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Volume 24 K42X 1963 Number 1-2 


Taxes SSAC TIONS 
of the KENTUCKY 
ACADEMY ot SCIENCE 


Official Organ 
KEenTuCKY ACADEMY OF SCIENCE 


CONTENTS 


A Study of Homing in the Indiana Bat, Myotis sodalis 
Marion D. HassEL. 


Comparative Behavioral Characteristics of Six Genera of Mice 
Cares L. Riepy and Micwaen J. Harvey 


The Role of Geology in Conservation in Kentucky 
Preston McGrain 


Relationship Between Cholesterol and Lecithin Intake to 
Levels of Lechithin and Free Cholestrol in the 
Blood Serum of Rats 


Faye J. Bowman, Wm. G. Downs, Jr., and 
Rosert L. SUBLETT 


On the Activity of Plethodon oe as Influenced by Light 
Rocer W. BaRBouR 
An Annotated Checklist of Fishes from Dix River and 
Tributaries (Exclusive of Herrington Reservoir ) 
Puiu FE. GREESON 
Aging Bats in Winter 
Wayne H. Davis 


The Kentucky Academy of Science 
Founded May 8, 1914 


OFFICERS 1962-63 


President: Lyte Dawson, University of Kentucky 

President-elect: RicHARD CHAPMAN, University of Kentucky 

Vice President: Dwicut LinpsAy, Georgetown College 

Secretary: Gernnir Levey, Berea College 

Treasurer: J. H. B. Garner, University of Kentucky 

Representative to AAAS Council: Mary Wuanton, Georgetown College 


Counselors to Junior Academy: Maurice CuHristoPpHER, Murray State College, and THomas A. 
Hutto, Eastern State College 


OFFICERS OF SECTIONS 


BOTANY 

Chairman: Cart E. HEnniIcKson, University of Kentucky 

Secretary: RosertT LARANCE, Eastern Kentucky State College 
CHEMISTRY 

Chairman: THoMAs KanGt, Ursuline College 

Secretary: Ex.xis Brown, University of Kentucky 
GEOLOGY 

Chairman: James E. Conxrn, University of Louisville 

Secretary: Joun C. Purixey, Morehead State College 
MICROBIOLOGY 

Chairman: Emi. Kotrcnuer, University of Louisville 

Secretary: Lucia ANDERSON, Western Kentucky State College 
PHYSICS 

Chairman: Richard Hanav, University of Kentucky 

Secretary: Oris K. WouFe, Centre College 
PSYCHOLOGY 

Chairman: Joan LEE, University of Kentucky 

Secretary: Mary ELLEN Curtin, University of Kentucky 


ZOOLOGY 
Chairman: RocEer W. Barsour, University of Kentucky 
Secretary: J. G. RopricuEz, University of Kentucky 


BOARD OF DIRECTORS 


HAzEr (NOLTAU) Bie cckeseaseccocetussnerease to 1963 Wiainrram) Ge Reape iolekccee tee aees to 1965 
WALETAM, (CAT. dalcevcecoscscasesccecastseees to 1963 Rey ELS.) WALLEY: (coos thevaatdceceuncchacceencetaeeen to 1965 
Gane SANGER ee a een cscueet cones penantsuseas to 1964 FHERBERT SHADOWEN  .eccccccccercscecceseees to 1966 
Wie) Depa AYA cw sr Peek eS Le EET to 1964 Oats | WOLRE ccs iecscdscocanetoanenubens to 1966 


EDITORIAL STAFF 


Editor: RoGeR W. Barsour, University of Kentucky, Lexington, Ky. 


Associate Editors: 
(Bacteriology and Medical Technology) SzrH Grm_KERsoNn, Berea College, Berea. 
(Botany) Mary E. WHarton, Georgetown College. 
(Chemistry) Warp Sumpter, Western State College, Bowling Green. 
(Geology) Barnspara M. Conxrn, Louisville 
(Zoology) Joun M. CarPENTER, University of Kentucky, Lexington 


Membership in the Kentucky Academy of Science is open to interested persons upon nomi- 
nation, payments of dues, and election. Application forms for membership may be obtained 
from the Secretary. The TRANSACTIONS are sent free to all members in good standing. 

: Subscription rates for non-members are: domestic, $3.50 per volume; foreign, $4.00 per 
volume. 

The TRANSACTIONS are issued semi-annually. Four numbers comprise a yolume. 

Correspondence concerning memberships or subscriptions should be addressed to the 
Secretary. Exchanges and correspondence relating to exchanges should be addressed, The 
Librarian, University of Louisville, who is the exchange agent for the Academy. Manuscripts 
and other material for publication should be addressed to the Editor. 


A STUDY OF HOMING IN THE INDIANA BAT Myotis sodalis 


MARION D. HASSELL 
Department of Zoology, University of Kentucky 


Introduction 


Homing occurs in many species of bats. Griffin (1940), Davis and 
Hall (1952), Smith and Hale (1953) and Hitchcock and Reynolds 
(1942) have reported homing in Myotis sodalis, Eptesicus fuscus and 
Myotis lucifugus respectively. 

It was felt that if large numbers of M. sodalis were released at vary- 
ing distances from the point of capture more information concerning 
homing in this species might be obtained. 


Acknowledgments 


For their advice and encouragements I am indebted to Dr. Wayne 
H. Davis and Dr. Roger W. Barbour. Mr. Michael Harvey and Mr. 
Charles M. Rippy accompanied me on trips to Bat Cave. Special recog- 
nition is due Mr. Stewart Peck for his assistance in collecting and dis- 
tributing the bats. A portion of the transportation was furnished by 
the Department of Zoology of the University of Kentucky. 


Materials and Methods 

The bats were obtained from Bat Cave at Carter Caves State Park, 
Carter County, Kentucky. They were selected from the wintering 
population of approximately 100,000 Myotis sodalis. Seven hundred 
were banded with number 2 aluminum bird bands supplied by the 
United States Fish and Wildlife Service. The bands were painted red 
with dilute fingernail polish. This was to aid in distinguishing the bats 
used in this study from the many banded bats already in the cave. 

All of the bats were released within eleven hours of the time they 
were collected. They were released at twelve locations, each twelve air 
miles farther west from Bat Cave than the preceding. Although it was 
desirable to keep all release sites on a single radius, the convenience 
of highway release sites necessitated deviations off the radius. 

Larger numbers of bats were released as the distance from Bat 
Cave increased (see Table I). The first were released at 6:00 p.m. 
October 20, 1962; the last were released at 3:45 a.m. the following day. 
Approximately 45 minutes elapsed between each release point. The 
bats were banded immediately prior to their release, and at this time 
sex, band number, release site and time of release were recorded. 

The first return trip to Bat Cave was made on the weekend of 
October 26-27. At this time an attempt was made to observe all bats 


bo 


Marion D. Hassell 


Table |— Number of Myotis sodalis Released 


Release Miles from Number released 

site Bat Cave Males Females Total 
1 12 10 15 25 
2 24 y) 16 25 
3 36 10 15 25 
4 48 Wal 14 25 
5 60 Zit 24 Bit 
6 72 24 25 49 
7 84 24 26 50 
8 96 38 37 15 
9 108 21 57 lad 

10 120 39 59 98 

att SZ 29 71 100 


12 144 39 61 100 


Total 281 419 700 


in the cave. Band number and date of recapture were recorded for all 
red-banded bats observed. Subsequent trips either by the author or 
Dr. Wayne Davis were made to Bat Cave throughout the winter. 


Results and Discussion 


Fifty-six red-banded Myotis sodalis were found in Bat Cave during 
the first return trip (see Table II). Of the 56 bats recaptured 25 were 
males. Thus 8.9% of the males and 7.4% of the females had returned 
within six nights. Those bats released at the eleventh station were re- 
leased in the early morning hours of October 21st. Therefore, the one 
recaptured from this point had completed the return trip in a maximum 
of five nights. 

To date 119 or 17% of the red-banded bats have been recaptured 
at Bat Cave. This should not imply that more have not returned. Only 
48 of the 119 bats were found in the cave more than once. One can 
thus assume that there were bats which returned to the caves but were 
not observed. 

As can be seen in Table II, the total number of recaptured M. 
sodalis gradually decreased as the distance from Bat Cave increased. 
Although the percentages shown represent the present tally, approxi- 
mately the same relationship existed after each of the first three return 
trips to the cave. The decrease in percentage of return may possibly 
be accounted for by several factors. First, as Hall (1962) suggests, 
M. sodalis may return to the caves by using familiar landmarks. If 


Study of Homing in the Indiana Bat 3 


such is the case, it may be assumed that those bats which did not return 
were removed from their normal range and could not find familiar 
landmarks upon which to orinet. Secondly, those bats released west 
of Lexington were just as close to suitable caves in central Kentucky 
and southern Indiana, and they may have gonue to these. Finally, 
those which did not return were unable to sustain flight of that length 
due either to lack of energy or adverse weather conditions. 

There was considerable difference in the return percentages of 
males and females from several release points. However, the total 
returns from these points were in normal relationship to the others. 
No significant difference was found to exist in the percentage return of 
males and females when all recaptures were considered. 

One of the red-banded bats was found near Somerset, Kentucky, 
on December 13, 1962. The bat was found dead in a non-heated room 
of a house. There is no indication as to how long the bat may have 
been there. Somerset is approximately mid-way between Bat Cave and 
Louisville, the release point of this bat. However, it is about 100 miles 
south of a direct line between the two. 

No additional information has been gathered at this time to help 
determine the fate of the remaining bats which did not return to Bat 
Cave. It is hoped that additional band numbers will be reported to the 
Fish and Wildlife Service by other workers or interested parties. Until 
this happens or until other suitable caves are personally visited, the 
only avenue open is speculation. 


Table Il— Number and Percentage of Myotis sodalis Recaptured 
Release Number recaptured Percentage recaptured 
site Oct 26=2 shotall Males Females Total. 
1 ll 17 80.0 60.0 68.0 
2 a 16 8 Bo 2 64. 0 
3 rt 14 TORO. Bos BO60 
as 6 9 SO SeaMln SGNO 
5 tl 16 259 SI RANS WU iwoyd aeeg 
6 6 8 faq,0) 8.0 Og 8 
rs 3 9 5) 0) ils 18.0 
8 3 9 eS Me 1250 
9) 5) 6 Qo 3 US 7.8 
10 0 5 Be Se A 35 db 
that 1 6 3.4 7.0 Gad 
i, 0 + Be a Soil 4,0 


Total 56 i) 


4 Marion D. Hassell 


Summary 


A study of homing in Myotis sodalis has been conducted. Homing 
ability was found to decrease as the distance from the capture point 
increased. No significant difference was found to exist in the homing 
ability of males and females. 


Literature Cited 

Griffin, D. R. 1940. Migration of New England Cave Bats. Bul. Mus. Comp. 
Zool., Harvard Univ., 86: 216-247. 

Hall, J. S. 1962. A Life History of Taxonomic Study of the Indiana Bat Myotis 
sodalis. Reading Public Mus. Scientific Publ. No. 12. 

Hall, J. S. and W. H. Davis. 1958. A Record of Homing in the Big Brown Bat. 
Jour. Mamm., 39(2): 292. 

Hitchcock, H. B. and K. Reynolds. 1942. Homing Experiments With the Little 
Brown Bat Myotis 1. lucifugus (Le Conte). Jour. Mamm., 23: 15-23. 

Smith, E. and K. Hale. 1953. A Homing Record in the Bat Myotis 1. lucifugus. 
Jour. Mamm., 34: 122. 


COMPARATIVE BEHAVIORAL CHARACTERISTICS OF SIX 
GENERA OF MICE 


CHARLES L. RIPPY and MICHAEL J. HARVEY 
Department of Zoology, University of Kentucky 


There have been various studies of behavioral relationships be- 
tween different genera, species, and subspecies of mice. Most were con- 
cerned with a comparison of only two taxa. King (1957: 355) studied 
intra—and interspecific conflict of Mus and Peromyscus. Catlett and 
Shellhammer (1962: 133) compared the behavioral and _ biological 
characteristics of Mus and Reithrodontomys. Wirtz and Pearsen 
(1960: 131) observed aggressive behavior in Microtus pennsylvanicus 
and Peromyscus leucopus. Getz (1962: 351) observed aggressive be- 
havior in Microtus pennsylvanicus and Microtus ochrogaster. Foster 
(1959: 496) studied behavioral differences in Peromyscus maniculatus 
bairdi and Peromyscus maniculatus gracilis. 

This study compared behavior of six genera of mice when caged to- 
gether. 

Twenty adult and sub-adult mice representing six genera and six 
species were used in the experiment. Four Peromyscus leucopus leu- 
copus (2 males; 2 females) and three Peromyscus leucopus novebor- 
acensis (1 male; 2 females), one golden mouse, Ochrotomys nuttali 
aureolis (male), two house mice, Mus musculus ssp. (1 male; 1 
female), six prairie voles, Microtus ochrogaster ochrogaster (4 males; 
2 females), three pine voles, Pitymys pinetorum auricularis (2 males; 
1 female), and one southern bog lemming, Synaptomys cooperi 
kentucki (female). All of these mice were captured in the wild with 
the exception of the house mice which were reared in captivity. 

The golden mouse was captured two miles south of Beulah, Hick- 
man Co., Kentucky. The prairie voles, pine voles, bog lemming, and 
four of the white-footed mice (P. 1 leucopus) were captured five miles 
south of Lexington, Fayette Co., Kentucky. Two of the white-footed 
mice (P. 1. noveboracensis) were taken at Indianapolis, Marion Co., 
Indiana. One white-footed mouse (P. 1 noveboracensis) was taken two 
miles north of Paris, Edgar Co., Illinois. This particular combination 
of mice was used because of availability. 

The mice were confined in a cage 58 inches long, 24 inches wide, 
and 14 inches high, having four glass sides and a wooden floor. Com- 
mercial laboratory litter covered the floor. Food consisted of various 
seeds, rolled oats and greens. No feeding device was used. Food was 
placed on the floor of the cage. Water was furnished by a single 


6 Charles L. Rippy and Michael J. Harvey 


bottle fitted with a one hole stopper and a length of glass tubing. 
This was suspended in one corner of the cage. 

Observations were made of interspecific and individual behavioral 
patterns both when food and water were readily available and when 
food and water supplies were replaced after 24 hour periods of de- 
privation. Ten daytime and ten nighttime observations were made 
during a period of 60 days. Each observation lasted from 30 to 90 
minutes. 

When observations were made at night, illumination was fur- 
nished by a single 25 watt red bulb suspended above the cage. Finley 
(1959: 591) used ordinary red light to observe wood rats (Neotoma 
floridana) at night and found that red light, for all practical purposes, 
closely simulated nighttime conditions. This proved to be a useful 
device. The mice were much more active under red light than under 
ordinary lighting. During the daylight hours sunlight provided 
illumination in the laboratory. Little activity was noted during the 
daytime. 

Prior to their introduction into the experimental cage the mice 
were kept in smaller cages and separated by genus. All of the animals, 
with the exception of the two house mice, were placed in the cage 
at the same time. The house mice were added eight days later. 

Initial behavior consisted of random wandering and exploration, 
sniffing unfamiliar individuals, and a few brief tussles. The prairie 
voles, pine voles, and bog lemming displayed a higher degree of 
aggressiveness than the white-footed and golden mice. 

Brief fighting occurred between pine voles and prairie voles; pine 
voles and the bog lemming; and prairie voles and the bog lemming. 
On one occasion a prairie vole bit the nose of a pine vole drawing 
blood. Immediately thereafter the injured animal ran along the side 
of the cage, smearing blood on the glass wall. When prairie voles 
encountered this blood during their explorations they licked it from 
the glass. 

Pine voles were also observed fighting with one another as were 
prairie voles, although these same mice were compatible while caged 
together prior to their introduction into the larger cage. 

When next observed, after having been confined together for 24 
hours, all of the mice except the bog lemming were clumped together 
in one corner of the cage. Throughout the study, this clumping be- 
havior was observed during periods of inactivity. 

Although apparently socially accepted, the bog lemming usually 
isolated itself in a separate corner of the cage and rarely was it ob- 
served in a large aggregation. 

When the two house mice were introduced into the cage they 


Behavioral Characteristics of Six Genera of Mice i 


showed the same initial behavior as did the others: random wander- 
ing, exploration, and sniffing unfamiliar individuals. The house mice 
were docile, but some of the other mice, particularly the pine voles 
and prairie voles, displayed aggressiveness by snipping at the house 
mice. In one instance a pine vole grasped a house mouse by the base 
of the tail and dragged it across the cage. After being in the cage for 
a few hours, however, the house mice joined the aggregation. 

Half of the nighttime observations were devoted to food and water 
deprivation experiments. In these experiments the mice were de- 
prived of food and water for 24 hour periods. Food and water were 
then replaced and behavioral responses were observed. Little conflict 
over food occurred when there was a sufficient amount available for 
each mouse. When an insufficient amount of food was added, two or 
three grains of corn for example, much fighting occurred. Many 
squeaks and guttural noises were emitted during these conflicts. 
Mice with corn were pursued by deprived individuals who attempted 
to obtain the food for themselves. Fighting also occurred when water 
was replaced after periods of deprivation, especially since only one 
individual at a time could drink from the glass tubing. This fighting, 
accompanied by squeaks and guttural noises, consisted primarily of 
pushing in which each individual tried to gain access to the water. 
Several mice were usually involved. The pine voles and prairie voles 
displayed more aggressiveness and dominated the water bottle. The 
house mice, white-footed mice, and golden mouse attempted to drink, 
but were usually pushed away and forced to wait until the pine voles 
and prairie voles had finished. Occasionally the golden mouse would 
climb onto the glass tubing and hang on, head downward, by grasp- 
ing the tubing with feet and tail. In this manner it could secure water 
at intervals when other mice were fighting to drink. On these occa- 
sions, and at other times, when climbing on the wire support of the 
water bottle, the golden mouse exhibited apparent prehensility of 
the tail. The bog lemming did not attempt to drink until the other 
mice had finished. 


Summary 
1. When caged together, the prairie voles, pine voles, and bog 


lemming were more aggressive than the white-footed mice, house 
mice, and the golden mouse. 


2. Initial aggressiveness lasted less than 24 hours. 

3. The bog lemming, although apparently socially accepted, 
usually isolated itself from the other mice. 

4. All other mice exhibited a clumping behavior during periods 
of inactivity. 


8 Charles L. Rippy and Michael J. Harvey 


5. Little conflict over food occurred except when there was an 
insufficient amount available. 

6. Much fighting occurred when water was replaced after periods 
of deprivation, especially since only one individual at a time could 
drink. 

7. The golden mouse exhibited apparent prehensility of the tail 
when climbing. 


LITERATURE CITED 


Catlett, R. H. and H. S. Shellhammer. 1962. Comparison of behavioral and bio- 
logical characteristics of house mice and harvest mice. J. Mamm., 43(2): 
138-148. 

Finley, R. B., Jr. 1959. Observation of nocturnal animals by red light. J. Mamm., 
40(4): 591-594. 

Foster, D. D. 1959. Differences in behavior and temperment between two races 
of the deer mouse. J. Mamm., 40(4): 496-513. 

Getz, L. L. 1962. Aggressive behavior of the meadow and prairie voles. J. Mamm., 
43(3): 351-358. 

King, J. A. 1957. Intra- and interspecific conflict of Mus and Peromyscus. 
Ecology, 38(2): 355-357. 

Wirtz, W. O. and P. G. Pearsen. 1960. A preliminary analysis of habitat orienta- 
tion in Microtus and Peromyscus. Am. Mid. Nat., 63(1): 131-142. 


THE ROLE OF GEOLOGY IN CONSERVATION IN KENTUCKY 


PRESTON McGRAIN 


Assistant State Geologist, Kentucky Geological Survey 
University of Kentucky, Lexington, Kentucky 


Who of you has ever looked around his home, office, or car, and 
considered the importance of the Earth’s mineral resources in your 
daily affairs? What sort of living standard would we have if these 
raw materials were not exploited? How would we operate our busi- 
nesses? Clays and shales provide the raw materials for brick and 
structural tile; gypsum and limestone are used in the manufacture 
of plaster and cement; pure sands and sandstones are the source of 
silica for light bulbs and tubes, window glass, and mirrors; iron and 
aluminum ores are processed for structural steel, metal trim, plumb- 
ing, hardware, heating appliances, automobile bodies and motors, 
and cooking utensils; copper is an essential component of any elec- 
trical system; most of the electricity consumed in this area comes from 
coal-burning generating plants; and natural gas and petroleum pro- 
vide heat for buildings and homes, and fuel and lubricants for trans- 
portation. As impressive as this list might appear to be, we still 
haven't touched on such fields as ceramics, food containers, plastics, 
chemicals, fertilizers, aggregates, structural stone, mineral fibers, 
mineral pigments, mineral fillers, alloys, gem stones, precious metals, 
and numerous others. A list would range from asbestos to zinc, or 
literally from “A” to “Z.” 

These are part of the Earth’s abundant natural resources. Yet in 
one very significant aspect they differ from other natural resources— 
they are one-crop resources. Rains replenish water supplies in lakes, 
streams, and underground reservoirs. New forests will grow where 
timber crops have been harvested and, with time, soils will naturally 
regain a certain degree of fertility. But when a barrel of oil is pumped 
from the ground or a ton of iron ore is scooped from a mountain side, 
that portion of the particular mineral resource has been depleted. 

An adequate, dependable, and continuing supply of raw materials 
is indispensable to the United States and its industries in meeting the 
needs of an expanding population, a rising standard of living, and 
national defense. The demand upon our mineral resources is heavy 
and can be expected to increase in the future. Geologists and en- 
gineers are charged with the responsibility of their exploration and 
development. Although the exhaustion of mineral supplies is not im- 
minent, man’s quest for adequate food and clothing and a wide range 


10 Preston McGrain 


of material comforts, cultural advantages, education, and recreation 
are producing constantly increasing demands. 

Since mineral resources are “one-crop’ or non-renewable re- 
sources, the geologist’s concept of conservation frequently deviates 
from the orthodox definition which includes preserving, guarding, or 
keeping in a safe place. There is probably no safer place for keeping 
our petroleum and natura! gas resources than in the porous reservoir 
rocks thousands of feet underground, but can one visualize our civi- 
lization if the supply of our mineral fuels and lucricants were cut off? 
The writer's concept of the conservation of mineral resources includes 
wise and proper use, and development along constructive lines incor- 
porating economical and efficient use of mineral supplies. 

Voskuil (1955) has suggested steps necessary to maintain a steady 
flow of minerals, two of which directly concern the conservationist 
and the third, although not augmenting or conserving the original 
resource itself, has a saving consequence by increasing the efficiency 
of resource utilization. His suggestions are: 

J. The reduction of loss and waste in present mining practices. 

2. A program of ore discovery, including discovery and measure- 
ment of “sub-ore.” 

3. A program of technological improvement thru research for 
greater efficiency of resource utilization. 

Kentucky is rich in a variety of mineral resources. While it is gen- 
erally not considered a mineral state, an income of approximately 
400 million dollars annually from mineral raw materials is evidence 
of the importance on the economy of the Commonwealth. According 
to the U.S. Bureau of Mines (1961), Kentucky, in 1960, ranked 15th 
in the United States in value of minerals produced, exceeding such 
states as Alaska, Colorado, Montana, New York, and all of our border- 
ing states except West Virginia and Illinois; among the fifty states, 
Kentucky ranked second in ball clay, bituminous coal, and fluorspar 
production. (Available figures for 1961 indicate that Kentucky ranks 
third in bituminous coal production.) Only two states east of the 
Mississippi River (Illinois and Mississippi) produced more oil (U.S. 
Bureau of Mines, 1961). 

Changing processes and new industries have created new and 
varied demands. Within our generation we have seen principal coal 
markets change from fuel consumption by home and railroad to 
electric generating plants. For maximum recovery, economy, and _ use, 
surface and underground coal-mining methods have become highly 
mechanized. The same has applied to the clay industry. Yet these 
very attempts to reduce loss and waste have caused these industries 
to be targets for criticism by some conservationists, 


The Role of Geology in Conservation in Kentucky ILL 


Research and technology will increase our estimates of ultimate 
resources beyond the rather limited concept of short-term reserves. 
An example of conservation through technological research is strik- 
ingly illustrated in the recovery of petroleum by new and improved 
techniques resulting from geological and engineering studies. The 
injection of fluids to supplement natural sources of energy in the res- 
ervoir rock results in a 50- to 100-percent increase in oil recovery 
from fields using such methods. Several million barrels of oil are 
produced in Kentucky each year by water-flooding, much of this 
from fields that would have been abandoned had not secondary re- 
covery procedures been applied. Studies of such pertinent data as 
down-hole records, permeability and porosity tests, production figures, 
and subsurface geologic maps aid in determining the method to be 
used in securing the greatest ultimate recovery. The Kentucky 
Geological Survey has in its files, and available for reference by inter- 
ested parties, more than 70,000 logs and 6,500 sets of rock cuttings 
from wells drilled for oil and gas in the Commonwealth. These ma- 
terials are referred to daily by those interested in developing known 
gas and oil reserves and those searching for new production. 

Attempts at crude oil recovery from Kentucky’s natural rock 
asphalt deposits by new and improved technological research is being 
watched with considerable interest. Success here could rejuvenate 
a currently economically depressed region. 

Kentucky's statewide areal geologic mapping program, conducted 
cooperatively by the State and Federal governments through the 
Kentucky Geological Survey and the United States Geological Sur- 
vey, is providing essential knowledge for locating and developing 
our mineral wealth. The project, if carried to completion 8 to 10 
years hence, will make Kentucky a national leader in this field. Min- 
eral resource companies and exploration geologists, relating known 
deposits of asphalt, clay, coal, fluorspar, limestone, natural gas, and 
petroleum to the character and structure of the rocks in which they 
occur, can develop sound exploration programs for hidden new min- 
erals in areas where favorable rocks and structures have been mapped. 
Little-known industrial minerals such as vein deposits, evaporites, 
absorbent and catalyst clays, and high-silica sands may be disclosed 
also. These maps will create new interest in mineral exploration in 
Kentucky which, in turn, will result in new mineral wealth and in- 
dustrial development. 

Water, although not generally considered a mineral by the lay- 
man, is as dependent upon geological factors as are petroleum and 
other hydrocarbons. Adequacy of supply, replenishment of under- 
ground reservoirs, and water quality are all related to the geological 


12 Preston McGrain 


environment. A program of water conservation should include a knowl- 
edge of the local geological relationships. 

Geologic maps also provide information which enables engineers 
to more effectively select sites for the construction of dams, and plan 
flood-control and navigation structures, river development, water- 
supply and sewage facilities, and other pipeline networks. Geologic 
maps are the basic tool in the preparation of water availability maps. 
Cases could be cited where ground water investigations were delayed 
until adequate geologic maps could be assembled. And there is a 
growing trend in the use of geologic data in urban and industrial 
planning. 

Since soil classifications are in large part based on the character 
of rock formations at or near the surface of the ground, foresters, 
soil scientists, and agriculturalists obtain valuable information from 
geologic maps in planning reforestation, land utilization, and _ soil 
conservation programs. The ability of a farm pond to retain water 
is due in a large measure to the underlying soil and rock conditions. 

Conservation of our mineral resources can be accomplished by an 
orderly development and maximum recovery of our mineral commodi- 
ties. Cooperation of all parties is requisite or regulatory agencies for 
enforcing conservation practices will be established. 

Time will bring changes in our knowledge of the amounts and 
nature of the nonrenewable resources, and the grade of material 
that can be worked profitably. Dr. Thomas B. Nolan, Director of the 
U. S. Geological Survey, has recently pointed out (1962) that changes 
in adequate grade result not only from improvement in technology 
but also from the geologic accomplishment of establishing adequate 
and economic deposits of hitherto unutilized substances. 

Although the fields dealing with renewable resources receive 
more publicity than those concerned with nonrenewable resources, 
they are equally important. Geologists can assist in our State and 
National planning in both areas, and aid our people to enjoy an in- 
creasingly higher standard of living and in building a better Kentucky. 


Literature Cited 
Nolan, T. B., 1962, Role of the geologist in the national economy: Geol. Soc. 
Amer. Bull. vol. 73, no. 8, p. 273-278. 


U. S. Bureau of Mines, 1961, Minerals Yearbook 1960: U. S. Dept. Interior, 
Bureau of Mines, Minerals Yearbook, vols. 1, 2, 3. 


Voskuil, W. H., 1955, Fundamentals of mineral conservation: Jour. Geog., vol. 
LIV, no. 1, p. 34-40. 


RELATIONSHIP BETWEEN CHOLESTEROL AND LECITHIN INTAKE 
TO LEVELS OF LECITHIN AND FREE CHOLESTEROL IN THE 
BLOOD SERUM OF RATS* 


FAYE JOHNSON BOWMAN**, WM. G. DOWNS, JR., and ROBERT L. SUBLETT 


Biology Research Center and the Department of Biology and Chemistry 
Tennessee Polytechnic Institute, Cookeville, Tennessee 


It is only recently that the qualitative, as well as the quantitative, 
aspects of fat ingestion have been appreciated. The practical import- 
ance of these with regard to atherosclerosis is not clear as yet. How- 
ever, in experimental animals the formation of arterial plaques has 
been shown to be affected by the fat type as well as its abundance. 
It has been found that in association with these plaques is a high 
concentration of serum cholesterol, and in association with the high 
serum cholesterol level is a diet high in saturated fats. It has also 
been postulated that atherosclerosis results from the chylomicrons 
which follow meals. (Gofman, et. al., 1950) 

The present study had more than one objective. Since it was a 
controversial issue as to whether rats show a response to increased 
lipid intake, as other experimental animals have been found to do, 
one of the objectives was to see what response, if any, these animals 
would show when two lipids, cholesterol and lecithin, were added 
to their diet. 

Before these objectives could be fulfilled, the experimental tech- 
niques of previous investigators had to be analyzed. From these in- 
vestigations the experimental approach for this study was developed, 
with the aim of making it as closely correlated with previous studies 
as possible. 

Studies of atherosclerosis in various parts of the world, on humans 
and experimental animals, have revealed that climate, locale, strain 
and racial characteristics are not the greatest factors producing ath- 
eromatous lesions, but diet is now believed to be one of the most 
important etiologic factors. (Kreglow, 1958) 

The quantitative aspect of dietary fat has become of prime im- 
portance in the past few years, since the possible pathogenicity of 
an excessive amount of fat in the body was recognized. Some 
rather interesting and pertinent observations have been made in the 


* Aided by a grant from the Justin Potter Memorial Fund, Nashville, Ten- 
nessee. 

** Present address: Dept. of Pharmacology, Vanderbilt University, School of 
Medicine, Nashville, Tenn., U.S. A. 


14 Faye Johnson, Wm. G. Downs, Jr., and Robert L. Sublett 


past. First, it has been noted that not all animals are susceptible to 
hypercholesterolemic response after overloading the diet with choles- 
terol, and second, in animals which are susceptible it was observed 
that after a period of time they seemed to increase their capacity to 
handle this load. One may suspect on the basis of these observations 
that there exists in the animal’s organism a homeostatic mechanism 
which controls cholesterol metabolism and regulates blood choles- 
terol levels. This mechanism apparently can be stimulated under 
forced conditions of dietary overload. (Boyd, 1952) It has also been 
observed that serum cholesterol levels are seemingly influenced by 
alteration in the type of lipid intake. 

Since these observations were made much work has been done 
in connection with overloadnig the diet with a particular fat and 
observing the levels of this, and also other fats in the blood and tissues 
of the animal. 

Patil and Majar (1959) found that in tissues and serum, the level 
of phospholipids was significantly lowered in cholesterol-fed rats. 
The same investigators also found that the ratio of cholesterol to 
phospholipids was higher in cholesterol-fed groups, and that the 
cholesterol and phospholipid contents of tissues and serum of animals 
from the control groups did not show wide variations. In disagree- 
ment with this, others (Schwenk, et. al., 1959) found that the phos- 
pholipid values and the ratio of serum cholesterol to phospholipid was 
closely parallel to serum cholesterol levels, for rats receiving daily 
an increased intake of cholesterol. The serum cholesterol value was 
observed to reach a peak in 9-11 weeks and then decline, even when 
daily intake was continued, which would support the hypothesis of 
a homeostatic mechanism being in operation for the control of chol- 
esterol level in the blood. The same investigators found similar re- 
sponse in another study in which it was noted that serum cholesterol 
of cholesterol-fed rats rose to a peak in 2-4 weeks, then declined 
during the remainder of the experiment even though diets were kept 
constant. Serum phospholipid curves resembled cholesterol curves 
with initial rise and subsequent decline. In other studies (Hegsted, 
1957, 1959) it was found that in no instance did the addition of dietary 
cholesterol raise the phospholipid level above that of the control 
group, but it did raise serum cholesterol levels. By using various 
diets it was concluded that there is no great tendency for serum 
cholesterol to rise unless dietary cholesterol is added, and that this 
elevating effect is most pronounced if the cholesterol supplement is 
given in combination with a highly saturated fat-containing diet. 
Apparently homeostatic factors are involved, but they are least effec- 
tive in the presence of highly-saturated lipids. 


Relationship Between Chlosterol and Lecithin Intake 15 


In disagreement with these findings another group of investigators 
(Okey, et. al., 1951, 1959) found that after injecting cholesterol into 
rats for seven weeks the serum cholesterol values remained compar- 
atively constant, while the liver cholesterol esters were greatly in- 
creased, indicating that the dietary cholesterol was being held as an 
ester in the liver. They found that the concentration of free choles- 
terol in the livers was very small. Patil and Majar (1959) observed 
the slight increase in cholesterol content of serum and large increase 
in liver content for animals receiving cholesterol. 

It is believed by some that a large part of the circulating choles- 
terol is in combination in giant molecules, with protein, phospho- 
lipids, and fatty acids. If this is true then there is assumed to be a 
connection between the level of circulating cholesterol and phos- 
pholipids. 


Materials and Methods 


In the present study animals were fed two different lipid supple- 
ments, one of them being cholesterol which is the most common lipid 
found in the deposits in blood vessels. Having in mind that possibly 
blood serum cholesterol could be lowered with another lipid, lecithin 
was the other supplement. 

For this study forty-five young adult male rats (210gm-265gm) of 
a closely inbred albino strain were used. 

Normal serum lecithin and free cholesterol were determined for 
each animal and the determinations then averaged for final figures. 
These values were in close agreement with lipid values obtained 
by several oxidative micromethods. (Boyd, 1942) Blood was with- 
drawn via heart puncture from ether anesthetized animals. No anti- 
coagulant was used. The blood was allowed to clot and cenrtifuged 
at 2500 rpm for 20 minutes. A period of at least two weeks was 
allowed to elapse before blood was taken from the same animal again. 
All animals were kept at an even temperature and fed standard Purina 
lab chow plus Cod-liver oil, milk and cracked wheat bread. 

For the determination of free cholesterol one ml of serum was 
added to five ml of a 50-50 mixture of absolute ethyl alcohol and 
acetone, and the mixture then centrifuged. The filtrate was added to 
three ml of digitonin solution, which contained one drop of 10% 
acetic acid, and allowed to precipitate over night at room temper- 
ature in a closed preserving jar. The digitonin solution was prepared 
by dissolving 400 mg of digitonin in 100 ml of distilled water. Color 
was developed for the final determination by dissolving the digi- 
tonide in six ml of glacial acetic acid (Searcy and Berquist, 1950) 
saturated with ferrous sulfate, and then two ml of reagent grade 


16 Faye Johnson, Wm. G. Downs, Jr., and Robert L. Sublett 


concentrated sulfuric acid was added. The solution was diluted to 
ten ml with acetone-alcohol, and ten minutes were allowed for color 
development. Optical densities were measured on a Bausch and 
Lomb colorimeter at a wave length of 490 millimicrons, against a 
blank prepared with acetone-alcohol. 

Lecithin was determined by a modification of the Youngburg 
method in which inorganic phosphate was measuredfl One ml of 
serum was added to ten ml of mixture containing three parts ab- 
solute ethyl alcohol to one part anhydrous ether, and the resulting 
mixture brought to boiling in a water bath. The solution was filtered 
through filter paper which had been previously moistened with 
alcohol-ether. Five ml of the filtrate was evaporated to dryness and 
the residue then digested with 5 N sulfuric acid and 30% hydrogen 
peroxide. One ml of 2.5% ammonium molybdate solution, and 0.4 ml 
of aminonaphtholsulfonic acid was added to the solution, which was 
then diluted to ten ml with distilled water and allowed to stand five 
minutes for color development. Optical densities were measured on a 
Bausch and Lomb colorimeter at a wave length of 800 millimicrons, 
against a blank prepared with 10% trichloroacetic acid. 

The 2.5% ammonium molybdate solutoin was prepared by dis- 
solving 2.5 grams of ammonium molybdate in distilled water and di- 
luting to 100 ml. The aminonaphtholsulfonic acid reagent was pre- 
pared as follows: 0.5 grams of 1, 2, 4-aminonaphtholsulfonic acid was 
added to 195 ml of 15% sodium bisulfate solution. Five ml of 20% 
sodium sulfite solution was added and this was mixed thoroughly. 
Sodium sulfite was then added, one ml at a time, until the powder 
dissolved. This reagent was prepared fresh every two weeks. 

For experimental purposes the animals were equally divided into 
three groups. Group I received 125 mg cholesterol each, dissolved 
in corn oil; Group II received 125 mg lecithin each, dissolved in 
normal saline; and Group III received 125 mg cholesterol and 125 mg 
lecithin each, daily. All animals were fed the lipids orally. The 
animals were force fed with a pipette at first, but after a short time 
they readily ate the lipid supplements. The dosage was calculated 
on the basis of 0.5 gram per kilogram of body weight. 

Free cholesterol and lecithin determinations were made on blood 
from animals in each group at 3, 10, 19 and 26 days after the begin- 
ning of the experiment, according to the methods previously described. 


Results 


In contrast to the results obtained by some previous investigators 
these animals showed a response to increased lipid intake. Regardless 


Relationship Between Chlosterol and Lecithin Intake 17 


of the type of lipid being given, with every increase in lecithin con- 
tent, there was a decrease in free cholesterol content. This was also 
observed in the group receiving cholesterol and lecithin except for 
the determination made at 19 days in which the cholesterol content 
had dropped below normal, but rose on the 26th day. This latter 
rise cannot now be explained. (Chart I) 


giNormalji25Mg Cholesterol! |125Mg Lecithin 125Mg Lecithin and 
lo 125Mg Cholesterol 


MMB cholesterol 
(ALecithin 
3 10 19 26 10 19 26 3 On Tan LAS 


CHART 1 
BLOOD SERUM CONTENT OF LIPIDS IN RATS 


Discussion 


This study, in agreement with some and disagreement with other 
previous studies, indicates that rats do show a difference in serum 
levels with increased lipid intake. Factors such as water and food 
intake, and individual difference, should be considered as possible 
explanation of the deviations. 

It is now rather well established that the intake of unsaturated 
dietary fats reduce serum cholesterol. (Lewis, et. al., 1961) This 
decrease appears to be due to accelerated catabolism of cholesterol 
to bile acids, and to increased excretion of neutral sterols. The 
mechanism of action of unsaturated fat is probably to produce 
qualitative changes in circulating lipoproteins, which facilitates their 
catabolism and the elimination of the contained cholesterol, either 
as bile acids and neutral sterol, or possibly cholesterol handling at 
the cellular level. 


18 Faye Johnson, Wm. G. Downs, Jr., and Robert L. Sublett 


From the results obtained in this study several questions are pro- 
posed. Apparently there exists some antagonistic action between 
lecithin and cholesterol. Is there a definite correlation between the 
amount of free cholesterol and lecithin, as some have believed? If 
so, could hypercholesterolemia be prevented by an increased lecithin 
intake, or would the organism increase its capacity to handle this lipid 
too, after which it might not have any effect? The possibility of a 
homeostatic mechanism, which others have suggested, involving 
lecithin should also be considered. The latter seems likely since in- 
take of lecithin was observed to decrease cholesterol content. Also 
with increased cholesterol intake, there was observed an increase in 
lecithin content. Could there be a feed-back mechanism in which 
the amount of cholesterol is regulated by the amount of lecithin in 
the organism? In cases of hypercholesterolemia could there have been 
a prior inefficiency in the synthesis of lecithin? It is also quite pos- 
sible that high concentrations of lecithin are causng a change in the 
lipoproteins, which speed up their catabolism, and cholesterol is sub- 
sequently eliminated. 

Another point to consider is the belief by some workers that the 
main role of lecithin in the body is to aid in the transport of lipids, 
since lecithin is soluble in the body fluids. On this basis lecithin may 
decrease serum cholesterol by “carrying” it into the tissues, rather 
than having any effect on its metabolism. 


Summary 


Forty-five young adult albino rats of closely-inbred strain were 
divided into three equal groups. One group received orally 125 mg 
of cholesterol daily, the second 125 mg of lecithin daily, and the third 
group 125 mg of both lecithin and cholesterol. At weekly intervals 
and prior to feedings, determinations of the level of serum lecithin 
and free cholesterol were made on all animals. Animals receiving 
lecithin and those receiving both lipids showed a consistent reduc- 
tion in cholesterol levels with the exception of the final rise already 
noted. Increased water intake also decreased cholesterol level. Tech- 
niques used and a discussion of the possible explanations are included. 


References 

Boyd, Eldon M., 1942. Species variation in normal plasma lipids extimated by 
oxidation micro-methods. J. Biol. Chem. 143:131-132. 

Collens, William S., 1957. Variations in capacity of rabbits to handle ingestes 
cholesterol. Angiology. 8:6:513-519. 

Gofman, John W., et. al., 1950. The role of lipids and lipoproteins in atheroscler- 
osis. Sci. 111:166-171. 

Hegsted, D. M., 1957. The effect of various fats upon experimental hyper- 
cholesterolemia in the rat. J. Nutrition. 63;3:377-391. 


Relationship Between Chlostcrol and Lecithin Intake 19 


Hegsted, Mark, 1959. Comparative studies on the effect of dietary fats on serum 
cholesterol levels. Proceedings of Heinz International Symposium of Health. 
Fed., Proc. 18:2:52-54. 

Kreglow, Alan Frank, 1958. Control of serum cholesterol by experimental diets. 
J. Amer. Ger. Soc. -:2:134-139. 

Lewis, B., Pilkington, T. R. E. and Hood, K. A. 1961. A mechanism for the 
action of unsaturated fat in reducing the serum cholesterol. Clin. Sci. 
20:2:249-154. 

Okey, Ruth and Lyman, Marian M., 1957. Dietary fat and cholesterol metabolism. 
I Comparative effects of coconut and cottonseed oils and three levels of 
intake. J. Nutrition. 61:523-533. 

SS a ea , and Harris, Anne G., 1959. Effects of unsaturation of 
dhgtersy fats on liver and serum pice! Metabolism. 8:3:241-255. 

Patil, V. S. and Majar, N. G., 1959. Relation of dietary fat and cholesterol to 
tissue and serum cholesterol and polyunsaturated fatty acids in rats. Indian 
J. Med. Res. 47:4:448-455. 

Searcy, Ronald and Berquist, Lois, 1950. A new color reaction for the quantita- 
tion of serum cholesterol. Clin. Chim. Acta. 5:192-199. 

Seskind, Coleman R., et. al., 1959. Serum lipid analyses in rats fed natural and 
hydrogenated cottonseed oil with cholesterol and cholate. Proc. Soc. Exper. 
Biol. and Med. 102:1:190-95. 

it eee , et. al., 1959. Serum lipid levels in rats fed vegetable oils with 
and without cholesterol. Proc. Soc. Exper. Biol. and Med. 100:1:631-634. 


ON THE ACTIVITY OF PLETHODON GLUTINOSUS AS 
INFLUENCED BY LIGHT 


ROGER W. BARBOUR 
Department of Zoology, University of Kentucky 


Anyone who has done extensive collecting of salamanders could 
scarcely have failed to note the existence of a well-marked periodicity 
of many species. It is my purpose to here record a brief study of this 
periodism in one species, and the effects of various light conditions 
on this pattern of daily activity. 

In April and May, 1961, I determinued the times of activity of a 
series of 10 large, robust, and apparently healthy slimy salamanders, 
Plethodon glutinosus glutinosus Green, while exposing them to varying 
light conditions. 

Essentially, the experiment consisted in releasing the animals in a 
23 by 3 inch circular stainless steel tray, the bottom of which had been 
covered by dirt, dead leaves, and pieces of tree bark to simulate a 
forest floor. At hourly intervals for a period of 23 consecutive days the 
tray and its contents were photographed, using a Praktina FX 35 mm. 
camera with a motor drive and a Braun SP2 electronic flash unit. In- 
cluded in the photographs were also a Numechron Tymeter GMT clock 
and a Minneapolis Honeywell thermometer with its bulb and capillary 
tube inserted in the leaf mold within the tray. Two 100 watt bulbs in 
goose-neck lamps supplied the only light other than that of the flash 
unit. The bulbs were placed at opposite sides of the tray, two feet 
above the rim, and pointing toward the center of the tray. The lights 
were controlled by an Intermatic Time-all clock; the camera was acti- 
vated hourly by means of an Eagle Signal Co. Cycl-Flex timer. 

Examination of the negatives disclosed the number of salamanders 
on the surface of the leaf mold, the temperature within the container, 
and the time the photograph was taken. Animals were considered 
“active” if as much as the anterior third of the body was visible; other- 
wise they were judged “inactive.” Incidentally, in nearly every case 
the animal was either completely exposed or completely hidden. 

The experiment was divided into three parts; the termination of 
one part was followed immediately by the onset of the next. 

Part 1, Normal day. Lights were turned on at 6 a.m. and off 
at 6:00 p.m. Ten animals were introduced into the tray but one died 
after two days, a second died after five days. No additional animals 
were lost during the experiment. Part 1 ran for seven days, a total of 
65 animal/days. (One animal for one day equals one animal/day.) 


On the Activity of Plethodon glatinosus 21 

Part 2, Continuous dark. All lights continuously off, the only light 

being that of the electronic flash unit fring hourly, each flash with a 

duration of approximately 1/1000 second. Part 2 ran for five days, a 
total of 40 animal/days. 


Part 3, Continuous light. Lights on continuously; neither of the 
bulbs burned out, so the light was essentially constant for four days, 
a total of 32 animal/days. 


Part 4, Reverse day. The lights were turned off at 6:00 a.m. and 
on at 6:00 p.m., thus shifting the normal day-night rhythm 12 hours 
out of phase. This was continued for seven days, a total of 56 animal/ 
days. 

All data from the same hour in each part as outlined above were 
averaged. For example, all photographs taken at 9:00 a.m. during the 
five days of continuous dark were considered together, and the average 
number of individuals visible at this time was calculated. This was 
expressed as a percentage of the total number it would have been pos- 
sible to see were all active, (in this case, 40 animals) and the results 
plotted on a graph. (Fig. 1) 

Although this experiment has barely scratched the surface, there 
are certain conclusions that I feel may be safely drawn. 


80 


NORMAL DAY me savin 
TANT DARK ----- 

60 CONS 

CONSTANT LIGHT --eee- 

REVERSE DAY 


40 


PERCENT VISIBLE 


1200 
1400 2 
1600 
1800 


Oo oC ©] © 
Bee) §..3) 2 8 
Sie ie on tO, ou OL Ke 
TIME OF DAY 


Fig. 1.— Activity of Plethodon glutinos:s under varying light conditions 


bo 
bo 


Roger W. Barbour 


1. Plethodon g. glutinosus exhibits a well-marked circadian rhythm, 
being more active in darkness than in light. The number of active ani- 
mals increased as the dark period progressed. Number of active 
animals fell off sharply with the onset of the lighted period. 


2. Conditions of constant dark tend to inhibit activity during the 
normally dark hours, and to increase activity during the normally light 
hours. However, the normal pattern of activity remains well-marked. 


3. Conditions of constant light exert a great deal of influence in 
suppressing the normal rhythm, but it was still apparent under the con- 
ditions herein imposed. 


4. Temperature fluctuations on the order of 10 degrees Fahrenheit 
or less have little or no effect on the normal rhythm of Plethodon g. 
glutinosus. 

5. Under conditions herein imposed, it is possible to shift the daily 
rhythm of Plethodon g. glutinosus some 12 hours out of phase. In other 
words, by manipulation of light alone, individuals can be induced to 
become active when they would normally be quiescent, and vice versa. 


AN ANNOTATED CHECKLIST OF FISHES FROM DIX RIVER AND 
TRIBUTARIES (EXCLUSIVE OF HERRINGTON RESERVOIR) 


PHILLIP E. GREESON 
Department of Zoology, University of Kentucky 


Introduction 


A study in the fish population dynamics of Herrington Reservoir 
by Whitney (1962) instigated a study of the fish fauna of the Dix 
River and its tributaries. Twenty-seven collecting stations were ran- 
domly selected throughout the system and collections were conducted 
during the summer months (June, July, August and September) of 
1962. 

The checklist includes the fishes taken with seines (4, 6 and 15 feet 
in length) and with gill nets (100’x6’ with 114” mesh). The classifica- 
tion by families, genera, and species is according to Moore in Blair 
et al. (1957). The common names are those suggested by the Ameri- 
can Fisheries Society (1960). 


Dix River and Tributaries 


The Dix River, a branch of the Kentucky River, is located in the 
southern portion of the Bluegrass region of Central Kentucky. It ex- 
tends a distance of 78 miles through Mercer, Boyle, Garrard, Lincoln, 
and Rockcastle counties. An area of 449 square miles composes the 
watershed. 

Even though the Dix River system is relatively small, it offers three 
distinct ecological habitats; a shallow river system, a deep impound- 
ment, and a cold tailrace. The shallow river system, the concern of this 
article, consists of two primary, complexly-branched systems: the Dix 
River proper and the Hanging Fork Creek system. These two systems 
join at the extreme head of Herrington Reservoir. 

The water depth ranges from a few inches in the head waters to 
four feet at the reservoir junction. The relatively steep gradient (about 
5.9 feet per mile) combined with this water shallowness produces 
riffles and areas of swift current. The flow is interrupted during pe- 
riods of drought and periodical pools result ranging in dept from two 
inches to two feet. This is typical of the summer months. Flash floods 
are common during the spring and fall rainy seasons. 

Overburden of the basin is composed of thin, rocky materials on 
the hilltops; the slopes of the more rugged sections are of rich lime- 
stone soils. In the source region (the edge of the Knobs), tight and 
clayey materials are found in the lowlands (Whitney, 1962). 


24 Phillip E. Greeson 


Results of Collections 
Clupeidae 
Dorosoma cepedianum (LeSueur). Twenty-one specimens of the gizzard 
shad, an important forage fish, were collected from the main trunk stream. Two 
individuals were taken from Hanging Fork Creek. 


Esocidae 


Fsox vermiculatus Le Sueur. No specimens of the grass pickerel were col- 
lected but a single individual, taken from the extreme headend of the Dix River 
proper, exists in the collections of the Department of Zoology, University of Ken- 
tucky. It was taken in October, 1960. 


Catostomidae 


Carpiodes cyprinus (LeSueur). Three young quillbacks were collected at the 
junction of the river and the impoundment. 


Catostomus commersoni (Lacépéde). One specimen was obtained from a 
shallow headwater stream, but the greatest abundance of the white sucker ap- 
peared to be in the deeper portions of the main trunk and in the main stream of 
Hanging Fork Creek. 

Hypentelium nigricans (LeSueur). Five hog suckers were collected from the 
deeper pools of the shallow streams and eleven individuals were taken at various 
points along the main trunk. 


Moxostoma breviceps (Cope). The shorthead redhorse was the typical 
sucker of the system. Twelve specimens were taken from headwater streams. Their 
relative abundance increased as the streams became larger. 


Moxostoma erythrurum (Rafinesque). The golden redhorse was obtained 
from intermediate-size streams but its greatest abundance was found in the Dix 
River proper. Thirty-six individuals were captured. 


Cyprinidae 

Campostoma anomalum (Rafinesque). A total of six hundred and thirty-seven 
stonerollers were collected throughout the entire system. Their relative abundance 
was greater in headwater and intermediate-size streams. 


Chrosomus erythrogaster Rafinesque. The southern redbelly dace was collected 
from only one headwater stream. Two specimens were obtained. 


Ericymba buccata Cope. One hundred and fifty-four individuals were col- 
lected from intermediate-size streams. An occasional silverjaw minnow was col- 
lected from the main trunk stream. 


Hybopsis amblops (Rafinesque). The deeper water of the Dix River proper 
yielded seven specimens of the bigeye chub. 


Notropis ardens (Cope). Two hundred and seventeen rosefin shiners were 
taken from the deeper pools of the intermediate-size streams and from the trunk 
stream of Hanging Fork Creek. 


Notropis atherinoides Rafinesque. A moderate abundance of the emerald 
shiner characterized the pools of the main trunk stream and the trunk of the 
Hanging Fork Creek. 


Notropis boops Gilbert. The bigeye shiner was collected from all portions of 
the main trunk stream and the lower portions of the larger tributaries. One hun- 
dred and twenty-five specimens were obtained. 


Notropis cornutus (Mitchill). The common shiner, like the bigeye shiner, 
was found in all portions of the main trunk stream and the lower portions of the 
larger tributaries. One hundred and thirty-one specimens were collected. 


Notropis rubellus (Agassiz). Thirteen rosyface shiners were obtained from a 
deeper section of Hanging Fork Creek. 


An Annotated Checklist of Fishes 25 


Notropis spilopterus (Cope). Twenty spotfin shiners were collected from the 
lower end of both the main truck stream and Hanging Fork Creek. 

Notropis whipplei (Girard). A moderate abundance of the steel-color shiner 
was obtained from the deeper waters of the river. Fifty-seven individuals were 
collected. 

Pimephales notatus (Rafinesque). The bluntnose minnow was collected 
throughout the entire river system, but its greatest abundance was in the inter- 
mediate-size streams. Very few specimens were taken from Hanging Fork branch. 

Pimephales promelas Rafinesque. Only three fathead minnows were collected. 
They appeared in a pool of a single headwater branch. 

Semotilus atromaculatus (Mitchill). The creek chub, the characteristic 
minnow-form of the river, was collected from all stations. Hundreds were captured 
but only five hundred and fifteen were retained. 


Ictaluridae 


Ictalurus melas (Rafinesque). A single specimen of the black bullhead, ex- 
pected to be restricted to the deeper waters, was netted in a deep pool of a larger 
headwater stream. Several spcimens were collected from the main trunk stream. 

Ictalurus natalis (LeSueur). Three yellow bullheads were obtained from the 
Dix River proper. % 

Ictalurus punctatus (Rafinesque). A single channel catfish was recorded at 
the junction of the river and the reservoir. 

Noturus miurus Jordan. A single brindled madtom was collected about mid- 
way in the main trunk stream. 


Atherinidae 


Labidesthes sicculus (Cope). Collections from Hanging Fork Creek and the 
main trunk stream revealed nine brook silversides. 


Serranidae 


Roccus chrysops (Rafinesque). Only two young specimens were netted in 
the river. According to Tompkins and Peters (1951), the river becomes the site 
for the annual spawn runs of the white bass. 


Centrarchidae 


Ambloplites rupestris (Rafinesque). A total of five rock bass were collected 
from four stations located on the main trunk stream and the Hanging Fork Creek. 


Lepomis cyanellus Rafinesque. The green sunfish extended into the extreme 
headwaters and its greatest abundance was in the intermediate-size streams. A 
total of twenty-six specimens were netted. 


Lepomis macrochirus Rafinesque. Thirty-three bluegill individuals were ob- 
tained from the system. They were found in the lower, pooled portions of the 
headwaters and extended to the lake junction. 


Lepomis megalotis (Rafinesque). The longear sunfish was collected in the 
intermediate-size streams buts its greatest abundance appeared to be in the deeper 
waters. Twenty-one specimens were netted. 

Micropterus dolomieui (Lacépéde). Two young specimens of the smallmouth 
bass were collected in a deep pool in Hanging Fork Creek. 

Micropterus punctulatus (Rafinesque). Single specimens were collected from 
each of four stations. The spotted -bass was taken only in the deeper waters. 

Micropterus salmoides (acépéde). Five largemouth bass were netted in 
the deeper waters. On one occasion, a single specimen was captured in an inter- 
mediate stream. 


26 Phillip E. Greeson 


Pomoxis annularis Rafinesque. Eight specimens of the white crappie were 
obtained in the main trunk stream and one specimen from Hanging Fork Creek. 


Pomoxis nigromaculatus (LeSueur). The black crappie is rare in the Dix 
River. Only one specimen was collected in Hanging Fork Creek. 


Percidae 


Etheostoma blennioides Rafinesque. Thirty-three greenside darters were ob- 
tained from swift riffle areas of the Dix River. Two specimens were collected in 
riffles of the lower Hanging Fork Creek. 


Etheostoma caeruleum Storer. The rainbow darter appeared to have an even 
distribution throughout the main stream and the lower ends of tributaries. Twenty- 
four specimens were recorded. 


Estheostoma flabellare Rafinesque. The fantail darter appeared to be the 
most stream diversified species. It was collected from every station. Thousands 
were captured but only one hundred and ninety-five were retained. 


Ethecstoma nigrum Rafinesque. The johnny darter was collected from riffles 
in the main trunk and in the intermediate-size streams. Its greatest abundance was 
in the latter. 


Etheostoma spectabile (Agassiz). Six orangethroat darters were captured in 
lower headwater riffles and two in a main trunk riffle. This strange separation 
might indicate overlooked individuals in between. 


Percina caprodes (Rafinesque). The greatest abundance of the log perch 
was in the deeper portions of the main trunk, but it extended to the lower margins 
of intermediate-size streams. A total of seventeen specimens were collected. 


Percina macrocephala (Cope). The longhead darter is very rare in the sys- 
tem. A single specimen was obtained at the junction of the river and the lake. 


Sciaenidae 


Aplodinotus grunniens Rafineque. Two freshwater drums were collected from 
the deeper portion of the main trunk. 


Discussion 


The fishes collected from twenty-seven stations in the Dix River 
system represented forty-four species from ten families. The specific 
distributions were quite uniform and distinct patterns were revealed. 

The suckers were the predominant rough fish and were collected 
throughout the enitre system. Their greatest abundance was in the 
deeper portions of the Dix River and Hanging Fork Creek. 

The intermediate-size streams were characterized by the abundance 
of minnows. The genus, Notropis, appeared to be restricted to these 
waters where seven species were netted. The creek chub, the blunt- 
nose minnow, and the stoneroller were the only family representatives 
extending into the headwaters; but, even here, they were not charac- 
teristic. 

The deeper waters at the lower end of the river proved to be a 
favored habitat for all the collected species of catfish. The Centrar- 
chidae (as well as the freshwater drum, the brook silversides, and the 
crappie) were also primary inhabitants of this area. 


An Annotated Checklist of Fishes 27 


The most pronounced distribution pattern was exhibited by the 
sunfishes of the genus Lepomis. The green sunfish was collected 
throughout the entire river system. The bluegill was collected from 
all streams except those of the extreme headwaters. The longear sun- 
fish made its first appearance in the intermediate-size streams and its 
relative abundance increased as the water depth increased. 

Darters were collected from practically all riffled areas but mem- 
bers of the genus Etheostoma had their greatest numbers in the head- 
waters and the intermediate-size streams. The log perch was distinc- 
tive of the deeper waters. 


Summary 


1. Following a fish population study of Herrington Reservoir 
(Whitney, 1962), a study was undertaken to determine the fish fauna 
of the Dix River and tributaries. 


2. Twenty-seven collecting stations were established and collec- 
tions were made during a four month period in 1962. 


3. The findings are discussed with regards to the family representa- 
tives and relative distributions. 


Literature Cited 


American Fisheries Society. 1960. Special Publication, No. 2. A list of common 
and scientific names of fishes from the United States and Canada. Ann 
Arbor, Mich. 102 pp. 

Blair, W. F., et al. 1957. Vertebrates of the United States. Mc-Graw-Hill Book 
Co. New York. 819 pp. 

Clay, W. M. 1962. A Field Manual of Kentucky Fishes. Ky. Dept. of Fish and 
Wildlife Resources. Frankfort. 147 pp. 

Tompkins, W. A. and M. M. Peters. 1951. The age and growth of the white bass, 
Lepibema chrysops, of Herrington Lake, Kenutcky. Ky. Fish Bull. No. 8. Ky. 
Div. of Fisheries. 15 pp. 

Turner, W. R. 1953. The age and growth of the gizzard shad, Dorosoma cepe- 
dianum (LeSueur), in Herrington Lake, Kentucky. Ky. Fish. Bull. No. 13. 
Ky. Div. of Fisheries. 15 pp. 

Whitney, E. W. 1962. Investigations and management of the fisheries of Herring- 
ton Lake and Dewey Lake, Kentucky. Ky. Dept. of Fish and Wildlife Re- 
sources. Frankfort. 74 pp. 


AGING BATS IN WINTER 


WAYNE H. DAVIS 


Department of Zoology 
University of Kentucky, Lexington 


A method for separating young bats from old ones in winter has 
long been desired by bat banders studying survival and longevity. 
Recent observations on Pipistrellus subflavus have suggested that some 
young of the year can be recognized in hibernating populations in 
certain areas by examining epiphyses and pelt. 

In those populations of Pipistrellus subflavus which winter in Ver- 
mont, Quebec and eastern Ontario young of the year can easily be 
separated from the older bats. The epiphyses of the young have not 
closed by the time they enter the caves for hibernation. Apparently 
no growth of the epiphyses takes place after the bats enter the caves, 
for young are as readily distinguished in April and May as in October. 
This is based upon examination of 33 study skins from the above areas. 
Farther south, in the caves of West Virginia and Kentucky, most young 
have closed epiphyses by the time they enter hibernation. A few 
individuals, however, are recognizable (Plate I B). This is also true 
of those wintering in southern Wisconsin and Minnesota. 

Juvenile pipistrelles in West Virginia molt in the autumn. In the 
northeastern part of their range young pipistrelles go into hibernation 
before molting. Some of those taken from the caves of Vermont in the 
early winter have a juvenile pelage which is no different from that of 
young shot in West Virginia in August and September (e.g. Natl. 
Mus. nos. 296717 & 296718, from East Dorset, Bennington County, 
Vermont, Dec. 12, 1948). The juvenile pelage is recognizable in that 
it is darker, particularly the subterminal band of the hair, and the 
banding is not as distinctive as on the adults. 

Pipistrelles wintering in Vermont are probably born in early July, 
and may be little over two months old when entering the caves for 
hibernation. At the other extreme are those pipistrelles which hiber- 
nate in the caves at Florida Caverns State Park in Jackson County, 
Florida. Among 14 skins examined from this population, young of the 
year could not be distinguished. The epiphyses are closed and the 
first molt has been completed in all individuals. These bats probably 
do not go into hibernation until late in the autumn. 

Since the molt has not begun when the young pipistrelles of Ver- 
mont go into hibernation, and is completed before hibernation by 


Aging Bats in Winter 29 


those in Florida, there should be an intermediate area in which some 
young have completed molting and some have not when taken from 
the caves in autumn. A series of 43 pipistrelles taken from the caves 
in Carter Caves State Park, Carter County, Kentucky, during the 
autumn and winter months was examined for signs of molt. Three 
individuals were found which apparently were in the process of molt- 
ing when they entered the caves for hibernation. All of these were 
young of the year. One is of particular interest in that most of the 
juvenile pelage is still apparent, and the new fur can be seen coming 
in at the base of the old in the dorsoposterior region (Plate I D). The 
pelt in this region is unprime. That the bat was quite young when 
entering hibernation is evident from the epiphyses (Plate I B). 


Photos by Luis de la Torre 


PLATE | Pipistrellus subflayus 
A.—Individual very young at the time of entering hibernation (Amer. Mus. No. 100161, 
from a cave at Rutland, Chittenden Co., Vt., Nov. 8, 1931). 
B.—Young bat, somewhat older than A at the time of entering hibernation (W. G. Frum 
No. 1479, from Carter Cave, Carter Co., Ky., Nov. 12, 1950). 
C.—Adult (W. G. Frum No. 2308, from Cave Mt. Cave, Pendleton Co., W. Va., Dec. 31, 
1954). 


D.—Same bat as B, showing juvenile molt in posterior region. Juvenile pelage on head 
and shoulders, 


30 Wayne H. Davis 


Acknowledgments.—For the loan of specimens used in this study 
the writer wishes to thank F. L. Burnett, Museum of Comparative 
Zcology, Harvard University; W. G. Frum; G. G. Goodwin, American 
Museum of Natural History; F. L. Osgood; and V. S. Schantz, Biologi- 
cal Surveys Collections and U.S. National Museum. 


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STEELE, DEWEY G., Animal Industry Dept., University of Kentucky, Lexington, Ky. Biol. 
STEWART, ORVILLE W., 125 Johnston Blvd., Lexington, Gy. 

STRICKLER, THOMAS D., Physics Dept., Berea College, Berea, Ky. 


SUMPTER. WARD C., Chemistry Dept., Western Kentucky State College, Bowling Green, 
Ky. Chem. 


TABB, MARION F., R.F.D. No. 4, Hillsboro, Ohio. 

TAYLOR, RALPH W., 4146 Monroe St., Louisville 13, Ky. 

TESSENEER, RALPH A., Psychology Dept., Murray State College, Murray, Ky. 
THOMPSON, HERBERT H., JR., 609 So. University Dr., Fairfax, Virginia. 
THORPE, JAMES F., 1145 Claridge Dr., Lexington, Ky. Eng. 

TODD, JACK R., Agronomy Dept., University of Lentucky, Lexington, Ky. 
TODD, ROBERT E., JR., Route 1, Box 316, Jeffersontown, Ky. 


TOWNSEND, LEE HILL, Entomology and Botany Dept., University of Kentucky, Lexington, 
Ky. Biol. 


TRUE, JOHN W., 1925 So. Third St., Louisville 8, Ky. Biol. 
TYNER, H. D., 323 Prentiss Ave., Findlay, Ohio. 


WADE, CLAUDE F., Dept. Conservation, Entomology Division, State Office Bldg, Indianapolis, 
Ind. 


WAGNER, WILLIAM F., Chemistry Dept., University of Kentucky, Lexington, Ky. Chem. 


WALTON, WARREN W., JR., Dept. Mechanical Engineering, University of Kentucky, Lexing- 
ton, Ky. 


WEAVER, R. H., Bacteriology Dept., University of Kentucky, Lexington, Ky. Bact. 
WEBER, EDWARD H., Bellarmine College, Louisville, Ky. 

WELDON, J. ELMER, Box 531, 127 Jackson St., Georgetown, Ky. Psych. 

WELLS, JOHN B., JR., Mathematics Dept., University of Kentucky, Lexington, Ky. 
WHARTON, MARY E., Biology Dept., Georgetown College, Georgetown, Ky. 
WHEDBEE, ELLIS D., JR., 2832 W. Chestnut St., Louisville 11, Ky. 

WHITE, M. M., University of Kentucky, Lexington, Ky. Psych. 

WHITNAH, ROBERTA, Box 966 College Station, Murray, Ky. 

WHITT, ALLIE L., JR., Biology Dept., Eastern Kentucky State College, Richmond, Ky. 
WHITTLE, CHARLES E., Physics Dept., Western Kentucky State College, Bowling Green, Ky. 
WIKLER, ABRAHAM, 1241 Summit Dr., Lexington, Ky. 

WILEY, R. H., 210 Pleasantview Ave., Louisville 6, Ky. Chem. 


WILLIAMS, GORDON C., Dept. Chemical Engineering, University of Louisville, Louisville, 
Ky. Chem., Eng. 


WILLIAMS, MRS. HARRIET, 269 Cassidy, Lexington, Ky. 

WILLIAMS, W. L., P. O. Box 5125, Louisville 5, Kentucky. Bact. 

WILSON, GORDON, 1434 Chestnut St., Bowling Green, Ky. Biol. 

WISEMAN, RALPH F., Bacteriology Dept., University of Kentucky, Lexington Ky. Bact. 
WITTIG, C.O.G., University of Louisville, Louisville, Ky. Eng. 

WOLFE, OTIS K., JR., Physics Dept., Centre College, Danville, Ky. 

ZIMMACK, HAROLD L., Biology Dept., Eastern Kentucky State College, Richmond, Ky. Biol. 


SUSTAINING: MEMBERS 


ASHLAND OIL AND REFINING COMPANY, Ashland, Ky. (5) 

BRAUER, ALFRED, Zoology Dept., University of Kentucky, Lexington, Ky. Biol. 
CANTRELL, WILLIAM F., Pharmacology Dept., University of Louisville, Louisville, Ky. 
CORHART REFRACTORIES CO., 940 Commonwealth Bldg., Louisville 2, Ky. 

FOHS, F. J., 1077 San Jacinto Bldg., Houston, Texas. 

KENTUCKY STATE COLLEGE, Frankfort, Ky. 

KENTUCKY UTILITIES COMPANY, Lexington, Ky. (11). 

MILES, PHIL M., 229 Barrow Rd., Lexington, Ky. 

MILETI, OTTO J., Mileti Company, Inc., 1401 So. 15th St., Louisville, Ky. 

MURRAY STATE COLLEGE, Murray, Ky. 

PENROD, E. B., Dept. of Mechanical Engineering, University of Kentucky, Lexington, Ky. 
SPANYER, J. WILLIAM, Brown-Forman Distillers Corp., Louisville, Ky. 

VALLEAU, W. D., Agronomy Dept., University of Kentucky, Lexington, Ky. 

WESTERN KENTUCKY STATE COLLEGE, Bowling Green, Ky. 

WOLFSON, ALFRED M., 310 N. 14th St., Murray, Ky. 


36 Membership List 


LIFE MEMBERS 


GRAHAM, THOMAS, Bankers’ Bond Co., Kentucky Home Life Bldg., Louisville 2, Ky. 
HEUSER, HENRY V., Henry Vogt Machine Co., Louisville, Ky. 

JILLSON, W. R., 301 W. Third St., Frankfort, Ky. 

PENNEBAKER, G. B., Tennessee Polytechnic Institute, Cookeville, Tenn. 

PURYEAR, ROBERT, Bankers’ Bond Co., Kentucky Home Life Bldg., Louisville, Ky. 
SPANN, LIZA, Murray State College, Murray, Ky. 

WILSON, WILLIAM S., Box 18, College, Alaska. 


INDUSTRIAL MEMBERS 
PHILLIP MORRIS INC., Louisville, Ky. (2) 


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