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of the

Illinois

State Academy
of Science

Volume 64
No. I
1971 _

JUN 2 19T1

NEW YORK
BOTANICAL GARDEN1

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.

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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]

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

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

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

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

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

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

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

10 22 x Intercept

10 43 x Slope

Dipole Moment

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

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]

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

Blue-

Grass

Shrubs

Soil temp. (C) .

Surface .

2 Aug .

24.6

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

% 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

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

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

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

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.

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

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

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

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

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

Station

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

13.4

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.

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

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)

mg/1

Temp.

(C)

mg/1

Temp.

(C)

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

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

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

(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 .

58.0

17.5

9.4

4.6

2.6

2.2

SC-S .

26.3

25.0

10.3

4.3

3.4

2.6

GH-S .

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

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

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

Allen, E. G. 1967. Conditions of the
Color Change of Prodigiosin. Nature
216: 929-931.

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.

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]

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

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

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.

BIO

WO BIO

BIO

WO RO

SiM

wo wo

RE WO

AE RE

BIO

AE SiM

SiM

RE WO

BIO

WO BIO

BIO

SiM WO

RE WO

BIO

WO WO

BIO

WO WO

BIO

BIO WO

BIO

810

BIO BIO
BIO

BIO

BIO WO
BIO

RO BIO

BIO

BIO BIO

BIO

BIO BIO

BIO

BIO WO

RO RO

AE AE

SiM SiM

SiM

RO SiM

BIO WO

SiM

SiM SIM

SIM

SiM

SiM WO

SiM

SiM SIM

SiM

SiM SiM

BIO

SiM

SiM SiM

BIO

RE BIO

BIO

O 200

I _ ■ I ■ i

FEET

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

Transactions Illinois Academy of Science

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Common Name

Red bud .

Hawthorn .

Mulberry .

Iron wood .

Gray dogwood .

Ha7elrmt

Bramble .

Prickly ash .

Smilav

Common elder .

Wahoo .

Common

gooseberry .

Smooth sumac .

Scientific Name

UNDERSTORY TREES

Cercis canadensis L .

Crataegus .

Morus rubra .

Ostrya virginiana (Mill.)

Koch .

SHRUBS

Cornus racemosa Lam .

Cnrvlu r am tr in ana Walt

Rubus allegheniensis Porter . .
Zanthoxylum americanum

Mill .

Rm.il ax. h ix 'hid. a Mnhl

Sambucus canadensis L .

Euonymus atropurpureus

Jacq. . . . .

Ribes missouriense Nutt .

Rhus glabra L .

Totals .

Transactions Illinois Academy of Science

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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

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

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.

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

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).

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

15° C .

400

124

111

20° C .

100

20° C .

400

295

219

181

25° C .

100

231

130

126

835

238

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

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

15° C .

400

20° C .

100

20° C .

400

25° C .

100

25° C .

400

30° C .

100

2300

202

629

30° C .

400

602

328

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

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]

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

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

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

(±)

100

5.9 x 10-5 .

(-)

(±)

18/

5.9 x 10"6 .

(-)

(±)

5.9 x 10-8 .

(±)

Alph

a-Ketoglutarate

5.9 x 10 4 .

(-)

( + )

5 . 9 x 10~5 .

(-)

( + )

5.9 x 10 6 .

(-)

( + )

5.9 x 10 7 .

(+)

( + )

(increase)

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

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

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

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,

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.

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Hiltibran — Biochemical Action of DDT

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Colvin, H. J., and A. T. Phillips. 1968.
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Dewitt, J. B. 1956. Chronic toxicity to
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Fukami, J., and C. Tomizawa. 1956. Ef¬
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Hiltibran, R. C. 1966. Hydrolysis of

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Hiltibran, R. C. 1967a. Oxidation of

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Hiltibran, R. C. 1967c. Effects of some
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Hiltibran, R. C. 1969a. The hydrolysis
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mitochondria in the presence of 2,4-
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Hiltibran, R. C. 1969b. Oxygen and
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Hiltibran, R. C. 1971. Effects of cad¬
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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]

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

—

( IW)_

_C-* (!>,,

- o . ) _

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

Gunter et al — Nitroparaffin Surface Tensions

Av.

Dev.

Dial

Read.

No.

Read.

i~n

Prob¬

able

Error

(dyne/

cm)

(dyne/

cm)

Av.

Dev.

Dial

Read.

No.

Read.

Prob¬

able

Error

(dyne/

cm)

(dyne/

cm)

Av.

Dev.

Dial

Read.

No.

Read.

Prob¬

able

Error

(dyne/

cm)

(dyne/

cm)

Mole Fraction

(RN02)

■M

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

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OOOOOi— 'OOomcmo

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

CM »—i »-h »— h CM r— H r— i — CM i-h

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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

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

r— H

r2,l

Mole Fraction

(RN02)

30°C

35°C

45 °C

30°C

35°C

45 °C

Nitromethane

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

0.1 .

—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

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.

Enthalpy

Entropy

(ergs/ cm

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

0.1

0.137

20.3

68.4

68.2

68.4

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

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

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

9.89

0.3

0.128

18.2

65.9

65.8

65.9

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

0.8

0.122

15.5

66.1

9.20

0.9

0.111

13.5

63.4

8.59

1.0

0.104

12.0

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-

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

Mole

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

0.1

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

0.4

—0.021

—2.6

3.0

—8.9

—9.0

—1.0

0.5

—0.046

—7.1

—1.3

—18.

—2.5

0.6

—0.053

—8.3

—2.7

—20.

—3.0

—2.9

—3.0

0.7

—0.053

—8.4

—3.3

—21.

—20.

—21.

—3.0

0.8

—0.044

—7.1

—3.1

—18.

—17.

—2.6

—2.5

—2.6

0.9

—0.035

—5.5

—2.8

—14.

—13.

—2.0

—1.9

1.0

0.0

Nitroethane

0.0

0.1

—0.010

—1.9

0.80

—3.3

—3.4

—0.59

—0.63

—0.60

0.2

—0.0052

—1.0

3.0

—2.3

—2.1

—2.3

—0.38

0.3

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

—0.58

—0.63

—0.59

0.5

—0.0055

—1.3

4.5

—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

0.55

0.8

0.012

1.9

5.9

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

Gunter et al — Nitroparaffin Surface Tensions

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

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

186.6

189.7

.8 .

180.4

180.3

180.5

181.0

181.5

184.0

.9 .

175.6

175.4

175.9

176.0

176.5

178.2

1.0 .

170.7

171.0

171.5

170.7

171.0

171.5

172.5

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.

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

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]

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

Temp.

(g/ ml-

(g/ml-

(g/ ml-

(g/ml-

d(conc=l)

(°C)

(g/ml)

cone)

cone2)

cone3)

cone4)

cone5)

(g/ml)

d VS. X2

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

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

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

Nitroethane

Temp.

(g/ml-

(g/ ml-

(g/ml-

(g/ ml-

d(conc=l)

(°C)

(g/ml)

cone)

cone2)

cone3)

cone4)

cone5)

(g/ml)

d VS. X2

1.5748

—0.4000

—0.1681

0.2476

—0.3825

0.1628

1.0346

1.5670

—0.5201

0.8910

—2.5015

2.4683

—0.8753

1.0294

1.5455

—0.4251

0.09944

—0.4724

0.4413

—0.1719

1.0168

d VS. W2

1.5749

—0.8377

0.5220

—0.3867

0.1983

—0.03624

1.0346

1.5660

—0.8859

1.3898

—3.4098

4.0286

—1.6599

1.0288

1.5452

— 0 . 8286

0.6076

—0.6634

0.5452

—0.1892

1.0168

d VS. V2

1.5749

—0.5471

— 0.004345

0.1398

—0.2630

0.1344

1.0347

1.5673

—0.6922

1.4641

—4.1008

4.5409

— 1 . 7504

1.0289

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-

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

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

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

0.000568

0.00427

0.000580

0.000925

0.00417

0.000921

0.000750

0.00488

0.000658

on co

N- O T-H
i on

O O O

VO VO oo

NOvO

Nforo

O O O

CO LO ON

o- o

O "f 1“H

O O O

odd

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

ciOON

o- —t o

O LO i-H

o o o
o o o

odd

dod

odd

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 •

d o •

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 •

o o •

d d •

Nitromethan

0.000967

0.00113

0.000775

0.000769

0.000774

0.000710

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 •

d d •

O O ;

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

nperature

(°C)

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.

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

Table 3. — The Per Cent Deviation.

Nitromethane

Nitroethane

Temp.

(°C)

d vs. X2

1.76

0.301

0.0698

0.0551

0.0603

1.19

0.129

0.0427

0.0437

0.0391

1.73

0.311

0.0821

0.0448

0.0437

1.38

0.307

0.323

0.300

0.294

2.57

1.20

0.163

0.0444

0.0474

0.0426

d VS. W2

1.20

0.150

0.0559

0.0549

0.0596

1.66

0.208

0.0696

0.0381

0.0414

1.22

0.176

0.0559

0.0510

0.0559

1.54

0.299

0.315

0.341

0.283

1.39

1.62

0.236

0.0706

0.0562

0.0549

d VS. V2

0.111

0.0545

0.0559

0.0600

0.0656

0.0554

0.0556

0.0564

0.0430

0.0416

0.141

0.0627

0.0516

0.0558

0.0552

0.391

0.359

0.369

0.378

0.298

0.224

0.0820

0.0782

0.0504

0.0516

0.0465

Vm vs. x2

0.0975

0.0474

0.0505

0.0546

0.0593

0.0424

0.0450

0.0410

0.0366

0.0303

0.127

0.0514

0.0542

0.0527

0.0482

0.333

0.301

0.319

0.294

0.286

0.220

0.0691

0.0714

0.0447

0.0462

0.0410

Vm vs. w2

4.96

1.24

0.294

0.0831

0.0610

2.06

0.406

0.0484

0.0312

0.0328

4.93

1.21

0.290

0.0951

0.0686

2.23

0.621

0.315

0.322

0.272

6.16

2.10

0.378

0.0703

0.0498

0.0516

Vm vs. v2

3.12

0.490

0.0855

0.0541

0.0577

0.866

0.0616

0.0361

0.0302

0.0288

3.09

0.472

0.0876

0.0561

0.0549

1.04

0.298

0.274

0.280

0.226

4.29

0.890

0.0539

0.0470

0.0482

0.0472

Transactions Illinois Academy of Science

Table 4. — The Multiple Correlation Coeffiecient.

Temp.

(°C)

Nitromethane

Nitroethane

d VS. X2

0.9763

0.9771

0.9890

0.9994

0.9993

1.000

1.0000

0.9930

0.9905

0.9927

0.9999

0.9996

0.9999

1.0000

0.9996

1.0000

0.9997

1.0000

0.9998

1.0000

d VS. W2

0.9890

0.9998

1.0000

0.9864

0.9998

1.0000

0.9886

0.9998

1.0000

0.9882

0.9996

0.9998

0.9968

1.0000

0.9868

0.9997

1.0000

d VS. V2

0.9999

1.0000

0.9998

1.0000

0.9992

0.9994

0.9995

0.9998

0.9999

1.0000

Vh vs. x2

1.0000

0.9990

0.9993

0.9995

0.9996

1.0000

Vm vs. w2

0.9371

0.9965

0.9998

1.0000

0.9600

0.9986

1.0000

0.9377

0.9966

0.9998

1.0000

0.9534

0.9968

0.9993

0.9994

0.9996

0.9758

1.0000

0.9594

0.9988

1.0000

Vm vs. v2

0.9750

0.9994

1.0000

0.9930

1.0000

0.9753

0.9995

1.0000

0.9899

0.9993

0.9995

0.9997

0.9883

1.0000

0.9927

1.0000

Musulin — Computerized Curve Fitting

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.

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

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.

Temperature

(°C)

Nitromethane

Nitroethane

(g/ml)

-ai

(g/ml-

conc)

(g/ ml-
conc2)

d(Conc

=1)

(g/ml)

-ai

(g/ ml-
conc)

(g/ ml-

cone2)

Per Cent
d(conc
-1)

d VS. X2

25a

1.5796

0.1977

—0.2447

1.1372

1.5702

0.2009

—0.2462

1.1231

1.5727

0.3755

—0.1608

1.0364

1.5619

0.2042

—0.2404

1.1173

1.5629

0.3504

—0.1829

1.0296

1.5456

0.2378

—0.2101

1.0977

1.5426

0.3637

—0.1592

1.0197

“Brown & Smith (1955)

d VS. W2

1.5723

0.6217

0.1686

1.1192

1.5709

0.7572

0.2236

1.0373

1.5636

0.6207

0.1706

1.1135

1.5628

0.7378

0.2053

1.0303

1.5456

0.6504

0.2025

1.0977

1.5407

0.7346

0.2140

1.0201

d vs. V2

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)

(ml/ g-
conc2)

-a3

(ml/ g-
conc3)

(Cone

=1)

(ml/g)

-ai

(ml/ g-
conc)

(ml/g-

conc2)

(Cone

=1)

Vm vs. x2

97.776

43.049

54.727

97.697

25.175

72.522

98.332

43 . 290

55.042

98.097

25.327

72 . 770

99.625

43.990

55.635

99.599

25 . 894

73.705

Vm vs. w2

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

97.153

66.252

24.135

55.036

97.624

32.549

7.536

72.611

97.685

66.478

24.111

55.318

98.192

33.872

8.766

73.086

99.527

72.811

28.877

55.593

99.533

33.592

7.889

73.830

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

-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.

Nitromethane

Nitroethane

Temp.

(°C)

(ml/ mole-
(mole
fraction)2)

-ai

(ml/ mole-
(mole
fraction)3)

Error

Per Cent

(ml/ mole-
(mole
fraction)2)

-ai

(ml/mole-

(mole

fraction)3)

Error

Per Cent

Two Term Equation

0.6715

0.009510

30.3

0.01983

0.3995

86.5

0.8251

—0.04203

30.2

0.1183

0.5076

109

0.08515

1.0004

55.2

Quadratic Term Only

0.6715

0.0000

28.6

0.01983

0.0000

142

0.8251

0.0000

28.5

0.1457

0.0000

144

1.0502

0.0000

0.08515

0.0000

141

Cubic Term Only

0.0000

0.009510

131

0.0000

0.3995

82.7

0.0000

—0.04203

136

0.0000

0.5378

114

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-

Transactions Illinois Academy of Science

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

-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

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

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

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

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”.

Literature Cited

Baten, W. D. 1938. Elementary Mathe¬
matical Statistics. John Wiley & Sons,
Inc., New York, New York, x + 338

pp.

Bennett, C. A. and N. L. Franklin.
1954. Statistical Analysis in Chemistry
and the Chemical Industry. John Wiley
& Sons, Inc., N.Y., xvi & 724 pp. 434-5.
Boyd, G. E. and C. E. Copeland. 1942.
Surface Tensions, Densities, and Para-
chors of the Aliphatic Nitroparaffins.
Jour. Am. Chem. Soc. 64, 2540-2543.
3 tables.

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.

1 table, 3 figures.

- . 1955. Liquid Vapour

Equilibria. VII. The Systems Nitro-
methane + Benzene and Nitromethane
+ Carbon Tetrachloride at 45°C. Aus¬
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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

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)

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

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

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

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

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.

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

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.

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

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.

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

ch3co2 Na
CH3CN

_ 00^00„ ^
<§>c!-c^>c-c^o) 4- 2 (op

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]

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

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Quoted passages, titles, and citations must be checked and rechecked
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Malcolm T. Jollie,

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

Transactions

of the

Illinois

NEW

} K?

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.

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.

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

Chromosome

Number

Distribution

C. aethiopicus Clayton et Harlan .

18, 36

East Africa; Ethiopia to Transvaal

C. arcuatus J. S. Presl ex.

C. B. Presl .

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

Indonesia to Australia

C. barberi Rang, et Tad .

South India

C. dactylon (L.) Pers

var. dactylon .

Cosmopolitan

var. afghanicus Harlan et de Wet .

18, 36

Afghanistan

var. aridus Harlan et de Wet .

South Africa to Palestine to

South India; intro, in Hawaii,
Arizona

var. coursii (A. Camus)

Harlan et de Wet .

Madagascar

var. elegans Rendle .

Southern Africa; Mozambique,
Zambia and Angola southward

var. polevansii (Stent) Harlan

et de Wet .

Baberspan, South Africa

C. incompletus Nees

var. incompletus .

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 .

Ethiopia, Uganda, Kenya,

Tanzania

C. transvaalensis Burtt-Davy .

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

Very Open

Large

None

C. arcuatus .

None

Open

Small

None

C. barberi .

None

Open

Small

None

C. dactylon

var. dactylon .

A & B

A, R, B

B, C

Dense

Small-

Medium

V ariable

var. afghanicus .

None

Very Open

Medium

Yes

var. aridus .

Open

Small-

Medium

None

var. coursii .

None

Dense

Large

None

var. elegans .

Lax

Medium

None

var. polevansii .

Dense

Small

Yes

C. incompletus

Yes

var. incompletus . . . .

None

Dense

Small

var. hirsutus .

None

Dense

Small

Yes

C. nleriifuensis

var. nlemf uensis . . . .

None

Open

Medium-

Large

None

var. robustus (lx) . . .

None

Very Open

Large

None

var. robustus (4x) . . .

None

Open

Large

None

C. plectostachyus .

None

Open

Large

None

C. transvaalensis .

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) .

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

—

15 + 4*

—

* 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.

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.

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

(Eyring, Walter and Kimble, 1947) :

k=o

m=-k

4tt

2k+l k+1

<t> )

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

= 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)

/ - 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)

— 0 ; qX3U (x)V (x) dx + r 1 QU (x)V (x) dx r2dr.

122

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 “

r .

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

lh(i)

> =

j( we

can use similar

£(1)

S(l)

foU)

£< 2)

S(2)

f„(2)

£(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)

fo(2)

^123

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)

^ (1)

^o(D

S(3)

$o(3)

S(2)

(2)

S(3)

^o(3)

^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

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

Days

Planting

(cm)

After

Planting

Length

% DC

Length

% DC

Length

% DC

Harosoy (Planted April 29)

2.5

6.6

1.7

7.0

5.3

7.7

8.5

4.0

5.0

8.1

1.9

8.2

6.2

8.0

9.2

4.4

Kent (Planted May 20)

Sub-surface

4.5

2.5

4.4

11.3

7.2

2.5

6.0

4.8

5.9

12.4

7.8

5.0

8.2

5.0

8.7

11.7

7.5

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

Planting

(cm)

Per cent of hypocotyl with dead
cells; days after planting

2.5

5.0

2.5

5.0

2.5

5.0

2.5

5.0

2.5

5.0

2.5

5.0

2.5

100

5.0

2.5

5.0

2.5

5.0

2.5

5.0

2.5

5.0

2.5

5.0

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

Planting

(cm)

Per cent ol
with dead
after p

hypocotyl
cells; days
[anting

Kent .

sub-surface

2.5

5.0

Lindarin .

sub-surface

2.5

5.0

Merit .

sub-surface

2.5

5.0

Norcheif .

sub-surface

2.5

5.0

Portage .

sub-surface

2.5

5.0

Wayne .

sub-surface

2.5

5.0

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]

148

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® .

6.0 x 102

4.4 x 103

0 .

6.6 x 105

4 x 103

1 .

5. Ox 105

4 x 104

2 .

1.0 x 105

1.0 x 103

3 .

2.0 x 105

5 x 104

4 .

1.0 x 105

2 x 105

8 .

2.0 x 104

1.5 x 106

24 .

2.0 x 104

4.0 x 104

1.8 x 105

5.0 x 104

36 .

2.0 x 104

4.3 x 105

1.0 x 104

5.0 x 102

2.5 x 103

1.0 x 103

3.0 x 103

48b .

7.0 x 103

5.0 x 105

1.0 x 103

2.0 x 10s

3.0 x 103

1.0 x 105

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 .

.5 .

1 x 104

15 .

3 x 105

5 x 104

20 .

6 x 103

3 x 102

3 x 104

30 .

7 x 104

1 x 103

2.5 x 104

5 x 103

40 .

1 x 103

2 x 103

60 .

2.1 x 103

2 x 103

70 .

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

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

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

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

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 104

9 x 103

2 x 104

3 weeks .

2 x 104

1 x 104

5 x 103

5 x 105

1 x 104

2 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

1 . 89

1— 1

i— i

. — .

1 — 1

•— 1

i — i

„ — _

■ — "

1 — "

» — i

i— i

, |

► — <

Ll_

- -

—

LlJ

Li_

1—

(—

_ 1

—i

LlJ

:=)

(_>

C_J

_ 1

1— 4

*— H

1 -

►— *

1 — *

Cl.

0.16 0.12

T7Tr-r-r

mg/1

QJ <u

OJ QJ

U_ U_

O Q

<_>

_ l

i—i

t—

1—

cr:

LT)

<S>

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

164

Transactions Illinois Academy of Science

Fe(II) CONCENTRATION, mg/liter

Wang and Evans — Behavior of Iron

165

166

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).

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.

168

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.

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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.

180

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

182

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*- -/

Conformation

solar energy

A - Conformation

Synthesis of X

C - * - Constituents

of I

Potential

energy

Substance H

Constituents

of n

Solar

energy

♦ . . Coupled .
reactions

♦

Synthesis of

Conformation I

Constituents of I

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

190

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

192

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]

194

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 .

Verschuren (1955)

Rhinolophus euryale .

Dorst (1957)

Anoura caudifera .

Linares (1967)

Glossophaga longirostris .

Setzer (1950)

Antrozous p. pallidus .

Setzer (1950)

Barbastella barbastellus .

Numerous

Palasthy (1968)

Eptesicus capensis .

Allen (1940)

Lasiurus borealis .

Allen (1940)

Hamilton-Smith (1968)

Dorst (1957)

Miniopterus schreibersi .

Myotis daubentoni .

Haensel (1968)

Myotis grisescens .

Tuttle (1961)

Myotis lucifugus .

Dubkin (1952) and this report

Myotis sodalis .

Numerous

Metzger (1956)

Barbour and Davis (1970)

Myotis velifer incautus .

Rogers (1965)

Nyctalus noctula .

Dulic and Mikuska (1968)

Nycticeius burner alis .

Easterla and Watkins (1968)

Pipistrellus pipistrellus .

Allen (1940)

Pipistrellus s. subflavus .

Blair (1948)

Mollossus fortis .

Heatwole et al. (1964)

Molossus tropidorhynchus .

Allen (1940)

Tadarida b. mexicana .

Numerous

McCoy (1960) and Glass (1954),
Herreid and Davis (1960)

Tadarida ( Chaerephon ) plicatus . .

Allen (1940)

Tadarida femorosacca .

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 .

6.7 + .2

5. 7-7. 3

4.5 + .1

3. 8-4. 9

7.6 + .1

7. 1-8.3

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

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

. R33S

o\ .

of the

Illinois

State Academy

of Science

LIBRARY

MAR, zz m

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

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

208

Transactions Illinois Academy of Science

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.

210

Transactions Illinois Academy of Science

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.

Total

200

33.5

202

9.9

198

42.9

199

31.8

197

41.6

200

102

114

57.0

200

34.8

200

22.5

200

44.5

200

100

50.0

199

36.7

Mean

200

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-

214

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

216

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

218

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.

220

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.

224

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

228

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

*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

94.1

84.7

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

16.5

14.1

11.7

9.4

7.0

4.7

2.3

0.0

+ + +
+ + +
+ + +
+ + +
+ +

A,E**

A,E

A,E,M

M,N

E,M,N

A,E,M

E,M

A,E

A,E,M

A,E

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

Occurrence

Concentrations (ppm)*
Mean Median

Highest

Kidneys

Juveniles

3/10

<0.06

0.12

Adults

4/10

0.07+ 0.04

<0.06

0.44

Liver

Juveniles

3/10

<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

0.28

Leg Muscle

Juveniles

3/10

0.02+ 0.01

<0.02

0.10

Adults

2/10

<0.02

0.08

Sternal Muscle

Juveniles

1/10

<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)

3.42

8.65

6.68

7.24

7.08

7.69

9.14

6.32

8.41

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

66.67

63.30

60.40

51.57

48.97

49.29

39.40

35.07

1.84

64.83

61.46

58.56

49.73

47.13

42.45

37.66

33.23

31.60

28.23

25.33

16.50

13.90

9.22

4.33

27.27

23.90

21.00*

12.17

9.57

4.89

22.38

19.01*

16.11

7.28

4.68

17.70*

14.33

11.43

2.60

15.10

11.73

8.83

6.27

2.90

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

Replications

NA x 10"6 cm2
Infected

Percent

Non-infected

Infected

location 10.

2.9

6.1

Infected

locations 10 through 9.

3.2

5.5

Infected

locations 10 through 7.

3.4

5.6

Infected

10, 10 through 9, and 10 through 7, above.

9.7

111

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:

County

Coal Meas

Coal Measures

Dev

Devonian

figure(s)

group

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

limestone

Miss

Mississippian

Missouri

M&W

Meek and Worthen

N&W

Newberry and Worthen

number

Ord

Ordovician

page(s)

Paleontographica

Americana

Plate

ppa

Proceedings, Philadelphia
Academy of Natural
Science

part

Sil

Silurian

Sandstone

STJ&W

St. John and Worthen

U&E

Ulrich and Everett

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.

262

Transactions Illinois Academy of Science

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

274

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).

276

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-

282

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

284

Transactions Illinois Academy of Science

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

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

288

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

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

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

294

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.

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-

296

Transactions Illinois Academy of Science

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.

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)

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

3.65 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

302

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

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Literature Cited are as shown below:

Doe, J. H. 1951. The life cycle of a land snail. Conchol., 26(3):
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Doe, J. H., and S. H. Jones. 1951. Mineralogy of Lower Tertiary
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Transactions

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Illinois

State Academy

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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
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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

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-

313

314

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

2.566

2.823

2.759

0.135

0.128

0.110

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

0.862

1.093

1.118

16.32

0.072

0.078

0.071

1.85

0.319

0.301

0.290

3.45

0.286

0.266

0.264

4.07

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.

316

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.

317

318

Transactions Illinois Academy of Science

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.

320

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.

322

Transactions Illinois Academy of Science

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

324

Transactions Illinois Academy of Science

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.

326

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).

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)

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:

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

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

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

of sandy silt and brown gravel.

La Grange

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

Till, yellow-gray-brown sandy
silt, clayey silt, and sand to
clayey sand with trace of silt.

Wheaton-Lombard-

Glen Ellyn

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

OH(MC0^i0CDt>00(JlOH(NC0T)<i0?Dt>0005OH
l— IMCO^lOlXlOOOaiHlHHr- It— 1 i— 1 i— It— It— It— ICVICV1CV1CVICV1CV1CV1CV1CV1CV1COCO

oocxdoocxjoooooocooooooooooooo oo'oo'^'oo'oo'qo'oo'oo'oo'oo'oo'oo'oo'oo'oo'oooo'

TOTAL

^uno3
1*40 J, XlJBQ

148

ioiooocoiot>ooioaicoLooooHt>i>oooHt'ixiHiooiooco

2480

l-H

iSunj -osi]^[

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105

4.23

^SU0^

03 03

OCOOOOOHHNIO<M^«Dt)ihiMOOOOOOOOIMWO

T—I r— 1

2.42

modsodid

o o

1— IOOOCVIOCVItHOOOOC—OOCOOOOOOOUOOOOO

T— 1

0.85

mninpiud fP

i-H t>

^Hoc^Hooa)io^c^ooo^(Mw^t>c<i^a)co^^t>ai

04 T—i <X> CO 04 T—I CM lO CO T—I T—| T—I CO Ml T—I r— 1 T— 1 T—I

562

22.67

AOdnp\[

Cvl O

<*<1

OOOCnOOOOOHOlOOOONOOOIMOOOOrtOO
1-1 03

2.70

uiiupudpouuojp

<M O

PhPh

^OO^OCOOOiCDcO^CO^OOiOOOCiOlH^OCOOOOlOO
04 CO 04 t—i r— 1

223

8.99

mmxods

-oipuiwidjp

t- o

T—i

'A 'A

'tOlMlMOOiCOOOOOCOOOOOt'OOOHOOOlMO

T— 1 T—I

2.54

mnuvsntf

CO 03

186

7.50

mnooooidpi

o o

T— 1

0.52

snuiBxddsy

o o

T—i

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CO t—i CO t—i , T—I 04 CO

258

10.40

vumudiiy

t- 03

T—I

922

37.18

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

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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.
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_ , 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¬
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Involvement of the Central Nervous
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Gupta, S. L., 1956, Occurrence of Asper¬
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Aspergillus Infektion Nach Operation
Eines Aorten-Stanose, Schweiz, Med.
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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¬
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De Bartolo — Mould Spores in the Atmosphere

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Seed Rot and Seedling Blight of Sor¬
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Black Mould of Garlic, Allium sativum
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_ , 1941, Species Specificity

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1942, The Incidence of Atmospheric
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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

6-48

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

Very Abundant

3 inches apart

Abundant

6 inches apart

Very Common

1 foot apart

Common

1.5 feet apart

Very Frequent

2 feet apart

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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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

17.0

Glyceria striata

F-

4.8

Carex lurida

0.4

Cardamine bulbosa

2.0

Pilea fontana

• • •

0.2

Caltha palustris

...

2.0

Solidago patula

F-

4.0

Aster laterifiorus

F-

1.6

Oxypolis rigidior

. . •

F-

1.0

Equisetum arvense

...

0.2

Cinna arundinacea

F-

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

16.5

Pilea fontana

VCL

32.3

Agrostis alba

9.6

Carex lurida

VFL

7.1

Juncus dudleyi

....

5.1

Equisetum arvense

• • • •

....

5.3

Selaginella apoda

....

4.5

Eleocharis erythropoda

....

. , . .

....

13.6

Eupatorium perfoliatum

F-

3.6

Solidago patula

2.9

Pedicularis lanceolata

F-

1.1

Caltha palustris

2.4

Aster lateriflorus

F-

....

2.0

Chelone glabra

F-

1.3

Lobelia siphilitica

F-

1.2

Scirpus atrovirens

VFL

Oxypolis rigidior

F-

Carex vulpinoidea

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

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

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)

61*

58*

Age (Years)

19.3

College Credit Earned
(Quarter Hours)

34.2

41.0

Percent with Academic Scholarship

Percent Working

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)

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

17.2

Ulmus rubra

6.4

Ptelea trifoliata

4.2

Ulmus americana

3.3

Populus grandidentata

3.1

Malus ioensis

2.9

Carya ovata

2.7

Crataegus mollis

2.0

Prunus serotina

1.8

Quercus velutina

1.6

Celtis occidentalis

1.1

Prunus americana

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

1914

1959

Ulmus americana

1924

1959

Acer negundo

1924

1963

Quercus velutina

1918

1963

Ulmus americana

1923

1963

Juglans nigra

30-35

1923-28

1963

Ulmus americana

1928

1963

Populus grandidentata

1928

1963

Quercus velutina

1928

1963

Acer negundo

25-30

1928-33

1963

Ulmus americana

25-30

1928-33

1963

Prunus serotina

1933

1963

Ulmus rubra

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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**W = west half of gully, on upper slope
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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

244

Juglans nigra

113

Celtis occidentalis

Quercus velutina

Acer negundo

Prunus serotina

Carya ovata

Ulmus americana

Populus grandidentata

Malus ioensis

Prunus virginiana

Juniperus virginiana

Crataegus mollis

Fraxinus americana

Acer saccharum

Quercus macrocarpa

Quercus alba

Prunus americana

Ptelea trifoliata

Viburnum lentago

Catalpa speciosa

1088

Total Species Present

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

3 5185 Ol

3341 4594

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:

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.

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
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(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.

Malcolm T. Jollie,

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

Xa/td o^Xlmo&v

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