35

1.73

0.311

0.0821

0.0448

0.0437

1.38

0.307

0.323

0.300

0.294

45

2.57

1.20

0.163

0.0444

0.0474

0.0426

d VS. W2

30

1.20

0.150

0.0559

0.0549

0.0596

1.66

0.208

0.0696

0.0381

0.0414

35

1.22

0.176

0.0559

0.0510

0.0559

1.54

0.299

0.315

0.341

0.283

45

1.39

1.62

0.236

0.0706

0.0562

0.0549

d VS. V2

30

0.111

0.0545

0.0559

0.0600

0.0656

0.0554

0.0556

0.0564

0.0430

0.0416

35

0.141

0.0627

0.0516

0.0558

0.0552

0.391

0.359

0.369

0.378

0.298

45

0.224

0.0820

0.0782

0.0504

0.0516

0.0465

Vm vs. x2

30

0.0975

0.0474

0.0505

0.0546

0.0593

0.0424

0.0450

0.0410

0.0366

0.0303

35

0.127

0.0514

0.0542

0.0527

0.0482

0.333

0.301

0.319

0.294

0.286

45

0.220

0.0691

0.0714

0.0447

0.0462

0.0410

Vm vs. w2

30

4.96

1.24

0.294

0.0831

0.0610

2.06

0.406

0.0484

0.0312

0.0328

35

4.93

1.21

0.290

0.0951

0.0686

2.23

0.621

0.315

0.322

0.272

45

6.16

2.10

0.378

0.0703

0.0498

0.0516

Vm vs. v2

30

3.12

0.490

0.0855

0.0541

0.0577

0.866

0.0616

0.0361

0.0302

0.0288

35

3.09

0.472

0.0876

0.0561

0.0549

1.04

0.298

0.274

0.280

0.226

45

4.29

0.890

0.0539

0.0470

0.0482

0.0472

74

Transactions Illinois Academy of Science

Table 4. — The Multiple Correlation Coeffiecient.

Temp.

(°C)

Nitromethane

Nitroethane

X

X2

X3

X4

X5

X

X2

X3

X4

X5

d VS. X2

30

35

45

0.9763

0.9771

0.9890

0.9994

0.9993

1.000

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

0.9930

0.9905

0.9927

0.9999

0.9996

0.9999

1.0000

0.9996

1.0000

1.0000

0.9997

1.0000

1.0000

0.9998

1.0000

d VS. W2

30

0.9890

0.9998

1.0000

1.0000

1.0000

0.9864

0.9998

1.0000

1.0000

1.0000

35

0.9886

0.9998

1.0000

1.0000

1.0000

0.9882

0.9996

0.9996

0.9996

0.9998

45

0.9968

1.0000

0.9868

0.9997

1.0000

1.0000

1.0000

d VS. V2

30

0.9999

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

35

0.9998

1.0000

1.0000

1.0000

1.0000

0.9992

0.9994

0.9995

0.9995

0.9998

45

0.9999

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

Vh vs. x2

30

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

35

1.0000

1.0000

1.0000

1.0000

1.0000

0.9990

0.9993

0.9993

0.9995

0.9996

45

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

1.0000

Vm vs. w2

30

0.9371

0.9965

0.9998

1.0000

1.0000

0.9600

0.9986

1.0000

1.0000

1.0000

35

0.9377

0.9966

0.9998

1.0000

1.0000

0.9534

0.9968

0.9993

0.9994

0.9996

45

0.9758

1.0000

0.9594

0.9988

1.0000

1.0000

1.0000

Vm vs. v2

30

0.9750

0.9994

1.0000

1.0000

1.0000

0.9930

1.0000

1.0000

1.0000

1.0000

35

0.9753

0.9995

1.0000

1.0000

1.0000

0.9899

0.9993

0.9995

0.9995

0.9997

45

0.9883

1.0000

0.9927

1.0000

1.0000

1.0000

1.0000

Musulin — Computerized Curve Fitting

75

pendent variable which is required in the empirical equation is the Analysis of Variance (Bennett and Franklin (1954)). In order to con¬ firm the statistical conclusions drawn from Table 2, an analysis of vari¬ ance was performed on the 30 °C CH3N02 data. At the 1% level, the F-ratio test indicated x32, w22, and v22 were significant. This informa¬ tion is identical to that derived from Table 4. Statistically the analysis of variance is equivalent to the use of R2 made in this study but inser¬ tion of the experimentalists view¬ point would require redefining F- ratio test values at the various levels of significance.

In an ideal mixture, the molar vol¬ ume, VM, is a linear function of the mole fraction of the solute, x2, (Rowlinson, 1959). The plots of VM vs. x2; Figures 4 through 6, are linear. In order to verify the valid¬ ity of the criteria which have been suggested, the experiment of substi¬ tuting VM for the dependent varia¬ ble in Equation (1) was performed. The per cent deviation and the values of R2 are given in Tables 3 and 4, respectively. An error analvsis for 30°C CH3N02 indicates 0.39% > per cent deviation > 0.23% for 0= x2= 1- Thus the maximum per

Figure 4. Molar Volume of Nitroparaf- finic Binary Solutions as a Function of Mole Fraction of Nitroparaffins at 30°C.

v«

Figure 5. Molar Volume of Nitroparaf- finic Binary Solutions as a Function of Mole Fraction of Nitroparaffins at 35°C.

Vm

Figure 6. Molar Volume of Nitroparaf- finic Binary Solutions as a Function of Mole Fraction of Nitroparaffins at 45°C.

cent deviation is approximately the same as the maximum per cent de¬ viation for density even though the values of VM are approximately 50 times greater than the values of d reaffirming the contention of Foley, et. al. (1964) with regards to per cent deviation.

From the per cent deviation, lin¬ ear equations are appropriate if x2 is the independent variable and quadratic equations are appropriate (barely so for CH3N02) if v2 is the independent variable. With w2 as the independent variable, the per cent deviation indicates quadratic equations with nitroethane and cubic

76

Transactions Illinois Academy of Science

equations with nitromethane. As usual, 35° C C2H5N02 is slightly poorer than the other data. For this choice of dependent variable the range of the dependent variable is rather different for the two nitro- paraffins resulting in a standard de¬ viation for nitromethane 1% times larger than the standard deviation of nitroethane. The difference in standard deviation leads to two dif¬ ferent exact criteria, viz. R2 > 0.999 (CH3N02) and R2 > 0.998 (C2H502). These separate criteria re¬ produce exactly the information giv¬ en by per cent deviation.

An exact analysis of variance for 30 °C nitromethane indicates that quadratic terms in the x2 representa¬ tion, quatric terms in the w2 repre¬ sentation, and cubic terms in the v2 representation are required. As with the density data, terms of degree one higher are required by the use of pure statistics than are required by an experimentalist desiring correct¬ ness within the experimental error defined by Gunter, et. al. (1967).

Final Concentration Equations

The least squares procedure was repeated with the density and molar volume data, the degree of each em¬ pirical polynomial being that degree determined by the criteria establish¬ ed in this work. The coefficients which result are slightly different than those presented in Table 1 since the minimization is accomplished with less variables. The new coeffi¬ cients are given in Table 5. The values of the standard deviation, per cent deviation, and R2 given in Ta¬ bles 2, 3, and 4 for each degree of freedom are invariant and desired information of this type may be read directly from the appropriate col¬

umn of the suitable table. Entries have been made in Tables 2, 3, and 4 from the quadratic and linear fits of the three point 45° C CH3N02. The R2 value does not have the same significance as adjudged by the Stu¬ dent t-test (Baten (1938) ) . Of course, the resulting minimization yields predicted values which are not as good, mathematically, as those ob¬ tained by the quintic equations but the range of a0 values and the values of the functions at unit concentra¬ tion (also listed in Table 5) are within the appropriate tolerance ranges.

In those cases where quadratic co¬ efficients are listed, the value of a2 indicates the amount of curvature, i.e. the lack of linearity. Further, extrema of the quadratic functions lie outside the region of physical sig¬ nificance, i.e. 0 ^ cone. 5j=l. For density as a function of v2, the sec¬ ond derivative of a quadratic fit, in every case, is positive which indi¬ cates the curvature of each plot is convex downward. It should be noted that the magnitude of the sec¬ ond derivative also shows that the amount of such curvature is very small. In such plots, Usol’tseva (1960) attributes a curvature which is convex downward to an associ¬ ated compound dissociating into an¬ other component. Usol’tseva’s con¬ clusion is in accord with the concept of a nitroparaffin dimer dissociat¬ ing, slightly, into a monomer.

Temperature Functions

For each mole fraction of each solution, the density was fitted, by a least squares procedure, to a lin¬ ear function of temperature (Equa¬ tion (1) with the temperature, t, as the independent variable and the up¬ per limit i = 1). The linear form was chosen for two reasons ; the tern-

Musulin — Computerized Curve Fitting

Table 5. — Calculated Constants for Final Concentration Equations.

77

Temperature

(°C)

Nitromethane

Nitroethane

3o

(g/ml)

-ai

(g/ml-

conc)

a2

(g/ ml- conc2)

d(Conc

=1)

&0

(g/ml)

-ai

(g/ ml- conc)

a2

(g/ ml-

cone2)

Per Cent d(conc -1)

d VS. X2

25a

1.5796

0.1977

—0.2447

1.1372

30

1.5702

0.2009

—0.2462

1.1231

1.5727

0.3755

—0.1608

1.0364

35

1.5619

0.2042

—0.2404

1.1173

1.5629

0.3504

—0.1829

1.0296

45

1.5456

0.2378

—0.2101

1.0977

1.5426

0.3637

—0.1592

1.0197

“Brown & Smith (1955)

d VS. W2

30

1.5723

0.6217

0.1686

1.1192

1.5709

0.7572

0.2236

1.0373

35

1.5636

0.6207

0.1706

1.1135

1.5628

0.7378

0.2053

1.0303

45

1.5456

0.6504

0.2025

1.0977

1.5407

0.7346

0.2140

1.0201

d vs. V2

30

35

45

1.5728

1.5637

1.5437

0.4577

0.4549

0.4465

1.1151

1.1088

1.0972

1.5746 1.5687 1 . 5446

0.5398

0.5370

0.5259

1.0348

1.0317

1.0187

Temp.

(°C)

Nitromethane

Nitroethane

3.0

(ml/g)

-ai

(ml/ g- conc)

a2

(ml/ g- conc2)

-a3

(ml/ g- conc3)

VM

(Cone

=1)

3o

(ml/g)

-ai

(ml/ g- conc)

a2

(ml/g-

conc2)

VM

(Cone

=1)

Vm vs. x2

30

97.776

43.049

54.727

97.697

25.175

72.522

35

98.332

43 . 290

55.042

98.097

25.327

72 . 770

45

99.625

43.990

55.635

99.599

25 . 894

73.705

Vm vs. w2

30

35

45

97.395

97.911

99.527

96.548

96.533

96.850

87.748

86.927

52.916

34.071

33.482

54.524

54.824

55.593

97.210

97.712

99.117

42.034

43.236

43.393

17.734

18.909

18.413

72.910

73.385

74.137

Vm vs. v2

30

97.153

66.252

24.135

55.036

97.624

32.549

7.536

72.611

35

97.685

66.478

24.111

55.318

98.192

33.872

8.766

73.086

45

99.527

72.811

28.877

55.593

99.533

33.592

7.889

73.830

78

Transactions Illinois Academy of Science

perature range used was narrow and the maximum of three temperature points would allow for compensation of experimental error. The results are given in Table 6. The per cent deviation is also tabulated. Where only two temperature points were available, no error per cent is given. These and succeeding calculations were performed on an IBM 1620 computer with 40K storage using an IBM PR 025 monitor. The programs were written in FORTRAN II (IBM, 1962).

Except for 0.4 and 0.5 nitro- ethane, all per cent deviations are 0(10'2). This order of magnitude in¬ dicates a better fit than is warrant¬ ed by the data. It further substan¬ tiates the validity of choice of linear form. Although the exceptional data have per cent deviations appropri¬ ate to this study, these two sets are not of the same quality. Combining this information with the informa¬ tion from the concentration data, the two slightly poorer data points are 0.4 and 0.5 35°C C2H5N02. Various literature values are pre¬ sented in Table 8. Intercepts and

slopes calculated from several sets of density data in the literature have been included in Table 8.

Excess Functions

Scatchard (1949) has found that the excess molar volume of mixing, VE, can be fitted to a series,

(3)

i = n

VE = ^^^aix1x2(x1 -x2)‘ i = 0

where the ai are constants. The VE values calculated by Gunter, et. al. (1967) were fitted by a least squares procedure to a two-term equation of the form given in Equation (3) (i = 0 to 1) . Least squares fits were also made of single term equations (i = 0 and i = 1, corresponding to quadratic and cubic equations, re¬ spectively, in x2). A discard cri¬ terion (Worthing and Geffner, 1943) was applied in the fitting proced¬ ures. The coefficients for each case are summarized in Table 7.

The per cent deviation is also

Table 6. — Calculated Constants for Linear Temperature Equations.

Nitromethane

Nitroethane

Mole

Fraction

-ai x 103

Per Cent

a0

-ai x 103

Per Cent

(RN02)

(g/ ml)

(g/ml-°C)

Deviation

(g/ ml)

(g/ml-°C)

Deviation

0.0

1 . 6344

1.9686

0.03

1.6344

1.9686

0.03

0.1

1.5943

1.5740

1.5939

2.0073

0.03

0.2

1.5679

1.6940

1.5476

1.9096

0.05

0.3

1.5424

1.9333

0.02

1.4971

1.7567

0.06

0.4

1.4980

1.7180

1.4553

1.8293

0.37

0.5

1.4661

1.9180

1.4030

1.7731

0.41

0.6

1.4067

1.4580

1.3378

1.5503

0.01

0.7

1.3585

1.5220

1.2743

1.3794

0.01

0.8

1 . 2989

1.4160

1.2096

1.2877

0.03

0.9

1.2296

1 . 2620

1.1410

1.1883

0.09

1.0

1.1566

1.3066

0.02

1.0703

1.1841

0.01

Musulin — Computerized Curve Fitting

Table 7. — Calculated Constants for Volume of Mixing Equations.

79

Nitromethane

Nitroethane

Temp.

(°C)

a0

(ml/ mole- (mole fraction)2)

-ai

(ml/ mole- (mole fraction)3)

Error

Per Cent

a0

(ml/ mole- (mole fraction)2)

-ai

(ml/mole-

(mole

fraction)3)

Error

Per Cent

Two Term Equation

30

0.6715

0.009510

30.3

0.01983

0.3995

86.5

35

0.8251

—0.04203

30.2

0.1183

0.5076

109

45

0.08515

1.0004

55.2

Quadratic Term Only

30

0.6715

0.0000

28.6

0.01983

0.0000

142

35

0.8251

0.0000

28.5

0.1457

0.0000

144

45

1.0502

0.0000

0.08515

0.0000

141

Cubic Term Only

30

0.0000

0.009510

131

0.0000

0.3995

82.7

35

0.0000

—0.04203

136

0.0000

0.5378

114

45

0.0000

—2.6255

0.0000

1.0004

59.8

given for each fitted equation in Table 7. A slight modification was necessary in calculating the per cent deviation for nitroethane binary solutions. At all temperatures, the mean value of VE for each of these solutions was approximately zero. For tabulation purposes, and to eliminate anomalous values due to sign cancellation, the per cent devia¬ tion was calculated using the mean of the absolute values of VE.

The error columns of Table 7 veri¬ fy the conclusions drawn by Gunter, et. al. (1967) that the VE values of nitromethane are of the quadratic form obtained bv discard of all but the leading term of Equation (3) and the VE values of nitro¬ ethane are of the cubic form ob¬

tained by discard of all but the second term of Equation (3). For nitromethane, the two-term fit re¬ duces the error by a minimal amount compared to the use of only the quadratic term. In a like-manner, for nitroethane, the two-term fit is only slightly better than the use of only a cubic term. In every case, the relative smallness of one coeffi¬ cient in the two-term fit also indi¬ cates the validity of a single term fit. (The magnitude of the VE val¬ ues also changes the base used to calculate the error per cent which, in turn, explains the differences in magnitude of that quantity in Table 7.) The fact that CH3N02-CC14 solutions are less nearly ideal than C2H5N02-CC14 solutions is also sub-

80

Transactions Illinois Academy of Science

Table 8. — Summary of Literature Coefficients for Temperature Equations.

aQ

-ai x 103

a.2 x 106

Temperature Range

(g/ml)

(g/ml-°C)

(g/ml-(°C)2)

(°C)

Nitromethane

1.1574

1.356

20-256

1.1637

1.346

0-45 . 132

1.1639

1.340

0-2 524

1.1639

1.342

0-5023

1.1642

1.323

0-5027

1.1643

1.361

17. 3-60. 914

1.16448

1.351

20-306

1.1645

1.337

—1.15

0-1001

1.1646

1.377

16. 4-96. 326

1 . 1648

1.366

25-4516

1.1649

1.346

0-3025

1.1652

1.384

20-5029

1.1657

1.341

—4.94

0-8530

1.1657

1.377

20-3 07

1.1657

1.744

—5.49

—21. 5-101. 43

1.16576

1.383

25-602

1 . 1668

1.358

—0.55

20-1014

Nitroethane

1.06823

1.202

25-602

1.0707

1.210

18. 6-108. 526

1.0724

1.170

20. 2-87. 314

1.0743

1.187

20-307

1.0743

1.207

20-5029

1.0750

1.206

16. 6-79. 617

Carbon Tetrachloride

1.5239

—0.4607

—11.2

20-28331

1.6287

1.763

—2.09

8

1.6296

1.956

24. 2-54. 010

1.6306

1.949

11. 8-78. 028

1.6314

1.870

20. 1-59. 814

1.6321

1.877

—1.26

21

1.6325

1.920

0-40u

1.63255

1.9110

0.690

0-401

1.6326

1.920

20-2 5 12

1.6327

1.936

20-2 56

1.6329

1.927

—0.0469

1 5-7522

1.6329

1.927

0.563

10-8018

1.6331

1.945

0.392

15-7520

1.6334

1.961

11. 8-68. 09

1.6335

1.955

0.705

15-7519

1.6337

1.960

19. 35-58. 213

1.6347

2.006

25-4516

1.6468

2.759

15-4415

Musulin — Computerized Curve Fitting

81

1. Washburn (1928)

2. Boyd & Copeland (1942)

3. Jaeger & Kahn (1915)

4. Williams (1925)

5. Thompson, Coleman, & Helm (1954)

6. Dreisbach & Martin (1949)

7. Toops (1956)

8. Pugachevich, Nisel’son, Sokolova, &

Anurov (1963)

9. Renard & Guye (1907)

10. Morgan & Higgins (1908)

11. Cowley and Partington (1936)

12. Mumford and Phillips (1950)

13. Patterson & Thomson (1908)

14. Vogel (1948)

15. Souvek (1938)

16. Brown & Smith (1962)

stantiated by the tests for linearity given in Tables 3 and 4.

Nitromethane at 45° does not fit into the pattern of these calculations. The data which was obtained at 45° is such that the system of equations in the least squares procedure de¬ generates into a single equation. The result is that only the single constant, shown in Table 7, is de¬ rivable. In all other cases except one, the coefficient of the single term equation is the same as the corre¬ sponding term of the two-term equa¬ tion. This identity results from the facts that the terms occurring in the least squares procedure are sym¬ metric in x4 and x, and that the in¬ put data of the independent variables are symmetric in xx and x2. The sin¬ gle exception occurs when two data points are rejected, by the usual cri¬ terion (Worthing and Geffner, 1943), which destroys the symmetry of the input data.

Prigogine (1957) provides a theo¬ retical basis for Equation (3) through the use of the average po¬ tential model. The coefficients of Equation (3) can be estimated from the critical constants of the compon¬ ents of the binary solution. The critical constants of nitromethane (Weissberger, 1955) were used to make an estimate which could be

17. Ramsay & Shields (1893a)

18. Washington & Battino (1968)

19. Wood & Brusie (1943)

20. Wood & Gray (1952)

21. Fried and Schneier (1968)

22. Gibson & Loeffler (1941)

23. Walden (1909)

24. Walden (1906)

25. Timmermans & Hennaut-Roland

(1932)

26. Friend & Hargreaves (1943)

27. Walden & Birr (1933)

28. Ramsay & Aston (1894)

29. Geiseler & Kessler (1964)

30. Philip & Oakley (1924)

31. Ramsey & Shields (1893b)

32. Morgan & Stone (1913)

compared with the results in Table 7. Since the method depends upon which component of the solution is taken as the reference substance, the calculation was made in both frames of reference. The results are

VE = xlX2 [1.2898 + 0.001876(x1 —

x2)] CH3NO0 as reference

VE = Xlx2 [2.0960 + 0.003416 (x4-

x2)] CC14 as reference

Both estimates clearly show the same behavior as the results in Table 7, i.e. the quadratic term completely dominates the cubic term. Further, the order of magnitude of the quad¬ ratic coefficient is the same as those obtained in Table 7 (the estimation process is temperature independent) . Insufficient critical data for nitro- ethane prevented a similar estima¬ tion.

Acknowledgment

This research was supported by a grant from the Petroleum Research Fund (602- B) administered by the American Chemical Society. The authors gratefully acknowl¬ edge the use of the Data Processing and Computing Center of Southern Illinois University. Thanks are also accorded to the Cartographic Laboratories for the drafting work. In accordance with the obligation assumed with the use of the

82

Transactions Illinois Academy of Science

Choleski program, the authors wish to thank Eugene Fitzpatrick for permission to use, without fee, the subroutine for the calculation of the Student’s t-ratio in this “not for profit, scholarly research”.

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pp.

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Brown, I. and F. Smith. 1962. Volume Changes on Mixing. II. Systems Con¬ taining Acetone, Acetonitrile, and Nitro- methane. Australian J. Chem. 15, 9-12.

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Equilibria. VII. The Systems Nitro- methane + Benzene and Nitromethane + Carbon Tetrachloride at 45°C. Aus¬ tralian J. Chem. 8, 501-505. 5 tables.

Cowley, E. G. and J. R. Partington. 1936. Studies in Dielectric Polarisation. Part XX. The Dependence of Polarisa¬ tion and Apparent Moment of Nitriles LTpon Solvent and Temperature. J. Chem. Soc. 1184-1194. 6 tables, 3

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Daniels, F., J. W. Williams, P. Bender, R. A. Alberty, and C. D. Cornwell. 1962. Experimental Physical Chemistry. 6th Ed. McGraw-Hill Book Company, Inc., New York, xv + 625 pp. Dreisbach, R. R. and R. A. Martin. 1949. Physical Data on Some Organic Compounds. Ind. Eng. Chem. 41, 2875- 2878. 4 tables.

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Fried, V. and G. B. Schneier. 1968. Some Comments on Cohesion Energies of Liquids. J. Phys. Chem. 72, 4688-4690. 3 tables.

Friend, J. N. and W. D. Hargreaves. 1943. Viscosities and Rheochors of Ni¬ tric Acid, Nitroparaffins and their Iso¬ meric Nitrites. Phil. Mag. 34, 810-816.

2 tables, 2 figures.

Geiseler, G. und H. Kessler. 1964. Physikalische Eigenschaften Hemologer

Primarer und Stellungsisomer Gerad- kettiger Nitroalkane. “Nitro Com¬ pounds, Tadeusz Urbanski, Ed. The MacMillan Co., N.Y. 1964. Reprinted in Tetrahedron Supp. # 6, 20, 187-94. 5 figures, 2 tables.

Gibson, R. E. and O. H. Leoffler. 1941. Pressure- Volume-Temperature Relations in Solutions. V. The Energy-Volume Coefficients of Carbon Tetrachloride, Water, and Ethylene Glycol. J. Am. Chem. Soc. 63, 898-906. 8 figures,

1 table.

Gunter, C. R., J. F. Wettaw, J. D. Dren- nan, R. L. Motley, M. L. Coale, T. E. Hanson, and B. Musulin. 1967. Densities and Molar Volumes of Binary Solutions of Nitroparaffins in Carbon Tetrachloride. Jour. Chem. & Eng. Data. 12, 472-4. 3 tables.

IBM. 1962. IBM 1620 Fortran II Speci¬ fications. Pamphlet. International Business Machines Corporation. San Jose, Cal. 20 pp.

Jaeger, F. M. and J. Kahn. 1915. The Temperature Coefficients of the Free Molecular Surface Energy of Li¬ quids Between -80° and 1650° X. Meas¬ urements Relating to A Series of Ali¬ phatic Compounds. Proc. K. Akad. Wetensch. Amsterdam. 18, 269-285. Macfarlane, W. and R. Wright. 1933. Binary Liquid Systems and the Mixture Rule. Jour. Chem. Soc. 114-118. 2

tables.

Morgan, J. L. R. and E. Higgins. 1908. The Weight of a Falling Drop and the Laws of Tate. The Determination of the Molecular Weights and Critical Temperatures of Liquids by the Aid of Drop Weights, II. Jour. Am. Chem. Soc. 30, 1055-1068. 7 tables, 1 figure.

- ■. and E. C. Stone. 1913.

The Weight of a Falling Drop and the Laws of Tate. XII. The Drop Weights of Certain Organic Liquids and the Surface Tensions and Capillary Con¬ stants Calculated From Them. Jour. Am. Chem. Soc. 35. 1505-1523. Mumford, S. A. and J. W. C. Phillips. 1950. The Physical Properties of Some Aliphatic Compounds. J. Chem. Soc., 75-84.

Musulin, S. J. C. and B. Musulin. 1967. Ionization Energy as A Function of Nu¬ clear Charge. Trans. Ill. State Acad, of Sci. 60, 380-404. 4 tables.

Patterson, T. S. and D. Thomson. 1908. XXV. The Influence of Solvents on the Rotation of Optically Active Compounds. Part XI. Ethyl Tartrate in Aliphatic Halogen Derivatives. J. Chem. Soc. 93, 355-371. 4 figures.

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83

Philip, J. C. and H. B. Oakley. 1924. Conductivity and Ionisation of Solutions of Potassium Iodide in Nitromethane. J. Chem. Soc. 125, 1189-95. 6 tables. Prigogine, I. 1957. The Molecular Theory of Solutions. North-Holland Publ. Co., Amsterdam, xx + 446 pp. Pugachevich, P. P., L. A. Nisel’son, T. D. Sokolova, and N. S. Anurov. 1963. Density, Viscosity, and Surface Tension of Carbon and Stannic Tetra¬ chlorides. Zhur. Neorgan. Khim. 8, 791-6 cf. C. A. 60, 7483c (1964). Purcell, T. D. 1965. Least Squares Re¬ gression for the IBM 7040 by Ortho¬ normal Linear Functions Using the Choleski Method, Data Processing and Computing Center, Southern Illinois University, Carbondale, Illinois. Mimeo¬ graphed Materials. 45 pp.

Ramsay, W. and E. Aston. 1894. Die Molekulare Oberflachenenergie von Mis- chungen sich nicht associierender Flus- sigkeiten. Z. physik. Chem. 15, 89-97. 8 tables.

- . and J. Shields. 1893a.

LXXXI. The Molecular Complexity of Liquids. J. Chem. Soc. 63, 1089-1109. 2 tables, 1 figure.

- . und J. Shields. 1893b. Uber

die Molekulargewichte der Flussigkeiten. Z. physik. Chem. 12, 443-75. 14 tables.

Renard, T. et. P. -A. Guye. 1907. Measures de Tensions Superficielles a L’air Libre. Jour. chim. phys. 5, 8-112. 1 table.

Rowlinson, J. S. 1959. Liquids and Liquid Mixtures. Butterworths Scien¬ tific Publ., London, ix + 360 pp. Scatchard, G. 1949. Equilibrium in Non-Electrolyte Mixtures. Chem. Rev. 44, 7-35. 16 figures.

Soucek, B. 1938. Evidence of a Com¬ plex Between Nitrobenzene and Carbon Tetrachloride. Coll. Czech. Chem. Comm. 10, 459-65. 3 tables.

Thompson, C. J., H. J. Coleman, and R. V. Helm. 1954. The Purification and Some Physical Properties of Nitrome¬ thane. Jour. Am. Chem. Soc. 76, 3445- 6.

Timmermans, M. H. et Mine. Hennaut- Roland. 1932. Travaux du Bureau International d’Etalons Physico-Chem- iques. V. Etude des Constantes Phys¬ iques de Vingt Composes Organiques, J. chim. phys. 29, 529-568. UsolTveva, V. A. 1960. Classification of Density Diagrams for Liquid Binary Systems. Sb. Nauchn. Tr. Ivanovsk. Med. Inst. 23, 697-700. From: Chem. Abstracts 57, 1 3 2 2 3g (1962).

Vogel, A. I. 1948. Physical Properties and Chemical Constitution. Part XXIII.

Miscellaneous Compounds. Investiga¬ tion of the So-called Co-ordinate or Dative Link in Esters of Oxy-acids and in Nitro- Paraffins by Molecular Refractivity Determinations. Atomic Structural and Group Parachors and Refractivities. J. Chem. Soc. 1833-1855. 22 tables.

Walden, P. 1906. uber organische Lo- sungs-und Ionisierungsmittel. III. Teil Innere Reibung und deren Zusammen- hang mit dem Leitvermogen. Z. physik. Chem. 55, 207-249. 56 tables.

- . 1909. Ausdehnungsmodu-

lus, spezifische Kohasion, Oberflachen- spannung und Molekulargrosse der Lo- sungsmittel. Z. physik Chem. 65, 129“ 225. 67 tables.

- . and E. J. Birr, 1933. Leit-

fahigkeitsmessungen in Nitroverbindun- gen. 1. Leitfahigkeitsmessungen in Ni- tromethan. Z. physik. Chem. 163 A, 263-80. 1 figure, 35 tables.

Washburn, E. W., (Ed.). 1928. Inter¬

national Critical Tables McGraw-Hill Book Co., N.Y. xiv + 444 pp.

Washington, E. L. and R. Battino. 1968. Thermodynamics of Binary Solu¬ tions of Non-Electrolytes with 2,2,4-Tri- methylpentane. Ill Volumes of Mixing with Cyclohexane (10-80°) and Carbon Tetrachloride (10-80°). J. phys. Chem. 72, 4496-4502. 7 tables, 5 figures.

Weissberger, A. (Ed.) 1959. Technique

of Organic Chemistry. Vol. I-Part I. Physical Methods of Organic Chemistry. 3rd Ed. Chapter IV. Determination of Density by N. Bauer and S. Z. Lewin. Interscience Publishers, Inc., New York. 131-190 pp.

— . E. S. Proskauer, J. A. Rid¬ dick, and E. E. Toops, jr. 1955. Tech¬ nique of Organic Chemistry. Vol. VII. Organic Solvents. 2nd Ed. Interscience Publ., Inc., New York, vii + 522 pp.

Williams, J. W. 1925. A study of the Physical Properties of Nitromethane. Jour. Am. Chem. Soc. 47 , 2644-2652. 2 tables, 2 fig.

Wood, S. E. and J. P. Brusie. 1943. The Volume of Mixing and the Thermo¬ dynamic Functions of Benzene-Carbon Tetrachloride Mixture. J. Am. Chem. Soc. 65. 1891-5. 4 tables, 5 figures.

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The Volume of Mixing and the Thermo¬ dynamic Functions of Binary Mixtures. J. Am. Chem. Soc. 74. 3729-3733. 4

tables, 5 figures.

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Manuscript received September 29, 1970

ON VARIOUS METHODS USED IN THE CALCULATIONS OF INELASTIC ELECTRON-ATOM AND ELECTRON-MOLECULE COLLISIONS

YUH-KANG PAN

Department of Chemistry , Boston College, Chestnut Hill, Massachusetts 02161

Abstract. — Various approximate me¬ thods used in the calculation of inelastic electron-atom and electron-molecule col¬ lisions are classified into six different cate¬ gories, namely, Born approximation, Born- Oppenheimer approximation distorted wave Born approximation, distorted wave Born- Oppenheimer approximation, the method of integral equation, and variational method. The nature of these six different methods are described and their advantages and disadvantages are presented. The difficul¬ ties in the calculations of inelastic electron- molecule collisions are discussed. Some rec¬ ommendations are made for future progress in the calculation of inelastic electron- molecule collisions.

The theory of electron-atom and electron-molecule collision processes has been developed very rapidly in the last few years and has attained a very important practical meaning. This is because the best way to learn something about the forces acting between elementary particles is to observe their interaction with one another. Most of what we know about the microworld has been es¬ tablished by means of studying col¬ lision processes. Besides, the under¬ standing of a great many facets of chemical kinetics, spectroscopy, the physics of gaseous discharges, astro¬ physics, radiative transfer in flames and in the atmosphere, atomic phy¬ sics, physics of the upper atmosphere and solid state physics depends on our knowledge of the elementary in¬ teraction processes of electrons with atoms, molecules and other ele¬ mentary particles. The calculations

of inelastic electron-atom and elec¬ tron-molecule collisions are highly desirable. Because for many appli¬ cations it is necessary to have re¬ liable information on the rates of the various processes involving exci¬ tation of atoms and molecules by electrons. Many of the processes con¬ cerned are not readily investigated experimentally, so it is essential to rely to a considerable extent on theoretical prediction. Various ap¬ proximate methods have been used in the calculations of the inelastic scattering cross sections for electron- atom collisions. These methods can be roughly classified into six differ¬ ent categories, namely, Born ap¬ proximation, Born-Oppenheimer ap¬ proximation, distorted wave Born approximation (D.W.B.), distorted wave Born-Oppenheimer approxima¬ tion (D.W.O.B.), the method of in¬ tegral equations, and variational methods.

Born Approximation

The Born approximation assumes that the coupling of the incident electron with the atom is weak (all matrix elements of the interaction between the electron and atom, whether diagonal or non-diagonal are small). Therefore the calcula¬ tion of the scattering cross-section

can be carried out bv a first-order

«/

perturbation method. The effect of

[84]

Pan — Calculation of Inelastic Collisions

85

electron exchange between the inci¬ dent beam and the atom is ignored in this approximation. The cross section is then proportional to the square of the matrix element of the interaction between initial and finial states in which the free electron wave functions are plane waves and the overall wave function is simply an unsymmetrical product. The in¬ applicability of Bom approximation for electron energies near the thresh¬ old is well known (Bates, Funda- minsky and Massey, 1950). In low energy collisions the processes be¬ come quite complicated owing to various threshold effects which arise during the excitation and ionization of atoms and ions. Some transitions which are quite strong near thresh¬ old become completely forbidden in this approximation, because the effect of electron exchange is ignored in the approximation.

Born-Oppenheimer

Approximation

If we include the effect of electron exchange between the incident beam and the atom but still assumes that all matrix elements are small, then we obtain the so called Born-Oppen¬ heimer approximation to the scatter¬ ing amplitude or, in short, Born- Oppenheimer approximation (see, for example, B.L. Moiseiwitsch, Revs. Mod. Phys. 40, 238, (1968). However, we should distinguish this approximation from the Born-Op¬ penheimer approximation to the sep¬ aration of electronic and nuclear mo¬ tions). The overall wave functions used to calculate the probability am¬ plitude are properly summetrized combinations of plane wave and atomic wave functions of the form

'ko(ra)e,k<>n°,■ 'J'n ( ra ) e ik"r

where and are the wave func¬

tions of the initial and final atomic states respectively and ra represents the aggregate of the coordinates of the atomic electrons. The relative motion of the colliding electron is represented by undisturbed plane waves of wave length 27r/k0 before the collision and 27r/kn after the col¬ lision.

However, the Born-Oppenheimer approximation, apart from the in¬ clusion of the possibility of electron exchange, is still only a first-order perturbation formula. From an an¬ alysis of the observed data it appears that this approximation often over¬ estimates the importance of exchange to a very serious extent. This failure is particularly serious in the calcu¬ lations of the cross section near the threshold of excitation of one level from another if both belong to the same electron configuration and in other cases in which there is no change of azimuthal quantum num¬ ber of the atomic electron concerned in the process (e.g. when an s-s tran¬ sition is involved). Marriott (Mar¬ riott, 1958) has shown that the Born- Oppenheimer approximation cannot be relied on at all at low electron energies whether the coupling be¬ tween initial and final states is weak or not. Unfortunately, for many ap¬ plications it is the cross-section near the threshold which is required. Ochkur (Oclikur, 1963) thinks that the deficiencies of the calculations using the Born-Oppenheimer formu¬ la primarily result from an incor¬ rect extrapolation into the domain of low-energies. By using the Born- Oppenheimer formula as a basis, Ochkur obtains a new simple formu¬ la. The excitation functions have been calculated for the 23S and 23P level in helium by using this new formula. The results are in reason¬ ably good agreement with the experi¬ mental data. Rudge (Rudge, 1965)

86

Transactions Illinois Academy of Science

pointed out that Ochkur’s formula is not rigorous in the sense of being obtainable from a variational prin¬ ciple and gave an alternate expres¬ sion obtainable from a variational principle. Nevertheless, Ochkur’s formula has been successfully used in several other inelastic atomic col¬ lision calculations and has been ap¬ plied to the excitation of the hydro¬ gen molecule by Khare (Khare, 1966a, 1966b, 1967). Khare ’s results are in fair accord with experimental results and other theoretical esti¬ mates.

Distorted Wave Born Approximation

The distorted wave Born approxi¬ mation assumes that only the non¬ diagonal matrix elements are small. The plane waves which represent the initial and final free electron wave functions are then replaced by waves distorted by the mean interaction with the atom in the initial and final states respectively. It was first pointed out by Mott (Mott, 1932) that the contribution to the cross section for an inelastic collision be¬ tween two interacting systems which arises from impacts in which their relative angular momentum is il(l + 1) }1/2h can never exceed (21 {l l)X2/4 where X is the wavelength of the initial relative motion. Bates et al. have shown that the calculations for 0(23P, 2JS), 0+ (24S, 22D) and 02+(23P, 2*D and 2*S) using D.W.B. approximation yields cross sections in excess of the possible maxima (Mott, 1932). So it is also not a good method for the calculation of inelastic cross sections for electron- atom collisions.

Distorted Wave Born-Oppenheimer Approximation

The distorted wave Born-Oppen¬ heimer approximation also assumes

that only the non-diagonal matrix elements are small. The plane waves which represent the initial and final free electron wave functions are then replaced by waves F0 (r), Fn(r) dis¬ torted by the mean interaction with the atom in the initial and final states respectively. F„ (r) is the wave function which represents a plane wave eik°n°' and an outgoing spherical wave scattered by atom in its ground state, not allowing for the possibility of any inelastic col¬ lisions but including exchange ef¬ fects as far ts they effect elastic scat¬ tering. Fn(r) is the corresponding wave function representing a plane eil;,"' anci an outgoing spherical wave scattered by the atom in the nth excited state. The effect of elec¬ tron exchange in producing distor¬ tion is allowed for in this approxi¬ mation. Erskine and Massey (Ers- kine and Massey, 1952) first applied this approximation to the excitation of the 2S level of hydrogen from the ground state. They find that for electron energies near the threshold the cross section for the excitation is given by

Q = 4,kj|/3|2/k| (1)

kc and kx are the wave numbers of the initial and final motion of the election relative to the atom. \j3\ arises from requiring the solution of a pair of integro-differential equa¬ tions have the asymptotic form

f ~ ft exp(ikxr) (2)

Since the maximum possible value (Mott, 1932) for Q is 7r/k2, we may call 47rk1|^|2k0 the probability of the particular inelastic collision con¬ cerned.

The integro-differential equation expressed in atomic units is of the form

87

Pan — Calculation of Inelastic Collisions

[gp— 2V00 (r) +kl]i, + 5°X.

(r,r')f<. (r')dr' — 2V„ 1(r)f1(r) — t ( r,r' ) f t ( r' ) dr

[i-2V11(r) + K\]fx + fXi (r,r')£1(r')dr' = 2V.1(p)£.(r) -y?K1„(r,r')f. (r')dr' (3)

and the solutions must be proper functions satisfying the asymptotic conditions (2) and

fQ ~ Sink r + «exp(ik0r) (4)

In these equations Y00 and V1X are the interactions between the elec¬ tron and the atom arranged over the initial and final states of the atom, Koo and K1X are interaction kernels which represent the contri¬ bution of electron exchange to the mean interaction in each case, Vc x is the non-diagonal matrix element of the interaction and K01» Kxo are the corresponding contributions from exchange effects. If V0 K10 and K1C are zero the probability of the transition vanishes. If equation (3) is solved exactly the resulting cross section should be accurate, provided that the wave numbers kG and kx are sufficiently small so that inci¬ dent electrons with angular momen¬ ta greater than zero can be ignored and provided that the influence of other excited states is also negligible. It is quite clear that almost all of the violations of the conservation law (Mott, 1932) occur for electron energies near the threshold in which case the main contribution comes from head on collisions. The neglect of excited states is equivalent to ig¬ noring dynamic polarization effects. At the election energies concerned these are probably not very impor¬ tant, although it is difficult to esti¬ mate their order of magnitude. In

any case the exact solution of equa¬ tion (3) will certainly give cross sections which obey the conservation laws. It is only when the equation is solved by approximate methods that violation of these laws can oc¬ cur. All previous methods solve equation (3) by successive approxi¬ mations on the asumption that VQ K0 1 and K1C are small. The dis¬ torted wave Born-Oppenheimer me¬ thod makes no further assumptions, but the Born-Oppenheimer approxi¬ mation assumes that V 00-Vn/ K00 and K1X are also negligible and the Born approximation neglects Kxo and K01 as well. It is to be ex¬ pected that the D.W.B.O. method will give accurate results if Yc K01 and K10 are small but this is not sufficient to justify the Born- Oppenheimer approximation. In fact, for excitation of atoms by elec¬ trons with energy near the thresh¬ old the distortion introduced by Vco and VX1 is very marked. Ers- kine and Massey found their results for the excitation of the 2s level of atomic hydrogen were considerably different from those obtained by use of the Born-Oppenheimer approxi¬ mation in which distortion is ne¬ glected. Whereas the latter approxi¬ mation gives cross-sections at ener¬ gies close to the threshold which are greater than the maximum pos¬ sible value, the D.W.B.O. method gives values always less than this. Nevertheless, it approaches within 30% of this value at low energies. The coupling between the motion in the initial and final states is quite strong in this case. The D.W.B.O. approximation, which depends on this coupling being weak, cannot be expected to yield very good results under these conditions. Further evi¬ dence in support of this was obtain¬ ed in a calculation, by a variational method, carried out by Massey and

88

Transactions Illinois Academy of Science

Moisewitsch (Massey and Moise¬ witsch, 1953), which did not require the coupling to be weak.

The next application of the D.W.B.O. method was to the excita¬ tion of the 23S and 2XS states of helium by Massey and Moiseiwitsch (Massey and Moiseiwitsch, 1954) . In the 23S calculation they found a resonance effect very close to the threshold, leading to a sharp peak in the cross section, and the absolute magnitude was much smaller than given by the Born-Oppenheimer ap¬ proximation. This was due to the fact that the distortion effectively annihilated the partial cross-section for excitation by incident electrons of zero angular momentum. Their results are in agreement with the experimental data, and represent an important success of the method. It is noteworthy that in this case the cross sections are all well below the maximum possible values showing the coupling to be weak. In the 2XS state, experimental results show that a sharp peak in the excitation func¬ tion near the threshold also occurs. This does not appear according to their calculations probably because the close coupling between the 23S and 2XS state is ignored. Evidence for this has been afforded by accur¬ ate numerical calculations by Mar¬ riott on the cross-section for the de¬ activation of 2XS helium atoms by slow electrons which produce transi¬ tions to 23S states. The first appli¬ cation of the D.W.B.O. method to an s-p transition was made by Kha- shaba and Massey (Khashaba and Massey, 1958) for the excitation of the 2p level of hydrogen. They cal¬ culated the total cross-section and the polarization of the radiation ex¬ cited by the impact. Their results did not differ very much from those given by the Born-Oppenheimer ap¬ proximation either for the total

cross-section or for the polarization, but this was partly coincidental. Thus the distortion virtually an¬ nihilates the partial cross-section qc (for scattered electrons with zero angular momentum) but largely compensates this by increasing qx (for scattered electrons with angu¬ lar momentum \/2h). The usual tests of coupling indicate that it is not very strong (the distorted wave partial cross-section never ex¬ ceeds one-quarter of the maximum possible value 37r/k2). Massey and Moiseiwitsch carried out a calcula¬ tion of the cross-section for excita¬ tion of the 23P state of helium by D.W.B.O. method on similar lines to those of Khashaba and Massey (Khashaba and Massey, 1958) for the 2p of hydrogen. The triplet state calculation is of great interest because excitation can only come through exchange which limits the significant contributions to the first two partial cross-sections for which the effects of distortion are much more marked. In other words, there is less dilution of these effects from the higher order partial cross-sec¬ tions which are little affected by distortion. Their results have been compared with the corresponding re¬ sults given by the Born-Oppenheim¬ er approximation. Although there are considerable differences at elec¬ tron energies below lOOeV, these are much less marked than for the exci¬ tation of the 23S state (Massey and Moiseiwitsch, 1954). The D.W.B.O. method gives cross-sections in closer agreement with observation but sub¬ stantial discrepancies still remain. The coupling between the 1XS and 23P states is nowhere as strong as judged by the ratio of their results for the partial cross-sections to the maximum possible value. Thus their calculated values for either qQ or qa is never as great as 0.25 of the

Pan — Calculation of Inelastic Collisions

89

maximum and is usually much less. Effects due to coupling' between 2P and 28 states are likely to be smaller

t

than for atomic hydrogen owing to the larger energy differences. The cross-section for excitation of the 2p state of hydrogen by electron im- pact, calculated by Kliashaba and Massey (Kliashaba and Massey, 1958) using the exactly similar D.W.B.O. method, is in consider¬ ably closer agreement with the ob- servations of Fite and Brackmann (Fite and Brackmann, 1958). On the other hand, in the hydrogen case, the atomic waye functions are ac¬ curately known and no account has to be taken of the coupling between 23P and 2XP states. It is difficult to judge how important these two fac¬ tors are. It is also difficult to esti¬ mate the accuracy of Massey and Moiseiwitsch ’s results with any cer¬ tainty because there are inconsisten- cies in the observed data. In any case, one would have expected the D.W.B.O. method to give very good results if the coupling between the motion in the initial and final states is weak.

The Method of Integral Equations

The method of integral equations had been developed by Drukarev (Drukarev, 1953). Using this meth¬ od, the excitation of the sodium atom by slow electrons has been calcu¬ lated (Veldre, 1956). In this cal¬ culation only the s-wave of the inci¬ dent beam was taken into account. For a certain value of incident elec¬ tron energy the exchange and strong coupling have been included. The elastic scattering of slow electrons by lithium atoms with the inclusion of exchange was calculated by the-

Drukarev approximation (Veldre, Gailitis, Damburg and Stepinsh, 1956). In the incident beam only the s-wave was taken into account. The question of the choice of radial wave functions of atomic electrons was investigated (Veldre, 1959) and it was shown that the behavior of the radial wave function near the zero does not have an important in¬ fluence of the magnitude of the ef¬ fective cross-section. In the inelas¬ tic cases the main advantage of the method of integral equations is that it can obtain analytical expressions for the desired atomic wave func¬ tions of different states for all r, and not merely their asymptotic forms. This feature can be used to obtain appropriate classes of varia¬ tion functions associated with the calculation of electron scattering by variational methods. Matora (Ma- tora, 1960) applied the Drukarev method to calculate the elastic s- scattering of electrons and the ex¬ citation functions of the 23S and 2XS states of helium by electrons with energies from 0 to 40 eV. How¬ ever, Massey and Moiseiwitsch ’s (Massey and Moiseiwitsch, 1954) re¬ sult for 23S calculated by D.W.B.O. methods corresponds more closely to the experimental data than the re¬ sult obtained by Matora using the method of integral equations. Dam¬ burg et al., pointed out that the method of integral equations has slow convergence. The effective cross-section for the elastic scatter¬ ing of electrons by hydrogen atoms, calculated by the second approxima¬ tion of the method of integral equa¬ tions, agrees with the effective cross- section calculated by the second Born approximation. However, cal¬ culations by the second Born ap¬ proximation are less complicated than the method of integral equa¬ tions.

90

Transactions Illinois Academy of Science

Variational Method

As mentioned before, it seems D.W.B.O. method is a reasonably good approximation in inelastic scattering of electrons by atoms and molecules. Owing to the labor in¬ volved in calculating the distorted waves F0 and Fu the method can¬ not be applied widely. However, variational methods greatly facili¬ tate D.W.B.O. calculations, for both the functions F0 and Fn may be obtained in a convenient analytical approximation for both the antisym¬ metric and symmetric cases.

The application of variational methods to bound state problem has proved to be very fruitful, especial¬ ly for obtaining approximations to the lowest eigenvalue of the energy. Although in principle still applica¬ ble to the approximate determination of the energies of excited states, the method becomes much less conven¬ ient because of the volume of analy¬ tical and computational work re¬ quired. Variational methods for dealing with atomic collision prob¬ lems were first proposed by Hulthen (Hulthen, 1944). Hulthen proposed a method, based on his variational principle, for the approximate cal¬ culation of the radial function and its phase, and verified his method on the simplest examples. In 1947, Schwinger (Schwinger, 1947) devel¬ oped a variational method, different from Hulthen ’s method, which was based on an integral equation for the wave function. In 1948, Kohn (Ivohn, 1948) geenralized Hulthen ’s formulation, extending it to the gen¬ eral case of scattering. Following on his work, a series of papers ap¬ peared (see for example, Newton, 1966) in which new variational me¬ thods were proposed. However, all these methods do not differ essen¬ tially from the two basic methods :

the Hulthen-Kohn method based on Schrodinger ’s differential equation and Schwinger’s method based on an integral equation. These varia¬ tional methods are similar to those used for bound state problems but they differ in certain important re¬ spects. In both cases the aim is to obtain expressions involving wave functions describing the state of sys¬ tems which remain correct to the first order when a variation is im¬ posed on one or more of these func¬ tions. The bound state energies are found to be true minima with re¬ spect to the variational calculations under the imposed conditions while this is not true for problems of un¬ closed states which described col¬ lision processes. This has the con¬ sequence that greater flexibility in the choice of trial functions may even lead to less satisfactory re¬ sults, a situation which can never arise in bound state problems. Much greater care has to be taken there¬ fore in applying variational meth¬ ods under these circumstances and it is usually difficult to estimate the accuracy of the results. The applica¬ tion of variational methods of scat¬ tering problems has also been lim¬ ited by the complexity of the inte¬ gration required in the use of the Schwinger formulation even for the simplest trial functions, and the dif¬ ficulty of finding adequate trial functions in the relatively simple Kohn formulation. In spite of the difficulty of applying variational methods to collision problems, ex¬ tensive development of the theory of elastic (Massey and Moiseiwitsch, 1950; Moiseiwitsch, 1953) and in¬ elastic scattering of electrons by atoms using the variational methods of Hulthen and of Kohn has been carried out. The distorted wave functions used in Erskine and Mas¬ sey’s (Erskine and Massey, 1952),

Pan — Calculation of Inelastic Collisions

91

Massey and Moiseiwitsch ’s (Massey and Moiseiwitsch, 1953), Khashaba and Massey’s (Khashaba and Mas¬ sey, 1958), and Massey and Moisei¬ witsch ’s calculations (Massey and Moiseiwitsch, 1960) are determined by Hulthen ’s and Kohn ’s varia¬ tional methods. The results obtained have been encouraging. The gener¬ alization of Hulthen ’s variational method to the inelastic scattering of electrons by atoms was first derived by Moiseiwitsch (Moiseiwitsch, 1951) by using the integral L = ^#(H-E)^dr where H is the ham- iltonian of the system. The complex

parameters a, d, cx . . cn in the

trial function are determined through the conditions

Lt = 0 (5)

0Lt kx 0Lt

- [- 2i — d* — = 0 (6)

0a k 0a

0Lt

- — 0, (i— 1, . n) (7)

0Ci

where Lt = ^ 'I't* ( H-E) 'Jqdr (8)

and a and d are complex. In con¬ trast to Hulthen ’s variational meth¬ od applied to the elastic (Moisei¬ witsch, 1953) scattering of electrons, the condition L=0 does not imply that a=0. A correction to the par¬ ameter may be obtained by consid¬ ering the integral

L' = j*(H-E)*dr (9)

For it can be shown that

81/ = 47rk8a (10)

therefore the corrected value of the parameter a is given by

A = a — L'/47rk (11)

This variational method can be ex¬ tended to include any order partial wave, and excitation to any state of the atom. Massey and Moiseiwitsch (Massey and Moiseiwitsch, 1953) first applied this method to the cal¬ culation of the ls-2s electron exci¬ tation cross-section of hydrogen. Their results are qualitatively more in accord with those of the accurate numerical method of Marriott (Mar¬ riott, 1958) than either of the other methods. The poor quantitative agreement is probably a consequence of the use of an over-simplified trial function in the variational calcula¬ tion. It is apparent that this varia¬ tional method does not require the coupling between the motion in the initial and final states to be weak and the results of this method are better than those of other approxi¬ mations. However, the great diffi¬ culty in applying this variational procedure is the complexity of the calculations involved even when us¬ ing the simplest trial functions. For this reason, in Massey and Moisei¬ witsch ’s calculation of the ls-2s electron excitation cross-section of hydrogen, detailed numerical work was confined to trial functions of the form

f0 = Sin kr + (A -(- be‘r)

(1 — e*r) cos kr (12)

fx = (1 — er)d exp(ikxr) (13)

b being the additional variable parameter. These wave functions suffer from the defect that they do not allow for mixing between the incident and scattered waves. Even for such over-simplified trial func¬ tions the analysis involved, is very extensive and the determination of “a” from the equations (5) to (7) was quite involved.

92

Transactions Illinois Academy of Science

Electron-Molecule Collision Problems

Although the scattering of slow electrons by atoms has been exten¬ sively studied theoretically, com¬ paratively little theoretical work has been done on the elastic and inelas¬ tic scattering of slow electrons by diatomic or polyatomic molecules. This is undoubtedly due to the math¬ ematical complexity of the problem. We cannot apply Born’s approxima¬ tion, because it is least reliable in the low energy domain. Thus we have to solve the Schrodinger equa¬ tion for the incident electron direct¬ ly. This problem is fairly simple in the case of atoms on account of the sperical symmetry of the poten¬ tial field, but in the case of mole¬ cules, the molecular potential (force) is not spherically symmetric, so the Schrodinger equation is not separ¬ able and the analysis becomes very complicated. Under certain condi¬ tions, however, it is possible to treat the individual atoms in the molecule as independent scattering centers so that the amplitude for scattering by the individual atoms in the mole¬ cule can be obtained by adding the amplitudes for scattering by the in¬ dividual atoms with proper allow¬ ance for phase differences. The re¬ sulting cross sections must then be averaged over all molecular orienta¬ tions to give observable data. This appoximation will only be valid if multiple scattering of an electron within the molecule is negligible and if the distortion of the atomic fields by the valence forces is also unim¬ portant. Both these conditions are likely to be Avell satisfied for impacts with fast electrons for which Born’s first approximation gives an ade¬ quate representation of the atomic scattering (Massey, 1956). It is not so obvious that when Born’s first

approximation is inadequate the as¬ sumption of independent scattering will always be satisfactory, but there is evidence that it does have a range of validity extending to scattering of electrons with veloci¬ ties much too slow for Born’s first approximation to be applicable. On the other hand, for very slow elec¬ trons, with wave-lengths comparable with the atomic separations in the molecule, there is no doubt that the independent scattering approxima¬ tion is no longer valid and the cal¬ culation of the scattering presents much greater difficulties.

According to our discussions in the above sections, it seems more flexible to use the variational meth¬ od for inelastic scattering of the elec¬ trons by diatomic or polyatomic molecules. The behavior of phases at small energies, the relation be¬ tween the discrete and the continu¬ ous spectrum are directly or indi¬ rectly connected with variational principles. Besides, it is well known that the basic equations of stationary perturbation theory for bound state can be derived from a variational principle. Similar results can also be obtained in collision theory for the scattering amplitudes and the phases if one starts from the sta¬ tionary property of appropriate functionals. As regards numerical calculations, it seems as if only vari¬ ational methods allow one to take into account effectively and rigor¬ ously such phenomena as the polari¬ zation of an atom by an incident electron and obtain results of the same degree of accuracy as is at¬ tained in the evaluation of atomic and molecular energy levels. How- ever, in order to generalize Moisei- witsch’s variational method for in¬ elastic scattering of electrons by molecules, first we have to solve the difficulties of the variational meth-

Pan — Calculation of Inelastic Collisions

93

od (such as the complexity of the calculations involved and the ques¬ tion of choosing trial functions). As to the trial functions, the Hulthen’s, Kohn’s and Moiseiwitsch ’s criteria for selection of the “best” values of the parameters are reasonable, but of course not essential or neces¬ sarily the best to choose. Hulthen’s and Kohn’s criteria merely make the trial function simulate one prop¬ erty of the exact wave-function. We shall examine various alternative procedures and look for the possi¬ bilities for finding the best criteria. The special feature of the method of integral equations as mentioned before shall be studied and the ap¬ plicability to obtaining appropriate classes of variation function asso¬ ciated with calculation of electron inelastic scattering by the variation¬ al method shall be examined. For diatomic molecule calculations, the difficulty due to the distortion of a two-center field also should be over¬ come. For inelastic scattering of electrons by the hydrogen molecule, Huzinaga’s (Huzinaga, 1957) and Hoyland’s (Hoyland, 1966) one-cen¬ ter wave functions shall be used for both the initial and final states of the molecule. This will allow all the integrals to be evaluated exactly and with little labor. Calculations on this line are now in progress in our laboratory.

Acknowledgments

The author is indebted to Herschel Rabitz of Harvard University for reading the manuscript.

Literature Cited

Bates, D. R., A. Fundamisky, J. W.

Leech and H. S. W. Massey, 1950.

Excitation and Ionization of Atoms by

Electron Impact. Phil. Trans. 243 A:

93-143.

Damburg, R. and V. Kravchenko, 1960. The Scattering of Electrons By Alkali Metal Atom. V. Latv. PSR Zinat. Akad. Vest. 1:73-81.

Drukarev, G. F. 1953. Excitation of the Sodium Atom By Slow Electrons. Zhur. Eksptl. Theoret. Fiz. 25: 129-

133.

Erskine, G. A. and H. S. W. Massey, 152. The Application of Variational Method to Atomic Scattering Problems II. Proc. Roy. Soc. 21 2A: 521-530.

Fite, W. N. and R. T. Brackmann. 1958. Collision of Electrons With Hydrogen Atoms II. Physi. Rev. 112: 1151-1156.

Hoyland, J. S. 1966. Single-Center Cal¬ culations on the Lowest-Lying va and tu Excited States of H2. J. Chem. Phys. 45: 3928-3933.

Hulthen, L. 1944. Neutron-Proton Scattering in the Region 0-5 Mev. Fysio- gr. Sallsk. Lund. Forhandl. 14: 2-8.

Huzinaga, S. 1957. One-Center Expan¬ sion of Molecular Wave Function, II. Progr. Theoret. Phys. (Kyoto) 17: 162- 168.

Kare, S. P. 1966a. Excitation of Hydro¬ gen Molecules by Electron Impact. Phys. Rev. 149: 33-37.

- , 1966b. Excitation of Hydro¬ gen Molecules by Electron Impact II. Phys. Rev. 152: 74-75.

- , 1967. Excitation of Hydro¬ gen Molecule by Electron Impact III. Phys. Rev. 157: 107-112.

Khasaba, S. and H. S. W. Massey. 1958. The Excitation of the 2p State of Hydro¬ gen by Slow Electrons — Distorted Wave Treatment. Proc. Phys. Soc. 71 A: 574-584.

Kohn, W. 1948. Variational Methods in Nuclear Collision Problems. Phys. Rev. 74: 1763-1772.

- , 1949. Variation Scattering

Theory in Momentum Space I. Phys. Rev. 84: 495-501.

Marriott, R. 1958. Calculation of the ls-2s Electron Excitation Cross Section of Hydrogen. Proc. Phys. Soc. 72 A: 121-129.

Massey, H. S. W. 1956. Theory of Atomic Collisions. Handbuch der Physik, 36: 232-408.

Massey, H. S. W. and B. L. Moisei¬ witsch. 1950. The Application of Variational Methods to Atomic Scatter¬ ing Problems I.

_ 3 1953. Calculation o f ls-2s

Electron Excitation Cross Section of Hydrogen By a Variational Method. Proc. Phys. Soc. 66A: 406-408.

94

Transactions Illinois Academy of Science

- , 1954. The Application of

Variational Methods to Atomic Scatter¬ ing Problems IV. Proc. Roy. Soc. 227A: 38-51.

- , 1960. The Excitation of the

23P State of Helium By Electron Impact. Proc. Roy. Soc. 258A: 147-158.

Matora, I. M. 1960. The Scattering of Slow Electrons By Helium Atoms, With Excitation of the 23S and 2’S Levels. Opt. i. Spectr. 9: 707-713.

Moiseiwitsch, B. L. 1951. A Varia¬ tional Method for Inelastic Collision Problems. Phys. Rev. 82: 753-753.

- , 1953. The Application of

Variational Methods to Atomic Scatter¬ ing Problems III. Proc. Roy. Soc. 219A: 102-109.

Newton, R. G. 1966. Scattering Theory of Waves and Particles. McGraw-Hill Book Company, New York.

Mott, N. F. 1932. The Theory of the Effect of Resonance Levels on Artificial

Disintegration. Proc. Roy. Soc. 133A: 228-240.

Ochkur, V. I. 1963. The Born-Oppen- heimer Method in the Theory of Atomic Collisions. Zhur. Eksptl. Theoret. Fiz. 45: 734-741.

Rudge, M. R. H. 1965. The Calcula¬ tion of Exchange Scattering Amplitudes. Proc. Phys. Soc. 85: 607-608.

Schwinger, J. 1947. A Variational Principle of Scattering Problems. Phys. Rev. 72: 742-742.

Veldre, V. 1959. Excitation of Atoms by Electrons. Latv. PSR. Zinat. Akad. Vest. 5: 105-109.

- , M. Grailitis, R. Damburg

and Stepinsh. 1956. Elastic Scat¬ tering of Slow Electrons by Lithium Atoms. Latv. PSR. Zinat. Akad. Vest. 5: 73-81.

Manuscript received April 6, 1970

NOTES

WHAT EFFECT DOES PROLONGED FLOODING HAVE

ON ANT COLONIES?

C. A. DENNIS, D. HENRY, D. WALCH, G. JONES, AND L. C. SHELL

Department of Biology, Millikin University, Decatur, Illinois

Abstract. — This study shows that prolonged flooding eliminated all species of ants, in the area worked, with the excep¬ tion of Camponotus pennsylvanicus and Camponotus ( Myrmentoma ) nearticus. It is suggested that in some manner these spe¬ cies are able to block off the water from their nests.

This question came to the mind of the senior author after observing for several years the prolonged and frequent flooding of a tract of woodland along the Sangamon River. The area is flooded to a depth of several feet for from several days to several weeks during the spring and sum¬ mer, and it seems reasonable to think that all the ants in this area except those living in trees would be drowned. If any colo¬ nies were found below flood level during summer or fall they would be the result of fertile queens moving in and establishing colonies after the water receded.

Collecting in August of 1967 showed six species. All the colonies with the excep¬ tion of Camponotus pennsylvanicus and Camponotus (Myrmentoma) nearcticus were small, but the colonies of the two spe¬ cies of Camponotus were large and vigorous. In early May of 1968, after severe flooding the area was again searched. Only colo¬ nies of the species Camponotus were found. These were located in very large and soggy

logs within a few feet of the rivers edge. This discovery led us to believe that in some manner these two species had sur¬ vived after being submerged for a long period of time.

In the fall of 1968 the area was again covered, with practically the same results as August of 1967. Then in March of 1969 after the area had been flooded the results were the same as 1968.

There seems no doubt that these species are able to withstand being submerged for a long time. The area of these logs men¬ tioned is some yards away from any stand¬ ing timber and as rapid as the current is at this place it seems impossible for a large colony to be able to swim to safety of standing trees. Another factor was noticed which led us to believe the colonies did not move out. The logs were sopping wet everywhere with the exception of the region of the colonies.

We propose to continue the investigation to find, if possible, the method used by these two species to avoid drowning when flooded.

Acknowledgement

We are indebted to Dr. William S. Creighton for checking the identification of the species.

Manuscript received July 7, 1970

[95]

TRIBUTE TO DR. JAMES W. NECKERS

RICHARD T. ARNOLD

Department of Chemistry, Southern Illinois University, Carhondale 62901

It is rare enough for a young scientist to spend four decades in continuous service as a teacher at one institution before re¬ tirement, and even of this small group, few can look back on such a remarkable degree of change — much of which they promoted — as can Dr. James W. Neckers. On Saturday, October 10, 1970, Southern Illinois University, in grateful recognition of his superb contributions, dedicated its new six-million dollar Physical Science Building and officially named it the James W. Neckers Building.

Dr. Neckers, a native of New York, was graduated from Hope College; he com¬ pleted his Ph.D. at the University of Illinois in 1927, at the age of 25, and joined the faculty at S.I.U. Two years later, he took over the chairmanship of the Depart¬ ment of Chemistry. Dr. Neckers played a leading role in the design of the new Parkinson Laboratory. But most important to those of us who were chemistry majors was the fact that we were exposed to enthusiastic, well-trained and dedicated teachers who devoted an enormous amount of time encouraging and counseling each of us.

At that time, the Chemistry faculty con¬ sisted of only four members (known affec¬ tionately as the “Four Horsemen”) ; name¬ ly, Drs. Neckers (Analytical-Inorganic), T. W. Abbott (Organic), R. A. Scott (Bio¬ chemistry) and K. A. Van Lente (Phyical). Although each man taught the advanced courses in his respective specialty, all co¬ operated in the teaching of Freshman Chemistry. It was not until 1945 that a fifth man was added to the staff, and shortly afterwards, an astounding rate of growth ensued.

When Dr. Neckers came to S.I.U., the institution conferred only the B.Ed. degree and consisted of a total student body numbering less than 2,000. At the time of his retirement (1967), S.I.U. had be¬ come a full-fledged university with over 20,000 students, a large graduate school with doctoral programs in many disciplines and a chemistry faculty of 24 members. Under his guidance, an ACS accredited program for Chemistry majors was ap¬ proved, and, subsequently, M.S. and Ph.D. programs were started in 1957 and 1963, respectively.

Dr. Neckers served as a President of the Illinois Academy of Science, and was long active as a member of the Illinois Teachers Association, National Teachers Association, the American Chemical Society and other professional organizations.

His role, however, was not limited to things chemical and scientific. As a true scholar, his concern for the welfare of the entire University was great. He was active in A.A.U.P. and a leader in faculty participation in University affairs which led to the formation of the Faculty Council.

In 1966, former students elected him as recipient of the Alumni Association’s Great Teacher Award. Many of these alumni do not know that he was active in the planning of the new building which now bears his name. They remember best his quick wit, his dry humor, his absolute dedication to high standards of scholarship and personal behavior and his untiring effort to impress these upon every student who was privileged to be associated with him. This is why hundreds of former students admire and respect him both as teacher and friend.

[96]

A CONVENIENT METHOD FOR THE PREPARATION OF

AROMATIC a-DIKETONES

JERRY HIGGINS AND JOE F. JONES

Illinois State University, Normal, Illinois

Abstract. — A convenient method has been developed for the preparation of aro¬ matic a-diketones from a-halo aromatic ketones and pyridine-N-oxide. Desyl chlor¬ ide (1) and phenylglyoxalybenzil (4) were prepared in 50% and 70% yields, re¬ spectively.

A new synthesis of glyoxals and a-dike¬ tones has recently been described (Korn- blum, et al. 1966). This synthesis involves the reaction of a-halo ketones with nitrate to produce the organo nitrate esters fol¬ lowed by the elimination of nitrite with a weak base such as sodium acetate. Ben- zil was obtained in 95% yield from desyl bromide and yields of various glyoxals ranged from 82-90%. A similar method which has been used to form aromatic aldehydes from the reaction of benzyl hal¬ ides and pyridine-N-oxide has been re¬ ported in the literature (Feely et al. 1957). In this particular reaction pyridine-N-oxide is used as the nucleophile for producing benzyloxypyridinium bromide from benzyl bromide. This salt is then treated with dilute sodium hydroxide to produce benzal- dehyde by the abstraction of a proton from the benzyl carbon and the elimination of pyridine. Yields of benzaldehyde and aro¬ matic dialdehydes ranged from 80-95%. The chemical requirements for reactions of this type are that a nucleophile capable of displacing reactive halides is used and that this nucleophile possesses a good leaving group with the elimination of a proton in the formation of the carbonyl group.

The method we would like to report in this communication is a combination of the procedures used by Feely and Kornblum. Our modified method uses aromatic a-halo ketones as in Kornblum’s method and pyri- dine-N-oxide as in Feely’s method.

Experimental

Preparation of Benzil(2). — Equal molar quantities of desyl chloride and pyridine-N- oxide were refluxed for three hours in acetonitrile. An equal molar quantity of

+ <oVo %H.3.C°2N

Cl w CH3CN

{o)"C-c-(o) + (Uji

<O>?-0H^O>CH?Xo) +

Br Br

1

ch3co2 Na CH3CN

_ 00^00„ ^ <§>c!-c^>c-c^o) 4- 2 (op

4

Figure 1. Scheme for Synthesis of a- diketones.

sodium acetate was then added, and the reaction mixture was refluxed for an addi¬ tional three hours. After removal of two- thirds of the acetonitrile, the reaction mix¬ ture was poured into water and the product collected by filtration. Recrystallization from methanol gave 50% yield of benzil melting at 91-94°C (reported m.p. 95°).

Preparation of a a'-dibenzoyl- a, a'- dibromo-/?-xylene (3). — to 150 ml of glacial acetic acid was added 11.6 g (0.037 moles) of a, a'-dibenzoyl-/?-xylene (Wrasidlo and Augl, 1969). After dissolv¬ ing the compound, 12.5 g (0.078 moles) of bromine was added dropwise to the warm reaction solution. The product was collected by filtration and washed three times with 100 ml portions of water. The yield of crude product was 15 g (86%). After recrystallization from carbon tetra¬ chloride, 10.9 g (62%) of pure product, m.p. 179-180°C, was obtained. Calcd. for 02HisBr20: C, 55.94%; H, 3.42%; Br, 33.85%. Found: C, 55.83%; H, 3.21%; Br, 34.00%.

Preparation of phenylglyoxalylbenzil(4) .

[97]

98

Notes

— To 200 ml acetonitrile were added 25.0 g (0.053 mole) of a, a’-dibenzoyl- a, a’-dibromo-^-xylene and 12.0 g (0.126 mole) pyridine-N-oxide. The re¬ action solution was refluxed for three hours and then 12.0 g of anhydrous sodium ace¬ tate was added to the hot solution. After refluxing for an additional three hours, about two-thirds of the acetonitrile was removed and the remaining residue added to 300 ml of water. The aqueous mixture was stirred for 30 minutes. The product was filtered oflf and then dried in vacuo. Recrystallization from methanol gave 12.3 g (0.036 mole) (70%) of product melting at 124-126°C. Reported m. p. 126°C (Oli- arinso, et al. 1963).

Results and Discussion

Benzil and phenylglyoxalylbenzil can be prepared easily and conveniently by the reaction of pyridine-N-oxide with aromatic a-halo ketones in acetontirile solvent (Scheme 1). Other solvents such as alco¬ hols, cyclic ethers, diols, N,N-dimethylace- tamide were also used as solvents in the present procedure, and the yields were

similar to those in acetonitrile solvent. Since this appears to be a general reaction, we are presently expanding this procedure to the preparation of glyoxals and other a-diketones.

Literature Cited

Feely, W., W. L. Lehn, and V. Boekel- heide. 1957. Alkaline Decomposition of Quarternary Salts of Amine Oxides, /. Org. Chem. 22 , 1135,

Kornblum, Nathan, and H. W. Frazier. 1966. A New and Convenient Synthesis of Glyoxals, Glyoxalate Esters, and oc- Diketones. J. Am. Chem. Soc. 88, 865- 866.

Ogliarinso, M. A., L. A. Shadoff, and E. Becker. 1963. Bistetracyclones and Bishexaphenylbenzenes. J. Org. Chem. 28, 2725-2728.

Wrasidlo, W., and J. M. Augl. 1969. Phenylated Polyquinoxalines from 1,4- Bis (phenylglyoxaloyl) benzene. J. Poly¬ mer Sci. B7, 281-286.

Manuscript received October 1, 1970

RE: HAWKINS AND KLIMSTRA, “DEER TRAPPING CORRELATED WITH WEATHER FACTORS”

(TRANS., 63:198-201)

H. W. NORTON

Department of Animal Science, University of Illinois, Urbana, Illinois 61801

The use of factor analysis, probably al¬ ways dubious, is exemplified in its uncer¬ tainties by application separately to data for two different years. After the inde¬ pendent variables were “loaded on 12 fac- ors”, selection of “the independent variable under each factor that contributed most” led to two sets of 12 variables having only a single variable (number 17) in common. This is significantly (P = 0.017) less agree¬ ment than should occur by chance (though there is at least one misprint affecting variables “deleted from 1964-65 trap period” without explanation). Can the authors still believe that the factor analy¬ sis serves “to eliminate redundant factors”? If so, must they not conclude that all factors were probably “redundant”? Fur¬ thermore, this common variable failed to exhibit a significant correlation with trap success in the first year (no similar state¬ ment was made about data for the second year) .

Of the four independent variables which exhibited “significant t-test values” in the first year, it is said that only two “had significant r values”. If this means partial correlation, the test of correlation coeffi¬ cients should yield results identical to those from the t test; if total correlation is meant,

the analysis is regressing from a more detailed and specific result to a relatively crude result.

The authors remark that, although 6 corral and 3 box traps were available, they were not always all in use and that data for nights with only one or two traps set were ignored. However wise that may be, surely the analysis should have taken account of the individuality of the traps, not to mention the possible difference be¬ tween the two types. It may be that per¬ manent differences between traps contrib¬ ute so much to the variance relegated to “error” in the reported analysis as to ob¬ scure completely real effects of some of the weather variables studied.

The data should be reanalyzed taking explicit account of trap individuality, and involving data for the two years so as to test whether years are in reasonable agree¬ ment. Those of your readers who are interested in factor analysis may gain further understanding from J. Scott Arm¬ strong, “Derivation of theory by means of factor analysis or Tom Swift and his elec¬ tric factor analysis machine”, American Statistician 21:17-21, 1967.

Letter received October 23, 1970

[99]

DR. LESTER WICKS

Dr. Lester Wicks, a member of the Illi¬ nois State Academy of Science and a for¬ mer faculty member of McKendree College passed away in the summer of 1970 after a brain disorder illness of two and one-half years.

Dr. Wicks was born in St. Louis. He received his B.S. and M.S. degrees from St. Louis University and his Ph. D. degree from Washington University in Biochemis¬

try. Before joining the chemistry faculty of McKendree College in 1959, he taught at Parks College from 1955 to 1959. As a research chemist he published extensively in scientific journals. He was a member of Sigma Xi and the Illinois Chemistry Teachers Association. Dr. Boris Musulin, Chemistry, Southern Illinois University, Carhondale, Illinois 62901.

Manuscript received Nov. 29, 1970

[100]

PROFESSOR JOSEPH TYKOCINSKI TYKOCINER

The Illinois State Academy of Science lost a senior member, Professor Emeritus Joseph Tykocinski Tykociner, at the age of 91. Professor Tykociner was referred to as the “father of sound movies”. His first public demonstration of sound on film took place at the University of Illinois in 1922. A permanent display exhibit of his demonstration is in the Ford museum, Greenfield, Michigan. He was born in Vlaclavek, Poland and received his E.E. degree from the Hoheres Tech. Inst. Co- then. He later studied at the Tech. Inst. Berlin and Gottingen and received an hon. Dr. Eng. from Illinois in 1965. Professor Tykociner was a pioneer in wire¬ less transmission, microwaves, and zetetics. He served on the staff of the Marconi Company (England) when the first wireless

transmission was made across the Atlantic ocean. In 1905 he went to Russia to help the Imperial Navy equip itself with radio. He was a research engineer for Westing- house Electric and Manufacturing in 1920 and joined the University of Illinois, de¬ partment of electrical engineering, in 1921. He officially retired in 1948 but at the age of 84 came out of retirement to teach zetetics. Professor Tykociner received the award of merit from the National Electron¬ ics Conference in 1964, one of only three scientists to receive the prize which was instituted twenty years earlier. He was a fellow of the Physical Society. Dr. Boris Musulin, Chemistry, Southern Illinois Uni¬ versity, Carhondale, Illinois, 62901.

Manuscript received October 15, 1970

[101]

DR. BYRON RIEGEL—

PRESIDENT OF AMERICAN CHEMICAL SOCIETY

Dr. Byron Riegel, a member of the Illinois State Academy of Science since 1942, served as the president of the American Chemical Society in 1970. His administration of this society of over 100,- 000 members is another demonstration of the versatility of this outstanding chemist. Dr. Riegel’s academic career was princi¬ pally at Northwestern University from 1937 to 1951 where he attained the rank of professor. His industrial career has been with G. D. Searle & Company as director of chemical research and development since 1951. He also served as a lecturer

with Northwestern from 1951 to 1959. He was born in Palmyra, Mo. and received the A.B. degree from Central Methodist College. His graduate degrees were taken at Princeton and Illinois. He was con¬ ferred the hon. D. Sc. by Central Metho¬ dist College in 1963. Dr. Riegel’s re¬ search has been in Medicinal Chemistry specializing in steroids, structure and bio¬ logical activity, vitamin K, cancer chemo¬ therapy, anti-malarials and other drugs. Dr. Boris Musulin, Chemistry , Southern Illinois University, Carbondale, Illinois 62901.

[102]

BIOLOGICAL NOTES ON PHLOEOTRIBUS SCABRICOLLIS (HOPKINS) (COLEOPTERA : SCOLYTIDAE)

MILTON W. SANDERSON AND JAMES E. APPLEBY Illinois Natural History Survey , Urbana

Abstract. — Phloeotribus scabricollis (Hopkins) is recorded for Illinois and Ohio, the first records since it was original¬ ly described from Hessville, Indiana in 1916. Its occurrence on wafer ash, Ptelea trifoliata — a new host record — is dis¬ cussed.

On July 5, 1970, one of the authors observed insect damage to wafer ash ( Ptelea trifoliata) at the Morton Arboretum, Lisle, Illinois, DuPage County, in northeastern Illinois. Small tunnels and adult scolytid beetles were noted in stems at the bases of the petioles (fig. 1). Further damage and beetles were noted on August 31 and

Figure 1. Base of petiole of wafer ash, Ptelea trifoliata, showing beetles and injury caused by Phloeotribus scabricollis. (Photo by W. Zehr, Illinois Natural History Sur¬ vey).

September 19, but no beetles were in evi¬ dence on October 22, 1970, when the plants were last examined.

The beetle causing the damage was tentatively identified by Sanderson as Phloeotribus scabricollis (Hopkins), and confirmed by Dr. Stephen L. Wood, Brig¬ ham Young University, Provo, Utah, and by Dr. Donald M. Anderson, U.S. National Museum. The species was described by Hopkins (1916, p. 656) in the genus Phloeophthorus from one unassociated spe¬ cimen collected by W. S. Blatchley at Hess¬ ville, Indiana, on July 14; the year of collection not indicated. Hessville now lies within Hammond, Indiana, in extreme northwestern Lake County approximately thirty miles east of Lisle, Illinois. A search of the literature disclosed no further rec¬ ords of this species, but Dr. Wood generous¬ ly gave us an unpublished record of four specimens from Georgesville, Ohio, Franklin County, collected September 18, 1898.

The specimens are labeled as having been collected on bladdernut, Staphylea trifolia. Georgesville, located in the central part of the state, is approximately 250 miles south¬ east of Hessville, Indiana.

Wafer ash and bladdernut are widely distributed in the eastern half of the United States, and both occur in the Morton Arboretum. However, there were no signs of the beetle on bladdernut located about 200 yards from the infestation on wafer ash, and no infestation was found on wafer ash in other areas of the arboretum. Wafer ash and bladdernut examined near Mahom¬ et in Champaign County in east-central Illinois were uninfested. Because of the similarity of Ptelea and Staphylea when not in fruit, it is suggested that Staphylea as a host for Phloeotribus should be confirmed. Hutchinson (1969) shows a close relation¬ ship between the families, Rutaceae and Staphylaceae, to which these genera belong.

Samples of infested Ptelea twigs meas¬ ured from 3 mm to 12 mm in diameter. In one 475-mm long twig, 33 live and 2

[103]

104

Notes

dead adults were found. Usually ther$ was only one beetle in a tunnel in the twig beneath the petiole base, but occasionally two beetles were noted, and in one in¬ stance three betles. Most tunnel entrances were at the base of the petiole, and the tunnel penetrated diagonally to a maximum depth of about 7 mm. However, two en¬ trances were about 3 mm from the petiole base, entering the stem at a right angle and nearly perforating it. Frass usually was present at the entrance of the tunnel, and some tunnels were filled with exudate, completely covering live beetles. These are believed to be feeding tunnels for no larvae were found in them.

The sex ratios of beetles differed marked¬ ly in the three collections as follows : July 5 (9 3, 249); August 31 (9 3, 89); September 19 (23, 59).

Literature Cited

Hopkins, A. D. 1916. In Rhynchophora or Weevils of North Eastern America by W. S. Blachley and C. W. Leng. The Nature Publishing Co., Indianapolis.

682 pp.

Hutchinson, J. 1969. Evolution and Phylogeny of Flowering Plants. Aca¬ demic Press, New York. 717 pp.

Manuscript received Nov. 20, 1970

PREPARATION OF MANUSCRIPTS FOR THE TRANSACTIONS

For publication in the Transactions, articles must present significant material that has not been published elsewhere. Review articles are ex¬ cepted from this provision, as are brief quotations necessary to consider new material or varying concepts. All manuscripts piust be typewritten, double spaced, with at least one-inch margins. The original copy and one carbon copy should be submitted.

Titles should be brief and informative. The address or institutional connection of the author appears just below the author’s name. An abstract must accompany each article. Subtitles or center headings should be used; ordinarily one uses subtitles such as Materials and Methods, Results, Dis¬ cussion, Summary, Acknowledgments, and Literature Cited.

No footnotes are to be used except in tables.

The section entitled Literature Cited must include all references men¬ tioned in text. It is not to include any other titles. Citations under Literature Cited are as shown below:

Doe, J. H. 1951. The life cycle of a land snail. Conchol., 26(3): 21-32, 2 tables, 3 figs.

Doe, J. H., and S. H. Jones. 1951. Mineralogy of Lower Tertiary deposits. McGraw-Hill Book Co., New York, iv + 396 pp.

Quoted passages, titles, and citations must be checked and rechecked for accuracy. Citations to particular pages in text are Doe (1908, p. 21) or (Doe, 1908, p. 21) ; general citation in text is Doe (1908) or (Doe, 1908).

Tabular information should be kept at a minimum. Do not duplicate tabular data in text. Headings for tables and columns should be brief. Each table and its heading should be on a single page; do not place any table on the same page with text.

Photographs should be hard, glossy prints of good contrast. Graphs, maps and other figures reproduce best when prepared for at least one-half reduction; lettering, numerals, etc. on all figures in a manuscript should be worked out to proper size for such reduction. Line widths, letter size, etc., should be uniform from figure to figure within a published paper. Figures should be drawn on good quality white paper or on drawing boards. Drawings should be no more than 10" x 14", preferably Sy2" x 11". Use only India ink. Use a lettering device (Leroy or Wrico) for numerals and words; do not print “free-hand.”

Legends for figures should be brief; type them all on a single sheet of paper. Indicate figure number and your name on back of illustration; do not write with pencil on the backs of photographs.

Authors will receive galley proofs; these should be read carefully and checked against the original manuscript. The editor reserves the right to make last minute corrections (spelling, punctuation generally) without consultation with the author, and to alter suggested running titles. Re¬ prints may be ordered at the time galley proofs and manuscripts are re¬ turned to the Editor.

Malcolm T. Jollie,

Department of Biological Sciences, Northern Illinois University, DeKalb, Illinois 60115

Transactions

of the

Illinois

NEW

V

&

} K?

IK

BOTANICAL GARDEN

State Academy of Science

Volume 64 No. 2 1971

TISAAH

PREPARATION OF MANUSCRIPTS FOR THE TRANSACTIONS

For publication in the Transactions , articles must present significant material that has not been published elsewhere. Review articles are ex¬ cepted from this provision, as are brief quotations necessary to consider new material or varying concepts. All manuscripts must be typewritten, double spaced, with at least one-inch margins. The original copy and one other copy should be submitted.

Titles should be brief and informative. The address or institutional connection of the author appears just below the author’s name. An abstract must accompany each article. Subtitles or center headings should be used; ordinarily one uses subtitles such as Materials and Methods , Results, Dis¬ cussion, Summary, Acknowledgments, and Literature Cited.

No footnotes are to be used except in tables.

The section entitled Literature Cited must include all references men¬ tioned in text. It is not to include any other titles. Citations under Literature Cited are as shown below:

Doe, J. H. 1951. The life cycle of a land snail. Conchol., 26: 21-32, 2 tables, 3 figs.

Doe, J. H., and S. H. Jones. 1951. Mineralogy of Lower Tertiary deposits. McGraw-Hill Book Co., New York, iv + 396 pp.

Quoted passages, titles, and citations must be checked and rechecked for accuracy. Citations to particular pages in text are Doe (1908, p. 21) or (Doe, 1908, p. 21) ; general citation in text is Doe (1908) or (Doe, 1908).

Tabular information should be kept at a minimum. Do not duplicate tabular data in text. Headings for tables and columns should be brief. Each table and its heading should be on a single page; do not place any table on the same page with text.

Photographs should be hard, glossy prints of good contrast. Graphs, maps and other figures reproduce best when prepared for at least one-half reduction; lettering, numerals, etc. on all figures in a manuscript should be worked out to proper size for such reduction. Line widths, letter size, etc., should be uniform from figure to figure within a published paper. Figures should be drawn on good quality white paper or on drawing boards. Drawings should be no more than 10" x 14", preferably 8%" x 11" (or 8" x 10"). Use only India ink. Use a lettering device (Leroy or Wrico) for numerals and words; do not print “free-hand.”

Legends for figures should be brief; type them all on a single sheet of paper. Indicate figure number and your name on back of illustration in blue pencil.

Authors will receive galley proofs; these should be read carefully and checked against the original manuscript. The editor reserves the right to make last minute corrections (spelling, punctuation generally) without consultation with the author, and to alter suggested running titles. Re¬ prints may be ordered at the time galley proofs and manuscripts are re¬ turned to the Editor.

Malcolm T. Jollie,

Department of Biological Sciences, Northern Illinois University, DeKalb, Illinois 60115

TRANSACTIONS

OF THE

ILLINOIS STATE ACADEMY OF SCIENCE

VOLUME 64 - 1971

No. 2

Illinois State Academy of Science

AFFILIATED WITH THE

Illinois State Museum Division Springfield, Illinois

PRINTED BY AUTHORITY OF THE STATE OF ILLINOIS Richard B. Ogilvie, Governor

June 1, 1971

CONTENTS

Excess Molar Volumes of Mixing of Solutions of Nitromethane and Carbon Tetrachloride

By John F. Wettaw and Boris Musulin . 107

The Evolution of Growth Habit in Cynodon L. C. Rich ( Gramineae )

By Kantilal M. Rawal and Jack R. Harlan . 110

Derivation of 1/Z Expansion of Hartree-Fock Equations

By Yuh Kang Pan . 119

Death of Cells in Pith Tissue of Soybean Seedlings

By Kuo-Chun Liu and A. J. Pappelis . 128

Microbodies of Soybean Cotyledon Mesophyll

By Kuo-Chun Liu, A. J. Pappelis, and H. M. Kaplan . 136

Thin Films of Interphase Chromatin Prepared for Electron Microscopy by Osmotic Shock Technique

By Fathi Abdel-Hameed . 142

Pathogenesis of Clostridium Botulinum: In Vivo Fate of C. Botulinum Type A Spores

By R. Booth, J. B. Suzuki and N. Grecz . 147

Three Iron Sulfate Minerals From Coal Mine Refuse Dumps in Perry County,

Illinois

By Dennis Gruner and William C. Hood . 156

The Behavior of Iron in Peoria Lake

By Wun-Cheng Wang and Ralph L. Evans . 159

Improved X-Radiography of Cylindrical Sediment Cores

By Gordon S. Fraser and Adrain F. Richards . 169

The Spatial Distribution of Lake-Effect Snowfall within the Vicinity of Lake Michigan

By Kenneth Frederic Dewey . 177

The Coupling of Energy Production to Synthesis in the Original Operation of Living Systems

By Newton Ressler . 188

Notes

Gideon Herman Boewe, 1895-1970

By Robert A. Evers and J. Cedric Carter . 193

A Leucistic Little Brown Bat ( Myotis L. Lucifugus )

By Harlan D. Walley . 196

The First Record in Illinois of a Population of Stethaulax Marmoratus (Say)

(Hemipetera: Scutelleridae) with Information on Life History J. E. McPherson and J. F. Walt . 198

Mr. Stover

By Hiram F. Thut and John Ebinger . 201

Contributing Members of 1970 . 203

EXCESS MOLAR VOLUMES OF MIXING OF SOLUTIONS OF NITROMETHANE AND CARBON TETRACHLORIDE

JOHN F. WETTAW AND BORIS MUSULIN Department of Chemistry ,

Southern Illinois University, Carbondale, Illinois 62901

Abstract. — The direct measurement, by means of weight and density measurements, of the excess molar volume of mixing in solutions of nitromethane and carbon tet¬ rachloride at 35 °C. is described. The data confirm the quadratic nature of deviations from ideality for these solutions.

Gunter et al. (1967) have reported values of excess molar volumes of mixing' of binary solutions of CH3N02 and CC14 which were de¬ rived from measured densities as¬ suming that the molar volumes of mixing of ideal solutions were addi¬ tive. Although the reported values are consistent in functional form and magnitude with those reported by Brown and Smith (1955), they appear to be of lesser quality. The purpose of this investigation is to ascertain if directly measured values, from a simple experiment, are of better quality than derived values.

Experimental

Fisher Spectrograde nitromethane and carbon tetrachloride were used without further purification. Tem¬ perature, specific gravity, and weight measurements were made as de¬ scribed in Gunter et al. (1967). A temperature of 35 °C. was used in this work. Two sets of pipets, one

for CH3N02 and one for CC14, each containing a 25-ml, a 10-ml, and a 5-ml pipet, were calibrated by weight techniques. Of the seven physically independent permutations that can be made by mixing a pipet- ful from each set, four were used to make the sample solutions whose weights were determined directly. The solution densities were meas¬ ured directly and the solution vol¬ umes calculated from these densities and weights. The densities of the pure components used in calibration were taken from Gunter et al. (1967). The excess molar volumes of mixing were obtained bv differ- ences of the measured volumes.

Results and Discussion

All measured and calculated val¬ ues are given in Table 1. The mole fraction values for each solution are at least one magnitude more reliable than those given by Gunter et al. (1967). In calculations, the mole fraction was used, as reported, with four significant figures.

The measured solution densities are compared to values calculated from the density -mole fraction func¬ tion derived by Musulin (1971).

(107]

108

Transactions Illinois Academy of Science

Table 1. — Densities and Molar Volumes of Mixing of Nitromethane-Carbon

Tetrachloride Solutions

Mole

Fraction

(CH3NO2)

Density (g/ m0

Calculated

Densitya

(g/ml)

Molar Volume of Mixing (ml/mole)

Calculated Molar Volume of Mixing3 (ml/ mole)

Additive Molar Volume of Mixing (ml/ mole)

0.4174

1.43137

1.43479

0.2064

0.1990

0.2404

0.6414

1.33347

1.33201

0.2088

0.1925

0.2465

0.8178

1.23861

1.23410

0.1078

0.1189

0.1652

0.8989

1.18568

1.18408

0.06250

0.07192

0.04940

a. Musulin ( 1971 )

The maximum difference is 0.0045 g/ml, i.e. 0.36%. Excluding- pure experimental error, two reasons may be given for the variation. First, Musulin derived the function using lesser quality mole fraction data, e. g. the 0.8989 mole fraction CH.r X02 solution of the present work would have been reported as 0.900 mole fraction CH3N02 in the earlier work. Second, the temperature vari¬ ation (inherent in the Musulin func¬ tion) which was allowable with lesser quality mole fraction data becomes a controlling factor in the present investigation. Nevertheless, it is clear that the present densities are in good agreement with the earlier work.

The measured excess molar vol¬ umes of mixing are compared to val¬ ues calculated from the excess molar volume of mixing-mole fraction function derived by Musulin (1971) and to values calculated assuming additivity of molar volumes for ideal solutions as was done by Gunter et al. (1967). The measured values are in good agreement with those cal¬ culated by the Musulin function. The deviations were about 0.01 ml.

As before, these variations could be attributed to mole fraction and tem¬ perature measurements. The in¬ creased magnitude of the variation could be attributed to the fact that excess molar volume of mixing is a difference quantity. Comparison to the values obtained from the ad¬ ditivity assumption indicates that values obtained by that assumption are overstated in the middle of the mole fraction range. The existence of this error introduces another er¬ ror in the function derived by Musu¬ lin and establishes the final reason for the greater deviations from cal¬ culated values with excess molar volumes of mixing compared to the deviations with densities.

These new, accurate measure¬ ments confirm the quadratic form and the magnitude of the deviation from ideality of solutions of CH3N02 and CC14. The weight techniques presented in this work provide a meaningful way to obtain better quality data than that obtained by Gunter et al. (1967) and is an ap¬ propriate compromise if the vapor density equipment is not available. Finally, the present results empha-

Wettaw and Masulin

Excess Molar Volumes

109

size the necessity of utilizing an ex¬ cess type equation for the excess molar volumes of mixing for CH3- N02-CC14 solutions as opposed to utilization of a simple additive as¬ sumption.

Acknowledgment

The authors gratefully acknowledge the support of the Petroleum Research Fund (Grant #602-B) Administered by the American Chemical Society for Support during the course of this work. They are also indebted to Prof. K. A. Van Lente for assistance with the experimental tech¬ niques.

Literature Cited

Brown, I. and F. Smith. 1955. Liquid Vapour Equilibria. VII. The Systems Nitromethane and Benzene and Nitro- methane and Carbon Tetrachloride at 45°C. Australian J. Chem. S; 501-505.

Gunter, C. R., J. F. Wettaw, J. D. Dren- nan, R. L. Motley, M. L. Coale, T. E. Hanson, and B. Musulin. 1967. Den¬ sities and Molar Volumes of Binary Solu¬ tions of Nitroparaffins in Carbon Tetra¬ chloride. J. Chem. Eng. Data 12: 472- 474.

Musulin, B. 1971. Computerized Curve Fitting: An Alternative to Graphical

Interpretation. Trans. Ill. State Acad. Sci. 64: 67-83.

Manuscript received October 19, 1970.

THE EVOLUTION OF GROWTH HABIT IN CYNODON

L. C. RICH. (GRAMINEAE)

KANTILAL M. RAWAL AND JACK R. HARLAN

Agronomy Department, University at Illinois, Urbana 61801

Abstract. — Vegetative characters stud¬ ied included presence or absence of rhi¬ zomes, kind of rhizome, modification of stolon tips, branching patterns and winter hardiness. Growth habit, together with geographic distribution, ecological adapta¬ tion, and affinity based on cytogenetic studies suggests a sequence of evolutionary events in Cynodon.

A biosystematic study of the genus Cynodon was conducted at Stillwater, Oklahoma from 1963 to 1967 result¬ ing in a revision of the genus (Clay¬ ton and Harlan 1970, Harlan and de Wet 1969, Harlan et al. 1969). The species and varieties as now recognized are presented in Table 1.

Table 1. — The Species and Varieties of Cynodon.

Epithet

2n

Chromosome

Number

Distribution

C. aethiopicus Clayton et Harlan .

18, 36

East Africa; Ethiopia to Transvaal

C. arcuatus J. S. Presl ex.

C. B. Presl .

36

Malagasy, Ceylon, India, Southeast Asia, Philippines, Taiwan,

Indonesia to Australia

C. barberi Rang, et Tad .

18

South India

C. dactylon (L.) Pers

var. dactylon .

36

Cosmopolitan

var. afghanicus Harlan et de Wet .

18, 36

Afghanistan

var. aridus Harlan et de Wet .

18

South Africa to Palestine to

South India; intro, in Hawaii, Arizona

var. coursii (A. Camus)

Harlan et de Wet .

36

Madagascar

var. elegans Rendle .

36

Southern Africa; Mozambique, Zambia and Angola southward

var. polevansii (Stent) Harlan

et de Wet .

36

Baberspan, South Africa

C. incompletus Nees

var. incompletus .

18

South Africa

var. hirsutus (Stent) Harlan

et de Wet .

18, rarely 36

South Africa

C. nlemfuensis Vanderyst

var. nlemfuensis .

18, rarely 36

Tropical Africa; Ethiopia to

Zambia, west to Angola

var. robustus Clayton et Harlan .

18, 36

East Africa; Ethiopia to Rhodesia

C. plectostachyus (K. Schum.)

Pilger .

18

Ethiopia, Uganda, Kenya,

Tanzania

C. transvaalensis Burtt-Davy .

18

Transvaal and Orange Free State

[110]

Rawal and Harlan — Evolution in Cynodon

111

Specimens were examined at Kew, British Museum, Paris, Brussels, Berlin, Florence, Geneva, Honolulu, Bangkok, Manila, Los Banos, and Washington. The revised classifica¬ tion, however, was based primarily on a living collection of some 700 accessions grown in uniform nurser¬ ies at Stillwater. In the living ma¬ terial, it was noted that most of the taxa could be distinguished bv characteristic growth habits. Vege¬ tative characters are difficult to de¬ scribe and are highly subject to en¬ vironmental modification, yet they are useful for field identification and have evolutionary implications. They tend to go unnoticed in the herbari¬ um because few specimens are suf¬ ficiently complete to reveal growth characteristics. The main types of growth habits are described in this paper as an aid to identification and to shed some light on evolution in the genus.

Description

RHIZOMES : The genus is essen¬ tially caespitose ; only C. transva- alensis and four of the six varieties of C. dactylon have rhizomes. There are, however, two kinds of rhizomes, Fig. 1. One tends to be relatively slender, straight, with long inter¬ nodes and the tip always stays below the surface. Lateral buds grow up¬ ward and emerge to form culms, but the rhizome itself remains under¬ ground, Fig. 1A. The other kind of rhizome is relatively large in diame¬ ter, fleshy, usually crooked with short internodes and the tip may grow to the surface where the rhi¬ zome is converted into a stolon, Fig. IB. The first type grows faster and

deeper than the second. The dis¬ tribution of rhizome types is given in Table 2.

always stays below the soil surface.

B. Rhizome that emerges and is con¬ verted to a stolon.

The inheritance of rhizome forma¬ tion was studied in a limited number of the hybrids produced in the course of the biosystematic study. Table 3. In hybrids between non- rhizomatous and rhizomatous species, rhizome formation was suppressed, but the nonrhizomatous varieties of C. dactylon were not able to suppress rhizome formation. The tetraploid race of C. nlemfuensis var. robustus does not suppress rhizomes complete¬ ly, and, indeed, rhizomes were pro¬ duced in hybrids between it and

112

Transactions Illinois Academy of Science

Table 2. — Some Characteristics of Growth Habit in Cvnodon.

Taxon

Rhizome

Type

(Fig. 1)

Stolon

Tip

(Fig. 2)

Stolon Branching (Fig. 4)

Turf

Formation

Size*

Tissue

Hardiness

C. aethiopicus .

None

A

A

Very Open

Large

None

C. arcuatus .

None

A

A

Open

Small

None

C. barberi .

None

A

A

Open

Small

None

C. dactylon

var. dactylon .

A & B

A, R, B

B, C

Dense

Small-

Medium

V ariable

var. afghanicus .

None

A

A

Very Open

Medium

Yes

var. aridus .

A

A

A

Open

Small-

Medium

None

var. coursii .

None

R

B

Dense

Large

None

var. elegans .

B

B

A

Lax

Medium

None

var. polevansii .

A

B

C

Dense

Small

Yes

C. incompletus

Yes

var. incompletus . . . .

None

R

B

Dense

Small

var. hirsutus .

None

R

B

Dense

Small

Yes

C. nleriifuensis

var. nlemf uensis . . . .

None

A

A

Open

Medium-

Large

None

var. robustus (lx) . . .

None

A

A

Very Open

Large

None

var. robustus (4x) . . .

None

A

C

Open

Large

None

C. plectostachyus .

None

A

B

Open

Large

None

C. transvaalensis .

B

B

B

Dense

Small

Yes

* Small plants are usually less than 15 cm tall; large plants are usually over 40 cm tall under nursery conditions.

Table 3. — Rhizome Formation in Hybrids Between Nonrhizomatous and

Rhizomatous Taxa

Nonrhizomatous Parent

C. aethiopucus (4x) .

C. incompletus (lx) .

C. incompletus (2x) .

C. nlemfuensis var. nlemfuensis (2x) C. nlemfuensis var. nlemfuensis (2x) C. nlemf uensis var. robustus (2x) . . . C. nlemfuensis var. robustus (4x) . . . C. dactylon var. afghanicus (2x)

C. dactylon var. afghanicus (2x, 4x) .

C. dactylon var. coursii (4x) .

C. dactylon var. coursii (4x) .

C. dactylon var. coursii (4x) .

Rhizomatous Parent

C. dactylon (4v) .

C. dactylon (fix) .

C. dactylon (4ar) .

C. dactylon ( lx ) .

C. dactylon (4v) .

C. dactylon (lx) .

C. dactylon (4v) .

C. dactylon (lx) .

C. dactylon (4v) .

C. dactylon (lx) .

C. dactylon (4v) .

C. transvaalensis (lx)

Number

Hybrids

Examined

Rhizome

Formation

4

3

—

65

—

35

—

22

—

4

—

15 + 4*

—

35

+

47

+

41

+

43

6

+

* Four Fx plants of this combination had short, poorly developed rhizome-like structures.

Raival and Harlan — Evolution in Cynodon

113

tetraploid C. dactylon var. afghani- cus. The fact that nonrliizomatous parents could produce rhizomatous offspring* suggests that both parents carried recessive genes for rhizome production. Our collection also con¬ tained populations of C. dactylon var. coursii that apparently had al¬ ready crossed with var. dactylon in Madagascar and segregated for rhi¬ zome production in our nurseries. The character gives every evidence of being rather simply inherited. STOLON TIPS : There are strik¬ ing differences among taxa in the morphology and behavior of stolon tips. Two extremes are shown in Fig. 2. In one kind, the tips are soft and leafy and the blades, when fully developed, are little different from those of the culms, Fig. 2A. In the other type, the blades are re¬ duced to small flaps and the encasing sheaths are hardened, producing a sharp organ capable of penetrating soil, Fig. 2B. The organ resembles the penetrating tip of a rhizome, and, in fact, is readily converted into a rhizome when buried in the soil. Stolons with leafy tips simply stop growing when buried.

In some races of C. dactylon var. dactylon, the stolons regularly bury themselves, pushing the sharp tips down into the soil where the stem is converted to a rhizome which may emerge again after a short distance to be reconverted into a stolon. The horizontal stems of these plants seem to “ gallop” alternately plung¬ ing into the soil and emerging again, producing rhizomes and stolons al¬ ternately along the length of the stem.

The extremes, as figured, are easily recognized, but there are intermedi¬

ate forms that are very difficult to classify, especially with herbarium specimens. In C. incompletus , for example, the blades of the stolon leaves near the tip are much reduced and they never develop as fully as the leaves on the culm. The stolon tip may resemble the sharp-pointed organ just described, but it is softer and cannot be converted into a rhi¬ zome by burial in the soil. In Table 2, we have listed such intermediates as “R” for the reduced blades of stolon leaves. C. dactylon var. dac- tylon is especially variable and all

Figure 2. Stolon tips.

A. Leafy type relatively unmodified.

B. Extreme of modified type in which the sheaths are hardened and the blades much reduced.

114

Transactions Illinois Academy of Science

classes of stolon tips are found in it.

BRANCHING HABIT : Branching- patterns depend on the number and position of buds differentiated and on the timing of their development. In most grasses, leaves are arranged alternately in two ranks on the culm, one leaf to the node and each sub¬ tending a lateral bud in the axil. In Cynoclon and some other grasses the culms have two leaves at each node, but only the lower subtends a lateral bud (Bogdan 1952). If the nodes were truly compound, one would ex¬ pect each leaf to subtend a bud. The culm nodes could be interpreted as simple with the upper leaf subtend¬ ing the terminal bud even though the terminal bud becomes far re¬ moved by subsequent growth of the culm, Fig. 3A. The lateral bud is usually suppressed, although those at the base of the culm may develop into a branch if the culm is allowed to develop fully.

The stolon nodes of Cynodon are compound and have three leaves, the lower two subtending lateral buds on opposite sides of the stolon, Fig. 3B. In some taxa, notably races of C. dactylon var. dactylon, the stolon nodes appear to have more than three leaves, but careful exami¬ nation of young nodes near the tip shows that the basic ground plan is essentially constant. The addi¬ tional leaves are derived from pre¬ cocious growth of the lateral buds.

Of the two buds, the lower one is the first to grow and develop into either a stolon branch or a culm. Growth of the upper bud may be suppressed for a long time resulting in a stolon with conspicuously alter¬ nate branching, Fig. 4A. In such

Figure 3. Diagram of culm leaves, A and stolon leaves, B.

Figure 4. Branching patterns.

A. Alternate, one bud suppressed.

B. Intermediate, one bud delayed.

C. Opposite subequal.

Rawal and Harlan — Evolution in Cynodon

115

stolons, rooting is usually also sup¬ pressed so that in the larger forms, one may lift up a stolon one or two meters long unattached to the soil, and with regular alternate branches down its length. Under natural con¬ ditions, stolons of this type may fes¬ toon shrubs and even small trees three meters or more in height. They tend to have long internodes ( > 10 cm and sometimes > 20 cm) and are very fast growing. Under nurs¬ ery conditions they may grow 10 meters or more in a single season. Plants of this type produce a loose, open mat of growth rather than a dense turf.

At the other extreme, are taxa in which the upper bud at a node de¬ velops almost immediately after the lower one, producing a branching pattern that appears to be opposite and subequal, Fig. 4C. These tend to have short internodes, root freely, and form a dense turf covering the soil almost completely. Further¬ more, the branches themselves branch quickly and nodes only a short dis¬ tance back from the tip may show a knot of growth consisting of sev¬ eral culms, short stolon branches, and many leaves. In the absence of competition, the turf creeps in a closed, dense front across the soil surface, rooting immediately behind the stolon tips.

Again, the two extremes are con¬ spicuously different, but some taxa show intermediate behavior as in Fig. 4B. An individual plant may also be rather variable. During periods of maximum growth, inter¬ node elongation is sufficiently rapid that the branching appears alternate, but as growth slows down the de¬ velopment of the two buds at a node

become more nearly synchronous and the branching approaches opposite. The upper lateral bud usually grows eventually even in forms that have a distinctly alternate branching pat¬ tern. Despite the variability, branch¬ ing habits of some taxa are con¬ spicuously different from others. Cynodon aethiopicus, C. nlemfuensis

var. nlemfuensis and 2x var. robus-

tus, C. arcuatus, C. barberi and C. dactylon var. aridus, afghanicus and elegans have consistently alternate branching, at least along the distal portions of the stolons. Cynodon plectostachyus, C. nlemfuensis var. robustus (4x), C. incompletus, C. transvaalensis, and C. dactylon vars. dactylon , coursii, and polevansii have essentially opposite branching pat¬ terns.

SIZE : There are striking differ¬ ences among taxa with respect to plant size. This, again, is a variable character and readily influenced by environment. Under conditions of a uniform nursery in full sunlight and with competition with other vegetation removed, the differences are so consistent that plant size is one of the most conspicuous of all characters. Under these conditions, small plants are usually 15 cm or less in height and large plants are generally 40 cm or more. Some of the most robust forms may exceed a meter in height. A rough classi¬ fication of plant size is presented in Table 2.

HARDINESS : Cynodon is basical¬ ly a tropical genus and plants of most species are very sensitive to freezing. Plants of C. arcuatus, C. barberi, C. aethiopicus, C. plecto¬ stachyus, C. nlemfuensis, C. dacty¬ lon var. coursii and some races of

116

Transactions Illinois Academy of Science

var. dactylon are completely de¬ stroyed by a killing’ frost under Ok¬ lahoma conditions. C. dactylon var. arid us apparently has no tissue hardiness, but can usually overwin¬ ter at Stillwater by virtue of the deep rhizomes that escape killing temperatures. C. dactylon var. ele- gans has the same faculty, but the rhizomes do not go as deep and mor¬ tality is very high. Plants of the tropical race of var. dactylon may survive especially mild winters, but always with very severe injury. Deep rhizomes can act as a survival mechanism for plants without tissue hardiness.

Cynodon incompletus and C. dac¬ tylon var. afghanicus, on the other hand, have good tissue hardiness but no rhizomes. Both overwinter well in Oklahoma. Cynodon transvaalen- sis, C. dactylon var. polevansii and many accessions of var. dactylon have both tissue hardiness and rhi¬ zomes.

LEAF SHAPE : Three species can be easily recognized by leaf shape. In C. barbeid, the leaves are broadly ovate-lanceolate, conspicuously dif¬ ferent from all other taxa in the genus. In C. arcuatus, the leaves are broadly linear-lanceolate, rather intermediate between C. barberi and most of the other taxa. There is no overlap, however, and the leaves of C. arcuatus are readily recogniz¬ able. Cynodon transvaalensis repre¬ sents the other extreme, with slender linear leaves finer than in any other species in the genus. Plants of all other taxa have linear-lanceolate leaves more or less alike in form. There is a conspicuous range in size, but only the three species mentioned

can be consistently distinguished by leaf shape.

Interpretation

Cynodon barberi and C. arcuatus are well separated from the rest of the genus not only by leaf shape and growth habit, but by inflores¬ cence and spikelet characters and by genetic barriers (Harlan and de Wet 1969, Harlan et al. 1969). The distribution of C. arcuatus across the islands of the Indian and South Pa¬ cific Oceans from the Comoros and Seychelles to Australia suggests an ancient distribution of ancestral forms, clearly distinct from the geo¬ graphic patterns of the rest of the genus. Cynodon barberi shows some morphological affinity to the nearest genus, Brachyachne, which is rep¬ resented by a number of species in both tropical Africa and Australia that were at one time assigned to Cynodon. Cynodon barberi and C. arcuatus appear, therefore, to rep¬ resent a very early differentiation from the ancestral Cynodon stock and are no longer closely related to the remaining taxa.

A second clearly separable group includes the large East African spe¬ cies C. aethiopicus, C. nlemfuensis, and C. plectostachyus. They share a number of growth habit charac¬ teristics such as lack of rhizomes, leafy stolon tips, lack of hardiness, large size, open growth and mostly alternate branching patterns. All three have distributions closely as¬ sociated with the Great Rift Valley. In crossability studies reported by Harlan et al. (1969), it was shown that C. plectostachyus is completely

Rawal and Harlan — Evolution in Cynodon

117

isolated genetically from other taxa and that C. aethiopicus is isolated by very strong- genetic barriers.

A third natural group includes the South African endemics, C. in- completus and C. transvaaleyisis. They are small, turf-forming di¬ ploids with far more winter hardi¬ ness than is required by their pres¬ ent habitats in South Africa. Al¬ though sympatric in part, they do not seem to cross in nature, but can be hybridized artifically (Harlan et al. 1970).

In the complex species C. dacty- lon, there is nothing that appears directly related to the first group. C. dactylon var. coursii, however, is a large, tropical, nonhardy and nonrhizomatous form with evident connections to C. nlemfuensis. Har¬ lan et al. (1969) report that it crosses rather easily with other varie¬ ties of C. dactylon and that some hy¬ brids with C. nlemfuensis were pro¬ duced. The variety polevansii is a small, turf -forming, winterhardy en¬ demic of South Africa evidently as¬ sociated with the third group. The other African variety, elegans has growth habits that suggest it is a tetraploid form derived, in part at least, from the diploid var. aridus.

Finally, the Asian forms of C. dactylon seem to form a fourth group. Harlan and de Wet (1969) have shown that var. aridus, var. af ghanicus, and the winterhardy temperate races of var. dactylon in¬ teract genetically in Asia. Intro- gression between hardy races of var. dactylon and hardy var. af ghanicus is especially evident in Afghanistan. There is no apparent genetic connec¬ tion between the winterhardy forms

of Asia and those of South Africa. Hardiness has evolved independently in two widely separated regions. Furthermore, it is the Asian group that has produced the cosmopolitan weed var. dactylon (Harlan and de Wet 1969).

We, therefore, postulate the fol¬ lowing sequence of evolutionary events :

1) . C. barheri and C. arcuatus separated early from the remainder of the Cynodon stock. They are dif¬ ferent morphologically, completely isolated genetically, and have dis¬ tributions and affinities that suggest an early differentiation. Croizat (1968) assigns distributions of this type to precretaceous events.

2) . The large, robust forms of East Africa (C. aethiopicus, C. nlemfuensis, C. pie dost achy us) evolved, adapted to rather high rain¬ fall and warm temperatures. Their distributions are closely associated with the Great Rift Valley and ad¬ jacent highlands which began to as¬ sume their present conformation in mid-Tertiary.

3) . A diploid form evolved rhi¬ zomes and invaded the more arid re¬ gions of Africa, the Near East, and India. There were both small and large races and C. dactylon var. ari¬ dus represents the modern survivor of the progenitor rhizomatous C. dactylon.

4) . The South African endemics evolved in isolation from the rhi¬ zomatous forms of India and the Near East. Winterhardy forms, with and without rhizomes, emerged more or less simultaneously in South Africa ( C . incompletus, C. transvaal- ensis) and in Asia (C. dactylon

118

Transactions Illinois Academy of Science

var. afghanicus and var. dactylon) . The high degree of winterhardiness in the South African species implies natural selection and survival through the Pleistocene.

5). C. dactylon var. dactylon, in part through genetic interaction with var. afghanicus and var. aridus be¬ came a cosmopolitan weed (Harlan and de Wet 1969). The high degree of winterliardiness of the temperate races of this variety also implies a Pleistocene evolution, but the cos¬ mopolitan distribution, especially in the Pacific Islands and the New World is a recent historical phe¬ nomenon.

Summary

Although highly subject to en¬ vironmental modification, the vege¬ tative characters in the genus Cyno- don are useful for field identification and have evolutionary implications. Detailed observations of growth hab¬ it and characteristic features of rhi¬ zomes and stolons are described.

The genus is essentially nonrhi- zomatous ; only Cynodon transvaal- ensis and four of the six varieties of C. dactylon have them. In some taxa, the rhizomes and stolons are interconvertible ; others have true rhizomes and true stolons that are not convertible.

The compound nodes of stolons have three leaves, each subtending

a bud, one terminal and two lateral. Of the two lateral buds, the lower one grows first and develops into either a stolon or a branch. The second bud grows after a delay of varying duration. The length of the delay accounts for the branch¬ ing patterns of stolons.

A sequence of evolutionary events in Cynodon has been postulated after taking into account the information available from geographic distribu¬ tion and cytogenetic studies.

Acknowledgment

The work was supported in part by Na¬ tional Science Foundation Grants GB201 and GB2686.

Literature Cited

Bogdan, A. V. 1952. Observations on stoloniferous grasses in Kenya. J. East Afr. Nat. Hist. Soc. 20: 71-76. Clayton, W. D. and J. R. Harlan. 1970. The genus Cynodon L. C. Rich, in Tropical Africa. Kew Bull. 24: 185-189. Croizat, Leon. 1968. Introducion rai- sonee a la biogeographie de l’Afrique. Mem. Soc. Brot. Vol. 22, Coimbra. Harlan, J. R. and J. M. J. de Wet. 1969. Sources of variation in Cynodon dactylon (L.) Pers. Crop Sci. 9: 774-778. Richardson, W. L. 1969. Hybridization studies with species of Cynodon from East Africa and Malagasy. Amer. J. Bot. 56: 944-950.

- , M. R. Felder, K. M.

Rawal and W. L. Richardson. 1970. Cytogenetic studies in Cynodon L. C. Rich, (Gramineae). Crop Sci. 10: 288-291.

Manuscript received May 21, 1970

DERIVATION OF 1/Z EXPANSION OF HARTREE-FOCK

EQUATIONS

YUH KANG PAN Department of Chemistry, Boston College,

Chestnut Hill, Massachusetts 02167

Abstract. — A simple method of deriving the 1/Z expansion of the Hartree-Fock equation by using the properties of determinants and the orthonormalities of spin and space is described. The method consists of an elementary derivation of the energy ex¬ pression and subsequent variation of it using Lagrange multipliers. It can also be used to derive the regular (restricted or unrestricted) Hartree-Fock equations.

In recent years, the usefulness of the 1/Z expansion perturbation method for calculating atomic wave functions and energy states, and for predicting atomic properties has been widely explored (Hirschf elder, Brown and Epstein, 1964; Dalgarno, 1962). The method involves use of the perturbation parameter, which is the inverse nuclear charge Z. It has the advantage that all members of an iso-electronic sequence may be treated simultaneously. Furthermore, it also provides a convenient basis for com¬ parison with similar expansions corresponding to the non-relativistic many- electron Schrodinger equation. The zeroth-order equations of this method are satisfied by hydrogenic wave-functions and energies ; the first-order perturbed equations using this method have been solved for a number of systems either variationally (Dalgarno, 1960) or in terms of infinite sums of Laguerre functions) Linderberg, 1961 ; Sharma and Coulson, 1962). For predicting many atomic properties, the calculations are very simple using this method, convergence is rapid and results are comparable in accuracy with those of the Hartree-Fock approximation. The 1/Z expan¬ sion method may be developed further to yield results superior in accuracy to those of the Hartree-Fock approximation. It is well known that the Hartree-Fock approximation has played an important role in the develop¬ ment of theories of atomic structure. Its quantitative application leads to a set of coupled integro differential equations, specific to the particular atomic system under consideration, which can be solved only by lengthy numerical techniques. If we apply the 1/Z expansion method to the Hartree-Fock equations, it leads to sets of uncoupled ordinary differen¬ tial equations, specific to the particular electron shell under consideration, which can be solved exactly in analytical form. For this reason, many calculations have been done by this method in the last few years (e.g. Cohen and Dalgarno, 1961, 1963a, 1963b, 1964, 1965a, 1965b, 1967 ; Schwartz, 1962; Coulson and Hibbert, 1967; Stewart, 1964; Sharma, 1962). How¬ ever, no detailed derivation of the 1/Z expansion of the Hartree-Fock

[119]

120

Transactions Illinois Academy of Science

equations has been shown in any of those papers or in any standard quan¬ tum mechanics textbooks except perhaps for a very brief description of the standard procedure of the derivation. The purpose of the present note is to show that a simple method of derivation can be obtained by using the properties of determinants, spin orthonormality and space orthonor¬ mal it v.

Notation

In order to illustrate the procedure, let us consider the case of the restricted Hartree-Fock wave function for a ls22p22p state of excited lithium atom by using the following notations :

^(i) : Is orbital with a spin for the ith electron.

S(i) : Is orbital with B spin for the ith electron.

£0 (i) : 2p0 orbital with a spin for the ith electron.

PQ(i): 2pe orbital with B spin for the ith electron.

U(r^): the radial part of the Is orbital for the ith electron.

O

Yo(8i#^i): the angular part of Is orbital for the ith electron which is equal to^i

V(r^): the radial part of the 2p0 orbital for the ith electron.

O

Yi (® i^ i> : the angular part of the 2p0 orbital for ith electron which is equal to cos 0^.

so the total wavefunction for the Is orbital with a spin for the ith

JL O ]_

electron can be expressed as S = Y0 (0 <t> U (r^) a ~ tT U(ri)ai' and

the space part of the wavefunction is S(i) ~ U(r^). Similarly,

*o(i) = Yx (6 i#ai)v(ri)ai cos e ^ (r±) a± and P0(i) cos eiV(ri).

The inverse interelectron distance, — — , can be expressed as follows

j

(Eyring, Walter and Kimble, 1947) :

ID

L

k=o

+k

I

m=-k

4tt

2k+l k+1

<V

<t> )

m

We define the following integrals:

S(1)P0 (1) ) = JV;lF^ U(r1)V(r1)Y0 (61,*1)Y1(ei,*1)Sineid01d*1rJdr1

Pa n — 1 /Z Expan sio n

121

+k

= I E

4t r

J |U(r, )V (r, ) r. 2dr.

. . 2k+l ° „ k+1 1 ' 1 1 1

k=o m=-k r>

TT 2lt

7i 2tt -j ° in *

^ o o (6 9 ^ 1 ) Y]^ ( 9 9 $ 2. ) S 0 ^ 2f ^ 2^

The above reduces to:

| S (1) P0 (1) ) ='/|^yi(02^2){ r2 J o2r^U(r1)V(r1)dr1+r2^r U(r1)V(r1)dr1>

2 2

since the spherical harmonics form an orthonormal set, i.e.

jrjIYk*(0#<t,)Yk'(0^)Sin0d0d4) = ^k’^m'

(S (2) S (2) I S (1) P0 (1) ) = < S (2) S ( 2) \~~\ S(1)P0 (1)

12

/ - oo r oo

*3 J°U(r2){r2 S° riU(r1)V(r1)dr1 + r2 ' Ufr-^V (r^ dr^ r2dr2

r\2lT o

X;0J o Y-, (9 0,4> 0) Sin6 2d0 2d4> 2 = 0,

TT 2TT J J

because 0J0 (9 2, <P 2) Sin9 2d0 2d<f> 2 = 0 therefore the integral vanishes,

In general,

| SP0 ) ='/y- Y! ( d*<t> ){^2 1 ox3u (x)V (x)dx + r JU(x)V (x)dx) ,

r r

( S S | SPD ) = 0 and (P0P0|SP0) = 0 due to the orthonormal property of the spherical harmonics.

, 1 ^ . 2 TT 0 Q °°

However, (SP0|SP0) 3 * 1 0 <, Y1 (0 ,<J> ) (0 ,<|> ) Sin0 de d<p 1 CU (r )V (r)

x{

— 0 ; qX3U (x)V (x) dx + r 1 QU (x)V (x) dx r2dr.

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Transactions Illinois Academy of Science

CO _ w

or (SP0|SP0) = 1 0U (r)V (r) {— 9 J „xu (x)V (x) dx + rf U (x)V (x) dx}r2dr,

J r r

TT 2TT

o o

because [ £ (0 , <)> ) (6 , <t> ) Sine de d<)> = 1

Similarly, we define

| S (1)S (1) ) = ;v ^ U2(r1)Y^ (91,<t»1)Yo (01,4>1)Sin01d01d^1r2dr1

1 12

00 4tt

= E l

00 r <

TT 2 TT

, , 2k+l

k=o m=-k r

K k+1 U2(r1)r2dr1^0 '0 Y0 (0 ±. <j>1) Ye (9 r 4^) y£( 0 v 4^)

x Sineid61d^1Y^,C(e2<!)2) = ^ J^2 U2(r1)r2dr1 + J r U2 (r^ r1dr1

71 2 ^ 0 O " o

So, (SS | PQ P0 ) = (P0P0|SS) = J0J0 Y1(0,^)Y1(0,4>)Sin0ded4» J0v (r)

X[— J u2(x)x2dx + 1 v2 (x)xdxJ r2dr

= J ”2(r)[^ Jfu2(x)x2dx + JrU2 (x)xdx]r2dr,

_ 00

and (SSlSS) = J 0U2 (r ) [^! fu2 (x) x2dx + 5 U2 (x) xdxJ r2dr .

In general, (P0P0|SS) = J 0 v 0 . . r

1/Z Expansion of the Angular and Radial Hartree-Fock Equations

We use a unit of length equal to 1/7. A.U., a unit of energy 2

equal to Z A.U. and the notations of the preceding section. The Hamiltonian of this system can be written as

H = I h(i) + ^ z g (i, j )

i z ±<j

Pan — 1/Z Expansion

123

where h(i)

1 „ 2 2Vi “

1

r .

i

and

£ g(i»j) = + 7^- + ~~

i <j r12 r23 r 13

In order to obtain the total energy of this atomic state, we have to evaluate the diagonal matrix elements of the Hamiltonian of this system over the restricted Hartree-Fock wave funct ions . Let us first

3 a ^

consider the matrix elements of I h(i). Let be the permutation operator and ^ be the number of permutation.

Then, after multiplying by inverse permutations and their corresponding signs to eliminate the normalization factor (Eyring, Walter and Kimble, 1947) we have.

A(i)

£(i) s(i) £0(i)

i(2) S (2) £0(2)

£(3) 3(3) £0(3) = <£(1) |h(l) | S (1) > + <S(2) |h(2) | S ( 2) > + <£0 (3) |h (3) | £0 (3)>

<|S§f0|E h(i)|^J> Z (-1)^1^J^(1)S(2)^0 (3)

= 2<S| h| S> +<PG| h| P0> ;

where <S(i)|s(i)> = <S(i)|£(i)> = 0 and <§(i)|h(i)|s(i)> =

< S (i) | ft ( i) | £ (i)> = 0, because of spin orthogonality; < £0 (i) | i-5 (i)> =

< £ (i) | (i)> = 0 and < (i) | (i) | £ (i)> = < la (i) | h (i) | £0 (i)> = 0, because

of space orthogonality; <£0 (i) |^(i) | S (i)> = < S ( i) | $1 ( i) | £0 (i) > = 0, because of spin and space orthogonality;

124

Transactions Illinois Academy of Science

(i) |li(i) |§ (i) > = <S (i) |ft(i) I S (i) ><a (i) | a (i) > =

<P0|h|P0>. For matrix elements of 1/Z arguments to get the following results.

<|£s$0|£ Z g..|£s£0|> = i^(l)S(2)^0(3)g

'Z . .-in i<1 J

12

lh(i)

> =

j( we

can use similar

£(1)

S(l)

foU)

£< 2)

S(2)

f„(2)

dV

£(3)

S(3)

S(3)

123

£(i)

S(l)

fo(l)

|iS+(l)S(2)P0(3)g13

£(2)

S(2)

fo(2)

dV123

1 ■ y

£(3)

S(3)

J„(3)

| - »

S(D

S(l)

fo(l)

+ |;£(1)S(2)£0 (3)g23

S(2)

S(2)

fo(2)

^123

S(3)

S(3)

K (3)

The above equation can be simplified by multiplying the third orbital ($G) into the third row in the first term, the second orbital (S) into the second row in the second term and the first orbital (£) into the first row in the third term, as shown by the arrows. After using the orthonormality argument, we have the following results:

Pan — 1/Z Expansion

125

£s$0 || I g I £§P0 I > z k<j 30

J J(1)S (2)g12

+ J^(l)f0(3)g13

+ ;S(2)f0(3)g23

£(1)

S(l)

S(2)

S(2)

^ (1)

^o(D

S(3)

$o(3)

S(2)

(2)

S(3)

^o(3)

dV

12

dV

13

^23 }

= |(SS+SS) +|(ss|p0p0) -|(spo|p0s) +|(ss|p0p0)

= - (SS | SS) + 2 (SS | P0 P0 ) - (SPe | P0S) }

So, the total energy of (ls22p is

E(ls22p,22p) = 2(s|ti|s) + (P0 | h | P0 ) +|(SS|SS) +|(SS|P0P0) -|(SPo|P0S)

(1)

The orthonormality conditions are

;S2dV = 1, JP02dV = 1 and JSP0dV = 0,

Applying the variational principle to obtain the Hartree-Fock equa¬ tions, we use a star * to label the quantities being varied in Eq. (1). We have then

E = 2 (S | h | S ) + (Pe | n | P0 ) + ^(S S|S S) + |(S S |po P) - |(S P0 |po S)

(2)

126

Transactions Illinois Academy of Science

and we note that

6{E-2%s(S* |s)-xp(p0* |P„)-Xsp(S* |Pj-*ps(P„* Is)} = 0 is the same as 6{E-2Xs(S*|S)-Xp(P„*|P)-X*sp(S|P0*)-X*ps(Po|S*)} =0, since xsp

^p

is real. X = X where X's are Lagrange multipliers and (S|S), sp ps

(P0|P0) and (S | P0 ) are the constraints.

•fp

Carrying out the variation with respect to 6s , 6P0 , (and to 6s, aP0 separately) ; we have,

2(6S*|ft|s) + («P0*|ft|P.) +|(«S*S|S*S) + f(5S*S|P0*P„>

+ |(S*s|«P.*P0)- i(Ss*p0|p0*s)- |(S*PC,|6P0*S)

- | Xs(5S*|S)-Xp(«P0*|p<>)-Xsp({S*|P0)-Xps6 P0*|S) = 0

•fp *fp

where X = X Equating to zero the coefficients of 6s and 6P0 ,

sp ps

we obtain the Hartree-Fock equations:

fcs + \ S | SS) + \ S|P0P0) - ^2 Po|PoS) = ^sS + ^spPo _ (3)

kPo + | Polss) - | S | SP0 ) = XpPe + XpsS _ (4)

Since S and PG are orthonormal, we do not need to introduce the Lagrangian multipliers Xgp and Xpg; therefore, we set Xgp = Xpg = 0, and Eqs. (3) and (4) become

hS + | S | SS) + \ S | P0P0 ) - -|ZP0|P0S) = XsS _ (5)

1po + I p0|ss) - \ s|sp0) = xpPo _ (6)

These equations depend on the parameter Z'1 as well as the X’s. S and P0 can be obtained as functions of that parameter by solving these equations. The present procedure also can be used to derive regular Hartree-Fock (restricted or unrestricted) equations. The derivation of the regular Hartree-Fock equations can be considered as a special case of the present method. All procedures are the same, except that we omit the 1/Z factor in the equations.

Pan — 1/Z Expansion

Acknowledgment

127

The author would like to acknowledge helpful discussions with Dr. A. Golebiewski of the Department of Theoretical Chemistry, Jagiellonian University, Poland and Pro¬ fessor Y. N. Chiu of the Catholic University of America. He also wishes to thank Dr. Dennis Sardella of Boston College for reading the manuscript.

Literature Cited

Cohen, M. and A. Dalgarno. 1961. Stationary Properties of the Hartree- Fock Approximation. Proc. Phys. Soc. (London). 77: 748-750.

- . 1963a. An Expansion Meth¬ od for Calculating Atomic Properties III. The JS and 2P° States of the Lithium Sequence. Proc. Roy. Soc. (London) A275 : 492-503.

- . 1963b. On the Orthogonal¬ ity Problem for Excited States. Revs. Mod. Phys. 35: 506-508.

- . 1964. An Expansion Meth¬ od for Calculating Atomic Properties IV. Transition Probabilities. Proc. Roy. Soc. (London) A280: 258-270. - . 1966a. An Expansion Meth¬ od of Calculating Atomic Properties.

VIII. Transitions in the Hartree-Fock Approximation. Proc. Roy. Soc. (Lon¬ don). A293 : 359-364.

- . 1966b. An Expansion Meth¬ od for Calculating Atomic Properties.

IX. The Dipole Polarizabilities of the

Lithium Sequence. Proc. Roy. Soc.

(London) A293: 365-377.

- . 1967. An Expansion Meth¬ od for Calculating Atomic Properties.

X. Is2 ^-lsnpT Transions of the Helium Sequence. Proc. Roy. Soc. (London). A301 : 253-260.

Coulson, C. A. and A. Hibbert. 1967. Z~V2 Expansion of the Unrestricted Hartree-Fock Equations for the (Is) (Is') 'S States of Helium and Helium- Like Ions. Proc. Phys. Soc. (London). 91: 33-43.

Dalgarno, A. 1960. The Correlation Energies of the Helium Sequence. Proc. Phys. Soc. (London). 75: 439-440: - . 1962. Atomic Polarizabili¬ ties and Shielding Factors. Adv. in Phys. 11: 281-315.

Hirschfelder, J. O., W. B. Brown and S. T. Epstein. 1964. Recent Develop¬ ments in Perturbation Theory. Adv. Quantum Chem. 1 : 255-374.

Eyring, H. J. Walter and G. E. Kimball. 1947. Quantum Chemistry. John Wiley & Sons, Inc., N.Y. XXIV + 432

pp.

Linderberg, J. 1961. Perturbation Treat¬ ment of Hartree-Fock Equations. Phys. Rev. 121: 816-819.

Schwartz, C. 1962. Importance of An¬ gular Correlations Between Atomic Elec¬ trons. Phys. Rev. 126: 1015-1019. Sharma, C. S. and C. A. Coulson. 1962. Hartree-Fock and Correlations Energies for ls2s 3S and *S States of Helium-Like Ions. Proc. Phys. Soc. (London). 80: 81-96.

- . 1962a. A Perturbation

Treatment of the Unrestricted Hartree- Fock Equations for the Ground State of Lithium-Like Ions. Proc. Phys. Soc. (London) 80: 839-848.

- . 1968. On Hartree-Fock In-

tergro-Differential Equations for Atoms. Proc. Roy. Soc. (London). 304. 513-530. Stewart, A. L. 1964. On the Z"1/2 Ex¬ pansion of Unrestricted Hartree-Fock Wave Functions. Proc. Phys. Soc. (Lon¬ don). 83: 1033-1037.

Manuscript received August 10, 1969

DEATH OF CELLS IN PITH TISSUE OF SOYBEAN

SEEDLINGS

KUO-CHUN LIU AND A. J. PAPPELIS

Department of Botany, Southern Illinois University, Carbondale , Illinois 62901

Abstract. — Parenchyma cell death in the hypocotyl pith tissue of soybean seed¬ lings was discovered in each of the 24 varieties studied representing seven ma¬ turity classes. Dead cells generally were observed on the fifth day after planting; and chlorenchyma tissue formed between the dead pith parenchyma cells and xylem in the above-ground stem. Dead paren¬ chyma cells were observed in the below¬ ground cortex and in the pith of the elon¬ gating internode above the cotyledons. No dead cells were observed in the cotyledonary node during the 20-day study period.

The death of cells in pith parenchyma of the hypocotyl and the first and second inter¬ nodes of seedlings of 18 soybean varieties was studied in relation to two or three planting depths (sub-surface, 2.5 cm, and 5.0 cm). Differences were noted in the rate of cell death in hypocotyls but not in epicotyl internodes as a result of planting depth differences. In the hypocotyls, most pith parenchyma cells died during the first week of growth, and the greater depth of planting resulted in the fastest rate of cell death in each variety. Cell death in pith parenchyma of internodes occurred as the internodes elongated.

Patterns of parenchyma cell death have been reported for normal and injured sorghum (Katsanos and Pap¬ pelis, 1969), sugarcane (Pappelis and Katsanos, 1965), and corn (Pappelis and Katsanos, 1969). It was considered desirable to seek simi¬ lar parenchyma death in a plant more suited for study in growth chambers. This paper reports the discovery of parenchyma cell death in soybean seedlings and describes variations in cell death patterns as¬ sociated with variations in planting depth.

Materials and Methods

Twenty-four soybean varieties

(Class 00, Acme, Flambeau, Portage ; Class 0, Grant, Merit, Norcheif ; Class I, A-100, Chippewa, Chippewa 64, Ontario ; Class II, Hawkeye, Lindarin ; Class III, Adams, Ford, Harosoy, Shelby, Wayne; Class IV, Clark, Kent, Midwest, P.I. 84.946-2, Clark 63; Class V, Dorman, Hill) were grown under greenhouse con¬ ditions at various times from Decem¬ ber 29, 1964, to March 31, 1965. Seeds of each variety were planted in separate wood flats 2.5 cm apart at a depth of 5 cm in a soil mixture of 50% sand and 50% peat moss. Five plants from each variety were selected for study each day begin¬ ning at the second day after plant¬ ing and continued until the first in¬ ternodes had elongated. The maxi¬ mum study period was 20 days.

Twelve soybean varieties were planted (April 29, 1965) in sepa¬ rate wood flats, 2.5 cm apart at depths of 2.5 or 5.0 cm. Watering was accomplished with a mist spray¬ er to reduce soil packing or washing. Five seedlings of each variety were selected from each planting depth on the seventh, tenth, and twentieth days after planting. Also, seedlings of six varieties were studied after planting (May 20, 1965) at three depths : 5.0 cm, 2.5 cm, and just below the soil surface. For the lat¬ ter, seeds were placed on the soil surface and covered with a thin layer of soil to permit germination below the soil. Watering and sampling procedures were as those described

[128]

Liu and Pappelis — Cell Death in Soybean Stems

129

above but with two samples studied seven and fourteen days after plant¬ ing.

Dead cells in liypocotyl and epi- cotyl pith tissue were discovered in cross section and longitudinal section using the plasmolyzing neutral red stain solution described by Tribe (1955).

The length of white liypocotyl tis¬ sue of the below-ground part was used as the final estimate of plant¬ ing depth. The lengths of hypo- cotyls and internodes were meas¬ ured before the stem was cut longi¬ tudinally. Lengths of pith com¬ posed of dead cells were measured and the per cent of total length with dead cells calculated.

Results

Dead and living pith cells were easily distinguished microscopically

in cross and longitudinal sections of the hypocotyls using the plas- molyzing-neutral red stain. In stained sections, living cells con¬ tained dark-red, plasmolyzed proto¬ plasts, while dead cells were light in color and contained no plasmol¬ yzed protoplasts. In every case, dead cells contained a gas bubble. No remains of the protoplast or cell organelles were observed. When masses of dead cells occurred in the pith tissue, macroscopically the pith appeared white in color and was spongy. Tissue composed of living cells was well hydrated and appeared light green in color in the above¬ ground part, and cream in color in the below-ground part.

In all varieties dead cells were first observed in the central area of the below-ground liypocotyl pith five days after planting (Figure 1). Sev-

Figure 1. Dead pith parenchyma cells occur in hypocotyls five days after plant¬ ing. From left to right, appearance of seedlings and areas of dead cells at 4, 5, 6, 8, and 10 days after planting; dotted areas representing areas of dead parenchyma cells in the pith.

130

Transactions Illinois Academy of Science

eii to eight days after planting, al¬ most all of the cells in pith tissue of the below-ground hypocotyl were dead. Ten days after planting dead cells were observed in the central region of the pith of the above¬ ground hypocotyl and a hollow area formed in the below-ground hypo¬ cotyl pith. This was associated with an increase in below-ground diame¬ ter of the hypocotyl ; a result of cortical expansion. At this stage, dead cells and hollow areas were also observed in the cortex. In the above¬ ground hypocotyl, pith cells adja¬ cent to the xylem developed into a layer of chlorenchyma, four to five cells in thickness.

In the first internodes of all vari¬

eties studied, dead cells appeared in the pith as the internodes elongated. A chlorenchyma layer developed in the intemode pith adjacent to the xylem tissue. No dead cells ap¬ peared in the cotyledonary node or in nodes above this point.

In the two experiments designed to determine the effects of planting depth on cell death in stem tissue, little differences were noted in the cell death patterns in the epicotyl intemodes. For this reason, only the results from two varieties, one from each experimental group, will be presented (Table 1). In the hy- pocotyls, pith cells died in different patterns due to planting depth. The data for 12 varieties, planted at 2.5

Table 1. — Cell death percentages in the hypocotyl, first internode, and second internode pith tissue at different stages of seedling development from seeds of Harosoy planted at two depths and Kent planted at three depths. (% DC = per cent of the area of pith tissue with dead cells.)

Seedling Part

Depth

Hypocotyl

First Internode

Second Internode

of

Days

Planting

(cm)

After

Planting

Length

% DC

Length

% DC

Length

% DC

Harosoy (Planted April 29)

2.5

7

6.6

45

1.7

0

10

7.0

85

5.3

5

20

7.7

87

8.5

96

4.0

0

5.0

7

8.1

56

1.9

0

10

8.2

92

6.2

8

20

8.0

96

9.2

96

4.4

27

Kent (Planted May 20)

Sub-surface

7

4.5

0

2.5

0

14

4.4

52

11.3

91

7.2

54

2.5

7

6.0

50

4.8

0

14

5.9

76

12.4

84

7.8

57

5.0

7

8.2

88

5.0

4

14

8.7

95

11.7

99

7.5

49

Liu and Pappelis — Cell Death in Soyhea?i Stems

131

and 5.0 cm, and sampled 7, 10, and 20 days after planting- are given in Table 2. The data for 6 varieties, planted at sub-surface, 2.5 cm, and 5.0 cm, and sampled 7 and 14 days after planting are given in Table 3.

In the study of cell death in 12 varieties at two planting depths, seven days after planting, the lengths of hypocotyls of seedlings planted at 2.5 cm depths ranged from 6 to 9 cm. During the next 13 days, additional growth was generally less than 1 cm ; Acme being most extreme with an additional 1.9 cm of growth. Hypocotyls of seedlings at 5.0 cm depth ranged in length from 7 to 9

cm 7 days after planting and in¬ creased about 1 cm in length during the next 13 days, Hawkeye being- most extreme with an additional 2.5 cm of growth. The length of hypo- cotyl with dead pith cells on the sev¬ enth day after planting ranged from 41 to 65% for seeds planted at 2.5 cm depth and 53 to 88% for those plant¬ ed at 5.0 cm depth. In all but one case, the seedlings from the deepest planting depth had the highest per¬ centage of dead cells in each variety. By the twentieth day after planting, the amount of dead cells in the pith ranged from 88 to 100% in the plants from both planting depths

Table 2. — Per cent of the area of pith tissue of hypocotyls composed of dead cells in seedlings of 12 varieties of soybeans planted at two depths and sampled three times after planting (April 29).

Variety

A-100 .

Acme .

Chippewa 64 Chippewa. . .

Clark .

Dorman .

Flambeau . . .

Ford .

Grant .

Harosoy . Hawkeye. . . Hill .

Depth

of

Planting

(cm)

Per cent of hypocotyl with dead cells; days after planting

7

10

20

2.5

53

96

96

5.0

66

99

96

2.5

42

95

96

5.0

53

96

98

2.5

64

61

97

5.0

88

97

97

2.5

44

96

96

5.0

68

98

98

2.5

46

84

95

5.0

66

75

96

2.5

57

94

96

5.0

64

94

95

2.5

41

97

100

5.0

66

97

96

2.5

57

96

97

5.0

64

96

97

2.5

61

71

90

5.0

57

95

95

2.5

45

85

87

5.0

55

95

96

2.5

50

40

97

5.0

62

80

98

2.5

58

75

91

5.0

66

68

88

132

Transactions Illinois Academy of Science

with no apparent difference due to the depth of planting’. Generally, most of the pith parenchyma cells had died before the tenth day after planting.

The above-ground growth in the 12 varieties showed some variations but within each variety, the growth of the first and second internocles and the per cent of pith tissue with dead cells were similar in the seed¬ lings from both the 2.5 and 5.0 cm planting depths. The average growth of the first internodes seven days after planting ranged from none vis¬ ible to 1 cm, 4 to 10 cm on the tenth day, and 6 to 14 cm on the twentieth day after planting. The second inter¬ nodes began to elongate between the tenth and twentieth days after planting, ranging in size from about 3 to 7 cm on the twentieth day. Cell death in the first internodes oc¬ curred after the seventh day. On the tenth day, the amounts of dead cells in the pith ranged from 0 to 43% in seedlings from the 2.5 cm depth and 8 to 55% in those from the 5.0 cm depth. On the twentieth day after planting, the amounts of pith with dead cells in the first internodes ranged from 92 to 98% in seedlings from the 2.5 cm depth (except Grant, 45%) and 95 to 98% in those from the 5.0 cm depth. The amounts of dead cells in the pith tissue of the second internodes ranged from 0 to 82% in seedlings from the 2.5 cm depth and 25 to 82% in those from the 5.0 cm depth. Generally, death of pith cells began in the lower part of the pith tissue of the internodes after they had grown a few centimeters in length. The slight differences noted in lengths and percentages of pith

length with dead cells for Harosoy grown at two planting depths (Ta¬ ble 1) are typical for the 12 varieties studied.

The effects of sub-surface, 2.5, and 5.0 cm planting depths on cell death in stem tissue were most noted for hypocotyl pith. The results with Kent (Table 1) were typical both for hypocotyl and epicotyl pith cell death observed in all six varieties.

For all six varieties, hypocotyl lengths from the sub-soil plantings ranged from 4 to 6 cm on the sev¬ enth day and had little additional growth. The percentages of dead cells ranged from 0 to 84% on the seventh day and from 5 to 92% on the fourteenth day (Table 3). When these six varieties were planted at a depth of 2.5 cm, hypocotyls at 7 days after planting ranged in length from 5 to 7 cm with little or no addi¬ tional growth during the next week. The amount of dead pith parenchy¬ ma ranged from 38 to 92% 7 days after planting and 14 to 93% 14 days after planting. The hypocotyls of the seedlings of the six varieties planted at 5.0 cm depths ranged in length from 7 to 12 cm 7 days after planting with little additional growth occurring during the next week. The ranges of cell death per¬ centages for the hypocotyl pith tis¬ sue were 88 to 93% on the seventh day and 95 to 97 % on the fourteenth day after planting. For all six va¬ rieties planted at sub-soil depth, the first internode lengths ranged from 3 to 11 cm 7 days after planting with 0 to 43% of the pith parenchy¬ ma cells dead, and 9 to 13 cm 14 days after planting with 80 to 100% of the cells dead. The second inter¬ nodes began to elongate during the

Liu and Pappclis — Cell Death in Soybean Stems

133

Table 3. — Per cent of the area of pith tissue of hypocotyls composed of dead cells in seedlings of six varieties of soybeans planted at three depths and sampled two times after planting (May 20).

Variety

Depth

of

Planting

(cm)

Per cent ol with dead after p

7

hypocotyl cells; days [anting

14

Kent .

sub-surface

0

52

2.5

50

76

5.0

88

95

Lindarin .

sub-surface

37

45

2.5

66

82

5.0

93

96

Merit .

sub-surface

17

5

2.5

38

14

5.0

88

96

Norcheif .

sub-surface

32

92

2.5

62

93

5.0

96

95

Portage .

sub-surface

58

58

2.5

66

46

5.0

97

97

Wayne .

sub-surface

84

58

2.5

92

90

5.0

89

97

second week and ranged from 5 to 8 cm in length 14 days after plant¬ ing with 39 to 60% of the pith parenchyma cells dead. For the six varieties planted at a depth of 2.5 cm, first internodes lengths ranged from 5 to 13 cm 7 days after plant¬ ing and 7 to 11 cm one week later. The dead cell percentages ranged from 0 to 38% and 84 to 99% on the seventh and fourteenth days, re¬ spectively. The second internodes began to elongate after the first sam¬ pling and ranged in length from 6 to 9 cm on the fourteenth day with cell death percentages ranging from 39 to 79%. For the six varieties planted at a depth of 5.0 cm, the first internode lengths for the six varieties ranged from 4 to 8 cm on

the seventh day with 3 to 29% of the length containing dead cells and 8 to 17 cm on the fourteenth day with 68 to 99% of the lengths con¬ taining dead pith cells. The second internodes began to elongate during the second week after planting and, on the fourteenth day, the lengths ranged from 7 to 9 cm with cell death averages ranging from 49 to 74%.

No death of pith cells was ob¬ served in nodal areas in any of the experimental groups.

Discussion

The death of parenchyma cells in the pith tissue of soybean occurred in all varieties studied, regardless of maturity class. Dead cells first

134

Transactions Illinois Academy of Science

appeared in the central rows of pith cells in the below-ground part of the hypocotyl, usually within 5 days after planting. In all three plant¬ ing groups, dead cells were observed in the pith of the above-ground and below-ground parts of the hypocotyl within two weeks. Hollow areas sometimes formed in the pith of the hypocotyl, especially in areas where increased cortical diameter had oc¬ curred. Whether expansion caused cortical cells to die is not known.

Death of pith parenchyma in the first and second internodes appeared after internode elongation began. Death of cells was first observed in the lower part of the inter node.

In both planting depth experi¬ ments, the patterns for cell death were like those observed for all va¬ rieties at one planting depth, death of cells first occurring in the lower part of the internode or hypocotyl (Fig. 2). The death patterns were similar in all varieties studied but the rate of cell death differed be¬ tween varieties.

In the study of the six varieties at 3 planting depths, it was very clear that the differences in growth and cell death in the first and sec¬ ond internode of any variety was very small. When compared as a group, some differences were noted in growth or cell death trends be¬ tween varieties, but these too were small. The obvious differences in each variety were the percentages of cell death in pith cells of the hypocotyls of seedlings planted at the three depths, the greatest amount of cell death occurring at the deep¬ est planting depth and the least

amount occurring when the seeds were planted just below the soil sur¬ face. Varietal differences were not¬ ed (Table 3), with cell death per¬ centages in Norcheif showing little difference due to planting depth while great differences occurred in Merit. Since cell death patterns in parenchyma tissue in stalks of corn, sorghum, and sugarcane is correlated with the areas susceptible to stalk rot in those crop plants (Ivatsanos and Pappelis, 1969 ; Pappelis and Ivatsanos, 1965 and 1969), it may be that the patterns of cell death dis¬ covered in the stem of soybeans will provide a new approach to the study of the nature of resistance to spread of Ceplialosporium gregatum Ailing- ton and Chamberlain, the causal or¬ ganism for the brown stem rot dis¬ ease in soybean.

Acknowledgment

We wish to thank Dick Bernard, U.S. Regional Soybean Laboratory, Urbana, Illi¬ nois, for providing the seed used in this study.

Literature Cited

Katsanos, R. A. and A. J. Pappelis. 1969. Relationship of living and dead cells to spread of Colletotricum graminicola in sorghum stalk tissue. Phytopathology 59:132-134.

Pappelis, A. J. and R. A. Katsanos. 1965. Spread of Physalospora tucumanensis in stalk tissue of sugarcane. Phytopathology 55:807-808.

- . 1969. Ear removal and

cell death rate in corn stalk tissue. Phytopathology 59:129-131.

Tribe, H. T. 1955. Studies on the physi¬ ology of parasitism. XIX. On the kill¬ ing of plant cells by enzymes from Botrytis cinerea and Bacterium aroideae. Ann. Bot. 19:351-369.

Manuscript received November 30, 1970

Liu and Pappclis — -Cell Death in Soybean Stems

135

Figure 2. Effect of planting depth on the death of cells in pith tissue of soybean seedlings. From left to right, seedlings from sub-surface, 2.5 cm, and 5.0 cm depths, respectively. Dotted areas in the stem represent the areas (averages of six varieties) where dead cells occurred one week after planting (May 20; greenhouse conditions; equal parts sand and peat).

MICROBODIES OF SOYBEAN COTYLEDON MESOPHYLL

KUO-CHUN LIU, A. J. PAPPELIS, AND H. M. KAPLAN

Department of Botany, and Department of Physiology, Southern Illinois University, Carhondale, Illinois 62901

Abstract. — Microbodies were found in the upper mesophyll cells of soybean [ Glycine max (L.) Merr., var. Wayne] cotyledon from the germinating stage (24 hr after planting) to cotyledon abscission (12 hr day, 12 hr night, 25°C, and 700 ft-c). These organelles were associated with endoplasmic reticulum, mitochondria, and chloroplasts. The shape of these or¬ ganelles changed from oval, circular, or elongate in the earlier stage of seedling development to circular forms when cotyle¬ dons became yellow. The microbodies in senescent cells often lacked a continuous bounding membrane.

The occurrence, structure and the enzymatic function of plant micro¬ bodies is a subject of increasing in¬ terest to many investigators. Some of the characteristics of plant micro¬ bodies can be summarized as follows : thev are bounded by a single mem- brane ; they have a diameter from 0.5 to 1.5 [). ; a dense granular stroma ; and they are often associated with endoplasmic reticulum. Some are re¬ ported to contain crystals. These characteristics apply equally to two types of cell particulates isolated from homogenates. One of these, ob¬ tained from leaves, is involved in photo-respiration and is referred to as a peroxisome. The second, ob¬ tained from endosperm, is involved in the formation of succinate from fatty acids and is referred to as a glvoxysome. In cotyledons, although glvoxysomes appear early and are

replaced by peroxisomes, the cyto- logical events associated with these changes are unclear. Much of the literature describing plant micro¬ bodies and their associated enzvmes has recently been reviewed (Beevers, 1969 ; Breidenbach, 1969 ; Gruber, et al., 1970; Tolbert and Yamazaki, 1969; and Vigil, 1970).

No description of soybean micro¬ bodies has yet been published. In our recent study of cellular senes¬ cence in soybean cotyledons, we found organelles similar to published electronmicrographs and descrip¬ tions of glyoxysomes and peroxi¬ somes. This paper presents some of our observations.

Materials and Methods

Soybean [Glycine max (L.) Merr., var. Wayne] seeds were planted at a depth of 5 cm in sand and peat mixture (1:1 by volume) and grown in a growth chamber (25° C both for 12 hr of 700-ft-c of light and 12 hr of dark period). Sampling was at 24 hr intervals after planting for six days (at which time the hypo- cotyls were about 5 cm above the soil surface). On the sixth day, seed¬ lings were standardized for uniform height and for uninjured cotyle¬ dons. All other seedlings were re-

[136]

Liu ct al. — Soybean Cotyledon Microbodies

137

moved. After the sixth day, sam¬ pling was obtained at three-day in¬ tervals until cotyledon abscission.

For each sample, tissue blocks of 1x1x4 mm were cut from the upper central area of the cotyledon. The blocks were fixed with 3% glu- taraldehyde in 0.066 M phosphate buffer at pH 7.4 for 4 hours. After rinsing three times with the same buffer, the tissues were post-fixed in a 1 :1 mixture of 2% osmium tetrox- ide and the above buffer for two hours. Both fixations were at room temperature. The tissues were de¬ hydrated through an ethanol series, treated with propylene oxide and embedded in Epon 812 (Luft, 1961).

Sections were obtained with a dia¬ mond knife on a Reichert Om-U2 ultramicrotome, mounted on 200 mesh copper grids, stained in uranyl acetate (Watson, 1958), and coun¬ ter-stained with lead citrate (Rey¬ nolds, 1963), and examined with a Hitachi HU-11A electron microscope at 50 KV.

Results

We did not find microbodies in cotyledons sampled 24 hrs after planting. Single membrane bound¬ ed electron dense, granulate organ¬ elles (Figs. 1-3) were observed in samples obtained at 48 hr after planting and in all other subsequent samples. Endoplasmic reticulum was associated with or in the vicinity of those organelles. The diameters of the dense organelles ranged from 0.5 to 1.3 /x, which is in the range of plant microbodies. We conclud¬ ed that these were microbodies.

The shapes of the microbodies were spherical, elongated, irregular, or

dumbbell (Figs. 2-6). Elongated microbodies had diameters of 0.3 y at the narrowest region to 1.3 y at the widest region. The stroma ap¬ peared as electron dense granules with transparent areas scattered within it. A single membrane was clearly identified as surrounding the microbodies of the earlier samples (Figs. 1-4). The bounding mem¬ brane of the microbodies from the sample of yellow cotyledon (21 days after planting) could not be seen clearly, but the endoplasmic reticu¬ lum associated with the organelles persisted (Figs. 5-6). The diameters of the microbodies in senescent cells were about 0.5 to 1.0 y.

In addition to the close associa¬ tion between the microbodies and endoplasmic reticulum, in most of the cells the microbodies and chloro- plasts were appressed (Figs. 3-4), changing the shape of the microbody at the place where the close spatial association occurred. Similar close associations between microbodies and mitochondria, and between mito¬ chondria and chloroplasts were also observed.

Discussion

In fatty cotyledons, stored lipids are converted to carbohydrates in the early post-germinative stages. The microbodies observed in the soy¬ bean cotyledons at this stage were assumed to be glyoxysomes. As the cotyledons expand and emerge from the soil, they become photosynthetic organs. The microbodies observed in soybean cotyledons associated with the chloroplasts at this stage of growth are assumed to be peroxi¬ somes. A time must exist when both

138 Transactions Illinois Academy of Science

PLATE I

Figure 1. Two microbodies (Mb) among lipid bodies (L) in a cotyledon from three day old seedling; Cw, cell wall, x26,000.

Figure 2. Microbody (Mb) with endoplasmic reticulum (ER) in a cotyledon from four day old seedling. Mitochondrion (M), chloroplast (C), and starch strain (S) also can be identified, x26,000.

Figure 3. Microbody (Mb) associated with endoplasmic reticulum (ER) and developing chloroplast (C) in a cotyledon from five day old seedling, x33,000.

Figure 4. Microbody (Mb) with endoplasmic reticulum (ER), and chloroplast (C) in a cotyledon from a ten day old seedling, x52,000.

Liu ct al. — Soybean Cotyledon

Microbodies

139

PLATE II

Figure 5. Microbodies (Mb) associate with endoplasmic reticulum (ER) from yellow cotyledon of 21 day old seedling. Degenerated mitochondrion (M) and degen¬ erated chloroplasts with electron dense globules (G) also can be seen, x35,000.

Figure 6. As Figure 5, x44,000.

140

Transactions Illinois Academy of Science

types of microbodies exist within the same cell of a cotyledon. Gruber et al. (1970) showed that micro¬ bodies are present at three distinct stages of cotyledon development of sunflower, cucumber, and tomato. The microbodies progress from cata¬ lase-containing particles located among lipid and protein bodies, to glyoxysomes closely associated with lipid bodies, to peroxisomes frequent¬ ly appressed to chloroplasts. Al¬ though a transitional period oc¬ curred involving decline of glyoxy¬ somes and a rapid rise of peroxi¬ somes, the origin of the particles and their mode of destruction were not described. They did not determine whether any of the particle types were derived from preexisting mi¬ crobodies or whether each arise as a separate population.

Vigil (1970) demonstrated that castor bean cotyledon microbodies showed membrane continuity with rough endoplasmic reticulum, sug¬ gesting a mode of formation similar to that in animal cells. As cotyle¬ don development progresses, some microbodies disappear in toto by se¬ questration into autophagic vacuoles. The loss of enzymes did not appear to occur prior to digestion of the sequestered microbodies.

The rapid loss of lipids during germination can be seen in Figs. 1-4, the latter being sampled five days after planting. Starch was observed in developing proplastids at the stage of germination when seedlings were still underground. The glu¬ cose for this starch synthesis was believed to be derived from the con¬ version of lipids to liexose in a proc¬ ess involving glyoxysomes. The close association of microbodies and chlo¬

roplasts at a later stage is inter¬ preted to imply that peroxisomal photorespiration occurs in soybean cotyledons.

The change in shape of plant mi¬ crobodies from oval, elongate, and irregular in young tissue to almost perfectly circular (Figs. 5-6) in old tissue, and the inability to visu¬ alize a distinct bounding membrane, are the only noticeable changes dur¬ ing senescence. The chloroplast changes during aging (osmiophilic bodies, disruption and loss of grana stacks, and the appearance of cir¬ cular fragments of lamellae) are similar to those reported by Barr and Arntzen (1969).

Acknowledgment

This research was supported by funds to Physiology Research and to Interdisci¬ plinary Research in Senescence, a Special Project and a Cooperative Research Proj¬ ect, respectively, of Southern Illinois Uni¬ versity. We wish to thank John A. Rich¬ ardson, Scientific Photographer and Illus¬ trator, Research and Projects, Graduate School, for his assistance in preparing the photographs for publication.

Literature Cited

Barr, R. and C. J. Arntzen. 1969. The occurrence of 5-tocoherylquinone in high¬ er plants and its relation to senescence. Plant Physiol., 44:591-598.

Beevers, H. 1969. Glyoxysomes of castor bean endosperm and their relation to glucogenesis. Annals New York Acad. Sci., 168:313-324.

Breidenbach, R. W. 1969. Characteriza¬ tion of some glyoxysomal proteins. An¬ nals New York Acad. Sci., 168:342-347. Gruber, P. J., R. N. Trelease, W. M. Becker, and E. H. Newcomb. 1970. A correlative ultrastructural and enzy¬ matic study of cotyledonary microbodies following germination of fat-storing seeds. Planta (Berk), 93:269-288.

Luft, J. H. 1961. Improvements in epoxy resin embedding methods. J. Bio- phys. Biochem. Cytol., 9:409-414.

Liu et al. — Soybean Cotyledon Microbodies

141

Reynolds, E. S. 1963. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell Biol., 17:208-212.

Tolbert, N. E. and R. K. Yamazaki. 1969. Leaf peroxisomes and their rela¬ tion to photorespiration and photosyn¬ thesis. Annals New York Acad. Sci., 168:325-341.

Vigil, E. L. 1970. Cytochemical and de¬

velopmental changes in microbodies (glyoxysomes) and related organelles of castor bean endosperm. J. Cell Biol., 46:435-454.

Watson, M. L. 1958. Staining of tissue sections for electron microscopy with heavy metals. J. Biophys. Biochem. Cytol., 4:475-478.

Manuscript received November 30, 1970

THIN FILMS OF INTERPHASE CHROMATIN PREPARED FOR ELECTRON MICROSCOPY BY OSMOTIC SHOCK

TECHNIQUE

FATHI ABDEL-HAMEED

Department of Biological Sciences,

Northern Illinois University, DeKalb, Illinois 60115 USA

Abstract. - — Interphase chromatin of chicken erythrocytes does not spread in a langmuir trough. By a new technique of osmotic disruption it spreads into a mass of knobby 70-625 A wide fibers which ag¬ gregate to form dense chromocenters. The new method is simpler and faster than the conventional one. Enzymatic digestions in¬ dicate that DNA is necessary for the linear continuity of nucleoprotein fibers and his¬ tone seems to be necessary for fiber aggre¬ gation and the condensation of chromatin.

With the advent of the electron microscope, it was anticipated that the internal organization of heredi¬ tary material in higher organisms could be resolved. But except for the synaptinemal complexes in paired meiotic chromosomes (Moses, 1960), examination of thin sections has offered very little information. Thus, the recent discovery by Gall (1963) of a method for spreading nuclear content into monolayer films in a “Langmuir trough” represents a technical breakthrough. Gall’s technique has been used successfully by several investigators to study the fine structure of interphase chroma¬ tin, and mitotic and meiotic chromo¬ somes in quite a few different or¬ ganisms (Bis and Chandler, 1963; Gall, 1966; Wolfe, 1965a, Wolfe, 1965b; Wolfe and John, 1965; Du- Praw, 1965). However, nuclei of certain cell types do not spread readily in a Langmuir trough. For example, chicken nucleated erythro¬

cytes and human sperm heads spread very little or not at all while those of amphibian erythrocytes and grass¬ hopper sperms spread well (Fig. la, b). Apparently, the former pos¬ sess a rather rigid membrane sys¬ tem. Removing the membranes of human or bull sperm heads by alka¬ line thioglycolate exposes their nu¬ clei and gives adequate chromatin spreading (Lung, 1968).

Chicken erythrocyte nuclei can be spread by osmotic shock (Fig. lb, c). This method is a modification of the protein monolayer technique devised recently by Freifelder and Kleinschmidt (1965) to spread iso¬ lated viral DNA.

Materials and Methods

One ml of rooster’s venous blood was collected in a heparinized sy¬ ringe and washed 3 times in 0.01 M Tris buffer containing 0.003 M MgCl2, pH 7.5. Red blood cells were sedimented at 200 g for 5 min. They were then resuspended in hypotonic solution or distilled water and cen¬ trifuged at 2000 g for 10 min. This step was repeated 2 or 3 times until an upper layer of intact free nuclei was obtained.

The free nuclei were suspended to a final concentration of about 105/ml in a hyper phase solution of

[142]

Abclel-Hameed — Interphase Chromatin

143

1.0 M ammonium acetate, 0.5% neu- livering* such a small volume was tralized formaldehyde and 0.01% obtained by using a 10 lambda Ep- cytochrome c (Nutritional Biochem. pendorf micropipette (Brinkmann Corp.). A volume of 0.01 ml of well Instruments).

mixed hyperphase was poured down The surface film was picked up an inclined wet glass slide into a by touching it with a Formvar-car- petri dish containing chilled hypo- bon coated copper grid. Immediate- phase solution of 0.3 M ammonium ly after picking up the surface film, acetate and 0.5% neutralized for- uranyl staining was achieved by maldehyde. Excellent results in de- dipping the grids in freshly pre-

Figure 1. (a) A single erythrocyte nucleus of the Mexican salamander Amby-

stoma mexicanum spread in a Langmuir trough into essentially a monolayer preparation. It shows the dense fibrillar components of interphase chromatin in these cells. X 2,090; (b) The erythrocyte nucleus of a chicken remained intact after being spread in a “Langmuir trough”. X 2,010; (C)A single erythrocyte nucleus of chicken spread by osmotic shock technique into a mass of knobby chromatin fibers. X 10,400.

144

Transactions Illinois Academy of Science

pared Wetmur ’s ethanol solution of the stain (Wetmur et ah, 1966) for 30 see. and then dehydrated for 15 sec. in chilled isopentane followed by air drying1.

In DNase, RNase or trypsin treat¬ ments, digestion was carried on prior to staining. Trypsin (Sigma, 2X crystallized) and RNase (Nutrition¬ al Biocliem. Corp. Crystalline) solu¬ tions were prepared as 1.0 mg/ml and 0.1 mg/ml of glass distilled wa¬ ter respectively. But DNase (Wor¬ thington deoxyribonuclease I, elec- trophor. purified) was prepared as 0.1 mg/ml of .003 M MgCl2 and .01 M Tris buffer, pH 7.5. Stock solu¬ tions of the hyperphase, hypophase, Wetmur ’s stain, and Tris buffer were all millipore filtered at least once. The grids were examined in a JEM- T7 at 60 IvV and an EMU-3 RCA at 50 IvV.

Results

Inter phase chromatin of chicken erythrocytes spreads into an inter¬ connected mass of 70-625 A wide nueleoprotein fibers. Except for the 70 A fibers which seem to be the basic structural unit of chicken chromosomes, two parallel or rela- tionally coiled sub-units were often observed within the wider fibers. At first glance the 625 A fibers seemed to consist of two 300 A units, the 300 A fiber of two 150 A units and the latter of two 70 A units. How¬ ever, fibers show definite polarity in their diameter tapering off from their base outward. They also ex¬ hibit a random pattern of branch¬ ing into, or association between, fi¬ bers of different widths. Thus, ob¬ served differences in fiber diameter could not be explained by simple

association in twos of equal size sub¬ units. Chromatin fibers are spaced irregularly with chromomere like knobs and aggregate into a thick mass of chromatin similar to chro¬ mocenters.

Digestion with 0.1 mg/ml DNase for 15 min. disrupted drastically the linear continuity of chromatin fibers leaving a heterogeneous mixture of granules and ghost fibers (Fig. 2a). Aggregates of electron dense gran¬ ules represent the remnants of con¬ densed heterochromatin. Treatment with 0.1 mg/ml RNase for 15 min. showed no marked change in chroma¬ tin fiber diameter or continuity (Fig. 2b). Both knobby appearance and aggregation of fibers into a chro¬ mocenter were still evident. Diges¬ tion with 1.0 mg/ml trypsin for 30 min. resulted in dissociation of thick fibers into thinner ones and a re¬ duction in their diameters to 40-100 A with no loss in fiber continuity (Fig. 2c). Here, however, the fibers were smooth, largely free of knobs and the chromocenter appeared less dense.

Discussion

A number of observations demon¬ strate that these interphase chroma¬ tin fibers represent the DNA — his¬ tone complex of chromosomes, and trypsin digestion removes the his¬ tone component of this complex (Ris, 1966; Bernhard and Gran- boulan, 1963 ; Bastia and Swamin- than, 1967). Therefore, it is jus¬ tifiable to define the electron dense granules and aggregates left after DNase treatment of chicken erythro¬ cyte chromatin to be primarily his¬ tone residue. Furthermore, the dis¬ sociation of thick chromatin fibers

Abdel-Hameed — Interphase Chromatin

145

witnessed in trypsin treatment in- fibers and may play a definitive role dicates that histones act as an ad- in the condensation of heteroehro- liesive promoting’ the aggregation of matin.

Figure 2. (a) A single chicken erythrocyte nucleus after DNase treatment show¬

ing ghost fibers and granules and their aggregation into one large central mass repre¬ senting the remnant of interphase chromocenter. Ghost fibers extent to what is left of the nuclear membrane on the periphry. X 3,460; (b) A single erythrocyte nucleus of chicken treated with RNase. Knobby fibers remain intact with no change in diameter. X 2,590; (c) Two erythrocyte nuclei of chicken digested with trypsin. Fibers lose their knobby appearance and dissociate into a multitude of thinner fibers. X 2.670.

146

Transactions Illinois Academy of Science

The above described osmotic shock technique used here to release chro¬ matin of intact nuclei into mono- layer protein films is much simpler and less time consuming than Gall’s Langmuir trough method, especially in the spreading and dehydration steps. It gives reproducible results and should be generally applicable to nuclear fractions of various euka¬ ryotes. Success has already been obtained with macronuclear fraction from the amicronucleate (GL) strain of the ciliate Tetrahymena pyri- fonnis and the results are summa¬ rized elsewhere (Abdel-Hameed, 1969). Interestingly, both Gall’s technique and the one described here are modifications of the water¬ spreading technique developed origi¬ nally by Kleinschmidt and his co¬ workers (Kleinschmidt and Zahn, 1959; Kleinschmidt et al., 1962) to study the fine structure of bacterial and viral genophores.

Acknowledgments

I thank Dr. M. Jollie and Dr. O. A. Schjeide for reviewing the manuscript and Miss A. Zadylak and Miss D. Molsen for laboratory assistance. Part of the present study was accomplished at Argonne Na¬ tional Laboratory through the “Faculty Research Participation Program” during the summer of 1968. This research was supported in part by Northern Illinois Uni¬ versity Grants (#02667 and 54006-46).

Literature Cited

Abdel-Hameed, F. 1969. The ultra¬ structure of macronuclear chromatin in Tetrahymena pyriformis (GL). J. Pro- tozool., ( Suppl. ) , 16:9.

Bastia, D. and M. S. Swaminthan. 1967. Ultrastructure of interphase chromo¬ somes. Exptl. Cell Res., 48:18-26. Bernhard, W. and N. Granboulan. 1963.

The fine structure of the cancer cell nucleus. Exptl. Cell Res. (Suppl.). 9:19-53.

Dupraw, E. J. 1965. The organization of nuclei and chromosomes in honeybee embryonic cells. Proc. Nat. Acad. Sci., Wash. 53:161-168.

Freifelder, D. and A. K. Kleinschmidt. 1965. Single strand breaks in duplex DNA of coliphage T7 as demonstrated

by electron microscopy. T. Mol. Biol., 14:271-278.

Gall, J. G. 1963. Chromosome fibers from an interphase nucleus. Science, 139:120-121.

- . 1966. Chromosome fibers

studied by a spreading technique. Chro¬ mosoma (Berk), 20:221-233.

Kleinschmidt, A. and R. K. Zahn. 1959. Uber Deoxyribonucleinsaure - Molekeln in Protein - Mischfilmen. Z. Natur- forschg., 146:770-779.

- , D. Lang, D. Jacherts,

and R. K. Zahn. 1962. Darstellung und Langenmessungen des Gesamten Deoxyri¬ bonucleinsaure - Inhaltes von T* - Bac- teriophagen. Biochim. Biophys. Acta., 61:857-864.

Lung, B. 1968. Whole mount electron microscopy of chromatin and membranes in bull and human sperm heads. J. Ul- trastruct. Res., 22:485-493.

Moses, M. J. 1960. Patterns of organi¬ zation in the fine structure of chromo¬ somes. Proc. 4th Int. Conf. Electron Microsc. (Berlin), 2:199-211.

Ris, H. 1966. Fine structure of Chromo¬ somes. Proc. Roy. Soc. B, 164:246-257.

- , and B. Chandler. 1963. The

ultrastructure of genetic system in pro¬ karyotes and eukaryotes. Cold Spring Harber Symp. Quant. Biol., 28:1-8.

Wetmur, J. G., N. Davidson, and J. V. Scaletti. 1966. Properties of DNA of bacteriophage NI, a DNA with re¬ versible circularity. Biochem. Biophys.

Res. Commun., 25:684-688.

Wolfe, S. L. 1965a. The fine structure of isolated metaphase chromosomes. Exptl. Cell Res., 37:45-53.

- . 1965b. The fine structure

of isolated chromosomes. J. Ultrastruct. Res., 12:104-112.

- and B. John. 1965. The

organization and ultrastructure of male meiotic chromosomes in Oncopeltus fas- ciatus. Chromosoma (Berk), 17:85-103.

Manuscript received June 4, 1970

PATHOGENESIS OF CLOSTRIDIUM BOTULINUM: IN VIVO FATE OF C. BOTULINUM TYPE A SPORES

R. BOOTH, J. B. SUZUKI AND N. GRECZ Biophysics Lab. Dept, of Biology, Illinois Institute of Technology, Chicago, Illinois 60616

Abstract. — C. botulinum Type A spores contain sufficient toxin to produce fatal botulism in experimental animals. After 4 hours, i.p. injected spores undergo con¬ version from a heat-resistant spore to a heat-sensitive cell, probably a germinated spore or vegetative cell. This conversion appears to be prerequisite for liberation of spore bound toxin, since no botulinal toxin was noted before loss of heat-resistance. Within 0.5 minutes after i.p. injection, spores enter the bloodstream, liver, spleen and kidneys. The process of spore clear¬ ance and germination appear to be retard¬ ed by Type A antitoxin binding to spores rendering them non-phagocytizable. A sig¬ nificant number of viable heat-sensitive cells of C. botulinum are found in peritoneal exudates at later stages of survival in both antitoxin-protected and non-protected ani¬ mals, indicating the capability of in vivo spore germination in the peritoneal cavity. No significant numbers of vegetative cells are found in liver, kidneys, or spleen of antitoxin protected mice. This implies that C. botulinum spores are trapped by the splanchnic mechanism, and may not be able to germinate in these organs.

Early works (Orr, 1922) on botu¬ lism pathology report C. botulinum spore dissemination into various tis¬ sues and production of toxin after oral challenge. Coleman and Meyer (1922) extended this work in dem¬ onstrating invasion of all organs of the body regardless of the mode of administration of spores. They were first to implicate toxin production in vivo from toxin-free spores.

Following intramuscular ( i . m . ) challenge, Keppie (1951) found clus¬

ters of heterophil leukocytes gath¬ ered at the site of injection which resulted in engulfment of spores and transportation away from the origi¬ nal site of injection. In vivo spore germination was rejected on the the- orectical basis that spores could not germinate in the presence of oxy¬ gen. Keppie (1951) concluded that the toxin was released from spores by phagocytic digestion.

Delayed infections by latent spores of C. botulinum has been observed in clinical cases. Assuming no re¬ current exposure to the toxin, these cases may have resulted from the ingestion of foods heated sufficiently to destroy toxin, but not heat-resist¬ ant spores. Therefore, ingestion of C. botulinum spores as an etiological agent in botulism food poisoning must be considered.

Two possibilities have been offered (Grecz and Lin, 1967) to explain spore toxicity: 1. in vivo spore ger¬ mination and subsequent release of toxin from vegetative cells, and 2. degradation of the spore releasing its bound toxin.

The purpose of this paper is to demonstrate that C. botulinum spore do, in fact, germinate in vivo , and to define in vivo loci of mice which are conducive to this germination.

[147]

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Transactions Illinois Academy of Science

Materials and Methods

Culture methods: Clostridium botu- linum Type A strain 33A was ob¬ tained from Dr. W. E. Perkins, Na¬ tional Canners Association, Berkeley, California. The culture was grown at 30 C in 5% Trypticase (BBL), 0.5% peptone (Difco) and 0.1% sodium thioglycolate. Within 6 days, abundant sporulation had occurred, at which time, the spores were har¬ vested in a refrigerated Sorvall RC-2 continuous centrifugation system and cleaned with trypsin and ly¬ sozyme by method of Grecz, et al. (1962). The spores were washed twice, resuspended in 0.87% NaCl and stored at 4 C until used. Heat-shocking procedure : The spore inoculum was heated in a screw-cap tube for 15 minutes at 80 C dena¬ turing any free botulinal toxin in the medium. C. botulinum spores are able to survive this treatment. Colony counts : The number of vi¬ able organisms of C. botulinum were determined by subculture in Wynne’s broth (Wynne, et al 1955) plus 0.75% agar. One ml portions of serial dilutions were transferred to oval flat tubes and melted sterile Wynne’s agar was added. To achieve anaerobiosis, an additional layer (2 cm.) of sterile Wynne’s agar was poured. The tubes were plugged with foam rubber stoppers and in¬ cubated at 30 C. Colonies were count¬ ed after 96 hours.

Mice : White Swiss mice raised for 10 generations in a closed colony at the Illinois Institute of Technology were utilized in all experiments. They were fed and watered ad libi¬ tum , and attained a weight of 25 grams before experimentation.

Injections : Intraperitoneal (i.p.) injections into mice were made with 26 gauge, 2.5 ml disposable syringes. Preparation of Exudates for Viabil¬ ity Analysis : Mice were injected i.p. with 2 x 10s spores. At selected time intervals, the peritoneal cavity was washed with 1 ml of sterile 0.87% saline, and exudates with¬ draw using a 26 gauge syringe. The peritoneal exudate was diluted 1 :10 with sterile distilled water. Serial dilutions in distilled water for viable cell count in Wynne ’s agar were per¬ formed as described above. Preparation of Tissues for Viability Analysis : Following removal of peri¬ toneal exudates from mice, the ani¬ mals were etherized and the peri¬ toneal cavity surgically opened. The body cavity was rinsed with sterile saline, and the liver, kidneys and spleen removed.

The organs were placed in sterile plastic centrifuge tubes and washed with sterile saline by agitation on a vortex mixer. Washing was contin¬ ued until the saline supernates be¬ came clear. Each organ, regardless of volume, was placed in 20 ml of sterile saline in a second sterile plas¬ tic centrifuge tube, and sonicated (Branson Sonifier, Model S-125, 8 amps) for 2-4 minutes to rupture the cells and release all spores.

The tubes were centrifuged (300 x g, 1 hour), and the supernatant discarded. Appropriate decimal di¬ lutions of the pellet were analyzed for the number of viable spores in Wynne’s agar.

Examination of Blood for Viable Spores and Vegetative Cells : Blood was removed from the mouse by two methods: (i) the animal was ether¬ ized and the abdominal and pleural

Booth et al. — C. botulinum Pathogenesis

149

cavities were surgically opened using sterile techniques. A 26 gauge, 2.5 ml syringe was inserted into the heart and the blood was withdrawn ; (ii) a direct cardiac puncture was made into a mouse slightly stunned by ether. Decimal dilutions of the blood were made and examined for the number of viable organisms in AVynne ’s agar.

Results

Peritoneal Cavity : After i.p. injec¬ tion of 2 x 108 heat-shocked spores of C. botulinum Type A, onl}7- 6.6 x 105 heat-resistant spores could be recovered from the peritoneal cavity at the start of the experiment ( Table 1). This apparent 300 fold reduc¬ tion in number of spores was due to : (i) dilution of spores by body fluids ; or (ii) rapid distribution of the

spores throughout the animal body.

The first line of Table 1 shows that the intraperitoneal cavity of control mice receiving no spore inoculum contained some spores (6.0 x 102) as well as some heat-sensitive cells (4.4 x 103). This relatively low lev¬ el of contamination was considered as “ background’’ flora. Examina¬ tion of Table 1 showed that the num¬ ber of spores recovered from the ani¬ mal body after initial i.p. injection gradually but consistently declined so that by 48 hours the number of spores recovered was approximately 100 fold lower than the initial level. Furthermore, at 48 hours, the num¬ ber of spores recovered was only slightly higher than ‘'background” spore load in control mice.

Incubation beyond 3 hours rapid¬ ly increased the number of lieat-

Table 1. — Number of viable spores recovered from the peritoneal cavity of mice after intraperitoneal injection of 2 x 108 heat-shocked spores of Clostridium botulinum 33A.

Number of Viable Organisms Recovered

Time After Intraperitoneal Injection Hours

Number of Animals Challenged

Heat-Shocked

(Spores)

Heat Sensitive Cells (Total Minus Spores)

Control® .

3

6.0 x 102

4.4 x 103

0 .

3

6.6 x 105

4 x 103

1 .

2

5. Ox 105

4 x 104

2 .

3

1.0 x 105

1.0 x 103

3 .

3

2.0 x 105

5 x 104

4 .

3

1.0 x 105

2 x 105

8 .

3

2.0 x 104

1.5 x 106

24 .

3

2.0 x 104

4.0 x 104

1.8 x 105

5.0 x 104

36 .

1

2.0 x 104

4.3 x 105

1

1.0 x 104

1.0 x 104

1

5.0 x 102

2.5 x 103

1

1.0 x 103

3.0 x 103

48b .

2

7.0 x 103

5.0 x 105

1

1.0 x 103

2.0 x 10s

1

3.0 x 103

1.0 x 105

1

2.0 x 103

5.0 x 103

a/Control mice received 1 ml sterile saline but no spores.

••/Recovered from surviving animals; only 10% of the injected animals survived 48 hrs.

150

Transactions Illinois Academy of Science

sensitive cells and reached a plateau of approximately 2 x 105 cells which seemed to be relatively stable be¬ tween 8 to 48 hours (the upper limit of this experiment). Within the precision of these experiments, the number of heat-sensitive (germinat¬ ed) cells was essentially equivalent to the initial number of spores which could be recovered from the peri¬ toneal cavity of the mouse during the first 8 hours. Thus, these results suggest that after injection into the mouse, heat-resistant spores were es¬ sentially all converted into heat-sen¬ sitive (probably germinated) forms

within 4-8 hours. The heat sensitive forms appeared not to be destroyed to a detectable degree in the mouse body for up to 48 hours.

The cells recovered from the peri¬ toneal cavity of injected mice were shown to produce specific Type A toxin as determined by mouse tox¬ icity tests of subculture. Control mice in which the animals were chal¬ lenged with 1 ml of sterile saline without spores contained low levels of viable cells. The organisms re¬ covered from the controls were non- toxic.

It is important to note that mice

Figure 1. Rate of death of mice in¬ jected with spores of C. botulinum as com-

# " # Mice injected i.p. with 2 x

10s heat-shocked spores con¬ taining 10 LD.-,o of type A toxin.

pared with mice injected with free type A botulinum toxin.

O . O Mice injected i.p. with a di¬

lution of pure type A botu- li uim toxin.

Booth ct at. — C. botulinmn Pathogenesis

151

injected with 2 x 10s spores (con¬ taining the equivalent of 10 LD-„ botulinal toxin by mice assay) did not show any typical symptoms of botulism for the first 8 hours. Fifty- percent of the animals died 32 hours after injection (Figure 1). Nor¬ mally, mice injected with 10 LD5(1 of botulinum toxin expire within 8-18 hours, with 50% expiring with¬ in 10 hours. Presumably, the in- creased expiration time with spores is needed for in vivo spore germina¬ tion. Millipore Filtrate (0.22 of the spore suspension were not le¬ thal to any mice injected.

These observations suggest that germination of C. botulinum spores occurs previous to toxin release in vivo.

Spores in the Blood : Table 2 demon¬

strates that heat-resistant spores penetrate into the blood stream with¬ in 0.5 minutes after i.p. injection. It can be seen that the cells remain heat-resistant for at least 70 min¬ utes, a fact which is in accordance with lack of spore germination or loss of heat-resistance during the first 3 hours as pointed out in Table 1. The number of spores entering the blood appears to be somewhat lower than the number of spores recovered from peritoneal exudate during compara¬ ble time intervals. However, the

data definitely establish the fact that %/

spores penetrate extremely rapidly

into the bloodstream. Background

flora bacteria, upon penetration into

the bloodstream are effectively de-

*/

stroyed by phagocytes in the blood¬ stream.

Table 2.- — Number of Viable Spores in the Blood of Mice Injected Intraperitoneally with 2 x lO* Spores.

Time of Withdrawal (min.)

Method of Withdrawal*

Number of Heat-Shocked Spores (per ml.)

Number of Non-Heat- Shocked Spores (per ml.)

0 .

A

0

0

.5 .

A

1 x 104

1 x 104

15 .

B

3 x 105

5 x 104

20 .

A

6 x 103

6 x 103

A

3 x 102

3 x 104

30 .

B

7 x 104

1 x 103

B

2.5 x 104

5 x 103

40 .

A

1 x 103

1 x 103

A

2 x 103

2 x 103

60 .

A

2.1 x 103

2 x 103

70 .

A

7 x 102

5 x 10s

* Method A — direct cardiac puncture while animal etherized. Method B — animal sacrificed, body opened, cardiac puncture.

152

Transactions Illinois Academy of Science

Spores in Liver, Kidney and Spleen : Table 3 shows that spores of C. botu¬ linum 33A injected i.p. are rapidly disseminated to the liver, kidneys and spleen between 0.5 and 4 hours. The spores persist in the liver and kidneys at a relatively constant level up to 36 hours at which time a rapid decrease in the number of viable or¬ ganisms occurred. In the spleen, spore clearance initiated at 8 hours and continued for 24-48 hours. The number of heat-sensitive (germi¬ nated) spores or vegetative cells was usually approximately 5 fold higher than the number of heat-resistant spores.

As with the peritoneal exudate, control colonies from organ prepa¬ rations were not toxic to mice. The Colonies in 10 experimental plates proved to be toxic as determined by

mice i.p. assay (antitoxin-protected mice survived). Moreover, organs from healthy animals, ground and in¬ jected into mice, did not cause ill effects. These controls substantiate the presence of C. botulinum spores in the organs.

RES Clearance of Spores in Mice Protected by Antitoxin : In order to obviate the effect of spore toxin, a series of samples similar to those in the preceding experiment were ana¬ lyzed, with the exception that 0.1 ml of antitoxin sufficient to protect the animal from death was administered with each spore inoculum. In this way, it was possible to evaluate the survival of spores in vivo for extend¬ ed periods of time.

In the peritoneal exudate of pas¬ sively immunized mice, appearance of a heat-sensitive element could not

Table 3.- — Number of viable cells recovered from organs of mice after intraperi- toneal injection of 2 x 108 heat-shocked spores of Clostridium botulinum 33A. (Mice received no antitoxin).

Viable Cells Recovered From Organs4

Time After Intraperitoneal Injection (hrs.)

Livers

Kidneys

Spleens

Spores

Heat

Sensitive

Cells

Spores

Heat

Sensitive

Cells

Spores

Heat

Sensitive

Cells

0 (controls) .

1 x 102

2 x 104

3 x 101

1.7 x 102

3 x 101

1.7 x 102

1/2 .

2 x 103

8 x 104

1 x 103

1 x 104

3 x 103

2.7 x 104

1 .

1 x 104

1 x 106

3 x 104

2 x 103

8 x 103

7.2 x 104

2 .

5 x 104

3 x 104

2 x 103

3 x 104

3 x 104

1 .7 x 105

3 .

7 x 104

1 x 105

2 x 103

2.8 x 104

2 x 104

1.8 x 105

4 .

7 x 104

2 x 105

2 x 103

2.8 x 104

7 x 104

1.3 x 106

8 .

5 x 104

1.6 x 106

1 x 104

2 x 104

1 x 105

1 x 105

18 .

7 x 104

2.3 x 105

3 x 104

7 x 104

24 .

1 x 105

3 x 105

3 x 104

7 x 104

3 x 103

2.7 x 104

36 .

1 x 105

2 x 105

3 x 104

2 x 104

1 x 103

8 x 103

48 .

2 x 103

5 x 104

1 x 103

2 x 103

2 x 103

1 . 5 x 103

) Control animals received 1 ml sterile saline without spores Averages of 4 animals.

Booth et al. — C. botulinum Pathogenesis

153

be observed as in mice not passively immunized, (Figure 2). The num¬ ber of spores decreased slightly dur¬

ing the first 5 days, but dropped approximately 100 fold between the fifth and seventh day. Between one

DAYS

Figure 2. Number of Viable Spores in the Peritoneal Cavity of Mice Injected with 2 x 10s Heat-shocked Spores of C. botulinum 33 A.

Spores: 1 ml of cells sur¬ viving 80C for 10 min. in type A antitoxin protected mice.

if . ★ Spores: 1 ml of cells sur¬

viving 80C for 10 min. without antitoxin.

O Heat-sensitive cells: Cells killed by 80C for 10 min. in type A antitoxin pro¬ tected mice.

Heat-sensitive cells: Cells killed by 80C for 10 min. without antitoxin.

154

Transactions Illinois Academy of Science

week and 3 weeks the numbers of spores continued to decrease further. In the presence of antitoxin, heat- sensitive C. botulinum cells did not appear in numbers greater than nor¬ mal background contamination, i.e. approximately 4 x KT, indicating that destruction of spores in vivo after germination must have been extremely rapid.

In the liver, spleen and kidneys, no situation was found which could be attributed to the presence of vege¬ tative cells in presence of antitoxin, since heat-resistant and heat-sensi¬ tive counts were always essentially equal (Table 4). There was gradual disappearance of spores from all these organs during the three weeks. Controls injected with antitoxin alone, contained 50 to 1000 spores, and 1000 to 10,000 heat-sensitive

cells, all of which did not produce toxin in subculture.

Discussion

The significant finding emerging from these studies is the fact that spores injected i.p. — with or with¬ out antitoxin — are disseminated within seconds or minutes into the blood, liver, spleen and kidneys of the experimental animal. Subse¬ quent clearance of spores from the RES is very gradual.

The mechanism of intoxication of the animals from spore toxin appears to depend on conversion of the heat- resistant spores to heat-sensitive forms ; perhaps germinated spores or vegetative cells. The heat-sensitive cells appear at 4 hours and may per¬ sist for over 48 hours after i.p. injec-

Table 4. — Number of viable cells mice after intraperitoneal injection of 2 x 33A.

recovered from organs of antitoxin protected 10* heat-shock spores of Clostridium botulinum

Viable Cells Recovered From Organs

Time After Intraperitoneal Injection

Livers

Kidneys

Spleens

Sporesa

Total

Spores

Total

Spores

Total

0 .

1 x 102

2 x 103

3 x 101

3 x 102

3 x 101

2 x 102

20 hours .

2 x 10°

1 x 106

7 x 104

5 x 104

6 x 105

5 x 105

42 hours .

2 x 106

2 x 106

7 x 105

2 x 105

1 x 106

6 x 105

48 hours .

1 x 106

3 x 106

2 x 104

1 x 104

4 x 105

2 x 105

67 hours .

9 x 105

5 x 105

7 x 104

3 x 104

3 x 105

1 x 105

120 hours .

1 x 106

7 x 105

2 x 104

3 x 104

2 x 105

1 x 105

1 week .

9 x 105

1 x 106

1 x 105

1 x 106

2 x 105

1 x 106

2 weeks .

1 x 105

1 x 105

1 x 104

9 x 103

2 x 104

2 x 104

3 weeks .

2 x 104

2 x 104

1 x 104

1 x 104

5 x 103

5 x 103

5 x 105

5 x 105

1 x 104

2 x 104

5 x 104

5 x 104

a/Spore = cells surviving heating at 80 C for 10 minutes.

No germination, no appearance of heat-sensitive forms.

Control mice inoculated with 0.1 ml antitoxin but no spores contained 50 to 1000 spores and 1000 to 10,000 heat-sensitive cells.

Booth et al. — C. botulinum Pathogenesis

155

tion. Symptoms of botulism intoxi¬ cation are observed after time intervals necessary for loss of heat- resistance by injected spores.

The presence of botulinal antitoxin A appears to depress normal spore clearance levels in vivo by a factor of 5-10x. By a technique developed by this laboratory (Suzuki, et al., 1971a), botulinal toxin was found not to affect phagocytic clearance (Suzuki, et al., 1971b) which is in agreement with other investigators (Freeman, 1961). We speculate that delayed disappearance of spores in vivo when antitoxin is administered may be due to antitoxin binding to spores rendering them dormant. This seems plausible as a host defense me¬ chanism against botulism pathogene¬ sis since Suzuki, et al. (1970, 1971c) reported the PMN leukocyte as req¬ uisite for C. botulinum spore germi¬ nation and toxin release. Staining techniques of phagocytized C. botu¬ linum spores have verified this rela¬ tionship (Booth, et al 1971).

Acknowledgement

This study was supported by PHS Grant FD-00095 from the Food and Drug Admin¬ istration and PHS Career Development Award 5-K3-3-AI-21,763.

Literature Cited

Booth, R., J. B. Suzuki, and N. Grecz. 1970. Germination of Clostridium botu¬ linum 33A spores within the animal

body. Bact. Proc. G127, p. 34.

- . 1971. Sequential use of

Wright’s and Ziehl-Neelsen’s Stains for Demonstrating Phagocytosis of Bacterial Spores. Stain Technol. 46:23-26.

Coleman, G. E. and K. F. Meyer. 1922. Some observations on the pathogenicity of B. botulinus x. J. Infect. Dis. 31 : 622-649.

Freeman, N. L. 1961. Phagocytosis of staphylococci by mouse leucocytes in the presence of botulinum toxin. J. Bac- eriol. 81: 156-157.

Grecz, N., A. Anellis, and M. D. Schneider. 1962. Procedure for clean¬ ing of Clostridium botulinum spores. J. Bacteriol. 84: 552-558.

- , and C. A. Lin. 1967.

Properties of heat resistant toxin in spores of Clostridium botulinium 33 A: 302- 322. In M. Ingram and T. A. Roberts (eds.) “Botulism 1966”. IX Interna¬ tional Congress of Microbiology, Moscow, July 20-22, 1966. Chapman and Hall Ltd., London.

Keppie, J. 1951. The pathogenicity of the spores of Clostridium botulinum. J. Hyg. 49: 36-45.

Orr, P. F. 1922. The pathogenicity of Bacillus botulinus. J. Infect. Dis. 30: 118-127.

Suzuki, J. B., R. Booth, and N. Grecz, 1970. Pathogenesis of Clostridium botu¬ linum Type A: Release of toxin from C. botulinum spores in vitro by leuko¬ cytes. Res. Comm. Chem. Path. &

Pharm. 1 : 691-71 1.

- . 1971a. Evaluation of

phagocytic activity using labeled bac¬ teria. J. Infect. Dis. 123:93-96.

- . 1971b. Effect of botu¬ linal toxin on phagocytic index. (in press) .

. 1971c. In vivo and in vitro release of 4CCa from spores of Clostridium botulinum Type A as further evidence for spore germination. Res. Comm. Pathol. & Pharm., 2:16-23.

Manuscript received January 25, 1971

THREE IRON SULFATE MINERALS FROM COAL MINE REFUSE DUMPS IN PERRY COUNTY, ILLINOIS

DENNIS GRUNER AND WILLIAM C. HOOD

Department of Geology,

Southern Illinois University, Carhondale , Illinois 62901

Abstract. — Three iron sulfate minerals have been found on coal mine refuse dumps in Perry County, Illinois. Szomol- nokite (FeSOcHoO) usually occurs on oxidizing pyrite on the dump surface, ro- zenite (FeSCVTHoO) is found beneath the surface and along some watercourses, and melanterite (FeSOW^O) appears to be restricted to moist areas. The occurrences indicate the hydration state of the ferrous sulfate is at least partly dependent upon relative humidity of the microenvironment.

Pyrite is a common accessory min¬ eral in the coal deposits of southern Illinois and elsewhere. During the benefication of the coal, pyrite, as well as clay, shale, and other impuri¬ ties, is discarded and piled up in refuse dumps. Oxidation of pyrite in the upper portion of these dumps has resulted in a variety of products, three of which are described in this paper. This study was conducted on dumps located in Perry County, Illi¬ nois, but conditions similar to those found in this area are so common throughout coal mining regions of midcontinent United States and elsewhere that the minerals described herein are probably of widespread occurrence.

Methods

The samples used in this study were collected from two coal mine refuse dumps located two miles southwest of DuQuoin and five miles

south of Pinckneyville, Perry Coun¬ ty, Illinois. Collection of samples was accomplished during the months of September and October, 1969, in fairly dry conditions. Approximate¬ ly 100 specimens were collected from dry runoff ditches, along flowing streams and seeps, and from fresh bulldozer cuts in the refuse dumps. Samples were grouped in the labora¬ tory on the basis of observable prop¬ erties. They were then hand cleaned under a microscope to remove impur¬ ities in order to insure relatively pure specimens for X-ray identifica¬ tion. The purified minerals were finely ground and identified by X-ray powder diffraction techniques.

Results

X-ray diffraction study reveals the occurence of three hydrated iron sul¬ fate minerals: szomolnokite (Fe04. •H20) ; rozenite (FeS04‘4H20) ; and melanterite (FeSOyTFUO) . About 15% of the fifty samples identified by X-ray diffraction were melanter¬ ite, 30% rozenite, and 55% szomol¬ nokite (percentages do not neces¬ sarily reflect relative abundance on the refuse dumps, but rather propor¬ tion of X-ray test samples ; however, the abundance of the various miner¬ als in the test runs are approximate¬ ly proportional to their abundance

[156]

Gruner and Hood — Sulfate Minerals, Perry County

157

in the total samples collected.) These three minerals have sufficiently dif¬ ferent physical properties so that they can generally be identified in the field without the aid of sophis¬ ticated equipment. A brief descrip¬ tion of each mineral follows.

Szomolnokite. Szomolnokite oc¬ curs as a white powdery coating on pyrite and occasionally over the en¬ tire surface of an area where oxidiz¬ ing pyrite is abundant. Thus far we have found it only on material exposed directly at the surface.

It is distinguished by its occur¬ rence as a white powder on pyrite, its hardness of 2.5 (determined only with considerable difficulty), and its slow rate of dissolution in water. It is reported that a brown residue is left when the mineral is dissolved in water (Palache, et. ah, 1951), but we have not found this to be an im¬ portant characteristic. It does not have a cleavage. The specific gravity is about 3.05 (Winchell and Win- chell, 1964), but its mode of occur¬ rence generally makes an estimation of specific gravity impossible.

Rozenite. Rozenite is rather com¬ mon is Perry County. The best oc¬ currences are a few inches beneath the surface of the spoil piles, where considerable masses of granular crys¬ tals can occasionally be found. It also occurs along the sides of some small streams that drain the spoil piles, but in such locations it usu¬ ally turns to a crumbly white ma¬ terial if the water dries up.

Rozenate has a hardness of 2.5, which is the same as szomolnokite but clearly harder than melanterite. The specific gravity is about 2.3. Winchell and Winchell (1964) re¬ port artificial rozenite has one direc¬

tion of good cleavage, but the well developed conchoidal fracture of the Perry County material often makes the cleavage difficult to discern. Most samples used in this study were blue to blue-green, but occasionallv colorless or white material was found. Unaltered specimens are transparent to translucent and have a vitreous luster ; partially de¬ hydrated specimens are translucent to opaque and have a dull luster. The mineral is readily soluble in water and gives a metallic, astringent taste.

Melanterite. This mineral is gen¬ erally found as a crust on rocks and mud along the sides of flowing drain¬ age ditches, usually within a few inches of water. It occasionally is found as small clumps of white ra¬ diating crystals.

Melanterite is transparent to translucent and has a vitreous luster. The color ranges from colorless to green or blue-green. Both its hard¬ ness (2.0) and its specific gravity (1.90) are considerably less than those of rozenite, with which it can easily be confused. Like rozenite, it dissolves easily in water and has a metallic, astringent taste. Its three directions of cleavage are not easily seen in most specimens. The most useful field test to distinguish melan¬ terite from rozenite is the hardness.

Discussion

Collection precautions. In col¬ lecting samples of these minerals for research or mineral collections, care must be taken to preserve them in their natural hydration states. It is suggested that they be placed in airtight containers and tightly sealed

158

Transactions Illinois Academy of Science

immediately upon collection, because only a few hours exposure to dry air will cause drastic changes in appear¬ ances. Rozenite appears to be par¬ ticularly susceptible to alteration, for it will change from a glassy, transparent blue or blue-green ma¬ terial to a dull, opaque, white, crumbly material overnight if ex¬ posed to dry air. On the other hand, we have successfully kept one sam¬ ple for over a year with no sign of alteration by storing it in a tightly sealed polyethylene bottle.

Another problem in the handling and storage of these minerals is their decidedly acid reactions. Dissolving appreciable quantities of the miner¬ als in water will lower the pH to the range of 2 to 4. Paper or aluminum foil left in contact with the minerals, particularly in a moist atmosphere, will rapidly decompose.

Stability relations. The occurrence of melanterite and rozenite in Perry County does not seem to entirely conform to the present state of knowledge of the system ferrous sul¬ fate-water-sulphuric acid. Experi¬ mental work by Cameron (1930) in¬ dicates that melanterite should be the stable ferrous sulfate phase cry- stalizing from solutions with less than 28 percent sulphuric acid at 25° C. Ehlers and Stiles (1965) proved that melanterite can be re¬ versibly dehydrated to rozenite un¬ der relative humidities of less than 70 to 80 percent at temperatures that would be found on the surfaces of the refuse dumps. Taken together, these two studies suggest that the ferrous sulfate phase crystallizing from coal mine drainage water ought to be

melanterite and subsequent exposure to low humidity air could cause dehy¬ dration to rozenite. The close as¬ sociation of melanterite with water is in agreement with this conclusion. However, we have repeatedly ob¬ tained rozenite directly by evapora¬ tion of a saturated solution of ro¬ zenite in water under conditions that should have given, according to Cam¬ eron (1930), melanterite. Further¬ more, the subsurface occurrence of rozenite in moist soil suggests that it can exist under conditions of high relative humidity. This is in dis¬ agreement with the earlier studies and suggests the natural system dif¬ fers in some significant way from the artificial one. Work is being initi¬ ated in this laboratory in an attempt to discover the factors that stabilize rozenite in the coal mine refuse dumps of Perry County.

Acknowledgments

The authors would like to thank the Truax-Traer Coal Company for allowing us to collect samples on its property. John Ramsey, project manager of the Truax- Traer water pollution control experimental project, was especially helpful in suggest¬ ing collecting localities and in donating some samples. Paul Robinson read the manuscript and suggested improvements.

Literature Cited

Cameron, F. K. 1930. The solubility of ferrous sulphate. J. Phys. Chem. 34: 692-710.

Ehlers, E. H., and D. V. Stiles. 1965. Melanterite-rozenite equilibrium. Am. Mineral. 50: 1457-1461.

Palache, C., H. Berman and C. Frondel. 1951. Dana’s System of Mineralogy. 7th ed., Vol. II. John Wiley and Sons, New York xi + 439 pp.

Manuscript received April 27, 1970

THE BEHAVIOR OF IRON IN PEORIA LAKE

WUN-CHENG WANG AND RALPH L. EVANS Water Quality Section, Illinois State Water Survey, Peoria, Illinois

Abstract. — Iron in Peoria Lake can be differentiated into several fractions, of which the great majority was found to be particulate Fe(III). The particulate Fe (III) possess several characteristics. Its concentration is much higher in the upper reaches of the lake than in the down¬ stream sector; it can be correlated with water turbidity; there is a significant cor¬ relation between particulate Fe(III), par¬ ticulate Fe(II), particulate silica, and par¬ ticulate phosphate. The dissolved Fe(III) concentration is in excess of its solubility, suggesting that it is not in a complete soluble form.

Aqueous solutions of iron have been intensively studied during the past decade. Many investigations have been concerned with the chemi¬ cal behavior of iron as related to thermodynamic and kinetic models (Hem and Cropper, 1959; Hem, 1960 ; Stumm and Lee, 1960 ; Morgan and Stumm, 1964; Ghosh, O’Connor, et. al., 1966; Larson, 1967). Nu¬ merous as such studies have been, most of them have dealt with dis¬ tilled water sj^stems. The results obtained from such systems can not, without modification, be applied to a natural water system. For exam¬ ple most iron studies have been per¬ formed under abiotic laboratory con¬ ditions. Such studies though in¬ formative are not substantive; bio¬ logical activity does affect the iron behavior in natural water, as sug¬ gested by Lee and Hoadley (1967). Furthermore, Morgan and Stumm (1964) have shown, in distilled water

studies, that iron most likely plays a role in surface chemical reactions. As natural water contains suspended solid such as detritus, silt, and clay, it is conceivable that the surface phe¬ nomenon may be even more impor¬ tant in the natural water system. In an effort to gain some insight into the behavior of iron in a natural water environment the observations presented herein were made, as a part of a limnological study of Peo¬ ria Lake.

Procedure

Peoria Lake, is fundamentally a wide basin of the Illinois River (Fig¬ ure 1). It has been channelized for navigation and is located in one of a series of eight pools extending from the river’s confluence with the Mis¬ sissippi River. At normal pool stage the lake is about 13 miles long and has an average width of 1.5 miles; the maximum depth is 5 meters with a considerable portion of water depth less than 1 meter. During the study period, the residence time within the lake ranged from 2 to 6 days.

Five transects and nine stations were established on the lake (Fig¬ ure 1). On four of the transects two stations were assigned ; one was representative of the channel area and the other the shallower area of the lake. A single station was

[159]

160

Transactions Illinois Academy of Science

2.51

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Figure 1. Various iron fractions in Peoria Lake.

Wang and Evans

Behavior of Iron

161

selected in the “Narrows”, the out¬ let of the lake.

Water samples were collected at the nine stations at a depth of 1 meter using* a Kemmerer sampler. Collection was made at one to two weeks intervals. All water samples were transferred to glass bottles which were prewashed with acid.

Analytical Methods

The turbidities of all samples were determined immediately upon deliv¬ ery to the laboratory. The method used closely parallel the Jackson candle method (American Public Health Association, 1965). Also up¬ on delivery a portion of each sample was filtered through an 0.45 micron membrane filter.

The filtrate and unfiltered portions of each sample were separately ana¬ lyzed for Fe(II) and total iron by using* the phenanthroline method

(American Public Health Associa¬ tion, 1965). In this manner the con¬ centration of the various fractions of iron were determined, i.e., dis¬ solved Fe(III) and Fe(II) and par¬ ticulate Fe(III) and Fe(II). All analyses were performed within 48 hours after the collection of samples.

Dissolved oxygen, temperature, and pH were determined in the field. Dissolved oxygen and temperature were determined by an oxygen ana¬ lyzer manufactured by Precision Si- entific Company, and pH was deter¬ mined by a Beckman model N pH meter.

Results and Discussion Distribution

Some characteristics of Peoria Lake water are shown in Table 1. In general, the water is turbid and rich in the bicarbonates of calcium and magnesium. Nutrient levels

Table 1. — Chemical Characteristics of Peoria Lake

Range

Mean**

Temperature, °C .

5.0 —

27.3

19.5

Turbidity, JTU .

28.0

296.0

115.0

pH .

7.57 -

8.69

8.19

Dissolved Oxygen, mg/1 .

1.4 —

15.3

5.6

Alkalinity*, mg/1 .

136.0 —

213.0

165.0

Hardness*, mg/1 .

215.0 —

324.0

268.0

Iron (total), mg/1 .

0.69 —

13.01

3.21

Ferrous .

0.16 —

1.89

0.58

Ferric .

0.52 —

11.12

2.63

Fluoride, mg/1 .

0.17 —

2.06

1.08

Silica (total), mg/1 .

1.96 —

14.80

6.10

Nitrogen (total), mg/1 .

3.88 —

14.98

8.85

Nitrate (NO3-N) .

1.65 —

11.12

4.33

Ammonia (NO3-N) .

0 —

5.45

1.15

Organic-N .

0.64 —

9.84

3.37

Phosphorus (total), mg/1 .

0.47 —

3.02

1.13

Orthophosphate-P .

0.25 —

2.30

0.84

Polyphosphate-P .

0 —

0.67

0.15

Organic Phosphate-P .

0 —

0.58

0.14

* expressed as CaCOt ** based on 225 samples

162

Transactions Illinois Academy of Science

are quite high and upstream wastes discharges are reflected in the rela¬ tively low dissolved oxygen content.

The distribution of various iron fractions is shown in Figure 1. Of the four iron fractions, particulate Fe(III) was dominant and consti¬ tuted over 70 percent of total iron in the lake waters. The next abun¬ dant fraction was particulate Fe(II), followed by dissolved Fe(III), and dissolved Fe(II), in that order.

An attempt was made to determine the fluctuation of particulate Fe- (III) from station to station. A two-way analysis of variance was made. The result showed that par¬ ticulate Fe(III) concentration was significantly different from station to station, i.e., it was not uniformly distributed in the whole lake. Fur¬ ther attempts were made to group the nine stations into three regions; stations 1 and 3, stations 2, 4, 5, and 6, and stations 7, 8, and 9. The analysis of variance was again made in three regions separately. The re¬ sults showed that there was no long¬ er significant variation of iron con¬ centration within each region. Fig¬ ure I depicts the results for the three regions.

Particulate Fe(III) was highest at station 1 and 3, ranging from 3.08 to 3.15 mg/1. The intermedi¬ ate zone was at station 2, 4, 5, and 6, ranging from 2.51 to 2.90 mg/1. The lowest concentrations were at stations 7, 8, and 9, ranging from 1.76 to 1.93 mg/1. This distribution pattern is the same as that observed for turbidity in the lake (Wang and Brabec, 1969).

It should be noted that the per¬ centage of particulate Fe(III), with regard to the total, ranged from 73

to 80. The highest level was at sta¬ tion 2 with a gradual decrease to the lowest level at station 9. This was apparently due to a greater loss of particulate Fe(III) through pre¬ cipitation along in the water course compared with that of other iron fractions.

If station 2 is considered repre¬ sentative of the inlet and station 9 the outlet of the lake the iron budget can be computed. Figure 2 depicts the results in terms of concentration and load. The positive iron balance indicates that the iron input is great¬ er than the output with the conse¬ quence accumulation of iron within the lake.

The observed dissolved Fe(III) concentration averaged 0.20 mg/1 which is ten times higher than the solubility of this fraction as cited in the literature (Stumm and Lee, 1960). This suggests that the pre¬ ponderance of dissolved Fe(III) is not insoluble form but exist pos¬ sibly as particles of less than 0.45 micron diameter. Shapiro (1964) reported the existence of Fe(OH)3 in the form of a precipitate and pep¬ tized sol.

Iron and Turbidity

Since turbidity can be regarded as an index of particulate matter, an attempt was made to correlate par¬ ticulate iron and turibidty. Figures 3 and 4 show the relationship be¬ tween turbidity and Fe(III) and turbidity and Fe(II), respectively. Regardless of location within the lake, particulate Fe(III) and Fe- (II) were significantly related with turbidity. A similar analysis with dissolved Fe(III) and Fe(II) did not reveal a similar relationship.

Wang and Evans — Behavior of Iron

163

The precise structure of particu¬ late iron on the surface of particu¬ late matter is unknown. As previ¬ ously mentioned it can be in the form of a precipitate granule, film, or peptized sol (Shapiro, 1964). It would seem likely that a strong link¬ age exist between iron and particu¬ late matter. Iron, being an electro- phylic substance, is naturally in¬ clined to attach to clay, a nucleophy- lic substance, the “backbone” of particulate matter in water.

Dissolved iron, mentioned earlier as not being truly soluble in Peoria Lake may be in the form of neutral salt or an electron-rich form which

renders it free from electrostatic attraction to clay minerals or organic detritus.

Iron and Other Elements

A significant relationship between turbidity and particulate iron was observed. A similar relationship be¬ tween turbidity and particulate sili¬ ca was also found (Wang and Evans, 1969). It is thus logical to expect that particulate iron is also signifi¬ cantly correlated with particulate silica and phosphate. These rela¬ tionships are shown in Figures 5 and 6 and summarized in Table 2.

The overall average concentrations

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Transactions Illinois Academy of Science

Fe(II) CONCENTRATION, mg/liter

Wang and Evans — Behavior of Iron

165

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Transactions Illinois Academy of Science

Table 2. — Correlation Coefficients of Various Parameters in Peoria Lake

Part.

Fe(III)

Part.

Fe(II)

Part.

Si04

Part.

P04

Turbidity

Part. Fe(III) .

0.587**

0.704**

0.850**

0.854**

Part. Fe(II) . .

0.587**

0.218

0.257

0.751**

Part. SKL .

0.704**

0.218

0.444*

0.454*

Part. PO4 ....

0.850**

0.257

0.444*

0.648**

Turbidity .

0.859**

0.751**

0.454*

0.648**

* = 95% significance level ** = 99% significance level

of particulate iron, silica, and phos¬ phorus were 2.48, 1.80, and 0.42 mg/1, respectively. These values rep¬ resent a molar ratio of 3.15, 2.15 and 1 in the same order. In other words, on particulate matter the computed sum of silica and phos¬ phorus molecules was exactly same as the iron molecules. The result suggests a stoichiometric relationship among these three constituents.

The particulate matter in Peoria Lake is believed to be mainly silt and clay particles. These particles

carry negative charges principally due to isomor phous substitution. In a water environment, these particles may associate with various elements through physico-chemical forces. Of the three constituents — particulate Fe(III), silica and phosphorus — particulate Fe(III) is the most like¬ ly one to attach to particulate mat¬ ter. Silica and phosphorus may then link with particulate matter through the iron “bridge’/ a propounded mechanism in soil chemistry (Evans and Russell, 1959).

1

0

9

8

7

6

5

k

3

2

1

0

Wang and Evans — Behavior of Iron

167

I 2 3 4 5 6 7 8 910

PARTICULATE Fe(III), mg/liter

6. Relation between particulate re (III) and psiticulate PCh-P.

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Transactions Illinois Academy of Science

Literature Cited

American Public Health Association. 1965. Standard methods for the exami¬ nation of water and wastewater. 12th edition. New York. 769 pp.

Evans, L. T. and E. W. Russell. 1959. The adsorption of humic and fulvic acids by clays. J. Soil Sci. 10:119-132.

Ghosh, M. M., J. T. O’Connor and R. S. Engelbrecht.' 1966. Precipitation of iron in aerated ground water. Proceed¬ ings ASCE, J. Sanitary Eng. Div. 4687 : 199-213.

Hem, J. D. 1960. Survey of ferrous- ferric chemical equilibria redox poten¬ tials. U. S. Geol. Survey Water Supply Paper 1459-B. 55 pp.

Hem, J. D. and W. H. Cropper. 1959. Restraints on dissolved ferrous iron im¬ posed by bicarbonate redox potential and pH. U. S. Geol. Survey Water Sup¬ ply Paper 1459-A. 31 pp.

Larson, T. E. 1967. Chemical oxida¬ tion and reduction of metals and ions in solution, in S. D. Faust ed. Princi¬ ples and applications of water chemis¬ try. John Wiley & Sons. New York. 643 pp.

Lee, G. F. and A. W. Hoadley. 1967. Biological activity in relation to the chem¬ ical equilibrium composition of natural waters, in R. F. Gould ed. Equilibrium concepts in natural water systems. Amer. Chem. Soc., Washington, D. C. 344 pp. Morgon, J. J. and W. .Stumm. 1964. The role of multivalent metal oxides in limnological transformations, as exempli¬ fied by iron and manganese. Proc. Sec¬ ond Internatl. Water Poll. Res. Conf. pp. 103-131.

Shapiro, J. 1964. Effect of yellow or¬ ganic acids on iron and other metals in water. J. Water Works Assoc. 56: 1062- 1082.

Stumm, W. and G. F. Lee. 1961. Oxy¬ genation of ferrous iron. Ind. Eng. Chem. 53:143-146.

Wang, W. C. and D. J. Brabec. 1969. Nature of turbidity in a major river. J. Amer. Water Works Assoc. 61:460- 464.

Wang, W. C. and R. L. Evans.. 1969. Variation of silica and diatoms in a stream. Limnol. Oceanog. 14:941-944.

Manuscript received August 29, 1970

IMPROVED X-RADIOGRAPHY OF CYLINDRICAL

SEDIMENT CORES

GORDON S. FRASER AND ADRIAN F. RICHARDS

Illinois State Geological Survey,

Urbana, Illinois 61801 and

Center for Marine and Environmental Studies,

Lehigh University ,

Bethlehem, Pennsylvania 18015

Abstract. — X-rays passing through the center of cylindrical sediment cores during radiography are absorbed to a greater de¬ gree than are X-rays passing through the sides of the core. Four methods have been devised to compensate for this effect. ( 1 ) The penetrating power of the X-ray beam may be increased. (2) A lead in¬ tensifying screen can be used to increase the intensity of X-rays passing through the center of the core. (3) A material similar to that in the core may be packed around the sides of the core. (4) The film density of the central region of the film may be reduced. These methods were quantita¬ tively compared by testing the effects of each method on the exposure and sensi¬ tivity of the film. The results of the tests indicated that each method alleviated the problem but to a different degree. Method 3 produced the most even exposure across the film and showed the maximum detail of the internal features of the core.

When radiographs of cylindrical sediment cores are made, X-rays passing through the center of the core tube are absorbed by the sedi¬ ment to a greater degree than X-rays passing through the sides of the core. The radiograph is unevenly exposed and the gradient of film density (darkness of film) increases from the center toward the sides of a radio¬ graph. A film correctly exposed for the center of the core will be over¬ exposed near the edges, resulting in a loss of detail.

We tested several methods to mini¬ mize this problem. Only a tentative solution has been reached because each method introduces factors that adversely affect the film sensitivity or the ability of the film to record de¬ tail.

Methods

In each of the methods discussed below, a Picker Gemini Model 160 constant potential industrial radio- graphic machine was used with a 150k V Morris Be-window X-ray tube selected to have a 0.5 mm diameter effective focal spot. The X-ray out¬ put from the tube was collimated by a medical-type collimator attached to the tube. The focal-spot to speci¬ men distance was fixed at 100 cm. Kodak AA industrial X-ray film was used exclusively. The film was de¬ veloped in a Calumet Model 147 ni¬ trogen-burst machine to insure that each film was uniformly processed.

The penetrating power of X-rays may be increased by increasing the potential, or kilovoltage, between the anode and the cathode in the X-ray tube. The “harder” X-rays that are produced at higher kilovoltages are less sensitive to varying thick-

1169]

Transactions Illinois Academy of Science

170

nesses of a cylindrical core and, therefore, reduce the density gradi¬ ent across the radiograph. Radio¬ graphs made from these hard X-rays show less detail of the core because harder X-r ays are also less sensitive to differences of density in the object being radiographed.

A second method uses lead inten¬ sifying screens to increase the inten¬ sity of X-rays passing through the center of the core relative to those passing through the sides. The rays that have passed through the central region are “hard” X-rays of short wave length because the softer rays have been selectively absorbed. To¬ ward the sides of the core, however, more soft rays can penetrate the core because the material is thinner. The intensifying effect increases as wave length decreases (Clark, 1955), so that rays of short wave length penetrating the central region of the core tube are preferentially strength¬ ened. Unfortunately, the ability of the intensifying screens to reduce the film density gradient is somewhat limited.

In the third method, material is packed around the core tube present¬ ing plane parallel surfaces to the incoming X-rays. Thus, X-rays passing through the sides of the core are absorbed to an equal degree as those passing through the center.

Several materials have been used as th$ absorbing material. Klinge- biel et al (1967) immersed the cores in liquids of varying densities. The densities of the liquids were matched with the density of the material of the core so that the absorptive char¬ acteristics of both materials were the same. The liquids, however, were in¬ convenient and difficult to handle

during preparation of the core for radiography.

Haase (1967) attacked this prob¬ lem by placing the cores in molds of plaster or plastic. Plastic was pre¬ ferred because the plaster contained air bubbles and other imperfections. Baker and Friedman (1970) placed cores in a machined aluminum block. The molds are convenient to handle, but their densities cannot be varied to match the absorptive characteris¬ tics of the sediment core. They also may be expensive or difficult to construct.

Bouma (1969) recommended plac¬ ing the core in loosely packed fine sand. The absorptive effect of the sand could be varied by changing the thickness of the sand pack. A fair match between the absorptive characteristics of the sediment and the sand could thus be obtained. Even though a fine sand is used, how¬ ever, the graininess of the sand would be shown on the film and would tend to mask detail in fine¬ grained sediment.

A method which alleviates these problems was devised by Nathan Ayer (1970) in 1967. He designed a plastic box (Fig. 1) containing small glass spheres, called glas-shot® or microshot, as the absorbing ma¬ terial. The box consists of an outer plexiglas shell fitted with an inner

Figure 1. Glass bead box designed by Nathan Ayer. The core tube slides in¬ side the plexiglas tube.

Fraser and Richards— X -radiography of Sediment Cores

171

plexiglas tube that has an inside diameter only slightly larger than the 4.5-inch (11.4-cm) outside diame¬ ter of our core tubes. The microshot ranges from 10 to 53 microns in di¬ ameter. (Number MS-XL, Micro¬ beads Division, Cataphate Corp., Jackson, Miss.) The box is con¬ venient because the core can easily be fitted into the plexiglas tube with¬ out having to repack the microshot each time a new core is radiographed. The microshot is sufficiently small to prevent masking detail in fine¬ grained sediments, and the absorp¬ tive characteristics of the glass beads are similar to those of many types of unconsolidated sediments. In ad¬ dition, the absorptive ability can be varied bv increasing or decreasing the packing density of the microshot. The glass beads, however, cause some scattering of the X-ray beam that tends to blur the image slightly on the radiograph.

In method 4, the film density of the entire radiograph is decreased until detail can be seen along the edges of the film. This procedure, however, may cause the film density of the central region to become too low, causing the visibility of detail in that area of the radiograph to be reduced.

Comparison of Methods

The relative pros and cons of the four methods may be quantitatively compared by testing the effect of these methods on the density gradi¬ ent and on the contrast of the film. A synthetic core was constructed for the tests by putting five strips of lead foil on half the width of a plexi¬ glas plate and inserting the plate into

a plastic core tube filled with the same microshot as those in Ayer’s box (Fig. 2). A radiograph of the

core showed the strips of lead as slightly lighter images against a darker background on one side of the radiograph and a gradient of film density progressively increasing from the center to the edge of the other side of the radiograph (fig. 3).

The film density of the images of the five lead strips and of the corre¬ sponding areas on the opposite side

Figure 3. Typical test radiograph print. The darker areas represent the lead strips and areas enclosed by the dashed lines are the counterpart areas to the images of the lead strips.

172

Transactions Illinois Academy of Science

of the film were measured. To com¬ pute the film contrast, the arithmetic difference was determined between the film density of the image of a lead strip and the film density of its counterpart area on the opposite side of the film. The film contrast also had a gradient from the center to the edge of the radiograph.

Three sets of graphs were prepared to show the effects of the various methods on the density gradient and the film contrast. The first set (Fig. 4) compares the film density gradient for each of the first three methods and for three different kilovoltages. The graphs are plotted with film den¬

sity as a function of distance from the center line of the radiograph of the core tube. The initial film den¬ sity (film density of the center line of the radiograph) was 2.00 in all cases. The angle of the slope of any of the plotted curves bears an inverse relation to the film density gradient.

The second set of graphs (Fig. 5) are plots of the film contrast gradi¬ ent for the first three methods and for three different kilovoltages. The contrast is plotted as a function of distance from the center line of the radiograph with an initial film den¬ sity of 2.00. The angle of slope of

Figure 4. Plot of the film density as a function of distance from the center line of the radiograph for methods 1, 2, and 3 with three different kilovoltages. No film densities could be measured beyond 40 mm from the center line of the film for methods 1 and 2 owing to extreme blackening of the film.

Distance from Center-line of Radiogroph (mm)

Fraser ancl Richards — -X -radiography of Sediment Cores

173

any of the curves bears an inverse relation to the film contrast gradient.

The third set of graphs (Fig. 6) was prepared from data taken from radiographs in which the initial film density was set at 1.00, 1.50, and 2.00, while the kilovoltage was held constant at 70 kV. These graphs were prepared to show the effects of method four (decreasing the ini¬ tial film density) on the density gradient and the contrast gradient.

Figures 4 and 5 show that the microshot box, method 3, is the best mechanism for reducing the film density gradient. This method, how¬ ever, also has the most detrimental effect on the film contrast. In all

graphs, the microshot box method showed the lowest contrast at the center line of the radiograph and also the slowest rate of increase from the center to the edge of the radio¬ graph. Use of the lead intensifying screen in method 2 decreased the film density gradient slightly, but it was not as harmful to the contrast as method 3. Increasing the kilo- voltage decreased the density gradi¬ ent considerably but also adversely affected the contrast. The film den¬ sities at the edges of the radiographs made using methods 1 and 2 were too high to be measured, i.e., no light penetrated these areas when a high- intensity X-ray illuminator was used.

line of the film for methods 1, 2, and 3 using three different kilovoltages. No film densities could be measured beyond 40 mm from the center line for methods 1 and 2 because of extreme blackening of the film.

174

Transactions Illinois Academy of Science

Figure 6 shows that with an initial film density of 1.00 the density gradi¬ ent was the lowest and the contrast of the center portion of the radio¬ graph was so low that the contrast gradient became negative. Increas¬ ing1 the film density to 1.50 increased the contrast but also greatly in¬ creased the film density gradient. An initial film density of 2.00 showed the best contrast and also the most extreme film density. In all tests, the edges of the radiograph were completely blackened, and it was im¬ possible to measure the film density with a high-intensity illuminator.

Although the evidence of the graphs seems inconclusive, it should

be pointed out that an extreme film density gradient is much more ad¬ verse to radiographic sensitivity than a steep contrast gradient is beneficial. A satisfactory compromise is found by decreasing the density gradient and accepting the slightly adverse effects produced on the contrast of the film.

Methods 1, 2, and 4 left 10 to 15 mm of a radiograph completely opaque when viewed on the available illuminator. Method 3 was the only method tested that produced accept¬ able film densities across the entire radiograph and showed the maximum detail of internal features on X-radi- ographs (Fig. 7).

Figure 6. Plot of the film density and the contrast as a function of the distance from the center line of the film for three initial film densities (IFD), 1.00, 1.50 and 2.00. Blackening of the film beyond 40 mm from the center line produced film den¬ sities too high to be measured.

Fraser and Richards — X -radiography of Sediment Cores

175

Discussion

A film density of about two is considered optimum in industrial ra¬ diography. Lower film densities lack detail, and higher densities are diffi¬ cult to view with conventional high- intensity viewers. A film density of two is opaque to the eye when viewed without the use of a high-intensity illuminator. AYhile optimum detail of the film negative is possible using the viewer, we have found that it is difficult to obtain satisfactory pos¬ itive prints. The dynamic range of most photographic papers evidently is insufficient to satisfactorily record the detail present in the negative. Very long exposure times also were necessary to obtain suitable prints from the dense negatives. Small dif¬ ferences in film density caused by the geometry of the radiographic box used in method 3, in particular, re¬ sulted in unsatisfactory prints made by conventional photographic pro¬

cedures. A number of methods were tried to solve this problem. The most satisfactory results were ob¬ tained using the Mark IT Log-Etron- ic contact printer.

Acknowledgments

The studies reported were made using the X-ray facility of the Sedimentology Laboratories in the Department of Geology, LIniversity of Illinois, Urhana. The X-ray machine was acquired from funds pro¬ vided by NSF Grant GK-1292 to A. F. Richards and J. E. Stallmeyer. Our work at the University of Illinois was supported by Office of Naval Research Contract NONR 3985 (09), NR 081-260. The manuscript was partly prepared under Of¬ fice of Naval Research Contract N00014- 67-A-0370-0005, NR 083-248. The posi¬ tive print shown in figure 7 was kindly made for us at the Waterways Experiment Station, Vicksburg, Mississippi, through the courtesy of E. L. Krinitzsky.

References

Ayer. Nathan. 1970. Radiography of sediment cores from southern Lake Mich¬ igan: Ill. State Geol. Surv. Environ¬ mental Geol. Note, in preparation.

lii ' Ji

Figure 7. X-radiograph print using the microbead box of a sediment core within a 1 1 -cm-diameter core barrel of Delrin plastic. A gastropod shell is the dominent feature in the core. Smaller features probably are clam shells.

176

Transactions Illinois Academy of Science

Baker, S. R. and G. M. Friedman. 1970. A non-destructive core analysis technique using X-rays: Jour. Sed. Petrol., 39, p. 1371-1383.

Bouma, A. H. 1969. Methods for the study of sedimentary structures: John Wiley and Sons, New York, 458 pp. Clark, G. L. 1955. Applied X-rays: McGraw Hill Book Company, Inc., New York, 843 pp.

Haase, M. C. 1967. X-radiography of unopened soil cores: U. S. Army Eng. Waterways Experiment Station Misc. Paper No. 3-918, 24 pp.

Klingebiel, Andre, Alain Rechiniac, and Michel Vigneaux. 1967. Etude ra- diographique de la structure des sedi¬ ments meubles: Marine Geology, 5, p. 71- 76.

Manuscript received June 15, 1970.

THE SPATIAL DISTRIBUTION OF LAKE-EFFECT SNOWFALL WITHIN THE VICINITY OF LAKE MICHIGAN

KENNETH FREDERIC DEWEY

Department of Geography,

University of Toronto, Toronto, Ontario, Canada

Abstract. — The average monthly and annual snowfall patterns are analyzed for the region within 100-150 miles of Lake Michigan. From these patterns the dis¬ tribution of lake-effect snowfall is deter¬ mined for the study region.

During the last few years much attention has been given to Great Lakes Climatology. Among the many parameters being studied is snowfall. It has been illustrated by several authors that excessive amounts of snowfall occur along the shorelines of the Great Lakes, especially along the lee shore of the lakes (Falconer, Lansing, and Sykes, 1964 ; Sheridan, 1941 ; Pack, 1963 ; Namias, 1960 ; Bolsenga, 1967 ; Johnson and Mook, 1953; and Williams, 1963). Muller (1966, p. 256) developed a map for mean seasonal snowfall in the Great Lakes region and surrounding areas. The most outstanding features of this map are the snowbelts associated with the frequently recurring lake squalls coming off the Great Lakes.

These excessive amounts of snow¬ fall along the shores of the lakes have been explained to be an overt manifestation of lake-effect snow showers. Lake-effect snowfall by defi¬ nition occurs as a result of cold air being advected over the warm moist surface of a lake. The air in contact with the surface warms rapidly, gains moisture and under superadi-

abatic lapse rate conditions rises rap¬ idly forming convective clouds and precipitation. During the periods of lake-effect snowfall it is assumed that the snowfall is generated specifically from the vast reservoir of heat and available moisture in the lake, and that there is no direct influence from either cyclonic or frontal systems.

Purpose of the Study

It is the purpose of this study to conduct an analysis of the phe¬ nomenon lake-effect snowfall for a single lake in the Great Lakes basin. Lake-effect snowfall is studied in¬ directly by analyzing the spatial variation of snowfall within the vicinity of Lake Michigan. Aver¬ age monthly and annual snowfall amounts are determined for the study region and the patterns an- al}rzed.

The Study Area

A boundary was chosen for this study to average between 100 and 150 miles inland from the shoreline of Lake Michigan. Lake Michigan offers several advantages which do not exist for any of the other Great Lakes. There is a dense network of reporting stations along all sides of the lake. There is topographic uni-

[177]

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Transactions Illinois Academy of Science

formity throughout most of the study region. And, there is little interac¬ tion between Lake Michigan and the other lakes in the Great Lakes basin.

Sources and Utilization of Data

The U. S. Department of Com¬ merce’s data records for each of the states within the region of study were examined. A list was compiled indicating all reporting stations in the study area that kept systematic monthly and annual records of snow¬ fall. A total of 145 stations were in¬ cluded in the study.

Monthly and annual snowfall amounts were tabulated for the sta¬ tions in the study region for 10 successive snowfall seasons begin¬ ning October 1959 and ending March 1969. Monthly snowfall amounts were tabulated for the four months November to February of each snow season. An average monthly and an¬ nual snowfall amount was computed for each of the 145 stations in the

study.

•/

These averages were compared to the long-term averages which were available in the “ Climatography of The United States” series. It was noted that the 10 year averages ob¬ tained in this study were significant¬ ly higher than the long-term aver¬ ages. The average increase over the study region was from 10% to 15%. In his recent study, Eichenlaub (1970) found that there has been a 100% increase in the average snow¬ fall amounts of western Michigan. There is no definite answer to the question of what is causing this in¬ crease in snowfall and lake-effect snowfall. There are however several

possible explanations for this climat¬ ic change which warrant further research. Namias (1960) suggested that an increase in snowfall can re¬ sult from a shift in the position of the mean troughs and ridges over North America. It has also been suggested that a general cooling of the atmosphere has been occurring since the 1930 ’s. Finally further research should investigate the role of atmospheric pollution in increas¬ ing the amount of snowfall (especial¬ ly near the large industrial com¬ plexes like Chicago-Gary, Milwaukee, and Muskegon).

Analysis of The Average Annual Snowfall Pattern

The average annual snowfall was portrayed on the map of the study region using an isopleth interval of 10 inches (Figure 1). In an analysis of the annual as well as the monthly snowfall patterns it should be em¬ phasized that the positioning of the isopleths across the lake is based upon estimation. Although there are values of snowfall at the shoreline of each side of the lake it is difficult to determine the correct gradient of isopleths across the water. Accord¬ ing to Changnon (1968, p. 23) “When lake-effect snowfall develops over the lake it begins somewhere within 20 miles of the eastern shore¬ line.” On the annual snowfall map and all maps of monthly average snowfall, it was found in this study that the isopleths generally tended to parallel both the eastern and west¬ ern shorelines, so it seemed most like¬ ly that the isopleths over the lake would also tend to follow a north-

Dewey — Lake-Effect Snowfall

179

Figure 1. Average Annual Snowfall In Inches.

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Transactions Illinois Academy of Science

south direction. However, it was also assumed that the isopleths of snow¬ fall are more tightly spaced along the eastern shore than the western shore.

Were there no Lake Michigan one would expect the isopleths to be oriented in an East- West direction across the study region, with snow¬ fall increasing to the north. How¬ ever, this latitudinal positioning of isopleths occurs only within 75 miles of the western edge of the study re¬ gion, within 50 miles of the southern boundary of the study region, and at a maximum, within 50 miles of the eastern edge of the study region. The remainder of the study region has a snowfall pattern which could not exist without the presence of Lake Michigan.

Along the western side of the lake, the gradient of snowfall is small. For example, the distance between the 30 inch and 40 inch isopleth is 200 miles. The gradient becomes steeper near the northern margin of the study region, but this is a result of the additive influence of lake-ef¬ fect snowfall from Lake Superior. The extent of lake-effect snowfall averages 10-15 miles inland along the western shore and the magnitude averages 10-15 inches annually. Lake- effect snowfall therefore increases the annual snowfall along this side of the lake by 30-40%.

The additive factor of lake-effect snowfall is dramatically illustrated along the lee side of the lake. Here the isopleths are packed quite closely together, their high gradient indicat¬ ing a region of copious snowfall. The penetration inland of lake-effect snowfall averages 50-75 miles in

Michigan. Some areas receive more than 60 inches of lake-effect snow¬ fall annually, which is 200% more snowfall than at the same latitude in the interior of the state.

There are three significant fea¬ tures in the pattern of isopleths along the eastern shoreline of Lake Michigan. First, there are three cores of extremely heavy snowfall each averaging over 130 inches. The first two cores, centered on Gaylord-Van- derbilt and Maple City in northern Lower Michigan are a function of elevated topography, as well as a peninsular effect. The land area of this region is surrounded on three sides by the close proximity of water bodies therefore increasing the mag¬ nitude of lake-effect snowfall. Ele¬ vations above sea level for these two cores respectively are approximately 1435 feet and 850 feet, or 853 feet and 268 feet respectively above the mean level of Lake Michigan (582 feet) .

The effect of these hills is twofold, one effect is to provide lifting, and the other is to provide increased sur¬ face friction for the bands of snow as they move inland from the lake. In an article by R. L. Peace and R. B. Sykes (1966) which examined the conditions attendant upon lake- effect storms at the eastern end of Lake Ontario, it was made evident that the lake-effect snow bands are characteristically shallow systems with radar-detectable tops usually lying below 10,000 feet. Extreme in¬ stability is also attendant with these bands. While the relief of these two areas in northern Lower Michigan may be insignificant for orographic snowfall during cyclonic and frontal

Dewey — Lake-Effect Snowfall

181

situations, it is significant for oro¬ graphic snowfall during lake-effect snowfall situations.

The third core centered on Mus¬ kegon could be related to topography but not with as much confidence. Instead, the primary factors causing excessive snowfall over the Muskegon area are hypothesized to be pollution and urban influence. Several recent studies have indicated that there has been a considerable increase in the amount of air pollution downwind from Great Lakes metropolitan cen¬ ters. The presence of high concen¬ trations of ice nuclei and crystals in the Great Lakes area and the avail¬ ability of moist air from off the lake is favorable for the occurrence of increased lake-effect snowfall. No sig¬ nificant evidence is available at the present time to confirm such a link. Therefore, with pollution becoming a more urgent problem there is the need for further research into the role of pollution in lake-effect snow¬ fall. The urban effect results from heat being added to the bands of snowfall as they move inland from the shoreline. The heat added to the bands of snowfall increases the in¬ stability of the air masses and sub¬ sequently the fall of snow for the area surrounding Muskegon.

The hills of western Michigan in¬ crease the surface friction greatly as these bands of snowfall move across the region and this creates the second significant feature of the pattern of isopleths. This increased surface friction for areas of hills, and conversely, the relative lack of surface friction for areas of little local relief, creates a phenomenon which can be termed ‘ ‘ avenue of pen¬ etration’ \ It is hypothesized that

where there are hills close to the shore of the lake, large amounts of snowfall result from the fact that the increased surface friction slows the storm bands down so that much of the energy is released near the shore¬ line and little residual energy or moisture remains by the time the band has crossed the hilly region. Conversely, where there are no topo¬ graphic barriers along the shore, the bands of snowfall can progress in¬ land with only a gradual loss of moisture and most likely extend fur¬ ther inland creating avenues of pene¬ tration.

The third significant feature in the pattern of isopleths as illustrat¬ ed on Figure 1 is the occurrence of the maximum amounts of snowfall inland away from the shoreline. The maximum amount of lake-effect snowfall is felt at the shoreline of the western side of the lake with decreasing amounts of snowfall oc¬ curring inland. However, in Michi¬ gan (not including the Upper Pen¬ insula) the maximum amounts of snowfall occur approximately 25 miles inland. This is in agreement with the findings in Changnon’s (1968a) study of annual snowfall in the Lake Michigan basin. In his study, Changnon found that the maximization of lake-effect snowfall occurs anywhere from 10 to 25 miles inland, with 10 to 40 more inches of snow there annually than at the immediate eastern shoreline of Lake Michigan.

It is hypothesized that this phe¬ nomenon exists as a result of the increased surface friction as the air passes from the lake onto the land surface of Michigan and Indiana, causing the air masses to slow down

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Transactions Illinois Academy of Science

and begin to pile up inland some dis¬ tance from the shoreline. The in¬ creased friction resulting from rough topography can enhance this process of convergence of air masses as the bands of lake-effect snowfall move inland.

Comparison of The Average Monthly Snowfall Patterns with The Annual Pattern

The average monthly snowfall pat¬ terns for each of the four months under investigation were portrayed on Figures 2-5 using an isopleth interval of 5 inches. As in the case of the annual average snowfall, the positioning of the isopleths across the lake was based upon estimation.

The three cores of extremely heavy snowfall which appeared on the map of annual snowfall appear only from December through February. Dur¬ ing November the second core of heavy snowfall around the vicinity of Maple City has only begun to de¬ velop. The third core of heavy snowfall centered over Muskegon is well developed by this time. There is the development of a fourth core of snowfall south of Muskegon for all of the months under study except January. The significant feature of this core of snowfall is that the di¬ ameter is considerably larger, aver¬ aging three times the size of the largest of the other three cores. Dur¬ ing January the isopleths of snow¬ fall around the core of snowfall cen¬ tered on Musekgon curve southward and are elongated to such a degree that the area represented by the fourth core might be considered to be a part of the Muskegon core.

The distributions of the average

monthly snowfall also dramatically display the avenues of penetration. For all months under investigation, the tendency for fingers of heavier snowfall to extend inland is quite definite. The eastward extent of the avenues of penetration remains al¬ most identical throughout the re¬ mainder of the snow season. The only variance from month to month is in the intensity of snowfall or amount of snowfall represented by the avenues of penetration.

As in the distribution of the aver¬ age annual snowfall, there is the ten¬ dency for the heaviest amounts of snowfall to occur inland some 20 to 30 miles from the eastern shoreline for each of the months under inves¬ tigation. It is significant to note that whether the month under in¬ vestigation averages little snowfall or copious amounts of snowfall, the heaviest snowfall area remains iden¬ tical.

Conclusions

An analysis of the annual and monthly snowfall patterns within the vicinity of Lake Michigan revealed an uneven spatial distribution of snowfall. This uneven distribution of snowfall results from the additive influence of lake-effect snowfall. The average distribution and magnitude of lake-effect snowfall was deter¬ mined from these patterns. There were three significant patterns of lake-effect snowfall which appeared on all the maps of snowfall : Cores of excessive snowfall ; avenues of penetration ; and, a tendency for the heaviest amounts of snowfall to oc¬ cur inland 20-30 miles from the shoreline.

Dewey — Lake-Effect Snowfall

183

Figure 2. Average November Snowfall Pattern (5 inch interval).

184

Transactions Illinois Academy of Science

Figure 3. Average December Snowfall Pattern (5 inch interval).

Dewey — Lake-Effect Snowfall

185

Figure 4. Average January Snowfall Pattern (5 inch interval).

186

Transactions Illinois Academy of Science

Figure 5. Average February Snowfall Pattern (5 inch interval).

Dewey — Lake-Effect Snowfall

187

Literature Cited

Bolsenga, S. J. 1967. Great Lakes snow depth probability charts and tables. U. S. Lake Survey Research Report No. 5-2. 7° pp.

Changnon, S. A. 1968. Precipitation climatology of Lake Michigan basin. Illinois State Water Survey Bulletin 52. 46 pp.

Eichenlaub, V. L. 1970. Lake effect snowfall to the lee of the Great Lakes: its role in Michigan. Bulletin AMS. 51: 403-412.

Falconer, R. L., L. Lansing, and R. Sykes. 1964. Studies of weather phe¬ nomena to the lee of the eastern Great Lakes. Weatherwise. 17: 256-261. Johnson, E. C., and C. P. Mook. 1953. The heavy snowfall of January 28-30, 1953, at the eastern end of Lake Ontario. Monthly Weather Review. 81: 26-30.

Muller, R. 1966. Snowbelts of the Great Lakes. Weatherwise. 19: 248-257. Namias, J. 1960. Snowfall over the east- tern United States: factors leading to its monthly and seasonal variation. Weatherwise. 13: 238-247.

Pack, A. B. 1963. The heavy snows at Watertown, New York. Weatherwise. 16: 66-67, 78.

Peace, R. L., and R. B. Sykes. 1966. Mesoscale study of a lake-effect snow¬ storm. Monthly Weather Review. 94: 495-507.

Sheridan, L. W. 1941. The influence of Lake Erie on local snows in western New York. Bulletin AMS. 22:393-395. Williams, G. C. 1963. An occurrence of lake-snow — one of the direct effects of Lake Michigan on the climate of the Chicago area. Monthly Weather Review. 91: 465-467.

Manuscript received July 29, 1970

THE COUPLING OF ENERGY PRODUCTION TO SYNTHESIS IN THE ORIGINAL OPERATION OF LIVING SYSTEMS

NEWTON RESSLER

Departments of Pathology and Biochemistry, University of Illinois Medical Center, Chicago, Illinois

Abstract. — The development of the self duplication of macromolecules into a living system is usually considered either to be due to heat energy, or without any explicit regard for the required energy supply. Sim¬ plified thermodynamic models are presented to illustrate that kinetic energy alone may not have been able to provide the neces¬ sary characteristics for the development of the synthesis of macromolecules into life processes. Reasons are presented for be¬ lieving that the synthesis could only have developed in living systems after coupled to exergonic reactions.

Investigators concerned with the various phases of the origin of life processes, such as primitive photo¬ synthetic reactions or the develop¬ ment of replication, have not gener¬ ally related them to a coupling of endergonic with exergonic reactions. It is quite common to discuss the development of self duplication of macromolecules into primitive bio¬ logical systems without explicit re¬ gard for the energy supply neces¬ sary for the process (Gaffron, 1965). In this communication, I shall con¬ sider some reasons why the coupling of energy producing reactions to syn¬ thesis or reproduction may have been a primary prerequisite for the emer¬ gence of original life processes. Jus¬ tifications for the explicit inclusion of this coupling in the definition of a living system will be examined.

This problem might be approached

by attempting to derive the most simple systems which still retain cer¬ tain characteristics which are essen¬ tial for present forms of life. A description of primary, essential characteristics (and their distinction from secondary ones) is difficult to make in an objective or unarbitrary manner. Nevertheless, the following three features may appear sufficient¬ ly reasonable for use as a tentative hypothesis for the present purpose :

1. The direct or indirect collec¬ tion and storage of solar en¬ ergy. Photosynthesis could be regarded as a direct collection, while the consumption by non- photosynthesizing or lieterotro- phic organisms of photosyn¬ thetic products would then constitute an indirect collec¬ tion. Insofar as the products of photosynthesis have a high¬ er potential energy than the reactants, they represent a form of stored energy which can later be released by exer¬ gonic reactions.

2. The coupling or controlled use

of this energy for synthesis of additional energy collecting structures : i.e., reproduction

or growth.

3. The necessity of a specific mo-

[188]

Ressler — Energy Coupling

189

lecular geometry or structure for the above processes. A specifically ordered molecular geometry (as in chloroplasts, mitochondria, etc.) is general¬ ly recognized as a universal characteristic of present forms of life (Dean & Hinshelwood, 1966).

Attempts to derive the most simple system which still retains these three characteristics might lead to ther¬ modynamic models (without specify¬ ing reactants) such as those in Fig¬ ure 1. In Figure la, for example, conformation I consists of one or more substances whose molecules are

Potential

energy

Coupled | reaction*- -/

6

t

Conformation

solar energy

A - Conformation

i

Synthesis of X

C - * - Constituents

a

I'

I

of I

Potential

energy

Substance H

Constituents

of n

Solar

energy

♦ . . Coupled . reactions

♦

Synthesis of

Conformation I

I

Constituents of I

b

Figure 1. a. Energy for coupling stored by a photo-induced high energy conformation. The potential energy level A is that of a structure whose molecules are in a particular geometrical arrange¬ ment necessary for acceptance and storage of light energy. This is done by a light energy induced change from conformation I to a conformation of higher potential energy, T, (at potential energy level B). Upon reverting back to conformation I, the energy liberated is used for autocatalytc synthesis of more material of structure I, from constituents of potential energy C.

b. Energy for coupling stored by the photo-induced formation of intermediate substance II, of higher potential energy than that of the reactants. The energy liberated by the exergonic reactions (or metabolism) of II is coupled to the syn¬ thesis of additional material of structure I.

in a particular geometrical arrange¬ ment essential for the utilization of solar radiation. When such radia¬ tion occurs, conformation I stores this energy by changing to a differ¬ ent conformation of higher potential energy, V. When the conforma¬ tion returns from I' to I, the energy liberated is used for (or coupled to) the synthesis of more of the sub¬ stance in conformation I (from con¬ stituents in the surroundings). This synthesis requires the initial presence of the structure being syn¬ thesized, which acts both as an auto- catalyst for its self reproduction and as an acceptor and storer of light energy for this purpose.

In analogy with some concepts of present forms of photosynthesis, the conformational change from I to V could be associated with other con¬ formation — dependent processes, such as oxidation-reduction reac¬ tions, or cation-proton exchanges. A photooxidation-reduction reaction, for example, could depend upon a conformational change if the change brings the bound groups which be¬ come oxidized and reduced close to¬ gether.

Figure lb illustrates a slightly more complex model. The solar ener¬ gy is stored by the light-induced, en- dergonic formation of a separate compound, substance II, such as car¬ bohydrate or ATP. Substance II is thus a form of stored energy, since it has a greater potential energy than the reactants. The reverse re¬ action, or metabolism of substance II, liberates the energy, which is coupled to the synthesis of the ma¬ terial (in conformation I) with the specific structure necessary to cata¬ lyze both the photoreaction and its

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Transactions Illinois Academy of Science

self reproduction.

These models retain the required characteristics in simplified or primi¬ tive systems. Although not essen¬ tial to the main argument of this paper, the energy source has been represented as solar radiations, uti¬ lized via endergonic photoreactions. Daylight has been the only continu¬ ous source of useful energy, and is commonly considered the most prob¬ able energy source for original life s}\stems (Gaffron, 1962).

A specific molecular geometry or arrangement may be essential for various reasons. It is difficult to dem¬ onstrate endergonic photoreactions with a high quantum yield in the laboratory. Gaffron (1965) has sug¬ gested that living organisms are uniquely able to obtain high quan¬ tum yields in such reactions because of the specific structural arrange¬ ment of the molecules. These mo¬ lecular arrangements may serve to prevent the reversal or rapid re¬ combination of reaction products, which would occur in a homogenous solution. Such a molecular arrange¬ ment can be considered metastable, due to its relatively high potential energy.

The models, or others that might be proposed, are meant only to illus¬ trate how the coupling of exergonic reactions to the energy requiring synthetic reactions can be essential for the most primitive systems which still retain the components of a rea¬ sonable definition of life. Let us consider how this coupling can lead to an energy source which is utilized in a controlled manner ; i.e., so that the extent of the energy collection and storage, energy liberation, and endergonic synthetic reactions are

each correlated with the others ac¬ cording to the overall requirements of the system. If the molecules of the system are all in conformation I' in the case of Figure la, or if all of the binding sites for substance II are occupied in the case of Figure lb, then further energy cannot be stored beyond this maximum value, until some of the stored energy is utilized. The stored energy can only be used when the availability of the neces¬ sary constituents in the medium per¬ mits the energy-requiring synthesis to take place. The coupled use of the stored energy for this purpose would result in the reformation of conformation I from I' in the case of Figure la, or the vacating of some of the binding sites for substance II in the case of Figure lb. Further en¬ ergy storing photoreactions can now take place. The coupling thus serves to limit the energy storage and liber¬ ation according to the requirements, which are determined by the extent of synthesis. Without the coupling, the energy uptake and liberation might proceed independently at max¬ imum rates at a time when no sub¬ strates necessary for synthesis are available. In this case, the corre¬ lation of activities which is observed in present forms of life would not exist.

The mechanism of such coupling, i.e., the means of energy transfer from the site of energy liberation to the site of utilization, is also a primary consideration. When pho¬ tosynthesis occurs in presently exist¬ ing organisms, there is evidence that the radiant energy is transferred to, and used at, the reaction centers by means of a resonant transfer mech¬ anism (Rabinowitch, 1963). Reso-

Resslcr — Energy Coupling

191

nant energy transfers are also a pos¬ sible mechanism in the coupling of energy liberating reactions with syn¬ thetic ones. Various characteristics of resonant energy states make this possibility of interest (Rabinowitch, 1963; Ressler, 1969). The efficiency of energy transfer by a resonant mechanism depends upon the mole¬ cular geometry, which is consistent with the necessity for a specific molecular geometry or structure in living systems. The efficiency also de¬ pends upon the overlapping of the energy levels of the energy donor and acceptor. This provides the op¬ portunity for energy “ switches”, or activation of reactions in a specific sequence. Resonant energy transfers can occur at both electronic and in¬ fra-red frequencies, so that steps with various energy requirements might be activated. Since this type of transfer does not require molecular contact or heat, it can occur with little increase in entropy. The pos¬ sibility of this mechanism playing a role in biological processes such as enzyme activation and the control and coordination of energy trans¬ duction, has recently been discussed by the present author (1969).

The coupling of energy liberating and energy requiring reactions could, of course, also involve other factors. In Figure lb, for example, the me¬ tabolism of substance II might be re¬ quired because it liberates one of the substrates involved in the syn¬ thesis of the substance in conforma¬ tion I. Whatever mechanisms may be involved, the energy requiring synthetic reactions must be coupled to an energy source in order for reproduction to occur. Without a regular source of energy, the repro¬

duction of macromolecules would only be a temporary or transient phenomenon, without permanent con¬ sequences.

It might be assumed that the en¬ ergy for initial life systems could have been provided by heat, without coupled, energy producing reactions. Even though a certain amount of kinetic energy, i.e., a given tempera¬ ture range, may be necessary, this explanation still has certain limita¬ tions. Hulett (1969) has discussed reasons why the prebiological syn¬ thesis of intermediate substances, later used for development towards living systems, probably required a coupled energy source, rather than heat. The same considerations would be relevant when such substances started to become synthesized into biological systems. Biological struc¬ tures involve a high degree of order, and a relatively high potential en¬ ergy state. Heat or kinetic energy would increase the entropy or dis¬ order. The stability of such a me¬ tastable state, and the accuracy of the reproduction would therefore tend to decrease at higher tempera¬ tures. The requirement of a specific molecular geometry for heat acti¬ vated reactions is not apparent. It would also be difficult to account for the correlation and adaptation of different activities with changes in the environment, if heat were the sole energy source.

Theories concerned with the de¬ velopment of the synethesis of macro¬ molecules into primitive life systems, which do not explicitly consider the energy source required, generally in¬ volve an implicit assumption that the reactions occur by virtue of heat energy. The large majority of such

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Transactions Illinois Academy of Science

theories do not specifically include energy-producing reactions coupled to this synthesis. Since this coupling would appear to be necessary before such synthetic reactions could devel¬ op into biologically functioning sys¬ tems, it is hoped that this communi¬ cation may be of value.

Acknowledgments

Dean, A. C. R. and Hinshelwood, Sir C. 1966. Growth, Function and Regulation in Bacterial Cells, London, Oxford Press.

Gaffron, H. 1965. In The Origins of Pre- Biological Systems, S. W. Fox, ed., New York, Academic Press, p. 437

- . 1962. In Horizons in Bio¬ chemistry, M. Kasha and B. Pullman, eds. New York; Academic Press, p. 59.

H ulett, H. R. 1969. In Limitations in Pre-Biological Synthesis, J. Theoret. Biol., 24, p. 56.

Rabinowitch, E. 1963. In Fifth Interna¬ tional Symp in Biochemistry, H. Timiya, ed., New York, Pergamon Press, p. 14.

Ressler, N. 1969. In Control of Cellu¬ lar Processes by the Coupling of Reso¬ nant Energy to Hydrogen Transfer Re¬ actions, J. Theoret. Biol., 23, 425.

Manuscript received January 26, 1971

NOTES

GIDEON HERMAN BOEWE 1895-1970

ROBERT A. EVERS AND J. CEDRIC CARTER

State Natural History Survey, Urbana, Illinois 61801

G. H. Boewe in 1963

Gideon Herman Boewe, associate plant pathologist, Illinois Natural History Sur¬ vey, and a member of the Illinois State Academy of Science, died suddenly in his home in Champaign, Illinois, on Satur¬ day, December 19, 1970, at the age of 75 years.

Mr. Boewe was born October 3, 1895, in Parkersburg, Richland County, Illinois, a son of Henry M. and Augusta C. Maas Boewe. He spent his boyhood years on a farm and attended grade schools in Rich¬ land County. He graduated from the

Olney Township High School. From 1918 to 1924 he taught in grade schools in Lawrence and Richland Counties. On June 12, 1921, Mr. Boewe married Isabelle Shafer in Olney. For nearly 50 years the Boewes were a devoted couple.

Mr. Boewe attended Illinois State Normal University and Eastern Illinois State Teach¬ ers College (now Eastern Illinois Univer¬ sity), receiving a Bachelor of Education degree from the latter in 1928. He then enrolled in the University of Illinois and obtained a Master of Science degree in 1930. Mr. Boewe joined the staff of the Illinois Natural History Survey as field botanist on March 17, 1930. He became assistant plant pathologist in 1947 and associate plant pathologist in 1955. His work was primarily concerned with the distribution, severity, and incidence of field, forage, fruit, and vegetable crop diseases. He was, however, interested in plant dis¬ eases in general and collected numerous specimens which form a large part of the plant disease survey herbarium of the Survey. From 1933 to 1946, the accumu¬ lation of Illinois vascular plants for the Survey herbarium was added to the duties of the plant pathologists on the staff. Mr. Boewe conscientiously assumed this duty and collected numerous samples in con¬ junction with his work on the plant dis¬ ease survey. Mr. Boewe retired in 1966, after 36 years in plant pathology at the Survey.

In the course of his field work, Mr. Boewe discovered a number of plant dis¬ eases new to Illinois. Among these were the downy mildew of soybean in 1929, a tiny toadstool on crop plants in 1935, char¬ coal rot of potatoes in 1941, and Helmin- thosporium blight of oats in 1947. In 1964, his report on plant diseases new to Illinois during the period of 1922-1964 included 123 organisms that attack 101

[193]

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Transactions Illinois Academy of Science

host plants. Plant pathologists in Illinois, and those in the corn belt, perhaps remem¬ ber him best for his publications on field crop diseases and for his annual forecasts, beginning in 1948, on the late-season or leaf-blight stage of Stewart’s disease of corn.

Mr. Boewe was active in his church, the First United Methodist Church of Champaign. He and his wife were mem¬ bers for 40 years. He not only attended regularly but served as an usher for 25 years, was a Church School teacher for many years, and, during the past 10 years was secretary of the Church School. For many years he served on the Official Board. He was also an active member of the Men’s Club of the church. Those of us who associated with Boewe at work, soon learned that his religion was not a “Sun¬ day cloak” but a vital part of his life which was exemplified daily by his kindness and generosity to his fellow men.

Mr. Boewe was a member of the Illinois State Academy of Science for many years and he attended the annual meetings regu¬ larly. In 1944, he was appointed to the Membership Committee, serving as a mem¬ ber until 1950 when he was appointed chairman. He served as chairman until 1959 when, at the annual meeting and at his request was retired from the position. Under his chairmanship the Academy grew, reaching the highest membership in its his¬ tory. He served on the Council of the Academy from 1960 to 1964, was a mem¬ ber of the Nominating Committee in 1962- 63, and a member of the Resolutions Com¬ mittee for a number of years, beginning in 1963. He truly was devoted to the work of the Academy.

In addition to the Academy, Mr. Boewe was a member of the American Association for the Advancement of Science, the Ameri¬ can Phytopathological Society, the Illinois State Horticultural Society, and the Illi¬ nois Vegetable Growers Association. He was also a member of the men’s division of the W.C.T.U., serving as Treasurer of the local union and the county unit since

1967.

He is survived by his wife, and brother, Ellis, of West Salem, Illinois. He was preceded in death by his parents, one sis¬ ter, and three brothers.

Publications of Gideon Herman Boewe

1930

Brown rot (Clerotinia sp.) on flowering

almond. Plant Disease Reporter 14- (11) : 94.

1935

Soybean downy mildew in Illinois. Ibid. 19(16): 257-258.

1936

Mosses from Apple River Canyon, Missis¬ sippi Palisades and White Pines Forest State Parks (with Stella Holmes Bar- rick and Stella M. Hague). Ill. State Acad. Sci. Trans. 28(2) : 83-84. (Vol¬ ume dated Dec. 1935).

The relation of ear rot prevalence in Illi¬ nois corn fields to ear coverage by husks. Ill. Nat. Hist. Surv. Biol. Notes No. 6, (Mimeo) 19 p.

Peach leaf curl in Illinois. Plant Disease Reporter 20(9) : 147.

The relation of ear rot prevalence in Illi¬ nois cornfields to ear coverage by husks. Ibid. 20(10): 165-172.

1938

Tiny toadstools on crop plants in Illinois. Ill. State Acad. Sci. Trans. 30(2) : 103- 104. (Volume dated Dec. 1937).

Peach leaf curl in Illinois. Plant Disease Reporter 22 ( 1 1 ) : 199-201.

Naucoria on small grains in Illinois. Phyto¬ path. 28(11) : 852-855.

Ergot on barley in Illinois. Plant Disease Reporter 22(14): 287-288.

1939

Diplodia ear rot in Illinois cornfields. Ill. State Acad. Sci. Trans. 31 (2): 92-93. (Volume dated Dec. 1938).

Diseases of small grain crops in Illinois.

Ill. Nat. Hist. Surv. Circ. 35. 130 p.

Range extensions for Naucoria cerealis in Illinois in 1938 and a key to certain species in the genus. Plant Disease Re¬ porter 23(2): 24-27.

Peach leaf curl in Illinois in 1939. Ibid. 23(15): 254-258.

Epidemic of bitter rot of apples in south¬ ern Illinois in 1939. Ibid. 23(17) : 294. Charcoal rot in Illinois (with L. R. Tehon). Ibid. 23(19) : 312-321.

1940

Fruit disease problems in Illinois. Trans.

Ill. State Hort. Soc. 74: 154-159. Crown rust of oats in Illinois. Plant Dis¬ ease Reporter 24(14): 302-303.

Peach leaf curl in Illinois. Ibid. 24(14): 305-307.

Stem rust of oats in Illinois. Ibid. 24(15) : 331.

Alfalfa downy mildew in Illinois. Ibid. 24(15): 332.

Bitter rot of apples in southern Illinois in 1940. Ibid. 24(16): 346.

Notes

195

1941

Peach leaf curl in Illinois in 1941. Ibid. 25(13): 355-358.

1942

Charcoal rot on potatoes in Illinois. Ibid. 26(6): 142-143.

Peony anthracnose found in Illinois (with D. B. Creager). Ibid. 26(12/13): 280- 281.

1943

Scab and glume blotch of wheat in Illinois.

Ibid. 27(12/13): 251.

Cereal smuts and rusts in Illinois (with L. R. Tehon). Ibid. 27(17): 343-347. Survey for corn diseases in Illinois (with R. C. Baines). Ibid. 27(22): 611-613.

1944

Damage by smuts to small grains in Illinois in 1944 (with L. R. Tehon). Ibid. ^ 28(24) : 789-791.

Fruit diseases in western Illinois (with J. S. Tidd). Ibid. 28(32): 991-993. Corn diseases in North Central Illinois (with J. S. Tidd). Ibid. 28(35) : 1077- 1079.

1945

Violet scab found in Illinois for the first time (with J. L. Forsberg). Ibid. 29- (25/26): 680.

1946

Late blight in southern Illinois. Plant Disease Reporter Suppl. 164: 293-294. New oats disease is widespread in Illinois in 1946 (with L. R. Tehon). Plant Disease Reporter 30(9) : 328-330. Northern anthracnose of red clover in Illi¬ nois in 1946. Ibid. 30(9): 330-332. Late blight in Illinois. Plant Disease Re¬ porter Suppl. 165: 334.

Late blight of potatoes in 1946. Illinois. Plant Disease Reporter 30(12): 452.

1947

Gibberella zeae damage in Illinois in 1946 (with Benjamin Koehler). Ibid. 31(4) : 169-170.

Grape downy mildew in Illinois in 1947. Ibid. 31(10) : 391.

1948

Bacterial canker of cowpea in Illinois. Ibid. 32(6): 275.

1949

Late season incidence of Stewart’s disease on sweet corn and winter temperatures in Illinois, 1944-1948. Ibid. 33(4): 192-194.

Bacterial stalk rot of corn in Illinois. Ibid. 33(9) : 342-343.

Rosen’s bacterial stalk rot vs. Elliott’s pythium stalk rot of corn. Ibid. 33(11) : 441.

1950

Stewart’s disease prospect for 1950. Ibid. 34(5): 155.

1952

Stewart’s disease prospects in Illinois for 1952. Ibid. 36(6) : 238.

1953

Stewart’s disease prospects for 1953. Ibid. 37(5): 311-312.

Early season disease of grain crops and alfalfa present an unusual picture in southern Illinois. Ibid. 37(7): 411-412.

1954

Stewart’s disease prospects in Illinois for 1954. Ibid. 38(6): 388.

Sycamore anthracnose severe in Illinois (with R. J. Campana and I. R. Schnei¬ der). Ibid. 38(8): 597-598.

1955

Stewart’s disease prospects for 1955 in Illinois. Ibid. 39(5): 384-385.

1956

Stewart’s disease prospects for 1956. Ibid. 40(5): 367-368.

1957

New diseases for Illinois (with W. L.

Klarman). Ibid. 41(10): 904.

Causes of corn stalk rot in Illinois (with Benjamin Koehler). Ibid. 41(6): 501- 504

1958

Downy mildew on cucurbits in Illinois in 1957. Ibid. 42(4): 544.

Losses caused by crown rust of oats in 1956 and 1957 (with R. M. Endo). Ibid. 42(10): 1126-1128.

1960

Stewart’s disease : expected development in Illinois in 1960. Ibid. 44(5) : 372. Diseases of wheat, oats, barley, and rye. Ill. Nat. Hist. Surv. Circ. 48. 157 p.

1961

A new species of Cercospora on Acer sac- charinum (with Anthony E. Liberta). Mycologia 52(2) : 345-347.

Stewart’s disease: expected development on corn in Illinois in 1961. Plant Disease Reporter 45(5): 393.

1963

Contributions from Illinois (with C. M. Brown and H. Jedlinski). 1962 Oats Newsletter 13: 37-39.

Host plants of charcoal rot disease in Illi¬ nois. Plant Disease Reporter 47(8): 753-755.

1964

Some plant diseases new to Illinois. Ibid. 48(11): 866-870.

Manuscript received January 25, 1971

A LEUCISTIC LITTLE BROWN BAT ( MYOTIS L. LUCIFUGUS)

HARLAN D. WALLEY Department of Biology,

Northern Illinois University, DeKalh, Illinois 60115

Abstract.— Review of the known records of albinism and leucism in Chiroptera, with remarks on a specimen of leucistic Little Brown Bat ( Myotis 1. lucifugus ) from Illinois.

Few records of albinism and leucism of bats have been recorded. Allen (1940) and Setzer (1950) in reviewing the liter¬ ature cited few instances, listing species of only nine genera. Since that time only

twenty-three reports of albinism and leucism in bats have been published. This is a small number when one considers several million bats having been handled during banding operations (Table 1).

While recording recoveries of bats at Blackball Mine, 1.75 miles West of Utica, LaSalle Co., Illinois, on 9 January 1971, an unusual leucistic male, Little Brown Bat, ( Myotis l. lucifugus) was taken from a cluster of 107. The aberrant individual

Table 1.- — Records of albinistic and leucistic traits cited in bats.

Numbers represent individuals.

Species

Albinism

Leucism

Author

Nycteris nana .

1

Verschuren (1955)

Rhinolophus euryale .

1

Dorst (1957)

Anoura caudifera .

1

Linares (1967)

Glossophaga longirostris .

1

Setzer (1950)

Antrozous p. pallidus .

1

Setzer (1950)

Barbastella barbastellus .

Numerous

Palasthy (1968)

Eptesicus capensis .

1

Allen (1940)

Lasiurus borealis .

2

Allen (1940)

Hamilton-Smith (1968)

Dorst (1957)

Miniopterus schreibersi .

1

1

Myotis daubentoni .

1

Haensel (1968)

Myotis grisescens .

3

Tuttle (1961)

Myotis lucifugus .

1

1

Dubkin (1952) and this report

Myotis sodalis .

Numerous

Metzger (1956)

Barbour and Davis (1970)

Myotis velifer incautus .

1

Rogers (1965)

Nyctalus noctula .

1

Dulic and Mikuska (1968)

Nycticeius burner alis .

1

Easterla and Watkins (1968)

Pipistrellus pipistrellus .

1

Allen (1940)

Pipistrellus s. subflavus .

1

Blair (1948)

Mollossus fortis .

1

Heatwole et al. (1964)

Molossus tropidorhynchus .

1

Allen (1940)

Tadarida b. mexicana .

1

Numerous

McCoy (1960) and Glass (1954), Herreid and Davis (1960)

Tadarida ( Chaerephon ) plicatus . .

3

Allen (1940)

Tadarida femorosacca .

1

Mitchell (1963)

[196]

Xotcs

197

(Figure 1) exhibited a completely snow white ventral coloration, with an extensive white patch extending up over the right shoulder and covering approximately half the dorsal surface of the body. The in- terfemoral membrane, head, and wings are of normal pigmentation. The eyes of this individual are black. The speci¬ men is preserved as a skin and skull (HDW 1102). Standard measurements (mm) 80; 33; 10; 14.

I have been unable to find any similar reports for Myotis I. lucifugus, although Dubkin (1952) cites a completely albinistic specimen. Of some 20,000 M. lucifugus handled in northcentral Illinois, only one individual exhibited this trait.

It is interesting to note that leucism occurs rather frequently in three species, Tadarida b r asilie nsis mexicana (Glass, 1954); Barbastella barbastellus (Palasthy, 1968) and Myotis sodalis (Barbour & Davis, 1970).

Figure 1. Leucistic Little Brown Bat ( Myotis l. lucifugus) .

Literature Cited

Allen, G. M. 1940. Bats. Dover Publ. 1-368, figs. 1-57.

Barbour, R. W. and W. H. Davis. 1970. Bats of America. Univ. of Kentucky Press, 1-286, figs. 1-131.

Blair, W. F. 1948. A color pattern aber¬ ration in Pipistrellus subflavus subflavus. J. Mammal. 29: 178-179.

Dorst, J. 1957a. Coloration anormale chez un Minioptere de Schreibers. Mam¬ malia 21 ( 2 ) : 191.

- . 1957. b. L'ncas d'albinisme

complet chez un Rhinolophe euryale. Mammalia 21: 306.

Dubkin, L. 1952. The White Lady.

London ( MacMillan ) .

Dulic, B. and J. Mikuska. 1968. A White Nyctalus noctula (Schreber, 1774). Saugt. Mitt. 16: 308-309.

Easterla, D. A. and L. C. Watkins. 1968. An aberrant evening bat. South¬ west. Nat. 13 (4) : 447-448, 1 fig. Glass, B. P. 1954. Aberrant coloration in Tadarida mexicana. Amer. Midi. Nat. 52 (2) : 400-402, pi. 1.

Haensel, J. 1968. A partial albino waterbat ( Myotis daubentoni ) found in the Rudersdorf chalk quarrv. Milu 2 : 350-354.

Hamilton-Smith, E. 1968. Albinism in the Bent-winged bat ( Miniopterus schrei- bersii (Kuhl). Viet. Nat. 85: 358-359. Pi- L

Heatwole, H., et al. 1964 Albinism in the bat, Molossus fortis. J. Mammal, 45:476. Harreid, C. F. H. and Davis, R. B. 1960. Frequency and placement of white fur on Free-tailed Bats. J. Mammal., 41:117- 119.

Linares. O. J. 1967 Albinism in the long-tongued bat. Anoura caudifera. J. Mammal. 48 (3) : 464-5.

McCoy, C. J. 1960. Albinism in Ta¬ darida. J. Mammal. 41 (1): 119. Metzger, B. 1956. Partial albinism in Myotis sodalis. J. Mammal. 37 (4) : 546.

Mitchell, H. A. 1963. Aberrant white fur in the Pocketed Free-tailed Bat. J. Mammal., 44:422.

Palasthy, J. 1968. The common oc¬ currence of partial albinism in Barbas¬ tella barbastellus Schreber. 1774. Bio- logia 23: 370-376.

Rogers, G. C. 1965. Aberrant coloration in Myotis velifer incautus. Southwest, Nat., 10:311.

Setzer, H. W. 1950. Albinism in bats.

J. Mammal. 31 (3) : 350.

Tuttle, M. D. 1961. Notes and Corre¬ spondence. Bat Banding News 2 (2) : 16.

Verschuren, J. 1955. Un cas d'albi¬ nisme complet chez un cheiroptere, Nycteris nana (A.). Bull. Mus. Hist, nat. Belg. 31 (34) : 1-4.

Manuscript received February 1 . 1971

THE FIRST RECORD IN ILLINOIS OF A POPULATION OF STETHAULAX MABMOBATUS (SAY) (HEMIPTERA: SCUTELLERID AE ) WITH INFORMATION ON LIFE HISTORY

j. e. McPherson and j. f. walt

Department of Zoology ,

Southern Illinois University, Carbondale 62901

Abstract. — The first record in Illinois of a population of Stethaulax marmoratus , and life history information are presented.

The genus Stethaulax contains at least one species in America north of Mexico, S. marmoratus (Say). However, Aula- costethus simulans Uhler, found in Arizona and California, may be a second species of Stethaulax (Blatchley, 1926).

S. marmoratus has been collected in several states including New York, New Jersey, Maryland. North Carolina, Georgia, Missouri, and Texas. Hart (1919) did not list the species for Illinois. However Malloch, in editing Hart’s posthumous pub¬ lication, added that he had collected one female specimen at Cobden, Union Co., May 9, 1918. Since only one specimen has been listed for the state, there is some doubt whether or not the species is es¬ tablished in Illinois.

Little is known of the biology of this insect. It has been collected from cedar (species unknown) and Rhus aromatica Ait. It has also been found on R. cana¬ densis Marsh. (R. aromatica?) . The adult

is mottled in appearance (Fig. 1), meas¬ uring 4. 5-5. 5 mm in width, 6. 0-7. 5 mm in length (Blatchley, 1926; Froeschner, 1941).

On October 17, 1970, we found 23 in¬ dividuals of this species feeding on drupes of R. glabra L. in Jackson Co. on and near the Southern Illinois University cam¬ pus, Carbondale. These sites were approxi¬ mately 14 miles north of Malloch’s site of collection. The animals were well camouflaged and thus, difficult to detect (Fig. 2).

There are five nymphal instars as de¬ termined by rearing in the laboratory. In the field, evidently, the entire develop¬ mental period is spent on the drupes. Eggs and first-instar nymphs were not col¬ lected. However, egg shells found on Oc¬ tober 31 indicated the eggs had been laid in clusters, typical of scutellerids studied thus far, on the twigs and surrounding drupes. It is very probable, based on work previously done on related species, that the first-instar nymphs are also found here.

Second-instar nymphs through adults were found continuously October 17-31, actively feeding. On October 31, approxi-

Figure 1.

Female (left) and male of Stethaulax

marmoratus in copulo.

[198]

Notes

199

mately 80% of the 81 individuals collected were adults. By mid-November, the in¬ sects had left the host plants.

The length and width of 10 adult males and females were measured (Table 1). Length was from tip of tylus to hind abdominal margin; width was across the humeral angles. Measurements approxi¬ mated those previously reported. Also, using Student’s t-test, the lengths and widths of females were found to be sig¬ nificantly larger than those of males (P < .025).

Our collection dates are unique because specimens taken by other investigators have been between May 7 and September 16. Also, it was unusual to find individuals still developing at this time since, to our knowledge, individuals of other species of Scutelleroidea in this area had completed development by the end of September of the same year. It is impossible to state whether this species is uni- or bivoltine.

Based on the above information, this species appears to be well established in southern Illinois.

Table 1. — Measurements of adult Stethaulax marmot atus in mm.

Male

Female

Length

Width

Length

Width

Number .

x + S.E .

Range .

10

6.7 + .2

5. 7-7. 3

10

4.5 + .1

3. 8-4. 9

10

7.6 + .1

7. 1-8.3

10

4.9 + .1

4. 4-5. 2

Figure 2. Two fifth-instar nymphs hidden among drupes of Rhus glabra.

200

Transactions Illinois Academy of Science

Acknoweldgments

We wish to thank Dr. H. I. Fisher, Southern Illinois University, for reviewing the manuscript and Dr. R. G. Froeschner, U. S. National Museum, for confirming our identification of this insect. We also are grateful to Mr. John A. Richardson, Southern Illinois University, who provided the photographs.

References Cited

Blatchley, W. S. 1926. Heteroptera or true bugs of eastern North America with

especial reference to the faunas of In¬ diana and Florida. Nature Publ. Co., Indianapolis. 1116 p.

Froeschner, R. C. 1941. Contributions to a synopsis of the Hemiptera of Mis¬ souri. Part 1, Scutelleridae, Podopidae, Pentatomidae, Cydnidae, Thyreocoridae. Amer. Midi. Nat. 26:122-146.

Hart, C. A. 1919. The Pentatomoidea of Illinois with keys to the nearctic ge¬ nera. Illinois Nat. Hist. Surv. Bull. 13 : 157-223. (edited by J. R. Malloch) .

Manuscript received February 2, 1971

MR. STOVER

HIRAM F. THUT AND JOHN EBINGER Department of Botany ,

Eastern Illinois University, Charleston, Illinois

Professor Ernest L. Stover, died Novem¬ ber 30, 1969. He taught at Eastern Illi¬ nois College and University for 37 years, was a past president of the Illinois State Academy of Science, served on its council for ten years, and served on various com¬ mittees concerned with teaching.

Mr. Stover, the son of a Methodist min¬ ister, was born in Belbrooke, Ohio, on August 28, 1893. He grew up in southwes- ern Ohio, attended elementary school in Jeffersonville and Mechanicsburg and grad¬ uated from the Richwood High School. After a year at Adrian College, Mr. Stover finished his bachelor’s degree at Ohio State in 1917.

As a college student, Mr. Stover played a baritone horn in the marching band. World War I interrupted his graduate work. He served with the Engineers in 1918 and saw limited action. Soon after the Armistice, he asked to be transferred to the band as an entertainer. For some months in 1919 he was stationed in Paris, entertaining soldiers. Recently Mr. Stover received his 50th year citation as a Legion¬ naire. He also had the opportunity to study briefly at the Sorbonne and the Pasteur In- stite. His study at the Pasteur Institute made him keenly aware of the opportunities in science, especially botany. While in Paris, he bought a violoncello and took private music lessons. This violoncello was a pride and Joy to Mr. Stover for the rest of his life.

On returning to the states, Mr. Stover became an instructor in botany at the Ohio State University, 1919-1923. At that time he was not sure whether it would be botany or music. He was playing cello quite suc¬ cessfully with a theatre orchestra and some hotel bands. However, in time he was influenced by Dr. E. N. Transeau to give botany his primary effort. He obtained his Master’s degree in botany in 1921. That was followed by graduate work at the University of Chicago, where he ob¬ tained the Ph.D. degree in botany in 1924.

Mr. Lord, President at Eastern, needing a botanist, asked his very good friend Dr.

Transeau, who had taught at Eastern from 1908-1915, to recommend a likely candi¬ date for the position. Dr. Transeau rec¬ ommended the man he had persuaded a couple of years earlier to become a botan¬ ist. In September 1923 Mr. Stover joined the faculty of the Eastern Illinois State Teacher’s College at Charleston. Mr. Stover served as botanist and head of the department until 1960.

Mr. Stover, as a botanist, had two pri¬ mary interests — good teaching and plant anatomy. His 30 some odd scientific ar¬ ticles and books were published almost alternately between teaching and plant anatomy. As a teacher Mr. Stover want¬ ed his students to know plants. He had a way of making a student feel that they had discovered something together while being helped through the recognition keys. Early in his teaching career he affiliated himself with the Illinois Academy of Sci¬ ence and served on several committees. His particular interest was in the committees that had to do with teaching.

The Botanical Society of America be¬ came much interested in good teaching, and in 1936 appointed a committee on the teaching of botany in colleges and uni¬ versities. Mr. Stover was appointed chair¬ man of that committee and with the help of a grant from the General Education Board and the research assistance of Dr. Clark W. Horton, they published two bul¬ letins. In 1941 he was chairman of a committee on instruction in the biological sciences for the National Research Council.

Besides his botanical work, Mr. Stover had many other interests. He was a mem¬ ber of the National Education Association, a member of the Illinois Education Asso¬ ciation and was President of the Eastern Division of the Illinois Education Associa¬ tion in 1943. He held membership in the A.A.A.S. (fellow), in Sigma Xi and Gam¬ ma Alpha. He retained his interest in music and was a member of Eastern’s Col¬ lege Orchestra for years.

Mr. Stover married Patsy H. Lupo in

[201]

202

Transactions Illinois Academy of Science

1923. They had been graduate students together at the University of Chicago and later she was teaching botany at Rockford College. She died in 1956. In 1959 he married Ethel Hanson, a music instructor at Eastern.

His death removed from our midst a scholar, a skilled teacher, a sensitive per¬ son, a successful administrator and most of all a friend. That was Mr. Stover.

List of Publications

The vascular anatomy of Calamovilfa longi- folia. Ohio Journ. Sci. 24:169-178. 1924.

Trees and shrubs of the campus. Eastern Illinois State Teachers College Bull. 89:1-39. 1925.

The roots of wild rice. Zizania aquatica L. Ohio Jour. Sci. 28:43-49. 1929.

A method of staining microscopic slides for beginning students. Trans. Ill. St. Acad. Sci. 21:186. 1929.

A method of preserving the natural color of certain fungi. Trans. Ill. St. Acad. Sci. 21:187. 1929.

A mesophytic ravine “Rocky Branch” a floristic account. Eastern Illinois State Teachers College Bull. 110:1-26. 1930.

Training teachers in the Biological Sciences.

Ill. Teacher. 20:45-47, 68. 1931.

Trees and shrubs of the campus. 1st. rev. print. Eastern Illinois State Teachers College Bull. 117:1-48. 1932.

Life history of Nymphoides peltatum. Bot.

Gaz. 93:474-483. 1932.

The development and differentiation of tis¬ sues in the stems of grasses, (abstract). Trans. Ill. St. Acad. Sci. 25:130. 1933. Development and differentiation of tissue in the stem tips of grasses. Ohio Jour. Sci. 34:150-160. 1934.

Science in the Elementary School and High School. Ill. Teacher. 23:239, 268-270. 1935.

The need for more than adequate train¬ ing for science teachers. Ill. Teacher. 23:289-290, 304-306. 1935.

A simple apparatus for the steam method of softening woods for microscopic sec¬ tions. Trans. Ill. St. Acad. Sci. 28:87. 1935. (with G. E. Davis).

Collection of fleshy Ascomycetes from East Central Illinois. Trans. Ill. St. Acad. Sci. 28:95-96. 1935. (with lea Marks) . An interesting preservation of color in the

algae and certain fungi. Trans. Ill. St. Acad. Sci. 29:76. 1936.

The embryo sac of Eragrostis cilianensis (All.) Link: A new type embryo sac and a summary of grass embryo sac in¬ vestigations. Ohio Jour. Sci. 37:172- 184. 1937.

Regions of growth in hypocotyls. Trans. Ill. St. Acad. Sci. 30:105-106. 1937.

(with C. E. Brian).

Telling the truth in Elementary Science. School Science and Mathematics. 37: 665-666. 1937.

College grades in the biological sciences as related to Secondary School training. Trans. Ill. St. Acad. Sci. 31:115-116. 1938. (with H. F. Thut).

An exploratory study of the teaching of botany in the colleges and universities of the United States. Bot. Soc. Amer. Publ. 119:1-46. 1938. (with others).

Achievement tests in relation to teaching objectives in general college botany. Bot. Soc. Amer. Publ. 120:1-71. 1939.

(with others).

A collection of Myxomycetes from Eastern Illinois. Trans. Ill. St. Acad. Sci. 34:95. 1941.

Adjustment of the college curriculum to wartime conditions and needs. U. S. Office of Education, Biological Science Report No. 19. 1943. (with others).

Varying structure of Conifer leaves in dif¬ ferent habitats. Bot. Gaz. 106:12-25. 1944.

The objective results of different teaching methods in general Botany. Educational Administration and Supervision. 31: 513-529. 1945. (with W. M. Gilbert).

A key to the genera of common woods. Trans. Ill. St. Acad. Sci. 39:65-66. 1946.

The men of agriculture, our first citizens.

School and Society. 65:289-291. 1947. Mark Hopkins and the modern log. Trans.

Ill. St. Acad. Sci. 41:6-12. 1948.

An introduction to the anatomy of seed plants. D. C. Heath and Co., Boston, xiv + 274 pp. 1951.

Trees and shrubs of the campus and East Central Illinois. Eastern Illinois State College Bull. 200:1-76. 1952.

Question and answer book in general bot¬ any. Wilcox and Follett. 1952. Graduate Students in Botany. PI. Sci. Bull. 7(4): 1-3. 1961.

Manuscript Received December 21 , 1970

Contributing Members of 1970 Roger Adams Joan O. Dunn James R. Getz Virgil Dean Winkler Joseph A. Beatty Harold Walter Bretz Mrs. Charles C. Colby Ronald L. Cook Jerry H. Elliston Robert H. Herman George H. Otto Raymond E. Reed Harry Sicher Dr. Samuel J. Turner Milton D. Thompson Ted F. Andrews James Edward Palmer Richard S. Jackson, Jr.

James Ferry, Jr.

[203]

New York Botanical Garden Library

3 5

85 00341 4792

TRANSACTIONS of the ILLINOIS STATE ACADEMY OF SCIENCE

Editorial Board:

Malcolm T. Jollie, Northern Illinois University, Editor and Chairman

Stanley H. Frost, Northern Illinois University, Geology Gordon C. Kresheck, Northern Illinois University, Chemistry John C. Shaffer, Northern Illinois University, Physics Darrell L. Lynch, Northern Illinois University, Microbiology Robert H. Mohlenbrock, Southern Illinois University, Botany

Articles in the Transactions pertaining to Biology, Chemistry, and Geology are abstracted in Biological Abstracts, Chemical Abstracts, and Abstracts of North American Geology.

The current Transactions may be obtained by payment of annual dues.

Exchanges may be arranged or previous issues may be purchased by addressing Paul Parmalee, Illinois State Museum, Springfield, Ill. 62706.

Mailing date, July, 1971.

jCanc£of£inco6v

Transactions

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3

of the

Illinois

State Academy

of Science

LIBRARY

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NEW YORi< BOTANICAL GARDEN4

Volume 64 No. 3 1971

TISAAH

TRANSACTIONS of the ILLINOIS STATE ACADEMY OP SCIENCE

Editorial Board:

Malcolm T. Jollie, Northern Illinois University, Editor and Chairman

Stanley H. Frost, Northern Illinois University, Geology Gordon C. Kresheck, Northern Illinois University, Chemistry John C. Shaffer, Northern Illinois University, Physics Darrell L. Lynch, Northern Illinois University, Microbiology Robert H. Mohlenbrock, Southern Illinois University, Botany

Articles in the Transactions pertaining to Biology, Chemistry, and Geology are abstracted in Biological Abstracts, Chemical Abstracts, and Abstracts of North American Geology.

The current Transactions may be obtained by payment of annual dues.

Exchanges may be arranged or previous issues may be purchased by addressing Paul Parmalee, Illinois State Museum, Springfield, Ill. 62706.

Mailing date Feb., 1972

34762—1650—9/71

TRANSACTIONS

OF THE

ILLINOIS STATE ACADEMY OF SCIENCE

VOLUME 64 - 1971

No. 3

Illinois State Academy of Science

AFFILIATED WITH THE

Illinois State Museum Division Springfield, Illinois

(PRINTED BY AUTHORITY OF THE STATE OF ILLINOIS) Richard B. Ogilvie, Governor

December 1, 1970

CONTENTS

Cardiac Output and Central Blood Volume as a Function of Body Weight in the Baboon

By G. S. Moss, L. D. Homer, C. M. Herman, Donald Siegel, Alan

Cochin, and Vidal Fresquez . 207

Evaluation of the Digestion — Baermann Technique for the Detection of Dead Trichnae

By William G. Dyer and Valerie A. Evje . 212

Symposium: A New Role for the Academy — University of Illinois, Chicago Circle Campus, April 24, 1970:

Symposium Introduction

By A. J. Pappelis . 215

A New Role for the Illinois Academy of Science

By SENATOR Alan J. Dixon . 217

Bryophytes of Goose Lake Prairie, Illinois

By William M. Zales . 222

Fisher and Porcupine Remains from Cave Deposits in Missouri

By Paul W. Parmalee . 225

Cultivation, Life History and Salinity Tolerance of the Tidepool Copepod,

Tigriopus Californicus Baker 1912, in Artificial Sea Water

By Harry W. Huizinga . 230

Incidence of Mercury in Illinois Pheasants

By William L. Anderson and Peggy L. Stewart . 237

Longitudinal Pattern of Nuclear Size in Bulb Scale Epidermis of Allium cepa and Changes in Size in Response to Neckrot

By F. B. Kulfinski and A. J. Pappelis . 242

Versatile Apparatus for Studying Reactions Involving Gas Adsorption or Evolution

By Josephus Thomas, Jr., and Robert R. Frost . 248

Catalog of Paleozoic Paleozoological Type and Figured Specimens at the Illinois State Museum

By Richard L. Leary . 254

The Cyperaceae of Illinois. XII. Carex, Part 1

By Dan K. Evans and Robert H. Mohlenbrock . 260

Gastric Morphology in Selected Mormoopid and Glossophagine Bats as Related to Systematic Problems

By G. Lawrence Forman . 273

Studies on the Phenol-soluble Lipopolysaccharides From Serratia marcescens Bizio

By Joseph C. Tsang . 283

The Response of Southern Illinois Barren Vegetation to Prescribed Burning

By Roger C. Anderson and John Schwegman . 287

Analysis of the Electron Density and Potential of Solid Benzene

By J. L. Amoros and Marisa Canut-Amoros . 292

Plant Investigations II. Studies on the Hexane Extract of Cirsium arvense

By David M. Piatak and Larry S. Eichmeier . 300

NOTES:

Longevity Record for Pipistrellus subflavus

By Harlan D. Walley and William L. Jarvis . 305

Notes on Illinois and Wisconsin Resupinate Basidiomycetes By Anthony E. Liberta . 306

Morris M. Leighton (1887-1971)

By Boris Musulin . 307

Gilbert H. Cady (1882-1970)

By Boris Musulin . 308

CARDIAC OUTPUT AND CENTRAL BLOOD VOLUME AS A FUNCTION OF BODY WEIGHT IN THE BABOON

G. S. MOSS, L. D. HOMER, C. M. HERMAN, DONALD SIEGEL, ALAN COCHIN, AND VIDAL FRESQUEZ

Department of Surgery, University of Illinois College of Medicine and Hektoen Institute for Medical Research, Cook County Hospital, Chicago, Illinois 60612

Abstract. — The relationship between cardiac output and central blood volume as a function of body weight was investi¬ gated in tranquilized adult baboons. Car¬ diac output was determined by the dye dilution method. Central blood volume was calculated as the product of cardiac output and mean transit time. It was concluded that weight raised to the .62 power offers a slight improvement in com¬ paring cardiac outputs between animals of different sizes. A simple weight index for central blood volume should be a satisfactory expression for the parameter.

To compare changes in cardiac output and central blood volume observed in baboons with those ob¬ served in other animals of varying sizes, it would be helpful to have an index for the basal value of cardiac output and central blood volume in the baboon, based on weight or some function of weight. For the purposes of this report, Central Blood Volume (CBV) is defined as that volume of blood contained be¬ tween the tips of the sampling cath¬ eters in the right atrium and the arch of the aorta. An examination of the relationship between weight and these variables in the baboon is the subject of this report.

Method

Adult baboons ( Papio doguera) weighing between 13.7 and 27.5 kg were used in this study. The basal cardiac output in 48 animals and the central blood volume in 19 were determined according to a tech¬ nique published elsewhere (Moss et. al., 1968). Briefly, in each ani¬ mal, a siliconized heparin-filled plas¬ tic catheter was implanted in the thoracic aorta and another in the pulmonary artery through a left

thoracotomy two to three weeks prior to study. The ligated free ends of the two catheters were implanted under the skin of the left chest and the animal was then returned to his cage. On the morning of the study, the baboon was tranquilized with l-(phenylcyclohex) piperidine HCL (Sernylan, Parke-Davis), 1 mg/kg of body weight, based on the ani¬ mal's original weight. The animal was then reweighed and the second weight was used for this report. Each animal was then placed in the prone position, with its anterior chest wall protected by padded ax¬ illary supports and the extremities loosely restrained.

After a one-hour basal period, two dye dilution curves were ob¬ tained in the following fashion. Five milligrams (1 ml) of indocyanine green dye were injected into the pulmonary artery catheter, followed by a forceful 7 ml bolus of isotonic saline. Arterial blood from the aor¬ tic catheter was simultaneously as¬ pirated through the cuvette of a cardiodensitometer (Beckman) at a rate of approximately 20 ml/min by means of a withdrawal pump (Gilford, Model 105-S). The cardio¬ densitometer recorded the resultant indicator dilution curve and also mechanically integrated the area under the curve, including recircu¬ lation. Correction for recirculation was made by extrapolating the ex¬ ponential downslope to the base line, or, in most cases, by using a logarithmic nomogram provided by the manufacturer for this purpose. Central blood volume was calcula¬ ted as the product of cardiac out-

207

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put and mean transit time (Hamil¬ ton et. al., 1932, Kinsman et. al., 1929, Stewart, 1921-22). Mean tran¬ sit time was obtained from the least squares fit of a gamma variate to the dye curves (Thompson et. al., 1964).

Statistical Analysis: The relation¬ ship between the average of the two cardiac output measurements and weight was examined by the least square method after taking the nat¬ ural logarithms of both sets of num¬ bers. Central blood volume versus weight was examined in a similar fashion. The general formula for this regression is log Y = bo + bi . W, where Y = either cardiac out¬ put or central blood volume, bi = slope, bo = intercept, and W = weight. The 95% confidence limits were calculated with a table of t.

The estimating equation for car¬ diac output was log C. 0. = -0.70 + 0.62 log weight. The standard error of the coefficient of log weight was 0.21.

The estimating equation for cen¬ tral blood volume was CBV = 1.80 + 1.19 log weight. The standard error of the coefficient of weight was 0.44.

Results

Figure 1 shows the scatter dia¬ gram of cardiac output and weight plotted on a double logarithmic scale. Also show'n in the diagram is the regression line fitted by least squares and the 95% confidence limits for Y. The estimating equa¬ tion for the best fit is log cardiac output = -0.70 + 0.62 log weight. Although the relationship between cardiac output and weight is sta¬ tistically significant (p<0.01), it is at best only approximate. This is evidenced by a low coefficient of correlation, R = 0.39; wide 95% confidence limits of b1, 0.19 to 1.04; and wide 95% confidence limits of bo, 0.57 to - 1.97.

Figure 1. The relationship between cardiac output and weight in 48 tran- quilized baboons. SEbo = the standard error of the intercept, and SE bi = the standard error of the slope.

The mean cardiac output (liters/ minute) for the 48 baboons was 3.80 ± 0.82. The coefficient of vari¬ ation was 26%. The index obtained when the cardiac output of each animal was divided through by (weight)0-62 had a mean value of 0.50 ± 0.10. The coefficient of vari¬ ation was 20%. The reduction in the coefficient of variation gained in the latter expression is only a minor improvement in expressing cardiac output.

Figure 2 shows the relationship between CBV and weight on a dou¬ ble logarithmic plot. Also shown are the least squares line, 95% confi¬ dence limits of Y, standard error of the slope and intercept, and the coefficient of determination. De¬ spite a reasonably high R value, there is a substantial degree of scat-

CENTRAL BLOOD VOLUME (ml )

Moss etal — Central Blood Volume of Baboon

209

Figure 2. The relationship between central blood volume and weight in 19 adult tranquilized baboons. SEbo = the standard error of the intercept, and SE bi = the standard error of the slope.

ter in the relationship. The slope of the line is not significantly dif¬ ferent from 1.0. The central blood volume index for the 19 baboons is 10.7 ± 2.9 ml/kg.

Discussion

A number of investigators have studied the relationship between cardiac output and some function of body weight in animals and man (Brotmacher et. al., 1956, Cour- nand et al., 1945, Stead et al., 1945, Tanner, 1949, Taylor et al., 1952). Most have reported a significant but low correlation, as found in this study. A high degree of scatter was observed consistently. Some have advocated using body weight raised to the two-thirds power, al¬ though others have favored the three-fourths power. In this study of baboons, cardiac output was best

correlated with weight raised to 0.62 power, which is quite close to the two-thirds power. However, be¬ cause of the large standard error of the exponent, either number could have been selected in calcu¬ lating an expression for cardiac out¬ put for the baboon, based on weight. Indeed, it is not clear that either method is superior to simply divid¬ ing cardiac output by weight.

Guyton (1963) recalculated a si¬ milar expression for animals of all sizes, from the rat to the horse, us¬ ing weight raised to the two-thirds power. He found that in general the value for cardiac output varied directly with body weight. The rat cardiac output was 0.15 L/kg2/3, the dog cardiac output was 0.31 L/kg2/3, and the horse cardiac out¬ put was 0.46 L/kg2/3. Our reported baboon cardiac output of 0.50 L/kg0-62 is consistent with this gen¬ eral scheme in that it lies between the values for rats and horses and not far from that of the dog.

For making rough comparisons between the results in baboons and other animals of varying sizes, the baboon cardiac output divided by (weight)0-62 may prove helpful. How¬ ever, in baboons of a similar size to those studied in our investiga¬ tion, the simple value for cardiac output alone or cardiac output di¬ vided by weight should be satisfac¬ tory for comparison.

Abel, Waldhausen, Daly and Pearce (1967), in a study of hemor¬ rhagic shock in five young baboons weighing 8.8 to 10.5 kg. reported basal cardiac outputs of 84 ml/min per kilogram. This figure is sub¬ stantially lower than the 150 ml/ min per kilogram we would calcu¬ late for a 20 kg. baboon in our study, and well below the 95% confidence region of our estimating equation extrapolated to 10 kg. The upper 95% limit of these investigators was about 140 ml/kg.

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The basal cardiac output of the dogs in the study of Abel et ah, (1967), when adjusted to the two- thirds power of body weight after the manner of Guyton, was 0.26 1/kg2/3. For the baboons in their study, the figure would be approxi¬ mately 0.17 1/kg2/3.

We do not know why our esti¬ mate of cardiac output differed from that of Abel and co-workers. We did note, however, that their animals were referred to as “y°uns” and that their anesthetic consisted of a mixture of barbiturates, given both intramuscularly and intravenously, together with morphine given intra¬ muscularly. In contrast to the dis¬ parity in cardiac output estimates, their CBV estimate of 10.2 ml/kg closely agreed with our estimate.

The validity of expressing cen¬ tral blood volume as the product of cardiac output and mean transit time was confirmed by Schlant and his associates (Schlant et al., 1959). They compared this measurement with results observed when the ac¬ tual amount of blood in the heart and lungs was calculated from the total radioactivity in these organs in dogs previously treated with chromium-51 labeled red blood cells. They found a high correlation (R = + 0.88).

Since the slope of the line shown in Figure 2 is not significantly dif¬ ferent from 1.0, dividing central blood volume estimates by body weight should provide a means of reporting this variable that would make approximate comparisons with other baboons convenient. It is not at all certain, however, that the same index would be suitable for comparison with other species. If, in fact, CBV increases linearly with weight, while cardiac output does not, then the mean pulmonary transit time of indicator should be greater in animals of large species than in those of small species.

Acknowledgements

This study was supported by Research Task No. MR005. 19-0004, from the Bu¬ reau of Medicine and Surgery, U.S. Navy Department. The opinions or assertions contained herein are the private ones of the authors and are not to be construed as offiicial or as reflecting the views of the Navy Department or of the Naval Ser¬ vice at large.

References

Abel, F. L., Waldhausen, J. A., Daly, W. J., and Pearce, W. L. 1967. Pul¬ monary blood volume in hemorrhagic shock in the dog and primate. Amer. J. Physiol., 213:1072.

Brotmacher, L., and Deuchar, D. C. 1956. The systemic blood flow in con¬ genital heart disease, with an examina¬ tion of the validity of the cardiac in¬ dex. Clin. Sci., 15:441.

Cournand, A., Riley, R. L., Breed, E. S., Baldwin, E., and Richards, D. W. 1945. Measurement of cardiac output in man using technique of catheteriza¬ tion of right auricle or ventricle. J. Clin. Invest., 24:106.

Guyton, A. C. 1963. Circulation Physiol¬ ogy: Cardiac Output and Its Regula¬ tion, W. B. Saunders, Philadelphia and London p. 468.

Hamilton, W. F., More, J. W., Kins¬ man, J. M., and Spurling, R. G. 1937. Studies on circulation; further analysis of injection method, and of changes in hemodynamics under physiological and pathological conditions. Amer. J. Phy¬ siol., 99:537.

Kinsman, J. M., More, J. W., and Ham¬ ilton, W. F. 1929. Studies on circula¬ tion; injection method; physical and mathematical considerations. Amer. J. Physiol., 89:322.

Moss, G. S., Proctor, H. J. Herman, C. M., Homer, L. D., and Litt, B. D. 1968. Hemorrhagic shock in the ba¬ boon. I. Circulatory and metabolic ef¬ fects of dilutional therapy: Preliminary report. J. Trauma, 8:837.

Schlant, R. C., Novack, P., Kuaus, W. L., Moore, C. B., Haynes, F. W., and Dexter, L. 1959. Determination of central blood volume. Comparison of Stewart-Hamilton method with direct measurements in dogs. Amer. J. Phy¬ siol., 196:499.

Stead, E. A., Warren, J. V., Merrill, A. J., and Brannon, E. X. 1945. Car¬ diac output in male subjects as mea¬ sured by technique of right atrial cath¬ eterization. Normal values with obser¬ vations on the effect of anxiety and tilting. J. Clin. Invest., 24:326.

Moss etal — Central Blood Volume of Baboon

211

Stewart, G. N. 1921-22. The pulmonary circulation time, the quantity of blood in the lungs and the output of the heart. Amer. J. Physiol., 58:20.

Tanner, J. M. 1949. Construction of normal standards for cardiac output in man. J. Clin. Invest., 28:567.

Taylor, H. L., and Tiede, K. 1952. The comparison of the estimation of the

basal cardiac output from a linear form¬ ula and the “cardiac index.” J. Clin. Invest., 31:209.

Thompson, H. K., Jr., Starmer, F., Whalen, R. E., McIntosh, H. D. 1964. Indicator transit time considered as a gamma variant. Circulation Res., 14:502.

Manuscript received November 10, 1970

EVALUATION OF THE DIGESTION— BAERMANN TECHNIQUE FOR THE DETECTION OF DEAD

TRICHNAE

WILLIAM G. DYER AND VALERIE A. EVJE

Department of Zoology, Southern Illinois University, Carbondale 62901 and Department of Biology, Minot State College, Minot, North Dakota 58701

Abstract. — The value of the digestion- Baermann technique as a diagnostic tool for the detection of dead Trichinella spira¬ lis larvae was reinvestigated. Quantita¬ tive studies showed this method to be only 6.0 per cent efficient and that as high as 31.0 per cent of the trichinae failed to pass through the opening of the screen. Examination of the filtrate only would have failed to detect a significantly large number of dead larvae. A decrease in the number of larvae recovered appeared to be directly proportional to the length of the digestion period.

It is generally agreed that the digestion - Baermann technique is rather dependable for detecting live larvae of Trichinella spiralis (Owens, 1835) Raillett, 1895. However, in surveying the prevalence of T. spi¬ ralis in wild animal populations it is not always possible to examine fresh tissue and some carcasses must be frozen for inspection at a later period. The efficiency of the diges¬ tion-Baermann technique as a diag¬ nostic tool for detecting dead larvae has for the most part been evalu¬ ated on the results of comparative studies utilizing data obtained from tissue examined both by the direct microscopic method and by the di¬ gestion-Baermann technique. In view of the fact that some workers continue to utilize the digestion- Baermann method only for exam¬ ining either frozen tissue or tissue removed from the dried skins of animals, the purpose of the present investigation was to reevaluate the digestion-Baermann method on the basis of data obtained from studies containing quantitative elements.

Materials and Methods

Dead trichinae were obtained

from the diaphragms and skeletal muscles of experimentally infected rats which had been frozen for two months and subjected to a modi¬ fied artificial digestion-Baermann technique of Kerr et al. (1941) in that a 60-mesh screen and a diges¬ tion period of 2.5 hr were substi¬ tuted for an 80-mesh screen and an 18 hr digestion period. Approxi¬ mately 200 dead larvae were iso¬ lated, examined, transferred to a watch glass, and counted four times. The dead larvae were then added to a trichinae-free 40 gm sample of skeletal muscles of frozen white¬ tailed jack rabbits, Lepus townsendii Backmann, 1839, which had been artificially digested in 2 liters of a mixture of 1.0 per cent pepsin so¬ lution and 0.7 per cent hydrochloric acid at 37° C. for 4 hr under con¬ stant mechanical agitation. After settling for 1 hr, two-thirds of the mixture was siphoned off and ex¬ amined for the presence of larvae. The remaining material was poured through an 80-mesh screen into a 3-liter funnel and enough water added to cover the screen. The fil¬ trate was then drawn into a ruled petri dish for examination and counting of trichinae under a dis¬ secting microscope. The screen was transferred to a large container, in¬ verted, washed several times with a strong stream of water and the wash transferred to a funnel with a short neck closed by means of rub¬ ber tubing and a Hofmann clamp. The wash was then drawn into a ruled petri dish for examination and counting of trichinae.

212

Dyer & Evje — Baermann Technique Evaluation

213

Table I. — Recovery of Trichinella spiralis added to digest rabbit muscle.

Recoveries

Trials

No.

Larvae

Added

Filtrate

No.

Screen

No.

No.

Total

%

1

200

14

53

67

33.5

2

202

12

8

20

9.9

3

198

24

61

85

42.9

4

199

11

53

64

31.8

5

197

9

74

83

41.6

6

200

12

102

114

57.0

7

200

14

56

70

34.8

8

200

6

39

45

22.5

9

200

10

79

89

44.5

10

200

9

91

100

50.0

11

199

13

60

73

36.7

Mean

200

12

62

74

37.0

Results

Examination of the supernatant showed it to be trichinae-free. The results of 11 trials to determine the efficiency of the digestion-Baer- mann technique are shown in Table I. An average of only 12 (6.0 per cent) of 200 larvae added to di¬ gested rabbit muscle was recovered in the filtrate, and 62 larvae (31.0 per cent) failed to pass through the openings in the mesh. An average of 63.0 per cent of the 200 larvae added to digested rabbit muscle were not recovered.

Discussion

Examination of the filtrate only would have failed to detect a sig¬ nificantly large number of dead larvae added to the digested rabbit muscle. In routine procedure the screen would not be examined with the high probability of missing light infections due to the failure of dead larvae to pass through the openings of the screen. Hence, the digestion- Baermann technique is inadequate as a diagnostic procedure for the detection of dead trichinae. These findings agree with those of Hall and Collins (1937, p. 471) who stated that . . . “since the efficiency of the Baermann apparatus de¬ pends for its effect on the move¬ ment of live worms and the effect

of gravity in bringing down these moving worms, the digestion meth¬ od is of little value for the de¬ tection of dead trichinae unless these are present in numbers large enough to insure that some of them will land directly on the screen and fall through. ” Zimmermann et al. (1961) in a study on the occurrence of T. spiralis in pork sausage also em¬ phasized that the digestion-Baer- mann technique is of little value for detecting dead larvae.

Preliminary observations re¬ vealed that the internal organs of a few larvae were ruptured following a digestion period of 2.5 hr and that the number of larvae injured as well as the decrease in the number of larvae recovered appeared to be directly proportional to the length of the digestion period. However, these observations were not quan¬ tified as neither the percentages of larvae injured nor recovered were recorded at this time.

Ransom (1916) demonstrated that artificial digestion for 24 hrs or less had no appreciable effect upon the viability of trichinae. Gursch (1948) studied the recovery of T. spiralis from the digestive system of experimentally infected rats previously exposed from 4 to 12 hr of artificial digestion and 0 to 72 hr of refrigeration (5° C.). Ac-

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Transactions Illinois Academy of Science

cording to Gursch, 4 to 8 hr of diges¬ tion and up to 24 hr storage in the refrigerator did not impair the in- fectivity of the trichinae, but that a 12 hour period of digestion de¬ creased the percentage of worms recovered from test infections. He also found that exposure to 72 hr of refrigeration was definitely inju¬ rious to the larvae even in combi¬ nation with a few hours of diges¬ tion. From the findings of Gursch and the preliminary observations in the present study, it is highly probable that the reverse procedure (exposure to freezing followed by even a short period of digestion) may result in the complete destruc¬ tion of some larvae. These factors may largely account for the low combined recoveries (filtrate and screen) of trichinae utilizing the di- gestion-Baermann technique.

Acknowledgement

This study was supported in part by research grant 2-17-43 of the Southern Illinois University Office of Research and Projects and by an Undergraduate Re¬

search Participation Grant (GY-5851) form the National Science Foundation.

The Authors wish to express sincere thanks to Dr. William J. Zimmermann, Veterinary Medicine Research Institute, Iowa State University for providing the infected rats used in this study.

Literature Cited

Gursch, 0. F. 1948. Effects of digestion and refrigeration on the ability of Tri- chinella spiralis to infect rats. J. Para- sit. 34:394-395.

Hall, M. C., and B. J. Collins. 1937. Studies on trichinosis. I. The incidence of trichinosis as indicated by post¬ mortem examination of 300 dia¬ phragms. Pub. Hlth. Rep. 52:468-490. Kerr, K. B., L. Jacobs, and E. Cuvil- lier. 1941. Studies on trichinosis. XIII. The incidence of human infection with trichinae as indicated by post-mortem examination of 3,000 diaphragms from Washington, D. C., and 5 eastern sea¬ board cities. Pub. Hlth. Rep. 56:836- 855.

Ransom, B. H. 1916. Effects of refrigera¬ tion upon the larvae of Trichinella spi¬ ralis. J. Agric. Res. 5:819-854. Zimmermann, W. J.,L. H. Schwarte, and H. E. Biester. 1961. On the occurrence of Trichinella spiralis in pork sausage available in Iowa (1953-60). J. Parasit. 47:429-432.

Manuscript received February 18, 1971

SYMPOSIUM: A NEW ROLE FOR THE ACADEMY

UNIVERSITY OF ILLINOIS,

CHICAGO CIRCLE CAMPUS,

APRIL 24, 1970

SYMPOSIUM INTRODUCTION A. J. PAPPELIS

Department of Botany, Southern Illinois University, Carbondale, Illinois 62901

During the past two decades, I have formed an opinion that most of my colleagues believe the state academies of science are ineffective organizations for professional ac¬ tivities of senior scientists and for improvement of life in our society. They assign to the academies the roles of being the training ground for graduate students to present research papers; the caretaker of a state-wide high school science club program; a science fair organiza¬ tion; the organizer of a journal that has little, if any, recognized value in professional circles, even within the state; and, the organizer of in¬ effective committees that may dis¬ cuss problems of limited interest and have trouble getting an agree¬ ment on the wording of a resolution to bring before the annual meeting. This view of the state academies changes when persons are asked about the New York Academy of Science. Senior scientists are aware of monograph publications or spe¬ cial topic symposia conducted by that academy. To many of us, the question of defending the Acade¬ mies does not arise, for all of the mentioned activities are essential. The question that does arise is - What else should the Academies be doing?

With regard to the Illinois Acad¬ emy, I have often suggested a break from the traditional annual meet¬ ing and annual field trip. Some changes have occurred under the direction of recent officers. But, as chairman of the Botany section in

1968, 1 found it impossible to change the member’s habits without chang¬ ing the traditional role of the sec¬ tion chairman. A change in the man¬ ner that the annual meetings are arranged appears necessary. The section chairmen are wasting time trying to do their work at the coun¬ cil meetings. Thus, a new approach within sections, possibly to commit the section to long range year-round planning, may be the first break in the activity cycle now supporting the annual meeting concept. To a Botanist, it would mean Academy organized field trips to botanical study areas at the appropriate times of the year. I believe agricultural and industrial facilities could also be visited. Agriculture and environ¬ mental section meetings should be encouraged. I believe these changes are presently possible within the organization of the Academy. A more independent annual meeting committee could operate separately from the council meeting freeing the section chairman to prepare more interdisciplinary programs in¬ stead of waiting long hours at the council meetings to state he needs two rooms and one projector. But a more important idea can be pre¬ sented. I suggest a new role for the Academy Council; the planning and promotion of additional symposia type programs to benefit sections or the entire Academy.

Many members have said they would like senior scientists to pub¬ lish research in our transactions. This would mean a greatly expanded

215

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Transactions Illinois Academy of Science

journal and an increased profes¬ sional value for the transactions. It seems to me we could obtain this goal by first adopting the organized symposium with published papers involving senior scientists in Acad¬ emy activities. I believe we could attract these leaders in the state, region and nation both as partici¬ pants and as readers. In time, I am confident that many would become active in the year-round activities of the Academy and increase the effectiveness of the entire Academy program.

This new role for the Academy, organizing and publishing the re¬ sults of symposia, may make it at¬ tractive to others seeking advice on the social implications of science and technology. By this I mean that some symposia may be helpful to the state and city governmental agencies, especially in developing long range plans for action in sci¬ entific and technological areas to benefit society. The Academy could be an information gathering group, inviting persons from educational, industrial and governmental units to participate. It appears appropri¬ ate to suggest a new affiliation of the Academy with the state govern¬ ment. Precedence for this affiliation already exists. The Director of the State Museum Division of the De¬ partment of Registration and Edu¬ cation of the State of Illinois func¬ tions as Librarian of the Academy, has charge of the distribution, sale, and exchange of publications of the Academy and is a permanent mem¬ ber of the Academy Council.

I have invited the panel members to present their views on the topic of a new role for the Academy, es¬ pecially the possibility of acting as an advisory group to the governor. I will introduce the panel, proceed to their presentations, and invite discussion and comments from the panel and from the floor. Comments

or questions from the floor will be invited after a preliminary round of discussions by the panelists.

The panel consists of Senator Alan J. Dixon, Mr. Milton Thomp¬ son and Dr. Andreas Paloumpis.

Senator Dixon is an attorney, businessman, banker and leader of his party on the floor of the State Senate. In 1950, at 23, he was elec¬ ted to the State House of Represen¬ tatives. He was re-elected to the House for five consecutive terms (1952 to 1960) and served on every major committee. He was Chair¬ man of the House Judiciary Com¬ mittee in 1959 and 1961. In 1962, he was elected to his first term in the Senate and was re-elected in 1966. After only two years in the upper house, he was named Minor¬ ity Whip. He became identified with a particular brand of legisla¬ tion such as the new criminal code; minimum wage bills; judicial and election reform and consumer fraud. Senator Dixon will be our main speaker.

Mr. Milton Thompson received his B.S. degree in Zoology from the University of Minnesota and has been a museum worker since 1937. In 1951, he was appointed Assistant Director of the Illinois State Mu¬ seum, and Director in 1963. In 1952, he was assigned the duty of Librar¬ ian for the Illinois State Academy of Science. In 1968, he was elected President of the Academy. We all appreciate the fine leadership given by Mr. Thompson.

Dr. Andreas Paloumpis received his Ph.D. degree from Iowa State University in 1956 with specializa¬ tion in fisheries research. He was appointed to the Faculty of Bio¬ logical Sciences, Illinois State Uni¬ versity, Normal and, reached the rank of Professor in 1966. In that year, he left his position to become the first President of Winston Churchill Junior College, Pontiac,

Pappelis & Dixon — Role of the Academy

217

Illinois. In July 1969, he accepted the responsibility of Dean of In¬ struction, Illinois Central College, Peoria, Illinois. Dr. Paloumpis has served as Secretary of the Illinois

State Academy of Science from 1962 through 1964.

It is with great pleasure that I present to you the main speaker, Senator Alan Dixon.

A NEW ROLE FOR THE ILLINOIS ACADEMY OF SCIENCE

SENATOR ALAN J. DIXON

Thank you Doctor Pappelis for your kind introduction.

Ladies and Gentlemen of the Academy, I am honored to join you this morning. I must confess I suffered a touch of concern, if not outright foreboding, in this morn¬ ing’s panel discussion concerning a new role for the Illinois Academy of Science. Friends in the academic community have told me that sci¬ entists apply strict standards of academic excellence before permit¬ ting a student to join their ranks as a fully qualified member. You have heard that I am an attorney, legislator and a more or less suc¬ cessful practitioner of the political arts. But since politics and govern¬ ment have not risen to the level of exact sciences, I seriously ques¬ tioned my qualifications for appear¬ ing before you today.

My scientific achievements are not found in any monograph or pe¬ riodical. They can only be found in the notes and reports that passed in a flurry between my high school and college science instructors and parents. I am pained to admit to you that these messages and reports graphically traced a precipitous downward spiral in my scientific aptitudes. I was an early under¬ achiever in the physical sciences.

I have been informed that it is somewhat unprecedented for a member of the Illinois State Senate to address the Academy. I am hon¬ ored to be here to discuss with you areas of mutual concern and inter¬

est which have narrowed the gap between your professions and mine.

After some study and reflection, I am convinced the time is right for an exchange of views between you, as members of our state’s sci¬ entific community, and me, as a representative of our state legisla¬ ture. I hope that some of the com¬ ments I intend to make stimulate you to make further contacts with other legislators and agencies in Springfield. Only in this fashion will we be able to undertake a sci¬ entific dialogue.

I am not qualified to attempt this morning to enter your various fields of research and study. I will limit my remarks to a discussion of some needs that I have begun to perceive clearly during my twenty years in the state legislature. Needs that require your attention and consideration.

Since I was first elected to the general assembly at the age of 23 in 1950, the problems plaguing Il¬ linois, problems that continue to demand some legislative resolution, have multiplied and expanded in geometric fashion. Decisions faced daily by the working legislator have become awesome in complexity. As a Committee Chairman in the House and more recently as Floor Leader of my Party in the Senate, I have participated actively in every ma¬ jor effort to come to grips with the problems that plague our state. During my years in the legislature, the state government in Springfield

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Transactions Illinois Academy of Science

has expanded tremendously. New agencies have been added and old ones expanded to meet continuing demands from the public for im¬ proved and expanded government services. Programs have been initi¬ ated to provide improved health care, better environmental protec¬ tion, more recreational facilities and parks and more effective crime pre¬ vention and control.

But despite the progress that we have made, many of the problems that concerned me 20 years ago as a freshman legislator still plague our state today. And new ones have been added to the catalogue. Urban blight spreads despite our best ef¬ forts to push back its boundaries. Racial harmony and a solution to the nagging problems of urban and rural poverty are not in sight.

Ironically, our successes have pro¬ duced problems that the legislature is required to cope with today. For example, improved public health care has enabled us to reduce Il¬ linois’ infant mortality rate and to lengthen the average life span of residents of the state. Our state’s commercial and industrial develop¬ ment, our welfare standards and our large and flourishing economy have made it a magnet for citizens from poorer states. This enlarged population has created serious drains on our already overburdened resources.

Industrial growth has also been accompanied by the unwanted waste of environmental pollution and the serious problems of auto¬ mation and structural unemploy¬ ment. Industry through automa¬ tion and mechanization has created a vast new requirement to train and retrain thousands upon thou¬ sands of our residents for jobs re¬ quiring higher and more technical skills.

Because of overcrowded condi¬ tions, our public elementary, sec¬

ondary and higher education sys¬ tems are faced with crisis after cri¬ sis. How will we allocate the state’s resources for training new teach¬ ers? What will be the budget for our state’s school systems? Are there new and better programs available to deal with the problems of meeting the educational needs of our adult citizens and taxpayers?

Each year, ladies and gentlemen, the legislature is called upon to deal with this depressing catalogue of nagging problems. The public de¬ mands action, action now, new so¬ lutions, and solutions now, not later. But the legislature is not com¬ posed of experts in these various fields. The legislature is a represen¬ tative group. There are no ecolo¬ gists there to offer their experience and advice on how to come to grips with the problems of environmental waste. Nor will you find highly qualified conservationists able to bring their expertise to bear on bal¬ ancing the state’s need for addi¬ tional recreation space against its need for the preservation of plant and animal life.

The legislature is a representa¬ tive body. Some of its members are labor union men. Others are shopkeepers. There are few doctors. Many are farmers. And most are members of the legal profession. None have a great deal of scientific or technical skill. These ordinary men and women, utilizing their lim¬ ited training in an extraordinary way, are called upon to legislate and to resolve problems that often would require the concentrated con¬ cern of dozens of scientific experts. Make no mistake about it, there is a clear and urgent need for more and better qualified scientific ad¬ vice if the legislature is to cope adequately with the pressing needs of our state.

In order to highlight the legisla¬ ture’s need for an improved system

Pappelis & Dixon — Role of the Academy

219

for obtaining scientific and techni¬ cal advice, allow me to discuss very briefly the present system utilized by the legislature in obtaining this type of information.

In most cases highly motivated and public spirited individuals, who happen to have special qualifica¬ tions to comment on or offer advice concerning a proposed bill, request and are granted time to testify be¬ fore legislative committees. At times committees will invite experts to testify when they are dealing with a particularly thorny issue. The in¬ formation and advice we receive from these meetings often directly influence our views. But even when the testimony is the best available, it is often spotty and incomplete. There is no guarantee that all of the relevant data available on a particular subject has been pre¬ sented. In a typical fifteen minute session of testimony, witnesses sel¬ dom move beyond the broadest generalities and recommendations.

Committee hearings also raise another type of problem in trying to find the best advice available. Many times the testimony offered, even when provided by one with the appropriate academic qualifica¬ tions, represents the views of one or more special interest groups that have a particular stake in the legis¬ lation under consideration. Al¬ though these views have a right to be represented in committee, they rarely are completely reliable. This advocacy system, which at times becomes an adversary system, has been successful up to now. But the time has come when we must sup¬ plement these views with some ad¬ ditional source of scientific advice untainted by a particular special interest and more representative of larger public interests.

Another example of our present information gathering system are the commissions formed by the legis¬

lature from time to time to study a particular major issue. These com¬ missions, composed of legislators and laymen, hire consultants, col¬ lect testimony, and assemble data prior to making recommendations to the Governor or General Assem¬ bly for specific action.

The commission system has been successful in reforming our judicial system. Important recommenda¬ tions have been produced by the State Highway Commission for im¬ proving our state’s system of roads. And more recently, I was a member of the commission which laid the groundwork for calling the consti¬ tutional convention. But the com¬ mission system is not appropriate for gathering and disseminating the amount of scientific information needed by the state government on a continuing basis.

There is one last technique by which the legislature obtains scien¬ tific advice. The executive branch of the state government, which makes legislative recommendations to the General Assembly, has a large staff of trained personnel who serve as advisor to our top policy makers. But these experts are not available directly to the legislature and often represent the views of their agency or political adminis¬ tration.

It has also been my experience that problems tend to outpace their recommendations for solutions. For some reason state agencies often lag behind other institutions such as our universities in projecting so¬ lutions for problems. They some¬ times fail to even recognize let alone respond to a particular pressing problem until it bursts forth in pub¬ lic as a major controversial subject.

As you can see, the legislature’s present system for accumulating technical and scientific advice is clearly unorganized and haphazard.

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Transactions Illinois Academy of Science

It does not provide for a continu¬ ous flow of the most timely data from our state’s scientific commun¬ ity to the legislature. Clearly the individual legislator is not always able to make the kind of educated and informed judgment that is called for in response to a particu¬ lar issue.

If the General Assembly is going to come to grips with the issues of the day that require clear and un¬ biased technical or scientific clarifi¬ cation, the best scientific advice available must be provided for every legislator on a continuing basis. Some agency in the state must un¬ dertake to coordinate and channel a continuous flow of the most re¬ cent scientific information, advice and recommendations to the state legislature. The time has long passed to be satisfied with our present archaic system.

It is clear, ladies and gentlemen, that the Academy is the only state¬ wide organization qualified to co¬ ordinate all of the various scientific interests of the state in undertak¬ ing the task of becoming an official advisory body to the state on scien¬ tific and technical matters. I pro¬ pose for your consideration that the Academy undertake this critically important function.

Since 1907 your organization has written a long and distinguished record of service to the people of the state of Illinois through the promotion of scientific research, and the diffusion of scientific knowledge throughout the state. In undertak¬ ing this new function you would again be undertaking a role of ser¬ vice.

The new advisory function for the Academy could be similar in broad outline if not in specific per¬ formance to that performed by the National Science Foundation for the Federal Government. Although the NSF does serve to channel new

research and ideas to the Federal Government, this is not its sole function. As you also know, it plays a large role in fostering and direc¬ ting pure research activities on a nationwide basis.

Although the Academy may con¬ sider a role of setting statewide sci¬ ence policy similar to that of the National Science Foundation at the national level, I am not qualified to recommend such a course of ac¬ tion. I am concerned, however, with the state government’s need and especially the legislature’s need for more and better scientific advice on a continuing and coordinated scale.

Other proposals have been made to form a statewide Illinois research commission or to create a new of¬ fice of scientific research and tech¬ nology in the Governor’s office. Both of these proposals merit con¬ sideration. The concept of a re¬ search commission is not unprece¬ dented since it is my understand¬ ing that the state of Connecticut is a leader in this area.

Perhaps the Academy could affili¬ ate with such an organization to collect, analyze and summarize sci¬ entific and technological informa¬ tion helpful to all levels and agen¬ cies of state government. I under stand that such an affiliation with the state is not unprecedented since the Academy is presently affiliated and works closely with the Illinois State Museum at Springfield.

In performing its new advisory role, the Academy could consider forming task forces in various prob¬ lem areas to utilize specialists in our universities, industry and gov¬ ernment agencies. Specialists from surrounding states and national and international experts could be called on for counsel and advice when needed.

Both Houses of the legislature, their Committees and the various state agencies in the Executive

Pappelis & Dixon — Role of the Academy

221

Branch, should be permitted to re¬ quest specific studies or informa¬ tion. In return, a grant or stipend would be provided by the state to underwrite the costs of the particu¬ lar study. The work products of the task forces organized by the Academy would be made available to the legislative committee or the agency requesting information or to the appropriate agency when no specific request has been made.

Some topics that might be areas for concentration by early task forces could be agricultural and in¬ dustrial research for statewide eco¬ nomic development. Second, new approaches to our public health needs could be sought. Third, there is a great need for a continual flow of technical information concerning air pollutants, their measurement and control. In time these studies could be expanded to include re¬ search and recommendations from eminently qualified experts in the behavioral sciences.

In conclusion, ladies and gentle¬ men, members of the General As¬

sembly as well as other state offi¬ cials need the advice which may most accurately be produced by the members of this Academy. The need for your skills are great. The prob¬ lems are myriad. New channels of communication must be opened be¬ tween this Academy and the mem¬ bers of the state legislature who must cope with the increasingly complex issues of our time. The job at hand is no small task. It cannot be undertaken without clear plan¬ ning and solid groundwork.

But it is most practical for us, when in this age scientific knowl¬ edge is at a peak, to turn to you to share the value of your wisdom and skills. With your help perhaps we can move on to meet some of the old and new challenges that will face us during the final decades of this century.

I appreciate this opportunity to address you and hope that you will invite me to return to assist you if you choose to undertake this new task.

Manuscript received February 12, 1971

BRYOPHYTES OF GOOSE LAKE PRAIRIE, ILLINOIS

WILLIAM M. ZALES

University of British Columbia, Vancouver, Canada

Abstract. — Thirty species of bryo- phytes are reported new to Grundy Coun¬ ty along with their ecology and a descrip¬ tion of Goose Lake Prairie.

It is fortunate that the State of Illinois has set aside the last re¬ maining fragment of undisturbed prairie for a prairie state park. Ag¬ ricultural development during the early 19th century has led to rapid destruction of Illinois prairie lands, thus little detailed botanical infor¬ mation is available. The scarcity of this type of habitat, that once covered most of Illinois, makes Goose Lake Prairie a precious rem¬ nant of natural history. The follow¬ ing study presents the first list of bryophytes occurring in a natural Illinois prairie.

Goose Lake Prairie is located in Grundy County east of the town of Morris and south of the Illinois River in Sec. 34, T34N, R8E, and Sec. 3, 4, 9 & 10, T33N, R8E, and consists of approximately 1,800 acres. It lies on the valley train of the Old Lake Chicago Outlet which now contains the Illinois River. This area has a basement of Upper Ordovician Richmond Limestone. This is covered by less than 100 feet of Pennsylvania series “Coal Measures” of Carboniferous age; this contains sandstones, shales, clays, limestones, and coals in vary¬ ing succession (Sauer, 1916). This is overlain by a thin surface de¬ posit of Late Wisconsin outwash containing a few minor sand dunes and numerous glacial boulders. The slightly undulating land, reflecting major features of the bedrock sur¬ face, has developed into a luxuriant hydrophytic black soil grassland. Depressions, due to poor stream development, have very wet or sub¬

merged soil but small rises may at times be very dry.

The prairie climate is character¬ ized by sharply contrasting seasons of long summer days with the sun nearly overhead, and short winter days with obliquely shining sun. The frost free season lasts for more than five months with the greatest rainfall in May and June, and the highest temperatures in July and August associated with strong and shifting winds (Sauer, 1916).

The vegetation is characterized by a large number of vascular plant species, 326 phanerogams according to a preliminary list compiled by Swink et al. (1970). These are abun¬ dant in local areas due to aspect dominance of one or several species during some period of the growing season.

Bryophytes do not form a major part of the prairie community. Ac¬ cumulation of dead grass persists year after year and forms a thick dry humus layer; this excludes bryo¬ phytes from moisture and soil. Dense shading by grasses and forbs during favorable growing periods also impedes bryophyte coloniza¬ tion. Bryophytes do not occur as typical luxuriant mats covering ex¬ tensive areas but as small shade- form patches of often attenuated plants. Three notable exceptions are Polytrichum commune and Aulocomnium palustre which have stems long enough to grow through the humus layer, and Sphagnum compactum which produces exten¬ sive hummocks in local areas. Thir¬ teen of the thirty species collected are restricted to more exposed habi¬ tats on dry outwash sand dunes or along edges of stagnant pools. Of nine species collected among dense

222

Zales — Goose Lake Prairie Bryophytes

223

vegetation of wet areas, Amblyste- gium varium, Bryum caespiticium, Leptodictyum riparium, Cephalozia connivens, and Lophocolea hetero- phylla are found later in summer and then only sporadically on sides of grass tussocks. Twenty two per cent of all the prairie species are ephemeral, growing for short peri¬ ods early in spring or in late sum¬ mer, thus avoiding flooding and competition with vascular plants. Fifty per cent were collected with sporophytes during some time of year. Of those collected with ma¬ ture capsules, most were found in exposed habitats and only a few completed their life cycles in com¬ petition with the prairie dominants.

Climax vegetation of hydrophytic prairie soils does not favor most bryophyte species. Those that are present usually occupy the drier elevations or recently disturbed areas, or survive among the vascu¬ lar dominants for short periods of time in spring or late summer.

Bruchia sullivantii and Physco- mitrium pyriforme are endemic to North America; the rest are of wide¬ spread distribution in the northern hemisphere. All species are newly reported for Grundy County. Voucher specimens have been de¬ posited in the bryophyte herbarium of Eastern Illinois University. All collection numbers cited are those of the author. Specimens bearing sporophytes are indicated by an asterisk (*). The nomenclature cited follows Crum et al. (1965) for mosses, and Schuster (1953) for hepaticae. I wish to thank Dr. W. B. Schofield for verifying the speci¬ mens and assistance with this pa¬ per, and the Illinois Nature Pre¬ serves Commission for granting per¬ mission to collect in Goose Lake Prairie.

List of Mosses

Amblystegium varium (Hedw.) Lindb. —

prairie soil under tall grasses, 1017.

Atrichum angustatum (Brid.) B. S. G. — wet prairie soil, 976.

Aulacomnium palustre (Hedw.) Schwaegr. — very common in damper sites, 970*, 1019.

Bruchia sullivantii Aust. — ephemeral on exposed wet mud, 1041*.

Bryum caespiticium Hedw. — on wet sides of grass tussocks, 1036*.

Bryum pseudotriquetrum (Hedw.) Gaertn., Meyer & Scherb. — common in various microhabitats, 965*, 1047.

Ceratodon purpureus (Hedw.) Brid. — dry sandy hills and rocks, 998, 1024*.

Cratoneuron filicinum (Hedw.) Spruce — wet ditches, 1043*.

Desmatodon obtusifolius (Schwaegr.) Schimp. — exposed, wet, sandy soil, 967.

Ditrichum pallidum (Hedw.) Hampe. — - exposed muddy soil, 1042*.

Drepanocladus aduncus (Hedw.) Warnst. — frequent in shallow standing water, 968, 1037. This is the form that resem¬ bles D. fluitans (Hedw.) Warnst. in habit.

Funaria hygrometrica Hedw. — dry cin¬ ders, 1032*.

Leptobryum pyriforme (Hedw.) Wils. — ephemeral on wet mud banks, 1018.

Leptodictyum riparium (Hedw.) Warnst. — frequent on shaded soil, 994, 1046. Leptodictyum is a questionable genus and this specimen will probably turn out to be a form of Amblystegium, “A. riparium fo. Abbreviatum” , see Conard (1959) for a treatment of this problem.

Leptodictyum trichopodium (Schultz) Warnst.— wet soil, 1021*, 975*.

Leskea gracilescens Hedw. — soil at base of Fraxinus, 970*.

Mnium cuspidatum Hedw. — common in various microhabitats, 1045*.

Physcomitrium pyriforme (Hedw.) De Not. — exposed muddy soil, 1023*.

Pohlia nutans (Hedw.) Lindb. — shaded soil, 1025.

Polytrichum commune Hedw. — very com¬ mon in both dry and wet sites, 977*.

Rhynchostegium serrulatum (Hedw.) Jaeg. & Sauerb. — wet prairie soil, 983.

Sphagnum compactum Lam. & DC — form¬ ing large hummocks, 963, 964, 1026.

Weissia controversa Hedw. — ephemeral on exposed clay soil, 1036*.

List of Hepatics

Cephalozia connivens (Dicks.) Spruce — ■ ephemeral on rich humus, 986.

Cephaloziella hampeana (Nees) Schiffn. — shaded soil with Aulacomnium palustre, 984, 1020.

Fossombronia foveolata Lindb. — shaded or exposed soil, 966, 1044.

Lophocolea heterophylla (Schrad.) Dumort. — shaded humus, 978.

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Transactions Illinois Academy of Science

Phaeoceros laevis (L.) Proskauer— shaded or exposed wet mud, 1040*.

Riccia fluitans L.— floating, 1038.

Riccioc.arpus natans (L.) Corda.— floating, 1039.

Literature Cited

Conard, H. S. 1959 Amblystegium. The Bryologist. 62(2) :96-104.

Crum, H., W. C. Steere, L. E. Ander¬ son. 1965. A list of the Mosses of North America. The Bryologist, 68(4) :377- 432.

Sauer, C. O. 1916. Geography of the Up¬ per Illinois Valley. Ill. State Geol. Surv. Bull., 27:1-208.

Schuster, R. M. 1953. Boreal Hepaticae, A Manual of the Liverworts of Minne¬ sota and Adjacent Regions. Amer. Midi. Nat., 49:257-684.

Swink, F., D. Campbell, J. Schwegman, R. Schulenbert, R. Betz. 1970. Plants of the Goose Lake Prairie. 4pp. (mimeo¬ graphed) .

Manuscript received February 1, 1971.

FISHER AND PORCUPINE REMAINS FROM CAVE DEPOSITS IN MISSOURI

PAUL W. PARMALEE

Illinois State Museum, Springfield, Illinois

Abstract. — Remains of the fisher (. Maries pennant!), a mustelid previously unknown from Missouri, are described from four cave and archaeological de¬ posits in the Ozark Highland. A fourth prehistoric locality record for the porcu¬ pine ( Erethizon dorsatum ) in Missouri is presented.

Intensive exploration of Missouri caves, especially those located in the Ozark Highland region, by spe¬ leologists during the past decade has brought to light several signifi¬ cant bone deposits. The time span involved ranges from Late Pleisto¬ cene to present day, and a particu¬ lar cave may contain remains of ex¬

tinct forms such as peccary ( Platy - gonus and Mylohyus), dire wolf (' Canis dims ) and tapir (Tapir) as well as elements of a varied modern fauna. Thus far no completely strat¬ ified deposit has been found and, consequently, animal remains from caves are insufficient to illustrate a clear-cut sequential faunal change through time from the earliest depo¬ sition to the most recent. In spite of the heterogenous nature of such bone deposits — typically the re¬ sult of some form of water action and/or animal activity, the actual presence of particular species often

Figure 1. County map of Missouri showing location of fisher (F) and porcupine (P) finds.

225

226

Transactions Illinois Academy of Science

serves as an indicator of past envi¬ ronmental and species composition change in an area.

Recovery of bones of the fisher (. Martes pennanti) from four cave deposits (Fig. 1) in the Ozark High¬ lands is especially noteworthy. This mustelid presently occurs through¬ out most of the Canadian prov¬ inces and, during early historic times, was locally common in north¬ eastern United States with a range that extended southward in the Ap¬ palachians to North Carolina and Tennessee. Prehistoric archaeologi¬ cal records have shown that its range extended even farther west along the Illinois and Mississippi rivers in Illinois (Parmalee, 1958, 1960a) and south into northwestern Georgia (Parmalee, 1960b) and Ala¬ bama (Barkalow, 1961). Brown (1908) described two isolated teeth, four jaws and a skull section of the fisher that were recovered in the

Pleistocene bone deposit of Conard Fissure in northern Arkansas. This mustelid had not been previously reported from Missouri.

Two of the four records to be de¬ scribed here are based on elements recovered from natural cave depos¬ its, while the other two consist of bones obtained during archaeologi¬ cal investigations in Graham Cave, Montgomery County, in 1966 and Arnold-Research Cave, Callaway County, in 1956. Although all four are termed “cave/’ the latter two represent shallow, overhanging bluff rock shelters once inhabited by peo¬ ples of the Archaic and Woodland cultures. Whether all fishers repre¬ sented in the archaeological depos¬ its were taken by the human inhabi¬ tants or were present as the result of natural causes is, with one ex¬ ception, a matter of speculation.

Arnold-Research Cave, located approximately two miles north of

Figure 2. Fisher elements recovered from Graham (A-C), Brynjulfson (D), Arnold-Research (E) and Bat (F) caves, Missouri.

Parmalee — Missouri Fisher & Porcupine

227

the Missouri River in southeastern Callaway County, was excavated under the direction of J. M. Shippee (Shippee, 1966). A total of approxi¬ mately 36 species of vertebrates (all Recent forms) was identified by Mr. Carl R. Falk; and, in addition to the fisher, three other mustelids were represented (mink, badger, otter). The fisher element consisted of a section of the right maxillary containing P3-4 and M1 (Fig. 2). The animal was a mature adult, the teeth showing moderate wear; judg¬ ing from the large size of the teeth, the individual was probably a male. Graham Cave, situated above Lou- tre Creek approximately 15 miles northeast of Arnold-Research Cave, was excavated under the direction of Dr. Walter E. Klippel; Carl R. Falk and Paul W. Parmalee identi¬ fied the bone remains which in¬ cluded three elements of M. pen- nanti. These consisted of two small, lower right jaws, probably females, both of which contained P2,3,4 and Mi's that showed only slight wear. One of these jaws (Fig. 2, A) exhib¬ ited deep cut marks between the condyle and angular process on the labial surface, plus faint skinning cuts below the alveoli of the canine and Pi, thus indicating utilization of the animal by the Indians. The third fisher element from Graham Cave was a nearly complete right humerus of a large adult, probably a male. There appears to be three individuals represented.

During the summer of 1962, the late Dr. M. G. Mehl and a field crew of students from the Univer¬ sity of Missouri, Columbia, exca¬ vated a horizontal cave shaft lo¬ cated along Bonne Femme Creek, approximately seven miles south- southeast of Columbia, Boone County. The faunal complex of this deposit (Brynjulfson Cave) included remains of extinct forms (peccary, tapir, dire wolf, giant armadillo) as

well as an abundance of Recent species. One fisher element, an in¬ complete lower left jaw containing P2,4 and Mi, was recovered. Based on the large size of this jaw section and the considerable degree of wear on the teeth, it is judged that the animal was an old male (Fig. 2, D).

A large sample of vertebrate re¬ mains was removed from Bat Cave, located five miles northwest of Waynesville, Pulaski County, peri¬ odically from 1961 to 1968; most of the work was carried out under the direction of Mr. J. R. Reynolds, R. L. Foley and Dr. Oscar Hawks- ley. Like Brynjulfson Cave, this cave deposit contained a mixture of remains of modern forms and extinct species such as peccary and dire wolf. Fisher was represented in the faunal complex of Bat Cave by a complete left ulna of a large (male?) adult (Fig. 2, F).

The porcupine ( Erethizon dorsa- tum) is another species, like the fisher, whose range has receded from its known extremities of prehistoric times. In pre-Columbian times this rodent occurred throughout the Ap¬ palachian Mountains south to northern Alabama (Barkalow, 1961), and, in the Midwest, along the Mississippi River in Illinois (Parmalee, 1967) and Missouri (Simpson, 1949). Today, east- of the Mississippi River, the porcupine is not found south of central Wiscon¬ sin, Michigan and Pennsylvania. There have been three published records of the porcupine in Mis¬ souri, all based on elements recov¬ ered in prehistoric cave or archae¬ ological deposits: (1) Two lower jaws from Cherokee Cave (Simp¬ son, 1949), St. Louis County; (2) Sections of a femur, one lower and two upper incisors, and a left jaw from Crankshaft Cave (Parmalee, Oesch and Guilday, 1969), Jefferson County; (3) A lower left jaw section

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Transactions Illinois Academy of Science

from Tick Creek Cave (Parmalee, 1965), Phelps County.

The most recent recovery of por¬ cupine remains in Missouri occurred in the two Brynjulfson Cave depos¬ its. One of these two ‘Twin” caves, located about 100 yards apart along Bonne Femme Creek, Boone Coun¬ ty, has been previously discussed with reference to the recovery of a fisher jaw. The sample of bones from this cave (Brynjulfson Cave No. 1) included complete and/or sections of one skull, five jaws, seven incisors, one cheek tooth, one ulna, five femora and one tibia (Fig. 3, A) of the porcupine. At least four indi¬ viduals, including both adults and juveniles, were represented. An ex¬ tensive faunal sample recovered by Paul W. Parmalee and Ronald D. Oesch periodically during 1968 and 1969 at Brynjulfson Cave No. 2

contained an incomplete humerus, incisor and lower left jaw of the porcupine.

Approximately one-third of the state of Missouri — the central, southern and eastern parts — is classified as Ozark Highland. This region is an old, deeply dissected plateau which now appears as hills with steep intervening valleys cut by numerous streams arising in the higher elevations (Schwartz and Schwartz, 1959). There are many limestone outcrops along the stream banks, and the original mesophytic forest cover was apparently a west¬ ern extension of the oak-hickory climax characteristic of the upland deciduous forests of eastern United States. Juniper and short-leafed pine also occurred throughout this hilly region.

Hagmeier (1956) presents a thor-

Figure 3. Porcupine elements recovered from the Brynjulfson Caves, Boone Co., Missouri.

Parmalee — Missouri Fisher & Porcupine

229

ough discussion of the distribution and habitat requirements of the fisher in North America, and states that . . while they prefer heavy timber, they are frequently seen in open second-growth stands and oc¬ casionally in areas recently burnt over.” Other authorities cited by Hagmeier (ibid.) indicate that fish¬ ers prefer low wet grounds and the banks of streams. This mustelid ap¬ pears quite adaptable within a var¬ ied hilly-wooded-riverine environ¬ ment; the Ozark Highland would have been well suited to its habitat requirements. This hilly, forested region of Missouri would have also provided an ideal habitat for the porcupine, a species that is depen¬ dent upon thick stands of mixed deciduous trees and conifers for food and massive rock outcrops for dens. Since the porcupine is known to be a favorite food of the fisher, there may well have been an inter¬ relationship between the two ani¬ mals, with the range extension and abundance of the fisher dependent to some extent upon the distribu¬ tion and abundance of the porcu¬ pine. In any case, recovery of re¬ mains of these two species in cave and archaeological deposits has es¬ tablished the prehistoric occurrence of fisher and porcupine in the Ozark Highland region of north-central Missouri. Excavation of other de¬ posits will probably bring to light new records of these species, espe¬ cially in regions drained by the Missouri, Osage, Gasconade and Meramec rivers and their tribu¬ taries.

Acknowledgements

I would like to express my appreciation to the following individuals for permission to report on the fisher elements recovered during their field excavations, and for data pertinent to the specimens: Mr. Carl

R. Falk, Midwest Archeological Center, Lincoln, Nebr.; Dr. Oscar Hawksley, Cen¬ tral Missouri State College, Warrensburg, Mo.; and Dr. Walter E. Klippel, Illinois State Museum, Springfield, Ill. Appreci- tion is extended to Charles W. Hodge and Marlin Roos, Illinois State Museum, for photographing the specimens in the Figures.

Literature Cited

Barkalow, Fred. 1961. The porcupine and fisher in Alabama archaeological sites. Jour. Mamm., Vol. 42, No. 4, pp. 544-545.

Brown, Barnum. 1908. The Conard fis¬ sure, a Pleistocene bone deposit in northwestern Arkansas. Memoirs Amer. Mus. Nat. Hist., Vol. IX, Part IV, pp. 157-208; 25 plates.

Hagmeier, Edwin M. 1956. Distribution of martin and fisher in North America. Canadian Field-Naturalist, Vol. 70, No. 4, pp. 149-168.

Parmalee, Paul W. 1958. Evidence of the fisher in central Illinois. Jour. Mamm., Vol. 39, No. 1, p. 153. _ 1960a. Additional fisher rec¬ ords from Illinois. Trans. Ill. State Acad. Sci., Vol. 53, Nos. 1 and 2, pp. 48-49.

_ 1960b. A prehistoric record

of the fisher in Georgia. Jour. Mamm., Vol. 41, No. 3, pp. 409-410.

_ _ .. 1965. Porcupine from the

Ozark Highlands of Missouri. Trans. Ill. State Acad. Sci., Vol. 58, No. 2, pp. 148-149.

_ 1967 A recent cave bone

deposit in Southwestern Illinois. Bull. Nat. Speleol. Soc., Vol. 29, No. 4, pp. 119-147.

Parmalee, Paul W., Ronald D. Oesch, and John E. Guilday. 1969. Pleisto¬ cene and Recent vertebrate faunas from Crankshaft cave, Missouri. Ill. State Mus. Reports of Investigations No. 14, 37 pp.

Schwartz, Charles W., and Elizabeth R. Schwartz. 1959. The wild mam¬ mals of Missouri. Univ. of Mo. Press and Mo. Dept, of Conservation. 341 pp. Shippee, J. M. 1966. The archaeology of Arnold-Research cave, Callaway coun¬ ty, Missouri. Mo. Arch., Vol. 28. 107 pp. Simpson, George G. 1949. A fossil de¬ posit in a cave in St. Louis. Amer. Mus. Novitates, No. 1408. 46 pp.

Manuscript received February 8, 1971

CULTIVATION, LIFE HISTORY AND SALINITY TOLERANCE OF THE TIDEPOOL COPEPOD, TIGRIOPUS CALIFORNICUS BAKER 1912, IN ARTIFICIAL SEA WATER

HARRY W. HUIZINGA

Department of Biological Sciences, Illinois State University , Normal, Illinois 61761

Abstract. — A simple method of culti¬ vating the marine harpacticoid copepod, Tigriopus Calif ornicus Baker 1912, using artifically prepared sea water enriched with Purine laboratory rat chow is de¬ scribed. The copepod feeds upon a mixed diet of unicellular algae, protozoa, bac¬ teria and organic matter. The life history from egg through nauplius, copepodite and reproductive adult takes 18 to 21 days at 23 C. Life history stages are illus¬ trated. The copepod is tolerant to a wide range of salinity (21.2 to 75.3 o/oo) and shows an optimum growth and reproduc¬ tion in a salinity of 42.3 to 47.00 o/oo. Potential uses of this hardy and easily cultured organism in research and teach¬ ing are discussed.

The harpacticoid copepod, Tigri¬ opus californicus Baker 1912, may be of interest to the invertebrate zoologist who is looking for a ver¬ satile marine organism to use in re¬ search and teaching. T. californicus lives in splash pools above the high tide zone along the rocky California coastline. It is a hardy organism that is able to withstand the wide fluctuations of temperature and sa¬ linity that are found in the un¬ stable tidepool habitat (Monk, 1941).

The copepod is little-known, but potentially a valuable animal for use in various kinds of research. Tigriopus spp. have been used in ecological studies (Fraser, 1936), nutritional study (Provasoli, et ah, 1959; Gilat, 1967), genetic study (Vacquier, 1962), radioactive tracer uptake (Chipman, 1958; Lear and Oppenheimer, 1962), as a food for marine killifish (Faye, personal com¬ munication) and as an experimental intermediate host for nematode par¬ asites (Huizinga, 1966).

Most marine copepods are diffi¬ cult to cultivate since they have exacting physiological requirements and must be fed a variety of spe¬ cially cultured algae. However, T. californicus is a benthic filter-feeder and its nutritional needs are easily satisfied in laboratory culture with a diet of mixed unicellular organ¬ isms.

This paper describes a simple method for raising T. californicus in artificially prepared sea water culture. Observations are given on the life history, reproduction and salinity tolerance.

Materials and Methods

Cultivation techniques. T. califor¬ nicus can be obtained from Dr. Rimon Fay, Pacific Bio-Marine Supply Company, Box 536, Venice, California 90291. The animal is easily shipped via air mail in sealed polyethylene bags containing about one liter of sea water with an over¬ laying air space. Field collected copepods are placed into a standard 8 inch diameter finger bowl and left in the natural sea water for about one week to allow for equilibration to the laboratory conditions. They are fed Purina laboratory rat chow which is finely ground with a blend¬ er. A small amount of food (150 mg) is added at 4 to 7 day intervals to a bowl containing 1300 ml of sea water and approximately 500 adult copepods. The culture is covered and maintained at room tempera¬ ture (20-25 C), without aeration. The initial water level is marked on the side of the bowl with a wax

230

Huizinga — Cultivation of Tigriopus

231

3 4

Plate 1. Life history stages of the harpacticoid copepod, Tigriopus calif ornicus. Figure 1. Male (left) and female in copulation, magnification 58 X original. Figure 2. Antennae of male, modified as clasping organs (c), 174 X.

Figure 3. Copepodite, 112 X.

Figure 4. Nauplius, 194 X.

Figure 5. Female with attached egg sac (E), 57 X.

Figure 6. Larval stage of the nematode Contracaecum spiculigerum (arrow) with¬ in the hemocoel of T. calif ornicus, 220 X.

232

Transactions Illinois Academy of Science

pencil, and water lost to evapora¬ tion is replaced with distilled water.

After the original culture begins to show multiplication of nauplii and copepodite stages (Figs. 3 and 4), subcultures are made at appro¬ ximately two to three week inter¬ vals, depending upon the room tem¬ perature and growth characteristics of the culture. The frequency of subculturing is not critical, since a neglected culture will remain viable for several months with only a few copepods present. The animal can withstand extremely crowded con¬ ditions, but will attain a maximum population density in about 24 days, beyond which a decline of numbers will occur.

Subculturing is done by stirring the bottom debris and copepods into suspension with an artist’s paintbrush and pouring one-half of the contents into a second bowl. Each bowl is then filled to the 1300 ml mark with artificially prepared sea water (42.6 gms of synthetic sea salts per liter of deionized tap water, salinity 35 parts per thou¬ sand, specific gravity 1.025, Dayno Sea Salts, Supreme Products, Lynn, Mass.). The resulting mixture is 50 parts original sea water and 50 parts artificial sea water. Trace elements were not added. Part of the original bottom debris is included in the subculture, since it contains the needed food organisms from the original field-collected sea water shipment (e.g. unicellular algae, bac¬ teria and protozoa). The copepods will not thrive if they are placed into unconditioned artificially pre¬ pared sea water. A critical factor in the growth of Tigriopus is the quantity of microorganisms present. In my experience, a slightly cloudy medium indicates an optimum growth of microorganisms which are feeding upon the powdered food. If the culture becomes transparent and low in microorganisms, an im¬

mediate decline in the numbers of copepods will be observed. Over¬ feeding is not critical, since Tigri¬ opus is tolerant of considerable pu- trifaction. The organism does equally well with or without aera¬ tion.

Adult copepods are harvested from cultures with a sieve that has a mesh of 500 microns. This allows the passage of smaller nauplii and copepodite stages and retains the larger adults which are transferred to other containers.

Attempts to raise T. calif ornicus in a constant temperature plant in¬ cubator (25 C) with high intensity lighting (100-200 foot candles, 13 hours duration) failed. This light intensity stimulated excessive growth of undesirable red filamen¬ tous algae which caused an imbal¬ ance of the system and death of copepods. The fluorescent lighting of my office (20-40 foot candles, 8-10 hours duration) allowed for adequate growth of algae and pro¬ duced good copepod cultures. A sin¬ gle-celled green alga ( Chlorococcum sp.) was the predominant food item found in the intestinal contents of copepods along with lesser amounts of a blue-green alga ( Oscillatoria sp.) bacteria, protozoa ( Oxyrrhis marina, Euplotes spp.) and non-living par¬ ticulate organic matter.

When a white fungus was ob¬ served to grow upon the bottom debris, subcultures were made and the fungal contaminant discarded. Tigriopus was resistant to infection, but the toxic effect of the fungus appeared to limit the growth of or¬ ganisms serving as food for the copepod. Fungal contamination was minimized by autoclaving the pow¬ dered food and storing in a dry con¬ tainer.

The following method was used to study the life history of T. cali- fornicus. A gravid female (Fig. 5) was transferred from the stock cul-

Huizinga — Cultivation of Tigriopus

233

ture with a capillary pipette into a 4 inch diameter petri dish contain¬ ing 40 ml of filtered and conditioned sea water. Ten milligrams of pow¬ dered food (see above) was added. The dish was placed in a lighted room at a temperature of 23 C. Daily observations were made using

a dissection microscope with bright illumination and a black back¬ ground. A few drops of stock Harris hematoxylin stain, added to the dish at the termination of an ex¬ periment, caused copepods to ad¬ here to organic matter on the bot¬ tom of the dish. The stained cope-

Table I. — Progeny from single egg sacs of isolated female Tigriopus californicus.

Isolated

Day

Numbers of Progeny

Adults

Female*

Killed

Nauplii

Copepodites

Male

Female

1

2

9

0

0

0

2

2

13

0

0

0

3

2

15

0

0

0

4

5

2

13

0

0

5

5

5

10

0

0

6

6

0

12

0

0

7

8

0

13

0

0

8

12

0

19

0

0

9

17

0

0

9

3

10

17

0

0

13

8

*Females were removed after one egg sac

was laid.

Table II. — Salinity tolerance of Tigriopus californicus in artificial sea water.

Concentration Maximum Reproduction Overall

Sea Salt, Salinity Survival of Nauplii Success

Gms per liter o/oo in Days & Copepodites Behavior of Culture

126.0

114.6

103.1

91.6 80.9

68.7

57.3

51.6

45.8

42.6

40.1

34.4

28.6

25.8

22.9

20.0

17.2

14.3

11.4 8.8 5.7 2.9 0.0

103.5

1

94.1

7

84.7

10

75.3

40*

65.9

40*

56.4

40*

47.0

40*

42.3

40*

37.6

40*

35.0

40*

32.9

40*

28.2

40*

23.5

40*

21.2

40*

18.8

16

16.5

13

14.1

11

11.7

11

9.4

5

7.0

4

4.7

4

2.3

3

0.0

2

+ + + + + + + + + + + + + +

+

+

+

A,E**

A,E

A,E,M

A,E,M

M,N

M,N

M,N

M,N

M,N

M,N

M,N

M,N

M,N

M,N

E,M,N

A,E,M

A,E,M

E,M

A,E

A,E,M

A,E

A,E

A,E

4

4

4

3

3

2

1

1

1

1

1

2

2

2

3

4 4 4 4 4 4 4 4

* * *

terminated

**A = abnormal movement and feeding, E = epiphytes, M = mating, N = normal movement and feeding

***1. optimum, maximum numbers and survival, 2. average, moderate numbers and survival, 3. below average, limited reproduction and survival, 4. unsuccessful, poor to no reproduction, premature death.

234

Transactions Illinois Academy of Science

pods continued to move in place for several minutes before death and were easily counted using a dissecting microscope and ruled pe- tri dish.

Copepods were placed into vari¬ ous concentrations of artificial sea water to test their salinity toler¬ ance (Table II). A stock concentra¬ tion of sea water was prepared by adding 126 gms of synthetic sea salt per liter of deionized tap water and heating to boiling. Successive 10 per cent dilutions of the stock con¬ centration were made with deion¬ ized water. Since the calcium sul¬ fate, calcium manganate and cal¬ cium carbonate salts tended to pre¬ cipitate from the supersaturated so¬ lution, they were stirred into uni¬ form suspension before dilutions were made. The salinity and num¬ ber of grams of sea salt per liter for each dilution were calculated based upon the manufacturer’s specifica¬ tions for oceanic sea water (Table II, Dayno Sea Salt, 42.6 gms per liter equals 35 o/oo).

Results

Life History. After the female copepod drops an egg sac, nauplii begin to hatch from the eggs within 24 hours (Fig. 4). These move along the bottom of the dish as they feed upon microorganisms. The first nau- plius molts three times giving rise to three additional nauplii stages. The animal increases slightly in size after each molt. After 5 or 6 days, the final nauplius stage de¬ velops into the copepodite which resembles the miniature adult (Fig. 3). There is also a sequence of four copepodite stages over a period of 8 to 9 days. The final copepodite then develops into the adult cope- pod (Fig. 1). Development from the egg to adult takes about 15 to 18 days at 23 C.

Young male and female copepods begin to mate immediately. There

is a striking sexual dimorphism and the male has specially modified an¬ tennae which are used to clasp the female during copulation (Figs. 1 and 2). The male pursues the fe¬ male vigorously from behind and grasps her cephalothorax with a quick motion. He then rides the female for several minutes to hours during which the sperm are depos¬ ited in the seminal receptacle. The female goes about her usual feeding behavior, apparently unaffected by the male in tow. An inseminated female will begin to produce an egg sac in about 3 to 4 days. She will continue to produce egg sacs, in iso¬ lation from the male, by fertilizing the eggs with stored sperm from a single insemination. New egg sacs appear every 2 to 3 days. Isolated females were observed to lay three consecutive egg sacs. The average number of eggs per sac is 18. At room temperature, the entire life cycle from egg to egg takes 18 to 21 days. The numbers of progeny pro¬ duced by isolated gravid female copepods (Fig. 5) at various time intervals are given in Table I.

Salinity Tolerance. T. calif ornicus survived and reproduced for over 40 days in a wide range of salinity (21.2 to 75.3 o/oo, Table II). Above and below this range, the feeding, reproductive behavior and survival were abnormal. Epiphytic fungi grew upon the exoskeleton of ab¬ normally sluggish copepods.

Copepods remained alive for one day in 84.7 o/oo which was nearly three times the concentration of normal sea water (35 o/oo), but they were sluggish and twitched convulsively. The copepod survived for two days in deionized water.

Moderate growth and reproduc¬ tion of copepods was observed in the concentrations of 21.2 and 56.4 o/oo. Abundant cultures developed in the range of 32.9 to 47.0 o/oo. Based upon maximum reproduction

Huizinga — Cultivation of Tigriopus

235

of nauplii and copepodites plus vig¬ orous feeding and mating behavior, the optimum sea salt concentration was 42.3 to 47.0 o/oo.

Discussion

The simple method described here has been used in my laboratory for the continuous cultivation of T. calif or nicus for over five years. The copepod is hardy and offers con¬ siderable potential for use in re¬ search and teaching, such as: an assay organism for environmental pollutants (Mileikovsky, 1970), model of population dynamics, study of nutritional requirements, and copepod reproductive behavior.

T. californicus is susceptible to experimental infection with Con- tracaecum, a larval nematode para¬ site (Huizinga, 1966, and Fig. 6 of this paper). The copepod is also capable of withstanding consider¬ able salinity variation. It may there¬ fore serve as an experimental inter¬ mediate host for various larval hel¬ minth parasites found in marine or estuarine environments.

The tolerance of the copepod to a wide range of salinity is not sur¬ prising in view of its natural habi¬ tat which is subject to evaporation and periodic dilution by rainwater. Tigriopus is also tolerant to consid¬ erable putrifaction and this is prob¬ ably an adaptation to the tidepool which is normally exposed to fecal enrichment from marine birds.

T. californicus was observed to feed upon a mixed diet of unicel¬ lular algae, protozoa, bacteria and organic matter. Provasoli et al. (1959) and Gilat (1967) showed sim¬ ilarly that T. brevicornis and T. californicus require a diet of mixed algae and bacteria for successful reproduction in laboratory culture.

Since T. californicus is easily cul¬ tivated, it can be used conveniently in the classroom to demonstrate the life history and reproduction of

a marine copepod. The following student exercise is suggested to com¬ pare the theoretical and observed reproductive potential. Since a fe¬ male copepod produces on average of 18 eggs per sac, and assuming a 50-50 sex ratio, the FI generation will contain about nine females. Upon maturity in 21 days, each FI female will then produce at least one egg sac giving rise to a total of about 162 F2 progeny. Continuing with a similar theoretical line of mathematics and optimum growth conditions, the F3 generation will contain about 1458 progeny in about 60 days. The actual number of off¬ spring will be several times the above calculated number, since a single female copepod usually lays several consecutive egg sacs. Mor¬ tality must also be taken into ac¬ count. The student can perform total or sample counts of the cope¬ pod stages present in the culture over a several week period (as Ta¬ ble I, Results). This study can be used to illustrate the high rate of biological productivity shown by the primary consumer copepod which serves as an important food organism for marine fish.

Summary

1. Tigriopous californicus was cul¬ tured in a simple system using artificially prepared sea water enriched with powdered Purine laboratory rat chow. The cope¬ pod fed upon a mixed diet of uni¬ cellular algae, protozoa, bacteria and organic matter.

2. The life history of T. californicus from egg through four nauplii, four copepodites and the repro¬ ductive adult took 18 to 21 days at 23 C.

3. The copepod survived and re¬ produced in a wide range of sa¬ linities (21.2 to 75.3 o/oo). A sa¬ linity range of 42.3 to 47.0 o/oo

236

Transactions Illinois Academy of Science

was optimum for growth and reproduction.

4. Potential uses of this hardy and adaptable organism in research and teaching are discussed.

Acknowledgements

This study was supported in part by research grant No. 69-21 from the Illinois State University foundation. I wish to thank Mrs. Faye Townsend for technical help and Dr. Sung Y. Feng of the Marine Research Laboratory, University of Con¬ necticut, for reviewing the manuscript.

Literature Cited

Chipman, W. A. 1958. Accumulation of radioactive materials by fishery organ¬ isms. Proc. Gulf Caribb. Fish. Inst. 11th Annual Session 97-100.

Faye, W. E. (personal communication) University of North Carolina, Institute of Fisheries Research, Moorehead City, N. C.

Fraser, J. H. 1936. The occurrence, ecology and life history of Tigriopus fulvus (Fischer). J. Mar. Biol. Assoc. U. K. 20:532-536.

Gilat, E. 1967. On the feeding of a ben-

thonic copepod, Tigriopus brevicornis. Sea Fish Res. Sta. Bull., Haifa, Isr. 45:79-94.

Huizinga, H. W. 1966. Studies on the life cycle and development of Contra- caecum spiculigerum (Ascaroidea: He- terocheididae) from marine piscivorous birds. J. Elisha Mitchell Sci. Soc. 82: 181-195.

Lear, D. W. and Oppenheimer, C. H. 1962. Consumption of microorganisms by the copepod, Tigriopus calif ornicus . Limn, and Oceanogr. Suppl. 7:63-65.

Mileikovsky, S. A. 1970. The influence of pollution on pelagic larvae of bottom invertebrates in marine nearshore and estuarine waters. Mar. Biol. 6:350-356.

Monk, C. R. 1941. Marine harpacticoid copepods from California. Trans. Amer. Micr. Soc. 60:75-103.

Provasoli, L., Shiraishi, K., and Lance, J. R. 1959. Nutritional idiosyncrasies of Artemia and Tigriopus in monoxenic culture. Ann. New York Acad. Sci. 77:250-261.

Vacquier, V. 1962. Hydrostatic pressure has a selective effect on the copepod Tigriopus. Science 135:724-725.

Manuscript received March 12, 1971

INCIDENCE OF MERCURY IN ILLINOIS PHEASANTS

WILLIAM L. ANDERSON AND PEGGY L. STEWART

Illinois Natural History Survey, Urbana, 61801, and Stewart Laboratories, Inc., Knoxville, Tennessee, 37921

Abstract. — Selected tissues from 20 pheasants collected in east-central Illinois during August 1970 were analyzed for elemental mercury by emission spectro- graphy. Frequencies of occurrence and mean concentrations were found to be 35 per cent and <0.06 ppm in kidneys, 40 per cent and 0.03 + 0.01 ppm in livers, 15 per cent and 0.32 ± 0.30 ppm in brains, 25 per cent and 0.02 ± 0.01 ppm in leg muscles, 15 per cent and 0.03 ± 0.02 ppm in sternal muscles. The high mean con¬ centration in brains was due to 5.93 ppm found in one bird. The second highest in¬ dividual concentration was 0.44 ppm and occurred in kidneys. Eight soil samples, collected from the fields in which the pheasants were taken, contained a mean of 0.02 + 0.01 ppm of mercury. Thus, neither pheasants nor soils in east-central Illinois appear to be contaminated with potentially dangerous levels of mercury.

Mercury contamination became a subject of considerable concern in 1969 and 1970. Ring-necked pheas¬ ants ( Phasianus colchicus) and Hun¬ garian partridges ( Perdix perdix) in Alberta, Canada, and in Montana were found to contain potentially dangerous levels of mercury (Wish- art 1970, Dunkle 1969). Relatively high levels of mercury were also found in several species of North American waterfowl (National Wildlife Federation 1970), and mer¬ cury contamination prompted state governments to impose restrictions on fishing in many lakes and rivers (Sport Fishing Institute 1970). The pheasants and partridges in Alberta and Montana apparently became contaminated by eating seed grain treated with organic mercury fun¬ gicides.

The purpose of this study was to determine the incidence of mercury in selected tissues and organs of pheasants in east-central Illinois, the state’s better pheasant range. This portion of Illinois is under in¬

tensive cultivation, with about 70 per cent of the total land area planted to corn and soybeans an¬ nually (calculated from Illinois Co¬ operative Crop Reporting Service 1970:64). According to M. C. Shurt- leff. Professor of Plant Pathology, University of Illinois, Urbana (per¬ sonal communication), mercury fun¬ gicides were used sparingly in Illi¬ nois to treat seed grain of small grains — wheat, oats, barley, and rye — through 1969. However, all recommendations for uses of mer¬ cury compounds in Illinois agricul¬ ture were discontinued in 1970. About 4 per cent of the total land area in east-central Illinois was planted to small grains in 1969 (cal¬ culated from Illinois Cooperative Crop Reporting Service 1970:68, 72).

Methods

The pheasants used for this study were captured by nightlighting (Labisky 1968) during the nights of 24 and 25 August 1970. Ten of the pheasants were taken from four fields (three wheat stubble and one idle) located in southwestern Cham¬ paign County (townships T17N, R8E and T18N, R8E) and 10 were taken from four fields (one oat stub¬ ble, two brome, and one idle) loca¬ ted in southeastern Livingston County (townships T25N, R7E and T25N, R8E). Of the 20 pheasants collected, five were juvenile hens, five were juvenile cocks, nine were adult hens, and one was an adult cock. The juveniles were aged to the nearest week by examining ad¬ vancement of molt of the primary flight feathers (Etter et al. 1970).

The pheasants were held over-

237

238

Transactions Illinois Academy of Science

night in wooden crates, and were then weighed, decapitated, and dis¬ sected, one at a time, between 7 :30 AM and 11:00 AM. The following tissue samples were excised from each bird and saved for analysis: both kidneys (3. 2-5.6 g), entire liver (9.0-15.6 g), entire brain (1.9-3. 8 g), 10-20 g of leg muscle (primarily the femoris), and 16-49 g of sternal mus¬ cle (pectoralis thoracica ). The tissue samples were freed of extraneous material, weighed, and packaged in polyethylene bags or vials. The packaged samples were placed on dry ice within 15-20 minutes after the birds had been killed. Metal instruments (scissors, forceps, and scalpels) were used to dissect the pheasants.

A sample of soil was collected from each of the eight fields in which the pheasants were captured, and was saved for analysis. Each sample was obtained by collecting three subsamples at approximately the same location in the field — i.e., collected within 5 m of each other — and composited. The samples were taken from the upper 3 cm of soil. They were stored in polyethy¬ lene bags until they could be ana¬ lyzed.

Analyses of the tissue samples and the soil samples for mercury were performed at Stewart Labora¬ tories, Inc., Knoxville, Tennessee. After removing water from the tis¬ sues by freeze-drying (at < 1 mm Hg), the organic matter was de¬ stroyed in a manner that did not re¬ sult in the loss of mercury. A num¬ ber of investigators (Schoniger 1955 and 1956, Southworth et al. 1958, Gutenmann and Lisk 1960) have shown that the Schoniger method of combustion is applicable to the determination of mercury in organic compounds. By the use of this method, organic matter is destroyed in the combustion and the volatile elements are retained in a closed

system. The combustion products are absorbed in a reagent (added to the system prior to combustion), which serves to fix the mercury and convert it to a form suitable for determination. Appropriate modifications were made to increase the specificity of these preparations for determination of mercury in nearly all organic matter. The soil samples were pretreated by a con¬ ventional dissolution procedure that resulted in a solution similar to that of the organic samples.

The mercury present in solution was withdrawn by an extractant that contained as its major com¬ ponent a highly volatile organic. By a process of successive evapora¬ tions (no heat), a concentrated so¬ lution was evaporated onto a buffer (carrier) compound packed in a car¬ bon electrode. The carrier acts sim¬ ilar to a distilling column and al¬ lows for quick introduction of all the mercury from the sample into an electric arc. The light emission from the D.C. arc excitation was recorded by an emission spectro¬ graph.

Although the absolute sensitivity of the spectrographic technique usu¬ ally is not low for mercury, the ability to burn the extract of sam¬ ples > 1 g in size in a single elec¬ trode results in adequately low de¬ tection limits for the technique. The lower limits of detection were 0.02 fig per g of wet liver, leg muscle, and sternal muscle; 0.06 jag per g of wet kidneys and brain; and 0.01 fig per g of dry soil. The absolute concentration ranges of the spec- trochemical method employed were 0.01-10.0 fig of mercury. Spike re¬ coveries averaged 82 per cent in the 0.01-0.10ja g range, 88 per cent in the 0. 10-1. Oja g range, and 95 per cent in the 1.0-10.0 jag range.

Concentrations of mercury in the pheasant tissues and in the soil sam¬ ples are presented in ppm (jag per

Anderson & Stewart — Mercury in Pheasants

239

g). When mean concentrations were calculated, values less than the low¬ er limits of detection were consid¬ ered to be 1/10 the lower limit.

Findings and Discussion

The pheasants from Champaign County and from Livingston Coun¬ ty did not differ appreciably in body weight, advancement of molt of the primary flight feathers, or incidence of mercury in their bodies, when the possible influence of sex and age was taken into consideration. Thus, data for birds from the two coun¬ ties were combined for statistical analysis and presentation.

Mean weights of the pheasants were 486 ± 40 g for juvenile hens, 719 ± 68 g for juvenile cocks, and 813 ± 12 g for adult hens. The adult cock weighed 1,043 g. Mean ages of the juvenile birds were 9.4 ± 0.4 weeks for hens and 10.6 ± 0.7 weeks for cocks. The adult hens had molted an average of 6.6 ± 0.3 primaries; the adult cock had molted 7 pri¬ maries. These physical characteris¬ tics are considered to be typical of pheasants in east-central Illinois during late August.

The frequencies of occurrence and the concentrations of elemental mer¬

cury in the pheasant tissues are summarized in Table 1. From the data in the table it appears that the incidence of mercury in juvenile birds and in adult birds was similar. When all 20 pheasants were con¬ sidered, mercury was detected in 35 per cent of the kidneys, 40 per cent of the livers, 15 per cent of the brains, 25 per cent of the leg mus¬ cles, and 15 per cent of the sternal muscles. However, mercury was de¬ tected in at least one of the five tis¬ sues in 85 per cent of the pheasants included in this study.

Mean concentrations of mercury were low (0.07 ppm or less) in all tissue samples except brains of the juvenile pheasants. The high mean concentration in brains is attribut¬ able to a high value (5.93 ppm) found in one juvenile. This high value appears to be analytically correct. However, it is something of a paradox in that it occurred in a bird in which the other tissues had low values (0.08 ppm or less). We offer no explanation as to why mercury was abundant in the brain of this bird, except to point out that one of us (PLS) has observed similar phenomena with mercury in other biological materials. An

Table 1. — Incidence of mercury in pheasants in east-central Illinois, August 1970.

Frequency

of

Occurrence

Concentrations (ppm)* Mean Median

Highest

Kidneys

Juveniles

3/10

<0.06

<0.06

0.12

Adults

4/10

0.07+ 0.04

<0.06

0.44

Liver

Juveniles

3/10

<0.02

<0.02

0.11

Adults

5/10

0.04+ 0.02

<0.02

0.20

Brain

Juveniles

2/10

0.61 + 0.59

<0.06

5.93

Adults

1/10

<0.06

<0.06

0.28

Leg Muscle

Juveniles

3/10

0.02+ 0.01

<0.02

0.10

Adults

2/10

<0.02

<0.02

0.08

Sternal Muscle

Juveniles

1/10

<0.02

<0.02

0.08

Adults

2/10

0.05+ 0.04

<0.02

0.40

*/JLg per g of wet weight.

240

Transactions Illinois Academy of Science

occasional high value appears to be the rule rather than the exception.

Because humans normally eat the muscular portions of pheasants, the incidences of mercury in leg muscle and in sternal muscle are of particular interest. Concentrations in the five samples of leg muscle that contained mercury above the limit of detection were 0.02, 0.06, 0.08, 0.08, and 0.10 ppm. Similarly, concentrations in the three sam¬ ples of sternal muscle that con¬ tained detectable levels were 0.08, 0.10, and 0.40 ppm. Except for sternal muscles of adult birds, mean concentrations in muscular tissues were 0.02 ppm or less. The rela¬ tively high mean concentration in sternal muscle of adults (0.05 ppm) was due to 0.40 ppm that was found in one bird. For comparison, an average of 0.18 ± 0.03 ppm of mer¬ cury was found in flesh of pheas¬ ants and partridges collected in Montana (presumably in 1969) ; the frequency of occurrence was 100 per cent in a sample of 20 birds (Dunkle 1969:3). In Alberta, con¬ centrations of mercury in muscular tissue of these game birds, collected in early summer 1969, averaged 0.45 ppm (Wishart 1970:5).

The U.S. Food and Drug Admin¬ istration has established 0.50 ppm as a temporary guideline for mer¬ cury in fish, but the agency has not set tolerance levels for game birds, poultry, or other human foods. How¬ ever, in Canada, the Food and Drug Directorate recently adopted 0.50 ppm as a temporary tolerance level for mercury in game birds. The World Health Organization suggests 0.05 ppm as the tolerance level in human foodstuffs. If 0.50 ppm is considered permissible for mercury levels in pheasants, all 20 of the birds examined during the present study would be safe for human con¬ sumption. On the other hand, if the permissible level is considered to be

0.05 ppm, four samples (20 per cent) of leg muscle and three sam¬ ples (15 per cent) of sternal muscle might be considered unfit for hu¬ man food. Mean concentrations of mercury in both leg muscle and sternal muscle of both juvenile pheasants and adult pheasants did not exceed 0.05 ppm (Table 1).

The samples of soil collected in the fields in which the pheasants were captured contained an average of 0.02 ± 0.01 ppm of mercury when expressed on a dry-weight basis. Mercury was detected in four of the eight samples, the highest individual concentration being 0.05 ppm. Because mercury occurs nat¬ urally in the environment (Bowen 1966:187), small amounts of this heavy metal might be expected to be present, in the soil samples, as well as in the pheasants.

When interpreted in light of ex¬ isting tolerance levels and avail¬ able geochemical information, find¬ ings of this study indicate that neither pheasants nor soils in east- central Illinois are contaminated with potentially dangerous levels of mercury.

Acknowledgments

Thanks are expressed to the following personnel of the Illinois Natural History Survey: To Glen C. Sanderson for tech¬ nical advice, to William R. Edwards and Stanley L. Etter for reading the manu¬ script, to Helen C. Schultz for editorial assistance, and to James W. Seets for help in collecting pheasants. Anna M. Yoa¬ kum, Stewart Laboratories, Inc., assisted with the analytical work and reviewed the manuscript.

Literature Cited

Bowen, H. J. M. 1966. Trace elements in biochemistry. Academic Press, London and New York. 241 pp.

Dunkle, F. H. 1969. Report on the “hot” pheasants. Montana Outdoors 4(11) :l-3.

Etter, S. L., J. E. Warnock, and G. B. Joselyn. 1970. Modified wing molt criteria for estimating the ages of wild juvenile pheasants. J. Wildl. Mgmt. 34(3) :620-626.

Anderson & Stewart — Mercury in Pheasants

241

Gutenmann, W. H., and D. J. Lisk. 1960. Rapid determination of mercury in apples by modified Schoniger com¬ bustion. J. Agr. and Food Chem. 8(4): 306-308.

Illinois Cooperative Crop Reporting Service. 1970. Illinois agricultural sta¬ tistics. Annual summary 1970. Illinois Department of Agriculture and U.S. Department of Agriculture, Spring- field, Illinois. Bulletin 70-1. 99 pp. Labisky, R. F. 1968. Nightlighting: its use in capturing pheasants, prairie chickens, bobwhites, and cottontails. Illinois Natural History Survey Biol. Notes 62. 12 pp.

National Wildlife Federation. 1970. More mercury in waterfowl. Conserva¬ tion News 35(17) :5-6.

Schoniger, W. 1955. Eine mikroanaly-

tische Schnellbestimmung von Halogen in organischen Substanzen. Mikrochim. Acta 1:123-129.

_ 1956. Die mikroanalytische

Schnellbestimmung von Halogenen und Schwefel in organischen Verbindungen. Mikrochim. Acta 4-6:869-876.

SOUTHWORTH, B. C., J. H. HODECKER, and K. D. Fleischer. 1958. Determi¬ nation of mercury in organic com¬ pounds: a micro and semimicromethod. Anal. Chem. 30(6) :1152-1153.

Sport Fishing Institute. 1970. Fishing restrictions re mercury. SFI Bulletin 219:2-3.

Wishart, W. 1970. A mercury problem in Alberta’s game birds. Alberta Lands — Forests — Parks — Wildlife 13(2) :4-9.

Manuscript received March 5, 1971

LONGITUDINAL PATTERN OF NUCLEAR SIZE IN BULB SCALE EPIDERMIS OF ALLIUM CEPA AND CHANGES IN SIZE IN RESPONSE TO NECKROT

F. B. KULFINSKI AND A. J. PAPPELIS

Department of Biological Sciences, Southern Illinois University, Edwardsville 62025, and Department of Botany, Southern Illinois University, Carbondale 62901.

Abstract. — Nuclear size in onion bulb scale inner epidermis cells was found to vary, increasing in size from base toward the equator and decreasing in size toward the apex of the bulb. The mean nuclear area in basal cells was 3.4 x 10-6 cnu. The nuclear area increased to a maximum of 9.1 x 10-6 cm2 near the equator and de¬ creased to 3.6 x 10-6 cm2 near the apex. Nuclear size may be related to meriste- matic activity near basal cells, to cell en¬ largement patterns during bulb develop¬ ment, and to cellular aging and death at the apex following harvest.

Epidermal nuclei adjacent to mycelium of neckrot pathogens responded to infec¬ tion by decreasing in size. Such nuclei averaged 47 to 81 per cent of normal in area (X.S.), the amount of reduction depending apparently on their initial size.

Nuclear diameter and nuclear dry mass were studied in outer epider¬ mis cells of the equatorial areas of the inner to outermost bulb scales of onion by Courtis (1964). He re¬ ported that average cell area, nu¬ clear diameter, and nuclear dry mass increased from inner to outer bulb scales and that these characteris¬ tics were closely correlated. These increases were suggested to be age related since outer leaf bases are older than inner ones.

Changes in nuclear size have been reported in a number of studies, a nucleus frequently increasing and then decreasing in response to the same factor. Increases and decreases were reported during cell death by Ogilvie (1962), Cameron (1951), and Bessis (1964), in irradiated cells by Eckert and Cooper (1937), during autolysis by Hasten (1958), and in response to fungus infection by Goodman, et al. (1967). Increases have been reported during mitosis by Lyndon (1967) and with differ-

entation of cells by Lorz (1947) and Courtis (1964). Decreases in nu¬ clear size following irradiation have been reported by Schrek (1948). Nuclear size changes involve a gain or loss of water, of dry matter, or of some combination of the two.

This study was designed to de¬ termine the longitudinal distribu¬ tion pattern of nuclear size in bulbs of onion based on a model of cell aging, the youngest cells being at the base of the bulb and the oldest cells at the apex. Since nuclear size and nuclear dry mass were highly correlated (Courtis, 1964), it was considered desirable to characterize longitudinal distribution of nuclear size in leaf bases in order to estab¬ lish a basis for studies of cell senes¬ cence and death using quantitative interference microscopy.

In performing this study, we found that a number of bulbs ex¬ hibited fungal neckrots like those described by Walker (1968). In most cases, only the tip of the bulb apex was infected. Botrytis allii was easily isolated from many of these bulbs. The study reported herein includes a comparison of nuclear size in in¬ fected onions with nuclear size in non-inf ected ones. We assume the causal organism in the infected bulbs studied to be B. allii although no isolations were made from the tis¬ sue observed with the interference microscope.

Materials and Methods

Inner (adaxial) epidermis of ma¬ ture white onions, Allium cepa L., (approximately 10 cm in diameter) was used in a study of nuclear size

242

Kulfinski & Pappelis — Nuclear Size in Allium

243

distribution. The outer dry papery scales were removed, and the first exposed turgid scale was used in each of six onions. Ten samples were taken from a 5 mm wide lon¬ gitudinal strip extending from the stem at the base to the dead tissue at the apex of the bulb. Each sam¬ ple was one-tenth of the longitudi¬ nal strip (approximately 1 cm long), and these were numbered 1 through 10 from base to apex. The inner epidermis was removed from each location, mounted in tap water, and microscopically studied. Ten to twenty nuclei were selected in each location and photographed using a Leitz Orthomat camera on a Leitz interference microscope at a mag¬ nification of 500x. No attempt was made to reduce variation among nuclei such as selecting all the nu¬ clei from the same longitude or latitude within the sample area.

Only those nuclei lying against tan¬ gential faces of cells were studied since nuclei at the radial faces were seen in edge view rather than face view. Nuclear sizes were determined by tracing nuclear images from pro¬ photographic negatives and mea¬ suring the tracings with a planim- eter. Nuclear size observed in face view will be called nuclear area (NA) throughout this paper.

Fungal infections were confined to apical locations. Since the nu¬ cleus and cytoplasm of cells in con¬ tact with mycelium were usually disintegrated, host nuclei in the in¬ tact cells just ahead of the mycelial front were studied as well as those well in advance of this front. Aver¬ age NA of infected bulbs (minimum of five nuclei per replicate) were ex¬ pressed as a percentage of average NA from comparable locations of non-infected bulbs.

Table 1. — Mean nuclear area^/ in relation to location^/ in the outermost live bulb scale (inner epidermis) of onion.

Nuclear Area (10-6 cm2)

Location

Nuclear Area (10~6 cm2)

1

3.42

6

8.65

2

6.68

7

7.24

3

7.08

8

7.69

4

9.14

9

6.32

5

8.41

10

3.59

a. Six replicates, minimum of 10 nuclei per location per replicate.

b. Locations 1 through 10 are 10 equal distances from base to apex of the bulb scale.

Results

The location means (Table 1) in¬ dicate that nuclear areas in differ¬ ent locations of the bulb scale are not uniform but are distributed ac¬ cording to a pattern. The mean nu¬ clear area was least in location 1, generally increased to maximum in location 4, gradually declined to locations 7 and 8, and fell sharply from location 8 to 10. The nuclear areas of location 1 and 10 were nearly equal, and the greatest changes in nuclear areas were found

between locations 1 and 2 and be¬ tween 9 and 10.

An analysis of variance was per¬ formed which yielded F values of 4.54 and 16.28 for differences among replicate means and location means, respectively. Since the correspond¬ ing tabular values are 3.48 and 2.85 at the 1% level of probability (LeClerg, Leonard and Clark, 1962), then differences among both com¬ ponents are judged to be signifi¬ cant. Adjacent location means are compared using the Duncan mul-

244

Transactions Illinois Academy of Science

Table 2.— Duncan multiple range analysis of differences between adjusted lo¬ cation means. Differences greater than 16.60 are significant at the 5% level and those greater than 22.19 are significant at the 1% level of probability. The line separates the significant differences above from the non-significant below at the 1% level. Those marked * are significant at the 5% level.

Location

Location

4

6

5

8

7

3

2

9

10

1

66.67

63.30

60.40

51.57

48.97

49.29

39.40

35.07

1.84

10

64.83

61.46

58.56

49.73

47.13

42.45

37.66

33.23

9

31.60

28.23

25.33

16.50

13.90

9.22

4.33

2

27.27

23.90

21.00*

12.17

9.57

4.89

3

22.38

19.01*

16.11

7.28

4.68

7

17.70*

14.33

11.43

2.60

8

15.10

11.73

8.83

5

6.27

2.90

6

4

3.37

tiple range test (Table 2). The dif¬ ferences between locations 1 and 2, 3 and 4, and 9 and 10 are signifi¬ cant, whereas, the differences be¬ tween all other adjacent locations are non-significant at the 1% level of probability. Although other com¬ parisons can be made using Table 2, we would like to generalize as follows: no significant difference ex¬ ists in the average nuclear sizes of locations 1 and 10; little difference exists among the location means in the equatorial region; and nuclear sizes at the polar regions are signif¬ icantly smaller than those of the equatorial region.

In infected bulbs, the average NA in locations with mycelium and those adjacent to mycelium were found to be less than the NA aver¬ ages in the same locations in non- infected bulbs (Table 3). However, for cells more distant from the in¬ fection, average NA increased. In the case of mycelium present in the location, the greatest reduction in NA occurred in location 7 and least in 10. These also were the locations initially largest and least in average NA (for the three locations studied in which the mycelial front was observed). The reduction in NA in locations adjacent to those with

Table 3. — Mean nuclear area (NA) of inner epidermal nuclei adjacent to and distant from the mycelial front in neck-rotted white onion bulbs. Percentages are comparisons of the mean NA of infected with the mean NA of non-infected bulbs. Locations 1 through 10 are equal distances from base to apex of the outermost turgid bulb scale. Infections were in apical locations.

Location

Number

of

Replications

NA x 10"6 cm2 Infected

Percent

of

Non-infected

Infected

10

in

location 10.

4

2.9

81

9

4

6.1

97

Infected

9

in

locations 10 through 9.

2

3.2

51

8

1

5.5

71

Infected

7

in

locations 10 through 7.

2

3.4

47

6

2

5.6

64

Infected

in

10, 10 through 9, and 10 through 7, above.

6

8

9.7

111

1

8

3.6

106

Kulfinski & Pappelis — Nuclear Size in Allium

245

mycelium was similar to that de¬ scribed above; the amount of re¬ duction was related to the initial size. In all of the infected bulbs, the NA averages for locations 6 and 1 were greater than those in these locations in the non-infected bulbs.

Discussion

In the observed patterns of nu¬ clear sizes in non-infected bulbs, two biological interpretations are possible: nuclear size increases with cell enlargement during develop¬ ment; and nuclear size increases with distance from the basal meri- stematic zone and then declines with senescence and death (at the apex) after bulb maturation and harvest.

In the first interpretation, it is possible that nuclear area and other nuclear properties vary with phys¬ iological condition of cells based on their location, per se, rather than on distance from the meristematic zone at the base. If a cell-nuclear size relationship exists (Giese, 1962), then significant differences between adjacent location means would be anticipated near the poles and only slight differences near the equator.

In the second interpretation, the youngest cells in the onion bulbs are the basal ones (Hoffman, 1933). Nuclear areas increase by approxi¬ mately 2.7x from location 1 through 4. Some of the largest cells occur near the equator of the bulb, but it is not known if the increase in nuclear area is concommittant with cell enlargement. The bulb scales are leaf bases which either subtend or at one time subtended leaf blades. The drying and dying of blades at the top of a bulb begins in the field and ultimately results in dead bulb scale apices (Hoffman, 1933). It could be interpreted that the de¬ crease in mean nuclear area from location 4 through 10 may repre¬

sent a progressive deterioration dur¬ ing senescence in these locations after harvesting. Although the sharp difference in nuclear area between locations 9 and 10 may represent developmental differences, in all likelihood it represents progressive senescence and cell death since seg¬ ment 10 borders the apical dead zone.

The nuclear sizes in Table 1, the analysis of nuclear size means in Table 2, and our observation of largest cells in the equatorial region and smallest at the poles support both interpretations. Studies of the cell-nuclear size relationship during bulb development, pre-harvest, and post-harvest phases of growth are required to resolve this question.

Although the NA is known to in¬ crease or decrease in pathological conditions (Bessis, 1964; Cameron, 1951; Eckert and Cooper, 1937; Goodman, etal, 1967; Hasten, 1958; Kulfinski and Pappelis, 1969; Ogil- vie, 1962), the pattern of NA ob¬ served in onion epidermal cells ap¬ pears to be more related to normal growth and development processes. In this and other studies, we ob¬ served that NA of onion host cells decreased near the fungi and in¬ creased at some distance from the fungi (Kulfinski and Pappelis, 1969a, 1969b). In this study, the initial host NA varied due to lo¬ cation. We are assuming that the small nuclei in location 10 are less able to respond because they are in senescing cells. An additional test of this hypothesis would be to determine whether small non- senescing nuclei at the base of the bulb would be reduced in NA fol¬ lowing infection in their vicinity.

From the study of infected bulbs, we conclude that neckrot fungi lib¬ erate chemical factors (enzymes and/or toxins) at the site of the in¬ fection, which diffuse through the tissue away from the infection site

246

Transactions Illinois Academy of Science

and cause the nuclei of the host cells to decrease in size as a result of a combination of degradation of nuclear materials and changes in nuclear membrane character. The increase in N A at greater distances from the infection may be a re¬ sponse to metabolic changes in liv¬ ing host cells or to products from dead host cells nearer the infection. It appears that dying of host cells takes place very quickly in those cells in contact with the fungus and more slowly in cells more distant from the mycelium. It is also con¬ cluded, since host cell death in fun¬ gal parasitism and cellular autoly¬ sis (Kasten, 1958) have similar symptoms, that these degenerative changes and others as well may have common pathways, differing only in the specific form of initia¬ tion of the changes.

We conclude that the onion bulb epidermis represents a model in which to study nuclear control of cell development, senescence, death, and pathological changes. The fact that quantitative interferometric studies can be made on these living cells (Kulfinski and Pappelis, 1969a) without the obvious disruptions of morphology and composition which occur as a result of histological procedures contributes an impor¬ tant method to the study of the above-mentioned processes. Since NA correlates with nuclear dry mass (Courtis, 1964), and since there is a pattern of N A in the bulb scale, it may be that quantities of nuclear components vary with location and NA. Until the DNA content has been determined in these nuclei, an explanation of nuclear mass and NA changes can not preclude the possibility of endoploidy (Lorz, 1947). The determination of DNA, RNA, chromosomal histone, and total nuclear protein would be use¬ ful in the studies of development,

senescence, cell death, and cellular changes in response to pathogens.

Acknowledgements

This research was supported by Inter¬ disciplinary Research in Senescence, a Cooperative Project of Southern Illinois University. We wish to thank Dr. C. C. Lindegren for the use of the Leitz Inter¬ ference Microscope (P.H.S. Training Grant No. 5 TO GM 00593-08 from the National Institute of General Medical Science).

Literature Cited

Bessis, M. 1964. Studies on cell agony and death: an attempt at classification, pp. 287-316. IN. Cellular Injury, de Reuck, A.V.S., and J. Knight, Eds. Little, Brown, and Co., Boston, Mass. Cameron, G. R. 1951. Pathology of the Cell. Thomas, Springfield, Illinois. 840 p.

Courtis, W. S. 1964. An aging pattern of total nuclear dry mass in living epi¬ dermal cells of two monocotyledonous plants. M. S. Thesis. The University of Miami, Coral Gables, Fla. 48 p. Eckert, C. T., and Z. K. Cooper. 1937. Histologic study of nuclei in squamous cell carcinoma of the uterine cervix. Arch. Path. 24:476-480.

Giese, A. C. 1962. Cell Physiology. 2nd Ed. W. B. Saunders Co., Philadelphia, Pa. 592 p.

Goodman, R. N., Z. Kiraly, and M. Zaitlin. 1967. The Biochemistry and Physiology of Infectious Plant Disease. D. Van Nostrand Co., Inc., Princeton, New Jersey. 354 p.

Hoffman, C. A. 1933. Developmental morphology of Allium cepa. Bot. Gaz. 9:279-299.

Kasten, F. H. 1958. Nuclear size changes during autolysis in normal mouse liver, kidney, and adrenal glands. Proc. Soc. Exp. Biol, and Med. 98:275-277. Kulfinski, F. B., and A. J. Pappelis. 1969a. Changes in dry weights of onion nuclei in response to Botrytis allii. Phytopathology 59:115 (Abstr.) Kulfinski, F. B., and A. J. Pappelis. 1969b. Changes in size of onion nuclei in response to Botrytis allii. Phy¬ topathology 59:115. (Abstr.)

Leclerg, E. L., W. H. Leonard, and A. G. Clark. 1962. Field Plot Technique. 2nd Ed. Burgess Publishing Co., Min¬ neapolis, Minn. 373 p.

Lorz, A. P. 1947. Supernumary chromo¬ somal reproductions: polytene chromo¬ somes, endomitosis, multiple chromo¬ some complexes, polysomaty. Bot. Rev. 13:591-624.

Kulfinski & Pappelis — Nuclear Size in Allium

247

Lyndon, T. F. 1967. The growth of the nucleus in dividing and non-dividing cells of the pea root. Ann. Bot. N. S. 31:132-146.

Ogilvie, R. F. 1962. Histopathology. 6th Ed. Williams and Wilkin Co., Balti¬ more, Md. 514 p.

Schrek, R. 1948. Cytologic changes in

thymic glands exposed in vivo to x- rays. American Jour. Path. 24:1055- 1065.

Walker, J. C. 1968. Plant Pathology. 3rd Ed. McGraw-Hill Book Co., Inc., New York, N. Y. 819 p.

Manuscript received December 11+, 1970

VERSATILE APPARATUS FOR STUDYING REACTIONS INVOLVING GAS ADSORPTION OR EVOLUTION

JOSEPHUS THOMAS, JR., AND ROBERT R. FROST

Illinois State Geological Survey , Urbana , Illinois

Abstract. — Apparatus used for deter¬ mining surface area by a dynamic sorp¬ tion, or gas chromatography, method has great potential for other studies. Modifi¬ cations and techniques can transform the apparatus into a powerful research tool for studying various types of gas-solid interactions, including chemical or ther¬ mal decomposition and chemisorption. Evolved gas analysis (EGA) is the basis for most of the methodology.

Nelsen and Eggertsen (1958) de¬ scribed a dynamic gas adsorption method involving relatively simple apparatus for determining the sur¬ face areas of particulate solids. The method was further evaluated and refined by Daeschner and Stross (1962), and a commercial version of the apparatus, called the “Sorptom- eter,” was introduced shortly there¬ after by the Perkin-Elmer Corpora¬ tion (Norwalk, Conn.). Because of its similarity in operating princi¬ ples to those used in gas chroma¬ tography, this method for deter¬ mining surface area is commonly referred to as the chromatographic method. The surface area values determined with the classical BET (Brunauer, Emmett, and Teller, 1938) equation are in close agree¬ ment with those obtained by static methods in the more conventional pressure-volume apparatus.

With only slight modifications, the same apparatus is useful in con¬ siderably broader research studies. One of its major advantages, for example, is that the gaseous envi¬ ronment around a sample can be carefully controlled by adjusting the flow rates of gases or gas mix¬ tures passing over the sample. With the addition of a furnace and tem¬ perature controller, thermal decom¬ position studies can be conducted

at controlled temperatures and un¬ der controlled environments. A ther¬ mal conductivity detector is used for all gas measurements. The sen¬ sitivity and reliability of such a de¬ tector permit the use of small ( < 1 gram) samples, which is advanta¬ geous in decomposition studies.

The potential utility of this type of apparatus in physical research has not yet been fully realized or appreciated. This report, which is based on both published and un¬ published work from our labora¬ tory, calls attention to the versa¬ tility of the apparatus in studies involving gas-solid interrelation¬ ships.

Apparatus

A schematic diagram of the ap¬ paratus is shown in Figure 1. The basic gas flow scheme is the same as that described by Nelsen and Eggertsen (1958).

A thermal conductivity cell as¬ sembly (Gow-Mac 9193-TE-II) is contained in an oil bath at a con¬ trolled temperature (± 0.1° C). The reference and measuring detectors form part of well-known bridge cir¬ cuitry (Gow-Mac Bulletin TCThG 6-62-3M). A Sorensen QB12-2 d.c. power supply (Raytheon Company, South Norwalk, Conn.) provides a stable bridge current.

A Sargent Model MR (multi¬ range) recorder equipped with a Disc integrator (Disc Instruments, Inc., Santa Ana, Calif.) is used for recording bridge signals and inte¬ grating peak areas.

Most gases are delivered at less than 10 psig. Matheson Series 70 low pressure regulators are used for pressure control. Hoke precision

248

Thomas & Frost — Versatile Apparatus

249

Figure 1. Schematic diagram of apparatus used for various gas-solid studies.

needle valves (Enpro, Inc., Villa Park, Ill.) are used in conjunction with the low pressure regulators for controlling the gas flow rates. Corrosive gases, if desired, can be introduced into the inert carrier gas stream via calibrated loops of a gas sampling valve located up¬ stream from the sample. Gas mix¬ tures containing, for example, a known concentration (dilute) of a corrosive gas in an inert gas also can be purchased. If sufficiently dilute (and usually dry) such a mix¬ ture can be passed through the whole system with little danger of corrosion of the regulators or of the TC cell. The gas flow at any time is monitored by a rotameter and determined with a soap film flow¬ meter.

Our U-shaped sample holder is is made from Pt-Rh (5%) tubing and is approximately 12 inches long by 5/16 inch in diameter. To con¬

nect it to the rest of the apparatus, glass ball joints affixed to graded seals are sealed to the sample tube. Swagelok “quick connect” fittings also have been used successfully.

The furnace constructed in our laboratory is about 5-1/2 inches high and 7 inches in diameter over¬ all. The heated portion is an alumina cylinder 2-1/2 inches in diameter and closed at the bottom. The fur¬ nace, which has a handle attached, can be raised (or lowered) quickly into position around the sample holder. The temperature controller, also of our design, controls the fur¬ nace temperature to ± 0.5°C. Tem¬ perature is recorded directly by a Brown Electronik recorder, Model X-42A.

A Data-Trak, Model 5300, pro¬ grammer (Research, Inc., Minne¬ apolis, Minn.) has been found use¬ ful for temperature programming. With this unit a temperature pro-

250

Transactions Illinois Academy of Science

gram incorporating particular pauses at given temperatures or temperature intervals can be easily set up.

Uses of the Apparatus

Surface Area. The significance of this parameter should not be under¬ estimated. The surface of a solid differs from the rest of the substance in that the atoms or molecules at the surface are coordinatively un¬ saturated compared with the atoms or molecules below the surface. If surface area is increased by fine grinding, or if the surface area is large because of particle porosity, then the reactivity of the substance is usually greatly enhanced. For ex¬ ample, increased surface area gen¬ erally increases the rate of solu¬ bility, ion exchange, catalysis, cor¬ rosion, oxidation-reduction and thermal decomposition.

There is little need to discuss here either the operation of the ap¬ paratus or the calculations involved in surface area determinations. These details were given by Nelsen and Eggertsen (1958). Suffice it to say that the adsorbate (usually ni¬ trogen) is adsorbed near its boiling point at three relative pressures es¬ tablished by changing the flow rate of the adsorbate gas in the mixed gas stream. Helium acts as the car¬ rier gas. Adsorption, or desorption, of the adsorbate gas by the sample produces a peak on the recorder chart. The gas volume adsorbed (or desorbed) is determined from the peak area and from calibration curves relating peak area to known volumes of the adsorbate gas. Since detector response differs for the same volume of different gases, cali¬ bration is necessary for any gas that is introduced into or removed from a given gas stream.

Some of the more interesting sur¬ face area studies for which the ap¬ paratus is particularly well suited

involve the generation of surface within solid particles by thermal decomposition (Thomas, Hieftje, and Orlopp 1965). The rate of ther¬ mal decomposition, which also can be followed with the apparatus, is discussed in the next section. Sub¬ stances such as carbonates, nitrites, nitrates, hydroxides, carbonyls, and oxalates decompose thermally, evolving one or more gases and producing new solid phases that may be quite porous with conse¬ quent large internal surface area when decomposition is conducted under proper conditions. From nat¬ urally occurring carbonates, for ex¬ ample, calcium oxide and magne¬ sium oxide can be produced that have surface areas of about 60 m2/g and 500 m2/g, respectively. Because of the large available surface, such substances are often referred to as “active.”

A controlled environment is nec¬ essary for the study of “active” substances. Changes in the surface area of the “active” phase can be studied as a function of the sample environment (temperature or at¬ mosphere) either during the course of decomposition or after decompo¬ sition is complete. The differences that do occur involve crystal growth and sintering mechanisms. With the apparatus described, a sample can be decomposed, either wholly or to a specific partial extent, the surface area determined, and further ex¬ perimentation conducted to affect surface changes, all without moving the sample or exposing it to a del¬ eterious laboratory atmosphere.

Decomposition. “Active” solids with large surface areas are pro¬ duced readily, under some condi¬ tions, from solid-state thermal de¬ composition reactions of the type

A(solid) - B(solid) + C(gas or

gases). Most of these reactions con¬ tinue to completion under nonequi¬ librium conditions if the gas or

Thomas & Frost — Versatile Apparatus

251

gases are continually removed dur¬ ing a run, either by vacuum or by sweeping the sample with an inert gas such as helium. The rates of such reactions and the variables that influence the rates are of gen¬ eral interest in solid-state chemis¬ try. This type of study with the ap¬ paratus basically is evolved gas analysis (EGA).

In our apparatus a controlled constant flow of helium removes the gaseous decomposition product as

it evolves and carries it through the thermal conductivity cell for measurement. The decomposition is easily followed to completion on the recorder chart (Figure 2). The four different decomposition curves shown are for different sieve-size fractions of Iceland spar (calcite). All runs were made under the same experimental conditions. From the furnace-on position (850°C. initi¬ ally), about one minute elapses be¬ fore gas detection occurs.

Figure 2. Thermal decomposition of calcite cleavage pieces. Size fraction: A = 833-991 p, B = 295-417u, C = 74-89/*, D = <10/*.

Recorder data also can be pre¬ sented in the more common plot of oi (fraction decomposed) versus time (Figure 3). The fraction decom¬ posed (a) is determined from the integrator counts, as a fraction of the total integrator counts repre¬ senting complete decomposition, at time (t), which is determined from the recorder chart speed.

It is evident from these data that with decreasing crystallite size (not necessarily particle size) the rate of decomposition increases. This is well recognized for solids and is explained

by the fact that, in general, the more perfect a crystal is, the smaller its reactivity. The more reactive centers are found at places where defects or faults occur in the crystal order. As mentioned earlier, the surface of a crystal is one such place, as the outer-most, or surfi- cial, atoms are not bounded in three dimensions as are their neighbors immediately below the surface. Therefore, increasing the surface by subdividing the solid increases its reactivity, as is shown by the in¬ creased rate of thermal decompo-

252

Transactions Illinois Academy of Science

Figure 3. Fraction decomposed (£) versus time plots for decomposition of calcite (data from Figure 2).

sition. Defects within crystals may be caused by impurities, and they also can be introduced by irradia¬ tion.

The rate of thermal decomposi¬ tion also depends upon the tem¬ perature and upon the rate of re¬ moval of the gaseous products. Our apparatus readily lends itself to the study of the influence of these vari¬ ables on decomposition. For the past 10 years much of this type of research has been conducted with TGA (thermogravimetric analysis) apparatus, with which weight loss is directly recorded as a function of time at a given temperature. TGA is an excellent technique in this respect, but such apparatus is not well suited for low-temperature surface area measurements.

The modified dynamic sorption apparatus also can be used for stud¬ ies of chemical decomposition. Thomas and Hieftje (1966) de¬ scribed the use of the apparatus in the analytical determination of car¬ bon dioxide from carbonate-con¬

taining samples. The method is rapid and precise. It can be used ad¬ vantageously with small samples containing low ( < 1%) percentages of carbonate impurities. A sample tube with a side arm and rubber septum is used for injecting hydro¬ chloric acid onto the sample from a hypodermic syringe. The carbon¬ ate portion of the sample decom¬ poses, and helium carrier gas trans¬ ports the released carbon dioxide through the apparatus and into the thermal conductivity cell for mea¬ surement. Absorption tubes con¬ taining magnesium perchlorate and anhydrous copper sulfate are at¬ tached to the apparatus between the sample tube and detector to re¬ move water and undesirable acidic gases from the helium-carbon diox¬ ide stream.

Chemical Reactivity of Solids. A logical extension of the studies of decomposition and surface area is the study of chemisorption and chemical reactivity of solids. The sample tube functions as a reaction

Thomas & Frost — Versatile Apparatus

253

tube for the study of gas-solid re¬ actions at elevated temperatures on the “active” oxides formed. Carbon dioxide, for example, will recombine with “active” oxides of calcium and magnesium if equilibrium is shifted in favor of the carbonates. A study of the rate of recombination at vari¬ ous temperatures or at various par¬ tial pressures of carbon dioxide is possible with the apparatus.

Studies now in progress in our laboratory are concerned with the uptake of sulfur dioxide by “active” oxides of calcium and magnesium. A practical application of these studies is the removal of sulfur di¬ oxide, an air pollutant, from stack gases. One of the less expensive processes now being tested for re¬ moving sulfur dioxide involves the injection of limestones into a par¬ ticular portion of the hot zone of the stack.

In our studies a known volume of sulfur dioxide is introduced into the helium carrier gas by means of a gas sampling valve located up¬ stream from the sample. From the amount (by calibration) of sulfur dioxide that is measured at the de¬ tector, the relative efficiency of a particular oxide in removing sulfur dioxide from the gas stream and reacting with it to form a new solid phase can be determined. X-ray

diffraction is used to determine the nature of the new phase, or phases. The decomposition of the lime¬ stones, the surface area measure¬ ments, and the measurements on the efficiency of sulfur dioxide re¬ moval as a function of surface area and other variables, are all con¬ ducted on the sample in situ. To be able to control sample history as closely as is possible with this ap¬ paratus gives considerable confi¬ dence in the interpretation of re¬ sults.

Literature Cited

Brunauer, S., P. H. Emmet, and E. Teller. 1938. Adsorption of Gases in Multimolecular Layers. J. Am. Chem. Soc. 60:309-319.

Daeschner, H. W. and F. H. Stross. 1962. An Efficient Dynamic Method for Surface Area Determinations. Anal. Chem. 34:1150-1155.

Nelsen, F. M. and F. T. Eggertsen. 1958. Determinations of Surface Area. Anal. Chem. 30:1387-1390.

Thomas, J., G. M. Hieftje, and D. E. Orlopp. 1965. Application of Continu¬ ous-Flow Apparatus to Thermal De¬ composition and Sintering Studies. Anal. Chem. 37:762-763.

Thomas, J. and G. M. Hieftje. 1966. Rapid and Precise Determination of Carbon Dioxide from Carbonate-Con¬ taining Samples Using Modified Dy¬ namic Sorption Apparatus. Anal. Chem. 38:500-503.

Manuscript received February 10, 1971

CATALOG OF PALEOZOIC PALEOZOOLOGICAL TYPE AND FIGURED SPECIMENS AT THE ILLINOIS STATE MUSEUM

RICHARD L. LEARY, CURATOR

Illinois State Museum, Springfield, Illinois

Abstract. — The paleontology collec¬ tions of the Illinois State Museum con¬ tained numerous type and figured speci¬ mens. Over the years, many of these speci¬ mens have been transferred elsewhere re¬ sulting in much confusion as to the loca¬ tion of these important specimens. The following is a list of those now known to be in the Museum collections, exclusive of Pleistocene material.

The paleontology collections of the Illinois State Museum have, at one time or another, contained many type specimens and others figured in the literature. Dr. A. R. Crook (1911), Museum Director 1906 to 1930, wrote that the Museum pale¬ ontology collection contained 4700 described and 3500 figured speci¬ mens in 1910.

The most noteworthy single col¬ lection was that accumulated by A. H. Worthen (1813-1888), State Geologist of Illinois from 1858 until his death in 1888. Another signifi¬ cant collection is one built by George Langford, Sr. and his son, George Langford, Jr., which the Museum acquired in the late 1930’s. This collection is composed primarily of plant fossils although many inver¬ tebrate forms are also present.

Worthen, in collaboration with several paleontologists, published descriptions of many new species and figured other fossils in the Geo¬ logical Survey of Illinois (Volumes I-VIII) between 1866 and 1890. Other new species were first de¬ scribed in the Proceedings of the Academy of Natural Sciences, Phil¬ adelphia and later redescribed and figured in the Geological Survey of Illinois.

Later, portions of the collections were studied and the results pub¬

lished by various workers, among them Janssen (1939, 1940), Car¬ penter (1943), Raymond (1945), Petrunkevitch (1946), Kjellesvig- Waering (1948), Lowenstam (1948) and Langford (1958, 1963).

Many of the type and figured specimens formerly housed at the Illinois State Museum have over the years been transferred else¬ where. For example, many types were taken to the Illinois State Geo¬ logical Survey in Urbana, Illinois, during the 1930’s. Because of the resulting confusion regarding the locations of these important speci¬ mens, it was decided to locate and list as many as possible of those re¬ maining in the Museum’s collec¬ tions and to make this information available. Since their transfer into more accessible storage in the Mu¬ seum’s new building (1963), the collections have been searched for types and figured specimens.

Hanson and Scott (1967) pub¬ lished a catalog of Worthen types now located at the University of Illinois. Their catalog has been of great assistance to workers search¬ ing for these important materials.

In this catalog both ‘Type” and figured specimens are listed under the name used in the publication cited. In those instances in which a new species was figured in a publi¬ cation later than the original de¬ scription, this reference is also given.

In the Worthen collection, in par¬ ticular, specimens designated as ‘Type” may be holotype, syntype, paratype or, in a few cases, merely hypotypes. In those instances where it was possible to determine the

254

Leary — ISM Catalog

255

correct designation* it has been used. In other cases the original designa¬ tion, “type,” has been retained.

No attempt has been made to modernize the original author' s strat¬ igraphic or locality information. In nearly all cases this information is too generalized to permit improve¬ ment.

No effort was made to standard¬ ize references to formation, group or lithology. The same strata may be referred to as “group” in one de¬ scription and “limestone” in an¬ other. Locations may be given by county, the nearest city, an area such as “Mazon Creek area,” or in more general terms as, in the case of some figured specimens, “com¬ mon in southern Illinois.” The origi¬ nal terminology has been retained and, in the case of both stratigraphy and locality, must be understood as such.

The 66 specimens, representing 54 species, are distributed as fol¬ lows: Invertebrates: 48 species (27 types, 33 figured specimens); Ordo¬ vician-2, Silurian-13, Devonian-2, Mississippian-8, Pennsylvanian-35. Vertebrates: 6 species (5 types 1, figured specimen) ; Devonian-1, Mis- sissippian-5.

These specimens are available for examination at the Illinois State Museum; in some cases, loans may be arranged. Study of these and other specimens in the Museum's collections is not only welcomed but encouraged.

Abbreviations used in the

CATALOG ARE:

Co

County

Coal Meas

Coal Measures

Dev

Devonian

f

figure(s)

gr

group

la

Iowa

GSI

Geological Survey of

Illinois

Ill

Illinois

Ind

Indiana

ISMNHB

Illinois State Museum of Natural History Bulletin

ISM Sci Pap Illinois State Museum Scientific Papers

Is

limestone

Miss

Mississippian

Mo

Missouri

M&W

Meek and Worthen

N&W

Newberry and Worthen

no

number

Ord

Ordovician

P

page(s)

PA

Paleontographica

Americana

d1

Plate

ppa

Proceedings, Philadelphia Academy of Natural Science

pt

part

Sil

Silurian

ss

Sandstone

STJ&W

St. John and Worthen

U&E

Ulrich and Everett

v

volume

W&M

Worthen and Meek

The Catalog INVERTEBRATES PHYLUM ARTHROPODA Class Arachnoidea

Curculioides gracilis Petrunkevitch. Holo- type, ISM 14862. Petrunkevitch, 1946, ISM Sci Pap III, no 2, p 68-70, text- fig 34, pi 2, f 8-10. Penn. Mazon Creek,

Ill.

Discotarbus deplanatus Petrunkevitch. Figured specimen, ISM 14869. Pe¬ trunkevitch, 1913, Trans Conn Acad, v 18, p 121-122, textfig 75, 76, pi 12,

f 10. - , 1946, ISM Sci Pap III no 2,

p 23, pi 1, f 7. Penn. Mazon Creek, Ill.

Euproops danae M&W. Figured specimen, ISM 14808. Raymond, 1945, ISM Sci Pap III no 3, p 4-6, pi 2, f 1. Francis Creek sh, Penn. Grundy Co., Ill.

E. danae M&W. Figured specimen, ISM 14812. Raymond, 1945, ISM Sci Pap III no 3, p 4-6, pi 2, f 1. Francis Creek sh, Penn. Grundy Co., Ill.

E. thompsoni Raymond. Figured speci¬ men, ISM 14807. Raymond, 1945, ISM Sci Pap III no 3, p 4-6. Francis Creek sh, Penn. Grundy Co., Ill.

Lepidoderma mazonense M&W. Figured specimen, ISM 14813. Kjellesvig-Waer- ing, 1948, ISM Sci Pap III no 4, p 17-24, pi 2, f 3, 4. Francis Creek sh, Penn. Will Co., Ill.

L. mazonense M&W. Figured specimen, ISM 14814. Kjellesvig-Waering, 1948, ISM Sci Pap III no 4, p 17-24, pi 3, f 1. Francis Creek sh, Penn. Will Co., Ill.

256

Transactions Illinois Academy of Science

L. mazonense M&W. Figured specimen, ISM 14815. Kjellesvig-Waering, 1948, ISM Sci Pap III no 4, p 17-24, pi 3, f 2. Francis Creek sh, Penn. Will Co., Ill.

L. mazonense M&W. Figured specimen, ISM 14816. Kjellesvig-Waering, 1948, ISM Sci Pap III no 4, p 17-24, pi 2, f 1, 2. Francis Creek sh, Penn. Will Co., Ill.

Ootarbus ovatus Petrunkevitch. Holotype, ISM 14866. Petrunkevitch, 1946, ISM Sci Pap III no 2, p 41, 42, textfig 23, pi 1, f 5, 6. Francis Creek sh, Penn. Mazon Creek, Ill.

0. pulcher Petrunkevitch. Type, ISM 14861. Petrunkevitch, 1946, ISM Sci Pap III no 2, p 36-40, textfig 15-20, pi 3, f 12, 13. Francis Creek sh, Penn. Mazon Creek, Ill.

O. pulcher Petrunkevitch. Paratype, ISM 14870. Petrunkevitch, 1946, ISM Sci Pap III no 2, p 40, 41, textfig 21, 22, pi 3, f 17. Francis Creek sh, Penn. Mazon Creek, Ill.

O. pulcher Petrunkevitch. Figured speci¬ men, ISM 14868. Petrunkevitch. 1946, ISM Sci Pap III no 2, p 39, 40, pi 4, f 22. Francis Creek sh, Penn. Mazon Creek, Ill.

Orthotarbus robustus Petrunkevitch. Type, ISM 14863. Petrunkevitch, 1946, ISM Sci Pap III no 2, p 29-32, textfig 9-11, pi 3, f 14, 15. Francis Creek sh, Penn. Mazon Creek, Ill.

0. robustus Petrunkevitch. Paratype, ISM 14865. Petrunkevitch, 1946, ISM Sci Pap III no 2, p 32, 33, pi 3, f 16. Fran¬ cis Creek sh, Penn. Mazon Creek, Ill.

0. robustus Petrunkevitch. Paratype, ISM 14867. Petrunkevitch, 1946, ISM Sci Pap III no 2, p 32, 33, textfig 12, 13, pi 4, f 20, 21. Francis Creek sh, Penn. Mazon Creek, Ill.

0. robustus Petrunkevitch. Figured speci¬ men, ISM 14874. Petrunkevitch, 1946, ISM Sci Pap III no 2, p 34-, pi 4, f 18, 19. Francis Creek sh, Penn. Mazon Creek, Ill.

O. robustus Petrunkevitch, Figured speci¬ men, ISM 14878. Petrunkevitch, 1946, ISM Sci Pap III no 2, p 34-36, textfig 14, pi 4, f 23. Francis Creek sh, Penn. Mazon Creek, Ill.

Paratarbus carbonarius Petrunkevitch. Holotype, ISM 14864. Petrunkevitch, 1946, ISM Sci Pap III no 2, p 25-27, textfig 5, 6, pi 2, f 11. Francis Creek sh, Penn. Mazon Creek, Ill.

Pleophrynus ensifer Petrunkevitch, Holo¬ type, ISM 14873. Petrunkevitch, 1946, ISM Sci Pap III no 2, p 52-58, textfig

25-27, pi 1, f 1-4. Francis Creek sh, Penn. Mazon Creek, Ill.

Class Insecta

Architarbus rotundatus Scudder. Figured specimen, ISM 14871. Scudder, 1868, GSI, v 3, p 568, textfig 4. Petrunke¬ vitch, 1913, Trans Conn Acad, v 18, p 125-128, textfig 79, 80, pi 12, f 72-75,

pi 13, f 76-78. - , 1946, ISM Sci Pap

III, no 2, p 20, 21, textfig 1. Coal Meas, Penn. Mazon Creek, Grundy Co., Ill.

Heterologus langfordorum Carpenter. Ho¬ lotype, ISM 14879. Carpenter, 1943, ISM Sci Pap III no 1, p 15, 16, textfig

4, pi 3. Francis Creek sh, Penn. Will Co., Ill.

Lithoneura mirifica Carpenter. Holotype, ISM 14880. Carpenter, 1943, ISM Sci Pap III no 1, p 13, 14, textfig 3, pi 2. Francis Creek sh, Penn. Will Co., Ill.

Syntonoptera schucherti Handlirsch. Fig¬ ured specimen, ISM 14881. Carpenter, 1943, ISM Sci Pap III no 1, p 12, text¬ fig 2. Francis Creek sh, Penn. Will Co., Ill.

Teneopteron mirabile Carpenter. Holo¬ type, ISM 14887. Carpenter, 1943, ISM Sci Pap III no 1, p 17-20, textfig

5, pi 4. Francis Creek sh, Penn. Will Co., Ill.

Thesoneura americana Carpenter. Holo¬ type, ISM 14876. Carpenter, 1943, ISM Sci Pap III no 1, p 10, 11, pi 1, f 1, Francis Creek sh, Penn. Will Co., Ill.

Class Trilobita

Acidaspis hamata Conrad. Figured speci¬ men, ISM 2202, M&W, 1868, GSI, v 3, p 390, pi 7, f 15. Lower Helderberg, Dev. Perry Co., Mo.

Illaenus crassicauda Wahlenberg. Figured specimen, ISM 12031. M&W, 1868, GSI, v 3, p 322, pi 3, f 1. Galena gr., Ord. Galena, Ill.

PHYLUM BRACHIOPODA

Athyris subtilita Hall. Figured specimen, ISM 2939. M&W, 1873, GSI, v 5, p 570, pi 25, f 14. Coal Meas., Penn. La Salle Co., Ill.

Orthis carbonaria Swallow. Figured speci¬ men, ISM 2939. Worthen, 1873, GSI v 5, p 571, pi 25, f 4. Upper Coal Meas., Penn. LaSalle, Ill.

Productus longispinus Sowerby. Figured specimen, ISM 2927. M&W, 1873, GSI v 5, p 569, pi 25, f 10. Coal Meas., Penn., Ill.

Spirifer cameratus Morton. Figured speci¬ men, ISM 8491. M&W, 1873, GSI, v 5, p 573, pi 25, f 6. Coal Meas., Penn.

Leary — ISM Catalog

257

PHYLUM BRYOZOA (POLYZOA)

Archimedes grandis Ulrich. Type, ISM 2861. Ulrich, 1890, GSI, v 8, p 569, pi 63, f 10. Keokuk gr., Miss. Jersey Co.,

III.

PHYLUM COELENTERATA

Zaphrentis centralis Edwards & Haime. Figured specimen, ISM 2569. Worthen, 1890, GSI, v 8, p 72, pi 9, f 2. Burling¬ ton Is, Miss. Henderson Co., Ill.

PHYLUM ECHINODERMA

Alisocrinus? heterodactylus Brower. Para- type, ISM 1565. Brower, 1970, PA, in press. Girardeau Is, Ord. Orchard Creek, Alexander Co., Ill.

Barycrinus hoveyi Hall. Figured specimen, ISM 1617. M&W, 1873, GSI, v 5, p 486, pi 13, f 1. Keokuk gr. Miss. Craw- fordsville, Ind.

B. hoveyi var herculeus M&W. Type, ISM 1805. M&W, 1868, PPA 1868, p 341.

- , 1873, GSI, v 5, p 485, pi 13, f 2a,

2b, Keokuk, gr, Miss. Crawfordsville, Ind.

Eucalyptocrinus lindahli Wachsmuth & Springer. Type, ISM 10467. W&S,

1892, Amer Geol v 10, p 139. - ,

1897, Mem Mus Com Zoo Harvard, v 20, p 347, pi 82. E. wortheni; Miller & Gurley, 1893, ISMNHB no 3, p 53, pi 4, f 2. Niagaran, Sil. Wayne Co., Tenn.

Eucalyptocrinus sp. Figured specimen, ISM 15976. Lowenstam, 1948, ISM Sci Pap IV, pi 6, f 3, Blue Island Zone, Sil., Ill.

Eupachycrinus boydi M&W. Holotype, ISM 2443. M&W, 1870, PPA, p 30.

- , 1873, GSI v 5, p 554, E. Aspera-

tus Worthen. Worthen 1882, ISMNHB

1, p 34, - , 1883, GSI v 7, p 311, 312,

pi 29, f 4. Chester gp. Miss. Monroe Co., Ill.

Myelodactylus sp. Figured specimen, ISM 15977. Lowenstam, 1948, ISM Sci Pap

IV, pi 6, f 7, Blue Island Zone, Sil. Blue Island, Ill.

Pisocrinus benedicti S. A. Miller. Figured specimen, ISM 15972. Lowenstam, 1948, ISM Sci Pap IV. pi 5, f 4, 5. Lis¬ ton Creek fm, Sil. Wabash, Ind.

P. campana S. A. Miller. Figured speci¬ men, ISM 15978. Lowenstam, 1948, ISM Sci Pap IV, pi 6, f 8. Racine- Guelph, Sil. Thornton, Ill.

P. quinquelobus Bather. Figured speci¬ men. ISM 15973. Lowenstam, 1948, ISM Sci Pap IV, pi 5, f 7. Racine- Guelph, Sil. Thornton, Ill.

Scaphiocrinus carbonarius M&W. Figured specimen, ISM 1908. M&W, 1873, GSI, v 5, p 562, pi 24, f 2. Coal Meas., Penn. Springfield, Ill.

Zeacrinus ( Hydreionocrinus ?) acantho- phorus M&W. Type, ISM 1906. M&W,

1870, PPA, p 28. - , 1873, GSI, v 5,

p 563-565, pi 24, f 11. roof of coal no 1, Penn. Seaville, Fulton Co., Ill.

PHYLUM MOLLUSCA Class Gastropoda

Loxonema cf. leda Hall. Figured specimen, ISM 15979. Lowenstam, 1948, ISM Sci Pap IV, pi 7, f 6-8. Dalmanites Zone, Sil. Blue Island, Ill.

Class Pelecypoda

Allorisma costata M&W. Type, ISM 2975. M&W 1869, PPA, p 171. — , 1873, GSI, v 5, p 585, 586, pi 26, f 15. Lower Coal Meas., Penn. Warren Co., Ill.

A. elongata Worthen. Type, ISM 2542. Worthen, 1884, ISMNHB no 2, p 12.

- , 1890, GSI, v 8, p 133, pi 19, f 10.

Keokuk Is, Miss. Warsaw, Ill.

A. illinoiensis Worthen. Type, ISM 2540. Worthen, 1884, ISMNHB no 2, p 11.

- , 1890, GSI, v 8, p 132, pi 18, f 1,

la. Keokuk Is, Miss. Warsaw, Ill.

Bakevellia illinoiensis Worthen. Type, ISM 2525. Worthen, 1884, ISMNHB

no 2, p 14. - , 1890, GSI, v 8, p 126,

pi 18, f 4, 4a. Upper Coal Meas. Penn. LaSalle Co., Ill.

Grammysia rhomboidalis M&W. Type, ISM 4603. M&W, 1865, PPA, 1865, p

248. - , 1868, GSI, v 3, p 439, pi 11,

f 3a, 3b. Hamilton gr, Dev. Jackson Co., Ill.

Macrodon sangamonensis Worthen. Type, ISM 2585. Worthen, 1890, GSI, v 8, p 123, pi 21, f 3. Coal Meas., Penn. Ralls Ford, Sangamon Co., Ill.

Schizodus varsoviensis Worthen. Type, ISM 2500. Worthen, 1884, ISMNHB

no 2, p 10. - , 1890, GSI, v 8, p 107,

pi 19, f 7. Keokuk gr., Miss. War¬ saw, Ill.

PHYLUM PORIFERA

Anthaspidella scutula U&E. Type, ISM 2639. U&E, 1890, GSI, v 8, p 261, pi 3, f 1, la. Trenton Is, Sil. Dixon, Ill.

Astraeospongia meniscus Roemer. Figured specimen, ISM 15967. Lowenstam, 1948, ISM Sci Pap IV, pi 1, f 11, 12. Liston Creek fm, Sil. Wabash, Ind.

A. meniscus Roemer. Figured specimen, ISM 15968. Lowenstam, 1948, ISM Sci Pap IV, pi 2, f 5, 6. Blue Island Zone, Sil. Blue Island, Ill.

258

Transactions Illinois Academy of Science

A. meniscus Roemer. Figured specimen, ISM 15969. Lowenstam, 1948, ISM Sci Pap IV, pi 3, f 7-9. Blue Island Zone, Sil. Blue Island, Ill.

A. mensicus Roemer. Figured specimen, ISM 15971. Lowenstam, 1948, ISM Sci Pap IV, pi 5, f 1, 2. Blue Island Zone, Sil. Blue Island, Ill.

Astylospongidae sp. indet. Figured speci¬ men, ISM 15970. Lowenstam, 1948, ISM Sci Pap IV, pi 4, f 6. Waukesha fm, Sil. Elmhurst, Ill.

VERTEBRATES

PHYLUM CHORDATA Subphylum Vertebra ta

Class Pices

Cochliodus Leidyi StJ&W. Type?, ISM 12824. StJ&W, 1883, GSI, v 7, p 127- 130. Chester Is, Evansville, Ill.

Dinichthys ( Eastmanosteus ) pustulosus Eastman. Type, ISM 416238. Eastman, 1902, Am Nat, v 36, n 428, p 653-657, f 1. Hamilton Is, Dev. Andulusia, Rock Island, Co., Ill.

Physonemus falcatus StJ&W. Type, ISM 8720. StJ&W, 1883, GSI, v 7, p 252, pi 24, f 6. St. Louis Is, Miss., St. Louis, Mo.

Pnigeacanthus trigonalis StJ&W, Type?, ISM 7179, StJ&W, GSI, v 7, p 259, 260. St. Louis Is, Miss., Alton, Ill.

Psammodus crassidens StJ&W. Type, ISM 7083. StJ&W, 1883, GSI, v 7, p 218, pi 18, f 6. St. Louis Is, Miss. St. Louis, Mo.

P. Plenus StJ&W. Type, ISM 7154. StJ&W, 1883, GSI, v 7, p 213, pi 16, f 4. St. Louis Is, Miss. St. Louis, Mo.

Literature Cited

Brower, J. C. (in press) Paleontograph- ica Americana monograph.

Carpenter, F. M. 1943. Carboniferous Insects from the vicinity of Mazon Creek, Ill. Ill. State Mus. Sci. Pap. Ill, no. 1, p. 1-24, 5 textfigures, 4 plates.

Crook, A. R. 1911. Report on the prog¬ ress and condition of the Ill. State Mus. of Nat’l. Hist, for the years 1909 and 1910. Springfield, Ill.

Hansman, R. H. and Scott, H. W. 1967. Catalog of Worthen type and figured specimens at the University of Ill. Jour. Paleo. 41 (4), p. 1013-1028.

Janssen, R. E. 1939. Leaves and Stems from Fossil Forests. Ill. State Mus. Pop. Sci. series, vol. 1, 190 pp. 165 figs.

Kjellesvig-Waering, E. N. 1948. The Mazon Creek Euripterid. Ill. State Mus. Sci. Pap. Ill, no. 4, p. 1-64, 8 plates.

Langford, George 1958. The Wilming¬ ton Coal Flora. ESCONI Assoc., Downers Grove, Ill. 360 pp., 674 figs. _ 1963. The Wilmington Coal

Fauna and additions to the Wilming¬ ton Coal Flora. ESCONI Assoc., Downers Grove, Ill. 280 pp., 471 figs.

Lowenstam, H. A. 1948. Biostratigraphic Studies of the Niagaran Inter-reef For¬ mations in northeastern Illinois. Ill. State Mus. Sci. Pap. IV, 146 pp., 7 plates.

Meek, F. B. and Worthen, A. H. 1865, 1868. Proceedings , Acad, of NatT. Sciences, Philadelphia.

_ 1868. Paleontology of Illinois.

Geol. Surv. of Ill. Ill, p. 289-565.

_ 1869, 1870. Proceedings. Acad.

NatT Sci., Philadelphia.

_ _ 1873. Paleontology of Illinois.

Geol. Surv. of Ill. V, p. 323-619.

Miller, S. A. and Gurley, Wm. F. E. 1893. Description of some new species of invertebrates from the Paleozoic rocks of Illinois and adjacent states. Ill. State Mus. of NatT Hist. Bull. no. 3, 81 pp., 8 pis.

Petrunkevitch, Alexander 1913. A monograph of the Terrestrial Paleozoic Arachnida of North America. Trans. Conn. Acad. Vol. 18.

_ 1946. Paleozoic Arachnida of

Illinois. Ill. State Mus. Sci. Pap. Ill, no. 2, p. 1-76, textfigs., 4 plates.

Raymond, P. E. 1945. Xiphosura in the Langford collection. Ill. State Mus. Sci. Pap. Ill, no. 3, p. 1-10, 2 plates.

Scudder, S. H. 1868. Descriptions of fos¬ sil insects. Geol. Surv. of Ill. Ill p. 566-572.

Ulrich, E. O. and Everett, Oliver 1890. Descriptions of Lower Silurian sponges. Geol. Surv. of Ill. VIII, p. 253-282.

Ulrich, E. O. 1890. Paleozoic Bryozoa. Geol. Surv. of Ill. VIII p. 283-678.

Wachsmuth, C. and Springer, F. 1892. Amer. Geol. V. 10, p. 139.

_ _ _ _ _ _ . 1897. Mem. Mus. Comp.

Zoo., Harvard V. 20, p. 247.

Worthen, A. H. 1882. Descriptions of fifty-four new species of Crinoids from the Lower Carboniferous Limestones and Coal Measures of Ill. and Iowa. Ill. State Mus. of NatT. Hist. Bull. I, p. 1-38.

Leary — ISM Catalog

259

_ 1883. Descriptions of fossil

invertebrates. Geol. Surv. of Ill. VII, p. 267-338.

_ 1884. Descriptions of two

new species of Crustacea, fifty-one spe¬ cies of Mollusca, and three species of Crinoids, from the Carboniferous For¬

mation of Ill. and adjacent states. Ill. State Mus. of Nat’l. Hist., Bull. 2, 27 pp.

_ 1890. Description of fossil

vertebrates. Geol. Surv. of Ill. VIII, p. 69-154.

Manuscript received February 8, 1971.

THE CYPERACEAE OF ILLINOIS. XII. CAREX, PART 1

DAN K. EVANS AND ROBERT H. MOHLENBROCK

Southern Illinois University, Carbondale

Abstract. — This is the initial article in a series on the systematics of the genus Carex in Illinois. Treated in this paper are the sections Divisae, Chordorrhizeae, and Arenariae. A new variety is named. Keys and descriptions are provided.

This is the first in a series of pub¬ lications dealing with the genus Carex in Illinois. The other eleven genera of Cyperaceae in Illinois have been treated in prior issues of The American Midland Naturalist and The Transactions of the Illinois State Academy of Science. The il¬ lustrations are by Fredda Burton. One or more sections of Carex will be the subject of each of the papers. Keys to various groups will be pro¬ vided when necessary. A key to all taxa of Carex in Illinois will follow the systematic account of the taxa.

Species of Carex are perennials with grass-like leaves. Their stems are frequently triangular and bear 3-ranked leaves. The leaves are composed of a blade, a sheath, and a ligule.

The flowers are borne in spikes or heads in the axils of bracts (scales). The flowers are unisexual and borne either in different parts of the spikes or in separate spikes on the same culm. Rarely are the plants dioe¬ cious.

. Neither the staminate nor the pistillate flower has a perianth. The staminate flower merely consists of three stamens in the axil of a scale. The pistillate flower consists of one pistil which is enclosed in a sac (perigynium) in the axil of a scale. The style is either 2- or 3-cleft.

The fruit is an achene enclosed in the perigynium. It may be lenticu¬ lar, plano-convex, or trigonous.

Carex in Illinois is generally con¬ sidered to be divided into two sub¬ genera. Subgenus Vignea has mostly

uniform and sessile spikes, two styles, and a lenticular or plano¬ convex achene. Subgenus Carex has at least some all-pistillate spikes, usually three styles, and a trigonous achene.

The Illinois species of Carex fall into eleven sections of subgenus Vignea and twenty-seven sections of subgenus Carex.

This paper is concerned with sec¬ tions Divisae, Chordorrhizeae, and Arenariae of subgenus Vignea.

In addition to having the usual characters of the subgenus, the taxa of these three sections all possess slender, well-developed rhizomes, have some or all spikes androg¬ ynous, and are monoecious.

Systematic Treatment §Divisae

Rhizomatous perennials; culms mostly solitary ; spikes androgynous, distinct or more often in interrupted or subcontinuous heads, the lowest bracts awned, awn-tipped, or bear¬ ing a short blade; styles 2; perigynia coriaceous, usually with a narrow margin, with an entire or serrulate short beak; achenes lenticular or plano-convex.

Key to the Taxa of § Divisae in Illinois

1. Rootstock wiry, with profuse fibrous roots; culms to 15 cm tall; leaves stiffly erect, to 12 cm long, with sharply tri¬ angular tips; inflorescence head-like, 0.8-1. 3 cm long; perigynia 3-7 per spikelet, with wrinkled striations ven- trally . . . . 1. C. stenophyllci var. enervis

1 . Rootstock stout, with few fibrous roots ; culms to 65 cm tall; leaves ascending to spreading, to 35 cm long, flattened throughout; inflorescence spike-like, 2-4 cm long; perigynia 8-12 per spike-

260

Evans & Mohlenbrock — Carex of Illinois

261

Figure 1. Carex stenophylla var. enervis. a. Habit, x 1/2. b. Inflorescence, x 1-1/2. c. Map. d. Inflorescence, x 1-1/2. e. Staminate scale, x 7-1/2. f. Pistillate scale, x 7-1/2. g. Perigynia, dorsal view, x 7-1/2. h. Perigynia, ventral view, x 7-1/2. i. Achene, x 7-1/2.

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let, without ventral nerves and wrin¬ kles . 2. C. praegracilis

1. Car ex stenophylla Wahlenb. var. enervis (C.A.Mey.) Kiikenth. in Engl. Das

Pflanzenreich 4(20) :122. 1909. Fig. 1.

Carex enervis C.A.Mey. in Ledeb. FI.

Altaica 4:29. 1833.

Carex eleocharis Bailey, Mem. Torrey

Club 1:6. 1889.

Plants cespitose, from fibrous roots with scaly, wire-like rootstocks; culms slender, smooth, canaliculate, (3-) 6-15 cm tall, slightly exceeding the leaves with the base fibrillose; leaves 2-4, (2-) 6-12 cm long, 0.75-1.50 mm wide, arising near the base, canaliculate, ascending, becom¬ ing triangular at the tip with the margins subscabrous; sheath apex truncate, the old sheaths persistent; inflorescence an¬ drogynous, 8-13 mm long, 3-5 mm wide; spikelets 4-7, scarcely distinguishable; pis¬ tillate scales ovate, 2. 5-3. 5 mm long, 1.5- 2.0 mm wide, acuminate, reddish-brown, margins hyaline, completely concealing the perigynia; staminate scales narrow- lanceolate, 3-4 mm long, 1.0-1. 5 mm wide, acuminate, reddish-brown in upper half, hyaline in the lower half; bracts encircling the culm with the lowest awn-tipped; perigynia 3-7 per spikelet, 2. 5-3.0 mm long, 1.25-1.50 mm wide, plano-convex, ovate-acuminate, the margins elevated ventrally with the upper half serrate, obscurely striate ventrally and dorsally, greenish-brown, substipitate, finely retic¬ ulate throughout with the bidentate beak 1 mm long, serrate, hyaline-tipped and obliquely cleft dorsally; achene 1.50-1.75 mm long, 1.25-1.50 mm wide, lenticular, yellow-green, finely puncticulate, jointed to a short style; stigmas 2.

Habitat: Gravel-bluff prairie.

Range: Manitoba northwest to Yu¬ kon, south to eastern Oregon, Utah, New Mexico, Iowa, and northern Illinois.

Both Eurasian and American material were originally called Carex stenophylla Wahlenberg. However, Eurasian material tends to have perigynia 3. 0-3. 5 mm long and nerved, while American material has perigynia 2. 5-3.0 mm long and essentially nerveless. American material was first segregated as Carex enervis C.A. Mey., then as Carex stenophylla var. enervis (C.A. Mey.) Ktikenth., and finally as Carex eleocharis Bailey.

Carex stenophylla var. enervis is best distinguished by its leaf shape. The leaf tip becomes triangular, stiff, and sharp- pointed, accounting for its common name, the Needleleaf Sedge. Its short, erect stature, fibrous roots with wiry rootstock, and nerveless perigynia provide good iden¬ tifying characters as well.

Although Fernald (1950) remarks that the spikelets appear in a definitely in¬ terrupted head, the Illinois material ex¬ amined had spikelets that were scarcely distinguishable from each other..

Carex stenophylla var. enervis is pri¬ marily a western and northern sedge and can be found in abundance in the mixed prairie states. Hanson and Churchill (1961) remark that C. eleocharis is con¬ sidered a dominant in the blue grama- grass-needlegrass-sedge communities on the upland plateaus of western North Dakota. Weaver and Albertson (1956) re¬ port that this taxon ranks high among the species of the mixed prairie where it oc¬ curs in dense patches and provides early grazing.

This taxon is rare in Illinois, the east¬ ernmost limit of its range, and may be limited to Winnebago County where Fell (1958) reports the only known location.

Specimens Examined: WINNEBAGO: Camp Grant, S. of Rockford, gravel-bluff prairie, April 29, 1957, Fell 57-9 (ISM, ILL); Camp Grant, S. of Rockford, gravel-bluff prairie, May 6, 1957, Fell 57-68 (ISM, ILL); Camp Grant, S. of Rockford, May 18, 1957, Fell 57-157 (ISM, ILL); Camp Grant, S. of Rockford, gravel-bluff prairie, May 26, 1957, Fell 57-21+8 (ISM, ILL).

2. Carex praegracilis W. Boott in Coult.

Bot. Gaz. 9:87. 1864. Fig. 2.

Plants from stout, horizontal, brown to black, scaly and fibrillose rootstocks; culms slender, sharply angled and sca¬ brous above, less angled and smooth be¬ low, (15-) 30-65 cm tall, usually equalling or slightly exceeding the upper leaves, the base scaly; leaves 2-4, 5-35 cm long, flat¬ tened, canaliculate, 2-3 mm broad, as¬ cending to spreading with the margins scabrous; sheath apex truncate, the old sheaths persistent; inflorescence androg¬ ynous, narrowly cylindric to slightly spreading, 2-4 cm long, 3-5 mm broad; spikelets 5-12, the upper ones crowded, interrupted in the lower half; pistillate scales ovate, 3. 5-4.0 mm long, 1. 5-2.0 mm broad, acuminate, pale to brown with a darker keel, margins hyaline, concealing the perigynia; staminate scales narrowly lanceolate, inconspicuous, 3-4 mm long, 1-2 mm broad, acuminate, pale, margins hyaline; bracts lanceolate, the lowest en¬ larged and scabrous-awned, becoming smaller and less awned toward the apex, often encircling the culm with a dark ring at the base; perigynia 8-12 per spikelet, 2.8-3.75 mm long, 1.25-1.50 mm broad, plano-convex, stipitate, ovate-lanceolate, sparsely nerved dorsally, nerveless ven¬ trally, thin-winged and serrulate above, brownish-black with the 1 mm long serru-

Evans & Mohlenbrock — Carex of Illinois

263

Figure 2. Carex praegracilis. a. Habit, x 1. b. Inflorescence, x 1. c. Map. d. Stami- nate scale, x 7-1/2. e. Pistillate scale, x 7-1/2. f. Perigynia, dorsal view, x 7-1/2. g. Perigynia, ventral view, x 7-1/2. h. Achene, x 7-1/2.

264

Transactions Illinois Academy of Science

late beak cleft dorsally; achene 1. 5-2.0 mm long, 0.75-1.25 mm broad, lenticular, dark brown, polished, finely puncticulate, jointed to a short style; stigmas 2.

Habitat: Low prairies, roadsides, and dry sterile soil.

Range: Manitoba northwest to Yu¬ kon, south to Mexico, Missouri, Iowa, northern Illinois, and northern Michigan.

Carex praegracilis W. Boott is a most variable species and without striking char¬ acteristics. The inflorescence may be lin- ear-cylindric with appressed spikelets or with a short-spreading head. The color of the inflorescence varies from pale to dark brown. The species, though usually tall, slender, and sometimes lax, may also be short and stiffly erect. Spongberg 61/38 and 61/38A collected from Bell Bowl Prairie show evidence of burning and therefore the short, erect stature of that material may be anomalous.

Carex praegracilis, especially immature material, may be confused with C. sar- twellii Dew. or C. foenea Willd. However, the combination of brownish-black perig- ynia without ventral nerves and the often tall, triangular and scabrous culms pro¬ vides identifying characters.

Fernald (1950) equates Carex marcida Boott with G. praegracilis; however, Boott’s illustration of C. marcida shows a thicker inflorescence and a markedly winged perigynia. Such characters are not present in Illinois material.

Carex praegracilis, found commonly in the western states and Canada, is re¬ ported from only two areas east of the Mississippi River, northern Illinois and northern Michigan. In Illinois this very rare sedge is limited to Winnebago, Kane, and DeKalb counties.

Specimens Examined: DEKALB: Near Kingston, June 8, 1953, Fell 53371 (ISM) ; W. edge of Kingston, June 2, 1956, Fell 56-91 (ILL); W. edge of Kingston, May 13, 1955, Fell 55-1/5 (ILL.) KANE: El¬ gin, April 27, 1919, Benke 3591 (F) [pre¬ viously determined as C. siccata Dew.]. WINNEBAGO: Camp Grant, June 14, 1955, Fell & Fuller 17200 (ILL, ISM, SIU); Camp Grant, August 2, 1953, Fell 53856 (ISM) [originally determined C. sartwellii Dew.]; S. of Rockford, May 21, 1952, Fell 52105 (ISM) ; Bell Bowl Prairie, May 10, 1961, Spongberg 61/38, 61/38A, 61/ /l, 61/ /I A (ISM); Perryville Road, June 6, 1955, Fell 55-//0 (ILL); Camp Grant, May 29, 1955, Fell 55-318 (ILL); Camp Grant, June 5, 1956, Fell 56-113 (ILL); Perryville Road, May 29, 1956, Fell 56-73 (ILL); S. of Cherry Valley, June 6, 1951, Fell & Fell 51-1/9 (ILL).

§ Chordorrhizeae

Rhizomatous perennials with cord-like roots; culms mostly soli¬ tary, with secondary culms axil¬ lary and decumbent at the base; spikes androgynous, head-like, the lowest bracts awn-tipped; styles 2; perigynia spongy, without wings, distinctly nerved, short-beaked; achenes plano-convex.

Only the following species occurs in Illinois.

3. Carex chordorrhiza Ehrh. in L. f. Suppl.

PI. Syst. Veg. 414. 1781. Fig. 3.

Culms slender, subscabrous, canalicu¬ late, 10-35 cm tall, exceeding the leaves, base reclining with the nodes bearing fer¬ tile and sterile culms; leaves 1-4, 2-20 cm long, 1-2 mm broad, canaliculate, ascend¬ ing to appressed with scabrous margins becoming smooth near the base; sheath apex concave, the old sheaths persistent; inflorescence androgynous, 1.00-1.75 cm long, 0. 5-1.0 cm wide; spikelets 3-5, crowded into a head with staminate flow¬ ers conspicuous; pistillate scales ovate, 3-4 mm long, 1.5-2. 5 mm wide, acute to acuminate, reddish-brown with hyaline margins; staminate scales narrowly lan¬ ceolate, 3-4 mm long, 1.0-1. 5 mm wide, reddish-brown to hyaline; bracts (at least the lowest) awn-tipped; perigynia 2-6 per spikelet, 2. 5-3. 5 (-4.0) mm long, 1. 5-2.0 mm wide, plano-convex, margins smooth and thickened, strongly nerved on both sides, glossy, yellow-brown, stipitate, co¬ riaceous and spongy throughout with the beak 0.5 mm long, hyaline-tipped and slightly emarginate dorsally; achene 1.75- 2.00 mm long, 1.25-1.75 mm wide, plano¬ convex, yellow-brown, puncticulate (ex¬ cept margins), continuous with the short style; stigmas 2.

Habitat: Sphagnous swamps.

Range: Newfoundland and eastern Quebec to Alaska, south to Saskatchewan, northern Iowa, northern Illinois, northern Indiana, central New York, southwestern Vermont, and central Maine.

Carex chordorrhiza Ehrh. is best dis¬ tinguished by the reclining base that gives rise to secondary culms in the axils of the previous year’s leaves. The strongly nerved and otherwise distinctive perig¬ ynia provide good identifying characters as well.

Although Fernald (1950) remarks that the perigynia have serrulate beaks, and Mackenzie (1940) illustrates serrations, no Illinois specimens examined exhibited this character.

Evans & Mohlenbrock — Car ex of Illinois

265

Figure 3. Carex chordorrhiza. a. Habit, x 1/4. b. Inflorescence, x 1-1/2. c. Map. d. Staminate scale, x 7-1/2. e. Pistillate scale, x 7-1/2. f. Perigynia, dorsal view, x 7-1/2. g. Perigynia, ventral view, x 7-1/2. h. Achene, x 7-1/2.

266

Transactions Illinois Academy of Science

The roots, lacking in most Illinois col¬ lections, appear wiry and cord-like and can be found at the nodes on some de¬ cumbent bases.

This species is rare in Illinois and may be limited to the boggy areas of the northeastern part of the state as indicated by the four Vasey collections. It appar¬ ently has not been collected in Illinois since the last half of the nineteenth cen¬ tury.

Specimens Examined: LAKE: Without locality, 1862, Vasey s.n. (F 25933); with¬ out locality, 1862, Vasey 29371 (ILL); sphagnous swamps, no date, Vasey s.n.

211538 ); Wauconda, May 30, 1905, Hill s.n. (ILL). MCHENRY: Ringwood, no date, Vasey s.n. (F 25937); Ringwood, no date, Vasey s.n. (ILL). NORTHERN ILLINOIS: No specific location, no date, Vasey 181+08 (ILL).

§ Arenariae

Rhizomatous perennials; culms mostly solitary; spikes androg¬ ynous, distinct or continuous, elon¬ gate, the lowest bract scabrous- awned; styles 2 ; perigynia nerveless ventrally to distinctly nerved on both faces, with a narrow margin or wing, with a serrulate, sharply bi- den tate beak; achenes lenticular or plano-convex.

Key to the Species of § Arenariae in Illinois

1. Rootstock black, fibrillose; culms 50-80 cm tall; inflorescence 2. 5-6. 5 cm long; pistillate scales 2. 5-3. 6 mm long; perig¬ ynia (3-) 10-25 per spikelet, 2. 0-4.1 mm long, narrowly winged to the base; beak 0.75 mm long; achene 1.30-1.75

mm long, without striations . 4. C.

sartwellii

1. Rootstock brown, scaly; culms 6-30 cm tall; inflorescence 1-4 cm long; pistil¬ late scales 3.5-4.75 mm long; perigynia 3-12 per spikelet, 4.50-5.25 mm long, the upper two-thirds narrowly winged; beak 2-3 mm long; achene 1.90-2.25 mm long, finely striate. . . . 5. C. foenea 4. Carex sartwellii Dew. Am. Jour. Sci. 43:90. 1842. Fig. 4.

Plants from thick, fibrous-scaly, hori¬ zontal, black rootstocks; culms subca- naliculate, sharply triangular, scabrous, 5-8 dm tall with the inflorescence well- exceeding the leaves; leaves 3-5, 9-23 cm long, 2.30-4.25 mm broad, arising from the lower half of the culm, ascending to spreading, with the margins scabrous; sheaths often striate ventrally, apex trun¬

cate to concave, some old sheaths per¬ sisting; inflorescence androgynous, 2.5- 6.5 cm long, 0.5-1. 2 cm wide; spikelets 10-35, the upper often entirely staminate and crowded, the lower mostly pistillate and often interrupted; pistillate scales ovate, 2. 5-3. 6 mm long, 1.20-1.75 mm wide, acute to acuminate, reddish-brown, margins hyaline, not concealing the perig¬ ynia; staminate scales narrowly lanceo¬ late, 2. 5-4.0 mm long, 0.5-1. 2 mm wide, acuminate, mostly hyaline throughout; bracts lanceolate, the lowest scabrous- awned; perigynia (3-) 10-25 per spikelet, 2. 0-4.1 mm long, 1.2-1. 6 mm wide, plano¬ convex, substipitate, ovate-lanceolate, distinctly nerved dorsally and ventrally, narrowly winged to the base with the upper 1/3 serrate, light brown, with the bidentate beak 0.75 mm long, serrate and hyaline-tipped; achene 1.30-1.75 mm long, 0. 9-1.0 mm wide, plano-convex, reddish- brown, puncticulate, margins single- nerved, jointed to a short style; stigmas 2.

Habitat: Low wet prairies and mead¬ ows, creek and river bottoms, marshes, dunes, peaty swamps, and open cold bogs.

Range: Southwest Quebec to northern British Columbia, south to Colorado, Ne¬ braska, Missouri, Illinois, Indiana, Ohio, and western New York.

Boott (1865) , apparently seeing no char¬ acters to distinguish Carex sartwellii Dew. from European Carex disticha Huds., illus¬ trates both as C. disticha. However, Boott’s figures 2a, b of perigynia and achenes from French material are con¬ siderably different from those of C. sart¬ wellii. We feel the difference is enough to consider both as distinct species.

Carex disticha Huds. is not accurately reported from Illinois. The Illinois collec¬ tions reported to be C. disticha are actu¬ ally C. sartwellii. These are noted in the list of specimens examined for this species.

Carex sartwellii has an extremely vari¬ able inflorescence. The spikelets of the linear to ellipsoid-cylindric head vary from very crowded to much interrupted. The upper half of the inflorescence is often entirely staminate ( Bebb s.n. [E 32311]) with the spikelets much smaller and more sharply pointed and hyaline than the lower, subglobose, pistillate ones. In other material, pistillate spikelets occur throughout the inflorescence with a few staminate scales occurring in the tips of the spikelets. Further still, some speci¬ mens ( Wolf s.n. [F 211662] and Fell 533098 [ISM]) have spikelets aggregated into a short, compact head.

Standley and Steyermark have deter¬ mined their collection 28138 (F) as C. sartwellii Dew. var. stenorrhynca Her¬ mann. However, the perigynia length of

Evans & Mohlenbrock — Car ex of Illinois

267

Figure 4. Car ex sartwellii. a. Habit, x 1/4. b. Inflorescence, x 1. c. Map. d. In¬ florescence, x 1. e. Staminate scale, x 7-1/2. f. Pistillate scale, x 7-1/2. g. Perigynia, ventral view, x 7-1/2. h. Perigynia, dorsal view, x 7-1/2. i. Achene, x 7-1/2.

268

Transactions Illinois Academy of Science

this material falls within the variable limits of typical C. sartwellii, not 4. 0-4. 5 mm long as Hermann (1938) describes in the variety stenorrhynca. Thus this va¬ riety should be excluded from Illinois.

The continuous black rootstock com¬ bined with scabrous culms and distinctly nerved perigynia provide good identifying characters.

Although Fernald (1950) remarks that the pistillate scales are obtuse or mucro- nate, all Illinois material examined and Mackenzie’s illustrations show the scales to be acute to acuminate. Also, the green- striate sheath mentioned by Fernald and Mackenzie, although occurring often, is not present in all Illinois material ( Dobbs s.n. [ILLS 18511, 10030]).

Carex sartwellii, a common sedge in the northern United States and southern Canada, is found only occasionally in the bottoms, swamps, and bogs of the north¬ ern third of the state. However, it does occur as far south as Fulton and Menard counties. The Jones and Fuller (1950) re¬ port of its occurrence in Will County is not verified by this study.

Specimens Examined: COOK: Ash- burn, May 26, 1906, Cowles 8172-0 (ILLS); northeast of Bartlett, May 21, 1956, Bennett s.n. ( ILLS 69232); east side of Calumet Lake, June 2, 1948, Steyer- mark 68221 (F, ILL); Chicago, no date, Monroe s.n. (ff 68997 ) [originally deter¬ mined C. disticha Huds.]; Chicago, June, 1891, McDonald s.n. (F 398575 [originally determined C. disticha Huds.]; near Cole- hour, April 29, 1876, Hill 122 (ILL); near Chicago, June 5, 1890, Moffatt 156 (ILL); near Chicago, June 7, 1890, McDonald s.n. (ILL). DEKALB: W. of Kirkland, June 7, 1961, Evers 69180 (ILLS); north of Fairdale, June 1, 1953, Fell 53276 (ISM, SIU) ; near Kirkland, June 10, 1953, Fell 53396 (ISM), 53398 (ISM). DUPAGE: N. of Wheaton, May 22, 1894, Moffatt 91 \ (ILL); N. of Wheaton, May 2, 1894, Mof¬ fatt 152 (ILL). FULTON: Canton, no date, Wolf s.n. (F 211662), (F 211663 ); without location, no date, Brendel s.n. (ILL) [originally determined C. disticha Huds.]. HENRY: E. of Geneseo, May 30, 1937, Dobbs s.n. ( ILLS 10030). KAN¬ KAKEE: Kankakee, May 17, 1870, Hill s.n. 206J)65) [originally determined C. disticha Huds.]; Kankakee, May 29, 1870, Hill s.n. (ILL) [originally determined C. disticha Huds.]. LAKE: Waukegan dunes, June 13, 1940, Standley & tSteyermark 28138 (F) ; Waukegan, June 5, 1916, Benke 1512 (F) [originally determined C. siccata Dew.]. LEE: Near Amboy, June 8, 1959, Long 939 (ILL). MACON: Decatur, May 22, 1899, Clokey 31235 (ILL). MENARD:

Athens, 1861, Hall s.n. (F 32212), (F 55050). MCHENRY : Ringwood, no date, Vasey s.n. (ILL). OGLE: N. of Davis Junction, May 24, 1953, Fell 53396 (ISM), 53398 (ISM). PEORIA: Peoria, no date, Brendel s.n. (ILL). STEPHENSON: Northeast of Ridott, June 28, 1953, Fell 53202 (ISM), 53-203 (SIU). WINNE¬ BAGO: Near Rockford, May 28, 1952, Fell 52-126 (ILLS), 52-136 (ISM); Foun- taindale, 1870, Bebb s.n. (F 32331) [origi¬ nally determined C. disticha Huds.]; Rockton Township, June 10, 1952, Fell 52306 (ISM) ; N. of Shirland, June 8, 1952, Fell 52268 (ISM); Kent Creek, July 1, 1951, Fell 51251 (ISM); North Central Avenue, Rockford, June 14, 1952, Fell 52356 (ISM, SIU). WITHOUT PRE¬ CISE LOCALITY: No date, Vasey s.n. (F 32751, 32752, 32753) [F 32752 appar¬ ently cited in Boott as C. disticha Huds.]; no date, Vasey 18525 (ILL); June, 1860, Hall s.n. (F 206219) [originally deter¬ mined C. disticha Huds.].

5. Carex foenea Willd. Enum. Hort. Berol.

2:957. 1809. Fig. 5.

Carex siccata Dew. Am. Journ. Sci. 10:278.

1826.

Roots from brown, scaly, slender, hori¬ zontal rootstock; culms slender, mostly fertile, 15-75 cm tall, sharply angled and scabrous at the apex, becoming rounded, canaliculate and smooth near the base, equalling or exceeding the leaves; leaves 3-5, 6-30 cm long, 2-3 mm broad, flat¬ tened, becoming triangular at the tip, as¬ cending to spreading with the margins scabrous; sheath apex truncate to con¬ cave; inflorescence androgynous, 1-4 cm long, 0. 5-1.0 cm broad; spikelets 4-8, slightly interrupted to widely spaced at the base, becoming indistinguishable at the apex, the lower 1-3 spikelets usually pistillate, the middle spikelets staminate and the terminal gynecandrous and larger; pistillate scales lanceolate, 3.50-4.75 mm long, 1. 5-2.0 mm broad, acute to mostly acuminate, light reddish-brown with hya¬ line margins, shorter than the perigynia; staminate scales acuminate, 3. 5-4. 2 mm long, 1.0-1. 5 mm broad, light brown to hyaline; bracts lanceolate, the lowest en¬ circling the culm with scabrous awns; perigynia 3-12 per spikelet, 4.50-5.25 mm long, 1.5-1. 8 mm broad, plano-convex, cuneate, substipitate, ovate-lanceolate, light reddish-brown, coriaceous, distinctly nerved on both faces, rarely nerveless ventrally, the upper 2/3 narrowly winged and serrulate with the beak 2-3 mm long, serrulate, sharply bidentate and obliquely cleft dorsally; achene 1.90-2.25 mm long, 1.25-1.50 mm broad, lenticular, substi¬ pitate, yellow-brown, finely striate, punc- ticulate, jointed to the style; stigmas 2.

Evans & Mohlenbrock — Carex of Illinois

269

Figure 5. Carex foenea var. foenea. a. Habit, x 1/2. b. Inflorescence, x 1. c. Map. d. Inflorescence, x 1. e. Inflorescence, x 1-1/2. f. Staminate scale, x 7-1/2. g. Pistillate scale, x 7-1/2. h. Perigynia, dorsal view, x 7-1/2. i. Perigynia, ventral view, x 7-1/2. j. Achene, x 7-1/2.

270

Transactions Illinois Academy of Science

Habitat: Mostly prairie or sandy soil, sometimes sandy woods and roadsides.

Range: Southwest Quebec to Mac¬ kenzie, south to Arizona, New Mexico, Illinois, northern Indiana, Ohio, New Jersey, and New York.

Although this species has been called Carex siccata, Svenson (1938) has given reasons for accepting C. foenea as the proper binomial.

Two varieties may be recognized.

1. Perigynia nerved on both faces, taper¬ ing gradually to the beak .

. 5a. C. foenea var. foenea

1. Perigynia nerveless on the ventral face,

tapering abruptly to the beak .

. 5b. C. foenea var. enervis

5a. Carex foenea Willd. var. foenea

Perigynia nerved on both faces, taper ing gradually to the beak.

Carex foenea Willd. var. foenea strongly resembles Carex praegracilis Dew. because of the more slender inflorescence. How¬ ever, C. foenea is distinguished by its pe¬ rigynia which are many-nerved on both faces and are larger with a proportion¬ ately longer beak, by its rhizomes brown and scaly, and by its middle spikelets usually wholly staminate.

Some variation can be found in this taxon. Fell 211 (SIU) contains one culm bearing perigynia with widely divergent teeth while the other characters are typi¬ cal.

A few collections have been erroneously identified as this taxon. These are Benke 3591 (F) from Elgin in Kane County, pre¬ viously determined as C. siccata Dew., which actually is C. praegracilis Dew., and Cowles 8007-0 (ISM) from South Chicago, Cook County, determined as C. siccata Dew., which actually is C. bebbii Olney.

Specimens Examined: BOONE: North¬ east of Belvidere, May 10, 1954, Fell 55215 (ISM). COOK: Lakeview, May 15, 1884, Ohlendorf s.n. (F 1358087). DE¬ KALB: Near Fairdale, May 20, 1953, Fell 53112 (ISM). MENARD: Without specific locality, 1876, Hall s.n. (F 206255) ; Athens, 1861, Hall s.n. (F 32213). MCHENRY: Ringwood, no date, Vasey s.n. (JF 327 f 5, F 325752). STEPHENSON: Northeast of Ridott, May 28, 1953, Fell 53211 (ISM, SIU). WINNEBAGO: E. of Rockford, June 5, 1952, Fell 52-228 (ILLS), 52-227 (ISM); S. of Rockford, May 20, 1951, Fell 5153 (ISM) ; S. of Rock Cut, June 23, 1952, Fell 52573 (ISM); River Road S. of Cherry Valley, May 31, 1952, Fell 52176 (ISM); Sugar Creek Forest Preserve, May 18, 1952, Fell 5286 (ISM) ; Sugar Creek Forest Preserve, May 19, 1951, Fell 5150 (ISM). WITHOUT

PRECISE LOCALITY: 1896, Dewey s.n.

(F 567223).

5b. Carex foenea Willd. var. enervis Evans & Mohlenbr., var. nov. Fig. 6. Perigynium enerve ventrali superficie, rostro abrupte decrescenti.

Type: French s.n. (SIU), from Illinois.

The type collection was made in 1869 by G. H. French in “sandy plains Ill.” Although Irvington, Illinois [Washington County] is written on the label, the col¬ lection is probably not from there since Washington County seemingly is a little too far south. French, who lived for a while in Irvington, is known to have written Irvington on the labels of other specimens which were actually collected elsewhere in Illinois.

The lack of nerves on the ventral face of the perigynia and the more abruptly tapering beak differentiate this taxon from C. foenea var. foenea.

Index to Exsiccatae

Bebb, M. S. s.n. (4).

Benke, H. C. 1512 (4), 3591 (2).

Bennett, H. R. s.n. (4).

Brendel, F. s.n. (4).

Clokey, I. W. 31235 (4).

Cowles, H. C. 8172-0 (4).

Dewey, _ s.n. (5a).

Dobbs, R. H. s.n. (4).

Evers, R. A. 69180 (4).

Fell, E. W. 5140 (5a), 5153 (5a), 51251 (4), 5286 (5a), 52105 (21, 52126 (4),

52136 (4), 52176 (5a), 52-227 (5a), 52- 228 (5a), 52268 (4), 52306 (4), 52356 (4), 52473 (5a), 53112 (5a), 53150 (4), 53202 (4), 53-203 (4), 53211 (5a), 53276 (4), 53371 (2), 53396 (4), 53398 (4),

53856 (2), 54214 (5a), 55-141 (2), 55-318

(2), 55-440 (2), 56-73 (2), 56-91 (2),

56-113 (2), 57-9 (1), 57-68 (1), 57-157

(1) , 57-248 (1).

Fell, E. W. & Fell, G. B. 51-149 (2).

Fell, E. W. & Fuller, G. D. 17200 (2). French, G. H. s.n. (5b).

Hall, E. s.n. (4, 5a).

Hill, E. J. s.n. (3, 4), 122 (4).

Long, _ 939 (4).

McDonald, F. E. s.n. (4).

Moffatt, W. S. 94 (4), 142 (4), 156 (4). Munroe, H. F. s.n. (4).

Ohlendorf, _ s.n. (5a).

Spongberg, S. 61/38 (2), 61/38 A (2), 61/41

(2) , 61/41 A (2).

Standley, P. C. & Steyermark, J. A. 28138 (4).

Steyermark, J. A. 68221 (4).

Swink, F. s.n. (4).

Vasey, G. s.n. (3, 4, 5a), 18408 (3), 18425 (4), 29311 (3).

Wolf, J. s.n. (4).

Evans & Mohlenbrock — Car ex of Illinois

271

Figure 6. Carex foenea var. enervis. a. Habit, x 1/2. b. Inflorescence, x 2. c. Map. d. Staminate scale, x 7-1/2. e. Pistillate scale, x 7-1/2. f. Perigynia, dorsal view, x 7-1/2. g. Perigynia, ventral view, x 7. h. Achene, x 15.

272

Transactions Illinois Academy of Science

Literature Cited

Boott, F. 1858. Illustrations of the Genus Carex. William Pamplin, London. Fell, E. W. 1958. New Illinois Carex Rec¬ ords. Rhodora 60:115-116.

Fernald, M. L. 1950. Gray’s Manual of Botany. 8th ed. The American Book Company, New York. 1632 pp. Hanson, H. C. and E. D. Churchill. 1961. The Plant Community. Reinhold Publishing Company, New York. 218

pp.

Hermann, F. J. 1938. New and Otherwise Interesting Plants from Indiana. Rho¬ dora 40:77-86.

Jones, G. N. and G. D. Fuller. 1955.

Vascular Plants of Illinois. The Uni¬ versity of Illinois Press, Urbana, and The Illinois State Museum, Spring- field. 593 pp.

Mackenzie, K. K. 1940. North American Cariceae. The New York Botanical Gardens, New York. 547 pp.

Svenson, H. K. 1938. Carex foenea, C. straminea, and C. albicans in Willden- ow’s Herbarium. Rhodora 40:325-331.

Weaver, J. E. and F. W. Albertson. 1956. Grasslands of the Great Plains. Johnsen Publishing Company, Lincoln, Nebraska. 395 pp.

Manuscript received June 21+, 1970

GASTRIC MORPHOLOGY IN SELECTED MORMOOPID AND GLOSSOPHAGINE BATS AS RELATED TO

SYSTEMATIC PROBLEMS

G. LAWRENCE FORMAN

Department of Biology, Rockford College, Rockford, Illinois 61101

Abstract. — Stomachs of selected spe¬ cies of two groups of North American bats [Mormoopidae and Glossophaginae (Phyl- lostomatidae)] were studied grossly, his¬ tologically, and histochemically. The stomachs of two mormoopids, Pteronotus parnellii and Mormoops megalophylla are most similar in gross and histological fea¬ tures to those of fish-eating bats of the family Noctilionidae. Species of the pre¬ dominantly nectar-feeding Glossopha¬ ginae have stomachs of two basic mor¬ phological forms, which are considered to probably reflect early divergence in food habits. Additionally, studies of the histo¬ chemistry of gastrointestinal mucins sug¬ gest a similar dichotomy in glossophagines. The above results are reviewed in light of several recent studies of the systematics of the Mormoopidae and Glossophaginae. It is concluded that a comprehensive study of the food habits of glossophagines should be undertaken to test some pro¬ posed relationships between gastric mor¬ phology and feeding characteristics in these bats.

Recent investigations of certain genera and subfamilies of the North American leaf-nosed bats, family Phyllostomatidae, have proposed noteworthy changes in the tradi¬ tional systematic arrangements, and explanations of phylogeny of these groups. Insect-feeding bats of the subfamily Chilonycterinae have been reviewed critically by Smith (1971) who regards them as distinc¬ tive enough to be relegated to a separate family, Mormoopidae. Vaughn and Bateman (1970) sup¬ port this proposal with evidence from studies of the functional mor¬ phology of flight. Phillips (1971) examined the deciduous and adult dentitions of selected nectar-feed¬ ing species of the subfamily Glosso¬ phaginae. He concluded that this group is either diphylletic in origin and not a natural assemblage, or

that the subfamily contains a di¬ chotomy of types that is the prod¬ uct of an early evolutionary diver¬ gence. Based on karyotypic evi¬ dence, Baker (1967) previously had reported a dichotomy in the glosso¬ phagines, but the two species groups devised by him do not correspond precisely with those of Phillips.

In an effort to further examine these three systematic proposals, bats of species representing the groups were selected and their stom¬ achs were examined morphologi¬ cally and compared with those of closely related taxa. The relation¬ ship between the Mormoopidae and the fish-eating bats of the family Noctilionidae also was studied and the results are discussed herein. The systematic value of such studies of gastrointestinal morpholopy has been discussed previously by For¬ man (1971) and Schultz (1970).

Materials and Methods

Preserved stomachs of one spe¬ cies of mormoopid and four species of glossophagines were examined grossly, histologically, and histo¬ chemically. The kinds and numbers examined were as follows: Mor¬ moopidae; Mormoops megalophylla (2), Glossophaginae; Glossophaga commissarisi (2), Anoura geoffroy (1), Choeroniscus godmani (1) and Lichonycteris obscura (2). Previous accounts of gastric morphology and histochemistry in the mormoopid Pteronotus parnellii (Forman, 1971), and the glossophagines Glossophaga soricina (Forman, op. cit.) and Lep- tonycteris sanborni (Rouk and Glass, 1970) are employed in discussions to follow.

273

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Transactions Illinois Academy of Science

All specimens from which stom¬ achs were removed are stored in the Museum of Natural History, The University of Kansas.

Standard histological and histo- chemical procedures employing Harris’ haematoxylin and eosin, and periodic acid Schiff-Alcian blue for gastric mucus were used (see For¬ man, 1971). Hale’s colloidal iron reaction was used in addition to Alcian blue as a test for acid muco¬ polysaccharides (procedure after Lillie, 1965). All stomachs were sectioned at five to seven microns. Drawings of stomachs were pro¬ duced by projecting midlongitudi¬ nal sections onto paper, by means of a photographic enlarger.

Results

Gastric Morphology in the Mor- moopidae. The stomach of Mor- moops (Fig. 1) is simple in overall configuration, and is nearly identi¬ cal in topography to that of Ptero- notus parnellii (Forman, 1971). The stomach is symmetrical (gastroeso¬ phageal junction midway along les-

Figure 1. Mid-longitudinal represen¬ tation of the stomach of Mormoops mega- lophylla. Explanation of symbols: CG, cardiac glands; BG, Brunner’s glands; PG, pyloric glands; TZ, transitional zone; FG, fundic glands; IC, incisura cardiaca; CV, cardiac vestibule; PS, pyloric sphinc¬ ter.

ser curvature) with a short, pointed fundic caecum, and an equally short and unusually broad pyloric end- piece. The cardiac vestibule is min¬ ute, as it is in nearly all other in¬ sectivorous bats examined thus far (see Forman, 1971).

The pyloric sphincter is asym¬ metrical, as in Pteronotus, the valve on the greater curvature being longer and narrower than that of the lesser curvature. However, the marked reduction in mass of the lesser valve seen in Pteronotus is lacking in Mormoops. The circular muscle layer of the stomach wall is extremely thick, as in Pteronotus.

The glands of Brunner at the gas¬ troesophageal junction of Mormoops are indistinguishable in cellular morphology from those of Pterono¬ tus parnellii, as well as Noctilio le- porinus and N. Labialis (Nocti- lionidae). The tubules are narrow and the cells are small with ex¬ tremely small, flattened nuclei not juxtaposed to the basement mem¬ brane. This condition is in contrast to the broader tubules with larger, circular juxtaposed nuclei usually found in Brunner’s glands in bats of the families Vespertilionidae, Phyl- lostomatidae, Emballonuridae and Molossidae. Brunner’s glands are unusually abundant in Mormoops (Fig. 1) and Pteronotus, a condition shared with Noctilio. The extreme breadth of the duodenum at the gastroesophageal junction of Mor¬ moops, Pteronotus, and Noctilio is, in part, due to the unusually large complement of Brunner’s glands in these genera.

The distribution of gastric mu¬ cosa in Mormoops closely resembles that of Pteronotus, and thus is dis¬ tinctive among bats studied to date (except for Noctilio ) by virtue of having an extremely narrow zone of mucus-producing pyloric glands (Fig. 1). The proportion of gastric mucosa that may be considered

Forman — Bat Gastric Morphology

275

transitional (between fundic and pyloric glands) is extremely high in mormoopids, although a similar condition is found to occur in some phyllostomatids and in Noctilio. The relative frequency of mucosal folds is low in Mormoops and Pte- ronotus, considerably lower than in any other kinds thus far examined.

The fundic mucosa of Mormoops, which produces hydrochloric acid and pepsin, has a number of fea¬ tures found to occur additionally only in Pteronotus and in the Noc- tilionidae. The fundic tubules are long, highly convoluted, and ex¬ tremely narrow (Fig. 2) in contrast to the broader, often shorter glands

in members of other families of North American bats (Fig. 3). All cellular elements of the fundic glands (parietal, mucous neck, chief, and argentaffin cells) are extremely small in comparison to other bats, and correspondingly are relatively abundant. In comparison with other species, nuclei of these cells are of moderate size, suggesting a decrease in cytoplasmic mass. Zymogenic (chief) cells are abundant within the bases of fundic glands only in the cardiac vestibule and lesser cur¬ vature of Mormoops and Pterono¬ tus.

Gastric Morphology in the Glosso- phaginae. Careful examination of

Figure 2. Fundic glands in the stom- Figure 3. Fundic glands in the stom¬ ach of Mormoops megalophylla. Note the ach of Glossophaga soricina (family Phyl- great length and narrowness of these lostomatidae). Note the greater breadth glands (X200). of these glands as compared to those in

Fig. 2 (X200).

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Transactions Illinois Academy of Science

Figures 4 to < reveals the extensive variation in gross morphology of stomachs within the nectar-feeding Glossophaginae. Stomachs of glos- sophagines thus far examined can

2 mm

Figure 4. Mid-longitudinal represen¬ tation of the stomach of Glossophciga com- missarisi. For explanation of symbols, see Fig. 1.

Figure 5. Mid-longitudinal represen¬ tation of the stomach of Anoura geoffroy. For explanation of symbols, see Fig. 1.

Figure 6. Mid-longitudinal represen¬ tation of the stomach of Choeroniscus god- mani. For explanation of symbols, see

Fig. 1.

Figure 7. Mid-longitudinal represen¬ tation of the stomach of Lichonycteris obscura. For explanation of symbols, see Fig. 1.

Forman — Bat Gastric Morphology

277

be divided into two groups on the basis of topographic features and various features of the gastric mu¬ cosa. Group A includes Glossophaga soricina (examined by Forman, 1971), G. commissarisi (Fig. 4 ),Lep- tonycteris sanborni (examined by Rouk and Glass, 1970), and Anoura geoffroy (Fig. 5). Group B includes Choeroniscus godmani (Fig. 6) and Lichonycteris obscura (Fig. 7).

The stomachs of Group A are characterized by a relatively long, narrow fundic caecum, and a com¬ paratively small or no cardiac ves¬ tibule (tubular portion of stomach between gastroesophageal junction and lesser curvature). The incisura cardiaca (fold in the stomach wall at the junction of the cardiac ves¬ tibule and fundic caecum) is mod¬ erately to extensively developed. The terminal portion of the stom¬ ach (tubular segment between the gastroesophageal junction and py¬ loric sphincter) is always moder¬ ately well developed. The pyloric sphincter possesses comparatively short valves, with well developed musculature, and is always assym- metrical, with the valve of the greater curvature being the most massive.

In regard to gross morphology of the stomach, Glossophaga soricina is most deviant from the general plan within Group A. A cardiac vestibule is completely lacking in this species, and the apparent re¬ lationship between this condition and insect-feeding in this bat prev¬ iously has been discussed (Forman, 1971).

The stomachs of Group B are distinguished from those of Group A on the basis of morphology of the fundic caecum and pyloric sphincter. The fundic caeca of Choeroniscus and Lichonycteris are short, rounded, and slightly dilated on the dorsal or “back’' surface. An incisura cardiaca is completely lack¬

ing. The cardiac vestibule is gen¬ erally better developed than those of Group A, and more closely ap¬ proximates the type found to occur in fruit-eating leaf-nosed bats. The terminal portion of the stomach is relatively elongate as in Group A, however, the pyloric sphincter dif¬ fers by having narrower valves of greater length. This latter condi¬ tion more closely resembles the pat¬ tern found in frugivorous phyllo- stomatids, than that of Group A.

The mucus-producing glands of Brunner reveal distributional and structural differences between the two groups. These submucosal glands within the duodenal wall at the pyloric sphincter are relatively more abundant in both species of Glossophaga and in Anoura than in Choeroniscus and Lichonycteris. They are limited to a narrow band on only the lateral one-half of the py¬ loric sphincter in Lichonycteris (Fig. 7). The tubules of Brunner’s glands are more highly coiled and consider¬ ably broader in Group A than in the second group, suggesting greater production of mucus from Brun¬ ner’s glands in Glossophaga and An¬ oura. Of the three species examined in Group A, Anoura has Brunner’s glands which most closely approach those of Group B in morphology of the tubules.

The entire inner lining of the stomach of all glossphagines is oc¬ cupied by typical tubular gastric glands. Differences in the distribu¬ tion of various portions of the gas¬ tric mucosa within the Glossopha- ginae are not striking, except for

the extremelv limited distribution */

of true fundic glands in Choeronis¬ cus godmani (Fig. 6). The zone of transitional mucosa between fun¬ dic and pyloric glands is character¬ istically broad in glossophagines, being maximal in Choeroniscus with the absence of zymogenic cells

278

Transactions Illinois Academy of Science

throughout the cardiac vestibule (Fig. 6).

Although distinctive differences in distributions of glandular types are not apparent among the glos- sophagines, two conditions of the frequency of zymogenic cells are found to occur within the fundic glands. Fundic glands of Glosso¬ phaga and Anoura contain relatively large numbers of zymogenic cells within the basal one-fourth to one- third of each tubule. These cells are abundant throughout that por¬ tion of the stomach in which they occur, being most abundant within the fundic caecum (Anoura) and lesser curvature (Glossophaga) . In contrast, chief cells are consider¬ ably less frequent in both Lichonyc- teris and Choeroniscus , being least abundant in the latter, and are gen¬ erally restricted to the lower one- seventh to one-tenth of each tubule in both. Parietal cells, which oc¬ cupy only the midregion of fundic tubules in Glossophaga and Anoura, are usually found among chief cells in Lichonycteris, and are always to¬ gether with these cells in Choeronis¬ cus . The most abundant concentra¬ tion of chief cells in both species of Group B is within the extreme apex of the fundic caecum.

Histochemistry of the Gastric Mu¬ cosa. Staining of stomachs with Al- cian blue, Hale’s collidal iron stain, and the periodic acid Schiff reaction (paS) revealed few differences in the quality of gastric mucus among mormoopid or glossophagine bats. Alcian blue and Hale’s iron, both of which are considered to color acid mucopolysaccharides, stained mu¬ cus only of the cardiac glands in Mormoops and Pteronotus. Mucus of the surface epithelium of these genera stained strongly with paS (for neutral mucins), as did cells of the gastric pits within the cardiac and pyloric glands. Mucous neck cells interspersed among parietal

cells in the fundic and transitional glands stained moderately strong with paS, as did cells within the basalmost one-fifth of the pyloric glands. Brunner’s glands gave a strong reaction to paS in both spe¬ cies of mormoopids. With the ex¬ ception of somewhat less reactivity in the mucous neck cells of Mor¬ moops, there was little difference between mormoopids in staining with paS.

The surface epithelium of all glos¬ sophagine stomachs gave a strong reaction to paS, but none with Al¬ cian blue or Hale’s colloidal iron. Results also were consistent for the cardiac glands, with the upper three- fourths of each tubule staining strongly with paS and not at all with either acid mucopolysaccha¬ ride technique. The basal one-fourth of the cardiac gland tubules stained only moderately with paS, and moderate to heavily with both Alcian blue and Hale’s colloidal iron.

The pyloric glands and mucous neck cells of glossophagines did not stain with Hale’s iron, but revealed interspecific variation in staining with Alcian blue. Mucous neck cells within the lower portions of the fundic glands stained strongly in Glossophaga, moderately in Anoura, and not at all in Choeroniscus and Lichonycteris.

The pyloric glands stained in a similar fashion. Those of Glossopha¬ ga (both species examined) colored intensely throughout the length of the tubule, whereas, those of An¬ oura stained with equal intensity, but only within the basal arc of each tubule. The pyloric glands of Choeroniscus and Lichonycteris did not stain with Alcian blue.

Discussion

Revieiv of Food Habits. The food of mormoopid bats is fairly well es¬ tablished in the literature as con¬ sisting mostly of flying insects. In

Forman — Bat Gastric Morphology

279

contrast, however, the food habits of many glossophagine species, which previously were assumed to be obligate nectar feeders, are in¬ adequately or inaccurately docu¬ mented. Glossophaga soricina now may be considered an omnivore (see Goodwin and Greenhall, 1961; Ara- ta et al., 1967), that consumes in¬ sects as well as nectar. The food of Anoura geoffroy is said by Goodwin and Greenhall (op cit.) to consist of nectar and soft fruit pulp. There is little evidence, as yet, that this bat takes insects. Choeroniscus godmani is assumed to be an obligate nectar feeder, but it is of interest to note that C. intermedins on Trinidad has been reported to include numerous insects in its diet (based on a single specimen examined by Goodwin and Greenhall, op cit.). The teeth of Choeroniscus are greatly reduced in breadth and height in compari¬ son to most other glossophagines. Thorough masceration of insect ma¬ terial, which is characteristic of in¬ sect-feeding bats, seems unlikely in Choeroniscus, and thus insect feed¬ ing by this bat is probably rare. Stomach contents of Leptonycteris sanborni, as examined by Hoff- meister and Goodpaster (1954), con¬ sisted of 92 per cent pollen and eight per cent insect remains. Al¬ though Barbour and Davis (1969) reported that this species will eat a variety of fruits in captivity, in¬ sects possibly are not uncommon in the diet of this species. Leptonycteris nivalis is not formally known to take insects, however, it seems likely that general accounts of Leptonyc¬ teris sanborni apply to this species. Little is known, directly, of the food habits of Choeronycteris mexi- cana. Indirect evidence (heads and faces of specimens covered with pollen, as reported by Barbour and Davis, op cit.) would suggest that this bat probably is an obligate nectivore. Little information is

available concerning the food habits of Lichonycteris obscura, but it is as¬ sumed that this species feeds on pollen and nectar.

Gastric Morphology in Relation to Feeding. Numerous morphological features of the stomachs of Pte- ronotus and Mormoops (simple over¬ all configuration, thick and assym- metrical pyloric sphincter, small fundic caecum, and an extensive complement of zymogenic cells) are strikingly similar to conditions found in insect-feeding bats of the large North American family Ves- pertilionidae. Elongation of the ter¬ minal portion of the stomach, found to occur in North American insec¬ tivorous bats of the families Molos- sidae, Natalidae, and Emballonuri- dae, as well as in frugivorous and nectivorous phyllostomatids, is lacking in mormoopids. Mormoop- ids thus appear to parallel only ves- pertilionids in maintenance of a generalized gastric form. The glands of Brunner at the gastroesophageal junction are extremely abundant in both Pteronotus and Mormoops, resembling the condition found in other insectivorous, as well as fish¬ eating kinds. Non-carnivorous bats generally possess relatively fewer and less well developed Brunner’s glands than carnivorous kinds.

Upon a review of the morpho¬ logical features of stomachs of the glossophagines examined, it can be seen that there are two general structural forms. Of genera exam¬ ined, Glossophaga and Anoura have stomachs of one form; Choeroniscus and Lichonycteris, the other. The somewhat more saccular stomach with expanded cardiac vestibule found in Choeroniscus and Lichonyc¬ teris more closely approximates the form found in frugivorous phyllo¬ stomatids, than does the form found in the other group of glossophag¬ ines. The stomachs of Glossophaga and Anoura are more tubular, with

280

Transactions Illinois Academy of Science

less well developed cardiac ampul¬ lae, more closely resembling gastric configurations of insectivorous or carnivorous species. In the absence of adequate studies of food habits of most glossophagine genera, it would be premature to make an unqualified statement about gas¬ tric morphology as it relates to feed¬ ing in the Glossophaginae. How¬ ever, there is sufficient evidence to suggest that there are two morpho¬ logical forms of stomachs, and that an increase in stomach complexity is probably related to a decrease in insect- or flesh-eating. The descrip¬ tive account of gastric morphology, as well as an included diagram, of the stomach of Leptonycteris san- borni by Rouk and Glass (1970) closely parallels that of Glossophaga soricina (Forman, 1971), G. Com- missarisi, and Anoura geoffroy. For this reason, the stomach of Lepto¬ nycteris sanborni here is considered to be most like those of Group A.

Several microanatomical fea¬ tures, which are found in two con¬ ditions or states in glossophagines, apparently are related to food hab¬ its, and reflect trends found to oc¬ cur in other groups of bats. For ex¬ ample, there are two patterns of distribution of the glands of Brun¬ ner in the Glossophaginae that tend to parallel the two conditions ob¬ served in frugivorous and insectiv¬ orous or carnivorous species. Infre¬ quent Brunner’s glands, as found in Choeroniscus and Lichonycteris, is common, although not universal among frugivores. On the other hand, relatively abundant glands (' Glossophaga and Anoura ) is a char¬ acteristic of carnivorous bats. Ad¬ ditionally, zymogen or ‘Thief” cells (those cells that presumably secrete proteolytic enzymes) are abundant in the fundic mucosa only in Glos¬ sophaga and Anoura. This probably is a further indication of the car¬ nivorous habits of these bats.

Studies of gastrointestinal histo¬ chemistry revealed no striking dif¬ ferences between insect- and nec¬ tar-feeding bats. The significance of acid mucopolysaccharide in the fundic and pyloric glands of Glosso¬ phaga and Anoura, and the appar¬ ent absence of this material, except for universal staining in the cardiac glands, in either of the other two glossophagines or in the insect-feed¬ ing mormoopids remains unex¬ plained.

Systematic Relationships as Re¬ vealed by Gastric Morphology. Sys¬ tematic relationships among vari¬ ous closely related families of North American bats (Phyllostomatidae, Mormoopidae, Noctilionidae and Emballonuridae) recently have been reviewed by Smith (1971). He stated, on the basis of several fea¬ tures of external and internal mor¬ phology, that the Mormoopidae, Noctilionidae and Phyllostomatidae resemble one another more than they resemble any other New World group. He further suggested, on the basis of morphology of the male phallus and position of wing attach¬ ment, that mormoopids and noc- tilionids may be more closely re¬ lated than previously thought. An examination of gastric structure in these two groups provides support for Smith’s proposal.

Forman (1971) studied gastric morphology in the insect-feeding Noctilio labialis, and reported the presence of several features, which are noted here as being found to¬ gether also only in Mormoops and Pteronotus. A large complement of Brunner’s glands occurs at the gas¬ troesophageal junction of N. la¬ bialis. These glands are indistin¬ guishable in cellular morphology from those of the mormoopids ex¬ amined, and differ from most other bats in possessing small cells with extremely small nuclei. Stomachs of Noctilio and the Mormoopidae

Forman — Bat Gastric Morphology

281

also resemble one another in having extremely limited distributions of pyloric glands, extremely long, nar¬ row, and slightly coiled, gastric glands, short and pointed fundic caeca, and a relatively short ter¬ minal portion (that portion between the gastroesophageal junction and pyloric sphincter). None of the afore-mentioned features has yet been found in species of the Phyl- lostomatidae. A suggestion that si¬ milarity in gastric morphology is entirely attributable to consump¬ tion of similar foods by noctilionids and mormoopids, and therefore of little or no taxonomic value, would not be in order. Most of the mor¬ phological features discussed above have not been found to occur in other families of bats that charac¬ teristically consume insects or flesh (e.g. Natalidae, Vespertilionidae and Molossidae).

Baker (1967) studied several spe¬ cies of Glossophagines and found three species-groups based on chro¬ mosome number and morphology. One group included Choeronycteris mexicana and Choeroniscus godmani. The second group was composed of Leptonycteris sanborni and three spe¬ cies of Glossophaga (plus three gen¬ era of phyllostomatids not assigned to the subfamily Glossophaginae) . The third group included only An- oura geoffroyi. Baker suggested that Leptonycteris and the species of Glos¬ sophaga form a natural assemblage because their karyotypes are quite different from those of other phyl¬ lostomatids in that karyotypic grouping. Differences between the first and second groups included differences in both fundamental number and morphology of indi¬ vidual chromosomes. Additional support for the distinctiveness of karyotypic groups in the Glosso¬ phaginae is provided in the obser¬ vation by Baker that similarities between karyotypes of Phyllosto-

mus (Phyllostomatinae) and Lepto¬ nycteris are greater than those be¬ tween the Choeroniscus group and Leptonycteris. The karyotype of An- oura was said to be somewhat in¬ termediate in having a fundamen¬ tal number and X and Y elements resembling those of the Glossophaga group, but yet showed similarities with the Choeroniscus group in mor¬ phology of some autosomes.

Similar results are found in the studies of Phillips (1971) on denti¬ tions (adult and deciduous) in glos¬ sophagines. He recognized 1) a Glos¬ sophaga group of 10 genera, within which were included Glossophaga, Leptonycteris and Anoura, and 2) the Choeronycteris group including Choeroniscis.

The results of studies of gastric morphology and histochemistry pre¬ sented here indicate that there are two groups of glossophagines. The features of gastric morphology used as criteria for determining compo¬ sition of the two groups appears elsewhere in this discussion. The groups recognized are 1) Glosso¬ phaga, Anoura, and Leptonycteris and 2) Choeroniscus and Lichonyc- teris.

Although the groups presented here do not correspond precisely with those of Baker or Phillips, they most closely approximate those of the former author. Baker indi¬ cated that Anoura possessed chro¬ mosomes with features of the other two groups of glossophagines. The stomach of Anoura tends, in several respects, to be intermediate between those of the other two groups rec¬ ognized here. Examples of distin¬ guishing features include a larger and more elongate cardiac vestibule than in either Glossophaga or Lep¬ tonycteris, and a tendency toward intermediate staining of acid mu¬ copolysaccharide within the fundic and pyloric glands. As do Baker and Phillips, I consider the sub-

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Transactions Illinois Academy of Science

family Glossophaginae to be com¬ posed of at least two groups of spe¬ cies that probably represent an un¬ natural assemblage. Differences in gastric morphology are tentatively considered to be the result of early divergence in feeding habits. Stom¬ achs of the Glossophaga group more closely resemble those of obligate insect feeders than those of the Choeroniscus group. Stomachs of the latter assemblage possess many features found to occur otherwise only in fruit-eating species. This divergence probably involves un¬ known differences in quality or quantity of food ingested, or a com¬ bination of these factors. Thorough examination of feeding ecology and ethology should be undertaken to test this proposal.

Acknowledgements

I wish to thank Dr. Carleton J. Phillips for his helpful suggestions during final preparation of the manuscript. I also wish to acknowledge the help of Mr. Larry C. Watkins, who provided stomachs of mor- moopid bats. Some specimens were ob¬ tained under the sponsorship of a con¬ tract (DA-49-193-MD-221 5) from the Medical Research and Development Com¬ mand, U. S. Army, to Dr. J. Knox Jones, the University of Kansas. This study was supported, in part, by a grant from the Mary Ashby Cheek Research Fund, Rock- foxd College, to the author.

Literature Cited

Arata, A. A., J. B. Vaughn, and M. E.

Thomas. 1967. Food Habits of Certain

Colombian Bats. J. Mamm., 48(4) :653-

655.

Baker, R. J. 1967. Karyotypes of Bats of the Family Phyllostomidae and Their Taxonomic Implications. Southwestern Naturalist., 12(4) :407-428.

Barbour, R. W., and W. H. Davis. 1969. Bats of America. Univ. Kentucky Press.

286 pp.

Forman, G. L. 1971. Comparative His¬ tological and Histochemical Studies of Stomachs of Selected American Bats. Kansas Univ. Sci. Bull., in press. Goodwin, G. G., and A. M. Greenhall. 1961. A Review of the Bats of Trinidad and Tobago. Bull. Amer. Mus. Nat. Hist., 122 (Art. 3) :191-301. Hoffmeister, D. F., and W. W. Good- paster. 1954. The Mammals of the Huachuca Mountains, Southeastern Arizona. Illinois Biol. Monogr., 24:1-52. Lillie, R. D. 1965. Histopathological Technic and Practical Histochemistry. McGraw-Hill, New York, xii -j- 775 pp. Phillips, C. J. 1971. The Dentition of Glossopaginae Bats: Development, Morphological Characteristics, Varia¬ tion, Pathology, and Evolution. Univ. Kansas Publ., Mus. Nat. Hist., Miscel. Ser., No. 54, 136 pp.

Rouk, C. S., and B. P. Glass. 1970. Com¬ parative Gastric Histology of Five North and Central American Bats. J. Mamm., 51(3) :455-472.

Schultz, V. W. 1970. Eingie Bemerkun- gen zum Bau Verdauung straktes und der systematischen Stellung des Spitz- zahnflughundes, Harpionycteris white- headi Thomas, 1896 (Megachiroptera). Zeit. fur Saiigetierkunde, 35, Heft 2: 81-89.

Smith, J. D. 1971. Biosystematics of the Chiropteran Family Mormoopidae. Univ. Kansas Publ., Mus. Nat. Hist., in press.

Vaugh, T. A., and G. C. Bateman. 1970. Functional Morphology of the Forlimb of Mormoopid Bats. J. Mamm., 51(2): 217-235.

Manuscript received January 29, 1971.

STUDIES ON THE PHENOL-SOLUBLE LIPOPOLYSACCHARIDES FROM SERRATIA MARCESCENS BIZIO

I. ISOLATION AND CHEMICAL CHARACTERIZATION

JOSEPH C. TSANG

Department of Chemistry, Illinois State University, Normal, Illinois 61761

Abstract. — By means of ethanol pre¬ cipitation and Sephadex G-200 Gel filtra¬ tion, a lipopolysaccharide and a lipogly- coprotein were isolated from the phenol phase after the endotoxin complex of Serratia marcescens Bizio was submitted to 45% hot aqueous phenol extraction. The results of the chemical analyses of these two fractions indicated that the protein moiety may be covalently linked to the endotoxin molecule.

In the course of studying the iso¬ lation and fractionation of endo¬ toxin (lipopolysaccharide-protein complex) from Serratia marcescens Bizio (Tsang and Rilett, 1970), we submitted the purified endotoxin to hot aqueous phenol extraction (Westphal et al., 1952) with the purpose of removing the protein moiety from the endotoxin com¬ plex. However, our observation in¬ dicated that the effect of phenol treatment was more degradative than simple deproteinization. Most of the fatty acids and significant amounts of carbohydrates of the original endotoxin were removed along with the protein moiety in the phenol phase. Since several phenol-soluble lipopolysaccharides have been reported to occur in other Gram-negative bacteria (Raff and Wheat, 1968), it might be of interest to isolate and characterize the phenol-soluble materials from the endotoxin of S. marcescens Bi¬ zio. This report represents the study of isolation, fractionation, and chemical characterization of the phenol-soluble material. Electro¬ phoretic, immunochemical, and bio¬ logical studies of the various frac¬ tions will be reported subsequently.

Material and Methods

Serratia marcescens Bizio, grown on an inorganic medium, was sup¬ plied by General Biochemicals, Chagrin Falls, Ohio. The endotoxin was isolated and purified according to the procedures reported previ¬ ously (Tsang and Rilett, 1970). Phenol extraction of the endotoxin was performed according to the procedure of Westphal with 45% aqueous phenol at 65-70° C for 30 minutes (Westphal, et al., 1952). After the phenol phase was sepa¬ rated from the aqueous phase, it was washed three more times with equal volumes of water. The aque¬ ous phases were combined, dialysed exhaustively against water, and lyo- philized (Fr. LPS-A). Ethanol was added to the phenol phase in the ratio of 9.5:1 v/v (ethanol: phenol phase), and the mixture was al¬ lowed to stand overnight at 4° C. The precipitate (Fr. GP) was col¬ lected by centrifugation (3,000 rpm for 15 minutes) and washed con¬ secutively with acetone, acetone: ether (1:1 v/v), and finally ether.

Total neutral sugars, uronic acids, hexosamines, protein, and fatty acids were determined according to procedures reported previously (Tsang and Rilett, 1970; Tsang and Kallvy, 1971). Heptose was deter¬ mined by the method of Dische as modified by Osborn (Osborn, 1963). Phosphorus was determined by the method of Bartlett (Bartlett, 1959). The presence of 2-keto-3-deoxy- octonic acid was demonstrated qual¬ itatively by the thiobarbiturate

283

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method of Weissbach and Hurwitz (Weissbach and Hurwitz, 1959).

Paper chromatographic analysis of the hydrolyzed samples (2 N HC1 at 110° C for 4 hours) was performed by the descending meth¬ od on Whatman No. 1 filter paper with the following solvent systems:

A. Ethyl acetate :pyridine:wa- ter (3.6:1:1.5 v/v)

B . Butanol ipyridine :water (11:3.6:5.5 v/v)

Reducing sugars were detected by spraying with ammoniacal silver nitrate and p-anisidine hydrochlor¬ ide, while amino sugars and other amino compounds were detected by spraying with ninhydrin (in 0.2% butanol) .

Column chromatography of frac¬ tion GP was performed on Sepha- dex G-200 in 0.05 M pyridine-ace¬ tate buffer, pH 7.8. Fractions were collected every 3 ml. Aliquots of 0.2 ml were taken for protein and total carbohydrate analysis (an- throne method).

Results and Discussion

The phenol-soluble material (Fr. GP) isolated by ethanol precipita¬ tion from the phenol phase was a white amorphous solid. It was sol¬ uble in most of the aqueous buffers at slight alkaline pH, such as pyri¬ dine/acetate, pH 7.8, and borate buffer, pH 8.4. Paper chromato¬ graphic analysis after acid hydroly¬ sis revealed that there was no qual¬ itative difference in the sugar com¬ ponents between fraction GP and the corresponding lipopolysacchar- ide fraction LPS-A or the starting endotoxin fraction. Glucose, man¬ nose, galactose rhamnose, glucosa¬ mine, uronic acid, and heptose were identified. Heptose and 2-keto-3- deoxy-octonic acid were also de¬ tected by colorimetric methods. With the exception of uronic acids, the other sugars represented the sugar components of the basal core of bacterial lipopolysaccharide (Alaupovic, et al., 1966). Table 1 shows the chemical composition of

Table 1. — Chemical Composition of the Endotoxin Complex, the Lipopoly¬ saccharide (Fraction LPS-A), and the Phenol-Soluble Material (Fraction GP) of Serratia marcescens Bizio.

Endotoxin

Complex

%

Lipopolysaccharide (Fr. LPS-A)

%

Phenol-Soluble Material (Fr. GP)

%

Protein

8.0

10.0

30.5

Fatty Acids Anthrone-Positive

22.6

13.5

22.8

Carbohydrates

18.7

30.2

40.0

Glucosamine

2.4

0.9

3.6

Uronic Acids

6.9

11.2

10.6

Heptose

5.3

6.5

6.3

KDO

+

+

+

Phosphorus

1.19

1.35

2.25

the starting endotoxin, the phenol- treated lipopolysaccharide (Fr. LPS-A), and the phenol-soluble frac¬ tion (Fr. GP). The results of the chemical analyses clearly indicated that fraction GP is a complex mole¬ cule consisting of lipid, polysac¬ charide, and protein.

In order to study the homoge¬ neity of fraction GP and to demon¬

strate that this fraction was not a mixture of degraded components of lipids and carbohydrates or pro¬ teins, it was submitted to column fractionation on Sephadex G-200. Two distinct components were ob¬ tained (Figure 1). The major com¬ ponent was recovered in 90% yield from the exclusion volume. Rechro¬ matography of the major compo-

.20

Tsang — Phenol-Soluble Lipopoly saccharides

285

Fr. GP-1

\

- * - 1 - O - f- - ! - ( - 1 ■ I _

*+ 8 12 16 20 24 28 32

Tube Number

Figure 1. Sephadex G-200 Column Chromatography of Fraction GP. Vol¬ ume collected 3 ml per tube. Column (1.5 x 60 cm) was monitored by protein analysis (Lowry method). Similar result was obtained when column was moni¬ tored by carbohydrate analysis (anthrone method).

nent (Fr. GP-1) on Sepharose 4 B failed to provide further fractiona¬ tion.

The chemical composition of the subfractions are presented in Table 2. It appears that the fatty acids and carbohydrates removed in the phenol phase are eventually recov¬ ered in the form of complex mole¬ cule^). The analytical data show that the phenol-soluble material from the endotoxin of S. marcescens contained at least two components. The major component has the es¬ sential characteristics of bacterial lipopolysaccharide (Alaupovic, et al., 1966), while the minor com¬

ponent is possibly a lipoglycopro¬ tein (60% protein, 14% fatty acids, and 33% total carbohydrates) which is very similar to that of the phenol- soluble material isolated from the endotoxin of the wild-type chromo- genic S. marcescens 08 (70% pro¬ tein, 2% fatty acids, and 4% car¬ bohydrate) (Wober and Alaupovic, 1969). However, there are some chemical as well as physical chemi¬ cal differences between these two preparations (fraction GP-2 and lipoglycoprotein from 08 strain). The lipoglycoprotein from the wild- type (08) contained a pigment, pro- digiosin, and was insoluble in most acids and bases, while fraction GP-2 was readily soluble in most aqueous solvents. Recently, a prodigiosin containing cell envelope glycopro¬ tein was isolated from the cell walls of S. marcescens 08 (Tsang, et al., 1971; Tsang and Kallvy, 1971). It is not certain whether this glyco¬ protein is related functionally or structurally to the phenol-soluble lipoglycoproteins from the endo¬ toxin of either the wild-type (08) or the mutant (Bizio).

Since the starting endotoxin had been shown to be heterogeneous (Tsang and Rilett, 1970), it is diffi¬ cult to speculate at this stage the structural relationship of the iso¬ lated fractions (Fr. GP-1 and Fr. GP-2) to the endotoxin complex. However, the possible isolation of the lipoglycoprotein (Fr. GP-2) from the endotoxin complex may suggest

Table 2. — Chemical Composition of Sub-fractions.

Phenol-Soluble

Material (Fr.

GP) and Its

Fr. GP

Fr. GP-1

Fr. GP-2

%

%

%

Protein

30.5

26.5

60.0

Fatty Acids

22.8

20.6

14.1

Anthrone-Positive Carbohydrates

40.0

42.0

25.0

Glucosamine

3.6

3.3

1.8

Uronic Acids

10.6

11.8

6.0

Heptose

6.3

5.6

4.5

KDO

+

T

+

Phosphorus

2.25

1.50

1.80

286

Transactions Illinois Academy of Science

that the protein moiety of the en¬ dotoxin may be covalently linked to the lipopolysaccharide complex rather than associated through ionic or hydrophobic linkages. In other words, it is possible that the pro¬ tein moiety in bacterial endotoxin is specific to the endotoxin molecule and not of non-specific cellular ori¬ gin. It is hoped, however, that studies on the immunochemical cross-reactivity of the various frac¬ tions may reveal their possible func¬ tional and structural relationships to each other and to the endotoxin complex, as well as to the cell en¬ velope glycoproteins.

Acknowledgement

The author wishes to express his ap¬ preciation for the financial support for this investigation from Research Corpo¬ ration, Chicago, Illinois, and Illinois State University Research Committee.

Literature Cited

Alaupovic, P., A. C. Olson, and J. Tsang. 1966. Studies on the Charac¬ terization of Lipopolysaccharides from Two Strains of Serratia marcescens. Ann. N. Y. Acad. Sci. 133:546-565. Bartlett, G. R. 1959. Micromethod for Lipid Phosphorus Determination. J. Biol. Chem. 234:466-468.

Osborn, M. J. 1963. Studies on the Gram¬ negative Cell Wall. I. Evidence for the Role of 2-Keto-3-Deoxy-Octonate in the Lipopolysaccharide of Salmonella Typhimurium. Proc. Nat. Acad. Sci. (U.S.). 50:499.

Raff, R. A., and R. W. Wheat. 1968. Carbohydrate Composition of Phenol- Soluble Lipopolysaccharides of Citro- bacter freundii. J. Bacteriol. 95:2035- 2043. '

Tsang, J., and J. Rilett. 1970. Isolation and Fractionation of Lipopolysaccha¬ rides from Serratia marcescens Bizio. Trans. Ill. State Acad. Sci. 63:324-328.

_ _ S. Tattrie, and D. Kallvy.

1971. Outer Cell Envelope Glycopro¬ tein from Two Strains of Serratia mar¬ cescens. Applied Microbiology. 21:27- 31.

_ , and D. Kallvy. 1971. As¬ sociation of Prodigiosin with Outer Cell Wall Components. Trans. Ill. State Acad. Sci., 64: 22-26.

Weissbach, A., and J. Hurwitz. 1959. The Formation of 2-Keto-3-Deoxy- Heptonic Acid in Extracts of E. Coli B. J. Biol. Chem. 234:705-709.

Westphal, 0.,..0. Luderitz, and F. Bis¬ ter. 1952. Uber die Extraktion von Bakteriern mit Phenol/Wasser. Z. Na- turoforsch. 7B:148.

Wober, W., and P. Alaupovic. 1969. Studies on the Protein Moiety of En¬ dotoxins from Gram-negative Bacteria. Abstract of Papers, 158th American Chemical Society National Meeting, New York, No. 206.

Manuscript received December 16, 1970.

THE RESPONSE OF SOUTHERN ILLINOIS BARREN VEGETATION TO PRESCRIBED BURNING

ROGER C. ANDERSON AND JOHN SCHWEGMAN

Univ. of Wise. Arboretum, 1207 Seminole Highway, Madison, Wise. 53711 and III. Nature Preserves Commission, P. O.Box 661, Vienna, III. 62995.

Abstract. — Permanent quadrats were used to study the effect of prescribed burning on an area of barren vegetation that was being over-grown by honey¬ suckle ( Lonicera japonica), trees, and shrubs. The site was burned in the spring of 1969 and 1970. Herbaceous and woody plants were sampled on three occasions; in the fall before burning, and in August following each burn. Several species of prairie plants responded favorably to the burning. The prairie willow Salix humilus appeared to be spreading as a result of the burning. The only species to have tree size individuals (greater than 3.5 in. dbh) killed by the fire was Juniperus vir- giniana. However, seedlings of Juniperus virginiana and Betula nigra were elimi¬ nated. Following the second burn, but not the first, the frequency of honeysuckle was reduced by one-half. The second burn occurred later in the spring than the first after the honeysuckle had already be¬ gun growth.

Forests covered most of southern Illinois before settlement. Prairies were of limited occurrence and gen¬ erally did not extend much further south than Carbondale (Anderson, 1970; 1970a). “Barrens” occurred south of the Illinoian glacial limit and beyond the major area of prai¬ ries. Barrens apparently had more trees and shrubs than prairies and lacked some characteristic prairie species (Vestal, 1936) and might best be described as forest-prairie transitions. At presettlement times these areas were probably degraded forests that had been invaded by prairie plants as a result of fires.

A few of the broader ridges and stream valleys in the unglaciated portions of southern Illinois un¬ doubtedly supported these barrens as indicated by occasional stands of some prairie species. In the ex¬ treme southeastern portion of the state some of the areas recorded as

barrens in the original land survey now support closed forest commu¬ nities, a pattern repeated in many areas throughout the midwest as the result of the cessation of nearly annual fires set by aboriginals that had maintained these communities (Curtis, 1959; Vogl, 1964; Muir, 1965).

A vegetation map based on the surface soil color apparently pro¬ vides the best published informa¬ tion about these barren areas (Fehrenbacher and Alexander, 1958). The soils are transitional be¬ tween dark prairie and the lighter forest soils.

The purpose of this study was to carefully document changes in plant composition in response to fire using permanent quadrats. If the barren vegetation existed in presettlement times as the result of fire, then burning should tend to perpetuate this community. Fire might increase the dominance of prairie species; however, the woody plants should persist. The area studied was about 1/2 acre, on level ground adjacent to Burke Branch Creek in southern Pope County (S.W.1/4 sect. 4, T 15S, R6E). The area was badly en¬ croached by Lonicera japonica (Jap¬ anese honeysuckle) in addition to other woody species. A further ob¬ jective was to determine the effec¬ tiveness of fire as a management tool to control honeysuckle.

Methods

The area to be burned was sam¬ pled in November, 1968, and fired in the early spring March 16, 1969. A control plot was established and sampled with the burned area dur-

287

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Transactions Illinois Academy of Science

ing August, 1969. In 1970 the burn¬ ing was done later, April 5. Both the control and burned areas were resampled during the summer after the second burn, on the first of August. Throughout the study the control area showed little shift in composition or dominance of the plant species.

To record the response of woody plants to fire, the diameter of all tree species greater than 3.5 in. dbh were measured in four 1/40 acre and one 1/100 acre permanently marked circular quadrats. Seed¬ lings and saplings (tree species less than 3.5 in. dbh) as well as shrubs were sampled in five 1/100 acre circular quadrats that were nested within the tree quadrats. The seed¬ lings, saplings, and shrubs were placed into one of five size classes: (1) > 6 in. tall < 4.5 ft., (2) > 4.5 ft. tall < .5 in. dbh, (3) > .5 in. < 1.5 in. dbh, (4) > 1.5 in. < 2.5 in. dbh, (5) > 2.5 in. < 3.5 in. dbh.

Ten one meter square quadrats were nested within the 1/40 acre tree plots to sample herbs and woody plants less than six inches tall. In the 1/100 acre tree plot only four meter square quadrats were located. The honeysuckle was sampled by nesting five decimeter square quad¬ rats within each of the square meter quadrats.

Results and Discussion

The quadrat frequency (per cent

quadrats of occurrence) for the burned area in 1970, after two burns, is given in Table I for species hav¬ ing a frequency of 10 per cent or greater. Changes in frequency for selected species from 1968 to 1970 is shown in Figure 1 and discussed in detail later. The area is diverse with a total species list of 125 herbs and 26 trees and shrubs on about one-half acre. The flora is mixed, containing prairie species, wood¬ land and forest edge species as might be expected for barren vege¬ tation. The two bluestems, Andro¬ pogon gerardi and A. scoparius, were the dominant prairie grasses; how¬ ever, Sorghastrum nutans was prom¬ inent in scattered areas. Cassia fas- ciculata and C. nictitans were the dominant forbs.

The density of trees per acre on the permanent plots was 51.8 in 1968 and it remained unchanged throughout the study. The total tree basal area was 21.1 in 1971 and it increased slightly from 20.0 sq. ft. in 1968. Only four species of trees Ulmus alata, Juglans nigra, Platanus occidentalis, and Betula nigra had individuals greater than 3.5 in. dbh in the permanent tree quadrats. None of the trees of these species, ranging in diameter from 3.7 to 10.6 in. (dbh), were killed by the fire. However, tree sized red cedars, outside of the permanent plots, were eliminated by the burn¬ ing.

Table 1. — Frequency values for the burned area (1970) after two burns.

Andropogon gerardi

77.3

Cassia fasiculata

63.6

Andropogon scoparius

61.4

Cassia nictitans

56.8

Panicum anceps

47.7

Coreopsis tripteris

38.6

Acalypha gracilens

36.4

Solidago nemoralis

34.1

Ambrosia artemisiifolia

31.8

Potentilla simplex

29.5

Sorghastrum nutans

22.7

Euphorbia corollata

22.7

Rudbeckia hirta

20.5

Solidago altissima

18.2

Elymus virginicus

15.9

Cirsium altisimum

15.9

Stylosanthes biflora

13.6

Tradescantia ohiensis

11.4

Galium pilosum

11.4

Croton glandulosus

11.4

Panicum lanuginosum

11.4

Erigeron strigosus

11.4

Anemone virginiana

11.4

Uniola latifolia

11.4

Strophostyles leiosperma

11.4

Eupatorium serotinum

11.4

Anderson & Schwegman — Barren Vegetation

289

Burning decreased the number of woody stems less than 3.5 in. dbh in all but the smallest size class (6" tall but less than 4.5 feet tall) where there was a marked increase be¬ cause of resprouting, Figure 2. The total number of woody stems per acre in the smallest size class in¬ creased from 5,800 in 1969, to 9,100 in 1969, and in 1970 there were 10,160 stems per acre. However, the seedlings (size classes 1 and 2) of red cedar and river birch, were completely eliminated.

Three species were primarily re¬ sponsible for the increase of woody stems in the first size class, as a re¬ sult of resprouting: Salix humilis, Cornus ammonium, and Juglans ni¬ gra. Salix humilis, the prairie wil¬ low, increased the most of any of the species and showed a strong tendency to spread as the willow clumps became larger with repeated burning. The total number of wil¬ low stems, in the first size class, in¬ creased from 1,240 in 1968 to nearly 6,000 in 1970. The density of Rubus (probably R. ostryifolius ) stems greater than 6" tall remained about the same.

Herbaceous species that showed a marked increase in frequency after burning include several legumes, Cassia fasciculata, C. nictitans, Sty- losanthes bi flora, Strophostyles leio- sperma, two goldenrods Solidago nemoralis and S. altissima, and tall tick seed, Coreopsis tripteris, Figure 1. Ragweed (. Ambrosia artemisiifo- lia) increased after the first year of burning; however, it declined after the second burn in 1970. The ini¬ tial increase in ragweed and annual legumes may be in response to the more open environment following fire.

In another study still in progress (Van Valkenburg, 1971) the two species of Cassia showed a similar response and in one area increased from 300 individuals per acre to

33,000 with a single fire. Other le¬ gumes, including perennial ones, in¬ creased following the burn. Simi¬ larly, Wahlenberg et al. (1939) work¬ ing in Mississippi found that the number of legumes per acre was 41,500 on burned plots compared to 23,900 on unburned areas after nine years of annual burning. Lemon (1967) working in Africa also re¬ ported an increase in legume dens¬ ity in response to fire. The work being done in Southern Illinois by us and others (Johnson, 1969) indi¬ cates that there is an increase in the density of legumes following fire.

However, on the Tucker Prairie in Missouri, an increase in grass dominance with repeated burning was reported by Kucera and Koel- ling (1964) and Kucera (1970). They did not find an increase in legumes with burning.

Several of the species that show a marked reduction in frequency in response to fire included Panicum polyanthes, P. clandeslinum, Era- grostis spectabilis and Rubus sp. The Rubus is probably R. flagellaris that is less than six inches tall and was treated as a herb. Steyermark (1968) lists the two species of Panicum as occurring on moist ground and on wet prairies.

Figure 1 shows that Gaura biennis increased slightly in response to fire. These results do not show the ac¬ tual response to fire by this species. In several areas outside of the sam¬ pling plots Gaura increased much more than the data indicates. V e- ronicastrum virginicum also in¬ creased markedly outside of the area sampled.

Gentiana flavida (yellow prairie gentian) was restricted to one small clump that was being badly en¬ croached by honeysuckle at the ini¬ tial sampling in 1968. A permanent quadrat was located so that the gentian clone was included within it. The gentian responded well to

NO. STEMS / ACRE

290

Transactions Illinois Academy of Science

PER CENT FREQUENCY

Cassia fasiculata Cassia nictitans Coreopsis tripteris Solidago nemoralis Ambrosia artemisiifolia Solidago altissima Stylosanthes biflora Rudbeckia hirta Strophostyles leiosperma Gaura biennis Tradescantia ohiensis Anemone virginiana Panicum polyanthes Panicum ciandestinum Eragrostis spectabilis Rubus sp

10 30 50 10 30

—i - 1 1 1 1 - 1 - 1 - 1

1968

Figure 1. Changes in quadrat frequency for selected species in the study area.

the fire and the number of stems in the quadrat increased each year. In 1968 there were 5 gentian stems, in 1969, 10, and in 1970 there was more than a three-fold increase over 1968, 17 stems were in the quadrat.

Figure 2. The change in the total number of woody stems by size class.

An area that lacked any prairie vegetation grew up to a dense patch of fireweed ( Erechtites hieracifolia) . After the second burn this same area was dominated by Cirsium al- tissimum. However, the fireweed showed little tendency to invade areas that had prairie species well established.

A large portion of the viney honey¬ suckle that had climbed up into the trees was removed by the first burn. The fire burned up the vines on the trees to heights of 10-15 feet. While most of the honeysuckle in the trees was removed and did not re¬ establish itself on the trees, after the first fire, there was no decrease in quadrat frequency as much of the honeysuckle resprouted.

The first burn was in early spring, March 16, 1969. The following year the study area was burned on April 5, 1970, after the buds on the hon¬ eysuckle had just begun to burst. The frequency of honeysuckle was reduced by about 1/2 from 24 per cent to about 12 per cent, Figure 3. On the control area, established in

291

Anderson & Schwegman — Barren Vegetation

FREQUENCY OF LONICERA JAPONICA Based on

220 10cm X 10cm quadrats

24

1968 1969 1970

Figure 3. The response of Lonicera japonica to repeated fires.

1969, 50 decimeter quadrats were used to sample the honeysuckle. Its frequency remained almost the same, 68 per cent in 1969 compared to 66 per cent in 1970.

Prescribed burns appears to be an effective tool for controlling hon¬ eysuckle in locations where there is a vegetation such as prairie that will respond favorably to the burn¬ ing and offer the honeysuckle in¬ creased competition. A burn as late as possible in spring, preferably after the honeysuckle has begun growth but while the prairie plants are still dormant would seem to be the most effective.

Acknowledgements

The authors acknowledge the U. S. Forest Service for permitting the study to be done on the Shawnee National

Forest (Headquarters Harrisburg, Illi¬ nois) and for the loan of fire fighting equipment. Thanks are also due to Mrs. M. Rebecca Anderson for her assistance in data collection and Professor Grant Cottam (Univ. of Wis., Botany) for his review of the manuscript.

Literature Cited

Anderson, Roger C. 1970. Prairies in the prairie state. Trans. Ill. State Acad. Sci. 63 (2) :2 14-221.

_ , 1970a. Prairie restoration in

Southern Illinois. A paper presented at the Second Midwest Conference on prairies and prairie restoration, Madi¬ son, Wisconsin (Sept., 1970).

Curtis, J. T. 1959. The vegetation of Wisconsin. Univ. of Wise. Press, Madi¬ son, 657 pp.

Fehrenbacher, J. B. and J. D. Alexan¬ der. 1958. Native vegetation and sur¬ face soil colors in Illinois, Agronomy Facts, Univ. of Ill., College of Ag. Johnson, Richard. 1969. Personal Com¬ munications. Crab Orchard National Wildlife Refuge, Carterville, Illinois. Kucera, Clair. 1970. Ecological effect of fire on tall grass prairie. P. 12, Proceed¬ ing of a symposium on prairies and prairie restoration. Knox College Bio¬ logical Field Station, Pub. No. 3, Gales¬ burg, Ill.

Kucera, C. L. and M. Koelling. 1964. The influence of fire on the composi¬ tion of central Missouri prairie. Amer. Midland Nat. 72:142-147.

Lemon, P. C. 1967. Effects of fire on herbs of the southeastern United States and central Africa. Proc. Tall Timber Fire Ecol. Conf. 6:113-127.

Muir, J. 1965. The story of my boyhood and youth. Univ. of Wise. Press, Madi¬ son. 227 pp.

Steyermark, Julian. 1968. Flora of Mis¬ souri. The Iowa State Univ. Press, Ames, Iowa. 1728 pp.

Van Valkenburg, Charles. 1971. The response of Southern Illinois prairie type vegetation to burning. Ms. thesis in preparation.

Vestal, A. G. 1936. Barrens vegetation in Illinois, Trans. Ill. Acad. Sci. 29: 29-80.

Vogl, R. L. 1964. Vegetational history of Crex Meadows, a prairie savanna in northwestern Wisconsin. Amer. Mid¬ land Nat. 78:487-495.

Wahlenberg, W. G., S. W. Green and H. R. Reed. 1939. Effects of fire and cattle on longleaf pine lands as studied at McNeill, Mississippi. U.S. Dept. Agric. Tech. Bull., 683:1-52.

Manuscript received April 6, 1971.

ANALYSIS OF THE ELECTRON DENSITY AND POTENTIAL OF SOLID BENZENE

J. L. AMOROS AND MARISA CANUT-AMOROS

Materials Science Laboratory, Southern Illinois University, Carbondale, Illinois 62901

Abstract. — The electron density dis¬ tribution of the benzene molecule is ana¬ lyzed in terms of the selected electron shell (SES) method. With this method the electron density is split into two terms corresponding to the inner and outer elec¬ trons, respectively. The validity of sub¬ tracting a Gaussian distribution for the inner electrons is checked against the equivalence of subtracting the Is2 con¬ tribution. The potential function for the benzene crystal is calculated from experi¬ mental x-ray diffraction data. A low po¬ tential barrier is observed between the atoms in the benzene ring indicating the presence of delocalized electrons. Also the high potential barrier between molecules is in accordance with the idea that mole¬ cules are closed electron systems.

Benzene occupies a central posi¬ tion in organic chemistry and in molecular theory. The basic elec¬ tronic structure of the molecule is well understood in terms of the mo¬ lecular orbital theory, which in es¬ sence, considers the bonding elec¬ trons to be distributed into the a , or localized, and a , or delocalized, types. The molecule of benzene con¬ sists accordingly of a framework built up by the trigonal hybrid sp2 orbitals of the separated atoms over¬ lapping with themselves or with the Is orbitals of hydrogen. The linear combination of the pz atomic orbitals of the carbon atoms build up the a molecular orbitals. The electron density of benzene has been worked out theoretically by March (1952) by applying the Thomas- Fermi and the molecular-orbital method, but there is no other anal¬ ysis of the experimentally observed electron density except the work done by Cox et al. (1958). One of the problems in the study of the electron density via x-ray diffrac¬ tion methods is that it is difficult to get rid of the series termination

effect. Another problem is that the most important contribution to the total electron density derives from the electrons near the nucleus of the atom, forbiding any detailed analysis of the experimental maps.

In a series of papers, the authors have shown (Amoros and Canut- Amoros, 1968, 1969, 1970) that it is possible to separate the contribu¬ tion of the inner and outer electrons to the atomic scattering factor. As a consequence, the analysis of the density distribution from an experi¬ mental point of view can be done. The method, that has been called the selected electron shell (SES) method, is based on the possibility of an analytical representation of the atomic scattering factor as a Gaussian polynomial of positive terms, each one corresponding to what we call an electron shell. The method proved to be powerful in the sharpening of the Patterson of a molecular crystal, and it was tested successfully in the analysis of the electron distribution of the outer electrons in hexamine. It seemed, therefore, appropriate to extend the SES analysis to the molecule of solid benzene.

Crystal Structure of Solid Benzene

Benzene has a fairly simple or¬ thorhombic crystal structure, for which good x-ray diffraction data have been provided by Cox, Cruick- shank and Smith (1958) from ex¬ periments at -3°C. Solid benzene belongs to the space group Pbca of unit cell dimensions a = 7.46, b = 9.66, c = 7.03 A. It contains four molecules located on the centers of symmetry at 000, 0,1/ 2,1/2 ;l/2,0.

292

Amoros & Canut-Amoros — Solid Benzene

293

1/2, and 1/2, 1/2,0, as first given by Cox (1932). The refinement of the structure by the forementioned au¬ thors gave well resolved atomic po¬ sitions for the three independent carbon atoms. The positions of the hydrogen atoms were assumed to be at a radial distance of 1.084 A from the carbon atom to which the hydrogen is associated. The atomic coordinates of the hydrogen atoms were calculated in this way by Harada and Shimanouchi (1966).

An overall reliability factor of 10.5% was obtained by Cox and collaborators after five differential cycles of refinement, introducing anisotropic temperature factors. Those temperature factors were further utilized by the same au¬ thors to determine the translational and librational components of the amplitudes of vibration of the rigid benzene molecule. The possibility of this analysis showed us that the observed amplitudes and the whole crystal structure were reliable, and that the SES method could be ap¬ plied to this crystal. In our study we shall assume that the atomic coordinates as determined by Cox, Cruickshank and Smith are correct.

Application of the SES Method

In our previous work on hexa- mine (Amoros and Canut-Amoros, 1969) it was shown that the inner¬ most electrons of carbon could be approximated by a Gaussian func¬ tion whose Gs was taken as 2, the number of electrons in the inner shell. Also it was shown that the parameter g’s, which contains both the form and temperature factors of the Gaussian function, can be refined by using the structure fac¬ tors that correspond to reflections with J r* | > .9 A-1. The SES meth¬ od utilizes a difference Fourier sum¬ mation of the type

(r) = 1 X (F - (F ) Pouter ^ - h obs ' inner

V

exp (2/xi r.rp

(|r£|<R*lim) (1)

where v is the unit cell volume, Fobs are the observed structure am¬ plitudes, and Finner are the calcu¬ lated structure amplitudes for the inner electrons alone. Eq. (1) can be calculated by evaluating the contribution of the inner electrons once the parameter g’inner has been determined for each independent carbon atom. In the case of benzene 110 observations were used in the region of the reciprocal space be¬ yond | r* | = .9 A0"1. In the refine¬ ment of ginner the accepted atomic coordinates were used. The contri¬ bution of the hydrogen atoms in this region of reciprocal space is negligible and it was disregarded. Equation (1) is then calculated us¬ ing all terms within a region of the reciprocal space limited by a sphere of radius | r* | = l.A0-1, the limit of influence of the outer electrons of carbon. Series (1) contains there¬ fore the contribution of the outer electrons of carbon and the hy¬ drogen atoms and is free of series termination effect.

Once the Gaussian functions that describe the electron density of the inner electrons are known, the elec¬ tron density distribution of the in¬ ner electrons in the benzene mole¬ cule can be calculated either by convolution or by a Fourier sum¬ mation of the normal type whose coefficients are the calculated ones. Due to the high temperature factor of the carbon atoms in benzene, the Fourier series method is appli¬ cable. The series is practically con¬ vergent, and no important series termination effect is observed as

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Transactions Illinois Academy of Science

discussed later on. Finally, the dens¬ ity map of the outer electrons of the carbon atoms alone can be cal¬ culated by subtracting from Eq. (1) the contribution of the hydrogen atoms.

At this stage one may question the legitimacy of subtracting a Gaussian function as the corre¬ sponding function of the electron distribution of the innermost elec¬ trons in carbon. In benzene we have the carbon Is, 2s, and 2p, and the hydrogen Is functions. The ap¬

proximation used by the SES meth¬ od must be equivalent to subtract the Is state of carbon. In order to be sure that this is the case, the contribution of the Is function was subtracted using the values of the shell scattering of Is 1/2 given by Cromer and Waber (1964) taking into account that two electrons be¬ long to this state. The procedure followed in this new calculation was similar to the previous one. First the anisotropic temperature factors for the Is shell were deter-

Figure la. Electron density distribution of benzene. Section through the molecu¬ lar plane. Contour lines at increments of 0.2 e A-*. Linear scale 1 cm = 1A. Carbon inner electrons only. The first contour line 0.0 e A-» is not significative.

Amoros & Canut-Amoros — Solid Benzene

295

mined using the observed values for | r* | > .9 A0-1. Then a difference Fourier series of the type of Eq. (1) was performed in which the Firmer were computed in terms of the Is electrons alone.

Analysis of the Results

The most interesting maps are those corresponding to the section through the molecular plane. Ac¬ cording to our theory, the electron density can be split into two terms: the inner and outer electron density. Figure 1 gives the SES maps.

The first map (Fig. la) corre¬ sponds to the inner electrons. The zero line is very well defined, and no values higher than than 0.1 e. A°-3are observed in the map out¬ side the molecule. The fluctuation

Figure lb. Benzene outer electrons (a Gaussian core for carbon atoms has been subtracted). First contour line 0.2 e A-*.

of the background is so small that the map is practically free of series termination effect. The electron density shows maxima of about 3.65 e. A0-3 centered at the sites of the six carbon atoms of the molecule. Thermal motion causes an overlap of the electron density at mid-point between two adjacent carbon atoms,

and due to this a reading of 0.9 e. A°-3 is observed at that mid-point.

J

U

Figure lc. Benzene outer electrons (Is isotropic state for carbon atoms has been subtracted). First contour line 0.2 e A-s.

The outer electrons density maps are particularly interesting. Two kinds of maps have been computed. One map was calculated subtract¬ ing only the inner electrons as given by the Gaussian approximation (Fig. lb). The map contains then the electron distribution of the outer electrons of carbon and the hydro¬ gen electron. The map clearly shows the distortion of the basic hexagon of carbons produced by the pres¬ ence of hydrogens. It is interesting to note that the outer electrons of carbon behave like a doughnut of almost constant density of about 1.4 e. A0-3 showing very small max¬ ima of 1.5 e. A°-3. In order to check the validity of the Gaussian ap¬ proximation for the inner electrons, a similar map was computed by subtracting the contribution of the Is orbital. The resulting map (Fig. lc) is identical to the previous one (Fig. lb), the only difference being that the small maxima of 1.5 e. A-3 at the atomic positions are more de-

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fined. The size of the van der Waals envelope and distribution of con¬ tour lines is identical in both cases, as well as the count of 0.2 e. A-3 at the center of the benzene ring. From this follows that the Gaussian ap¬ proximation for the inner electrons is a realistic one. The maps are very similar to the ones calculated by March (1952), corresponding to the molecular orbital electron-density contours in a plane parallel to the planeo of the ring at a height of 0.35 A above the plane of the ring, which could be expected from the averaging effect of thermal vibra¬ tion in the real molecule.

/

O

Figure Id. Carbon outer electrons only. First contour line 0.2 e A-a.

The last map corresponds to the outer electrons of the carbon atoms alone (Fig. Id). In the calculations the contribution of the inner elec¬ trons of the carbon and the hy¬ drogen atoms have been subtracted. The consequences of existing de¬ localized electrons is the existence of a region of almost constant elec¬ tron density. In fact, the map shows very clearly that this is the case in benzene. It is important to note that the subtraction of the hydro¬ gen atoms did not alter the electron

density in the benzene ring nor the inner part, and only the humps due to the hydrogen atoms have disap¬ peared.

Better information can be ob¬ tained by observing the graph of Fig. 2. In the graph the electron density in the plane of the molecule has been plotted along a radial line uniting two opposite carbon atoms of the ring. The electron density curves of the inner, outer and hy¬ drogen electrons are clearly visible. Also, the positions of the C and H as deduced from x-rays by Cox, Cruikshank and Smith (1958) and neutron diffraction experiments by Bacon, Curry and Wilson (1964) are included. Thermal motion forces the observed peak positions of the carbon atoms as determined from x-ray experiments to move toward the center of the benzene ring. It is rewarding to note that the zero electrons contour line in our maps coincide with the effective size of the van der Waals envelope for car¬ bon atoms or for hydrogen + car¬ bon atoms determined from the in¬ term olecular potential (Newman, private communication).

The Potential Function

The study of the nature of the chemical bond can also be conveni¬ ently done by analyzing the poten¬ tial distribution in the unit cell. It was shown by Ewald (1938) and Laue (1940) that the electrostatic potential can be calculated from the Fourier series

(1NA hi exp (V r'rh4) (2)

h

where e is the electron charge. This series has the advantage that con¬ verges very rapidly, especially in molecular compounds where the temperature factor is normally high. Further advantage of Eq. (2) is that it can be calculated directly from

Amoros & Canut-Amoros — Solid Benzene

297

_3

3.65 e A

W-C IKTtRAaiON

Figure 2. Electron density along two diametrally opposite carbon atoms. Linear dimension 1 cm = 1A.

(i) Inner electrons of carbon atoms only.

(ii) Outer electrons with hydrogen atoms.

(iii) Outer electrons without hydrogen atoms.

(iv) Electron density of hydrogen atoms.

experimental data. Series (2) has been used for the analysis of the po¬ tential in semiconductors (see, for instance, papers published in Sirota (1968). However, no calculations have been done for molecular crys¬ tals, although this kind of function has great interest because it gives the experimental form of the van der Waals envelope of a molecule and enables the determination of the potential barrier between mole¬ cules. With this in mind, the ob¬ served potential function for ben¬ zene was computed.

The most significant map is the section through the molecular plane, which is given in Fig. 3. The atomic positions are determined by well defined potential wells separated by low potential barriers. The barrier has a potential of 1.6 eV. The low value of the potential barrier is in agreement with the view of exis¬ tence of delocalized electrons in the benzene ring. The center of the mo¬ lecule shows a barrier of 4.8 eV. with respect the atomic well, which

indicates the possibility of some electron density in the center of the molecule. This is in agreement with the SES map of the outer elec¬ trons that shows an electron dens¬ ity of 0.2 e. A-3 at that point.

One of the most interesting fea¬ tures of the map is the clear limita¬ tion of the benzene molecule, that is bound by a high potential bar¬ rier. The lowest value of this bar¬ rier corresponds to about 14 eV with respect to the atomic posi¬ tions. This value corresponds to the line along the nearest intermo- lecular distance in the plane of the section. This indicates that strong repulsion between the electron clouds of neighboring molecules exist and that the molecule must be considered as a closed system of electrons in agreement with cur¬ rent ideas of molecular crystals.

The value of the potential and potential barriers obtained from Eq. (2) depends upon the tempera¬ ture factor. Therefore, the actual values given here must be taken as

298 Transactions Illinois Academy of Science

Figure 3. Electronic potential distribution through the benzene molecular plane. The first continuous line corresponds to 0 e V. Contours at increments of 0.3 e V.

having qualitative rather than quantitative significance. However, a significative picture of the exis¬ tence of low and high potential bar¬ riers can be obtained from electron- potential maps.

The study of intermolecular po¬ tential barriers is of great interest for a better understanding of the chemistry of molecular solids. Fur¬ ther work on this sense is therefore necessary.

Computational Work The computational work was

made in an IBM 7044 and the plots in the digital incremental CalComp 565 plotter with off-line magnetic tape unit 470.

The least squares refinements and structure factor calculations were done with the ORFLS program of Busing, Martin and Levy (1962). The Fourier series were computed with the FORTRAN program of Zalkin (1962). Details of the actual computations involved in the SES method have been given by Canut- Amoros, Casper, Walters and Amoros (1970) and the computing

Amoros & Canut-Amoros — Solid Benzene

299

programs described in the Com¬ puting Report of Canut-Amoros et al. (1968).

Acknowledgements

We wish to thank, the Data Processing and Computing Center, Southern Illinois University for the facilities given to our research. Also we want to thank Mrs. Janice McIntyre for taking care of the preparation of the computations involved in this work.

This research was partially supported by the Solid State Sciences Division of the Air Force Office of Scientific Research, Grants AF-AFOSR-68-1587 and AF- AFOSR-67-0832B.

Literature Cited

Amoros, J. L. and Canut-Amoros, M. (1969) Analysis of density distribution of the outer electrons in hexamine. Z. Kristallogr. 129, pp. 124-141.

_ (1968) On the effect of the

electron shell structure of the atom in x-ray diffraction. Z. Kristallogr. 127, pp. 5-20.

_ (1970) The selected-electron-

shell method. Part I: Theory. Trans. Ill. State Acad. Science 63, pp. 117-124. Bacon, G. E., Curry, N. A. and Wilson, S. A. (1964) A crystallographic study of solid benzene by neutron diffraction. Proc. Roy. Soc. London, A 279, pp. 98-110.

Busing, W. R., Martin, K. O. and Levy, H. A. (1962) ORFLS, a FORTRAN crystallographic least-squares program. Oak Ridge Nat. Lab. ORNL-TM-305. Canut-Amoros, M., Casper, T. and Walters, C. (1968) A set of computing programs supporting the selected-elec¬ tron-shell method. Materials Science

Laboratory (S.I.U.) Report No. 1, pp. 1-143.

_ , and Amoros, J. L. (1970)

The selected-electron-shell method. Part II: Implementation. Trans. Ill. State Acad. Sci. 63, pp. 125-135.

Cox, E. G. (1932) Crystal structure of benzene. Proc. Roy. Soc. London, A 135:491-497.

_ , Cruickshank, D. W. J. and

Smith, J. A. S. (1958) The crystal struc¬ ture of benzene at -3°C. Proc. Roy. Soc. London, A 247:1-21.

Cromer, D. T. and Waber, J. T. (1964) Scattering factors computed from rela¬ tivistic Dirac-Slater wave functions. Los Alamos Scientific Laboratory, LA- 3056:1-222.

Ewald, P. P. (1938) Elektrostatische und optische Potentiale in Kristallraum und im Fourierraum. Ges. d. Wiss. Nachr. a. d. Phys., Astron., Geophys., Techn. 3:55-64.

Harada, I. and Shimanouchi, T. (1966) Normal vibrations and intermolecular forces of crystalline benzene and naph¬ thalene. J. them. Phys., 44:2016-2028.

Laue, M. von (1940) Zur Elektrostatik der Raumgitter. Z. Kristallogr. A 103: 54-70.

March, N. H. (1952) Theoretical deter¬ mination of the electron distribution in benzene by the Thomas-Fermi and molecular-orbital methods. Acta. Cryst. 5:187-193.

Sirota, N. N. (1968) Chemical bonds and thermodynamics in semiconductors. Consultants Bureau, New York. pp. 1-33.

Zalkin, A. (1962) A FORTRAN Fourier program. Lawrence Radiation Lab., California.

Manuscript received April 6, 1971.

PLANT INVESTIGATIONS II.

STUDIES ON THE HEXANE EXTRACT OF

Cl RSI UM ARVENSE

DAVID M. PIATAK AND LARRY S. EICHMEIER

Department of Chemistry, Northern Illinois University, DeKalb, Illinois 60115

Abstract. — The hexane extract of Cir- sium arvense was investigated by applica¬ tion of column, thin layer, and gas chrom¬ atography and spectroscopic methods. A C18-C31 alkane mixture, B-amyrin, tarax- asterol, taraxasterol acetate, tetracosanol, hexacosanol, and octacosanol were posi¬ tively identified.

Cirsium arvense, or Canadian this¬ tle, the curse of the Midwestern farmer, is a plant which has re¬ ceived little attention by the natu¬ ral product chemist (Shelyuta et ah, 1970). Throughout the chemical lit¬ erature the multitude of articles about it has centered upon its eradi¬ cation although there have been scattered reports on its potential as a supply of blood coagulants (Maz- zanti, 1954), seed growth inhibitors (Helgeson and Konzak, 1950), and meat-curing solution additives (Jen¬ sen and Hess, 1951). The little phy¬ tochemical work done with the C7r- sium genus has been concerned only with the isolation of flavanoids (Wagner et al., 1960; Nakaoki and Morita, 1959; Ibid., 1960; Morita and Shimizu, 1963) and the detec¬ tion of alkaloids, tannins, and fla¬ vanoids (Ismailov, 1958; Bandyu- kova and Shinkarenko, 1965; Ko- lodziejshi, et al., 1966).

In this paper we describe our pre¬ liminary investigations on the al¬ kane, n-alcohol, and terpene com¬ ponents of Cirsium arvense.

Experimental

General — Melting points (mp) were taken on a Fisher-Johns apparatus and are corrected. Infrared spectra were recorded with a Beckmann IR-8, nuclear magnetic resonance spectra (n.m.r.), with a Varian A-60 on CDCI3 solutions, and mass spec¬

tra with a Perkin-Elmer-Hitachi RMU-6E instrument. Silica gel HF254 (Merck) at a thickness of 0.5 mm was used for all thin layer chromatography (TLC). A Varian Aerograph Series 200 instrument with a flame ionization detector was used for gas liquid chromatography (GLC). The instrument was utilized at an injector temperature of 300°, a detector temperature of 325°, and a nitrogen rate of 80% flow at 65 psi. A 5 ft. by 1/8 in. SS column packed with 5% SE-30 on acid- washed dmcs 60/80 chromosorb W was used for all separations. Preliminary Separation — The aerial portions of C. arvense were collected during June 1968 in DeKalb. The air-dried material (2 kg) was pow¬ dered and then exhaustively ex¬ tracted with hexane in a Soxhlet apparatus. Removal of the hexane gave a residue (89 g) which was treated in 400 ml. of tetrahydro- furan with 120 ml. of 10% KOH. After heating on the steam bath for 15 minutes the resulting solution was diluted with water, and the or¬ ganic materials were recovered with ether. The residue (63 g.) was then dissolved in hexane and the hexane removed in vacuo. This process was repeated twice more to ensure full removal of all other solvents.

The treated extract was dissolved in hexane and chromatographed on 900 g of Alcoa F-20 alumina which had been neutralized to Activity II-III by stirring it under hexane for 3 days with 27 ml. of 10% acetic acid. The column was eluted with 1.5 liter fractions. The solvent used and the fractions are as fol-

300

Piatak & Eichmeier — Hexane Extract Cirsium

301

lows: hexane 1-8, 2% benzene-hex¬ ane 9-11, 5% benzene-hexane 12-14, 10% benzene-hexane 15-17, 25% benzene-hexane 18-48, 35% ben¬ zene-hexane 49-59, 50% benzene- hexane 60-68, benzene 69-75, 1% chloroform-benzene 76-80. The chromatogram had been continued with other solvents but no identi¬ fiable material was isolated. Taraxasterol Acetate — A white solid which was shown by TLC analysis to be essentially one compound was found in Fractions 5-9. The ma¬ terial was chromatographed pre- paratively on three TLC plates with benzene. Recrystallization of the re¬ covered material (400 mg.) from methylene chloride gave colorless crystals, mp 220-225°; [a]" 96.7° (C,0.2); max(in KBr) 1735,’ 1642, 898 cm-1; n.m.r. 278 (d,2H,J = 1.0), 122(s,3H), 63, 61, 58, 57, 51 c.p.s.; m/e 468(M+), 453, 408, 399, 393, 357, 339, 297, 249, 218, 205, 204, 203, 191, 190, 189 (base).

Anal. Calcd. for C32H5202: C, 81.99; H, 11.18. Found: C, 81.70; H, 11.12.

Reported mp and [a]D values for taraxasterol acetate have been 234- 248° and 95.0° (Kasprzyk and Py- rek, 1968); 238-240° and 96.0° (Chaudbury and Ghosh, 1970); 256- 257° and 100° (Simonsen and Ross, 1957).

n- Alcohols — Fractions 20 (410 mg) and 21 (430 mg) were purified by treatment with Norit A. Crystal¬ lization of each from benzene-meth¬ anol yielded material melting at 75-77.5° and 80-82°, respectively. GLC analysis of both at a column temperature of 253° gave peaks cor¬ responding to 1-tetracosanol at 146 sec., 1-hexacosanol at 238 sec., and 1-octacosanol at 394 sec. Both frac¬ tions consisted mainly of 1-octa¬ cosanol and were almost identical in alcohol proportions. Identifica¬ tion of latter two alcohols wa^ made by comparing their GLC behavior

to a commercial sample of 1-hexa¬ cosanol and the 1-octacosanol sam¬ ple obtained in the first paper of this series (Piatak and Reimann, 1970). The 1-tetracosanol structure was inferred from the retention time pattern.

Taraxasterol — By TLC analysis fractions 23-48(7. Og.) were found to contain one compound. Recrys¬ tallization of the material from ben¬ zene, acetone, and chloroform-meth¬ anol after Norit A treatment gave

I. 1 g. of colorless crystals, mp 203- 205° (normal) and 213-215° (evacu¬ ated capillary); [a\22D 90.9° (C,3.0); v max (in KBr) 3400, 1642, 885 cm-1; n.m.r. 277 (d,2H, J = 2.00), 64, 61, 58, 56, 51, 46 c.p.s.; m/c 426(M+), 411, 408, 357, 315, 218, 207, 189 (base).

Anal. Calcd. for C30HO50: C, 84.44; 11.81. Found: C, 84.37; H,

II. 76.

Literature values for the melting point and specific rotation of ta¬ raxasterol were 213-219° and 82.0° (Kasprzyk and Pyrek, 1968); 222- 225° and 93.5° (Atherinos et al., 1962).

The taraxasterol was converted to an acetate which was identical to the taraxasterol acetate obtained above and to a benzoate, mp 230- 232 [reported mp 242-244° (Simp¬ son, 1944)].

fi-Amyrin — TLC analysis was used to determine that fractions 49-59 (1.5 g.), 60-68(1.2 g.), and 69-80 (0.9 g.) contained taraxasterol and another major component. The two were separated by preparative TLC using 25% ethyl acetate-hexane. GLC of the new compound at a column temperature of 235° re¬ vealed that the material consisted of two closely related compounds with retention times of 954 sec. and 1096 sec. The latter compound was identified as j3 -amyrin by compari¬ son of its retention time with au¬ thentic material and by noting an

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Transactions Illinois Academy of Science

increase in the amount of response for the second peak when authentic material was added to the mixture. The more mobile material was not investigated further when compari¬ son of its GLC characteristics to several triterpenes failed to identify it.

n -Alkanes — Fractions 1-4 (24g.) were rechromatographed on 720 g of Woelm neutral alumina of Ac¬ tivity I. The first three fractions eluted (3 x 500 ml. hexane) were combined to give 500 mg. of semi¬ crystalline material, max no C = O absorption. GLC analysis with a programmed column temperature of 90-280° at a rate of 4-6°/min in¬ dicated the presence of 18 separate substances.

The mixture was dissolved in benzene and treated with a drop of bromine. Reanalysis of the material after removal of the solvent showed 15 substances. By comparing the remaining peaks to a set of stan¬ dards identified by mass spectral analysis of gas chromatographically separated material (Reimann, 1970) and extrapolation of the regularity in the retention times, the alkanes were identified. The mixture was found to consist of the following n- alkanes: Ci8(4.4%), Ci9(<l), C20 (<1), C2i(<l), C22( < 1) , C23(5.8),

c24(1.0), C25(4.1), C26(1.9), C27(25.3),

C28(2.8), C29(37.8), C3o(1.7), C3i (12.4).

Discussion

Our work with Cirsium arvense appears to be the first report con¬ cerned with the identification of in¬ dividual compounds in this species. Although Shelyuta et al. (1970) have explored this plant phyto- chemically, they had only concerned themselves with the classes of com¬ pounds present or not present.

Thus far, we have identified sev¬ eral known compounds. The first compound isolated was taraxasterol acetate, a derivative of a penta-

cyclic triterpene commonly found throughout the plant world. In fact, taraxasterol seems to be the major pentacyclic triterpene in C. arvense , since it was detected in a number of the chromatography fractions as the free alcohol or its acetate.

Taraxasterol itself appeared much further along in the column chromatography fractions. It was not readily identified at first even though it gave a single peak in GLC analysis, because the mp and spe¬ cific rotation data were widely di¬ vergent from literature values. How¬ ever, spectroscopic data did point to taraxasterol or a similar triter¬ pene since an exocyclic methylene group was indicated by n.m.r. and infrared spectra, a hydroxyl group by an infrared spectrum, and a pentacyclic skeleton by a mass spec¬ trum. Some derivatives were pre¬ pared, but the physical constants could not be fully reconciled with literature values.

The structure was finally resolved by a comparison of taraxasterol acetate with samples provided by other workers in India and Poland. Although a mixture m.p. with these samples had the same value as our material, the mass spectra and GLC characteristics of each were in full agreement, thereby fully establish¬ ing the structure.

The taraxasterol acetate isolated from the column appears to be an artifact produced by the action of acetic acid used to neutralize the column alumina on taraxasterol. Normally, acetates of triterpenes are not found readily in any quan¬ tity in plants. Since we did hydro- lize the plant extract with base be¬ fore chromatography, it is even less likely the acetate was originally present.

The isolation and identification of the mixture of straight chain primary alcohols obtained from the column after taraxasterol acetate

Piatak & Eichmeier— Hexane Extract Cirsium

303

was easily accomplished by various chromatographic techniques. Al¬ though the fractions were mixtures of practically the same composi¬ tion, the difference in their mp’s and the sharpness of the mp’s were not totally unexpected. In the first paper of this series (Piatak and Reimann, 1970) it had been found that mp’s of alcohols were not satis¬ factory criteria for identification and that GLC comparison to ma¬ terial fully established by a mass spectrum was best.

A second pentacyclic triterpene, P-amyrin, was also uncovered in this plant. This terpene, however, could not be isolated owing to the presence of another compound. Al¬ though several different methods, e.g. silver nitrate impregnated TLC plates and double-dip TLC tech¬ niques, were attempted, a clear cut separation between the two could not be gotten. The -amyrin never¬ theless was verified by the standard GLC technique of adding authen¬ tic material to the chromatographic sample and observing an increase in the response for that material.

The last series of compounds to be investigated were the n-alkanes. Even though these compounds were contained in the first four column fractions, they were mixed with several other materials. A second column chromatography employing a larger alumina to material ratio did give a colorless, crystalline mix¬ ture of alkanes containing a few unsaturated compounds. The un¬ saturated hydrocarbons could be removed readily by bromine treat¬ ment. Since the bp of the bromo addition product would be quite high, the net result of the bromine treatment is a removal of all un¬ saturated compounds. Of course, the peaks obtained for each alkane must be examined carefully to see if any have been increased by the addition products; but, thus far,

this has not been our experience. Acknowledgements

We wish to thank the National Science Foundation (GB7424) and the Seiffert Trust administered by the Illinois Divi- sion-American Cancer Society for partial support of this work. The assistance of K. Nusbaum and L. Neas, N.S.F. Student Science Training Program participants, in the initial stages of this project is greatly appreciated. We wish to thank Dr. S. K. Talapatra, Calcutta University, and Dr. J. Kasprzyk, University of War¬ saw, for supplying samples of taraxas- terol acetate.

Literature Cited

Atherinos, A. E., I. E. El-Kholy, and G. Soliman. 1962. Chemical Investiga¬ tion of Cynara scolymus L. Part I. The steroids of the Receptacles and Flowers. J. Chem. Soc.:1700-1704.

Bandyukova, V. A., and A. L. Shin- karenko, 1965. Paper chromatogra¬ phic determination of flavanoids in high altitude plants of the Teberda Reserva¬ tion. Farmatsevt Zh., 20:37-41. Chaudhury, N. A., and D. Ghosh. 1970. Taraxasterol and other triterpenoids in Capparis sepiaria leaves. Phytochemis¬ try, 9, 1885.

Helgeson, E. A., and R. Konzak. 1950. Phytotoxic effects of aqueous extracts of field bindweed and of Canada thistle. N. Dakota Agr. Expt. Sta. Bull., 12: 71-76.

Ismailov, N. M. 1958. Alkaloid contain¬ ing plants from the Khaldan and Ag- dash regions. (Boz Dagrange) Azer- baidzhan S.S.R. Izvest. Ahad. Nauk Azerbaidzhan S.S.R., Ser. Biol, i SeT skokhoz:ll-16.

Jensen, L. B. and W. R. Hess. 1951. Meat-curing pickle solutions contain¬ ing antibiotics. United States patent 2,550,253.

Kasprzyk, Z. and J. Pyrek. 1968. Tri- terpenic alcohols of Calendula officinalis L. flowers. Phytochemistry, 7:1631- 1639.

Kolodziejski, J. A. Mruk-Luczkiewicz, and D. Pietrzok 1966. Composition of Cirsium oleraceum. Farm. Polska, 22: 87-90.

Mazzanti, L. 1954. Antihemorrhagic ac¬ tion of Cirsium arvense. Boll. soc. ital. biol. sper., 30:267-269.

Morita, N., and M. Shimizu. 1963. Me¬ dicinal Resources. XXI. Flavanoids of Cirsium Plants in Japan. 3. Compo¬ nents of the leaves of Cirsium marti- mum. Yakugaku Zasshi, 83:615-618.

304

Transactions Illinois Academy of Science

Nakoaki, T., and N. Morita, 1959. Me¬ dicinal Resources XIII. Flavanoids of Cirsium. 1. Components of the leaves ofC. microspecatum, C. Otaijae, C. Yoslii- zawae, C. Japonicum, and C. purpura- tum. Yakugaku Zasshi, 79:1338-1340.

_ 1960. Medicinal Resources.

XIV. Flavanoids of Cirsium. 2. Compo¬ nents of the leaves of C. inundatum and C. kagamontanum. Yakugaku Zasshi, 80:1296-1299.

Piatak, D. M., and K. A. Reimann. 1970. The Isolation of 1-Octacosanol from Euphorbia corollata. Phytochem¬ istry, 9:2585-2586.

Reimann, K. A. 1970. Phytochemical Study of Euphorbia corollata. M.S. the¬ sis, Northern Illinois University.

Shelyuta, V. L., Y. I. Kolisnichenko, and N. T. Bubon. 1970. Phytochemi¬ cal Investigation of Cirsium arvense. Farm. Zh. (Kiev), 25:60-64.

Simonsen, J., and W. C. J. Ross. 1957. The Terpenes. Vol. IV. Cambridge Uni¬ versity Press, Cambridge, ix + 524 pp.

Simpson, J. C. E. 1944. The Triterpene Group. Part XI. The non-saponifable matter of Lactucarium germanicum, J. Chem. Soc.:283-286.

Wagner, H., L. Horhammer, and W. Kirchner. Flavones of Compositae and Papilionactae. I. Occurrence of pec- tolinarin and linarin in the plant king¬ dom. Arch. Pharm., 293:1053-1062.

Manuscript received March 2, 1971.

Notes

305

LONGEVITY RECORD FOR PIPISTRELLUS SUBFLAVUS

HARLAN D. WALLEY & WILLIAM L. JARVIS

Department of Biology, Northern Illinois University, DeKalb, Illinois 60115

Abstract. — Longevity record for Pipistrellus subflavus extended from 11.2 years to 14.8 years, with remarks on tooth wear.

The longevity of American bats has recently been reviewed by Paridiso & Greenhall (1967) from data compiled by the U. S. Fish & Wildlife Service band¬ ing returns. Fourteen species, represent¬ ing six genera are cited. The oldest pres¬ ent known longevity record is for a speci¬ men of Myotis lucifugus recovered 24 years after banding (Griffin & Hitchcock, 1965), while Hall, et al. (1957) recorded 18.5 years for Myotis keenii. Paridiso & Greenhall (1967) gave equally long dates for Eptesicus fuscus, 19 years and 16.5 years for Plecotus townsendii.

On 14 February 1971 and again on 25 February 1971, while recording recover¬ ies of Pipistrellus subflavus in South Black¬ ball Mine, 1.75 miles west of Utica, La¬ Salle Co., Illinois, a male P. subflavus was found carrying the U. S. Fish & Wildlife Service Band Number 57-04984, which was banded on 16 February 1957 at South Blackball Mine, by Dr. Wayne H. Davis. This would give this bat a mini¬ mum age of 14.8 years, since the young of these bats are born in June or early July. Paridiso & Greenhall (1967) gave 11.2 years as the greatest age for P. subflavus, while Davis (1966) using multiple regres¬ sion analysis showed that P. subflavus could live up to 13 years, but gave no indication of having recovered a speci¬ men of this age.

It is interesting to note that all of the species having exceedingly long life spans (M. lucifugus, M. keenii, M. sodalis, Ple¬ cotus townsendii, and Eptesicus fuscus in

eastern United States) bear only one young. Pipistrellus subflavus which is smaller than any of the above noted spe¬ cies has two young, and possibly this could be a major factor in its being shorter-lived.

In examining the canine teeth of this specimen on 25 February 1971, I was surprised to find only slight wear in this individual. This would agree with Twente (1955) for five year old bats. Hall, et al. (1957) have shown that tooth wear in M. lucifugus and M. keenii, as proposed by Twente (1955) and Stegeman (1956) for other species, is a highly unreliable cri¬ terion for age determination. The tooth wear criterion is apparently unreliable for P. subflavus also.

Literature Cited

Davis, W. H. 1966. Population dynamics of the bat Pipistrellus subflavus. J. Mammal., 47(3) :383-396.

Griffin, D. R. and H. B. Hitchcock. 1965. Probable 24-year longevity rec¬ ords for Myotis lucifugus. J. Mammal., 46:332.

Hall, J. S., R. J. Cloutier and D. R. Griffin. 1957. Longevity records and notes on tooth wear of bats. J. Mam¬ mal., 38 (3) :407-409.

Paradiso, J. L. and A. M. Greenhall. 1967. Longevity records for American bats. Amer. Midi. Nat., 78 (l):251-252. Stegeman, L. C. 1956. Tooth develop¬ ment and wear in Myotis. J. Mammal., 37 (1) :58-63.

Twente, J. W., Jr. 1955. Aspects of a population study of cavern-dwelling bats. J. Mammal., 36 (4):379-390.

Manuscript received March 1, 1971.

306

Transactions Illinois Academy of Science

NOTES ON ILLINOIS AND WISCONSIN RESUPINATE

BASIDIOMYCETES

ANTHONY E. LIBERTA

Illinois State University, Normal

Abstract. — -Four species of resupinate Basidiomycetes are reported from either Illinois or Wisconsin for the first time.

Collections of fungi chat extend their known North American range are noted here. These collections were made either by me (AEL) or by R. M. Miller (RMM) and are now deposited in the mycological herbarium at Illinois State University (ISU).

Cerinomyces pallidus Martin

On decayed deciduous wood, Funks Grove, McLean County, Ill., VI. 20. 1969, AEL 1470 (ISU 1210); near Crab Orchard Lake, Williamson County, Ill., XI. 23. 1963, AEL 557 (ISU 688).

The reported North American range of this species now includes, in addition to Illinois, Iowa and Ontario (Martin, 1952) as well as Wisconsin and Colorado (Li- berta, 1965, 1966).

Galzinia incrustans (Hoehn. & Litsch.) Farm.

On decayed deciduous wood, Funks Grove, McLean County, Ill., XII. 1.1970, RMM 70-21 (ISU 1260).

The only other midwestern state where collections of this species have been re¬ ported is Missouri, under the binomial Corticium roseopallens Burt (1926).

Hyphodontia alutacea (Fr.) Eriks.

On decayed wood ( Tsuga ?), near Say- ner, Vilas County, Wisconsin, VII. 16. 1964. AEL 493 (ISU 1209).

This is the first report of the species in Wisconsin. Larsen (1964) has reported it from several eastern and western states

as well as from Canada. Miller and Boyle (1943) reported its occurrence in Iowa.

Xenasma grisellum (Bourd.) Liberta

On decayed deciduous wood, near Crab Orchard Lake, Williamson County, Illi¬ nois, XI.23. 1963, AEL 1454 (ISU 1211).

This species is apparently wide spread in Europe, but the only previous North American report was from the province of Quebec, Canada (Liberta, 1966a). The collection of this species in Illinois con¬ siderably extends its known North Ameri¬ can range.

Literature Cited

Burt, E. A. 1926. The Thelephoraceae of North America. XV. Corticium. Ann. Missouri Bot. Gard. 13:173-354. Larsen, M. J. 1964 Hyphodontia alu- taceae in North America. Can. Jour. Bot. 42:1167-1172.

Liberta, A. E. 1965. Notes on Wiscon¬ sin resupinate Basidiomycetes. My- cologia 57:459-464.

_ 1966. Notes on Colorado re¬ supinate Basidiomycetes. Trans. Ill. State Acad. Sci. 59:392-393.

_ 1966a. Resupinate Hyme-

nomycetes from Gaspe and adjacent counties (Canada) I. Mycologia 58: 927-933.

Martin, G. W. 1952. Revision of the North Central Tremellales. Univ. Iowa Stud. Nat. Hist. 19(3):1-122.

Miller, L. W. and J. S. Boyle. 1943. The Hydnaceae of Iowa. Univ. Iowa Stud. Nat. Hist. 18(2):l-92.

Manuscript received April 8, 1971.

Notes

307

MORRIS M. LEIGHTON 1887-1971

Dr. Morris M. Leighton, widely-known geological scientist and administrator, died Thursday, January 7, 1971, at Ur- bana, Illinois, at the age of 83, after a long illness. He was Chief of the Illinois State Geological Survey from 1923 until his retirement in 1954, during which time the Survey came to a preeminent position among state surveys.

Dr. Leighton was born in 1887 at Well¬ man, Iowa, a son of Stephen T. and Jane Leighton. He was educated at the Uni¬ versity of Iowa, where he received B.A. and M.S. degrees in 1912 and 1913, and at the University of Chicago, where he received his doctorate in 1916.

Between 1915 and 1923, Dr. Leighton was on the geology faculty at the Univer¬ sity of Washington, Iowa State Teachers College, Ohio State University, and the University of Illinois. During the period 1919-1923 he was also on the staff of the Illinois State Geological Survey. After his retirement he remained active profession¬ ally until shortly before his death.

Dr. Leighton published numerous scien¬ tific articles over the past 53 years and is recognized especially for his publications on Pleistocene glacial deposits. Particu¬ larly noteworthy was his contribution on the concept of a state geological survey as a research institution. The Illinois State Geological Survey was the first state survey to emphasize a coordinated research effort to bring together the vari¬ ous disciplines, e.g., geology, chemistry, physics, and engineering, and apply them to natural resources for the shaping of sound policies of conservation.

Dr. Leighton was a member of many scientific and technical societies including the Society of Economic Geologists of which he was past president and honor¬ ary member; the Geological Society of America, fellow and councilor; the Ameri¬ can Association of State Geologists, hon¬

orary member and past president; the American Geological Institute, director and past president; Illinois State Museum Board, member and chairman; Illinois Postwar Planning Commission, member and vice chairman; Advisory committee, U. S. Geological Survey; Coordinating Committee on National Water Policy; Chicago Geographic Society, fellow; American Association for the Advance¬ ment of Science, past vice president; Illi¬ nois State Academy of Science, past presi¬ dent; and Illinois Mining Institute, past president.

Dr. Leighton was also made an honor¬ ary member of the American Association of Petroleum Geologists, the Chicago Academy of Sciences, and the Illinois Mining Institute. He also served as busi¬ ness editor for “Economic Geology” for many years, and as editor of the State Geologists Journal.

Among his awards and honors, he was named Distinguished Alumnus at the University of Iowa in 1947, was elected a fellow of the American Academy of Arts and Sciences in 1952, was awarded an Honorary Doctor of Science Degree from Southern Illinois University in 1954, and was a member of the United States Dele¬ gation to the Twentieth International Geological Congress held in Mexico City in 1956.

He was a member of the Wesley United Methodist Church and of the Chaos, Dial, and University Clubs.

Dr. Leighton was preceded in death by rive brothers and one sister. He leaves his widow, Ada B. Leighton; a sister, Mrs. Golda Jenkinson of Tulsa, Oklahoma; three sons, F. Beach of Hacienda Heights, California; Morris W. of Seaforth, New South Wales, Australia; and Richard T. of Rockford; and 10 grandchildren. — Boris Musulin, Department of Chemistry, Southern Illinois University, Carbondale, Illinois.

Manuscript received March 1, 1971.

308

Transactions Illinois Academy of Science

GILBERT H. CADY 1882-1970

Gilbert Haven Cady, world-renowned coal scientist, died Friday, December 25, 1970, at Champaign, Illinois, at the age of 88, following a short illness.

Dr. Cady served as Senior Geologist and Head of Coal Section of the Illinois State Geological Survey from 1926 until his retirement in 1951 but remained ac¬ tive professionally until two weeks before his death. He was born in 1882 in Chicago. He studied at the Lewis Institute, Chi¬ cago, and Northwestern and Yale Uni¬ versities and the University of Illinois, and received his doctorate from the Uni¬ versity of Chicago in 1917.

He worked with the Illinois State Geo¬ logical Survey from 1907 to 1919, leaving to spend a year as a geological consultant in the interior of China. From 1920 to 1926, he was head of the geology depart¬ ment at the University of Arkansas.

His many scientific papers, published over a period of nearly 60 years, concen¬ trated on the field of coal geology. A high percentage of coal scientists in North America from time to time had profes¬ sional association with Dr. Cady.

Dr. Cady was a member of many other scientific and technical societies, includ¬ ing the Society of Economic Geologists, of which he was past president and coun¬ cilor; Geological Society of America, fel¬ low and counsilor, and American Associa¬ tion for the Advancement of Science, fel¬ low. He was a member of Phi Beta Kappa and Sigma Xi.

Dr. Cady was honored with a life mem¬ bership in the Illinois Mining Institute.

In 1963, the International Committee for Coal Petrology recognized his many years of contributions to this field with the pre¬ sentation of the Reinhardt Thiessen Medal for Coal Petrology. He was the third recipient of the award, established in 1953, and the only American so hon¬ ored to date.

North American coal petrographers gathered in Urbana in 1964 for the pre¬ sentation of a formal testimonial to Dr. Cady, and in 1967 he was the recipient of the Penrose Medal of the Society of Economic Geologists for “unusual origi¬ nal work in the earth sciences.”

Colleagues throughout North America are planning to establish a memorial fund to be used for recognition of contributions in coal geology.

Dr. Cady was a member of the Wesley United Methodist Church and the Ur¬ bana Exchange Club. He was preceded in death by his wife, Marian, and two sons, Gilbert and Allan. He leaves two daugh¬ ters, Ruth Adams of Urbana and Mary Johnson of Las Vegas, Nevada, and two grandsons, Derek and Cady Johnson, of Las Vegas. — Boris Musulin, Department of Chemistry , Southern Illinois University, Carbondale, Illinois.

Manuscript received March 1, 1971. Erratum:

In the article “The first record in Illi¬ nois of a population of Stethaulax mar- moratus (Say) (Hemiptera: Scutelleridae) with information on life history. 64:198- 200, the photograph for Figure 1 is re¬ versed, the male is to the left.

New Yo

rk Botanical Garc

en Library

3 51!

15 0034'

45*

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PREPARATION OF MANUSCRIPTS FOR THE TRANSACTIONS

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Doe, J. H. 1951. The life cycle of a land snail. Conchol., 26(3): 21-32, 2 tables, 3 figs.

Doe, J. H., and S. H. Jones. 1951. Mineralogy of Lower Tertiary deposits. McGraw-Hill Book Co., New York, iv -f- 396 pp.

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Department of Biological Sciences, Northern Illinois University,

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I

Transactions

of the

Illinois

State Academy

of Science library

APR 7 1972

new YORK

botanical garden

Volume 64 No. 4 1971

TISAAH

TRANSACTIONS of the ILLINOIS STATE ACADEMY OF SCIENCE

Editorial Board :

Malcolm T. Jollie, Northern Illinois University, Editor and Chairman

Stanley H. Frost, Northern Illinois University, Geology Gordon C. Kresheck, Northern Illinois University, Chemistry John C. Shaffer, Northern Illinois University, Physics Darrell L. Lynch, Northern Illinois University, Microbiology Robert H. Mohlenbrock, Southern Illinois University, Botany

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Mailing date, March _ , 1972

TRANSACTIONS

OF THE

ILLINOIS STATE ACADEMY OF SCIENCE

VOLUME 64 - 1971

No. 4

Illinois State Academy of Science

AFFILIATED WITH THE

Illinois State Museum Division Springfield, Illinois

PRINTED BY AUTHORITY OF THE STATE OF ILLINOIS

(36150-1650—3-72)

Richard B. Ogilvie, Governor

December 1, 1971

CONTENTS

Effect of Ethylene and 2, 3, 5-Triiodobenzoic Acid on Soybean Seedlings Grown in Hydroponic Culture

by E. Patrick Lira and Arthur H. Freytag .

Effects of Soil Moisture Stress on Foliar Nutrients of Loblolly Pine

by A. R. Gilmore .

Contributions to an Illinois Flora No. 4. Compositae II. (Tribe Heliantheae, Part I - Dyssodia, Helenium, Gaillardia, Hymenoxys, Hymenopappus, and Polymnia)

by R. P. Wunderlin .

Contributions to an Illinois Flora No. 5. Compositae III.

(Tribe Heliantheae, Part II - The Genus Coreopsis)

by R. P. Wunderlin .

Aryl Acetoacetates - Infrared and Ultraviolet Spectra by Forrest J. Frank, John H. Long and Terry K. Reid .

Methods of Evaluating Thickness and Texture of Glacial Till in Studies of Ground-Water Recharge

by Kemal Piskin .

The Pileate Pore Fungi of Illinois

by Robert W. Schanzle .

A Census of Mould Spores in the Atmosphere

by Hansel M. DeBartolo, Jr .

Inhibition of Leukocyte Induced Germination and Toxin Release From Clostridium botulinum Type A Spores by Chlorocresol by J. B. Suzuki, A. Benedik, and N. Grecz .

Ecological Study of a Hillside Marsh in East-Central Illinois

by Hampton M. Parker and John E. Ebinger .

The Effect of Dimethyl Sulfoxide on Human Erythrocyte Membrane

by Dennis M. Kallvy and Joseph C. Tsang .

Lake Shore Erosion

by Wyndham J. Roberts .

Local Effects of the New ACS Curriculum

by Shelba Jean Choate Musulin and Boris Musulin .

The Stabilization of a Gully by Natural Forest Succession

by Robert A. Bullington .

309

313

317

328

334

337

347

350

358

362

370

374

383

388

NOTES

A New Locality and a New Host Record for Trichodina discoidea Davis,

1947 in Southern Illinois

by Lawrence J. Blecka . 398

A New Distribution Record for Lycopodium flabelliforme in Illinois

by Loy R. Phillippe . 399

An Albino De Kay’s Snake ( Storeria dekayi Wrightorum Trapido)

From Central Illinois

by James H. Thrall . 400

Harlow Burgess Mills 1906 - 1971

by Robert A. Evers . 401

EFFECT OF ETHYLENE AND 2,3,5-TRIIODOBENZOIC ACID ON SOYBEAN SEEDLINGS GROWN IN HYDROPONIC CULTURE

E. PATRICK LIRA AND ARTHUR H. FREYTAG

Growth Sciences Center, International Minerals and Chemical Corp.

Libertyville, Illinois 60018

Abstract — Soybean seedling, Glycine max (L) Wayne, grown in hydroponic cul¬ ture were treated with ethylene and 2,3,5- triiodobenzoic acid by root application. Significant aerial shoot elongation with minimal lateral bud initiation and growth resulted from the ethylene treatment. The concomitant addition of 2,3,5-triiodoben- zoic acid to the ethylene treatment had an overriding effect on these growth re¬ sponses.

Treatment of root systems of plants with growth regulators has been the subject of several reports (Scott and Norris, 1970; Sherbeck,

1967) . Ethylene treatment of ex¬ cised root sections, root tips and intact seedlings has been reported to cause effects which resemble aux¬ in response (Chadwick and Burg, 1967, 1970; Radin and Loomis, 1969). Recent observations indicate this may not be generally true (Scott and Norris, 1970; Andreae, et al.,

1968) . The exogeneous ethylene treatment of root systems of intact plants without concomitant treat¬ ment of the aerial portions of the plant has been reported recently (Smith and Russell, 1969). We can now describe a previously unre¬ ported ethylene-induced response of soybean seedlings, Glycine max (L) var. Wayne, grown in hydroponic culture to which ethylene was added and removed without contamina¬ tion of the surrounding atmosphere. Under these conditions significant aerial shoot elongation with mini¬ mal lateral bud initiation and growth were observed. We have also overridden this shoot elonga¬ tion by the addition of 2,3,5-tri- iodobenzoic acid (TIBA) to the nu¬ trient solution.

Materials and Methods

Soybean seeds planted in vermic- ulite were germinated in the green¬ house and grown to the unifoliate leaf stage. Two healthy seedlings were then transplanted to a 3.7 liter widemouth glass jar covered by a five-holed, grooved, opaque plastic lid. The jar contained 3.6 liters of half strength Hoagland solution (Hoagland and Arnon, 1938), continually aerated with car¬ bon-filtered air. The container was covered completely with an opaque plastic film. The seedlings were sup¬ ported by sponge rubber gaskets. Two sintered glass tubes were in¬ serted in the lid in rubber stoppers, one for aeration and the other for ethylene, and an exhaust tube was extended from the remaining open¬ ing.

Each test consisted of four treat¬ ments replicated five times. The treatments were: control, ethylene, TIBA, and ethylene plus TIBA. Although four tests varying the TIBA concentration (1, 2.5, 5 and 10 ppm) were run, only the data of the tests at 1 and 5 ppm are re¬ ported, because all except the low¬ est concentration gave similar re¬ sults. The treatments were initi¬ ated when the seedlings were trans¬ planted and were terminated at the seven-eight trifoliate leaf stage. The nutrient solution and the TIBA were replaced weekly. Ethylene was added daily for a two-hour period at the same time each day. Dis¬ tilled water was added daily to maintain liquid level.

Research grade ethylene from a

309

310

Transactions Illinois Academy of Science

high pressure cylinder was distrib¬ uted equally to the jars by a mani¬ fold and flow meter at a rate of four liters/minute. The exhaust gas was collected by a manifold and vented to the exterior of the green¬ house. During the addition of ethy¬ lene this process was assisted by a low volume vacuum pump.

The ethylene concentration in the aqueous solution was determined by removing 2 ml solution with a syringe fitted with a six inch No. 20 needle. Samples were taken at 0.5, 2.0, 4.0, 7.0 and 24.0 h after initiation of ethylene addition. The samples were transferred to septum- capped 250 ml erlenmeyer flasks containing 1 g lithium chloride and were allowed to equilibrate for 24 hours. The vapors were analyzed

by gas chromatography (Burg and Burg, 1966).

Results and Discussion

The aqueous ethylene concentra¬ tion was calculated by application of the simple gas laws. The concen¬ tration varied with time (Figure 1). The solution was saturated (Mc- Auliffe, 1966) during the second h of addition. Then there was a rapid decrease to 10 ppm after 4 h fol¬ lowed by a more gradual reduction to 1 ppm after 7 h. Ethylene was still detectable (0.01 to 0.001 ppm) after 24 hr. The observed elonga¬ tion of the main shoot of soybean seedlings when ethylene treatment is rigorously confined to the root zone has not been previously de¬ scribed (Holm, et al, 1970). It can

Time (hours)

Figure 1. Variation of aqueous ethylene concentration with time. The variation between samples is illustrated by the vertical bars. The results are the composite of thirty samples at each sampling period. This cycle is repeated each day.

Lira and Frey tag — Growth Regulators in Soybeans

311

Table 1. Growth of Soybean Seedlings Treated with Ethylene and Tiba

Grown in Hydroponic Culture

Lateral

Lateral

Green

Weight

Treatment*

Height*

Bud No.

Length

Roots

Aerials

Control*

Test I

Ethylene

TIBA (5 ppm) Ethylene -j- TIBA (5 ppm)

100

+ 25a -21b -5b

100

-290b — la -15a

100

-300c -f- 80a + 3b

100

-53b

-26b

-32c

100

— 53bc -22b -170c

Test II

Ethylene

TIBA (1 ppm) Ethylene -j- TIBA (1 ppm)

-}-32a — 9c + 8b

-900c + 3a -21b

-160c

T 6a — 9ab

-24b — 17ab -62c

-25b

-41b

-91c

*Each datum is the mean of 10 plants. Two plants per treatment (jar) replicated five times. (See text for complete description of procedure.) The datum is expressed as a percent of control. Numbers for each test within a column followed by a different letter are significantly different at the 5% level using Duncan’s multiple range test.

be seen (Table 1) that the treated plants are between 25 and 32% taller than the controls. The two other unreported tests terminated at the three-four and thirteen tri¬ foliate leaf stages showed height increases of 52 and 12%, respec¬ tively. It appears that this response is most pronounced during the ear¬ lier stages of growth while at later stages the controls may actually be growing faster. A further observa¬ tion is the dramatic reduction in the number of lateral buds being formed and their subsequent growth. These results may be due to an increase in the production and distribution of gibberellins in the roots (Jones and Phillips, 1967) or perhaps to a redistribution of auxin (Beyer and Morgan, 1970; Abeles, 1966).

The response of soybean seed¬ lings to TIBA in hydroponic cul¬ ture has been described (Ohki, 1968) and our results (Table I) generally agree. When TIBA, an auxin trans¬ port inhibitor (Huffman, et at., 1967) is added at 5 ppm to the ethylene solution, elongation is completely overridden and lateral bud forma¬

tion is enhanced. However, TIBA at 1 ppm only partially reverses these ethylene-induced effects. These TIBA-ethylene interaction observations tend to indicate that root-applied ethylene is influencing the auxin transport system rather than a specific gibberellin system.

The overall growth in all the treatments was less than the con¬ trol. In general, the combined treat¬ ment caused a greater reduction in growth. The ethylene- treated roots gave a typical auxin growth re¬ sponse (Scott and Norris, 1970). Finally, in one of our tests (unre¬ ported) the seedlings became chlo¬ rotic, but the ethylene treated plants appeared significantly greener.

Literature Cited

Abeles, F. B. 1966. Effect of Ethylene on Auxin Transport. Plant Physiol., 41:946-948.

Andreae, W. A., Venis, M. A., Jursic, F., and Dumas, T. 1968. Does Ethylene Mediate Root Growth Inhibition by Indole-3-Acetic Acid? Plant Physiol., 43:1375-1379.

Beyer, E. M., and Morgan, P. W. 1970. Effect of Ethylene on the Uptake, Dis¬ tribution, and Metabolism of Indole- acetic Acid-l-14C and -2-14C and Naph-

312

Transactions Illinois Academy of Science

thaleneacetic Acid-l-14C. Plant Phy¬ siol., 46:157-162.

Burg, S. P., and Burg, E. A. 1966. The Interaction Between Auxin and Ethyl¬ ene and its Role in Plant Growth. Proc. U.S. Nat. Acad. Sci., 55:262-269.

Chadwick, A. V., and Burg, S. P. 1967. An Explanation of the Inhibition of Root Growth Caused by Indole-3-Ace- tic Acid. Plant Physiol., 42:415-420.

_ 1970. Regulation of Root

Growth by Auxin-Ethylene Interac¬ tion. Plant Physiol., 45:192-200.

Hoagland, D. R., and Arnon, D. E. 1938. The Water-Culture Method for Growing Plants without Soil. Calif. Agr. Exp. Sta., Circ. 347:1-39.

Holm, R. E., O’Brien, T. J., Key, J. L., and Cherry, J. H. 1970. The influence of Auxin and Ethylene on Chromatin- directed Ribonucleic Acid Synthesis in Soybean Hypocotyl. Plant Physiol., 45:41-45.

Huffman, C. W., Godar, E. M., and Torgeson, D. C. 1967. Inhibition of Plant Growth by Halogenated Benzoic Acids. J. Agr. Food Chem., 15:976-979.

Jones, R. L., and Phillips, I. D. J. 1967. Effect of CCC on the Gibberellin Con¬ tent of Excised Sunflower Organs. Plan- ta, 72:53-59.

McAuliffe, C., 1966. Solubility in Water of Paraffin, Cycloparaffin, Olefin, Ace¬ tylene, Cycloolefin and Aromatic Hy¬ drocarbons. J. Phys. Chem., 70:1267- 1275.

Ohki, K. 1968. Effects of Root Absorbed 2,3,5-Triiodobenzoic Acid on Nutrient Absorption and Growth of Soybean. Plant Physiol., Plant Physiology Meet¬ ing Suppl. 43:5-48.

Radin, J. W., and Loomis, R. S. 1969. Ethylene and Carbon Dioxide in the Growth and Development of Cultured Radish Roots. Plant Physiol., 44:1584- 1589.

Scott, P. C. and Norris, L. A. 1970. Separation of Effects of Auxin and Eth¬ ylene in Pea Roots. Nature, 227:1366- 1367.

Sherbeck, T. G. 1967. The influence of 2,3,5-Triiodobenzoic Acid (TIBA) in Fertilizer Bands on the Growth and Development of Soybeans ( Gylcine max.). Unpublished Doctoral Disserta¬ tion, University of Purdue, pp. 134.

Smith, K. A., and Russell, R. S. 1969. Occurrence of Ethylene, and its Sig¬ nificance, in Anaerobic Soil. Nature, 222:769-771.

Manuscript received April 19, 1971.

EFFECTS OF SOIL MOISTURE STRESS ON FOLIAR NUTRIENTS OF LOBLOLLY PINE

A. R. GILMORE

University of Illinois, Champaign-U rbana

Abstract — Loblolly pine ( Pinus taeda L.) seedlings were subjected to three soil moisture stresses during their second growing season. Foliar nitrogen was high¬ er and foliar potassium, calcium, and magnesium were lower in those seedlings grown in the drier treatment. Foliar phos¬ phorus was not affected by soil moisture stress.

Leaf analysis as a means of eval¬ uating the nutritive status of forest trees has met with varying success. Even where a correlation has been found between nutrient concentra¬ tions in the leaf and in the soil, there is always the question of how accurately such data reflect the nutrient supplying power of a given site. Many variables, in addition to soil fertility, affect the concentra¬ tion of chemical elements in the leaf, such as physiological maturity, position in tree, sampling date, time of day sampled, and others (Leyton and Armson, 1955).

These reported studies and con¬ ceptions indicate that soil-moisture stress must play an important role in the nutrient concentrations in tree foliage. In an attempt to clarify these findings and concepts, loblolly pine seedlings were grown during their second year in the greenhouse under varying soil-moisture stresses and the elements in the needles correlated with soil moisture.

Methods

The study consisted of two pha¬ ses: The first phase was conducted with one-year-old loblolly pine seed¬ lings planted in pots filled with a silt loam soil. This soil (Ap horizon) was developed from loess and is medium to high in K (.02 percent) ; low in P (trace), N (.01 percent), and organic matter (0.90 percent),

and moderately to strongly acid (pH 5.2). The equivalent of 1,000 kilograms per hectare of a 12-12-12 (N, P, K) fertilizer was applied to each pot at planting time. The second phase of the study conducted the following year was with year- old seedlings grown from seed in pots (three seed per pot) filled with the same soil as used in the first phase but not fertilized. During both years, the soil with seedlings was allowed to dry until about 30 percent, 50 percent, or 70 percent of the available soil moisture was exhausted as determined by weigh¬ ing, and then watered to above field capacity. Available moisture in this study is that moisture held in the soil between 1/3 and 15 bars as determined by the pressure pot and pressure membrane apparatus. There was 23 percent moisture in the soil at 1/3 bar and 8 percent moisture in the soil at 15 bars for this soil. Hereafter, these three soil- moisture levels will be known as low, medium, and high soil-mois¬ ture stresses, respectively.

Thirty seedlings were planted in individual pots in each of the three treatments in the first phase. Mor¬ tality reduced the number in the medium and high soil-moisture stress treatments to 18 and 8 seed¬ lings, respectively. Twenty -five pots were used in each treatment in the second phase, but the number of seedlings per pot varied from one to three. At the termination of the second experiment, there were 44, 47, and 40 seedlings in the low, me¬ dium, and high soil-moisture stress treatments, respectively.

At the end of both growing sea-

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Transactions Illinois Academy of Science

sons, plants were removed from the pots and dried at 75° C. Foliar ni¬ trogen was determined by a semi¬ micro Kjeldahl procedure (Ranker, 1927), phosphorus by phosphomo- lybdic acid (A.O.A.C., 1945), po¬ tassium by flame emission, and cal¬ cium and magnesium by the EDTA method (Malmstadt and Hadjiio- annou, 1959).

Analysis of variance was made of the five elements in the needles with soil-moisture stresses.

Results

Shoot growth of seedlings dur¬ ing the study period varied by mois¬ ture treatments. In both phases of the study, seedlings subject to the drier conditions grew about 90 per¬ cent (as a percent of total height at beginning of season), whereas those in the moist treatment (low soil moisture stress) grew about 120 percent.

Table 1 shows the average nu-

Table 1. Foliar Nutrient Concentrations of Seedlings Grown under Different

Moisture Stresses in the First Phase

Variable

Moisture stress

Low

Medium

High

Percent of O.D. Weight

N1

2.566

2.823

2.759

P

0.135

0.128

0.110

K1

0.280

0.241

0.243

xLow is significantly different from other stresses at 5 percent level.

trient levels in the foliage under high fertility in the first phase. When the soil was supplied with adequate moisture (low moisture stress), the nitrogen content of the needles was significantly lower than in seedlings growing at higher soil- moisture stresses. At this relatively high soil fertility, soil moisture did not influence the concentration of phosphorus in the needles, but plants growing at low moisture stress had more potassium in the

needles than plants growing under drier conditions.

Nutrient concentration in the needles are shown in Table 2 for the second phase. The results for N, P, and K were very similar to those obtained in the first phase, except that the concentrations of N and P were much lower and K higher. Calcium and magnesium concentrations were higher at the low moisture stress than in the

Table 2. Average Foliar Concentrations and Ratios of Concentrations

in Second Phase of Study

Variable

Moisture stress

F ratio1

Low

Medium

High

Percent of O.D. weight

N

0.862

1.093

1.118

16.32

P

0.072

0.078

0.071

1.85

K

0.319

0.301

0.290

3.45

Ca

0.286

0.266

0.264

4.07

Mg

0.231

0.208

0.204

12.40

Significant differences between stresses at 1 percent level, 2.99.

Gilmore — Effects on Foliar Nutrients

315

higher stresses in this phase of the study.

Discussion and Conclusions

The increase in foliar nitrogen in the high moisture stress treat¬ ment probably means that when growth was limited by soil mois¬ ture, nitrogen tends to accumulate within the plant because the rate of entry is approximately maintained in conjunction with the decreased rate of utilization in growth pro¬ cesses. The general tendency for potassium, calcium, and magnesium to be relatively low in plants on drier soil (high moisture stress) in¬ dicates that the rates of entry into the plant of these elements are less than the rates of utilization in the slow growing plants.

The increase in foliar potassium in this study with decreasing soil moisture stress agrees with results obtained for herbaceous plants and for 26-year-old red pine growing on a coarse sandy soil in New York State (Jurgensen and Leaf, 1965). But the small difference in foliar potassium for the two years is diffi¬ cult to explain, as it was thought that the potassium contents of seed¬ lings grown in the well-fertilized soil would be greater than they were.

It is well known that fertilizers interact with soil moisture stresses (Schomaker, 1969) and that the nu¬ trient concentration of plants grown at different fertility levels may not always conform to a set pattern. These results tend to confirm the thought that many factors influ¬ ence an element in a plant, and it is difficult to predict how any one factor can affect the concentration of an element within a plant.

It is recognized that the method used in reporting the concentration of nutrients in this paper (as a per¬ centage of weight of oven-dry fo¬ liage) has its limitation, and there is a possibility that some other

method such as total uptake might be superior. This possibility has been suggested for red pine in west¬ ern Massachusetts (Hoyle and Mader, 1964) and in the Adiron¬ dack region of New York (White and Leaf, 1965).

The most useful role that foliar analysis can play at the present time is in the nutrition of forest trees on depleted soils where nu¬ trient deficiencies have been indi¬ cated. This technique has been used quite successfully in New York on potassium-deficient soils (Madg- wick, 1964) and in Florida on phos¬ phorus-deficient soils (Pritchett and Llewellyn, 1966).

Acknowledgements

A portion of this research was supported by funds from the Illi¬ nois Agricultural Experiment Sta¬ tion, Hatch Project 55-341.

Literature Cited

Association of Official Agricultural Chem¬ ists. 1945. Official methods of analysis. Sixth ed., pp. 127-128.

Hoyle, M. C., and D. L. Mader. 1964. Relationship of foliar nutrients to growth of red pine in western Massa¬ chusetts. For. Sci. 10:337-347.

Jurgensen, M. F., and A. L. Leaf. 1965. Soil-moisture-fertility interactions re¬ lated to growth and nutrient uptake of red pine. Soil Sci. Soc. Amer. Proc. 29:294-299.

Leyton, L., and K. A. Armson. 1955. Mineral composition of the foliage in relation to the growth of Scots pine. For. Sci. 1:210-218.

Madgwick, H. A. I. 1964. The chemical composition of foliage as an index of nutritional status of red pine ( Pinus resinosa Ait.). Plant and Soil 21:70-80.

Malmstadt, H. V., and T. P. Hadjiioan- NOU. 1959. Rapid and accurate auto¬ matic titration method for determina¬ tion of calcium and magnesium in plant material with EDTA titrant. J. Agr. Food Chem. 7:418-420.

Pritchett, W. L., and W. R. Llewel¬ lyn. 1966. Response of slash pine to phosphorus in sandy soils. Soil Sci. Soc. Amer. Proc. 30:509-519.

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Transactions Illinois Academy of Science

Ranker, E. R. 1927. A modification of the salicylic-thiosulfate method suit¬ able for the determination of total ni¬ trogen in plants, plant solutions, and soil extracts. J. Assoc. Off. Agr. Chem. 10:230-251.

Schomaker, C. E. 1969. Growth and foliar nutrients of white pine seedlings as influenced by simultaneous changes

in moisture and nutrient supply. Soil Sci. Soc. Amer. Proc. 33:614-618.

White, D. P., and A. L. Leaf. 1965. Soil and tree potassium contents related to tree growth: II. Tissue K as deter¬ mined by total tree analysis techniques. Soil Sci. 99:109-114.

Manuscript received April 19, 1971.

CONTRIBUTIONS TO AN ILLINOIS FLORA No. 4. COMPOSITAE II. (TRIBE HELIANTHEAE, PART I DYSSODIA, HELENIUM, GAILLARDIA, HYMENOXYS, HYMENOPAPPUS, AND POLYMNIA).

R. P. WUNDERLIN

Department of Biology , St. Louis University, St. Louis, Mo. 63101+

Abstract. — A key to the 23 Illinois genera of the tribe Heliantheae and keys, descriptions, and distributional maps for the Illinois species of Dyssodia, Helenium, Gaillardia, Hymenoxys, Hymenopappus, and Polymnia are given.

This is the second of a series on the Compositae in Illinois. The first, a treatment of the tribe Vernonieae, has been published previously (Wunderlin, 1968).

This study includes only a key to the Illinois genera of the tribe Heliantheae and a treatment of the first six genera, Dyssodia, Helenium, Gaillardia, Hymenoxys, Hymeno¬ pappus, and Polymnia, in Illinois. Treatments of the other genera in the tribe will follow in subsequent publications. The magnitude of this tribe as well as the desirability of making the works available with¬ out too much delay has made it de¬ sirable to publish it in several parts.

The tribe Heliantheae is charac¬ terized by having opposite (rarely alternate or whorled) leaves, pre¬ dominately radiate heads, the style- branches usually hispidulous and with the stigmatic lines poorly de¬ fined, and the anther bases obtuse to sagittate (rarely caudate).

The tribe Helenieae has been tra¬ ditionally distinguished from the Heliantheae by a single technical character, the absence of chaff on the receptacle. This character is not always clear-cut; e.g., Gaillardia has a bristly rather than a naked re¬ ceptacle. The tribe Helenieae is now considered by most Compositae spe¬ cialists as an artificial taxonomic group. The sub tribes of the Helen¬ ieae apparently have been derived independently from the Heliantheae

in several different lines. Therefore, the Helenieae should not be con¬ sidered as a separate tribe but should be included in the Heliantheae as has been done in this treatment. The subtribe Ambrosiinae has be¬ come adapted to wind pollination and differs from the rest of the Com¬ positae in its much reduced corol¬ las and free or nearly free anthers. This group might justifiably be considered as a separate tribe or even a separate family as it has been treated by several other au¬ thors. However, the Melampodinae furnishes a good series of transi¬ tional forms between the typical Heliantheae and the less modified genera, e.g., Iva, of the Ambrosi¬ inae. Thus the Ambrosiinae are treated as part of the Heliantheae as is more customary.

The Heliantheae in Illinois is composed of 23 genera of which three, Gaillardia, Cosmos, and Galin- soga, have been introduced. Gail¬ lardia is from the west, Cosmos is from the southern United States, and Galinsoga is from the tropics. Several species of other genera have been introduced or are adventive in Illinois.

The distributional maps are based upon specimens housed in the fol¬ lowing herbaria: Eastern Illinois University, the Field Museum of Natural History, the Illinois Nat¬ ural History Survey, the Illinois State Museum, the Missouri Bo¬ tanical Garden, Southern Illinois University, the University of Illi¬ nois, and Western Illinois Univer¬ sity.

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The author is grateful to the cu¬ rators of the herbaria used in this study for allowing him to study their specimens. The author also gratefully acknowledges Dr. James R. Wells, Cranbrook Institute of Science, Bloomfield Hills, Michi¬ gan, for his assistance and com¬

ments on the treatment of Poly- mnia, Dr. Warren P. Stoutamire, the University of Akron, Akron, Ohio, for his comments on Gaillar- dia, and Dr. Robert H. Mohlen- brock, Southern Illinois University, for his critical reading of the entire manuscript.

Systematic Treatment

Key to the Illinois Genera of the Tribe Heliantheae (This key is admittedly partially technical so that each genus is keyed out only once. A strictly artificial key would result in many genera being keyed out in several places and would be greatly increased in size and complexity.)

1. Heads with ligulat.e or discoid flowers; corolla regularly developed.

2. Receptacle without chaff (merely bristly in Gaillardia).

3. Inner phyllaries united at base, glandular-dotted . 1. Dyssodia

3. Inner phyllaries free at base, not glandular-dotted.

4. Phyllaries herbaceous; heads radiate.

5. Plants with leafy stems.

6. Leaves decurrent on stem or, if not, then

linear-filiform . 2. Helenium

6. Leaves not decurrent on stem nor linear-filiform . 3. Gaillardia

5. Plants scapose . 4. Hymenoxys

4. Phyllaries petaloid, scarious; heads discoid . 5. Hymenopappus

2. Receptacle definitely with chaff.

7. Disc-flowers sterile (styles undivided, ovary reduced).

8. Cypselas* thick, scarcely flattened . 6. Polymnia

8. Cypselas compressed dorso-ventrally.

9. Flowers yellow, in large corymbose-panicled heads. .7. Silphium

9. Flowers white, in small corymbose heads . 8. Parthenium

7. Disc-flowers fertile (styles divided, ovary normal sized).

10. Cypselas turbinate, 5-angled . 9. Galinsoga

10. Cypselas flat, 4-angled, or, if 5-angled, then subterete and linear.

11. Ligules persistent on cypselas, chartaceous . 10. Heliopsis

11. Ligules promptly deciduous or absent.

12. Cypselas compressed dorso-ventrally (terete in Bidens beckii, an aquatic with filiform- dissected leaves); phyllaries dimorphic.

13. Pappus of 2 short teeth or awns, barbed upward or smooth, or a mere border, or absent; cypselas wing-margined (except C. tinctoria ) . 11. Coreopsis

13. Pappus of 2-6 awns or teeth, these barbed or hispid, usually retrorsely (rarely smooth or absent) ; cypselas not

wing-margined.

14. Cypselas beaked . 12. Cosmos

14. Cypselas not beaked . 13. Bidens

12. Cypselas scarcely flattened or sometimes compressed laterally; phyllaries not dimor¬ phic.

15. Heads discoid, appearing gray because

of black-tipped anthers . 14. Melanthera

Wunderlin — Illinois Flora

319

15. Heads radiate (inconspicuously in Eclip- ta), not gray.

16. Heads small, with short white ligules subequal to phyllaries; receptacle chaff bristleform; small weak an¬ nuals with short-stalked axillary heads . 15. Eclipta

16. Heads large, with yellow or pink to purple ligules much longer than phyllaries; receptacle chaff broader; stout perennials, biennials, or some¬ times annuals with peduncled or terminal heads

17. Receptacle conical or columnar (if conical, then leaves not de¬ current) .

18. Cypselas compressed later-

erally . 16. Ratibida

18. Cypselas 4-angled.

19. Flowers yellow (or mot¬ tled with brown) . 17. Rudbeckia

19. Flowers pink to purple

(in ours) . 18. Echinacea

17. Receptacle flat to convex (rarely conical in Verbesina).

20. Leaves decurrent on stem; cypselas compressed later¬ ally; pappus of 2-3 persis¬ tent awns . 19. Verbesina

20. Leaves not decurrent on stem; cypselas 3- to 4-an- gled; pappus of 2-4 cadu¬ cous scales . 20. Helianthus

1. Heads without ligulate flowers; pistillate flowers without corolla or with corolla

reduced to a tube or ring around base of style; staminate flowers with regularly developed corolla.

21. Heads all alike; pistillate flowers few, marginal; staminate

flowers many, central; involucre of few rounded phyllaries. .21. Iva

21. Heads of two kinds, pistillate with tuberculate or bur-like

involucre.

22. Staminate involucre with united phyllaries . 22. Ambrosia

22. Staminate involucre with distinct phyllaries . 23. Xanthium

^Characteristic fruit of the Compositae. Similar to an achene but bicarpellate rather than unicarpellate and formed from an inferior ovary with adherent floral tissues on outside wall.

1. DYSSODIA Cav., Anal. Cienc. Nat. 6:334. 1802. Willdenowa Cav., Ic. 1:61. 1791.

Boebera Willd., Sp. PI. 3:2125. 1804. Schlechtendalia Willd., Sp. PI. 3:2125.

1804, non Less., 1830.

Adenophyllum Pers., Syn. PI. 2:458. 1807. Thymophylla Lag., Gen. & Sp. Nov. 25. 1816.

Hymenatherum Cass., Bull. Soc. Philom. 1817:12. 1817.

Clomenocoma Cass., Diet. Sci. Nat. 9:416. 1817.

Lebetina Cass., Diet. Sci. Nat. 25:395 1822.

Rosilla Less., Syn. Gen. Comp. 245. 1832. Syncephalantha Barth, Ind. Sem. Hort.

Goett. 6.1836, ex Linnaea 12:80. 1838. Gnaphaliopsis DC., Prodr. 7:258. 1838. Lowellia Gray, Mem. Amer. Acad. II. 4:89. 1849.

Aciphyllaea (DC.) Gray, Mem. Amer. Acad. II. 4:91. 1849.

Comaclinium Scheidw. & Planch, ex Planch., FI. Seres 8:19. 1852.

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Transactions Illinois Academy of Science

Urbinella Greenman, Proc. Amer. Acad. Arts 39:117. 1903.

Gymnolciena (DC.) Rydb., N. Amer. FI. 34:160. 1915.

Boeberastrum (Gray) Rydb., N. Amer. FI. 34:161. 1915.

Trichaetolepis Rydb., N. Amer. FI. 34: 170. 1915.

Dysodiopsis (Gray) Rydb., N. Amer. FI. 34:170. 1915.

Annual or perennial herbs; leaves op¬ posite or alternate, entire to pinnately dissected, glandular-dotted. Heads sev¬ eral, terminal, radiate or rarely discoid; involucre in one or two series, conspicu¬ ously glandular-dotted, inner united at base or above; receptacle flat or nearly so, naked or nearly so; ray-flowers pistillate, fertile, ligule yellow or orange; disc-flow¬ ers numerous, perfect; anthers narrow at base; style-branches flattened, truncate, or with elongate pubescent appendages; pappus of 10-20 scales, each divided to middle or below into several bristles or entire. Cypselas narrow, substriate.

The genus Dyssodia consists of about 40 species native to the western hemis¬ phere. It is represented in Illinois by the following single species.

1. DYSSODIA PAPPUSA (Vent.) Hitchc., Trans. Acad. St. Louis 5:503. 1891.

Tagetes papposa Vent., Descr. Cels. pi. 36. 1802.

Boebera chrysanthemoides Willd., Sp. PI. 3:2125. 1804.

Tagets pumila Willd., Sp. PI. 3:2126. 1804, pro syn.

Boebera glandulosa (Cav.) Pers., Syn. PI.

2:459. 1807, nom nud.

Dyssodia chrysanthemoides (Willd.) Lag., Gen. et Sp. Nov. 29. 1816.

Dyssodia fastigiata DC., Prodr. 5:640.

1836, non D. fastigiata HBK., 1820. Dyssodia chrysanthemifolia Steud., Nom. Bot. II. 2:660. 1841.

Boebera papposa (Vent.) Rydb., in Bret- ton, Man. FI. N. States & Canada 1012. 1901.

Boebera ciliosa Rydb., N. Amer. FI. 34 :168. 1915.

Boebera roseata Rydb., N. Amer. FI. 34: 169. 1915.

Dyssodia ciliosa (Rydb.) Standi., Field Mus. Pub. Bot. 4:299. 1929.

Dyssodia roseata (Rydb.) Gentry., Los Pastizales de Durango 331. 1957.

Much branched, ill-scented annual; stems 0. 5-4.0 dm tall, puberulent to gla¬ brous; leaves opposite, 2-5 cm long, pin- natifid or bipinnatifid into linear or fili¬ form segments. Heads sessile or subsessile, numerous; involucre campanulate, 6-8 mm high, biseriate, outer phyllaries linear, subherbaceous, two-thirds or as long as

the wider more chartaceous inner, inner phyllaries united at base, with conspicu¬ ous elliptic glandular dots; ray-flowers few, ligules oval, inconspicuous, erect, up to 1.5 mm long; pappus scales divided to near base into 5-10 bristles, about 3 mm long. Cypselas subangled, compressed, about 3 mm long, pubescent. 2n = 26 (Smith, 1964).

Dyssodia papposa occurs from Ohio to Montana, south to Arizona and Louisiana and is adventive north and east of this range. It occurs infrequently along road¬ sides and in fields throughout Illinois (Fig. 1). It flowers from September to October.

Figure 1. Distribution of Dyssodia papposa in Illinois.

2. HELENIUM L., Sp. PI. 886. 1753. Helenia L., Gen. PI. ed. 5. 377. 1754. Brassavola Adans., Fam. 2:127. 1763. Actinea Juss., Ann. Mus. Paris 2:425. 1803.

Mesodetra Raf., FI. Ludov. 141. 1817. Leptopoda Nutt., Gen. N. Amer. PI. 2:174. 1818.

Leptophora Raf., Am. Mo. Mag. Crit. Rev. 4:195. 1819.

Tetrodus Cass., Diet. Sci. Nat. 55:264. 1828.

Dugaldia Cass., Diet. Sci. Nat. 55:270. 1828.

Wunderlin — Illinois Flora

321

Hecubaea DC., Prodr. 5:665. 1836. Amblyolepis DC., Prodr. 5:667. 1836. Leptocarpha Raf. ex Endl. Gen 1383. 1841, pro syn.

Oxylepis Benth., PI. Hartw. 87. 1841. Expeletiopsis Schultz-Bip. ex Benth. & Hook., Gen. 2:414. 1873.

Heleniastrum (Vaillant) Kuntze, Rev. Gen. 341. 1891.

Annual or perennial herbs; leaves alter¬ nate, impressed-punctuate, usually de¬ current. Heads corymbiform or solitary, radiate, rarely discoid: involucre in 2-3 series, subequal or the inner shorter, nar¬ row, herbaceous or subherbaceous, soon deflexed, outer sometimes joined at base; receptacle convex to ovoid or conic, naked; ray-flowers pistillate or sterile, yellow or occasionally purplish at base, ligule cuneate, 3- to 4-lobate, few; disc- flowers perfect, yellow or purple, numer¬ ous; anthers minutely auriculate or sagit¬ tate; style-branches flattened, tips di¬ lated, subtruncate, penicillate; pappus scarious, hyaline, often awn-tipped scales. Cypselas 4- to 5-angled, with as many in¬ termediate ribs, pubescent or glabrous.

The genus Helenium consists of about 40 species native to the western hemi¬ sphere with three species occurring in Illinois.

The pappus characters given for the species are variable. These often unusual variations are not considered to be of taxonomic importance.

Key to the Illinois Species of Helenium 1. Leaves linear to lin¬ ear-filiform, not de¬ current . 1. H. amarum

1. Leaves linear-lanceolate to ovate, de¬ current.

2. Disc depressed-glo¬ bose; disc-flowers

yellow . 2. H. autumnale

2. Disc globose; disc-

flowers purplish. . . 3. H. fiexuosum 1. HELENIUM AMARUM (Raf.) Rock, Rhodora 59:131. 1957.

Galardia amarum Raf., FI. Ludov. 69. 1817.

Helenium tenuifolium Nutt., Jour. Acad. Phila. 7:66. 1834.

Heleniastrum tenuifolium (Nutt.) Kuntze, Rev. Gen. 342. 1891.

Erect annuals; stems up to 5 cm tall, glabrous or nearly so; leaves linear to lin¬ ear-filiform, entire, 1-8 cm long, 1-2 mm wide, not decurrent, glabrous. Heads cor¬ ymbose; phyllaries linear, punctate, her¬ baceous, soon deflexed; disc depressed- globose, 0. 5-1.0 cm in diameter; ray- flowers 5-10, pistillate, yellow, ligule 0.5- 1.0 cm long, 3-logate; disc-flowers yellow; pappus ovate to obovate, hyaline, with awn as long as body. Cypselas about 1

mm long, hispid on angles. 2n = 30 (Turner & Ellison, 1960; Jackson, 1962).

Helenium amarum occurs from Virginia south to Florida, west to Texas, and north to Kansas and Missouri. It is occasionally introduced or adventive elsewhere. It is frequent in fields and waste ground in southern Illinois, extending north to Pike and Champaign counties (Fig. 2). It flowers from August to October.

Figure 2. Distribution of Helenium amarum in Illinois.

2. HELENIUM AUTUMNALE L., Sp.

PI. 886. 1753, non Gray, 1857. Helenium latifolium Mill., Gard. Diet. ed. 8. Helenium no. 2. 1768.

Helenia autumnalis (L.) Hill, Hort. Kew. 6. 1769.

Helenium pubescens Ait., Hort. Kew.

3:227. 1789, non H. & A., 1838. Helenium canaliculatum Lam., Jour. Hist. Nat. 2:213. 1792.

Helenia decurrens Moench, Meth. 598. 1794.

Helenium longifolium Smith, in Rees, Cycl. 17 : Helenium no. 2. 1811. Helenium pumilum Willd., Enum. Suppl. 60. 1813.

Helenium altissimum Link, Ind. Sem. Berol. 1840:21. 1840. nom. nud.

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Helenium commutation Link, Ind. Sem.

Berol. 1840:21. 1840, nom. nud. Helenium parviflorum Nutt., Trans.

Amer. Phil. Soc. II. 7:384. 1841. Helenium autumnale L. var. canalicula- tum (Lam.) T.&G., FI. N. Amer. 2:284. 1843.

Heleniastrum autumnale (L.) Kuntze, Rev. Gen. 342. 1891.

Heleniastrum parviflorum (Nutt.) Kuntze, Rev. Gen. 342. 1891.

Helenium autumnale L. var. pubescens (Ait.) Britton, Mem. Torr. Club 5:339. 1894.

Helenium altissimum Link ex Rydb., N.

Amer. FI. 34:126. 1915.

Helenium huronense Britton ex Rydb., N.

Amer. FI. 34:127. 1915, nom. nud. Helenium autumnale L. var. parviflorum (Nutt.) Fern., Rhodora 45:492. 1943. Erect perennials; stems up to 1.5 m tall, glabrous or finely strigose or puberu- lent; leaves linear-lanceolate to elliptical, acute, narrowed to sessile or subsessile base, decurrent along stem, 4-16 cm long, 0.5-5. 5 cm wide, serrate to subentire, gla¬ brous or occasionally puberulent. Heads corymbose; phyllaries lanceolate-subu¬ late, strigose or puberulent, soon deflexed; disc depressed-globose, 1-2 cm in diame¬ ter; ray-flowers 10-20, pistillate or occas- sionally neutral, yellow, ligule 0.5-1. 5 cm long, 3- to 4-lobate; disc-flowers yellow; pappus ovate to lanceolate, with awn up to 1 mm long. Cypselas about 1.5 mm long, hispid on ribs. 2n = 34 (Janaki- Ammal, 1945).

Helenium autumnale occurs from Que¬ bec, south to Florida, west to Arizona, and north to British Columbia. It is com¬ mon in wet meadows and along ditches, streams, and ponds throughout Illinois (Fig. 3). It flowers from August to Oc¬ tober.

A number of varieties are recognized by various authors. These are believed by the author to be variations within a polymorphic species and merit no taxo¬ nomic segregation.

3. HELENIUM FLEXUOSUM Raf., New FI. N. Amer. 81. 1838.

Helenium quadridentatum Hook., Comp.

Bot. Mag. 1:98. 1835, non Labill, 1792. Helenium dichotomum Raf., New FI. N. Amer. 81. 1838.

Helenium nudiflorum Nutt., Trans. Amer.

Phil. Soc. II. 7:384. 1841.

Helenium micranthum Nutt., Trans.

Amer. Phil. Soc. II. 7:385. 1841. Leptopoda brachypoda T.&G., FI. N. Amer. 2:388. 1842.

Heleninum purpureum Hale ex T.&G., FI. N. Amer. 2:388. 1842. pro syn. Leptopoda brachypoda T.&G., var. B T.& G., FI. N. Amer. 2:388. 1842.

Figure 3. Distribution of Helenium autumnale in Illinois.

Helenium atropurpureum Kth. & Bouche, Ind. Sem. Hort. Berol. Anno 1845, Collectorum 12. 1845.

Helenium atropurpureum Kth. & Bouche var. qrandicephalum Lamaire, Ill. Hort. 10:375. 1863.

Helenium brachypoda (T.& G.) A. Wood, Am. Bot. FI. 182. 1870.

Helenium seminariense Featherman, La.

Univ. Rep. 1870:74. 1871.

Helenium nudiflorum Nutt. var. purpurea (Hale ex T.&G.) Gray, Proc. Amer. Acad. Arts 9:203. 1871.

Heleniastrum nudflorum (Nutt.) Kuntze, Rev. Gen. 342. 1891.

Helenium polyphyllum Small, FI. S. E. U.S. 1291. 1903.

Helenium floridanum Fern., Rhodora 45: 494. 1943.

Helenium godfreyi Fern., Rhodora 45:494. 1943.

Erect perennials; stem up to 1 m tall, subpuberulent; leaves oblong to linear- lanceolate, entire or subentire, sessile, de¬ current along stem, 3-12 cm long, 0. 5-2.0 cm wide. Heads corymbose; phyllaries lanceolate to linear-lanceolate, puberu¬ lent, soon deflexed; disc globose, 6-14 mm in diameter; ray-flowers 10-20, neutral, yellow or sometimes purplish; pappus

Wunderlin — Illinois Flora

323

ovate to lanceolate, with awn up to 0.5 mm long. Cypselas about 1 mm long, his¬ pid. 2n = 28 (Turner, 1959; Jackson, 1962).

Helenium flexuosum occurs from New England, south to Florida, west to Texas, and north to Michigan. It is local along roadsides, in meadows, and in pastures in Illinois but is more common in the south¬ ern one-half of the state (Fig. 4). It flow¬ ers from June to September.

Figure 4. Distribution of Helenium flexuosum in Illinois.

3. GAILLARDIA Foug., Mem. Acad. Sci. Paris 1786:5. 1788.

Gaillarda Foug., Obs. Phys. 29:55. 1786. sine sp.

Galardia Lam., Encyc. 2:590. 1788. Calonnea Buchoz ex Lam., Encyc. 2:590. pro syn.

Virgilia L‘Her., Virgilia 1788.

Polatherus Raf., Amer. Mo. Mag. 2:268. 1818.

Guentheria Spreng., Syst. 3:356. 1826. Cerostylis Less., Syn. Comp. 239. 1832. Agassizia Gray & Engelm. ex Gray, Proc. Amer. Acad. Arts 1:49. 1847, non Chav., 1830.

Annual, biennial, or perennial herbs, rarely suffruticose at base. Leaves alter¬ nate or basal, entire to pinnatifid. Heads

radiate or discoid; phyllaries in 2-3 series, ovate to lanceolate, at least upper reflexed in fruit; receptacle convex to subglobose, alveolate, usually fimbrillate, fimbrillae soft and short conic to stiff and spine-like; ray-flowers usually neutral, often want¬ ing; ligules, if present, broad, cuneate or flabelliform, deeply 3-lobed, yellow and/or purple; disc-flowers bisexual and fertile, 5-lobate; anthers auriculate at base; style- branches with glabrous and short to his- pidulous and filiform appendages; pap¬ pus of about 6- to 10-awned scales. Cyp¬ selas broadly obpyramidal, wholly or partly covered by long stiff, ascending hairs.

The genus Gaillardia consists of about 12 species native to western North Amer¬ ica. The genus is represented in Illinois by G. pulchella Foug., an escape from cultiva¬ tion. Gaillardia aestivalis (Walt.) Rock (=G. lutea Greene; G. lanceolata Michx.) has been attributed to Illinois by Bid- dulph (1944) on the basis of a single speci¬ men collected by Otto Kuntze supposedly near Cairo, Alexander County. The Kuntze specimen deposited in the her¬ barium of the New York Botanical Gar¬ den has the following on its label: “G. lanceolata 2868, 9/9/74” (in one hand¬ writing) “2868, Cairo U. St.” (in a second handwriting). Kuntze (Rev. Gen. 339. 1891) states the following: “Gaillardia lanceolata Michx. U. St.: Cairo, Miss.” Thus there is no mention of Illinois either on the label of Kuntze’s specimen or in his publication although Biddulph spe¬ cifically cites it from Cairo, Alexander County, Illinois. If the species was once in Illinois it has not been collected since 1874 although on the other hand, it may never have been collected in Illinois at all but at some other location. In view of the uncertain occurrence of G. aestivalis in Illinois, the author has chosen not to include it in this treatment.

1. GAILLARDIA PULCHELLA Foug., Mem. Acad. Sci. Paris 1786:5. 1788. Gaillardia bicolor Lam., Encyc. 2:590.

1788, pro syn.

Virgilia heliodes L’Her., Virgilia. 1788. Gaillardia lobata Buckl., Prod. Acad. Phila.

1861:459. 1862.

Branched annual herbs; stems 2-6 dm tall; striate, short-hirsute with ascending hairs; lower leaves oblanceolate, 4-8 cm long, 0. 5-2.0 cm wide, bluntly toothed or lobed, short-petiolate, upper leaves ob¬ long-lanceolate, 2-6 cm long, 0.5-1. 5 cm wide, acute, base sessile, often somewhat clasping, densely hispid beneath, sparsely long pubescent above. Heads terminal, 3-6 cm wide; peduncles 5-15 cm long; phyllaries lanceolate, long-acuminate, her¬ baceous with chartaceous bases, hirsute

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and ciliate above; receptacle fimbrillae subulate, stiff, longer than achenes; ray- flowers neutral, ligules 1-2 cm long, 3- lobate, yellow with purple base or wholly purple; disc-flowers yellow below, purple above; pappus of lanceolate scales 5-6 mm long, gradually tapering into an awn equaling body. Cypselas 2. 0-2. 5 mm long, densely hirsute. 2n — 34 (Biddulph, 1944, Stoutamire, 1955, 1958, 1960); 2n = 36 (Biddulph, 1944).

Gciillardici pulchella occurs naturally in dry sandy prairies and openings from Colorado and New Mexico, east to Min¬ nesota, Nebraska, Missouri, and Louis¬ iana and as an escape along roadsides and in waste ground east to the Atlantic states. It occurs as an escape locally throughout Illinois (Fig. 5). It flowers from June to July.

Figure 5. Distribution of Gaillcirdia pulchella in Illinois.

4. HYMENOXYS Cass., Diet. Sci. Nat. 55:278. 1828.

Actinella Nutt., Gen. 2:113. 1818, in part, non Pers., 1807.

Picradenia Hook., FI. Bor.-Am. 1:317. 1833.

Phileozera Buckl., Proc. Acad. Phila. 1861:459. 1862.

Tetraneuris Greene, Pittona 3:265. 1898.

Rydbergia Greene, Pittonia 3:270. 1898. Macdougalia A. Heller, Bull. Torrey Club 25:629. 1898.

Plateilema (Gray) Cockerell, Bull. Torrey Club 31:462. 1904.

Aromatic annual or perennial herbs; leaves alternate or all basal (ours), entire or occasionally pinnately lobed. Heads solitary or few, radiate; involucres 2- to 3-seriate, subequal or slightly imbricate, appressed, herbaceous but often scarious margined; receptacle hemispherical or conic, naked; disc-flowers perfect, fertile; ray-flowers 10-20 pistillate, yellow; an¬ thers entire or sagittate at base; style branches flattened, truncate, penicillate; pappus of 5-12 hyaline, often aristate scales. Cypselas turbinate, mostly 5-an¬ gled, villous or sericeous.

The genus Hymenoxys consists of about 15 species native to the western hemi¬ sphere and is represented in Illinois by the following single taxon.

1. HYMENOXYS ACAULIS (Pursh) Parker var. GLABRA (Gray) Parker, Madrono 10:159. 1950.

Actinella scaposa (DC.) Nutt. var. glabra Gray, Man. Bot. ed. 5. 263. 1867. Actinella acaulis (Pursh) Nutt. var. glabra (Gray) Gray, Syn. FI. 2:345. 1884. Tetraneuris herbacea Greene, Pittonia 3: 268. 1898.

Actinea herbacea (Greene) B. L. Robins., Rhodora 10:68. 1908.

Actinea acaulis (Pursh) Spreng. var. gla¬ bra (Gray) Cronquist, Rhodora 47:403. 1945.

Perennial scapose herbs; stems 0.5-2. 5 cm tall; leaves narrowly to broadly ob- lanceolate, 1-8 cm long, 1.5-10.0 mm wide, villous when young, soon glabrate, strong¬ ly punctate. Head solitary, 3. 5-4.0 cm wide; involucre pubescent to subglabrate, 7-8 mm high; phyllaries broadly rounded; ligules 5-20 mm long, yellow; pappus ovate, acute or obtuse, about 2 mm long. Cypselas turbinate, about 3 mm long. Chromosome number unknown.

Hymenoxys acaulis var. glabra is very rare and occurs in dry, gravelly banks, stony fields, limestone hills, sandy fields and prairies, only in Mason and Will Counties, Illinois (Fig. 6), Ottawa County, Ohio, and southern Ontario, Canada. It flowers from May to July.

5. HYMENOPAPPUS L’Her., Hymenop. 1. 1788.

Rothia Lam., Jour. Nat. Hist. Paris 1:16. 1792, non Schreber, 1791, nec Bork- hausen, 1792; nec Pers., 1807.

Biennial or perennial scapose to leafy- stemmed herbs; leaves alternate, pinnati- fid or bipinnatifid to rarely simple, re¬ duced upwards. Heads several to numer-

Wanderlin — Illinois Flora

325

Figure 6. Distribution of Hymenoxys acaulis var. glabra in Illinois.

ous in a corymbiform panicle, radiate or discoid; involucre 2- to 3-seriate, inner usually with broad, scarious, petaloid, yellowish or whitish tips, outer herbace¬ ous; receptacle convex or nearly flat, naked or rarely with chaff; ray-flowers, when present, pistillate, fertile, ligules white; disc-flowers perfect, lobes reflexed, yellow or whitish (rarely purple) ; anthers cordate-sagittate at base; style branches flattened, with obtuse, papillose append¬ ages; pappus of 12-22 linear to ovate, ob¬ tuse, membranous or hyaline scales (rare¬ ly wanting). Cypselas turbinate, 4-angles, often striate.

The genus Hymenopappus consists of about 15 species native to North America and is represented in Illinois by the fol¬ lowing single species.

1. HYMENOPAPPUS SCABIOSAEUS L’Her., Hymenop. 1. 1788.

Rothia carolinensis Lam., Jour. Hist. Nat. Paris 1:17. 1792.

Hymenopappus laxiflorus L’Her., DC.

Prodr. 5:658. 1836, pro syn. Hymenopappus carolinensis (Lam.) Por¬ ter, Mem. Torrey Club 5:338. 1894. Biennial herbs; stems 3-15 dm tall, floccose-tomentose, becoming glabrate be¬ low, villous above; leaves pinnatifid or bipinnatifid, subpersistently floccose-to¬

mentose below, glabrate above, lower 8-25 cm long, 3-12 cm wide, reduced upwards. Heads several to numerous in open cor¬ ymbiform inflorescences, 7-12 mm wide; involucre, 7-15 mm high; phyllaries broad, scarious, petaloid white or yellowish; disc-flowers 5-lobed, lobes reflexed, half as long as tube or longer, tube stipitate- glandular, white or yellowish; pappus of 14-18 hyaline obovate scales, up to 1 mm long. Cypselas turbinate, 3. 5-5.0 mm long, 4-angled, striate, hirsute principally on angles. 2n = 34 (Raven & Kyhos, 1961).

Hymenopappus scabiosaeus occurs from Florida to Texas, north to South Caro¬ lina, Indiana, Illinois, and Kansas. It is rare in Illinois, known only from Cass, Iroquois, Kankakee, and Mason counties (Fig. 7) where it occurs in open sandy woods and prairies. It flowers from May to June.

6. POLYMNIA L., Sp. PI. 2:926. 1753.

Alymnia Neck., Elem. Bot. 1:31. 1790. Polyniastrum Lam., Tabl. Enc. t. 712.

1797.

Smallanthus Mackenz., in Small, Man.

S.E. FI. 1406. 1933.

Erect perennial herbs (ours) ; stems to 3 m tall; leaves opposite, pinnately or palmately veined, sessile or petiolate. Heads panicled corymbs, radiate, involu-

Figure 7. Distribution of Hymenopappus scabiosaeus in Illinois.

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Transactions Illinois Academy of Science

ere subfoliaceous, receptacle flat to con¬ vex; ray-flowers pistillate, ligule 2- 3-lo- bate to entire, sometimes wanting, white or yellow; disc-flowers staminate, yellow; pappus wanting. Cypselas obovoid or spherical, slightly flattened laterally or 3- to 5-angled.

The genus Polymnia consists of ap¬ proximately 20 species native to the western hemisphere with two species oc¬ curring in Illinois.

Key to the Illinois Species of Polymnia 1. Leaves pinnately lobed; cypselas 3-angled, not

striate . 1. P. canadensis

1. Leaves palmantely lobed; cypselas slightly flattened

laterally, striate . 2. P. uvedalia

1. POLYMNIA CANADENSIS L., Sp. PI. 2:926. 1753.

Polymnia variabilis Poir, Enc. Meth. 5:505. 1804.

Polymnia canadensis L. var. discoidea Gray, Gray’s Lessons in Bot. & Veg. Physio. 248. 1881.

Polymnia canadensis L. var. radiata Gray, Syn. FI. N. Am. 1:238. 1884.

Polymnia radiata (Gray) Small, FI. S.E. U.S. 1340. 1903.

Osteospermum canadense (L.) House, Bull.

N.Y. State Mus. 243:63. 1923.

Polymnia canadensis L. f. radiata (Gray) Fassett, Rhodora 34:96. 1932.

Erect perennial herbs; stems 0.5-1. 5 m tall, glandular-pubescent; lower leaves deeply pinnately lobed, up to 4 cm long, 3 cm wide, petiolate, upper triangular- ovate, entire to 3- to 5-lobed, smaller, petiolate. Heads in panicled corymbs, phyllaries 4-6, glandular-pubescent, sub- ovate; ray-flowers 4-6, white or pale yel¬ low, wanting or up to 1.5 cm long, 3- lobate, paleae ovate to ovate-lanceolate; disc-flowers white or pale yellow, paleae elliptic to oblanceolate, nearly equaling disc-flowers. Cypselas asymmetrically obovoid, 3-angled, not striate, 3-4 mm long, about 2-3 mm wide, dark brown to black. 2n =■ 30 (Wells, 1965).

Polymnia canadensis occurs from New England, Ontario, and Minnesota, south to Oklahoma, Louisiana, and Georgia. It is common in moist woods throughout Illinois (Fig. 8). It flowers from June to November.

Infraspecific categories based on ligule length of the ray-flowers have been named. The author, like Wells (1965), has chosen to treat these as variants of the species.

2. POLYMNIA UVEDALIA (L.) L., Sp. PI. ed. 2. 2:1303. 1764.

Figure 8. Distribution of Polymnia canadensis in Illinois.

Osteospermum uvedalia L., Sp. PI. 2:923. 1753.

Polymnia macrophylla Raf., FI. Ludov. 70. 1817.

Polymniastrum uvedalia (L.) Small, in Small and Carter, FI. Landcaster Co. 302. 1913.

Polymnia uvedalia (L.) L. var. genuina Blake, Rhodora 19:47. 1917.

Polymnia uvedalia (L.) L. var. densipilis Blake, Rhodora 19:48. 1917.

Polymnia uvedalia (L.) L. var. floridana Blake, Rhodora 19:48. 1917. Smallanthus uvedalia (L.) Mack., in Small, Man. S.E. FI. 1406. 1933.

Erect perennial herbs; stems up to 3 m tall, glabrous to densely glandular-pubes¬ cent; lower leaves deeply palmately 3- to 5-lobed, to 7 cm long, 4 cm wide, sessile or with winged petioles, upper leaves ovate, entire or toothed, sessile. Heads in panicled-corymbs, phyllaries 4-6, 20 mm long, ovate to ovate-lanceolate; ray- flowers 7-13, yellow, ligule about 3 cm long, paleae ovate, acuminate; disc-flow¬ ers yellow, paleae lanceolate. Cypselas asymmetrically obovoid, laterally com¬ pressed, striate, 5-6 mm long, 3-4 mm wide. 2n = 32 (Wells, 1965).

Polymnia uvedalis occurs from New

Wunderlin — Illinois Flora

327

England west to Missouri, south to Texas and Florida. It has recently been intro¬ duced into Bermuda (probably between 1883 and 1904 or 1905, cf. Wells, 1965). It is found in rich woods and is uncom¬ mon in southern Illinois (Fig. 9). It flowers from July to September.

Figure 9. Distribution of Polymnia uvedalia in Illinois.

The three varieties proposed by Blake (1917) and recognized by some authors are differentiated primarily on peduncle vestiture as well as geographical distribu¬ tion. Because of the broad overlap in their ranges and the lack of clear-cut distinc¬ tions among them, the author does not treat these variations as varietal entities

of the species. This treatment of the species is also now advocated by Wells (pers. comm.).

Literature Cited

Biddulph S. F. 1944. A revision of the genus Gaillardia. Res. Stud. St. Coll. Wash. 12:195-256.

Blake, S. F. 1917. Polymnia uvedalia and its varieties. Rhodora 19:46-48. Jackson, R. C. 1962. In documented chromosome numbers of plants. Man- drono 16:266-268.

Janaki-Ammal, E. K. in C. D. Darling¬ ton and E. K. Janaki-Ammal. 1945. Chromosome Atlas of Cultivated Plants. George Allen & Unwin Ltd., London.

Raven, P. H. and D. W. Kyhos. 1961. Chromosome numbers in Compositae. II Helenieae. Am. Jour. Bot. 48:842- 850.

Stoutamire, W. P. 1955. Cytological dif¬ ferentiation in Gaillardia pulchella. Am. Jour. Bot. 42:912-916.

_ 1958. Cytological varia¬ tion in Texas Gaillardias. Britonia 10: 97-102

_ 1960. The history of culti¬ vated Gaillardias. Baileya 8:13-17. Smith, E. B. 1964. Chromosome numbers of Kansas flowering plants I. Trans. Kans. Acad. Sci. 67:818-819.

Turner, B. L. 1959. Meiotic chromo¬ some counts for 12 species of Texas Compositae. Brittonia 11:173-177.

_ and W. L. Ellison. 1960.

Chromosome numbers in the Composi¬ tae. I. Meiotic chromosome counts for 25 species of Texas Compositae includ¬ ing 6 new generic reports. Texas Jour. Sci. 12:146-151.

Wells, J. R. 1965. A taxonomic study of Polymnia (Compositae). Brittonia 17: 144-150.

Wunderlin, R. P. 1968. Contributions to an Illinois Flora no. 2. Compositae I (Tribe Vernonieae). Trans. Ill. Acad. Sci. 61:132-138.

Manuscript received April 30, 1971.

CONTRIBUTIONS TO AN ILLINOIS FLORA No. 5. COMPOSITAE III. (TRIBE HELIANTHEAE, PART II— THE GENUS COREOPSIS)

R. P. WUNDERLIN

Department, of Biology, St. Louis University, St. Louis, Mo. 6310 f

Abstract. — Seven species of Coreopsis are reported for Illinois. Keys, descrip¬ tions, and distributional maps are pro¬ vided for each species.

This is the second part of a treat¬ ment of the tribe Heliantheae in Illinois. The first part, consisting of a key to the 23 Illinois genera and a treatment of the genera Dyssodia, Helenium, Gaillardia, Hymenoxys, Hymenopappus, and Polymnia, has been published previously (Wunder- lin, 1971). Treatments of the other genera will follow in subsequent publications.

Coreopsis consists of about 100 species native to North and South America, Hawaii, and Africa. Seven species occur in Illinois, three of which are adventive or escaped from cultivation.

Taxonomically the genus ap¬ proaches Bidens on one hand and Cosmos on the other, but it is gen¬ erally well demarcated as a genus in having wing-margined cypselas (rarely absent) and a pappus of two short teeth or awns, a short crown, or absent.

Many members of the genus are widely cultivated as ornamentals in temperate regions, several of which have escaped and have be¬ come naturalized.

The synonymy in this work es¬ sentially follows that of Sherff (1955).

The distributional maps are based upon specimens housed in the fol¬ lowing herbaria: Eastern Illinois University, the Field Museum of Natural History, the Illinois Nat¬ ural History Survey, the Illinois State Museum, the Missouri Bo¬ tanical Garden, Southern Illinois

University, the University of Illi¬ nois, and Western Illinois Univer¬ sity. The author is grateful to the curators of these herbaria for al¬ lowing him to study their speci¬ mens of Coreopsis. The author also gratefully acknowledges Dr. Robert H. Mohlenbrock, Southern Illinois University, for his critical reading of the manuscript.

Systematic Treatment

11. COREOPSIS L., Sp. PI. 907. 1753. Acispermum Neck., Elem. 1:34. 1790. Coreopsoides Moench, Meth. 594. 1794. Anacis Schrank, Denkschr. Akad. Mun- chen 5 (Math. Naturw.):5. 1817. Leachis Cass., Diet. Sci. Nat. 25:388. 1822.

Chrysomelea Tausch, Hort. Canal. (15). 1823.

Diplosastera Tausch, Hort. Canal. (15). 1823.

Calliopsis Reichenb., Ic. PI. Cult. pi. 70. 1823.

Chrysostemma Less., Syn. Gen. Comp. 227 1823

Electra DC., Prodr. 5:63. 1832.

Leptosyne DC., Prodr. 5:331. 1836. Agaristra DC., Prodr. 5:569. 1836. Epilepis Benth., PI. Hartw. 17. 1839. Tuckermannia Nutt., Trans. Amer. Phil. Soc. II. 7:363. 1841.

Pugiopappus A. Gray, in Torr., Pacif. Railr. Rep. 4:104. 1857.

Annual or perennial herbs; leaves op¬ posite or rarely alternate, simple, entire to deeply pinnately or palmately lobed or divided. Heads solitary or loosely cor¬ ymbose-paniculate, radiate; involucre double, outer foliaceous, usually spread¬ ing, inner wider, submembranous, ap- pressed; receptacle flat; palea membra¬ nous, striate, deciduous with fruit; ray- flowers usually neutral, ligule yellow, var¬ iegated, or rarely purple; disc-flowers bi¬ sexual, fertile, regular, 5-dentate, throat with annulus; anthers entire or subauricu- late; style-branches truncate, conic, or caudate; pappus 2-toothed, 2-awned, or absent. Cypselas (cf. Wunderlin, 1971) flat, obcompressed, often winged.

Key to the Illinois Species of Coreopsis

328

Wunderlin — Illinois Flora

329

1. Leaves undivided or rarely with 1 or 2 short lateral lobes.

2. Leaves mostly basal, linear to

oblanceolate . 1. C. lanceolata

2. Leaves produced to middle of stem

or higher, ovate to elliptic-lance¬ olate . 2. C. pnbescens

1. Leaves 3- to 5-lobed or divided.

3. Leaves sessile, deeply 3-lobed to or

below middle . 3. C. palmata

3. Leaves usually petiolate, divided into 3-5 segments.

4. Ligules of ray-flowers reddish- brown at base or throughout; disc-flowers reddish-brown.

5. Cypselas linear-oblong, wing¬ less; leaf segments linear to lin¬ ear-lanceolate. .4. C. tinctoria

5. Cypselas obovate, cartilagi- nous-margined; leaf segments lanceolate to elliptic-oblong to orbicular. . 5. C. basalis

4. Ligules of ray-flowers yellow; disc-flowers yellow or reddish- brown.

6. Leaf segments elliptic-lance¬

olate; cypselas 5-7 mm long; disc-flowers yellow or reddish- brown . 6. C. tripteris

6. Leaf segments linear to linear- lanceolate: cypselas 1-4 mm long; disc-flowers yellow

. 7. C. grandiflora

1. COREOPSIS LANCEOLATA L., Sp. PL 908. 1753.

Coreopsis crassifolia Ait., Hort. Kew.

3:253. 1787, non Sesse & Moc., 1894. Coreopsoides lanceolata (L.) Moench, Meth. 594. 1794.

Coreopsis lanceolata L. var. glabella Michx., FI. Bor.-Amer. 2:137. 1803. Coreopsis lanceolata L. var. villosa Michx., FI. Bor.-Amer. 2:137. 1803.

Leachia lanceolata (L.) Cass., Diet. Sci. Nat. 25:388. 1822.

Chrysomelea laneolata (L.) Tausch, Hort. Canal. (15). 1823.

Coreopsis oblongifolia Nutt., Jour. Acad. Phila. 7:76. 1834.

Coreopsis lanceolata L. var. succisaefolia DC., Prodr. 5:570. 1836.

Coreopsis lanceolata L. var. crassifolia (Ait.) Heynh., Nom. 1:219. 1840. Coreopsis lanceolata L. var. angustifolia T. & G., FI. N. Amer. 2:344. 1842. Coreopsis heterogyna Fern., Rhodora 40: 475. 1938.

Erect or ascending perennial herbs; stems 2-8 dm tall, scapiform, subangular, glabrous or pubescent; leaves opposite, simple or rarely with 1-2 small lateral lobes, linear to oblanceolate, 5-10 cm long, 0. 5-2.0 cm wide, lower long-petiolate, up¬ per sessile, pubescent or with ciliolate

bases only. Heads usually solitary, radi¬ ate, 3-6 cm wide; peduncles 2-4 cm long; outer phyllaries lanceolate to oblong- ovate, 4-8 mm long, inner ovate, 8-12 mm long; ray-flowers about 8, yellow, 1. 3-3.0 cm long, ligule obovate or cuneate, 3- lobate; palea linear-oblong with filiform tip, 4-6 mm long; disc-flowers yellow; style branches caudate; pappus with 2 small fimbriolate teeth. Cypselas obcom- pressed, orbicular, 2. 3-3.0 mm long, black calloused excrescences at ends, margin winged. 2n = 24, 48 (Bilquez, 1955); 2n — 26 (Turner, 1960).

Coreopsis lanceolata occurs naturally from Michigan and the Lake Superior region south to Florida and New Mexico and is escaped from cultivation and be¬ coming naturalized northeast to New England. It occurs locally in dry sandy or rocky soils throughout Illinois (Fig. 1), where it flowers from June to July.

Figure 1. Distribution of Coreopsis lanceolata in Illinois.

Coreopsis lanceolata var. villosa has been segregated by some authors. In Illinois it does not appear to have any distinct geo¬ graphical range and both varieties fre¬ quently grow together with intermediate forms present. It therefore is not recog¬ nized in this study.

330

Transactions Illinois Academy of Science

2. COREOPSIS PUBESCENS Ell., Bot.

S. C. & Ga. 2:441. 1823.

Coreopsis auriculata L. var. T. & G., FI. N. Amer. 2:343. 1842.

Coreopsis auriculata L. var. T. & G., FI.

N. Amer. 2:344. 1842, excl. syn. Coreopsis pubescens Ell. var. typica Sherff, Brittonia 6:341. 1948.

Erect perennial herbs; stems 5-13 dm tall, pubescent; leaves opposite, lower ovate to obovate, 2-5 cm long, 1-4 cm wide, petiolate, upper ovate to elliptic- lanceolate, entire or 3- to 5-parted, 5-10 cm long, 1-7 cm wide, sessile. Heads usu¬ ally solitary, radiate, 3-5 cm wide; pe¬ duncles 1-2 cm long; outer phyllaries lin¬ ear-lanceolate, 7-10 mm long, inner ovate, subequal; ray-flowers about 8, yellow, 1.0-2. 3 cm long, ligule cuneate to oblong- cuneate, 3-lobate; palea elongate-filli- form, 6-8 mm long; disc-flowers yellow; style-branches abruptly narrowed, linear- appendaged; pappus with 2 fimbriate teeth. Cypselas obcompressed, suborbicu- lar, 2. 8-3.0 mm long, black, glabrous or papillate, calloused excrescences at ends, margin winged. 2n = 28 (Snoad, 1952; Turner, 1960).

Coreopsis pubescens occurs naturally from Virginia, south to Florida, west to

Figure 2. Distribution of Coreopsis pubescens in Illinois.

Oklahoma, and north to Missouri and is adventive or escaped from cultivation elsewhere. It occurs locally in open woods, fields, and along roadsides in southern Illinois (Fig. 2). It flowers from June to September.

3. COREOPSIS PALMATA Nutt., Gen. 2:180. 1818.

Calliopsis palmata (Nutt.) Spreng., Syst. 3:611. 1826.

Coreopsis pauciflorci Lehm. ex Schlecht., Linnaea 10 (Litt.-Ber.) :76. 1836. Coreopsis praecox Fresen. ex Schlecht., Linnaea 10 (Litt.-Ber.) :76. 1836.

Erect perennial herbs; stems 5-9 dm tall, glabrous or hirsute at nodes; leaves opposite, principal palmately 3-parted into entire or irregularly lobed oblong or oblong-linear segments, 0.4-2. 5 cm long, 3-6 mm wide, sessile. Heads 1-3 (-6), radiate, 2. 5-6.0 cm wide; outer phyllaries linear-oblong, obtuse, margins scabrous- ciliolate; ray-flowers about 8, yellow, 1.5- 2.7 cm long, ligule oblong-obovate, slight¬ ly dentate; palea filiform, 6-7 mm long; disc-flowers yellow; style-branches nar-

Figure 3. Distribution of Coreopsis palmata in Illinois.

rowly conic; pappus 2-toothed. Cypselas obcompressed, elliptic-oblong, 5.0-6. 5 mm long, black, glabrous, narrowly winged. Chromosome number unknown.

Wunderlin — Illinois Flora

331

Coreopsis palmata occurs from Wiscon¬ sin and Manitoba south to Indiana, Mis¬ souri, and Oklahoma. It occurs in prairies, open woods, and along roadsides nearly throughout Illinois (Fig. 3). It flowers from June to July.

4. COREOPSIS TINCTORIA Nutt., Jour. Acad. Phila. 2:114. 1821. Calliopsis bicolor Reichenb., Ic. PI. Cult. pi. 70. 1823..

Diplosastera tinctoria (Nutt.) Tausch, Hort. Canal. (16). 1823.

Calliopsis tinctoria (Nutt.) DC., Prodr. 5:568. 1836.

Calliopsis tinctoria (Nutt.) DC. var. atro- purpurea Hook., Curtis’s Bot. Mag. 63:ph 3511. 1836.

Coreopsis bicolor Bosse ex Buch., Linnaea 25:630. 1853, nom. nud.

Coreopsis elegans Hort. ex Wieg., in Bailey, Stand. Cyc. Hort. ed. 2. 2:845. 1914, pro syn.

Coreopsis nigra Hort. ex Wieg., in Bailey, Stand. Cyc. Hort. ed. 2. 2:845. 1914, pro syn.

Coreopsis marmorata Hort. ex Wieg., in Bailey, Stand. Cyc. Hort. ed. 2. 2:845. 1914, pro syn.

Coreopsis tinctoria Nutt. var. nana Hort. ex Wieg., in Bailey, Stand. Cyc. Hort. ed. 2. 2:845. 1914.

Coreopsis radiata Hort. ex Wieg., in Bailey, Stand. Cyc. Hort. ed. 2. 2:846. 1914, non Mill., 1768.

Coreopsis tinctoria Nutt. f. atropurpurea (Hook.) Fern., Rhodora 44:477. 1942. Coreopsis tinctoria Nutt. var. atropurpurea Hook. f. tinctoria Sherff, Brittonia 6: 341. 1948.

Coreopsis tinctoria Nutt. var. atropurpurea Hook. f. atropurpurea Fern., ex Sherff, Brittonia 6:341. 1948.

Erect annual herbs; stems 6-12 dm tall, glabrous, subquadrangulate; leaves op¬ posite, subsessile, basal and lower pin- nately divided, segments linear, 5-10 cm long, 1-2 mm wide, upper entire. Heads numerous, subcorymbose, radiate, 1. 5-3.0 cm wide; peduncles slender, 4-10 cm long; outer phyllaries linear-oblong to triangu¬ lar, about 2 mm long, edges scarious, in¬ ner ovate, 5-7 cm long; ray-flowers 7-8, yellow with reddish-brown bases, 0.7-1. 5 cm long, ligule obovate, usually 3-lobate; palea subfiliform, 4. 0-4. 5 mm long; disc- flowers reddish-brown; style-branches ob¬ tuse; epappose. Cypselas obcompressed, linear-oblong, 1. 5-4.0 mm long, black, papillose below, wingless. 2n = 24 (Turn¬ er, 1960).

Coreopsis tinctoria occurs naturally in low ground from Minnesota, south to Louisiana, west to California, and north to Washington and is adventive or es¬ caped from cultivation east to the Atlan¬

tic coast. It occurs locally along roadsides, railroads, and in waste ground in Illinois as an adventive or escape (Fig. 4). It flowers from July to September.

Figure 4. Distribution of Coreopsis tinctoria in Illinois.

Coreopsis tinctoria f. atropurpurea is segregated by some authors. Since it in¬ tergrades with f. tinctoria and is a culti- gen its validity as a distinct taxon is in the opinion of the author not warranted. 5. COREOPSIS BASALIS (Otto & Dietr.) Blake, Proc. Amer. Acad. 51: 525. 1916.

Calliopsis basalis Otto & Dietr., Allg.

Gart. 3:329. 1835. (17 Oct.). Calliopsis drummondii D. Don, in Sweet, Brit. FI. Gard. II. pi. 315. 1835. (1 Dec.).

Coreopsis diversifolia Hook., Curtis’ Bot.

Mag. 63 :pl. 31+71+. 1836, excl. syn. Coreopsis picta Hort. ex Sieb. & Voss, in Vilmorin, Blumengartnerei ed. 3. 1: 487. 1894.

Coreopsis basalis (Otto & Dietr.) Blake var. typica Sherff, Brittonia 6:341. 1948.

Erect annual herbs; stems 2-4 dm tall, subglabrous to pubescent; leaves oppo¬ site, basal and lower 1- to 3-pinnate, seg¬ ments lanceolate to elliptic-oblong to or-

332

Transactions Illinois Academy of Science

bicular, 3-8 cm long, 2-4 cm wide; petioles

1- 5 cm long. Heads numerous, subcor- ymbose, radiate, 3-6 cm wide; peduncles slender, naked, 5-15 cm long; outer phyl- laries linear-lanceolate, 5-9 mm long, in¬ ner ovate, 6-10 mm long; ray-flowers about 8, yellow with reddish-brown bases, 1.3-2. 3 cm long, ligule cuneate-obovate,

2- to 3-lobate; palea linear, 6-10 mm long; disc-flowers reddish-brown; style- branches obtusely conic; epappose. Cyp- selas obcompressed, obovate, 1.4-1. 8 (-2.0) mm long, black, papillate, incurved mar¬ gins thickened and cartilaginous. 2n = 26 (Gelin, 1934).

Coreopsis basalis occurs naturally in dry rocky soils only in Texas although it occurs as an escape from cultivation else¬ where in the eastern United States. It is known in Illinois only from the following collection: LAKE CO.: Cedar Lake, Lake Villa, Fuller 13272 (ISM) (Fig. 5).

6. COREOPSIS TRIPTERIS L., Sp. PI. 908. 1753.

Anacis tripteris (L.) Shrank, Denkschr. Akad. Munchen 5 (Math. Naturw.):7. 1817.

Chrysostemma tripteris (L.) Less., Syn.

Gen. Comp. 227. 1832.

Coreopsis tripteris L. var. T. & G., FI.

N. Amer. 2:341. 1842.

Figure 5. Distribution of Coreopsis basalis in Illinois.

Coreopsis tripteris L. var. deamii Standi., Rhodora 32:33. 1930.

Coreopsis tripteris L. var. intercedens Standi., Rhodora 32:34. 1930.

Erect perennial herbs; stems 1-3 m tall, glabrous or rarely pubescent; leaves op¬ posite, principal ones pinnately 3- to 5- parted, segments elliptic-lanceolate, 3-10 cm long, 1-3 cm wide, petioles 0. 5-3.0 cm long, rameal leaves usually simple, ses¬ sile. Heads subcorymbose, radiate, 3-5 cm wide; peduncles 3-8 cm long; outer bracts oblong-linear, obtuse, 2-3 mm long, inner oblong-ovate, 4-6 mm long; ray-flowers 7-8, yellow, 1.2-2. 4 cm long, ligule ellip-

Figure 6. Distribution of Coreopsis tripteris in Illinois.

tic-oblong, subentire or dentate; palea filiform, purple-striate, 5-7 mm long; disc- flowers yellow to reddish-brown; style- branches caudate; epappose. Cypselas ob¬ compressed, cuneate, 5-7 mm long, brown¬ ish-black, glabrous, margin winged. 2n = 26 (Gelin, 1934).

Coreopsis tripteris occurs from Ontario to Wisconsin, south to Georgia, Louisiana, and Kansas and is escaped from cultiva¬ tion northeast to New England. It occurs frequently in open woods and along road¬ sides throughout Illinois (Fig. 6b 7. COREOPSIS GRANDIFLORA Hogg ex Sweet, Brit. FI. Gard. pi. 175. 1826.

Wunderlin — Illinois Flora

333

Coreopsis boykiniana Nutt., Trans. Amer.

Phil. Soc. II. 7:358. 1841.

Coreopsis heterophylla Nutt., Trans. Amer. Phil. Soc. II. 7:358. 1841, non Cav., 1796, nec Bertol., 1848.

Coreopsis grandiflora Hogg ex Sweet var. subinter grifolia T. & G., FI. N. Amer. 2:345. 1842.

Figure 7. Distribution of Coreopsis grandiflora in Illinois.

Erect or ascending perennial or rarely annual herbs; stems 3-6 dm tall, sub- glabrous; leaves opposite, basal and lower simple or irregularly divided, upper 3- to 5-parted, segments linear to linear-lan¬ ceolate, 5-10 cm long, 1-5 mm wide; peti¬ ole 0. 5-4.0 cm long. Heads usually soli¬

tary, radiate, 3-6 cm wide; peduncles slender, 1.0-1. 5 dm long; outer phyllaries lanceolate, subulate, 5-9 (-18) mm long, margins ciliate, inner ovate, subequal; ray-flowers about 8, yellow, 1.3-2. 5 cm long, ligule cuneate-obovate, 3-lobate; palea linear, 6-7 mm long; disc-flowers yellow; style-branches conspicuously cus¬ pidate; pappus small or wanting. Cyp- selas obcompressed, orbiculate, 1-4 mm long, black, papillate, usually with cal¬ loused excrescences at ends, margin winged. 2n = 26 (Gelin, 1934).

Coreopsis grandiflora occurs naturally in sandy or rocky prairies and thickets from Missouri to Kansas, south to New Mexico and Florida and is adventive or escaped from cultivation in the midwest and New England areas. It occurs locally along roadsides and in waste ground as an adventive or escape in Illinois (Fig. 7).

Literature Cited

Bilquez, D. fide C. D. Darlington and A. P. Wylie. 1955. Chromosome Atlas of Flowering Plants. George Allen & Unwin Ltd., London.

Gelin, 0. E. V. 1934. Embryologische und cytologische in Heliantheae - Co- reopsfdinae. Act. Hort. Ber. 11:99-128. Sherff, E. E., in E. E. Sherff and E. J. Alexander. 1955. Compositae - Heli¬ antheae - Coreopsidinae. N. Amer. FI. Ser. II. 2:4-40.

Snoad, B. 1952. in Chromosome counts of species and varieties of garden plants. John Innes Hort. Instit. 42:47-50. Turner, B. L. 1960. Meiotic chromosome numbers in Texas species of the genus Coreopsis (Compositae - Heliantheae). The Southwest. Natur. 5:12-15. Wunderlin, R. P. 1971. Contributions to an Illinois flora no. 4 Compositae II. (Tribe Heliantheae, part I - Dyssodia, Helenium,Gaillardia, Hymenoxys, Hy- menopappus, and Polymnia) . Trans. Ill. Acad. Sci. (In press).

Manuscript received April 30, 1971.

ARYL ACETOACETATES— INFRARED AND ULTRAVIOLET SPECTRA

FORREST J. FRANK, JOHN H. LONG AND TERRY K. REID

Illinois Wesleyan University , Bloomington, Illinois 61701, and Fontana Elementary School, Fontana, Wisconsin 53125

Abstract. — A series of aryl acetoace- tates has been prepared and their infra¬ red and ultraviolet spectra have been de¬ termined in order to study the effect of the aryl group upon the keto-enol spec¬ troscopic peak positions.

It is well known that the infrared spectra of beta-ketoesters in the carbonyl region reveal frequencies due to both the keto and enol forms. Although work has been reported correlating spectra of various alkyl acetoacetates and alpha-substituted acetoacetates (Bellamy, 1958), study of the infrared spectra of aryl acetoacetates has not been re¬ ported.

The enol form of beta-ketoesters absorbs ultraviolet radiation, and alkyl acetoacetates are reported to absorb in a narrow region around 240 nm (Korte and Wusten, 1961). It is surprising that almost no work has been reported concerning ultra¬ violet absorption of aryl acetoace¬ tates.

Methods and Materials

The aryl acetoacetates were pre¬ pared by the method of Zavialov (1965); the physical constants of reported esters agree with values reported by Lacey (1954).

Table 1 gives the data on four unreported esters. Elemental anal¬ yses were obtained by Galbraith

Laboratories, Knoxville, Tennessee.

Infrared spectra were run in spec¬ tral grade chloroform in 0.1mm cells on a Beckman IR8 grating spectrophotometer with an auxil¬ iary recorder used for scale expan¬ sion. In order to provide absorption maxima for calibration, the 1944 cm-1 peak of polystyrene was re¬ corded and, without stopping the scan, the polystyrene was replaced with the cells and the four peaks of interest were recorded. The cells were removed, again without stop¬ ping the scan, and replaced with polystyrene and the 1181 and 1154 cm-1 peaks were recorded. The above three polystyrene peaks were used for calibration; the error of mea¬ surement is estimated to be± 3cm-1.

UV spectra were run in absolute ethanol on a Beckman DB spec¬ trophotometer with an estimated error of ± 2nm.

Results and Discussion

Infrared spectra frequencies mea¬ sured in chloroform are shown in Table 2 and are arranged in de¬ creasing order of the high frequency bands of the ester carbonyl bond. Assignment of the frequencies to the ester carbonyl (1764-1734 cm-1), keto carbonyl (1720-1713 cm-1), enol ester carbonyl (1673-1630 cm-1) and

Table 1. Unreported Aryl Acetoacetates

Aryl Group

mp (°C)

Elemental Analysis

Calcd

Found

% c

% H

% c

% H

p-nitrophenyl

67-69

53.82

4.06

53.66

4.19

‘2,4,6-trimethylphenyl

60.5-62

70.89

7.32

70.76

7.34

alpha-phenyl-p-cresyl

54-56

75.82

6.36

75.82

6.12

2, 6-dimethoxy phenyl

64.5-66

60.50

5.92

60.40

6.02

334

Frank, Long and Reid — Acetoacetates

335

Table 2. Wavenumber Assignment (cm-1) of Keto and Enol Bands of Substituted Aryl Acetoacetates in Chloroform

Aryl Group

Ester

C = 0

Ketone

C = 0

Ester

C = 0

Enol C = C

a-Naphthyl

1764

1720

1649

1627

2,6-Dimethoxyphenyl

1764

1719

1673

1609

a-Phenyl-p-cresyl

1759

1723

1668

1629

2,4 ,6-Trimethylphenyl

1757

1723

1667

1630

p-Tolyl

1750

1718

1666

1614

B-Naphthyl

1741

1717

1659

1630

p-Nitrophenyl

1738

1715

1650

1616

Phenyl

1737

1716

1630

1603

p-Methoxyphenyl

1734

1713

1637

1603

enol carbon-carbon double bond (1630-1603 cm-1) is in accordance with the work of Rasmussen and Brattain (1949) and Leonard et al. (1952).

Frequencies due to the ester car¬ bonyl group vary over a range of 30 cm-1. The highest frequencies are exhibited by compounds pos¬ sessing ortho groups on the aryl group suggestive of a steric effect, however the one exception, alpha- phenyl-p-cresyl ester, argues against a steric effect. The keto carbonyl range of values is only 10 cm-1, not unexpected because of the distance between the keto and ester group¬ ings. The trend of the keto carbonyl frequencies, excluding the first two values, parallels that of the ester carbonyl although the significance is lessened because of the closeness to the estimated error of ± 3 cm-1 for each frequency. The enol ester carbonyl bands vary by the greatest amount, 43 cm-1, and parallel close¬ ly the ester carbonyl frequencies

with the glaring exception of alpha- naphthyl and the lesser exception of p-methoxyphenyl. Enol carbon- carbon double bond frequencies vary by 27 cm-1 and no parallel exists with any of the other three frequency types.

Although alkyl acetoacetates have ultraviolet maxima which, in alcoholic solution, vary little from 240 nm (Korte and Wusten, 1961), the aryl acetoacetates have maxima, as shown in Table 3, which exhibit a wide range of values. The shift toward 220 nm is interesting for it is near 210 nm that alpha-beta un¬ saturated esters absorb; addition of a hydroxy group on the beta car¬ bon gives the large bathochromic shift to about 240 nm for alkyl acetoacetates (Filler and Naqvi, 1963). All the aryl acetoacetates reported here absorb at a lesser wavelength than alkyl except for the 2,4,6-trimethylphenyl which has a maximum about equal to that of ethyl acetoacetate. Three of the

Table 3. Ultraviolet Absorption Maxima of Substituted Aryl Acetoacetates in Absolute Ethanol

Aryl Group

Maxima (nm)

am

2 ,4 ,6-Trimethylphenyl

243

1200

a-Naphthyl

236

2200

p-Nitrophenyl

232

760

B-Naphthyl

229

1300

p-Methoxyphenyl

228

6000

p-Tolyl

225

1600

Phenyl

223

840

2,6-Dimethoxyphenyl

220

2400

336

Transactions Illinois Academy of Science

esters are p-substituted phenyl com¬ pounds with the most electron with¬ drawing member having the highest value and decreasing for p-methoxy groups, although both are higher than the unsubstituted phenyl. The bulky 2,4,6-trimethylphenyl ester absorbs at the highest wavelength with the less bulky alpha-naphthyl ester following. A steric effect sug¬ gested by these two groups is countered by the effect of the 2,6- dimethoxyphenyl ester which has the lowest absorption maxima. However, the electronic effect of two o-methoxy groups may be important.

Acknowledgements

This paper was presented at the 62nd Annual Meeting of the Illinois State Academy of Science, Decatur, Illinois, April, 1969. Much of the material was taken from a thesis submitted by Terry K. Reid in partial fulfillment of the re¬ quirements for the M.S.T. degree at Illi¬ nois Wesleyan University. We wish to thank Illinois State University for the use of their infrared spectrometer.

This work was supported in part by

the National Science Foundation, Grant

GW-2716.

Literature Cited

Bellamy, L. J. 1958. The Infrared Spec¬ tra of Complex Molecules. John Wiley and Sons, Inc., New York pp. 184-5.

Filler, R. and S. M. Naqvi 1963. Fluo¬ rine — Containing beta-Dicarbonyl Compounds. Tetrahedron 19, 879.

Korte, F. and F. Wusten 1961. Uber den Enologehalt von Acetessigsaurees- tern mit Verschiedenen Esteralkyl- Komponenten, Ann., 61+7, 18.

Lacey, R., Derivatives of Acetoacetic Acid. Part VIII. The Synthesis of Coumarins from Aryl Acetoacetates, J. Chem. Soc. 854 (1954).

Leonard, N. J., H. S. Gutowsky, W. J. Middleton, and E. M. Petersen 1952. The Infrared Absorption Spectra of Cyclic -Ketoesters, J. Am. Chem. Soc., 71+, 4070-4073.

Rasmussen, R. S. and R. R. Brattain 1949. Infrared Spectra of Some Car¬ boxylic Acid Derivatives, J. Am. Chem. Soc., 71, 1073-1077.

Zavialov, S., V. Cunar, I. Mikhailo- pulo, and L. Ovechkina 1965. Low Temperature Acetoacetylation of Weakly Nucleophilic Compounds, Te¬ trahedron, 22, 2003-2008.

Manuscript received February 15, 1971.

METHODS OF EVALUATING THICKNESS AND TEXTURE OF GLACIAL TILL IN STUDIES OF GROUND-WATER RECHARGE

KEMAL PISKIN

Illinois State Geological Survey, Urbana, Illinois 61801

Abstract. — The thickness and vertical permeability of the till that overlies the glacial drift and shallow bedrock aquifers in northeastern Illinois were measured to determine their relation to the recharge rate of ground water. The till impedes vertical movement of the water. The average thickness of the till at main pump¬ ing centers was determined from maps and from cumulative thickness curves. The saturated thickness of the confining bed was estimated by subtracting depth to the assumed water table from the average thickness of the confining bed. Vertical permeability at the pumping centers was not measured directly but was calculated on the assumption that leakage and pumpage are approximately equal at the pumping centers.

Relative permeability was inferred from textural data obtained from hy¬ drometer analyses of subsurface samples from 52 test holes that penetrated the full thickness of the glacial drift. Textural differences of the confining bed varied regionally. Sand is more plentiful than clay roughly west of the Fox River, in¬ dicating relatively high permeabilities, whereas the reverse is true east of the river. The findings, supported by data derived from hydrologic analyses at the pumping centers, indicated thickness and vertical permeability of the till do affect ground-water recharge.

Glacial drift and shallow bedrock aquifers are a major source of ground water in the area of northeastern Illinois that includes Cook, DuPage, Kane, Lake McHenry, and Will Counties. Where glacial drift aq¬ uifers (sand and gravel) and shal¬ low bedrock aquifers (principally Silurian dolomite) are overlain by confining beds of glacial till, leaky artesian conditions exist. The till beds impede the vertical movement of ground water to the aquifer.

The withdrawal of ground water from these aquifers has produced a number of cones of depression. The vertical permeability of the con¬

fining beds as well as the hydro¬ geologic character of the aquifer are two of the factors that control the depth and shape of the cone of depression. In this report, the cone of depression is simply called the pumping center.

Several pumping centers were de¬ scribed and studied by Zeizel et al. (1962) and Prickett et al. (1964). The area of a pumping center is measured by drawing flow lines at right angles to the piezometric sur¬ face contours. It is rarely symmet¬ rically shaped because production wells are usually unevenly distrib¬ uted and are pumped at the same rates.

Figure 1 shows the areal distri¬ bution of the confining beds and possible sand and gravel deposits of the glacial drift (Piskin, 1963) in seven of these pumping centers. Working in northeastern Illinois, Walton (1960) calculated the quan¬ tity of leakage through a confining bed (the Maquoketa Group of Or¬ dovician age) into an aquifer by employing the equation:

P'

Qc = jpt ^ hAc

where Qc = leakage through the

confining bed (gpd)

P' = vertical permeability of the confining bed (gpd/ft2)

m' = thickness of the con¬ fining bed through which leakage occurs (ft)

Ac = area of the confining bed through which leakage occurs (ft2)

337

338

Transactions Illinois Academy of Science

WISCONSIN

Figure 1. Distribution of eastern Illinois.

glacial drift deposits in pumping centers in north-

INDIANA

Piskin — Glacial Till

339

Ah = difference between the head of the aqui¬ fer and the source bed above the con¬ fining bed (ft)

It can be applied to leakage through a confining layer of glacial till to evaluate quantitatively the effect of such a layer on ground-water re¬ charge to underlying aquifers.

Geology

The shallow bedrock in north¬ eastern Illinois consists mainly of rocks of the Maquoketa Group (Or¬ dovician) and the Alexandrian and Niagaran Series (Silurian). Quater¬ nary deposits, principally glacial drift, blanket the bedrock.

The Maquoketa Group underlies the glacial drift along the western margin of the study area. It con¬ sists of three formations, two shale formations and a middle formation that is argillaceous dolomite or lime¬ stone interbedded with shale. Dolo¬ mite is also present in the upper and lower shale formations, both of which are commonly fossiliferous and fine grained. The dolomitic mid¬ dle formation is part of the shallow bedrock aquifer system.

Silurian deposits directly under¬ lie the glacial drift in, approxi¬ mately, the eastern three-fourths of the study region, which is where the seven pumping centers used in the tests are located. The Alexan¬ drian Series (lower part of Silurian) consists of argillaceous to finely sandy, cherty, gray to light brown, finely crystalline dolomite; the over- lying Niagaran Series consists of white to light gray, cherty, silty, finely crystalline to medium-cry¬ stalline dolomite. The total thick¬ ness of the Silurian deposits ranges from 0 to 465 feet, increasing to¬ ward the southeast. The Silurian is the principal water-yielding unit of the shallow aquifer system in north¬ eastern Illinois.

The unconsolidated Quaternary deposits, overlying the Silurian and Maquoketa, resulted principally from the extensive continental gla¬ ciers and subsequent action by wind and running water; thus, most of the region is covered with thick drift. The drift varies in thickness from 0 to about 500 feet, depending on bedrock topography as well as the present land surface (Piskin and Bergstrom, 1967). In general, the drift thickens from south to north and from east to west; the thickest deposits, exceeding 250 feet, are restricted to the northern part of the study area.

The glacial drift consists prin¬ cipally of unsorted ice-laid rock de¬ bris (till), sorted meltwater deposits (glaciofluvial), and ancient lake-bed sediments (glaciolacustrine). The till found throughout the region is dense clayey silt to gravelly sand; it is most commonly fine grained. The till deposits are the principal confining layer for both the shallow bedrock and glacial drift aquifers. Glaciofluvial deposits of the area, in the form of terraces, outwash plains, and valley trains, are char¬ acteristically coarser in texture (sand and gravel) than the till and com¬ monly exhibit cross-bedding. These sand and gravel deposits are the second major source of shallow ground water in the region. Glaci¬ olacustrine deposits, generally the finest grained sediments, are found along Lake Michigan and in west¬ ern Will County, and, like the till, serve as a confining layer.

In the seven pumping centers tested, till deposits occur at various levels within the outwash and val¬ ley train deposits. Locally, they rest on the underlying Silurian do¬ lomite, where basal sand and gravel are absent. The areal relations of sand and gravel and till deposits for the seven pumping centers are shown in Figure 1. More detailed

340

Transactions Illinois Academy of Science

geologic information on bedrock formations is given by Suter et al. (1959), Zeizel et al. (1962), and Hughes, Kraatz, and Landon (1966).

Method of Evaluation

The equation was used to deter¬ mine vertical permeability of the confining beds at the seven pump¬ ing centers. In this report, the West Chicago pumping center, located in T. 39 and 40 N., R. 9 E., DuPage County, Illinois, is taken as an ex¬ ample. The area of influence of pro¬ duction wells was about 28.0 square miles in 1960 (Zeizel et al., 1962). The main aquifer at this center is the Silurian dolomite, which is re¬ charged by vertical leakage through the overlying glacial drift. The drift itself is recharged by precipitation. Because water levels suggest that recharge from leakage increases with pumpage, recharge to the Silurian dolomite aquifer in the West Chi¬ cago pumping center was assumed to be equal to the pumpage, which was estimated to be 1.8 million gal¬ lons per day (mgd). Zeizel et al. (1962) estimated the rate of re¬ charge to the Silurian dolomite in

1960 to be 64,000 gpd per square mile.

The map in Figure 2 shows the sand and gravel at the West Chi¬ cago site is at least 15 feet thick (Piskin, 1963). These deposits are divided into two groups, surficial deposits with less than 10 feet of fine-grained overburden, and buried deposits with more than 10 feet of fine-grained overburden, mainly till. Surficial and buried sand and gravel are assumed to be separated by till at least 10 feet thick. Surficial de¬ posits, which attain a maximum thickness of 50 feet, cover an area of approximately 4 square miles; buried deposits, ranging from 15 to 100 feet thick occupy an area of about 10 square miles. Cross sec¬ tions (Fig. 2) show that the present land surface approximates the to¬ pography of the bedrock surface, except in the north where a small valley in the Silurian dolomite has no present topographic expression.

At the West Chicago pumping center, confining beds of glacial till, which range in composition from yellow-gray-brown sandy silt through clayey silt and sand to

Figure 2. Geology of West Chicago pumping center.

Piskin — Glacial Till

341

slightly silty clayey sand, directly overlie the sand and gravel or, in some cases, Silurian dolomite (Fig. 2). The confining beds range from less than 25 to more than 75 feet thick (Fig. 3A), the thicker beds being located in the northeastern part of the pumping center. To rep¬ resent the average thickness of the confining bed at this pumping cen¬ ter, the ratio of the area between the various isopachs to the total area was plotted against the thick¬ ness of the confining bed. The 50th percentile point of area (Fig. 3B) was then taken as the average thickness of the confining bed in the pumping center. The depth of the water table was assumed to be 10 feet (T. A. Prickett, personal communication, 1969). The satura¬ ted thickness of this bed may be roughly determined by subtracting the depth to the water table from

the average thickness of the con¬ fining bed.

The vertical permeabilities of the confining beds in the West Chicago, Wheaton-Lombard-Glen Ellyn, and Downers Grove-Westmont-Hins- dale-Clarendon Hills pumping cen¬ ters were calculated by using the equation with the values for saturated thickness inserted. Head loss (Ah) was estimated by comput¬ ing the difference in elevation be¬ tween the head of the aquifer and that of the source bed above the confining bed. The other hydrogeo¬ logic data for these centers were taken from Zeizel et al. (1962). Ver¬ tical permeabilities of the confining beds at the remaining four pump¬ ing centers (La Grange, Chicago Heights, Woodstock, and Liberty- ville-Mundelein) were determined from data taken from Prickett et al. (1964). These results were compared

Figure 3. Thickness map (A) and graphical representation of average thickness (B) of confining beds in West Chicago pumping center.

342

Transactions Illinois Academy of Science

to the vertical permeabilities for the four pumping centers obtained by Prickett from analysis of pump tests; the two set of data compare favorably.

To evaluate the effect of texture upon vertical permeability, grain- size distributions of the confining beds were obtained by hydrometer analyses of subsurface samples from 52 precisely- planned test holes drilled in the six-county area. These

test holes, penetrating the entire thickness of glacial drift, were part of a study on water management coordinated by the Northeastern Illinois Metropolitan Area Plan¬ ning Commission (Hackett and Hughes, 1965). Textural analyses of the confining beds were averaged and plotted in a triangular diagram (Fig. 4) to show the distribution of the sand-silt-clay ratios. Results were then compared with the ver-

<$>

Average, all holes with high sand-low clay ratio

,0A# Test holes

□ 13 36

<D

Average, all holes with low sand-high clay ratio

Figure 4. Average composition of confining beds penetrated in test holes.

PisJcin-— Glacial Till

343

tical permeabilities of the various confining beds in the seven pump¬ ing centers.

Results and Discussion

Table 1 shows the thickness and characteristics of the confining beds in the seven pumping centers; at these sites, total drift thickness ranges from 10 feet in the La Grange area to approximately 300 feet in the Woodstock and Liberty ville- Mundelein areas. The table indi¬ cates that the average saturated thickness of the confining bed is higher in the Woodstock and Liber- tyville-Mundelein centers where the drift is thickest. Because the areas of thicker drift generally contain thicker deposits of confining till as well as sand and gravel, drift thick¬ ness directly influences the satura¬

ted thickness of the confining bed. These results are in agreement vith previously published data on the saturated thickness of the confining bed in the Chicago Heights, La Grange, Libertyville-Mundelein, and Woodstock pumping centers (Prickett et ah, 1964). The satura¬ ted thickness of the confining bed at the other three pumping centers (Table 1) was found to be reason¬ able (T. A. Prickett, personal com¬ munication, 1970).

Results of hydrometer analyses of the confining beds in the 52 test holes previously mentioned were obtained from standard grain-size ratios for sand (2.0 mm and 0.062 mm) silt (0.062 and 0.004 mm), and clay (< 0.004 mm). These figures indicate that, although the percent¬ age of sand ranges from 6 to 50

Table 1. Thickness and Characteristics of Confining Beds in Pumping Centers

Pumping Center

Average

Thickness

(ft)

Assumed

Saturated

Thickness

(ft)

Characteristics

Chicago Heights

44

34

Till, brown silty clay, clayey silt, and occasional gray silty clay with gravel and trace sand.

Downers Grove-

Westmont-Hinsdale-

Till, gray silty clay with trace

Clarendon Hills

58

48

of sandy silt and brown gravel.

La Grange

45

35

Till, brown to gray silty clay and gray clayey sand with silt.

Libertyville-

Mundelein

150

140

Till, gray clayey silt, silty clay with trace of sand and gravel; occasional gray clay and peb¬ bles.

West Chicago

52

42

Till, yellow-gray-brown sandy silt, clayey silt, and sand to clayey sand with trace of silt.

Wheaton-Lombard-

Glen Ellyn

45

35

Till, gray sandy silt, gray to brown clayey silt, and silty clay with occasional sand and gravel.

Woodstock

119

109

Till, gray to brown clayey sand with medium to coarse sandy gravel, and pinkish brown sandy silt with clayey gravel.

344

Transactions Illinois Academy of Science

percent and that of clay from 16 to 49 percent, the amount of silt is more or less consistent, averaging 45 percent. These ratios are depen¬ dent on the character of local tills within the glacial drift.

The percentage of coarse mate¬ rials is generally higher in the west¬ ern than the eastern part of the area, and the reverse is true for the fine materials. This relation is ap¬ parent in Figure 4, which was com¬ piled by plotting the average per¬ centage composition of the confin¬ ing bed at each test hole. Because of its relative consistency, the per¬ centage of silt was not considered. On the basis these percentages, three main groups were identified. The first, which falls at the left side of the diagram (the western part of the study area), is characterized by high sand (32 to 50 percent) and low clay (16 to 28 percent) ratios; a second group at the far right (the eastern part of the area) is charac¬ terized by low sand (6 to 24 percent) and high clay (16 to 43 percent) ratios (Fig. 4). These two divisions are separated by a third group, which is characterized by interme¬ diate ratios of sand (25 to 30 per¬ cent) and clay (22 to 32 percent). In addition, the average composi¬ tion of all holes with high sand and low clay and low sand and high clay percentages within the two groups were averaged. These total aver¬ ages are shown in Figure 4 by hexa¬ gons W and E, which represent the average sand-silt-clay ratios as 39: 39:22 and 17:49:34, in west and east groupings, respectively.

Figure 5 illustrates the regional distribution of the 52 test holes where subsurface data were ob¬ tained for the confining beds. The holes with high sand and low clay ratios generally occupy the area west of the Fox River, while the the reverse is true east of the river. Holes with intermediate ratios are

about equally distributed within the two groups. To compare varia¬ tions in vertical permeability with textural distribution, the vertical permeability of the confining bed at each of the seven pumping cen¬ ters was also included in Figure 5.

The vertical permeabilities ob¬ tained for the confining beds in the latter four pumping centers com¬ pares favorably with previously published data by Prickett et al. (1964). The vertical permeabilities in the various centers ranges from 0.3 x 10“2 gpd/ft2 to 1.3 x 10-2 gpd/ ft2. The Woodstock pumping cen¬ ter, which is the only one located west of the Fox River, had the highest vertical permeability in the region. The rest of the pumping cen¬ ters, located east of the Fox River, have comparatively low vertical permeabilities. These results ap¬ pear to correspond with the re¬ gional distribution of sand-clay ra¬ tios within the confining beds at the various pumping centers. The vertical permeabilities and clay ra¬ tios of the confining beds in the seven pumping centers are plotted in Figure 6 with a linear regression line drawn through these points. This relationship suggests that the high sand and low clay percentages coincide with the area of high ver¬ tical permeability (west of the Fox River), while low sand and high clay percentages can be observed in the area with low vertical perme¬ abilities (east of the Fox River).

Summary

The saturated thickness and ver¬ tical permeability of a glacial till overlying an aquifer has, under leaky artesian conditions, an effect on the recharge rate of the aquifer. Although this relation has long been inferred in this region, no previous experimental data have been ob¬ tained to support it. Data from sub¬ surface sampling programs and from

Piskin— Glacial Till

345

WISCONSIN

Figure 5. Distribution of test holes and pumping centers. Sand-silt-clay ratios for the test holes and vertical permeability of confining beds for the pumping centers.

INDIANA

346

Transactions Illinois Academy of Science

Figure 6. Comparison of vertical permeability with clay ratio of confining beds in 7 pumping centers.

cone analyses of selected pumping centers, both used in this study, are generally reliable. However, the vertical permeability differences among the seven pumping centers have a limited range, and greater variations might be expected in other areas. The relation between sand-clay ratios and vertical perme¬ abilities can be better established when additional pumping centers, especially west of the Fox River, are considered. Supporting studies from other areas would prove help¬ ful.

Literature Cited

Hackett, J. E., and G. M. Hughes. 1965. Controlled drilling program in northeastern Illinois. Illinois Geol. Sur¬ vey Environmental Geology Note 1, 5 pp.

Hughes, G. M., P. Kraatz, and R. A. Landon. 1966. Bedrock aquifers of northeastern Illinois. Illinois Geol. Sur¬ vey Circ. 406. 15 pp.

Piskin, K. 1963. Sand and gravel aqui¬ fers of northeast Illinois. Illinois Geol. Survey unpublished maps.

_ _ _ _ and R. E. Bergstrom.

1967. Glacial drift in Illinois: Thickness and character. Illinois Geol. Survey Circ. 416. 33 pp.

Prickett, T. A., L. R. Hoover, W. H. Baker, and R. T. Sasman. 1964. Ground-water development in several areas of northeastern Illinois. Illinois Water Survey Rept. Inv. 47. 93 pp. Suter, Max, R. E. Bergstrom, H. F. Smith, G. H. Emrich, W. C. Walton and T. E. Larson. 1959. Preliminary report on ground-water resources of the Chicago region, Illinois. Illinois Geol. Survey and Illinois Water Survey Coop. Ground-water Rept. 1. 89 pp.

Walton, W. C. 1960. Leaky artesian aquifer conditions in Illinois. Illinois Water Survey Rept. Inv. 39. 27 pp. Zeizel, A. J., W. C. Walton, R. T. Sas¬ man, and T. A. Prickett. 1962. Ground-water resources of Du Page County, Illinois. Illinois Geol. Survey and Illinois Water Survey Coop. Ground-water Rept. 2. 103 pp.

Manuscript received June 13, 1970.

THE PILEATE PORE FUNGI OF ILLINOIS

ROBERT W. SCHANZLE

University of Illinois, Department of Botany, Urbana, Illinois

Abstract. — A survey was made to de¬ termine the species of pileate pore fungi which occur in Illinois. Studies of litera¬ ture and herbarium specimens show that at least 92 species can be found in the state. Of these, 22 species have not pre¬ viously been listed for Illinois.

The Polyporaceae, or pore fungi, are those Basidiomycetes with a hymenium composed of pores with¬ in which basidia are borne. The Boletes, which are typically fleshy, are excluded from this family. This study was undertaken to determine the species of pileate polypores that are native to the state of Illinois. Specimens were examined from sev¬ eral herbaria in the state, including those of Eastern Illinois University, Illinois State University, the Uni¬ versity of Illinois, and the Field Museum of Natural History. A few specimens from Southern Illinois University were also studied.

In addition to actual identifica¬ tion of specimens, a survey of pre¬ vious literature concerning Illinois polypores was made. Two impor¬ tant works which list Illinois poly¬ pores are Moffatt (1909) and Over¬ holts (1953). Other listings are given in McDougall (1917 and 1919), and Graham (1944).

In the following list, those spe¬ cies previously cited for the state will be followed by the name of the author who first reported them. Those species of which specimens have not been examined will be followed by “sec.“ (secundum) and the earliest author. If a species has not previously been reported from the state, it will be preceded by an asterisk and a reference for the col¬ lection from which specimens were examined will be given.

Daedalea aesculi (Schw. ex Fr.) Murr.

Overholts (as Daedalea ambigua ) Daedalea confragosa Bolt, ex Fries Moffatt

Daedalea farinacea (Fries) Overh.

Sec. Overholts

*Daedalea quercina L. ex Fries University of Illinois collection Daedalea unicolor Bull, ex Fries Moffatt

Favolus alveolaris (D.C. ex Fries) Quel. Moffatt

*Favolus rhipidium (Berk.) Sacc. University of Illinois collection *Fomes annosus (Fries) Cooke University of Illinois collection

Fomes applanatus (Pers. ex Wallr.) Gill. Moffatt

Fomes conchatus (Pers. ex Fries) Gill. Overholts

Fomes everhartii (Ell. & Gall.) Von Schrenk & Spaulding Moffatt

*Fomes fomentarius (L. ex Fries) Kickx Field Museum of Natural History collection

Fomes fraxineus (Bull, ex Fries) Cooke Sec. Moffatt

Fomes fraxinophilus (Peck.) Sacc. Moffatt

Fomes fulvus (Scop, ex Fries) Gill. Moffatt

Fomes igniarius (L. ex Fries) Kickx Moffatt

*Fomes johnsonianus (Murr.) Lowe University of Illinois collection Fomes lobatus (Schw.) Cooke Overholts

Fomes ohiensis (Berk.) Murr.

Overholts

Fomes pini (Thore ex Fries) Karst. Owens

Fomes populinus (Schum. ex Fries) Cooke

Moffatt (as Fomes connatus )

Fomes ribis (Schum. ex Fries) Gill. Moffatt

*Fomes rimosus (Berk.) Cooke University of Illinois collection Fomes scutellatus (Schw.) Cooke Moffatt

Fomes viticola (Schw.) Lowe Sec. Overholts (as Fomes tenuis )

Lenzites betulina (L. ex Fries) Fries Overholts

*Lenzites sepiaria (Wulf. ex Fries) Fries University of Illinois collection Lenzites trabea (Pers. ex Fries) Fries Overholts

347

348

Transactions Illinois Academy of Science

*Polyporus abietinus Dicks, ex Fries Field Museum of Natural History collection

Polyporus adustus Willd. ex Fries Moffatt

*Polyporus albellus Peck University of Illinois collection Polyporus arcularius Batsch ex Fries Moffatt

Polyporus berkeleyi Fries Moffatt

*Polyporus betulinus Bull, ex Fries University of Illinois collection Polyporus biennis (Bull, ex Fries) Fries Sec. Overholts Polyporus biformis Fries Moffatt (as Polystictus pergamenus ) Polyporus brumalis Pers. ex Fries Moffatt

Polyporus chioneus Fries Sec. Moffatt

Polyporus cinnabarinus Jacq. ex Fries Moffatt

Polyporus cinnamomeus Jacq. ex Fries Moffatt

Polyporus compactus Overh.

Sec. Overholts

Polyporus conchifer (Schw.) Fries Moffatt

Polyporus cristatus Pers. ex Fries Moffatt (as Polyporus poripes ) Polyporus croceus Pers. ex Fries Overholts

*Polyporus curtisii Berk.

University of Illinois collection Polyporus cuticularis Bull, ex Fries Sec. Overholts *Polyporus delectans Peck University of Illinois collection Polyporus dichrous Fries Moffatt

Polyporus distortus Schw.

Sec. Moffatt

Polyporus dryadeus Pers. ex Fries McDougall

Polyporus dryophilus Berk.

Sec. Overholts

*Polyporus elegans Bull, ex Trog Field Museum of Natural History collection

*Polyporus focicola Berk. & Curt. Eastern Illinois University collection Polyporus frondosus Dicks, ex Fries Moffatt

Polyporus fumosus Pers. ex Fries Moffatt

Polyporus galactinus Berk.

Sec. Moffatt

Polyporus giganteus Pers. ex Fries McDougall

Polyporus gilvus (Schw.) Fries Moffatt

Polyporus glomeratus Peck Sec. Overholts

Polyporus graveolens (Schw.) Fries

Sec. Overholts

Polyporus hirsutus Wulf. ex Fries Moffatt

*Polyporus licnoides Mont.

Field Museum of Natural History collection

Polyporus lucidus Leys, ex Fries Moffatt (as Fomes lucidus )

Polyporus molliusculus Berk. & Curt. Moffatt (as Polystictus biformis ) Polyporus mutabilis Berk. & Curt.

Sec. Overholts

Polyporus pocula (Schw.) Berk. & Curt. University of Illinois collection *Polyporus perennis L. ex Fries Field Museum of Natural History collection

Polyporus picipes Fries Moffatt

Polyporus radiatus Sow. ex Fries Sec. Overholts Polyporus radicatus Schw.

Moffatt

Polyporus resinosus Schrad. ex Fries Moffatt

Polyporus robiniophilus (Murr.) Lloyd Moffatt

Polyporus rutilans (Pers.) Fries Moffatt

Polyporus sanguineus L. ex Fries Sec. Overholts Polyporus schweinitzii Fries Moffatt

Polyporus semipileatus Peck Sec. Overholts

Polyporus spraguei Berk. & Curt. Overholts

Polyporus squamosus Micheli ex Fries Sec. Overholts

Polyporus sulphureus Bull, ex Fries Moffatt

*Polyporus tephroleucus Fries Field Museum of Natural History collection

Polyporus tulipiferae (Schw.) Overh. Overholts

* Polyporus umbellatus Pers. ex Fries Collection of author (received from K. Andrew West of Southern Illinois University)

*Polyporus unicolor Fries University of Illinois collection Polyporus versicolor L. ex Fries Moffatt

*Trametes americana Overh.

Field Museum of Natural History collection (as Fomes odoratus)

Trametes hispida Bagl.

Moffatt (as Trametes peckii )

Trametes malicola Berk. & Curt.

Sec. Overholts

Trametes mollis (Sommerf.) Fries Sec. Overholts

Trametes rigida Berk. & Mont.

Moffatt

Schanzle — Pilate Pore Fungi

349

Trametes sepium Berk.

Overholts

Trametes serialis Fries Sec. Overholts *Trametes trogii Berk.

University of Illinois collection

Acknowledgements

The author wishes to thank Dr. Wesley C. Whiteside for his help and encourage¬ ment during the early stages of this study. Special thanks are due Dr. Donald P. Rogers for his guidance during the final stages and his excellent help with taxo- nomical problems and identifications. Thanks are also due the many people who lent specimens for study, especially Dr. Anthony E. Liberta, Dr. Patricio Ponce de Leon, and Mr. K. Andrew West.

Literature Cited

Graham, V. O. 1944. Mushrooms of the

Great Lakes region. Chicago Acad. Sci. Nat. Hist. Surv. Spec. Pub. 5.

McDougall, W. B. 1917. Some edible and poisonous mushrooms. Bull. Ill. State Lab. Nat. Hist. 6(7).

_ 1919. Some fungi that are

rare or have not previously been re¬ ported from Illinois. Trans. Ill. State Acad. Sci. 7:104-107.

Moffatt, W. S. 1909. The higher fungi of the Chicago region. Part 1 — the Hy- menomycetes. Chicago Acad. Sci. Nat. Hist. Surv. Bull. 7(1).

Owens, C. E. 1936. Studies on the wood- rotting fungus, Fomes pini. II. Cul¬ tural characteristics. Amer. Jour. Bot. 23:235-254.

Overholts, L. 0. 1953. The Polypora- ceae of the United States, Alaska, and Canada. Univ. Mich. Press, Ann Arbor.

Manuscript received May 21+, 1971.

A CENSUS OF MOULD SPORES IN THE ATMOSPHERE

HANSEL M. DeBARTOLO, JR.

Stritch School of Medicine, Loyola University, Maywood, Illinois

Abstract. — Using Aurora, Illinois as the research area this study determines the ecology of atmospheric moulds by the culture plate method. 2,480 colonies have been counted, made up as follows: Al¬ ter naria, 992; Penicillium, 562; Aspergil¬ lus, 258; Hormodendrum, 223 ; Fusarium, 186; Mucor, 67; Helminthosporium, 63; Yeast, 60; Pleospora, 21; Epicoccum, 13; miscellaneous, 105. In addition to the above genera, less common air borne spores were found of the genera Acre- monium, Botryosporium, Mortierella, Phoma, Monilia, Chaetomium, Macrospo- rium, Trichoderma and Pullaria.

This paper is concerned with eco¬ logical considerations of viable air borne fungus spores in Aurora, Il¬ linois. The literature abounds with references indicating that mould spores are of especial economic sig¬ nificance, owing chiefly to their di¬ sease-inciting proclivities.

Alternaria solani cause tuber rot in potatoes (Falsom and Bonde, 1925 ), Fusarium cause wilting of cab¬ bage (Armstrong and Armstrong, 1952), and tomatoes (Bohn and Tucker, 1940), Aspergillus niger cause crown rot of peanuts (Gibson 1953 a and b), cotton boll rot (Ray, 1946), black rot of onions (Venka- tarayan and Delvi, 1951), spoilage of dates (Bliss 1946, Almandel

1961) , grape rot (Gupta 1956), seed¬ ling blight (Leukel and Martin, 1943), bole rot (Wallace, 1952; Lock,

1962) , garlic rot (Mathur and Ma- thur, 1958), stem rot (Natour and Miller, 1960), root rot (Alvarez and Diaz, 1949) and many other dis¬ eases (Christenson 1962, Durbin 1959).

In man asthma has been shown to be caused by Aspergillus (Bern- ton, 1930) by Hormodendrum (Cobe, 1932) by yeast (Taub, 1932) and by Alternaria by several workers (Hop¬

kins, et al. 1930; Underwood, 1938 and 1941; Harris, 1939; Chobot, et al. 1940; and Durham, 1937). Con¬ junctivitis has been shown caused by Alternaria and Hormodendrum (Simon, 1938). Aspergillus also has been shown to cause respiratory infection (Hertzog, et al., 1949; Orie, et al., 1960), cardiovascular lesions (Merchant, et al. 1958; Ha- dorn, 1960), skin lesions (Frank and Alton, 1933; Sartory & Sartory, 1945), eye infections (Fine, 1962) and generalized infections (Cawely, 1947, Grcevic and Matthews, 1959).

Pady (1962) and others (Kramer et al., 1963) conclude that the knowl¬ edge of numbers and types of viable air spora is greatly needed, yet present spore sampling techniques give no information on the viability of the spore load. Furthermore some genera, particularly those like Peni- cillum and Aspergillus, are identi¬ fiable only by culture. This study employs nutrient plates to deter¬ mine the ecology of viable air spora.

Materials and Methods

During June and July, 1968, Petri dishes were exposed at several locations. The culture media used in this investiga¬ tion were as follows:

1. Sabouraud Dextrose Agar TM-MFG slants received from Hy¬ land Div. Travenol Laboratories Los Angeles, California, USA

Lot 6285E8 pH - 5.6

2. Mycology Agar

TM-MFG slants received from Hy¬ land Div. Travenol Laboratories Los Angeles, California, USA List #56-200 Lot 6200-D9

3. Potato Dextrose Agar

Potatoes . 300 gm

Dextrose . 10 gm

Agar . 15 gm

H20 . to make 1 liter

Lactic Acid . to make pH 4.5

350

De Bartolo — Mould Spores in the Atmosphere

351

4. Water Agar

Agar . 25 gm

H20 . to make 1 liter

Lactic Acid . to make pH 3.5

Media No. 1 and 2 were tried at the out¬ set of the experiment. Since medium No. 1 was found to produce higher rates of growth and larger numbers of colonies, medium No. 1 was used throughout June. There was no appreciable difference in the range of organisms yielded by medium No. 1 and medium No. 3, yet medium No. 3 was found to produce higher rates of growth. Therefore, medium No. 3 was used throughout the remainder of the ex¬ posure trials. Medium No. 4 was found to be best suited to accept transfer of fast growing colonies.

After exposure the plates were sealed with cellulose tape to prevent excessive drying and contamination. The cultures were then incubated in a dark chamber at room temperatures which varied from 65-80°F. The initial incubation period ranged from five to twelve days. Each different colony was transferred by flamed needle loop to a sterile plate and incu¬ bated for further observation.

In an effort to provide an ecological survey of the air borne fungi, it appeared wholly essential to classify the localized fungi according to recognized criteria. The majority of the genera were identi¬ fied at 100X magnification with the use of an illustrated key (Barnett, 1960). In questionable cases, a higher magnification was used for examination of finer, struc¬ tural details, and frequently bits of colony were removed with a needle loop and examined under a cover slip as a moist preparation. This technique frequently disclosed spores when they could not be seen previously.

A daily survey was undertaken for a month so as to increase the breadth of the study for the total effectiveness of obtain¬ ing conclusions of significance and re¬ peatability. The exposure station was located in a position not immediately flanked by a taller structure, on a flat roof in a building in Aurora, Illinois at an elevation of 20 feet, 764 feet above sea level. Beginning August 1, 1968, four 90 mm Petri dishes were exposed consecu¬ tively for fifteen minutes daily between 7:00 p.m. and 8:00 p.m. for a period of thirty days (excluding August 3rd and 4th). Temperature was recorded at time of exposure and reported together with daily high and low values.

Results

The most abundant, identifiable air borne spores belong to the gen¬

era Alternaria, Penicillium, Asper¬ gillus, Hormodendrum ( Cladospori - um), Fusarium, Mucor, and Hel- minthosporium, Pleospora and Epi- coccum (Figures 1, 2, 3).

Of the 2480 colonies included in the survey, 2325 were placed in these nine genera (Figure 4). In ad¬ dition to the above genera, less com¬ mon air borne spores were found of the genera Acremonium, Botryo- sporium, Mortierella, Phoma, Mo¬ nilia, Chaetomium, Macrosporium, Trichoderma and Pullularia.

Figure 1. Fungus spore colonies at Aurora, Illinois. Each record represents the number of colonies recorded from 4, 90mm Petri plates exposed consecutively for 15 minutes daily from August 1, 1968 through August 31, 1968.

Discussion

Since many fungus spores are ca¬ pable of inducing respiratory illness, it would be interesting to postulate how many spores a person might

352

Transactions Illinois Academy of Science

Figure 2. Fungus spore colonies at Aurora, Illinois. Each record represents the number of colonies recorded from 4, 90mm Petri plates exposed consecutively for 15 minutes daily from August 1, 1968 through August 31, 1968.

inhale during a specific day. For example let us take Alter naria at Aurora, Illinois on August 17, 1968. By consulting our data (Table 1) for Alter naria at Aurora on the date specified we find 116 colonies per Petri plate (90 mm) per hour, or at least 37 spores per cm. per 24 hours. Assuming each colony was started by at least one spore, (however, several spores may have been clumped and this number may be much greater), we can now obtain the number of spores per cubic yard of air by multiplying by 14.90, the conversion factor for Alternaria (Durham, 1946). This gives 551 spores per cubic yard of air. Since an average adult male has a lung capacity (tidal air) of approximately 500 cubic centimeters, and breathes between sixteen and eighteen times per minute, he would inhale approx¬ imately 12,240,000 cubic centime¬

ters (12.24 cubic centimeters or 16 cubic yards) of air in a 24-hour pe¬ riod. Since there are 551 spores in each cubic yard of air, he would inhale approximately 8816 Alter¬ naria spores in a 24-hour period.

Figure 3. Fungus spore colonies at Aurora, Illinois. Each record represents the number of colonies recorded from 4, 90mm Petri plates exposed consecutively for 15 minutes daily from August 1, 1968 through August 31, 1968.

Basic weather data were obtained through Dr. Clarence F. Smith, Professor of Physics, Aurora Col¬ lege, and an attempt was made to correlate the prevalence of fungus spore colonies with weather data. It was found that the initial rain¬ fall was heavily laden with fungus spores. The data show that the number of genera increases with rain cloud altitudes. (Table 3). It was also observed that the perime¬ ters of pits formed by raindrops

Table 1. Fungus spore colonies at Aurora, Illinois. Each record represents the number of colonies recorded from 4, 90 mm Petri plates exposed consecutively for 15 minutes daily from August 1, 1968 through August 31, 1968.

De Bartolo — Mould Spores in the Atmosphere 353

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354

Transactions Illinois Academy of Science

Figure 4. Fungus spore colonies at Aurora, Illinois. Each record represents the number of colonies recorded from 4, 90mm Petri plates exposed consecutively for 15 minutes daily from August 1, 1968 through August 31, 1968.

were covered by colonies of fungi. This suggests that a raindrop col¬ lects spores as it falls through air. Although the colonies were too nu¬ merous to be counted, observations of the culture plates showed greater

numbers of colonies when they were derived from raindrops originating at high rather than from low alti¬ tudes.

In general, the number of fungus spore colonies was highest during the beginning of the rain showers but were few at the end. Even though the air tends to be washed free of fungus spores during a pro¬ longed rain, there is no assurance that a distant mass of air with its attendant high concentration of spores will not invade the area soon after the storm has passed.

Hamilton (1959) recorded the temperature at which different spore types were in the air in maximum numbers. She reported the optimum temperatures within a 4°F range for each spore category, and construc¬ ted graphs showing the effect of temperature on spore counts. This effect was not readily observed in this investigation. For example, on August 17th, the highest count of Alternaria was recorded (Table 1) at temperatures ranging from 68- 66°F (Table 2). On August 23rd, the next highest count of Alternaria was recorded at temperatures rang¬ ing from 88-89°F. In fact, exact cor¬ relation between temperatures and spore incidence is not readily evi¬ dent for any genus in this investi¬ gation (Figures 1-6). Perhaps local effluvia differ and thus the irre-

Table 2. Data collected from culture plates exposed during rain showers

on the dates given.

August 10th (Precipitation 0.86'') clouds altitude 500 ft.

August 1 7th ( Precipitation 3.83") clouds altitude 21,000 ft.

Alternaria . 27

Aspergillus . 4

Penicillium . 10

Unidentified . 1

Indistinguishable . over 400

Alternaria . 116

Aspergillus . 17

Epicoccum . 5

Fusarium . 2

Helminthosporium . 3

Hormodendrum . 8

Penicillium . 10

Pleospora . 7

Yeast like . 6

Unidentified . 12

Indistinguishable . over 400

De Bartolo— Mould Spores in the Atmosphere

355

Figure 5. High and low temperatures recorded at Aurora, Illinois, daily from August 1, 1968 through August 31, 1968.

concilable disparity between these findings and those of Hamilton.

It was observed (Table 3) that mowing of grass produces a great and immediate local increase in fungi spore count. This is evidence that human activity and local flora may play a part in affecting atmos¬

pheric spore concentration. Lacey (1962) has shown that local flora may influence air spora.

The study has produced informa¬ tion which is representative of the viable air borne fungus spore con¬ tent of the air (Figures 1-4). It

Figure 6. 7:00 P.M. and 8:00 P.M.

temperatures recorded at Aurora, Illinois, daily from August 1, 1968 through Au¬ gust 31, 1968.

Table 3. Data collected from culture plates exposed one foot above the ground in the center of a 1/4 acre lawn before and after mowing.

August 30th

Before mowing.

(One plate exposed 15 min.)

August 30th

After mowing.

(One plate exposed 15 min.)

Alternaria . 7

Aspergillus . 2

Hormodendrum . 3

Fusarium . 1

Unknown . 7

Alternaria . 34

Aspergillus . 11

Hormodendrum . 51

Helminthosporium . 5

Fusarium . 15

Penicillium . 21

Unknown . 2

356

Transactions Illinois Academy of Science

seems very likely that natural se¬ lection has tended to limit the spores transported by air. Most of the air borne fungi identified in this inves¬ tigation are those that have special adaptations for aerial dissemina¬ tion. Usually they are somewhat thick walled or are able to resist dessication, are prolific sporulators, are commonly found in nature, and have some of the widest host ranges.

Conclusions

The results of the nutrient plate study reveal that certain viable fungi are present in the air through¬ out at least a 3-month period but vary in quantity. It is evident that the fungus spore content of the at¬ mosphere at any given time may originate from unknown or local sources. The fungus spore shower records point out that the source may be from unknown altitudes.

Acknowledgements

The author is indebted to Dr. and Mrs. H. M. DeBartolo, Dr. J. T. Velardo, Dr. C. F. Smith, and Miss Michele DeBartolo for their generous assistance.

References

Almandel, A. R., 1961, Chemical Con¬ trol of Spoilage of Dates, Dissertation Abstr., 21 :1967.

Alvarez Garcia, L. A., and M. A. Diaz, 1949, Aspergillus Root Stalk Rot of Sanseviera, ( Sanseviera Laurentii, Wil- dem.) J. Agr. Univ. Puerto Rico, 33: 35. Armstrong, G. M. and J. K. Armstrong, 1952, Physiological Races of Fusarium Causing Inlets of the Cruciferae, Phy¬ topath., 1+2: 255.

Barnett, H. L., 1960, Illustrated Genera of Imperfect Fungi, Burgess Publishing Company.

Bernton, H. S., 1930, Asthma Due to a Mold — Aspergillusfumigatus, J.A.M.A., 95:189.

Bliss, D. E., 1946, The Use of Fungi¬ cides Against Spoilage in Dates, Rept. Ann. Date Growers’ Inst.

Bohn, G. W., C. M. Tucker, 1940, Stud¬ ies on Fusarium Wilt of Tomato, M.D. Arg. Exp. Sta. Res. Bull., 5:11. Cawely, E. P., 1947, Aspergillosis and the Aspergilli, Arch. Internal Med., 80:423.

Chobot, R., H. Dundy, and N. Schaf¬ fer, 1940, Relationship of Mold Re¬ actions to Clinical Symptoms, J. Al¬ lergy, 12:46.

Christensen, C. M., 1962, Invasion of Stored Wheat by Aspergillus ochraceus, Cereal Chem., 59:100.

Cobe, H. M., 1932, Asthma Due to Mold: Hypersensitivity due to Cladosporium fulvum, Cooke: A Case Report, J. Al¬ lergy, 3:389.

Durbin, R. D., 1959, The Possible Re¬ lationship between Aspergillus flaous and Alunism in Citrus, Plant Disease Reptr., 1+3: 922.

Durham, O. C., 1937, Incidence of Air Borne Fungus Spores: I. Alternaria, J. Allergy, 8:480.

_ , 1946, The Volumetric In¬ cidence of Atmospheric Allergens. IV. A Proposed Standard Method of Grav¬ ity Sampling, Counting, and Volumet¬ ric Interpolation of Results, Journal of Allergy, 17: 79.

Falsom, D., and R. Bonde, 1925, Alter¬ naria solani and Cause of Tuber Rot in Potatoes, Phytopath., 15:282.

Feinberg, S. M., and H. T. Little, 1935, Studies on the Relation of Microorgan¬ isms to Allergy, J. Allergy, 6:564.

Fine, B. S., 1962, Intraocular Mycotic Infections, Tab Invest., 11:1161.

Frank, L., and O. M. Alton, 1933, As¬ pergillosis, Case of Postoperative Skin Infection, J.A.M.A., 100:2007.

Gibson, I. A. S., 1953 a., Crown Rot, A Seedling Disease of Groundnuts Caused by Aspergillus niger, Brit. Mycol. Soc. Trans., 56:198.

_ , 1953b., Seedling Disease

of Groundnuts Caused by Aspergillus niger, Brit. Mycol. Sol. Trans., 36:324.

Grcevic, N., and W. F. Matthews, 1959, Pathologic Changes in Acute Dis¬ seminated Aspergillosis, Particularly Involvement of the Central Nervous System, Am. J. Clin. Path., 32: 536.

Gupta, S. L., 1956, Occurrence of Asper¬ gillus carbonarius (Bainer) Causing Grape Rot in India, Sci. and Culture, 22: 167.

Hadorn, W., 1960, Aorten Ruptur Durch Aspergillus Infektion Nach Operation Eines Aorten-Stanose, Schweiz, Med. Wochschr., 90:929.

Hamilton, E. D., 1959. Studies on the Air-Spora, Acta Allerg. Kbh., 15:143.

Harris, L. H., 1939, Allergy to Grain Dusts and Smuts, J. Allergy, 10:327.

Hertzog, A. J., T. S. Smith, and M. Goblin, 1949, Acute Pulmonary As¬ pergillosis Report of a Case, Pediatrics, 4:331.

De Bartolo — Mould Spores in the Atmosphere

357

Hopkins, J. G., R. W. Benham, and B. M. Kesten, 1930, Asthma Due to a Fungus — Alternaria, J.A.M.A., 91+:6. Kramer, C. L., S. M. Pady, and B. J. Wiley, 1963, Kansas Aeromycology XIII: Durnal studies 1959-1960, My¬ cologies., 55:380.

Lacey, M. E., 1962, The Summer Air Spora of Two Contrasting Adjacent Rural Areas, J. Gen. Microbiol., 29:485. Leukel, R. W., and J. H. Martin, 1943, Seed Rot and Seedling Blight of Sor¬ ghum, U.S. Department Agr. Tech. Bull. No. 839.

Lock, G. W., 1962, Sisal : Twenty-five years Sisal Research, Longamous, Lon¬ don.

Mathur, R. L., and B. L. Mathur, 1958, Black Mould of Garlic, Allium sativum Sci, & Culture, 23: 372.

Merchant, R. K., D. B. Louria, P. H. Greisler, J. H. Edgcomb, and J. Putz., 1958, Fungal Endocarditis: Re¬ view of the Literature and Report of Three Cases, Am. Int. Med., 1+8: 242. Nataur, R. M., and H. N. Miller, 1960, Stem Rot of Dracaena sanderiana, Phy¬ topathology, 50:648.

Orie, N. G. M., G. S. DeVries, and A. Kikstra, 1960, Growth of Aspergillus in the Human Lung, Am. Rev. Resp. Diseases, 82:6 49.

Pady, S. M., C. L. Kramer, and B. J. Wiley, 1962, Kansas Aeromycology XII: Materials, Methods, and General Results of Diurnal Studies 1959-1960, Mycologia, 51+: 168.

Pratt, H. N., 1938, Seasonal Aspects of Asthma and Hay Fever in New Eng¬ land with Special Reference to Sensi¬ tivity to Mold Spores, New Engl. J. Med., 219: 782.

_ , 1941, Species Specificity

of Alternaria in Asthma and Hay Fever, J. Allergy, 12:431.

Randolph, T. G., and T. L. Squier, 1942, The Incidence of Atmospheric Mold Spores in Relation to Climatic Conditions in Milwaukee, 1935-1941, Wisconsin M. J., 1+1 :987.

Ray, W. W., 1946, Cottonball rots in Oklahoma, Oklahoma Agr. Exp. Sta. Bull. B-300.

Sartory, A., and R. Sartory, 1945, Un cas d’onychomycose du a 1’ Aspergillus fumigatus Fresenius, Bull. Acad, de Med. (Paris), 129:482.

Simon, F., 1938, Allergic Conjunctivitis Due to Fungi, J.A.M.A., 110: 440.

Taub, S. J., 1932, Asthma Due to Yeast (By Ingestion), J. Allergy, 2:586.

Underwood, G. R., 1935, The Impor¬ tance of Fungus as a Cause of Asthma in Nebraska, Nebr. State M. J., 20:400.

Venkatarayan, S. V., and M. H. Delvi, 1951, Black Mould of Onions in Storage Caused by Aspergillus niger, Current Sci., 20: 243.

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Manuscript received January 20, 1971.

INHIBITION OF LEUKOCYTE INDUCED GERMINATION AND TOXIN RELEASE FROM CLOSTRIDIUM BOTULINUM TYPE A SPORES BY CHLOROCRESOL.

J. B. SUZUKI, A. BENEDIK, AND N. GRECZ.

Biophysics Laboratory, Department of Biology , Illinois Institute of Technology,

Chicago, Illinois 60616

Abstract. — Chlorocresol (0.44 mM) inhibits initiation of germination and sub¬ sequent toxin release from spores of Clos¬ tridium botulinum type A Strain 33A in an in vitro guinea pig-polymorphonuclear leukocyte culture as well as in trypticase- peptone broth. In both systems without chlorocresol, labeled C. botulinum spores released previously bound 45Ca at 6-8 hrs indicating that normal germination had occurred. Release of spore-bound botuli¬ num toxin correlated well with spore ger¬ mination, however, toxin release was to¬ tally inhibited for at least 48 hrs by 0.44 mM chlorocresol. These experiments sup¬ port the idea that germination of spores of C. botulinum is an essential step before spores can release spore-bound toxin, thus, becoming pathogenic and that inhibition of germination may be useful in prevent¬ ing this type of pathogenesis.

The in vivo fate of C. botulinum spores upon intramuscular (Keppie, 1951) and intraperitoneal (i.p.) (Booth, et al. 1970, Grecz and Lin, 1968, Grecz et al. 1967) injections have been studied with respect to pathogenicity of these spores. The results to date indicate that in order to release spore-bound toxin with fatal consequences the spores of C. botulinum type A must first germin¬ ate. The changes which accompany germination and outgrowth includes - among others - appearance of stainability, release of calcium, and loss of heat resistance (Murrell, 1961, Hansen, et al. 1970); studies of these changes in relation to patho¬ genesis of C. botulinum spores are summarized below.

Staining of peritoneal exudates from guinea pigs injected i.p. with 10 9 C. botulinum spores revealed macrophages and polymorphonu¬ clear (PMN) leukocytes with pha-

gocytized spores in various stages of germination (Booth, et al. 1971). Further studies on isolated guinea pig PMN leukocyte-C. botulinum spore systems utilizing 45Ca release from labeled spores as indication of germination revealed that spore ger¬ mination occurred at 6-8 hrs (Su¬ zuki, et al. 1971a). Heat resistant (80C, 15 min) spores were converted to heat-sensitive cells at 4-8 hrs in an in vitro leukocyte C. botulinum spore system (Suzuki, et al. 1970), i.e. at a time correlating with 45Ca release. In the same report, using mice i.p. assay, botulinal toxin was released into the media also at 4-8 hrs.

Thus, germination appears to be an essential step for pathogenesis of spores of C. botulinum. There¬ fore, it was of practical and theo¬ retical importance to examine ger¬ mination inhibitors for control of this process. Finding an inhibitor (chlorocresol) which does not affect PMN leukocyte viability and func¬ tion provided us with an important tool to study the mechanism of pathogenesis by C. botulinum spores as described in the present paper.

Materials and Methods

C. botulinum spores:

A culture of C. botulinum type A strain 33A was aseptically inocu¬ lated into a medium containing 5% Trypticase (BBL), 0.5% peptone (Difco), 0.1% sodium thioglycolate (V/V) (TP broth) and 20 micro¬ curies of 45Ca (Tracer Lab. Wal¬ tham, Mass.). 45Ca labeled spores were harvested in a Sorval continu-

358

Suzuki , Benedik and Grecz — Inhibition of C. botulinum

359

ous flow centrifuge (4500 X g, 45 min.) and cleaned with trypsin and lysozyme by the method of Grecz, et al. (1962).

Experimental Animals:

Hartley strain male guinea pigs were raised for 6 generations in a thermostatically controlled room at the Illinois Institute of Technology. They were contained in metal cages with a solid floor and were watered and fed ad libitum with Rockland guinea pig diet with weekly supple¬ ments of lettuce and greens. Guinea pigs attained a weight of 600 + grams before used.

White Swiss mice were raised for 10 generations under similar con¬ ditions and were watered and fed ad libitum with Rockland Rat and Mouse diet. All mice weighed 25 grams before i.p. botulinal toxin assays were performed.

PMN Leukocytes:

A suspension of 85-90% polymor¬ phonuclear (PMN) leukocytes were obtained from guinea pig peritoneal exudates by a modified method of Sbarra and Karnovsky (1959) and explained in detail in these Com¬ munications (Suzuki, et al. 1970).

In vitro System:

One mi of 1 x 108 PMN leuko¬ cytes/ml was added to a siliconized screw-cap tube containing 4 ml Krebs-Ringer Phosphate Medium, pH 7.4 (KRPM) and 3 ml non-im- mune guinea pig serum (Animal Blood Center, Syracuse, N.Y.) and allowed to equilibrate 15 min at 37C in a slowly shaking water bath. A similar system has been described for phagocytic index determinations (Suzuki, et al. 1971). One ml of 1 x 109 45Ca labeled C. botulinum type A spores and 1 ml 4.4 mM chloro- cresol (p-chloro-m-cresol, Fisher) were then added to the guinea pig PMN leukocyte suspension and zero time marked. Chlorocresol is an es¬ tablished inhibitor of spore germi¬ nation (Sierra, 1970, Parker and

Bradley, 1968). At time intervals (0,2,4,6,12,24,36, and 48 hrs), a 1.1 ml sample was pipetted and filtered through a Millipore Swinney (Mil- lipore Filter Corp., Bedford, Mass.) containing a 0.22 micron filter. One- tenth ml of the filtrate was placed in a glass scintillation vial (Amer- sham Seale, Des Plaines, Ill.), 10 ml Bray’s solution (Bray, 1960) added, and kept at 14C until radioactivity was counted. The remaining filtrate (1 ml) was assayed i.p. into mice for presence of botulinal toxin. Con¬ trol mice were passively immunized with botulinal type A antitoxin. An identical in vitro system in TP broth without PMN leukocytes and bot¬ ulinal toxin assay were carried out concurrently and 0.22 micron fil¬ trates treated as above. Radioactivity :

Scintillation vials were counted at 14C for 10 min in a Beckman LS-200 liquid scintillation Counter using a P-32 window.

Results

Figure 1 illustrates 45Ca release appearing in 0.22 micron filtrates

aAverage of 5 experiments.

Background has been subtracted.

Figure 1. Effect of Chlorocresol on Released 45Ca in Filtrate (0.22 u). From Incubation of 1 X 108/ml labeled C. botu¬ linum type A spores and 1 X 1 07/ ml PMN Leukocytes at 37C.a

360

Transactions Illinois Academy of Science

when 1 x 10 9 labeled C. botulinum type A spores are incubated with 1 x 108 guinea pig PMN leukocytes or TP broth both in the presence and absence of 0.44 mM chlorocre- sol. Labeled C. botulinum type A spores in TP broth or in an in vitro PMN leukocyte culture exhibited normal germination patterns sub¬ stantiating a recent report (Suzuki, et al. 1971). Chlorocresol, final con¬ centration of 0.44 mM, introduced into either of the above systems, inhibits germination of C. botulinum spores.

Release of botulinal type A toxin into cell free extracts as determined by mice i.p. assay in presence of 0.44 mM chlorocresol and in con¬ trol systems are summarized in Ta¬ ble 1. From 0-4 hrs of spore incu¬ bation in all conditions, lethal amounts of botulinal toxin could not be detected. At 6-48 hrs with¬ out chlorocresol added to TP broth or leukocyte cultures, C. botulinum spores did invariably release lethal amount of botulinal toxin. How¬ ever, chlorocresol in either system demonstrated a remarkable inhibi¬ tion of toxin release into cell free 0.22 micron filtrates.

Trypan blue dye exclusion tests on PMN leukocytes bathed in 0.44 mM chlorocresol demonstrated neg¬ ligible loss in cell viability from control PMN leukocytes. Phago¬

cytic indices were determined using radioisotope labeling by the method of Suzuki, et al. (1971b). Values for chlorocresol-treated PMN leuko¬ cytes were within normal limits, i.e. 61-64% spores being engulfed.

Discussion

The effect of chlorocresol on bac¬ terial spore germination has been documented (Sierra, 1970; Parker and Bradley, 1968) and is substan¬ tiated in the present communica¬ tion. However, the use of chloro¬ cresol in reducing or completely in¬ hibiting leukocyte induced bacterial spore germination is novel. Further¬ more, in establishing blockage of C. botulinum type A spore germina¬ tion, the requirement for the patho¬ genesis of these spores to first ger¬ minate and then release toxin can be supported.

Using the release of 45Ca from labeled C. botulinum type A spores as evidence of germination, it was observed that germination did in¬ deed occur between 6-8 hrs in TP broth or in vitro in guinea pig PMN leukocyte cultures. Botulinal toxin was found to be released from both systems, also at 6-8 hrs, suggesting either: 1) toxin is released syn¬ chronously from spores as they ger¬ minate, or 2) toxin is released from germinated spores and vegetative

Table 1. Death of i.p. injected mice demonstrating effect of chlorocresol on Release of Type A botulinal toxin in 0.22 Micron Filtrates from C. botulinum type A spores when incubated under the indicated conditions.

Time of incubation

Number of mice expired of a total of 5 injected with cell free filtrates

at 37C

Without

Chlorocresol

With 0.44 m

VI Chlorocresol

Hrs a) b)

109 spores in

109 spores -j-

199 spores in

109 spores -f-

TP-brothc)

108 leukocytes

TP-brothc)

108 leukocytes

0-4

0

0

0

0

6-48

5

5

0

0

a/5 mice injected i.p. with 0.22 micron cell free filtrates taken at each of the following time intervals: 0,2,4,6,12,24,36 and 48 hrs.

t>/All mice protected with anti-botulinal type A toxin survived

c/TP =5% Trypticase, 0.5% peptone, 0.1% sodium thioglycollate broth

Suzuki, Benedik and Grecz — Inhibition of C. botulinum

361

cells after being degraded by leuko¬ cytes or autolysis in TP broth.

The first suggestion conflicts with reports of Tang and Grecz (1968) that toxin was transferred from heat-resistant spores to heat-sensi¬ tive cells and not released into the surrounding medium (phosphate buffer) during this conversion. Therefore, the second suggestion seems more tenable at this point. Preliminary experiments by our lab¬ oratory, utilizing PMN leukocyte metabolic inhibitors further sup¬ port this view (Suzuki, et al. 1971). Relationships of leukocyte metabo¬ lism to Clostridium botulinum patho¬ genesis. (Manuscript in prepara¬ tion) .

Lethal amounts of botulinal toxin were released into the surrounding media at times (6-8 hrs.) correlating with or subsequent to C. botulinum spore germination. Chlorocresol pre¬ vented the release of lethal amounts of botulinal toxin while inhibiting spore germination. Thus, the data strongly suggests that C. botulinum type A spores must germinate be¬ fore becoming pathogenic, and that inhibition of germination may be useful in preventing this type of pathogenesis.

Acknowledgement

This research was supported in part by PHS Grant FD-00358.

References

Booth, R., J. B. Suzuki, and N. Grecz, 1971. Pathogenesis of Clostridium botu¬ linum: In vivo fate of C. botulinum spores. Trans. Ill. St. Acad. Sci. 64: (June).

_ , 1971. Sequential Use of

Wright’s and Ziehl-Neelsen’s Stains for Demonstrating Phagocytosis of Bac¬ terial Spores. Stain. Technol. J±6: 23-26. Bray, G. A., 1960. A simple efficient liq¬ uid scintillator for counting aqueous solu¬ tions in a liquid scintillation counter. Anal. Biochem. 1 :279-285.

Grecz, N. A. Anellis, and M. P. Schneider, 1962. Procedure for clean¬

ing of Clostridium botulinum spores. J. Bacteriol. 84: 552-558.

_ , C. A. Lin, T. Tang, W. L.

So, and L. R. Sehgal, 1967. The na¬ ture of heat-resistant toxin in spores of Clostridium botulinum. Japan. J. Mi¬ crobiol. 11:384-394.

_ , and C. A. Lin, 1967. Prop¬ erties of heat-resistant toxin in spores of Clostridium botulinum 33 A. In M. Ingram and T. A. Roberts (eds) “Botu¬ lism, 1966” Moscow, July 20-22, 1966. Chapman and Hall Ltd. London, pp. 302-322.

Hansen, J. N., G. Spiegelman, and H. 0. Halvorson, 1970. Bacterial Spore Outgrowth: Its Regulation. 1970. Science 168: 1291-1298.

Keppie, J., 1951. The Pathogenicity of the Spores of Clostridium botulinum. J. Hygiene 49:36-45.

Murrell, W. G., 1961. Spore Formation and Germination as a Microbial Re¬ action to the Environment. Sympos. of the Soc. for Gen. Microbiol. XI:100~ 150.

Parker, M. S. and T. J. Bradley, 1968. A reversible inhibition of the germina¬ tion of bacterial spores. Can. J. Micro¬ biol. 14:745-746.

Sbarra, A. J., and M. L. Karnovsky, 1959. The biochemical basis of phago¬ cytosis. I. Metabolic changes during the ingestion of particles by polymor¬ phonuclear leucocytes. J. Biol. Chem. £34:1355-1362.

Sierra, G. 1970. Inhibition of the amino acid induced initiation of germination of bacterial spores by chlorocresol. Can. J. Microbiol. 16: 51-52.

Suzuki, J. B., R. Booth, and N. Grecz, 1970. Pathogenecis of Clostridium botu¬ linum type A: Release of toxin from C. botulinum spores in vitro by leukocytes. Res. Commun. Chem. Pathol. & Phar¬ macol. 1 :691-711.

_ , 1971a. Release of 45Ca

from Clostridium botulinum type A spores in vitro and in vivo as further evidence for germination. Res. Com¬ mun. Chem. Pathol. & Pharmacol. £:16-23.

_ , 1971. Evaluation of Phago¬ cytic Activity by Ingestion of Labeled Spores. J. Infect. Dis. 123:93-96.

Tang, T., and N. Grecz, 1968. Study of Spore Toxin in Germinating Spores of Clostridium botulinum 33A. Bacteriol. Proc. All, p. 2.

Manuscript received April 26, 1971.

ECOLOGICAL STUDY OF A HILLSIDE MARSH IN EAST-CENTRAL ILLINOIS

HAMPTON M. PARKER AND JOHN E. EBINGER

Department of Botany and Bacteriology ,

University of Arkansas, Fayetteville, Arkansas,

and

Department of Botany, Eastern Illinois University, Charleston, Illinois

Abstract. — The marshes studied are located about 5 miles east of Charleston, Coles County, Illinois. They are divided into five areas based on the topography, shading, moisture, and grazing and form habitats in which many plants occur that are rare to east-central Illinois. Nineteen species of woody plants occur in the marshes while the herbaceous vegetation consists of 70 species, of which 3 are fern or fern-allies, 27 are monocots, and 40 are dicots. Two distinct herbaceous com¬ munities exist in the marsh areas depend¬ ing upon the amount of shading. The first is an Impatiens biflora dominated com¬ munity in shaded areas, while one of more open areas consists of Glyceria stri¬ ata, Pilea fontana, Agrostis alba and Ca- rex lurida.

Five miles to the east of Charles¬ ton, Coles County, Illinois, exist a series of small marshes which are unique in that they occur on the side of a hill. These hillside marshes are the result of seepage from a 2 m layer of gravel between two layers of clay. The upper clay layer is about 7 cm thick and allows some water penetration through it, while the lower layer is much thicker and does not allow water to seep through. The water accumulates in the band of gravel and is carried to the outlet areas which form the seepline of the marshes. The result¬ ing marshes form habitats in which many plants occur that are rare to east-central Illinois.

This unique area was first studied by Phipps and Speer (1958), who revealed a number of new plant records for Coles County, Illinois. Parker, Rayhill and Ebinger (1970) also studied this marsh complex and reported 56 new plant records.

The presence of this type of flora indicated that a more intensive study of the area was necessary to determine the relationships between these plants and the habitat. In ad¬ dition, a thorough study was war¬ ranted due to the possible destruc¬ tion of the marshes by the proposed Lincoln Reservoir. When the Reser¬ voir is completed, the normal pool level will be at an elevation of 629 feet above sea level, 30 feet above the level of the marshes.

Method of Study

This study was conducted during the growing seasons of 1967 and 1968. At the beginning of the first growing season 20, one meter rec¬ tangular plots were randomly lo¬ cated throughout the marshes. Every two weeks during the grow¬ ing season the species found in these plots were identified and counted. From this data the density per square meter was determined for each species.

An abundance study of the her¬ baceous plants was undertaken dur¬ ing the growing season of 1968 (Acocks, 1953). A mean distance was determined after a series of measurements were made between individuals of the same species and its abundance class determined by using the following scale:

Symbol Meaning

Distance

VA

Very Abundant

3 inches apart

A

Abundant

6 inches apart

VC

Very Common

1 foot apart

C

Common

1.5 feet apart

VF

Very Frequent

2 feet apart

F

Frequent

3 feet apart

362

Parker and Ebinger — Hillside Marsh

363

F — 6 feet apart

FF Fairly Frequent 12 feet apart

FO Fairly

Occasional 30 feet apart

0 Occasional 50 feet apart

The letter “L” is used after the above letters to indicate local abundance.

A diameter class study of the woody species found in the marsh areas was undertaken during the summer of 1968. These diameter classes were based on diameter at breast height (d.b.h.) and consisted of seedlings (less than 1 inch d.b.h.), saplings (1-4 inches d.b.h.) and trees (more than 4 inches d.b.h.).

The taxonomic nomenclature used throughout this paper follows that of Jones (1963).

Results and Discussion

The marshes are located in a small valley which enters the Em¬ barrass River a short distance south of where Illinois Route 16 crosses the Embarrass River (NE1/4, SW1/4, Section 4, T12N, R10E). The valley floor is at an elevation of 590 feet and the ridges average 655 feet above sea level. The hill¬ sides are relatively steep, with the slope ranging from 40 to 60 degrees. All of the marshes considered in this study are found on the north¬ west-facing hillside and slope to the west from a seepline that is about 10 feet above the valley floor. The soil of the marshes is mainly peat, which in some places reaches a depth of 1-1/4 m.

The vegetation immediately sur¬ rounding the marshes was studied to determine the ecological condi¬ tions and cover types in which the marshes are located (Figure 1). From the lower edge of areas A, B, and C to the creek is a grazed, lowland woods which has an overall varia¬ tion in topography of about 2 m. Juglans nigra, Quercus macrocarpa, and Cary a cordiformis are the domi¬ nant overstory trees here while Cra¬

taegus mollis and C. crusgalli domi¬ nate the understory. From the low¬ er edge of areas D and E to the creek is an ungrazed, lowland woods which has an overall variation in topography of about 2 m and is relatively wet. The dense overstory consists of Juglans nigra, Platanus occidentalis, and Acer saccharum. The understory is dominated by Carpinus caroliniana, Cercis cana¬ densis, and Ostrya virginiana. From the upper edge of areas A, B, and C to the summit of the hill is a heavily grazed, open woods. This area is relatively dry and is domi¬ nated by Quercus alba, Q. velutina, and Q. rubra. The thorny species ( Crataegus spp. and Malus ioensis ) dominate the understory. From the upper edge of areas D and E to the summit of the hill is an ungrazed, mesic woods. The vegetation here is more mature than in any other region surrounding the marshes. The dominant species of this hill¬ side are Quercus alba, Q. velutina, Q. rubra, and Acer saccharum while the understory is dominated by Ostrya virginiana, Cornus florida, and A. saccharum.

A total of 19 species of woody plants are found in the marshes, but none has a d.b.h. above 4 inches. Table 1 lists the actual num¬ ber of seedlings and saplings found in each of the marsh areas studied. The herbaceous vegetation is also not extremely diverse and only 70 species are present. Of these, 3 are fern or fern-allies, 27 are monocots and 40 are dicots. The commonly encountered herbaceous species are listed in Tables 2 and 3.

Two major herbaceous communi¬ ties exist in the marshes. These re¬ sult from the amount of shading due to the overstory trees and vary to some extent in composition de¬ pending upon local habitat condi¬ tions. In the parts of the marshes

364

Transactions Illinois Academy of Science

Figure 1. Map showing the marsh areas studied

Table 1. Total number of seedlings and saplings of the various species of woody plants found in the zones of the marsh areas studied.

Parker and Ebinger — Hillside Marsh

365

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Transactions Illinois Academy of Science

Table 2. Abundance classes and density per square meter for the dominant species of the densely shaded areas of the hillside marshes. For explanation of abundance class symbols see the Method of btudy section of this paper.

Species

Abundance Classes by Areas

Density

per

square

meter

Area A1

Area B1

Area Cl

Area E

Impatiens biflora

VA

VA

A

A

17.0

Glyceria striata

C

VC

F-

F

4.8

Carex lurida

FF

VF

VC

CL

0.4

Cardamine bulbosa

FF

C

FF

VF

2.0

Pilea fontana

• • •

AL

VA

FF

0.2

Caltha palustris

F

F

...

F

2.0

Solidago patula

F-

FF

FF

F

4.0

Aster laterifiorus

FF

F

F-

FF

1.6

Oxypolis rigidior

0

. . •

F-

FF

1.0

Equisetum arvense

VF

FF

...

0

0.2

Cinna arundinacea

0

FF

F-

FF

Others

• • •

. . .

. . .

. . .

4.4

TOTAL

. . .

• . •

• • •

. . .

37.6

that are densely shaded, Impatiens biflora is the dominant species en¬ countered (Table 2). In all of the shaded zones of areas A, B, C and E this species varies in abundance classes from abundant to very abun¬ dant. Numerous seedlings of this species occur in the shaded parts of the marsh early in the growing sea¬ son, but most do not reach matur¬ ity. Impatiens biflora averages 17 individuals per sq m which accounts for nearly half of the density ob¬ served in the plots of the shaded zones. The commonly encountered associated species are Glyceria stri¬ ata , Carex lurida, Cardamine bul- bosa, and Pilea fontana. In most in¬ stances these species are scattered, rarely exceeding an abundance class of frequent. The only exception be¬ ing Pilea fontana which replaces Impatiens biflora in a few small areas.

At the edge of the shaded parts of the marshes a very narrow tran¬ sition zone exists where Impatiens biflora rapidly decreases in abun¬ dance while Glyceria striata and Ca¬ rex lurida greatly increase, becom¬

ing very frequent to abundant. Al¬ so, Agrostis alba and J uncus dud- leyi, which did not occur in the shaded parts of the marsh, become common to very abundant. Pilea fontana also becomes more abun¬ dant, but always as small plants, less than 3 inches tall, that persist under the larger herbaceous plants. This second community is consid¬ ered to be of an open overstory type and is seldom shaded through the day (Table 3). It is dominated by Glyceria striata , Pilea fontana, Agrostis alba and Carex lurida which collectively make up about one-half of the total density per sq m and are classed as very abundant to very frequent.

The hillside marshes are divided into five areas based on the topog¬ raphy, shading, moisture, and graz¬ ing. Starting with the southernmost marsh, the various marsh areas are listed below (Figure 1).

Area A. — Ungrazed, Spot Marshes: This group of three marshes was fenced in 1956 and has not been grazed since that time. Two dis¬ tinct zones exist here, depending

Parker and Ebinger — Hillside Marsh

367

Table 3. Abundance classes and density per square meter for the dominant species of the open areas of the hillside marshes. For explanation of abundance class symbols see the Method of Study section of this paper.

Species

Abundance Classes by Area

Density

per

square

meter

Area A2

Area B2

Area C2

Area C3

Area C4

Glyceria striata

VC

VC

A

A

A

16.5

Pilea fontana

VA

VA

VA

VCL

32.3

Agrostis alba

VA

C

C

VC

C

9.6

Carex lurida

VFL

A

VC

A

A

7.1

Juncus dudleyi

....

A

VC

C

VC

5.1

Equisetum arvense

• • • •

....

C

VC

VC

5.3

Selaginella apoda

....

VC

C

FF

FF

4.5

Eleocharis erythropoda

....

A

A

. , . .

....

13.6

Eupatorium perfoliatum

F-

F

FF

F

F

3.6

Solidago patula

F

F

F

VF

VF

2.9

Pedicularis lanceolata

VF

F-

FF

F

VF

1.1

Caltha palustris

0

VC

FF

A

2.4

Aster lateriflorus

F

F-

FF

....

FF

2.0

Chelone glabra

AL

0

F-

F

F-

1.3

Lobelia siphilitica

F-

F-

FF

F-

FF

1.2

Scirpus atrovirens

C

VC

VFL

VFL

VFL

.6

Oxypolis rigidior

VF

F-

F-

F-

F

.7

Carex vulpinoidea

FL

F

CL

FL

CL

.5

Others

....

....

• • • •

....

....

26.5

TOTAL

....

....

....

....

....

136.8

on the degree of shading due to trees surrounding the marshes. The shaded zone is dominated by the Impatiens biflora community while Salix discolor is the dominant woody species (Table 1). The presence of woody species is probably due to the lack of grazing in this zone for the last 14 years. A small section of the southern-most spot marsh is in full sunlight (Table 3, Area A2). The herbaceous vegetation of this zone is dominated by Glyceria stri¬ ata, Agrostis alba, and a third spe¬ cies of grass, Leersia oryzoides which did not occur in the other open marsh areas.

AreaB. — Grazed Marsh: Thirty me¬ ters to the north of Area A is a boot-shaped marsh consisting of 0.17 acres which has been grazed for at least the past 20 years. This marsh is extremely wet and stand¬ ing water is found in the depres¬ sions most of the year. Its northern

and extreme eastern edge are heav¬ ily shaded by surrounding trees. This shaded zone is dominated by Impatiens biflora while Pilea f on- tana and Glyceria striata are impor¬ tant associated species (Table 2, Area Bl). The remainder of this marsh area is in full sunlight for most of the day and is dominated by the Glyceria striata, Pilea f on- tana, Agrostis alba and Carex lurida community (Table 3, Area B2). Al¬ so, Scirpus validus is very abundant here but did not occur in the other marsh areas. All of the woody plants observed in the grazed marsh oc¬ curred here (Table 1).

AreaC. — Diverse Marsh: Three me¬ ters to the north of Area B is the marsh studied by Phipps and Speer (1958). It consists of 0.20 acres and has not been grazed since March of 1967, when it was fenced. The topography varies from a flat area to a hillside with a slope of about

368

Transactions Illinois Academy of Science

45 degrees and has an overall fall of about 4 m from the seepline to the lowest point.

The typical shaded zone vegeta¬ tion, dominated by Impatiens bi¬ flora, exists only in the extreme southern corner of this marsh (Ta¬ ble 2, Area Cl) while the remainder of this marsh is dominated by the Glyceria striata, Pilea fontana, Ag- rostis alba, Carex lurida community (Table 3, Area C2, C3 and C4). Three distinct zones exist in the open parts of this diverse marsh de¬ pending upon topography and mois¬ ture and, as a result, minor varia¬ tions in this dominant plant com¬ munity occur. These are a Low, Open Zone, a Hillside, Open Zone and a High, Open Zone (Figure 1).

The Low, Open Zone is about 22 m long, averages 3 m wide, and oc¬ curs in a depression at the base of the hill in which water from the hillside seep accumulates. The wa¬ ter in the depression is about 2 dm deep and remains as a pool for the entire year. Besides the species typi¬ cally found in the open area com¬ munity, Eleocharis erythropoda, Bi- dens cernua, Carex festucacea, and Sagittaria latifolia are abundant to very common in this zone.

The Hillside, Open Zone extends from the seepline to the Low, Open Zone and has a fall of about 4 m from the seepline to the base of the hillside with a slope of about 45 degrees. It is quite wet but without standing water, is very soft and muddy near the seepline, and is in full sunlight for most of the day. More woody individuals and spe¬ cies occur here than in any other part of the diverse marsh because grazing was impossible owing to the steep slope (Table 1).

The High, Open Zone of the di¬ verse marsh area lies between the seepline and the upper edge of the Hillside, Open Zone, is in full sun¬ light for most of the day and has

an overall fall of about 1.5 m. The area is 20 m long, 7 m wide, has a northwest exposure, and is very wet with standing water in depres¬ sions throughout the year. Besides the typical open areas species, Carex vulpinoidea and C. stipata are com¬ mon while Caltha palustris is abun¬ dant throughout this zone. Few woody plants exist here (Table 1), although Salix discolor is represent¬ ed by a few individuals near the fence that separate this zone from Area D.

AreaD. — Wooded, Ungrazed Marsh: This marsh is an extension of the High, Open Zone of Area C. It con¬ sists of 0.33 acres and averages 45 m long and 20 m wide. It has about a 5-degree slope to the northwest and an overall fall of about 2 m from the seepline to an outlet de¬ pression that drains into the creek. Phipps and Speer (1958) reported that this area had not been grazed in 1957, and it appears that no grazing has taken place since that time.

Saplings of Salix discolor are very common on the slope above the outlet depression, while Salix rigida with 8,884 seedlings dominate this depression at the base of the slope. Carpinus caroliniana and Ulmus americana are commonly found near the seepline where the willows are fairly sparse and some shading oc¬ curs. Rhus vernix exists as a single clump on the slope associated with Salix discolor. A few specimens of Sambucus canadensis and Ribes americanum are found in the outlet depression at the base of the marsh (Table 1).

The herbaceous vegetation of this area is extremely diverse, and be¬ cause of the degree of shading, ap¬ pears to be intermediate between the two common communities oc¬ curring in the rest of the marsh areas. Also, some species are found here and in no other part of the

Parker and Ebinger — Hillside Marsh

369

marshes. These include Iris shrevei, Carex hyalinolepis , Apios ameri- cana and Lysimachia ciliata.

Area E. — Ungrazed, Shaded Spot Marshes: Northeast of Area D are three small spot marshes which are similar in topography to the spot marshes of Area A. These marshes, which average 100 sq m in size, slope to the northwest with about a 5-degree incline. They are ex¬ tremely wet with standing water in the depressions throughout the year. The herbaceous vegetation of this area is very similar to that of the shaded zone of Area A (Table 2).

Conclusions

As a result of variation in topog- graphy, available moisture, light penetration, and grazing, certain trends can be observed in the marsh areas studied. Many indications suggest that grazing has been very effective in controlling the invasion of woody plants into the marshes. This can be determined by com¬ paring the various marsh areas which have been grazed to varying extents during the past 14 years. The Spot Marshes (Area A) and the Wooded (Area D) have not been grazed for at least 14 years (Phipps and Speer, 1958), and in both areas numerous woody plants occur. In contrast, the Grazed Marsh (Area B) and the Diverse Marsh (Area C) have been subjected to extensive grazing for at least the past 20 years. In both, few woody plants exist, except in areas that are not accessible to cattle.

Distinct herbaceous communities are present in the marsh areas stud¬ ied. The shaded zones of Areas A, B, C, and E are similar in ecological

condition and associated plant spe¬ cies. In these zones, which are shaded for most of the day, Im- patiens biflora dominates and is classed as abundant to very abun¬ dant. Few other species occur in this wet, shaded situation, and all are in low abundance classes.

The open zones of the marsh areas studied are similar ecologi¬ cally and the associated species are similar though they vary in abun¬ dance. The dominant species here are Glyceria striata , Pilea fontana, Agrostis alba and Carex lurida. The open nature and the similarity of plants that occur here tend to place these zones in the same habitat type.

If part of the area continues to be protected from grazing and if the seep continues to produce an abundant amount of water, it is very possible that this unique com¬ munity will persist. Under these conditions most of the plants will survive and continue to flourish. One exception may be Rhus vernix, which is now limited to one clump and does not appear to be repro¬ ducing.

Literature Cited

Acocks, J. 1953. Veld types of South Af¬ rica. Bot. Survey Mem. No. 28, Dept. Agric., Div. Bot. Pretoria, Union of South Africa.

Jones, G. N. 1963. Flora of Illinois. 3rd ed. Amer. Midi. Nat. Monog. 7:vi + 401 pp.

Parker, H. M., S. M. Rayhill, and J. E. ebinger. 1970. Additions to the flora of Coles County, Illinois. Trans. Ill. St. Acad. Sci. 63:375-378.

Phipps, R., and J. Speer. 1958. A hillside marsh in east-central Illinois, Trans. Ill. St. Acad. Sci. 51:37-42.

Manuscript received May 4, 1971.

THE EFFECT OF DIMETHYL SULFOXIDE ON HUMAN ERYTHROCYTE MEMBRANE

DENNIS M. KALLVY AND JOSEPH C. TSANG

Department of Chemistry, Illinois State University, Normal, Illinois 61761

Abstract. — Isolated human erythrocyte membranes were submitted to sonication in the presence and absence of dimethyl sul- f oxide. Solubilized protein fractions con¬ taining high sialic acid and low lipid con¬ tent were obtained. The possible role of DMSO as a cryoprotective agent in a mem¬ brane system was discussed.

The human erythrocyte mem¬ brane has been the subject of inten¬ sive research because of its ready availability and ease of isolation. Knowledge of the precise arrange¬ ment of its components may open the way for treatment of blood di¬ seases such as hereditary elliptocy- tosis and hereditary spherocytosis, which are suspected to be caused by membrane defects. The struc¬ ture of erythrocyte membranes may also be common to that of other bi¬ ological membranes. It is therefore hoped that other diseases may be explainable in terms of membrane activity.

Adequate methods of solubiliza¬ tion of the membrane components must be developed before the struc¬ ture of the membrane can be eluci¬ dated. Several attempts to develop suitable procedures have been made (Maddy, 1970). These include or¬ ganic solvent extractions (butanol, 2-chloroethanol, phenol, aqueous pyridine and pentanol), detergent solubilization (cholate, deoxycho- late, sodium dodecyl sulfate and Triton X100), as well as hydrogen bond-breaking reagents (urea and guanidine hydrochloride). However, these reagents either lack specificity in solubilizing a particular compo¬ nent, forming complexes with the membrane components, or dena¬ ture proteins. Therefore, it is im¬ portant to explore the potential of other reagents for the specific re¬

moval of membrane components. For example, dimethyl sulfoxide (DMSO) has been known for its use¬ fulness in the solubilization of gly¬ cogen (Whistler and DeMiller, 1962) and selective extraction of lipopoly- saccharide from the outer mem¬ brane of Gram-negative bacteria (Adams, 1967).

It would be of interest to deter¬ mine the chemical nature of the components extractable from hu¬ man erythrocyte membranes by dimethyl sulfoxide. This report rep¬ resents a study of the extraction procedures and the chemical anal¬ yses of the extracts as well as the residues.

Materials and Methods

Hemoglobin-free human erythro¬ cyte membranes were isolated from outdated blood (Peoria Red Cross, Peoria, Illinois) according to the procedure of Dodge, et al., 1963. The cells were washed three times in isotonic 310 ideal milliosmolar phosphate buffer, pH 7.4, lysed overnight in hypotonic 20 ideal mil¬ liosmolar phosphate buffer and washed with hypotonic buffer until free of hemoglobin.

After dialysis and lyophilization, a 200 mg sample of membrane was added to 50 ml of 0.1 M phosphate buffer, pH 7.4, and stirred to ho- mogeniety for 1-2 hours. An equal volume (50 ml) of DMSO was slow¬ ly added, stirred to homogeniety for 1 hour, and sonicated (Branson Sonicator Model W140D) in 50 ml aliquots at 4° C at 60 watts for 6 minutes. Centrifugation was per¬ formed at 20,000 x g for 20 minutes at 4° C after the sonicates were combined. The supernate was de-

370

Kallvy and Tsang — DMSO on Erythrocytes

371

canted, and the precipitate was re¬ constituted for 2-3 hours in 25 ml of 0.1 M phosphate buffer, pH 7.4. An additional portion of DMSO (25 ml) was slowly added to the suspension; sonication and centrifu- tion were executed as before. The two combined supernates (Frac¬ tion S) and the reconstituted preci¬ pitate (Fraction P) were dialyzed against distilled water for one week and lyophilized.

A control sample was treated exactly as described above, except an equal volume of 0.1 M phos¬ phate buffer, pH 7.4, was added to the buffer-membrane suspension in place of the equal volume of DMSO. The supernate and precipitate of this treatment were recovered as NS and NP, respectively.

Protein (Lowry, et al., 1959), hex- osamine (Rondle and Morgan, 1955), hexoses (Koehler, 1952), si¬ alic acid (Warren, 1959), and phos¬ phorous (Bartlett, 1959) were de¬ termined. Total lipids were ex¬ tracted with chloroform methanol (2:1 v/v) in a Soxhlet apparatus and determined gravimetrically.

Results and Discussion

Dimethyl sulfoxide (DMSO), an aprotic solvent, is used in biologi¬ cal systems to protect cells against freezing and radiation damage (Far- rant, 1965; Chang and Simon, 1968). One of the hypotheses to explain the cryoprotective and radio-pro¬ tective action of DMSO assumes that this solvent prevents changes in the cell's lipoprotein membranes and stabilizes lipoprotein complexes (Keysary and Kohn, 1970). Five to ten percent DMSO is widely used as an additive for the protection of animal cells during freezing stor¬ age. However, when continuously present, these concentrations might be toxic. It seems therefore para¬ doxical that a toxic substance should be protective. A similar situation

occurs in the bacterial endotoxin systems, which are toxic over high concentration and protective at low concentrations. Furthermore, it is puzzling that a reagent known to extract carbohydrates (Whistler and DeMiller, 1962; Adams, 1967) would serve as a stabilizing agent for the membrane system which is known to consist of protein, lipids and car¬ bohydrates (Bakerman and Wase- miller, 1967).

In experiments reported here, a high concentration of DMSO (50%) in a buffered solution was used to extract human erythrocyte mem¬ branes. DMSO extraction along with sonic treatment was found to solubilize 28.7% while sonic treat¬ ment alone solubilized 23.6%. In both cases, the ratio of the amount of residue to amount solubilized was approximately 2:1 (Table 1 and 2). Since the presence of DMSO did not affect the relative ratio of resi¬ dues to solubilized material, it ap¬ pears that sonication alone was re¬ sponsible for the extraction effi¬ ciency. In order to determine the chemical nature of the fractions, chemical analyses were performed (Table 1 and 2). In both cases sig¬ nificant amounts of total carbohy¬ drates (15%), especially sialic acids (5%) was found in the solubilized fractions. On the other hand, there was a slightly larger amoant of total free lipids in the residues (60%) than in the solubilized fractions (45%). There was no significant difference in the amount of pro¬ teins (43%) in the residues which have a similar protein composition as the starting intact membranes. The sialic acid-rich fractions (Frac¬ tions S and NS) resemble those pre¬ pared by aqueous pyridine extrac¬ tion (Blumenfeld, 1968) and by pronase treatment (Ohkuma and Furuhata, 1969) of human erythro¬ cyte membranes. From these stud¬ ies, it seems that there are two types

372

Transactions Illinois Academy of Science

Table 1. Chemical Composition of Human Erythrocyte Membrane Fractions After Sonication Treatment in the Presence of Dimethyl Sulfoxide

Fractions

S

P

Intact Membrane

Experimental

Literature

Yield (%)

28.7

49.6

0.7608*

Protein (%)

34.9

42.2

46.1

55.0**

Total Lipid (%)

50.0

61.0

37.2

35.0**

Phosphorous (%)

1.40

1.29

0.90

1.10***

Total Carbohydrate (%)

15.19

11.28

9.49

10.0**

Hexosamine (%)

5.00

3.92

3.00

Sialic Acid (%)

5.35

2.02

2.57

Hexoses (%)

4.84

4.34

3.92

*Number of grams from 600 ml blood **Bakerman, et al., 1967 ***Lauf and Poulik, 1968

Table 2. Chemical Composition of Human Erythrocyte Membrane Fractions

After Sonication

Fractions

NS

NP

Intact Membrane

Experimental

Literature

Yield (%)

23.6

49.8

0.7608*

Protein (%)

33.0

44.1

46.1

55.0**

Total Lipid (%)

45.0

60.0

37.2

35.0**

Phosphorous (%)

1.46

1.08

0.90

1.10***

Total Carbohydrate (%)

15.89

9.32

9.49

10.0**

Hexosamine (%)

4.90

3.82

3.00

Sialic Acid (%)

5.10

1.70

2.57

Hexoses (%)

5.89

3.80

3.92

*Number of grams from 600 ml blood **Bakerman, et al., 1967 ***Lauf and Poulik, 1968

of proteins in the erythrocyte mem¬ brane, one is high in sialic acid and low in lipid; the other low in sialic acid and high in lipid. In both the aqueous pyridine and pronase prep¬ arations, the fractions rich in sialic acids bear antigenic determinants of the MN blood group system. It is not known if the sialic acid-rich fractions isolated by DMSO treat¬ ment would share this immunologi¬ cal property.

It is surprising that DMSO failed to extract much free lipid. It would be expected that sonication alone in an aqueous medium would be less effective and DMSO, being an aprotic organic solvent, more effec¬

tive in removing lipid materials. On the other hand, this unique prop¬ erty of DMSO may explain the pos¬ sible stabilization effect on the lipo¬ protein complexes of the membrane.

It has been reported (Rosenberg and McIntosh, 1968) that sonica¬ tion does not alter the molecular membrane structure. The solubili¬ zation effect by sonication was sug¬ gested to be caused by the disin¬ tegration of the total membrane system and reconstitution to small unit-membrane pieces which did not sediment under high centrifu¬ gation force. These solubilized unit- membranes were shown to have similar chemical composition as the

Kallvy and Tsang — DMSO on Erythrocytes

373

intact membranes. Our results are not consistent with their findings. Similar to the treatment in the presence of DMSO, sonication alone was able to release a sugar-rich moiety (Fraction NS). The exact nature of the solubilized material in the presence and absence of DMSO may not be known until their ho¬ mogeneity is established by column fractionation or polyacrylamide disc electrophoresis. These solubilized fractions may well be a complex (or complexes) of lipids, proteins, gly¬ coproteins and glycolipids. One thing is certain, however, that there is an easily releasable sialic acid-rich moiety in the human erythrocyte membrane.

Although DMSO extraction did not provide a clear-cut selectivity for any particular components of the erythrocyte membrane, the in¬ ability of DMSO to remove lipid to any great extent does not con¬ tradict with the suggestion that membrane lipids may be the site or sites for freezing injury to cellular systems (Livne, 1969) which could be protected by low concentrations of DMSO.

Acknowledgement

The authors thank the Red Cross Blood Center, Peoria, Illinois for the supply of out-dated blood.

Adams, G. A. 1967. Extraction of Lipo- polysaccharide from Gram-negative Bacteria with Dimethyl Sulfoxide. Can. J . Biochem., 55: 422-426.

Bakerman, S., and Wasemiller, G. 1967. Studies on Structural Units of Human Erythrocyte Membrane. I. Separation, Isolation, and Partial Characterization. Biochemistry, ^ :1 1 1 0-1 113. Blumenfeld, O. O. 1968. The Proteins of Erythrocyte Membrane Obtained by Solubilization with Aqueous Pyridine Solution. Biochem. Biophys. Res. Comm., 30: 200-205.

Bartlett, G. B. 1959. Colorimetric As¬ say Methods for Free and Phosphory- lated Glyceric Acid. J. Biol. Chem., 234:469-471.

Chang, C., and Simon, E. 1968. The Ef¬ fect of Dimethyl Sulfoxide (DMSO) on Cellular Systems. Proc. Soc. Exp. Biol. Med., 128:6 0-66.

Dodge, J. T. Mitchell, C., and Hana- han, D. J. 1963. The Preparation and Chemical Characteristics of Hemoglo¬ bin-free Ghosts of Human Erythro¬ cytes. Arch. Biochem. Biophysics, 100: 119-130.

Farrant, J. 1965. Mechanism of Cell Damage during Freezing and Thawing and its Prevention. Nature, 205: 1284- 1287.

Koehler, L. H. 1952. Differentiation of Carbohydrates by Anthrone Reaction of Rate and Color Intensity. Anal. Chem., 25: 1576-1579.

Keysary, A. and Kohn, A. 1970. Effects of Dimethyl Sulfoxide on Macromole- cular Synthesis in Animal Cells in Vitro and their Relevance to Cryoprotec- tion. Chem.-Biol. Interactions, 2:381- 390.

Lauf, P. K., and Poulik, M. D. 1968. Solubilization and Structural Integrity of the Human Red Cell Membrane. Brit.J. Haemat., 15:191-202.

Livne, A. 1969. Membrane Lipids as Site for Freezing Injury. Israel J. Chem., 7: 152p.

Lowry, O. H. Rosebrough, N. J., Farr, A. L., and Randall, R. J. 1951. Pro¬ tein Measurement with the Folin Re¬ agent. J. Biol. Chem., 193: 265-275.

Maddy, A. H. 1970. Erythrocyte Mem¬ brane Proteins. Seminars in Hematol¬ ogy, 7: 275-295.

Ohkuma, S., and Furuhata, T. 1969. Role of Sialic Acid Residues in Some Properties of Sialoglycopeptide Re¬ leased by Pronase from Human Ery¬ throcytes. Proc. Japan Acad. Sci., 55: 417-421.

RondleM. C. M., and Morgan, W. T. J. 1955. The Determination of Glucosa¬ mine and Galactosamine. Biochem. J., 61: 586-589.

Rosenberg, S. A., and McIntosh, J. R. 1968. Erythrocyte Membranes; Effects of Sonication. Biochim. Biophys. Acta., 163: 285-289.

Warren, L. 1959. The Thiobarbituric Acid Assay of Sialic Acids. J. Biol. Chem., 23U1971-1975.

Whistler, R. L., and Demiller, J. N. 1962. Extraction of Glycogen with Di¬ methyl Sulfoxide. Archiv. Biochem. Bio¬ phys., 98: 120-123.

Manuscript received April 11, 1971.

LAKE SHORE EROSION

WYNDHAM J. ROBERTS

State Water Survey, Urbana, Illinois 61801

Abstract. — Erosion of lake shores by boat traffic is a serious problem which needs to be given more attention. This erosion contributes to the filling in of res¬ ervoirs. A gabion wall is recommended.

Present concern with ecological problems tends to obscure natural geological processes which have been proceeding since the beginning of time and which man is accelerating. This is apparent when a lake is con¬ structed, for any impoundment rep¬ resents a block to Nature’s orderly process. The arresting of water flow behind a dam means that the sedi¬ ment load carried by moving water cannot be held and is deposited on the floor of the lake. This process

is largely overlooked because it is unseen and, depending upon the size of the area contributing runoff, not observed until considerable stor¬ age volume has been lost to sedi¬ ment.

Although most of the deposited sediment has been transported from the total watershed, a portion con¬ sists of material eroded from head¬ lands in the lake that have been under attack of wave-generated winds and ice pressure, like the slope shown in Figure 1. When suffi¬ cient sediment is eroded to make a protective beach, the rate of head¬ land erosion is greatly reduced.

Figure 1. Lake headland eroded by wave action.

374

Roberts — Lake Shore Erosion

375

However, the process results in the loss of considerable land, generally high-value promen tary land. Usu¬ ally, land owners try to protect ex¬ posed erodable areas with seawalls, such as the low barrier pictured in Figure 2. Unless they are well con¬ structed, they require almost con¬ stant upkeep at great expense.

Man contributes markedly to this type of erosion through the uncon¬ trolled use of power boats on lakes. This is especially noticeable on nar¬ row lakes where wave-generated waves reach the shore with maxi¬ mum crests and undiminished force. Even with lake police protection, man-made lakes continue to be dam¬ aged by fast boat traffic throughout the months of heavy recreational use. One has only to observe the recreational use of municipally

owned and operated lakes to know that boating enthusiasts frequently overstep the rules of safe operation. Especially is this true for water skiers. In making 180 degree turns it is frequently necessary for the boat to make a circular course that almost intersects the shore. Waves generated by this maneuver cause considerable erosion.

Normal wave action generated by boats cruising 100 yards from shore reaches the shore with a dy¬ namic pressure of less than 5 pounds per square foot. A speeding boat pulling a skier at 20 miles per hour as shown in Figure 3 and at a dis¬ tance of 20 yards from shore can send shoreward waves that exert pressures on the shore of 500 to 1000 pounds per square foot. Ob¬ viously, the answer to this kind of

Figure 2. Low wood barrier wall.

376

Transactions Illinois Academy of Science

erosion is the enforcement of stricter rules for boat operation. Since dy¬ namic pressure varies as the cube of the wave height, erosion can be reduced considerably if waves have more distance to travel before they intersect the exposed shore and break with less force.

In view of the destruction of lake shore lines that could be attribut¬ able to boat traffic, it is strange that there are not uniform codes controlling all lake boat traffic si¬ milar to the motor vehicle laws. The tendency has been for a situation to develop to a critical stage and then investigate the history of older lakes and adopt some of rules for¬ mulated for their protection. Some general practices can be learned in this way.

Rules and Regulations

Article XXIII of the Rules and Regu¬ lations of the Illinois Department of Con¬ servation states that it is unlawful for any person to use an outboard motor on any state-owned lake under its jurisdiction that has less than 60 acres of water. On the larger state-owned lakes, the limita¬ tion applies to use of an outboard motor of a size larger than 10 horsepower. These regulations are established in accordance with Section 4 of the Fish Code of Illinois, and persons violating provisions are sub¬ ject to the penalties provided by the Code. The laws supplementing these rules are to be found in Chapter 56 of the Illi¬ nois Revised Statutes.

Rules governing boat traffic on muni¬ cipally owned lakes vary greatly. Lake Bloomington, a 600-acre lake owned by the City of Bloomington, has a limit of 25 horsepower for motor boat operation. Lake Sara, a 735-acre lake built by the Effingham Water Authority, has an up¬ per limit of 75 horsepower for its boat traffic. Lake Springfield, which covers an area of 5000 acres, has no horsepower

Figure 3. Pleasure boat producing waves.

Roberts — Lake Shore Erosion

377

limit, with several boats having motors in excess of 300 horsepower. However, hydroplanes are not allowed, and no boat may speed in excess of 35 miles per hour.

License Fees

There is a charge for operation of any boat on practically all Illinois lakes. The State Department of Conservation charges fees varying from $12.50 per sea¬ son for power boats to $25.00 for pontoon boats.

Approximately 3100 boats are licensed to operate on Lake Springfield. There are regularly 2200 locally-owned boats with licenses, and the remainder come from great distances, often beyond the state’s borders. Annual fees vary from $2.00 for a rowboat less than 16 feet in length and having no motor to $10.00 for a boat that is powered by a motor in the range of 51 to 74 horsepower.

At Lake Decatur, which has an area of slightly more than 2400 acres, the sched¬ ule of fees blankets all outboard motors under 15 horsepower at $7.50 annually, and $10.00 for larger sizes. Boats with inboard motors are charged $12.50 if the

size is under 175 horsepower and $15.00 for more powerful engines. Sailboats are all charged $7.50.

Such fees are collected by city clerks, and the funds are used to help defray the cost of policing the lakes. Violations are punishable with fines, and these also help toward the same costs.

Erosion Barrier Construction

Generally, shore property along Illi¬ nois lakes is not provided with protective devices until erosion has progressed con¬ siderably. The disturbed owner will then observe the protective devices already installed on other shore properties and will copy them without regard to appli¬ cability to a specific situation. Figure 4 is typical of improper planning to curtail bank erosion.

At Lake Sara, wooden seawalls are popular. They are composed of wood posts 6 feet long driven vertically into the shoreline and protruding above spill¬ way crest elevation one or two feet. Wood planks, each 2 by 6-inches by 12 feet, are nailed horizontally to the posts. The wall is also supported from the shore side with

Figure 4. Inadequate barrier fails to stop bank erosion.

378

Transactions Illinois Academy of Science

coarse filler material such as bricks, rocks and cement blocks. Most owners have had relatively good experience with this type of construction, but any unusual high water tends to lift the structure out of the water as shown in Figure 5.

One erosion barrier that has had a fair measure of success consists of a contin¬ uous concrete wall approximately 6 inches wide and extending from the ground ver¬ tically to a height 6 inches above spillway crest elevation. The wall is strengthened with reinforcing rods and supported on the shore side with rocks, bricks, and broken concrete. Where rods have not been tied together, failure has occurred in the wall at the discontinuity, as shown in Figure 6a. A similar wall may be strengthened at regular intervals with corrugated metal area-way projections 2 feet deep and 3 feet in diameter. They are connected to the main wall with trellis-type wire and filled with concrete. The semicircular shape of the way-walls tends to absorb much of the wave energy. The wall shown in Figure 6b has with¬ stood three years of battering.

Property owners have experienced only

limited success with protective barriers installed along their water lines, and in many cases failure has occurred within a year. Since permanent walls, such as driven sheet piling, are expensive, it is imperative that effective yet less costly devices be investigated.

One form of protection that should prove beneficial is the gabion. This is a steel wire mesh box made of a complete and continuous fabric, as shown in Figure 7, and filled with stones or rocks. It may be constructed in varying sizes and shapes, although in practice the horizontal width should be at least three feet. The height can be fractions of the width and the length up to four multiples of the width. The wire mesh, having a minimum size of U.S. Steel Wire Gage No. 11, should be galvanized with a minimum zinc coat¬ ing of 0.80 oz./per square foot of wire sur¬ face. The area of the mesh openings should not exceed 8 square inches, and the maximum linear dimension of the opening not exceed 4.5 inches. Strength, elasticity, and flexibility are necessary requirements of gabions so they should be divided into compartments with dia-

Figure 5. Wood barrier uprooted by high water.

Roberts — Lake Shore Erosion

379

phragms of the same mesh and gage as the main body. The smaller cells diminish the internal movement of the rock fill. Single unit construction is necessary so that edges of the base, lid, ends, and sides have the same strength and flexi¬ bility as the body of mesh.

Cost of erosion Barriers

There is considerable variation in the prices owners have reported for their bar¬ rier installations. The wooden post vari¬ ety with horizontal boards costs about $40 a lineal foot when installed by a con¬ tractor. Many do-it-yourselfers have built such walls for about $14 a lineal foot.

Contractors have installed gabion walls at a cost of $69 per running foot. Figure 8 shows a typical gabion wall. A sheet steel type of wall driven to a depth of ten feet costs about $1800 per lineal foot. While this is the best protection, especial¬ ly where salt water and wave action is great, the gabion has attributes that make it preferable for preventing shore erosion at small lakes. The rock-filled walls are porous and therefore not as sub¬ ject to hydrostatic pressure deformations, especially concrete ones. As they fill with silt they support plant life and blend in

Figure 6a. Wall failure where reinforcing rods were omitted.

with the natural surroundings. The gabion type of structure protects the headland or “toe” of the shoreline from under¬ mining, which causes most bank failures due to wave action.

Conclusions

Shoreline erosion is a continuing pro¬ cess, even after adequate beaches have developed. However, the average lot own¬ er knows that he will lose some of his water-line property land to wave erosion and generally makes an attempt to pro¬ tect his shoreline. Unfortunately, the pres¬ ent make-shift schemes used by most property owners do not begin to do an adequate job of protecting lake shore property lines, and costs for this work are expanding. There is need for more adequate information on workable shore protective devices. Cost of experimental devices could be met by the state through bills proposed by legislators for experi¬ mental barriers built at specific locations. A state department, such as the Depart¬ ment of Conservation, could then make the results available to all shore property owners.

Manuscript received May 27, 1971.

380 Transactions Illinois Academy of Science

Figure 6b. Corrugated area-way projections provide strength to erosion barrier

Roberts— Lake Shore Erosion

381

Figure 7

Gabion fabricated from heavy steel wire mesh.

Transactions Illinois Academy of Science

Figure 8. This gabion wall protects lake-front property

LOCAL EFFECTS OF THE NEW ACS CURRICULUM

SHELBA JEAN CHOATE MUSULIN AND BORIS MUSULIN

Department of Chemistry, Southern Illinois University, Carbondale, Illinois 62901

Abstract. — The questionnaire method was used to study high school background of and the effects of course prerequisites and content upon students in two suc¬ cessive fall terms of the freshman chem¬ istry class. The data were analysed with means and correlation coefficients, and statistical interpretations thereof. Well- known effects upon student performance were more relevant than the criticisms of a vocal minority.

In Fall, 1965, the Department of Chemistry of Southern Illinois Uni¬ versity, Carbondale, changed their undergraduate curriculum (South¬ ern Illinois University (1965)), in accordance with changes suggested in standards for accreditation of bachelor degree programs by the Committee on Professional Train¬ ing of the ACS (American Chemi¬ cal Society (1965)). The changes were the culmination of efforts by members of the ACS, along with highly qualified secondary school chemistry teachers, to develop two modern secondary school curricular plans (CHEMS, 1960; CBA, 1964). The ACS (1960) strongly stressed that a perspective science major should be prepared in one of these new curricular studies; thus, by 1965 the supposition that entering college science majors were better prepared seemed valid.

The new SIU freshman curricu¬ lum assumed that only a student without secondary school prepara¬ tion was deficient and could enter for credit the first course of the major sequence. Student com¬ plaints, the majority of which cen¬ tered upon this selection process, were of continuing concern. There¬ fore, the purpose of this research was to ascertain the suitability of the selection method and, in addi¬

tion, to ascertain certain student attitudes which were developed by the inauguration of the new pro¬ gram.

The Questionnaire

A questionnaire was sent to every student whose name appeared on the final class listing of the Fall, 1965 and Fall, 1966 second course classes. No mailing was made to Winter classes whose student popu¬ lation differed markedly, viz, no high school background, general science converts, and Fall failures. The first year mailing was made in the subsequent Winter quarter to a campus address. The returns which came over a prolonged time period included many rejections for insuffi¬ cient address therefore necessitat¬ ing many repeat mailings. Returns from the second year mailing, made to a home address during the holi¬ day period, came quickly with only two mail rejections, but with a smaller percentage return (70% vs. 63%). Total mailings were 402 and 398, respectively, with no follow-up mailing because of the high initial responses.

The questionnaire was divided into three parts: factual informa¬ tion concerning the student and his background; subjective opinions re¬ lated to background, instructors, and instructional materials; and subjective opinions related to per¬ formance level. The final part was to be answered only by those stu¬ dents who received a grade below C. The second year mailing differed in two respects: an added question categorizing, if possible, the stu¬ dent’s high school course as CBA, CHEMS, or other, and the addi-

383

384

Transactions Illinois Academy of Science

tion of Biology as a choice of col¬ legiate major. Other factors such as secondary school physics and mathematics which have been shown to influence collegiate chemistry grade performance (Hadley, et al. (1953)) were not investigated.

In the treatment of data, each response in each item was given a numerical value and all responses for one student placed on a single punched card. A missing data cor¬ relation program (Wright (1962)) was run on an IBM 1620 digital computer with 40K storage and a PR 025 monitor. This program yields the mean of each variable, the standard deviation of each vari¬ able, and the bivariate (Pearson) correlation coefficient (Baten (1938)) for each variable pair.

Sample Profile

The profile of the average stu¬ dent, constructed from an analysis of variable means, for each class is presented in Table 1. Response spread may be found from one stan¬ dard deviation of each item. The difference between the means in Table 1 are all significant at more

than the 1% level (Baten (1938)). A few items are compared to re¬ plies obtained in a one-year survey taken by the Department of Chem¬ istry (1965). Briefly the means in¬ dicate that the two student sam¬ ples are very similar but with the more recent class containing a some¬ what higher quality (academically) student from a more populated county (geographically, more cen¬ tral) of the state. This improvement in academic quality is to be ex¬ pected from the national trend of rising admission standards.

Comparison to a sample profile of the entire freshman class (Cham- berlein (1965, 1966)) indicates that the average chemistry student is more likely to be a male, to major in a science-related field, to be above average in academic quality, and to come from a small secondary school in the immediate geographic region of the university. The mean high school chemistry background reflects the time-lag in curriculum change in these smaller, regional secondary schools and would indi¬ cate a high probability that the

Table 1. A Profile of the Average Introductory Chemistry Student

Item

1965-6

1966-7

Sex (% Males)

89

85

61*

58*

Age (Years)

19.3

19.3

College Credit Earned (Quarter Hours)

34.2

41.0

Percent with Academic Scholarship

28

32

Percent Working

40

44

Ill. High School Metropolitan

40.5

54.2

Area (In Kilopeople)

72.1*

72.9*

Size of High School

196

212

High School Rank

68.2

73.5

(Percentile)

60.0*

65.2*

Years of High School Chemistry

1.15

1.18**

1.02

High School Chemistry Average (A = 5.00)

3.80

3.86

Attendance in other non-High Chemistry (Percent)

30

30

Grade Average in Introductory College

Chemistry (A = 5.00)

2.57

2.87

*Entire Freshman Class

* independent Survey of Introductory Chemistry Class

Musulin and Musulin — Effects of the New ACS Curriculum

385

average freshman chemistry stu¬ dent might be deficient in modern high school chemistry. The slightly higher 1965 mean is explained by an abnormal influx of Oriental stu¬ dents as a result of a revised immi¬ gration statute. The mean credit hours earned indicate a sizeable number of non-first term freshman and those with previous collegiate chemistry creating a heterogeneity contrary to a goal sought by the curriculum changes.

Correlation Ranking

Table 2 presents the correlation coefficients between the student's grade performance and each factor affecting grade performance. The table is divided into objective and subjective sections and, within each section, the factors are ordered ac¬ cording to decreasing importance. Individual items are grouped ac¬ cording to approximate level of sig¬ nificance, e.g. 0.16 is significant at the 1% level and 0.12 at the 5% level (Baten (1938)) for these sam¬ ple sizes.

The results show that the most important factor affecting the stu¬ dent’s collegiate grade was his grade

performance in high school chem¬ istry. Less than 10% of the students responding claimed to have had either CBA or CHEMS which may be the reason for the relatively minor role of high school size and location. Overall the type of back¬ ground is not as important as the amount of background, and the background factors are not as im¬ portant as the academic quality of the student. Table 2 also indicates that the better student felt better prepared but neither hours of study nor text opinion had any signifi¬ cant effect on the student’s grade performance. The students tended to report a low number of hours of study (3 - 7) relative to the rule of thumb (two hours study per credit hour, i.e., 10) quoted to the stu¬ dents. Uniformly, all students re¬ ported an unfavorable impression of the textbook.

Table 3 (in the same format as Table 2) which correlates the stu¬ dent’s opinion of his background to various factors again illustrates the most important factor is final grade performance. Maturity factors are significant at the 1% level indicat¬ ing an increasing ability to judge

Table 2. Correlation Coefficient Ranking of Factors Affecting the

Student’s Grade Performance

Item

1965-6

1966-7

Objective Items

High School Chemistry Average

.402

.410

High School Class Rank

.333

.384

Collegiate Major

.235

.166

Years of High School Chemistry

.234

.222

Illinois High School Location

.133

.251

Age

.158

.090

Size of High School

.149

.082

Year in College

.113

.099

Prior Years of College Chemistry

.095

.026

Sex

.044

.065

Type of Instructor

.089

.007

Type of High School Course

• • • •

.005

Subjective Items

.379

Background

.354

Opinion of Instructor

.262

.223

Opinion of Graduate Assistant

.228

.101

Hours of Study Per Week

.126

.091

Opinion of Text

.065

.110

386

Transactions Illinois Academy of Science

background. The better prepared student studies a lesser number of hours which creates an inherent bias in that coefficient.

The Below Average Group

In both years, the poorly per¬ forming student felt slightly supe¬ rior, in intellectual ability, to his peers. This tendency manifested it¬ self to a greater degree in the more recent class whose overall academic quality was superior. Contrary to the directions, many selected mul¬

tiple reasons in appraising their poor performance with not enough study as the principal cause. Lack of interest and/or poor background were choices within one standard deviation of the mean. Consequent¬ ly, in a random sample (110 and 80, respectively) of poor students, poor background does not emerge as their primary reason for poor per¬ formance. Poor collegiate instruc¬ tion was not significant.

The 1965 class best correlated poor background to the nature of

Table 3. Correlation Coefficient Ranking of Factors Affecting the

Student’s Opinion of Background

Item

1965-6

1966-7

Objective Items

Grade in College Chemistry

.354

.379

Age

.212

.157

Year in College

.174

.176

Prior Years of College Chemistry

.171

.137

Illinois High School Location

.193

.068

Years of High School Chemistry

.108

.116

High School Chemistry Average

.162

.063

Size of High School

.086

.066

Sex

.073

.059

Collegiate Major

.047

.053

High School Class Rank

.027

.053

Type of High School Course

.005

Subjective Items

Hours of Study Per Week

.189

.262

Opinion of Graduate Assistant

.143

.006

Opinion of Instructor

.009

.152

Opinion of Text

.049

.060

the material given by a particular lecture instructor (for the purpose of judging correlation coefficients, a subjective judgment was made on the amount of theoretical material used by each lecturer in the fresh¬ man course). That is, the poorer students in the sections which em¬ phasized theoretical material felt their lack of background to a great¬ er degree than the poorer students in other sections. The significance was approximately at the 1% level. For the 1966 class, the correlation coefficient (.116) was nonsignificant for the sample size thus precluding the same conclusion.

Summary

This study shows that the stu¬ dent’s academic quality is far more important than any other single factor. The degree of importance is great enough to override limitations imposed by a possible non-normal distribution in the student sample with respect to secondary school background. Collegiate chemistry departments need not be greatly concerned over the type of back¬ ground because the amount of sec¬ ondary school chemistry is more important than the type of such chemistry.

Musulin and Musulin — Effects of the New ACS Curriculum

387

Student sampling indicated that the most vociferous comments orig¬ inated from a minority grouping.

Acknowledgement

Partial financial support was given by the Department of Chemistry of South¬ ern Illinois University, Carbondale. Gra¬ tis use of the computing facilities was given by the Data Processing and Com¬ puting Center of Southern Illinois Uni¬ versity, Carbondale.

Literature Cited

American Chemical Society. 1960. Chemistry and Your Career. A Voca¬ tional Guidance Pamphlet. Washing¬ ton, D. C.

_ , 1965. Minimum Stan¬ dards Used as Criteria in Evaluating Undergraduate Professional Education in Chemistry. Pamphlet. Committee on Professional Training. Washington, D. C.

Baten, W. D., 1938. Elementary Math¬ ematical Statistics. John Wiley & Sons, Inc. New York, N. Y.

Bailer, Jr. J. C., Moeller, T., and Kleinberg, J., 1965. University Chemistry. D. C. Heath and Company, Boston.

CBA. 1964. Chemical Systems. Web¬ ster Division, McGraw-Hill Book Com¬ pany, New York, N. Y.

Chamberlin, L. J., 1965. Freshman Profile. Southern Illinois University, Carbondale, Illinois. Fall. (Mimeo¬ graphed.)

_ _ _ , 1966. Freshman Profile.

Southern Illinois University, Carbon¬ dale, Illinois. Fall. (Mimeographed.)

Chems. 1960. Chemistry. W. H. Free¬ man. San Francisco, California.

Department of Chemistry. 1965. Summary of Students in 111b, Fall 1965: High School Background and Majors. Carbondale, Illinois. (Type¬ script.)

Hadley, E. H., Scott, R. A., and Van

Lente, K. A., 1953. The Relation of High-School Preparation to College Chemistry Grades. J. Chem. Educ., 30:311-313.

Southern Illinois University. 1965. Bulletin. Vol. 7, no. 10, Carbondale, Illinois. October, p. 66.

Wright, W. E., 1962. Southern Illinois University Data Processing and Com¬ puting Center Missing Data Correla¬ tion Program for Symmetric and Asym¬ metric Matrices for the IBM 1620. Carbondale, Illinois. (Mimeographed.).

Manuscript received April 26, 1971.

THE STABILIZATION OF A GULLY BY NATURAL

FOREST SUCCESSION

ROBERT A. BULLINGTON

Department of Biological Sciences , Northern Illinois University

De Kalb, Illinois, 60115

Abstract. — In an abandoned pasture in Rockvale Township of Ogle County, Illinois, there is an old gully formed dur¬ ing a period of cultivation between 1835 and 1900. It is about 1300 feet long, varies in width from five to 100 feet and ranges to a depth of 15 feet. A photo¬ graph of 1904 shows a few scattered clumps of small trees in the gully. Today it is obscured by many mature trees and abundant undergrowth. A survey of gully trees in 1958 revealed box elder to be the most abundant, with black walnut, red elm, and American elm following. In 1969, red elm and black walnut shared dominance, with hackberry, black oak, box elder, black cherry and American elm next. Most of the older box elder were dead. The American elm had increased substantially in total number, but one- half were dead of Dutch elm disease. Most red elm were unaffected by the dis¬ ease. The number of tree species in¬ creased from 13 to 21 during the years 1958 to 1969. Many new species of forest- related herbaceous plants had appeared. It is evident that in a period of about 70 years the gully has become stabilized and revegetated with numerous forest species and is developing rapidly into a mature deciduous forest condition.

Few ecological studies of secon¬ dary succession give any attention to the development of natural vege¬ tation in gullies. Most are con¬ cerned with abandoned agricultural land. In some studies the recovery of a gully may be mentioned inci¬ dentally.

Oosting (1942) reported that, in the Piedmont, natural vegetation of gullies is very slow if head ero¬ sion continues. Eventually, the typ¬ ical herbs and legumes of the old field appear. Marks (1942) also re¬ ported slow recovery of gullies in Wisconsin, with local weeds fol¬ lowed by perennial grasses and herbs. Early woody vegetation con¬ sisted of Rubus, Ribes, and Prunus

virginiana with elder ( Sambucus ) and black locust ( Robinia ) develop¬ ing on the upper gully slopes. In a gully thicket in Michigan, Brewer et al. (1969) found black locust a dominant tree associated with var¬ ious shrubs, vines, and herbs.

Since no detailed analysis of the vegetation of a stabilized gully has been found in the literature, the author will attempt to show what has happened in recent decades in a northern Illinois setting.

Description of Study Area

A detailed account of the study site has previously been reported (Bullington, 1970). Only a brief de¬ scription will be included here. The main features are shown in Figure 1.

An old pasture of approximately 30 acres is located on property known as The Stronghold, owned by the Presbyterian Camping As¬ sociation of Northern Illinois. The field is in the center of section 33 of Rockvale Township (T 24 N., R 10 E.), Ogle County, Illinois. The site is on a hillside to the west of the Rock River Valley extending westward from Illinois Highway 2, north of the town of Oregon.

The gully is the dominant feature of the old pasture, extending from the center to the eastern edge, a distance of about 1300 feet, and dropping from an elevation of about 780 feet at the west end to 710 at the east. At the gully head it rapidly deepens to about 15 feet below the adjacent ground level and varies in width from a few to about 30 feet. As it progresses to less steep terrain, it becomes quite shallow and broadens to about 100 feet.

388

Bullington — Gully Succession

389

Figure 1. The study area at The Stronghold, Oregon, Illinois.

Portions of the upper part are cut down to bedrock of Platteville limestone, while the lower part has considerable areas of deposited silt and debris. At various places, the gully has been partially filled with old fence wire, farm machinery, and other debris. Little visible erosion has occurred in the past 20 years except for some shallow channels cut in the sediments of the lower part of the gully.

The hillside field was cleared of forest and cultivated by pioneers starting soon after 1835. Decades of cultivation resulted in extensive erosion of the field, with removal of all topsoil and the formation of the large central gully with a few small branches. Sometime before 1900, the field was seeded to bluegrass (and perhaps other grasses and le¬ gumes) and used for pasture until 1948. No domestic livestock has dis¬ turbed the field since. The entire field has undergone rapid secon¬ dary succession since 1948, with distinct competition between forest and prairie vegetation (Bullington, 1970).

A photo published in 1904 shows the gully quite clearly with a few clumps of small trees in limited areas. By the time of an aerial photo of 1939, there was a distinct cover of trees present over most of the gully.

Procedure and Observations

The author’s first observation of the gully vegetation was in 1958 (Table 1). In all there were 551 trees counted, with box elder ( Acer negundo) accounting for 295, or 53.5%. The likely seed source was box elders along the old roadway at the foot of the gully, a few of which were present in the 1904 photo. They are uncommon in the forest both to the north and the south of the field. Everywhere in the field, however, box elder is prom¬ inent in the new growth. Evidently the seeds can be dispersed a con¬ siderable distance, and the seed¬ lings develop readily in an open area. The box elder is a common pioneer tree.

The 1958 survey showed that wal¬ nut ( Juglans nigra) was next in

390

Transactions Illinois Academy of Science

Table 1. Trees found in the gully of Strong field by reconnaissance survey in summer of 1958.

Species

Number

Percent of Total

Acer negundo

295

53.5

Juglans nigra

95

17.2

Ulmus rubra

35

6.4

Ptelea trifoliata

23

4.2

Ulmus americana

18

3.3

Populus grandidentata

17

3.1

Malus ioensis

16

2.9

Carya ovata

15

2.7

Crataegus mollis

11

2.0

Prunus serotina

10

1.8

Quercus velutina

9

1.6

Celtis occidentalis

6

1.1

Prunus americana

1

.2

Total

551

100.0

abundance with 95 trees, or 17.2%. Shagbark hickory ( Carya ovata) and black oak ( Quercus velutina) were represented by a small number of trees. Both could have been intro¬ duced from the mature forest about 200 feet to the north by the activity of squirrels.

Increment borer cores were taken of the larger trees in the gully in 1959 and 1963 (Table 2). Cores were collected and studied from walnut, box elder, American and red elms ( Ulmus americana and U . rubra), black oak, large-toothed aspen ( Populus grandidentata) , and black cherry ( Prunus serotina). The trees ranged in estimated age from 25 to 40 years. It was determined that they became established in the years between 1914 and 1933, a period when the field was used for pastur¬ ing and hay meadow. It is not known whether the trees present in the 1904 photo were still alive when the check on age was made.

Between 1958 and 1969, the gully was visited frequently at various seasons. The presence of new spe¬ cies of plants was often noted but no statistical information was gath¬ ered. During the eleven-year period the gully vegetation matured and took on a more forest-like aspect.

It was obvious that new forest spe¬ cies of herbs and woody plants were becoming established.

In October of 1969 a detailed survey of the trees of the gully was undertaken. Because there was no uniformity in distribution of spe¬ cies, a complete count of all trees was made. Starting at the upper west end, ten-feet wide strips were marked off with string. Not only were the trees of the gully included, but also those to a distance of ten feet on each side of the banks. This was roughly the point of division between the adjacent old-field vege¬ tation and that of the eroded gully.

Trees were tallied by species and by size classes. Size was measured in inches of diameter at four and a half feet above the ground (dbh). Five size classes were arbitrarily selected (under 1 inch, 1-6 inches, 6-9 inches, 9-12 inches, and over 12 inches). Separate records were kept of the west one-half and the east one-half because of differences in both terrain and species.

Because of considerable mortality among a few species, dead trees were recorded. Numerous Ameri¬ can elms were dead from the Dutch elm disease. This was not present in the earlier survey of 1958. Many

Bullington— Gully Succession

391

Table 2. Age estimates of gully trees by increment borer samples at 4-1/2 feet height.

Year Taken in October

Species

Estimated Age at 4-1/2 Feet

Estimated Date of Establishment

1959

Juglans nigra

40

1914

1959

Ulmus americana

30

1924

1959

Acer negundo

30

1924

1963

Quercus velutina

40

1918

1963

Ulmus americana

35

1923

1963

Juglans nigra

30-35

1923-28

1963

Ulmus americana

30

1928

1963

Populus grandidentata

30

1928

1963

Quercus velutina

30

1928

1963

Acer negundo

25-30

1928-33

1963

Ulmus americana

25-30

1928-33

1963

Prunus serotina

25

1933

1963

Ulmus rubra

25

1933

box elders were also dead, presum¬ ably from shading and age.

The results of the 1969 survey are given in Table 3. Thirty percent of the total were under one inch dbh. Over 53% were between one and six inches. New trees were much more abundant in the west half of the gully, where 17 species were in the smallest size category and 15 in the second group. In the east half, there were 11 and 10 spe¬ cies in the two smallest size groups. The west half of the gully has more of the appearance of a forest, with a well-developed crown and open understory. The east half, much broader and with more silt deposits, is quite open, with more scattered trees and a heavy ground cover of brambles and vines which are al¬ most impenetrable in places. Seed¬ ling trees have a better chance of surviving in the west half.

The total number of species in 1969 was 21. Only 13 were recorded in 1958. All but one of the new spe¬ cies of 1969 were represented by young trees under 6 inches dbh. The exception was white oak ( Quercus alba) with two trees between 6 and 12 inches. They were located along the margin where they were not counted in 1958.

Abundant new reproduction has occurred with red elm, black oak, hackberry (< Celtis occidentalis) , and the black cherry in the west end only. There are mature trees of these species for seed sources in the west end but not in the east end, except for one red elm. On the other hand, the black walnut is reproduc¬ ing heavily in both areas. Mature trees are abundant throughout the gully, but there are more in the east end where walnut reproduction is the heaviest. It is likely that a por¬ tion of the lower east end of the gully will eventually develop into a dominant black walnut forest.

In order to compare reproduction in the gully with that in a mature forest on the north side of the old field, only a distance of 300 feet from the gully at the east end, and 100 feet at the west end, a count was made of the trees in a marginal 50-feet strip running parallel with the gully. The north forest strip is 1000 feet long and is the fence-row margin of an extensive mature up¬ land oak forest, probably of the type that was cleared from the old- field area in the 1830’s. The north strip was assumed to be close enough to have had significance as an early seed source for the gully.

Table 3. Number of trees in gully by size classes (Oct. 1969).

392

Transactions Illinois Academy of Science

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Note: Dead trees of three species are not included in table totals. Numbers not totalled are italicized.

Bullington— Gully Succession

393

Table 4. Trees of gully in order of abundance in 1969 compared with 1958

and with rank in adjacent areas.

No. in Gully 1969

Species

Rank

Gully

1969

Rank

Gully

1958

Ran Old 1

k in Field

Rank in Forest 1969

Rank in Old Road 1970

1959

1970

257

Ulmus rubra

1

3

3

2

3

4

244

Juglans nigra

2

2

2

1

8

2

113

Celtis occidentalis

3

12

5

3

83

Quercus velutina

4

11

5

4

2

79

Acer negundo

5

1

1

4

9

1

77

Prunus serotina

6

10

10

11

7

4

66

Carya ovata

7

8

9

10

1

8

64

Ulmus americana

8

5

7

9

4

30

Populus grandidentata

9

6

11

7

6

23

Malus ioensis

10

7

6

3

9

19

Prunus virginiana

11

7

10

Juniperus virginiana

12

6

Crataegus mollis

13

9

4

8

5

Fraxinus americana

14

2

Acer saccharum

15

2

Quercus macrocarpa

2

Quercus alba

12

4

2

Prunus americana

13

7

6

2

Ptelea trifoliata

4

1

Viburnum lentago

1

Catalpa speciosa

21

1088

Total Species Present

21

13

11

12

9

9

The same size classes were used in the forest count as in the gully. The survey was also made in Oc¬ tober of 1969. The details of this survey will be reported in another paper. Brief mention will be made here of pertinent information. Black oaks and the shagbark hickory were discovered to be the dominant trees of this forest. Most of the recent reproduction was hickory. There were few walnuts.

Table 4 compares the various areas studied in terms of rank order based on total numbers of each spe¬ cies. There were only nine species in the 50-feet strip at the edge of this forest. This pattern continues through the forest. Table 4 also compares the gully trees with those of the adjacent old field to the north recorded in two quadrat surveys in 1959 and 1970. This narrow strip of former pasture separates the gully from the forest. It varies from 100 to 300 feet in width, with the nar¬

row end to the west. Its principal species were the black walnut and red elm. The strip had 11 species in 1959 and 12 in 1970.

Also compared with the gully is a 420-feet strip of abandoned road running at right angles to the east end of the gully. This old gravel road has been undisturbed since 1922 except for a mowed path down the center. Large trees now arch overhead. The old road right-of- way is approximately 60 feet wide. A count of the road trees in 1970 showed that box elder was the most abundant. Except for the absence of the black oak, the top five spe¬ cies of the roadway were the same group as the top six of the gully, although the rank order was differ¬ ent (see Table 4).

No detailed analysis of shrubs and vines has been made, although they are now a prominent part of the ground cover in portions of the

394

Transactions Illinois Academy of Science

gully. The 14 species identified are listed in Appendix I.

As the forest of the gully ma¬ tures, there is a steady invasion of herbaceous plants, particularly the spring ephemerals. The 50 species identified to date are listed in Ap¬ pendix II.

Species common to disturbed areas, such as stinging nettle ( Ur- tica dioica ) and black snakeroot (, Sanicula gregaria) have been com¬ mon since the first observations were made. Species of rich woods, such as rattlesnake fern ( Botrychium virginianum) , purple trillium ( Tril¬ lium recurvatum), may apple ( Po¬ dophyllum peltatum), and Solomon’s Seal ( Polygonatum biflorum) have entered recently.

Of special interest are the two orchids found in the gully, each in the type of habitat described for it in Gray’s Manual of Botany (Fer- nald, 1950). The broad-leaved tway- blade (. Liparis lilifolia) was found in 1970 in the more open, drier, up¬ per part of the gully. It had been established nearby for several years in a young thicket of trees in the old field.

The showy orchis (< Orchis specta- bilis) was first reported about 1958 in a moist wooded area across the old roadway from the east end of the gully. By 1970 it was abundant in the more mature central portions of the gully. Fell (1955) reported the two orchids growing together in a woods in adjacent Winnebago County.

Most herbaceous species listed in Appendix II are in the type of hab¬ itat and associated with the typi¬ cal tree species reported by Swink (1969) for northern Illinois counties.

Discussion

The eleven-year period from 1958 to 1969 was a time of rapid change and development of the gully vege¬ tation. The area had progressed

from an open gully in 1900, still subject to erosion, to a fairly stable deciduous forest situation in about seven decades.

Some distinct changes have oc¬ curred in the species of trees. A few were disappearing. The wafer ash (. Ptelea trifoliata), number four in abundance in 1958, dwindled to two specimens in 1969. Heavy shad¬ ing no doubt accounted for its de¬ mise. It is still common in the open parts of the old field.

Box elder, a common pioneer tree dominant in the old field and also in the gully in 1958, was rapidly dying. Most of the box elders over six inches dbh were dead. The smaller specimens recorded were mostly sprouts from the base of dead or dying trees.

The American elm is dying from the Dutch elm disease, with about one-half of the trees dead. It is de¬ creasing in rank, going from 5th to 8th, but the actual percentage of American elm trees is increasing (3.3% to 5.9%). There is still active reproduction of this species.

Red elm is reproducing vigor¬ ously, especially in the west end of the gully. It has moved from third rank with 6.4% to first place in 1969 with 23.6% of the total of 1088 live trees in the gully. Only two large trees were found dead. Apparently at this midpoint in suc¬ cession, red elm is an important species. So far, it does not seem very susceptible to the disease that is decimating the American elm.

Downy haw ( Crataegus mollis) is not surviving well in the shade of the gully. The few remaining speci¬ mens are quite small. Wild crab apple ( Malus ioensis), a common pioneer associated with the haw, is still a common understory tree in the gully. However, it ranked in tenth place there in 1969 compared to a third place rank in the adjacent field. It was absent from the adja-

Bullington — Gully Succession

395

cent mature forest sample. As shad¬ ing increases, the haw and crab apple will probably disappear from the gully.

The increase in some tree species is quite marked. Hackberry has moved from near the bottom of the 1958 list to third place in 1969, with 10.4% of the total trees. There is very heavy reproduction of the hackberry. However, the saplings are being damaged or killed at a rapid rate by winter girdling by rabbits.

Black oak has assumed a promi¬ nent place in recent years. It is co¬ dominant with hickory in the near¬ by forest and shows signs of assum¬ ing this same place in the gully, at least in the west half. Hickory is also increasing in prominence. There is active reproduction of both spe¬ cies.

Black walnut, actively reproduc¬ ing and accounting for 22.4% of the total trees in 1969, is a rather sur¬ prising contender for future domi¬ nance of the gully forest. It is rather infrequent in the mature forest. However, it is known that mature trees have been selectively logged from the forest in the past. Black walnut is known to have an inhibit¬ ing effect upon other species. Per¬ haps it is the presence of walnut that is hastening the death of box elder. The two grow in the same habitat. Another species that is in¬ creasing in abundance is the black cherry. A few mature trees provide the abundant seeds for prolifera¬ tion of this tree.

The improvement of the gully environment as a habitat for forest trees is indicated by the rapid in¬ crease in total species to 21. No other area sampled has more than a dozen species. Some of the re¬ cently invading species are prob¬ ably random or accidental seedings of no significance. Catalpa {Catalpa speciosa) is not likely to multiply,

and the two specimens of burr oak ( Quercus macrocarpa) may be the only ones that enter. Red cedar (. Juniperus virginiana) has been spread from open-grown trees in the old field. It will not do well in the shade. The newly reported choke cherry ( Prunus virginiana) may have been overlooked in 1958. It will not be more than an understory tree.

Two new species are of special interest, even though their numbers are small. Two sugar maple ( Acer saccharum) saplings have appeared. This tree of the climax deciduous forest is abundant in a forest valley only a few hundred yards south of the gully. Also from the forest val¬ ley have come at least five young white ash ( Fraxinus americana). Four specimens have passed the 1 inch dbh mark and show promise of future development.

It should be noted that nine of the ten most abundant trees in the gully are also among the most com¬ mon invaders of the adjacent blue- grass old field. The exception is hackberry, which is absent from the field. Two species common as pioneers in the old field, downy haw and wild plum ( Prunus americana) are rare in the gully. As noted be¬ fore, shading is probably the limit¬ ing factor.

Conclusion

In six or seven decades, an open gully has become stabilized by nat¬ ural vegetation and has progressed to a mid-successional stage of de¬ ciduous forest development. There is active reproduction of numerous tree species. Many herbaceous plants, typical of rich deciduous woods, have become established under the mature trees. There is evidence that the rate of develop¬ ment is increasing. A few more de¬ cades may result in a near-climax

396

Transactions Illinois Academy of Science

deciduous forest in a stabilized con¬ dition.

The observations of this study have implications for gully manage¬ ment in deciduous forest areas. Planting an exotic species is a com¬ mon practice in the management of waste areas such as gullies. It is suggested that such places be per¬ mitted to develop naturally through the processes of secondary succes¬ sion, especially if good seed sources are in the vicinity.

Two circumstances that have ex¬ isted in the Strong old-field situa¬ tion are no doubt of considerable importance. The eroded field sur¬ rounding the gully was planted to grass, thus stabilizing the soil of the area and reducing runoff of wa¬ ter. At a later time, livestock were excluded from the area. With these aids in management, natural pro¬ cesses have succeeded.

Literature Cited

Brewer, R., A. Raim, and J. D. Robins.

1969. Vegetation of a Michigan grass¬

land and thicket. Occas. Papers of the C. C. Adams Center for Ecolog. Stud¬ ies. W. Mich. Univ., Kalamazoo. 18: 1-29.

Bullington, R. A. 1970. Competition between forest and prairie vegetation in twenty years of secondary succession on abandoned land in Ogle County, Illinois. Proc. Symp. on Prairie and Prairie Restoration, Knox Coll., Gales¬ burg, Ill. 20-23.

Fell, E. W. 1955. Flora of Winnebago County, Illinois. Nat. Cons., Washing¬ ton, D. C. 1-207.

Fernald, M. L. 1950. Gray’s Manual of Botany, 8th Ed. American Book Co., New York. 1-1632.

Marks, J. B. 1942. Land use and plant succession in Coon Valley, Wis. Ecol. Mono. 12 (2) :1 13-133.

Oosting, H. J. 1942. An ecological anal¬ ysis of the plant communities of the Piedmont, N. Carol. Am. Midi. Natur. 28:1-126.

Swink, F. 1969. Plants of the Chicago region. The Morton Arboretum, Lisle, Ill. 1-445.

Manuscript received June 25, 1971.

Contribution No. 508 from the Depart¬ ment of Biological Sciences, Northern Il¬ linois University.

Appendix I. Shrubs

Berberis Thunbergii DC Lonicera prolifera (Kirchn.) Rehd.

Lonicera sp.

Parthenocissus quinquefolia Planch. Rhamnus cathartica L.

Rhus glabra L.

Rhus radicans L.

Ribes missouriense Nutt.

Rubus allegheniensis Porter Rubus occidentalis L.

Sambucus canadensis L.

Smilax tamnoides L. hispida (Muhl.) Fern. Viburnum trilobum Marsh.

Vitis riparia Michx.

and Vines of Gully

Japanese barberry grape honeysuckle honeysuckle (ornamental) Virginia creeper common buckthorn smooth sumac poison ivy wild gooseberry blackberry black raspberry common elderberry bristly greenbrier highbush cranberry riverbank grape

Bullington — Gully Succession

397

Appendix II. Herbaceous Plants of Gully

Note: The identification of the herbaceous plants of the gully is an on-going project. Some of the plants listed are tentatively identified and require examination at other times of the year than when seen, especially at flowering time. No attempt has been made to sort out the species of certain genera, as Aster and Solidago. On each visit to the gully, new species are discovered and no doubt there will be new ones found in the future. Some plants have undoubtedly been missed. As the environment changes, there will be new invaders for future discovery.

tall agrimony common ragweed great ragweed wild columbine common burdock blunt-leaved sandwort Jack-in-the-pulpit aster lady fern yellow rocket rattlesnake fern black mustard pale Indian-plantain sedge tick-trefoil shooting-star common boneset Joe-pye-weed wild strawberry bedstraw wild licorice small bedstraw white avens wild cranesbill sunflower Virginia waterleaf Jewelweed

broad-leaved twayblade Virginia bluebells wild bergamot showy orchis hairy sweet cicely parsnip

common plantain mayapple Solomon’s seal kidneyleaf buttercup bristly buttercup golden-glow black snakeroot late figwort golden ragwort false spikenard starry false Solomon’s seal carrion flower goldenrod common dandelion purple meadow-rue purple trillium wild coffee stinging nettle smooth yellow violet common blue violet

Agrimonici gryposepala Wallr.

Ambrosia artemisiifolia L.

Ambrosia trifida L.

Aquilegia canadensis L.

Arctium minus (Hill) Bernh.

Arenaria laterifolia L.

Arisaema triphyllum (L.) Schott Aster sp.

Athyrium sp.

Barbarea vulgaris R. Br.

Botrychium virginianum (L.) Sw.

Brassica nigra (L.) Koch Cacalia atriplicif olia L.

Carex pennsylvanica Lam.

Desmodium glutinosum (Muhl.) Wood (?) Dodecatheon meadia L.

Eupatorium perfoliatum L. (?)

Eupatorium purpureum L.

Fragaria virginiana Duchesne Galium aparine L.

Galium circaezans Michx.

Galium trifidum L.

Geum canadense Jacq.

Geranium maculatum L.

Helianthus sp.

Hydrophyllum virginianum L.

Impatiens sp.

Liparis lilifolia Richard Mertensia virginica (L.) Pers.

Monarda fistulosa L.

Orchis spectabilis L.

Osmorhiza Claytoni (Michx.) C. B. Clarke Pastinaca sativa L.

Plantago major L.

Podophyllum peltatum L.

Polygonatum biflorum (Walt) Ell. Ranunculus abortivus L.

Ranunculus hispidus Michx. (?)

Rudbeckia laciniata L.

Sanicula gregaria Bickn.

Scrophularia marilandica L.

Senecio aureus L. (?)

Smilacina racemosa (L.) Desf.

Smilacina stellata (L.) Desf.

Smilax lasioneura Hook.

Solidago sp.

Taraxacum officinale Weber Thalictrum dasycarpum Fisch. & Lall. Trillium recurvatum Beck Triosteum perfoliatum L.

Urtica dioica L.

Viola pennsylvanica Michx.

Viola papilionacea Pursh

A NEW LOCALITY AND A NEW HOST RECORD FOR TRICHODINA DISCOIDEA DAVIS, 1947 IN SOUTHERN ILLINOIS

LAWRENCE J. BLECKA

Department of Zoology , Southern Illinois University, Carbondale 62901

Abstract. — Trichodina discoidea is reported from Lepomis cyanellus in Jack- son County Illinois. This report marks the occurrence of this peritrich on a new host and in a new geographic area. T. discoidea was also found onL. macrochirus and Ictalurus punctatus in Jackson Coun¬ ty.

Davis (1947) named T. discoidea from material collected at Kearneysville, West Virginia. At that location T. discoidea was found on the gills of the bluegill, Lepomis macrochirus, the black crappie, Pomoxis sparoides, and the rock bass, Amhloplites rupestris. Davis also noted the occurrence of T. discoidea on the gills of the channel catfish, Ictalurus punctatus taken from the Mississippi River near Fairport, Iowa.

From July-December, 1970, fish from a commercial hatchery near Gorham, Jackson County, Illinois were collected by seining and transported to the labora¬ tory alive for examination. Captive fish were segregated into species and carried in 19 1. plastic bags containing 3-4 1. of oxygenated well water. All specimens were examined for ectoparasites within 36 hr. Before examination the fish were double pithed, the opercula, fins, tail, and body were placed in separate dishes of distilled water, and were observed un¬ der stereomicroscope at 30-60X. Tricho- dinids were prepared for silver impregna¬ tion by smearing the host material on a clean glass slide. The slides were air dried and stained after the method of Lorn (1970). Infected gills were also sectioned and stained with hematoxylin and eosin

after Humason (1967). Identification and biometric characteristics followed the cri¬ teria of Lorn (1958). Microscopy was with a Ziess photomicroscope.

During the study the occurrence and incidence of T. discoidea was noted on 17.6% of 17 L. macrochirus, 18.1% of 22 L. cyanellus, and 21.0% of 93 I. punctatus. The parasite occurred only on the gills of these hosts. This is the first report of T. discoidea from southern Illinois, and the first time the parasite has been re¬ ported from the green sunfish,L. cyanellus.

T. discoidea has now been reported from four centrarchids as well as the channel catfish which would seem to in¬ dicate a rather broad host specificity. Presently additional studies into the par¬ asite’s host specificity, host induced vari¬ ation, and biology are underway.

Literature Cited

Davis, H. S. 1947. Studies of the pro¬ tozoan parasites of freshwater fishes.

U. S. Dept. Interior Fishery Bull. #41, 1-29.

Humason, G. L. 1967. Animal Tissue Techniques. 2nd ed. W. H. Freeman, San Francisco, 569 p.

Lom, J. 1958. A contribution to the sys- tamatics and morphology of endopara sitic trichodinids from amphibians, with a proposal of uniform specific charac¬ ters. J. Protozool. 5:251-263.

_ _ _ _ _ .. 1970. Observations on

trichodinids ciliates from freshwater fishes. Arch. Protist. 112:153-177.

Manuscript received April 22, 1971.

398

A NEW DISTRIBUTION RECORD FOR LYCOPODIUM FLABELLIFORME IN ILLINOIS

LOY R. PHILLIPPE

Eastern Illinois University, Charleston, Illinois

On September 20, 1970, a clubmoss, Lycopodium flabelliforme (Fern.) Blanch, was collected near Ducanville (Sect. 25, T6N, R12W) in Crawford County, Illi¬ nois. This appears to be the third county in the state in which a naturally occurring population of this species has been found. The first native station was found in Pope County near Lusk Creek (Jones & Fuller, 1955) and the second in Ogle County near Muir Creek (Mohlenbrock, 1967). The Crawford County specimen is about 100 miles north of the Lusk Creek station and over 200 miles south of the Ogle County station. In Indiana, Deam (1940) records this clubmoss for 7 counties. The nearest site is in Martin County, about 50 miles east of this recent Illinois find. Three adventive stations of this species have also been reported from northern Illinois by Mohlenbrock (1967).

The habitat in Crawford County for the Lycopodium flabelliforme population is a lowland terrace thicket of second growth woods near a branch of Brushy Creek. The small clump of clubmoss is growing at the base of a 15-foot tall red oak. The dominant trees of the area, which are all under 20 ft. tall, are river birch and sugar maple and to a lesser ex¬ tent black cherry, sassafras, sycamore,

shingle oak, and red oak. The understory shrubs consist mostly of Campsis radi- cans (L.) Seem, and Rubus spp. The her¬ baceous plants of the immediate area consist of Asplenium platyneuron (L.) Oakes, Onoclea sensibilis L., Cinna arun- dinacea L., Muhlenbergia sobolif era (Muhl.) Trim, Glyceria striata (Lam.) Hitchc., Panicum latifolium L., Agri- monia parvifiora Ait., Galium aparine L., Prunella vulgaris L., Aster pilosus Willd., Eupatorium perfoliatum L., Eupatorium rugosum Houtt., and Solidago caesia L.

A small portion of one plant (Phillippe #68) is preserved in the herbarium of Eastern Illinois University.

Literature Cited

Deam, C. C. 1940. Flora of Indiana. Wm. B. Burford Printing Co., Indian¬ apolis. 1236 pp.

Jones, G. N., and G. D. Fuller. 1955. Vascular Plants of Illinois. University of Illinois Press, Urbana. xii+593 pp.

Mohlenbrock, R. H. 1967. The Illus¬ trated Flora of Illinois. Ferns. South¬ ern Illinois University Press, Carbon- dale. xv + 191 pp.

Manuscript received May 7, 1971.

399

AN ALBINO DE KAY’S SNAKE (STORE RI A DEKAYI WRIGHTOR UM TRAPIDO) FROM CENTRAL ILLINOIS

JAMES H. THRALL

Illinois State University, Normal, Illinois 61761

Abstract. — The second record of an albino De Kay’s snake from Illinois.

An albino snake, Storeria dekayi wright- orum Trapido, collected 1.5 miles west of Lexington, McLean County, Illinois, was brought to me in June of 1969. This ani¬ mal lived until August 27, 1969, when it died from unknown causes. It was pre¬ served and placed in my personal collec¬ tion (J.H.T. 33).

Two other albino De Kay’s snakes have been reported, one from Illinois (Hensely, 1959). The first albino De Kay’s snake reported from Illinois, although not pre¬ served, is described by Smith (1961) as having been cream colored with pink dor¬ sal spots. The albino reported here is pink

with pinkish brown dorsal spots and pink eyes and tongue.

Acknowledgement I would like to thank Mrs. Barbara Gardener and family who collected the snake and were kind enough to bring it to me.

Literature Cited

Hensley, M. 1959. Albinism in North American Amphibians and Reptiles. Publications of the Museum, Michigan State University. 1:133-59.

Smith, P. 1961. The Amphibians and Reptiles of Illinois. Ill. Nat. Hist. Surv. Bull. 28. Urbana, Illinois.

Manuscript received May 20, 1971.

400

HARLOW BURGESS MILLS 1906 - 1971

ROBERT A. EVERS

State Natural History Survey, Urbana, Illinois 61801

Figure 1. H. B. Mills, March, 1963.

Harloyi Burgess Mills (Figure 1), re¬ tired Chief of the Illinois Natural History Survey, retired visiting professor of ento¬ mology at the University of Wisconsin, Racine, and a long time member of the Illinois State Academy of Science, died in Breckenridge Hospital, Austin, Texas, on April 5, 1971, at the age of 64 years, 7 months, and 16 days. Dr. Mills had suffered a stroke five days before his death.

Harlow B. Mills was born August 20, 1906, in Le Grand, Marshall County, Iowa, son of E. M. and Anna Burgess Mills. His boyhood was spent in this small, central Iowa community. After graduation from high school, he entered Iowa State Col¬ lege of Agriculture and Mechanic Arts (now Iowa State University) and received his B.S. degree in 1929 and his M.S. de¬ gree from the same institution in 1930. During the year 1929-1930, he was also an assistant in the Iowa Experiment Sta¬ tion.

On August 27, 1930, Harlow B. Mills

and Esther Winifred Brewer of Central City, Iowa, were married. They moved to Texas where he served as assistant pro¬ fessor of entomology at Texas Agricul¬ tural and Mechanical College (now Texas A. & M. University) for the year 1930- 1931. He was also associated with the Texas Experiment Station. The couple returned to Iowa in 1931 and Harlow continued his studies at Iowa State. In 1932 he was field assistant in the experi¬ ment station. He completed his Ph.D. degree in 1934. His doctoral dissertation was “A monograph of the Collembola of Iowa.”

Dr. Mills spent the next 13 years in the west. From 1934-1935 he was Ranger Naturalist and Wildlife Technician at Yellowstone National Park. In 1935, he became Assistant State Entomologist for Montana and, in 1937, he joined the faculty of Montana State College (now Montana State University), Bozeman, as professor of zoology and entomology and head of the department. The same year he received a grant from the Bache Fund for a study of Collembola. During his professorship at Montana State he pub¬ lished numerous articles on economic en¬ tomology.

On March 1, 1947, Dr. Mills took over the duties of Chief of the Illinois Natural History Survey. His broad biological back¬ ground plus his ability to get people to cooperate were instrumental in the de¬ velopment of the Survey during his ad¬ ministration. He retired from the Survey on September 30, 1966.

After retiring from the Survey, Dr. Mills became visiting professor of ento¬ mology at the University of Wisconsin, Racine, and also served as acting dean for a time. He retired from university duties in 1970, and moved to San Mar¬ cos, Texas.

Dr. Mills was active in the Illinois State Academy of Science. He had been in Illinois but two months when he spoke at the Peoria meeting, May 2, 1947, in the general session on “The Newer Con¬ cept of Predation.” He was a member of the Conservation Committee from 1947- 1963, its chairman from 1947-1949. He was also a member of the Public Rela¬ tions Committee from 1948-1957, the Re¬ search Development Committee, 1948- 1949, and the Nominating Committee,

401

402

Notes

1961-1962. He became First Vice-Presi¬ dent for 1957-1958 and President, 1958- 1959. He was a member of the Council for the year 1959-1960.

Other professional societies in which Dr. Mills was active included the Ameri¬ can Association for the Advancement of Science, the American Fisheries Society, the American Ornithologists Union, the Ecological Society of America, the En¬ tomological Society of America, and the Wildlife Society (editor of its journal, 1947-1949). He served as President of the Montana Academy of Science in 1943. He was a member of honor and professional fraternities: Gamma Sigma Alpha, Phi Mu Alpha, Phi Sigma, and Sigma Xi. His civic activities included membership on the Council of the University Y.M.C. A. and he was for many years active in Ro¬ tary.

In 1950, Dr. Mills was a Gugenheim Fellow and spent six weeks in Jamaica, Haiti and eastern Cuba in the study of the Collembola. In 1958, he was a con¬ sultant for both the U.S. Department of Agriculture and the Department of In¬ terior on the fire ant control programs in the southern states. His duties were to find the strong and weak points of the program and to effect a reconciliation be¬ tween divergent viewpoints. From Au¬ gust 1962 to August 1963, he was Chief Scientist in the Latin American Office of the National Science Foundation, Rio de Janerio, Brazil. In this position he repre¬ sented American science to the Latin American scientist, and vice versa, and sought to obtain the best possible rapport between these two groups. From January 14, 1964 to September 30, 1966, he was an advisor to the Illinois Nature Pre¬ serves Commission.

Dr. Mills had a broad knowledge of plants and animals. It was a pleasure to be in the field with him and he enjoyed being in the field with Survey staff mem¬ bers. He had a great interest in birds and, as a result of that interest, one never traveled with Harlow Mills without list¬ ing the birds observed during the trip. He knew the birds of the midwest by sight and many by their song. A visit in the woods with him was a rewarding ex¬ perience. His biological knowledge and his love for children resulted in approxi¬ mately 25 articles on scientific matters, published in children’s magazines. He en¬ joyed poetry and was, himself, a poet by avocation. His love of birds was reflected in his poems: The Nighthawk, The Wood Thrush, The Horned Lark.

Usually Dr. Mills was a happy man but tragedies strike all families. Tragedy struck Dr. Mills in June 1969, when his

beloved and devoted wife passed away. That year he published a booklet of poems, “Eleven Personal Poems of Har¬ low B. Mills,”, dedicated to the memory of Esther Brewer Mills. The love and de¬ votion of this couple are reflected in these poems. Following his wife’s death, Har¬ low went back to Wisconsin and then to Texas where his life came to a close.

Dr. Mills is survived by three children: Dr. David Mills of Silver Spring, Mary¬ land, Gary Mills of Edina, Minnesota, Mrs. Timothy (Judith Anne) Lewis of Champaign, Illinois, and 5 grandchildren. He is also survived by three brothers — Max of Marshalltown, Iowa, Ralph of Marion, Iowa, and Glenn of King Hill, Idaho — and three sisters— Mrs. Helen Grimes of Marshalltown, Iowa, Mrs. Ur¬ sula Johnson of LeGrand, Iowa, and Mrs. Marjorie Vendervelde of Emmetsburg, Iowa.

The world has lost a gentle man and a gentleman.

Technical Publications of Harlow Burgess Mills

1930

A preliminary survey of the Collembola of Iowa. Can. Entomol. 62(9) :200-203. Biological notes on aphids affecting ap¬ ples with special reference to vitality of eggs (Aphididae, Homoptera) (with D. L. Moody). J. Econ. Entomol. 23 (5) :822-825.

1931

New nearctic Collembola. Am. Museum Novitates 464. 11 p.

Notes on the oviposition of Metapterus annulipes (Stal). (Hemiptera, Reduvii- dae.) Bull. Brooklyn Entomol. Soc. 26(2) :84.

Springtails as economic insects. Proc. Iowa Acad. Sci. 37:389-392.

1932

Catalogue of the Protura. Bull. Brooklyn Entomol. Soc. 27(2) :125-130.

New and rare North American Collem¬ bola. Iowa State Coll. J. Sci. 6(3):263- 276.

The life history and thoracic develop¬ ment of Oligotoma texana (Mel.) (Em- biidina). Ann. Entomol. Soc. Am. 25 (3) :648-652.

1933

Collembola from the State of Washington (with A. R. Rolfs). Pan-Pacific En¬ tomol. 9(2) :77-83.

1934

A monograph of the Collembola of Iowa. Iowa State Coll. Div. Industr. Sci.

Notes

403

Monogr. No. 3, Collegiate Press, Inc., Ames, Iowa. 143 p.

Cotton flea hopper: varietal resistance (with F. L. Thomas and S. E. Jones). Texas Agr. Exp. Sta. Forty-sixth Ann. Rept., 1933, p. 43.

Cotton bollworm: biological studies (with F. L. Thomas). Ibid. p. 47.

1935

New Collembola from western North America. Bull. Brooklyn Entomol. Soc. 30(4) :133-141.

1936

The Virginia creeper leafhopper (with J. H. Pepper). Mont. Agr. Sta. Bull. 314, 4 p.

Observations on Yellowstone elk. J.

Mammal. 17(3) :250-253.

Heat radiation of domestic animals dur¬ ing severe cold weather. Ibid. 1 7 ^3) : 282-283.

1937

A North American Oncopodura (Collem¬ bola). Can. Entomol. 69(3):67-69.

A preliminary study of the bighorn of Yellowstone National Park. J. Mam¬ mal. 18(2) :205-212.

Observations on Grylloblatta campodei- formis Walker (with J. H. Pepper). Ann. Entomol. Soc. Am. 30(2) :269-275. Bugs, birds and blizzards in the Yellow¬ stone. Iowa State Coll. Press, Ames, Iowa. 76 p.

Some Montana birds Their relationship to insects and rodents. Mont. Agr. Exp. Sta. Circ. 151, 48 p.

1938

Control of Mormon crickets in Montana (with 0. B. Hitchcock). Mont. State Coll. Ext. Serv. Bull. 160, 19 p. Cyanide for household pests. Mont. State Coll. Ext. Serv. Circ. 90, 7 p.

Key to the grasshoppers of Montana.

Mont. Agr. Exp. Sta. Mimeo. Circ. 9. Collembola from Yucatan caves. Carne¬ gie Inst. Wash. Pub. 491:183-190. Contributions to the knowledge of the genus Sminthurides Borner (with J. W. Folsom). Bull. Museum Comp. Zool. 82(4) :229-274.

The use of tillage methods in grasshopper control (with Ralph D. Mercer). Mont. State Coll. Ext. Serv. Circ. 99, 4 p.

1939

Montana insect pests for 1937 and 1938. Twenty-seventh report of the State Entomologist of Montana. Mont. Agr. Exp. Sta. Bull. 366, 32 p.

Remarks on the geographical distribution of North American Collembola. Bull. Brooklyn Entomol. Soc. 34(3) :1 58-1 61 .

Insects in relation to soil conservation. A supplement in M. P. Hansmeier, Soil drifting on cropland in the plains area of Montana. Mont. State Coll. Ext. Serv. Bull. 176:42-46.

1940

The effects on humans of the ingestion of the confused flour beetle (with J. H. Pepper). J. Econ. Entomol. 32(,6):874- 875.

A new Nicoletia (Thysanura, Lepismati- dae) from Florida. Entomol. News 51 (10) :271-272.

1941

Montana insect pests, 1939 and 1940. Twenty-eighth report of the State En¬ tomologist. Mont. Agr. Exp. Sta. Bull. 384, 28 p.

Shelter belt insects. In E. E. Isaac, Shel¬ ter belts for Montana. Mont. State Coll. Ext. Serv. Bull. 194:17-20.

Two human diseases which may be con¬ tracted from Montana rodents. Mont. State Board Entomol. Misc. Pub. 1,

8 p.

The effect of tillage on grasshopper eggs.

J. Econ. Entomol. 34(4) :589.

Another infestation of a school building by Oeciacus vicarius Horvath (with D. J. Pletsch). Ibid. 34(4) :575.

The genus Oncopodura (Collembola).

Mont. Acad. Sci. Proc. 2:61-63. Recently introduced livestock parasites (Abstract). Ibid. 2:63.

1942

Montana insect pests, 1941 and 1942. Twenty-ninth report of the State En¬ tomologist. Mont. Agr. Exp. Sta. Bull. 408, 36 p.

Investigations in economic zoology and entomology by the Montana station. Mont. Agr. Exp. Sta. Bien. Rept. 1941- 42, 37 p.

1943

Siphonaptera - Species and host list of Montana fleas (with Wm. L. Jellison and Glen M. Kohls). Mont. State Board Entomol. Misc. Pub. 2, 22 p.

Starlings nesting in Montana. Condor 45 (5) :197.

An outbreak of the snipe fly Symphorom- yia hirta. J. Econ. Entomol. 36(5) :806.

1944

The wheat stem sawfly in Montana.

Mont. Agr. Exp. Sta. War Circ. 6, 7 p. The wheat stem sawfly {Cephas cinctus) can be controlled. Mont. Farmer 32 (5) :1, 10.

404

Notes

1945

Montana insect pests, 1943 and 1944. Thirtieth report of the State Entomolo¬ gist (with J. A. Callenbach and J. F. Reinhardt). Mont. Agr. Exp. Sta. Bull. 425, 30 p.

Some Montana birds: their relationship to insects and rodents. Mont. State Coll. Ext. Bull. 229, 48 p.

1946

Cattle grubs in Montana (with Hadleigh Marsh and Fred S. Wilson). Mont. Agr. Exp. Sta. Bull. 437, 16 p.

1947

Montana insect pests, 1945 and 1946. Thirty-first report of the State En¬ tomologist (with 0. B. Hitchcock and Ralph Schmiedeskamp). Mont. Agr. Exp. Sta. Bull. 442, 22 p.

1948

Think before you shoot. Illinois Conser¬ vation 13(1) :6-7.

Illinois State Natural History Survey Di¬ vision 1946-47, p. 86-95. In [Ill.] Dept. Registr. Educ. Ann. Rept. July 1, 1946 to June 30, 1947.

New North American Tomocerinae. Ann. Entomol. Soc. Am. 41(3) :353-359.

Fish, game, and the Illinois Natural His¬ tory Survey. Illinois Wildlife 4(1) :4-6.

1949

Natural History Survey, p. 580-587. In Blue Book State Ill. 1947-48.

1950

Natural History Survey Division, p. 145- 157. In [Ill.] Dept. Registr. Educ. Ann. Rept. July 1, 1948 - June 30, 1949.

Order Collembola, p. 91-92. In Entomo¬ logical Section, Pest Control Tech¬ nology, National Pest Control Assoc., Inc., New York.

Shot alloys and poisoning in waterfowl. Sports Afield 123(6) :122-123.

1951

Natural History Survey, p. 571-577. In Blue Book State Ill. 1949-50.

Facts and waterfowl. North Am. Wild¬ life Conf. Trans. 16:103-109.

Hunting and fishing in 2000 A.D. Out¬ door America 16(5) :4-6.

The arborist’s job. Arborist’s News 16 (10) :93-101.

Duck hunting in 2000 A.D. Wildlife in North Carolina 15(12) :9, 20.

For legislators — kill doves or not. Out¬ doors in Illinois 17(1) :3.

Illinois Natural History Survey Division, p. 141-156. In [Ill.] Dept. Registr. Educ. Ann. Rept. July 1, 1949 - June 30, 1950.

1952

Illinois Natural History Survey Division, p. 155-171. Ibid. July 1, 1950 - June

30, 1951.

Weather and climate, p. 422-429. In In¬ sects, the Yearbook of Agriculture, 1952. U. S. Dept. Agr., Washington, D. C.

Natural resources — your security: Ap¬ praisal of the 17th North American Wildlife Conference. North Am. Wild¬ life Conf. Trans. 17:539-547.

Annual Report of the State Natural His¬ tory Survey Division, 1951-52, p. 97- 133. In [Ill.] Dept. Registr. Educ. Ann. Rept. July 1, 1951 - June 30, 1952. Natural History Survey, p. 575-582. In Blue Book State Ill. 1951-52.

1953

Collembola from arctic and boreal Cana¬ da (with W. R. Richards). J. Kansas Entomol. Soc. 26(2):53-59.

Some conservation problems of the Great Lakes. Ill. Nat. Hist. Surv. Biol. Notes

31, 14 p.

The unity of animal populations. Trans. Ill. State Acad. Sci. 46:197-202.

1954

Annual report of the State Natural His¬ tory Survey Division, 1952-53, p. 53- 79. In [Ill.] Dept. Registr. Educ. Ann. Rept. July 1, 1952 - June 30, 1953.

1955

Illinois State Natural History Survey.

Entomol. News 66(6) :161-164.

Natural History Survey, p. 616-622. In Blue Book State Ill. 1953-54.

1956

Annual report of the State Natural His¬ tory Survey Division, 1953-54, p. 53- 83. In [Ill.] Dept. Registr. Educ. Ann. Rept. July 1, 1953 - June 30, 1954. Annual report of the State Natural His¬ tory Survey Division, 1954-55, p. 55- 83. Ibid., July 1, 1954 - June 30, 1955. T. R. Malthus, certified public account¬ ant. The Humanist, 1956 (4):175-183. Natural History Survey, p. 657-664. In Blue Book State Ill. 1955-56.

1957

The Coloburella-Boernerella complex (with Fred H. Schmidt). Acta Zool. Cracoviensia 2(15) :365-373.

Annual report of the State Natural His¬ tory Survey Division, 1955-56, p. 55-78. In [Ill.] Dept. Registr. Educ. Ann. Rept. July 1, 1955 - June 30, 1956.

1958

Standing room only — 2000 A.D., p. 58-67. Assoc. Midwest Fish and Game Com-

Notes

405

missioners, Proc. 25th Annual Meeting, Bismarck, N. D., July 10-12, 1958. [Non vidi].

From 1858 to 1958. A century of biologi¬ cal research. Ill. Nat. Hist. Surv. Bull. 27(2) :85-103.

Annual report of the State Natural His¬ tory Survey Division 1956-57, p. 59-82. In [Ill.] Dept. Registr. Educ. Ann. Rept., July 1, 1956 - June 30, 1957. Natural History Survey, p. 709-715. In Blue Book State Ill. 1.957-58.

1959

Annual report of the State Natural His¬ tory Survey Division, 1957-58, p. 61-88. In [Ill.] Dept. Register. Educ. Ann. Rept., July 1, 1957 - June 30, 1958. Whooping crane in the midwest (with Frank C. Bellrose). Auk 76(2) :234-235. Pest control in the modern setting. North Am. Wildlife Conf. Trans. 24:113-118. The importance of being nourished. Trans. Ill. State Acad. Sci. 51(1 & 2):3-12.

1960

Apterygota. In McGraw-Hill Encyclope¬ dia of Science and Technology. Mc¬ Graw-Hill Book Co., Inc., New York 1:495.

Collembola. Ibid. 3:289-290.

Orthoptera. Ibid. 9:416-418.

Protura. Ibid. 11:61.

How do we justify the preservation of open space? Third Ann. Metropolitan Area Planning Conf. Proc. 3:39-41. Grandfather Burgess was a forty-niner.

J. Ill. State Hist. Soc. 53(4) :404-409. Annual report of the State Natural His¬ tory Survey Division 1958-59, p. 59-84. In [Ill.] Dept. Registr. Educ. Ann. Rept., July 1, 1958 - June 30, 1959. Natural History Survey, p. 710-715. In Blue Book State Ill. 1959-60.

1961

Collembola, p. 253. In The Encyclopedia of the Biological Sciences. Reinhold Book in the Biological Sciences, Rein¬ hold Publishing Corp., New York.

The insect’s greatest competitor. Bull.

Entomol. Soc. Am. 7(3) :1 13-1 1 6. Annual report of the State Natural His¬ tory Survey Division 1959-60, p. 65-88.

In [111.] Dept. Registr. Educ. Ann. Rept., July 1, 1959 - June 30, 1960.

1962

The naturalist and the scientist. Am.

Biol. Teacher 24(2) :105-107.

State Natural History Survey Division, p. 8-22. In [Ill.] Dept. Registr. Educ. Ann. Rept., July 1, 1960 - June 30, 1961. Natural History Survey, p. 748-753. In Blue Book State Ill. 1961-62.

1964

The importance of being nourished — A review after five years. Trans. Ill. State Acad. Sci. 57(4) -.187-189.

Stephen Alfred Forbes. Syst. Zool. 13(4): 208-214.

State Natural History Survey Division, p. 8-21. In [Ill.] Dept. Registr. Educ. Ann. Rept., July 1, 1962 - June 30, 1963.

1965

The new entomologist. Proc. North Cen¬ tral Branch, Entomol. Soc. Am. 20: 32-35.

Annual report of the State Natural His¬ tory Survey Division, p. 8-22. In [Ill.] Dept. Registr. Educ. Ann. Rept., July 1, 1963 - June 30, 1964.

1966

Annual report of the State Natural His¬ tory Survey Division 1964-65, p. 7-19. Ibid., July 1, 1964 - June 30, 1965. Man’s effect on the fish and wildlife of the Illinois River (with William C. Star- rett and Frank C. Bellrose). Ill. Nat. Hist. Surv. Biol. Notes 57, 24 p.

1967

State Natural History Survey Division, p. 9-24. In [Ill.] Dept. Registr. Educ. Ann. Rept., July 1, 1965 - June 30, 1966.

Manuscript received May 25, 1971.

Errata:

In the article, “The First Record in Il¬ linois of a population of Stethaulax marmoratus (Say) (Hemiptera: Scutel- leridae) with information on Life His¬ tory.” 64:198-200, the photograph for Fig. 1 is reversed in terms of caption, i.e. read left for right.

New York Botani

:al Garden Library

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3341 4594

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Dof., J. H. 1951. The life cycle of a land snail. Conchol., 26: 21-32, 2 tables, 3 figs.

Doe, J. H., and S. H. Jones. 1951. Mineralogy of Lower Tertiary deposits. McGraw-Hill Book Co., New York, iv -f 396 pp.

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Department of Biological Sciences, Northern Illinois University, DeKalb, Illinois 60115

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