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Rt Brera cn bea agen rte a Saath wake ts tae tbe ait Cheeta g vam iit ars year i sata eas Loy ne ea Pens ing ae es Lines ght i ae ur Magy oF MPa earnest i tee OF BY Pen 4 ay att uy ia bie aT 1H ev mat oaks Ps re Chak os ae re feat 38 uA iad is ote bathe ty iat + ea $n ies aerate Cee Lik tj c HY Vee SAIN oe sh bd ght ron ve err ae COs “at Si ete tn wate Aga etry uag ye cepety awsyt acest tae Peake es a eae, BF Braet Wo died uy ane Patt Vea 24 er cried egieay peel gna aa eye i Patent a ty, dae be yale Fatt ty tue seen Wb I fi bs ohne he a ie be eben tye rtd telus poet " ‘ eh ort satotatcles yee Mi cs sits oy elem Pmeyee iy ae anes *TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE . i , hi ‘, : Volume 56 Numbers 1-2 . March 1995 ( Official Publication of the Academy The Kentucky Academy of Science Founded 8 May 1914 Governinc Boarp For 1995 ExecuTivE COMMITTEE President: Robert Creek, Department of Biological, Eastern Kentucky University, Richmond, KY 40475 President Elect: William S. Bryant, Department of Biology, Thomas More College, Crestview Hills, KY 41017 Vice President: Marcus T. McEllistrem, Department of Physics, University of Kentucky, Lexington, KY 40506-0055 Past President: Larry P. Elliott, Department of Biology, Western Kentucky University, Bowling Green, KY 42101-3575 Secretary: Peter X. Armendarez, Department of Chemistry and Physics, Brescia College, Owensboro, KY 42301 Treasurer: Julia H. Carter, Wood Hudson Cancer Research Laboratory, 931 Isabella Street, Newport, KY 41071 Executive Secretary (ex officio): J. G. Rodriguez, Department of Entomology, University of Kentucky, Lexington, KY 40546-0091 Editor, TRANSACTIONS (ex officio): Branley A. Branson, Department of Biological Sciences, Eastern Kentucky University, Richmond, KY 40475 Editor, NEWSLETTER (ex officio): Vincent DiNoto, Natural Science Division, Jefferson Community Col- lege, Louisville, KY 40272 MEMBERS, GOVERNING BOARD Blaine R. Ferrell 1995 Wimberly C. Royster 1997 Valena Hurt 1995 James Hopgood 1998 Patricia K. Doolin 1996 Robert H. Wombies 1998 David E. Hogan 1996 AAAS/NAAS Representative Gerald L. DeMoss 1997 J. G. Rodriguez 222? Chairperson, KJAS Valgene L. Dunham 1995 COMMITTEE ON PUBLICATIONS Editor and Chairman: Associate Editor: Abstract Editor: Index Editor: Editorial Board: Branley A. Branson, Department of Biological Sciences, Eastern Kentucky University, Richmond 40475 Thomas K. Green, Department of Chemistry, Western Kentucky University, Bowling Green 42101 Robert F. C. Naezi, Department of Biological Sciences, Northern Kentucky University, Highland Heights 41076 Varley Weideman, Department of Biology, University of Louisville, Louisville 40292 Vincent DiNoto, Natural Science Division, Jefferson Community College, Louisville 40201 Jerry N. Baskins, School of Biological Sciences, University of Kentucky, Lexington 40506 John P. Harley, Department of Biological Sciences, Eastern Kentucky University, Richmond 40475 Robert Creek, Department of Biological Sciences, Eastern Kentucky University, Richmond 40475 Larry P. Elliott, Department of Biology, Western Kentucky University, Bowling Green 42101 John Thioret, Department of Biology, Northern Kentucky University, Highland Heights 41076 All manuscripts and correspondence concerning manuscripts should be addressed to the Editor. Authors must be members of the Academy. The TRANSACTIONS are indexed in the Science Citation Index. Coden TKASAT. ISSN No. 0023-0081. Membership in the Academy is open to interested persons upon nomination, payment 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. Annual dues are $25.00 for Active Members; $15.00 for Student Members; $35.00 for Family; $350.00 for Life Mem- bers. Subscription rates for nonmembers are: domestic, $45.00; foreign $50.00; back issues are $30.00 per volume. The TRANSACTIONS are issued semiannually in March and September. Four numbers comprise a volume. Correspondence concerning memberships or subscriptions should be addressed to the Secretary. Exchanges and corre- spondence relating to exchanges should be addressed to the Librarian, University of Louisville, Louisville, Kentucky 40292, the exchange agent for the Academy. This paper meets the requirements of ANSI/NISO Z39.48-1992 (Permanence of Paper). 6 ee ee ee 7 : : ° y uf +o : ie (Al tue, 4, a 1 on ate Paes » pe be Sis. sear ; ' yin bi 4 ; rei ot ie ’ “a *Pts4 oe: vere MS a ‘ oe a na —— 1 eae ° 9 7 ae wapue Ap & ‘ win b>. ee oe a ay 1°, 8 si brew A = =. i pres at 1 GAG y * Ws “4 % 5H 7 — ’ on " F yt" 9 €_ re ee i) er 7 Lane Cells is . 7 a) ee ‘ f ta a J e. lca >, Chili 1 serena.) cs + web e Ge ii i Duh: peri € Ldoniicauity 7 Py) iP) eee wide Vor «| cathe, = tess, ~~ ithe - iu a of a INDUSTRIAL AFFILIATES ASSOCIATE PATRON ASHLAND OIL, INC. MEMBER CoORHART REFRACTORIES CORP. MPD, Inc. UNITED CarTAtyst, INC. ASSOCIATE MEMBER 3M TarE MANUFACTURING DIVISION, CYNTHIANA PLANT ALL-RITE PEsT CONTROL HoerEcuHst CELANESE CorpP.-ENGINEERING PLASTICS DIVISON KENTUCKY SCIENCE & TECHNOLOGY COUNCIL, INC. WESTVACO INSTITUTIONAL AFFILIATES FELLOW University of Kentucky SUSTAINING MEMBER Eastern Kentucky University Morehead State University Murray State University Northern Kentucky University University of Louisville Western Kentucky University MEMBER Campbellsville College Cumberland College Kentucky State University Paducah Community College Southeast Community College ASSOCIATE MEMBER Ashland Community College Berea College Brescia College Centre College Georgetown College Hazard Community College Jefferson Community College Kentucky Wesleyan College Lees College Maysville Community College Midway College Owensboro Community College Somerset Community College Spalding University Thomas More College Transylvania University TRANSACTIONS of the KENTUCKY ACADEMY of SCIENCE March 1995 Volume 56 Numbers 1-2 Trans. Ky. Acad. Sci., 56(1—-2), 1995, 1-8 Changes in Vegetation and Bobwhite Quail and Eastern Cottontail Rabbit Use in a Converted Fescue Field JEFFERY D. SOLE Upland Game Program, Kentucky Department of Fish and Wildlife Resources, I g : I Frankfort, Kentucky 40601 ABSTRACT A pair of approximately 8.1 ha (20 acre) tall fescue (Festuca arundinacea) dominated fields at Kleber Wildlife Management Area (KWMA), Owen County, KY, were chosen to determine the effects of converting fescue-dominated vegetation to an orchard grass/legume mixture on bobwhite quail (Colinus virginianus) and eastern cottontail rabbit (Sylvilagus floridanus) habitat and utilization. One field was treated and 1 remained untreated for comparison. The treated field showed increased plant species diversity (66 spp. on untreated/101 spp. on treated), a higher percentage bare ground the year of treatment (10.4% on untreated/ 24.4% on treated), and a greater abundance of legumes (6 spp. of legumes on untreated, none having > 1.0% total cover/8 spp. of legumes on treated, 6 having >1.0% total vegetative cover). The treated field had 27 plant species providing at least 1.0 per cent cover while the untreated field had 10 species. An index value was calculated to compare the value of the vegetation of the fields to bobwhite quail and eastern cottontails. The treated field was rated at 59 points and the untreated field rated at 14 points (the higher the index value, the better the habitat). Quail and rabbit use of the treated field increased while the untreated field remained unutilized. Costs involved to implement the fescue conversion and mosaic mowing were $182.15/ ha plus approximately 20 manhours/ha labor. INTRODUCTION Eastern cottontail rabbit and northern bob- white quail are 2 of the most important game species in Kentucky. Recent data indicate the eastern cottontail rabbit to be the third most sought-after game species in the state with ap- proximately 160,000 rabbit hunters harvesting 1.5 million cottontails annually (1, 2). Rabbit hunting provides an estimated 1.4 million mandays of recreation to Kentucky sportsmen while contributing about 23.7 million dollars to local economies (1, 2). The northern bob- white quail is the number | game bird in Ken- tucky and ranks 6th in popularity as a Ken- tucky game species (1). Northern bobwhite quail and eastern cot- tontail rabbit populations have shown long- term declines in Kentucky over the last 3 de- cades (3). These population trends are gen- erally attributed to habitat losses due to major changes in land-use practices such as urban sprawl, intensification of commercial agricul- tural tillage practices, and widespread conver- sion of open lands, pastureland and hayland to “KY 31” tall fescue (Festuca arundinacea) dominated stands. Of the approximately 7.0 million acres of grassland in Kentucky, 5.5 mil- lion acres have been planted to “KY 31” tall fescue cover (4). Wildlife biologists have long recognized the low wildlife habitat value provided by dense sod forming grasses such as “KY 31” tall fes- cue (5-13). The highly competitive and inva- sive growth of “KY 31” results in reduced plant : species diversity within hayland/pastures and old fields. The dominance of “KY 31” over other pasture and old field species has result- ed in an almost uniform coverage of Kentuc- 2 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-2) ky’s open lands. A second problem noted with “KY 31” is the thick, rank growth habit char- acteristic of the species which restricts move- ments of wildlife species and retards plant suc- cession. More recent evidence has revealed a third problem associated with KY 31, or more ap- propriately, with the endophytic fungus (Acre- monium coenophialum) which lives in “KY 31”. Lacefield et al. (14) and Siegel et al. (4) reported nearly 80% of the tall fescue fields in Kentucky are infected with the endophyte, at a rate of 80% or higher. Data indicate a diet of endophyte-infected “KY 31” impacts the re- productive potential of laboratory animals and livestock (i.e., 15-21) through decreased sperm and egg counts, smaller than normal lit- ter sizes, lowered lactation rates, poor weight gains, elevated body temperatures, and abor- tion or absorption of fetuses. Sadler (8) found cottontail rabbit reproduction and_ survival were reduced when rabbits were kept in out- door pens vegetated with tall fescue. Betsill et al. (7) reported cottontails in North Carolina avoided areas containing fescue. Recognizing that lowered reproductive po- tential reported in laboratory animals and live- stock may apply to wild cottontails and other wildlife species living in fescue-dominated habitat, the Kentucky Department of Fish and Wildlife Resources (KDFWR) recommends conversion of “KY 31” tall fescue fields to oth- er cover types by private landowners interest- ed in improving wildlife habitat on their land. Terrain often dictates no-till conversion meth- ods be used. In conjunction with fescue con- version, KDFWR often recommends mowing practices be implemented to improve inter- spersion of escape cover. This study was initiated to determine the effect of converting “KY 31” tall fescue-dom- inated fields to another grass/legume mixture, coupled with mosaic pattern mowing to im- prove escape-cover interspersion, on northern bobwhite quail and eastern cottontail rabbit populations. The costs associated with no-till conversion of tall fescue were also document- ed. STUDY AREA The Kleber Wildlife Management Area is located approximately 35.4 km northeast of Frankfort, within the Outer Bluegrass phys- iographic region, in Owen and Franklin coun- ties, Kentucky. This 929 ha area is character- ized by steep rolling hills with narrow flat ridge tops and narrow stream valleys. Eleva- tions range from 198 to 274 m msl. Approxi- mately 60% of the area is wooded, with east- ern red cedar (Juniperus virginianus) domi- nated old fields and woodlands and oak-hick- ory dominated woodlots. The remaining 40% of the area is old-field type open lands domi- nated by tall fescue. Kleber Wildlife Manage- ment Area (KWMA) was chosen as a study site because habitat conditions were very similar to those typically found on privately owned land in many parts of the state. MATERIALS AND METHODS Two fescue-dominated fields (1 control, 1 conversion), each about 8.1 ha in size, were chosen for study on KWMA. Baseline data documenting bobwhite quail and cottontail rabbit use and vegetative composition were gathered for both fields for 1-year prior to treatment (1987). A fescue conversion project was implemented on 1 field in 1988. Vegeta- tive composition and quail and rabbit use were monitored on both fields through 1990. Vegetative Sampling.—Vegetative sampling was conducted annually on both fields during September. These data were collected 1 year prior to treatment (1987) and for 3 years fol- lowing treatment (1988-1990). Forty 1-m? sample plots were positioned at 30.5 m inter- vals along line transects. Transects were de- signed to bisect all slopes and aspects present and were marked with steel fence posts. A compass bearing from | post to the other was followed to make the yearly sample points as consistent as possible. Ocular estimates were made of per cent total vegetative cover, per cent bare ground, average vegetation height, and per cent total cover contributed by each plant species. Individual species were ranked by per cent total cover contributed in each treatment type. Plant species with >4.0% total cover were as- signed a subjective numerical rating of good (=2), fair (=1) or poor (=0) as food and cover for quail and rabbits based on literature re- view (ie., 5, L1-13, 22-41). An overall nu- merical index value for each field was derived by summing index values for each plant spe- cies providing at least 4.0% total cover. CHANGES IN VEGETATION, QUAILS, Quail and Rabbit Sampling.—Flush-drive censuses (42) were conducted in September for | season prior to treatment (1987), during the treatment year (1988) and for 2 years (1989-1990) following treatment. Surveys were conducted by spacing personnel approx- imately 6 m apart and traversing the fields in a manner to cover each area as entirely as pos- sible. Number of quail coveys, total birds per covey and the number of rabbits flushed were determined. Efforts were made to watch the direction of flush and subsequent movement of animals to eliminate recounting. Fescue Conversion.—One field served as a fescue-dominated control and the other field was converted from a fescue-dominated veg- etative cover to a gr ass/legume mixture. On 5 March 1988, the treatment field was limed in accordance with soil test recommendations. On 28 March 1988, a prescribed fire was used on the entire treatment field to remove heavy litter and duff from the ground. On 14 April 1988, approximately 50% of the area was sprayed with glyphosphate (Roundup) at a rate of 3.9 liter/ha to kill the vegetation. Portions of the field were not sprayed due to steepness of the slopes. A no-till drill was used to plant a mixture of orchard grass (Dactylis glomerata at 5.8 kg/ha), ladino clover (Trifolium repens at 3.9 kg/ha), red clover (Trifolium pratense at 24 kg/ha) and Korean lespedeza (Lespedeza striata at 3.9 kg/ha) on 26 April 1988. Total costs were recorded and a cost/area rate de- termined. Mosaic pattern strip-mowing was conducted during late July 1988 and late July 1990 on the treatment field to control woody plant en- croachment and create a mosaic of herbaceous and early succession woody cover that would provide more optimum quail and rabbit hab- itat (9, 39). RESULTS AND DIscussION Baseline vegetative sampling, completed in September 1987, showed the 2 fields to be similar in vegetative composition prior to treatment. Both fields were dominated by “KY 31” tall fescue and bluegrass (Poa pratensis). The untreated field also had a fairly high com- ponent of prairie dropseed (Sporobolus asper). A total of 115 plant species was identified from the vegetative sampling during all years of the study. Sixty six plant species were doc- AND RABBITS IN KENTUCKY—Sole 3 umented in the untreated field and 101 in the treated field (Table 1). Forty one (41) spe cies were unique to the treated field and six spe- cies were only found in the control field (Table ip): The fescue conversion to the chosen grass/ legume mixture was completed in April 1988 and can be considered only a partial success. The kill on the fescue was not total, and fescue came back as a codominant cover species in the treated field during the study period. Ger- mination and survival of the planted grass/le- gume mixture was poor due to the severe 1988 drought; orchard grass was essentially lost but pean and Korean lespedeza survived better. Korean lespedeza survived the drought better, providing 10.6% and 6.7% vegetative cover in the treated field for 2 years following treat- ment. Legumes responded well to treatment, resulting in 6 species of legumes with at least 1% total cover on the tested area. No leg- umes documented in the control field provid- ed 1% cover. Conversion was successful at increasing overall plant diversity and the number of dom- inant plant species (Tables 1, 2). The treated field had 27 species of plants providing at least 1.0% cover compared to 10 species on the un- treated field. Likewise, when looking at plant species providing 4.0% or more cover, the treated field had 11 species while the untreat- ed field had 4 species (Table 2). An index to post-treatment quality of plant cover showed the untreated area to have an index of 4 for quail and 10 for rabbit; for an overall rating of 14. The treated area had an index of 27 Her quail and 32 for rabbit; an overall rating of 59 (Table 2). While the percentage of bare ground re- mained nearly constant and averaged 11.2% on the untreated site, bare ground increased from 14.4% pretreatment to 24.4% on the treated field the year immediately following treatment. However, during the 2 years post- treatment the per cent bare gr ound was lower (7.8%) on the treated field than in the untreat- ed field. Ideal quail habitat should have been between 30% and 60% bare ground to provide adequate space for feeding and movement (39). With the Sep Py of the decrease in bare ground in years 2 and 3 following treatment, all the vegetation changes observed on the 4 TRANS. Kentucky ACADEMY OF SCIENCE 56(1—2) TaBLE 1. Plant species identified on fescue converted and control fields at the Kleber WMA, Owen County, Kentucky. Species Acalypha rhomboidea Achillea millefolium Agrimonia pubescens Allium sp. Ambrosia artemisiifolia Andropogon virginicus Antennaria plantaginifolia Apocynum cannabinum Asclepias syriaca Asclepias tuberosa Aster ericoides Aster patens Barbarea sp. Blephilia ciliata Campsis radicans Carex sp. Carya ovata Celtis occidentalis Chrysanthemum leucanthemum Cirsium arvense Cirsium vulgare Coreopsis lanceolata Cornus obliqua Crataegus sp. Croton sp. Dactylis glomerata Danthonia spicata Daucus carota Desmodium paniculatum Dianthus armeria Digitaria sanguinalis Dipsacus sylvestris Elymus virginicus Eragrostis capillaris Erigeron annuus Euonymus americanus Eupatorium serotinum Euphorbia corollata Euphorbia maculata Festuca arundinacea Fragaria virginiana Fraxinus americana Gleditsia triacanthos Gnaphalium obtusifolium Helianthus mollis Houstonia purpurea Hypericum punctatum Ipomoea pandurata Lespedeza procumbens Lespedeza striata Lespedeza virginica Lonicera japonica Ludwigia sp. Lysimachia quadrifolia Maclura pomifera Melilotus alba Melilotus officinalis Monarda fistulosa Osmorhiza sp. Oxalis stricta Common name 3-Seeded Mercury Yarrow Agrimony Wild Onion Ragweed Broomsedge Pussy-toes Indian Hemp Big Milkweek Butterfly Weed Aster Aster Mustard Blephilia Trumpet Vine Sedge Shagbark Hickory Hackberry Ox-eye Daisy Canada Thistle Bull Thistle Wingstem Silky Dogwood Hawthorn Doveweed Orchard Grass Poor Man’s Grass Queen Anne's Lace Tick Trefoil Deptford Pink Crabgrass Teasel Wild Rye Love Grass Daisy Fleabane Strawberry Bush Boneset Flowering Spurge Euphorbia Tall Fescue Wild Strawberry White Ash Honey Locust Gnaphalium Sunflower Houstonia St. John’s Wart Morning Glory Trailing lespedeza Korean lespedeza Virginian Lespedeza Honeysuckle Rattlebox Whorled Loosestrife Osage Orange White Sweet Clover Yellow Sweet Clover Monarda Sweet Sicily Wood Sorrel Untreated 4 KK KK OK Treated ~~ KO Ec co cao ca oo me KK KK KOK KK KOK RR RR RRR | os oo CHANGES IN VEGETATION, QUAILS, AND RABBITS IN KENTUCKY TABLE 1. Continued. Species Common name Untreated Treated Panicum capillare Panic Grass x x Panicum clandestinum Deer Tongue x x Panicum microcarpon Panic Grass i xX Parthenocissus quinquefolia Virginia Creeper x x Paspalum sp. Paspalum x x Physalis sp. Ground Cherry x x Plantago lanceolata Plantain % x Plantago major Plantain x x Poa pratensis Bluegrass x x Potentilla simplex Cinquefoil x = Prunus serotina Black Cherry = x Quercus prinus Chestnut Oak - x Rhus copallinum Winged Sumac = x Rhus glabra Smooth Sumac x x Robinia psuedoacacia Black Locust = x Rosa carolina Carolina Rose Xx Xx Rosa multiflora Multiflora Rose x x Rubus sp. Blackberry X x Rubus flagellaris Dewberry x x Rubus pensilvanicus Raspberry x x Rudbeckia hirta Black-eyed Susan x x Ruellia caroliniensis Wild Petunia x x Rumex acetosella Dock x x Sanicula canadensis Sanicula - x Setaria glauca Foxtail x x Smilax bona-nox Greenbriar = x Solanum carolinense Horse Nettle x x Solidago altissima Field Goldenrod x x Solidago nemoralis Gray Goldenrod - x Spiranthes cernua Ladies Tresses x - Sporobolus asper Prairie Dropseed x x Symphoricarpos orbiculata Coralberry x x Toxicodendron radicans Poison Ivy x x Tridens flavus Greasy Grass x ne Trifolium pratense Red Clover ~ x Trifolium procumbens Yellow Hop Clover x x Trifolium repens White Clover ~ c Ulmus alata Winged Elm = x Unknown mint Sticky Purple Stuff x x Verbascum thaspus Woolly Mullein = x Verbena simplex Verbena = x Verbesina occidentalis Crownbeard = x Vernonia altissima Ironweed x x Viburnum prunifolium Black Haw x x Viburnum rifidulum Black Haw x x Viola sp. Violet x x Vitis vulpina Frosty Grape — Xx Total species 66 101 115 treated field would be considered beneficial to quail and rabbits utilizing the area. The mosaic mowing pattern implemented on the treated field had a positive impact on vegetation important to quail and rabbit. Es- cape cover resulting from the establishment of species such as blackberries, raspberries, dew- berries (Rubus spp.), coralberry (Symphori- carpos orbiculatus), goldenrod (Solidago altis- sima), sweet clover (Melilotus officinalis), and crown beard (Verbesina occidentalis) was de- veloped in desired patterns by mowing the treated field. Herbicide treatment released woody species from the fescue domination. 6 TRANS. KENTUCKY ACADEMY OF SCIENCE 56( 1-2) TABLE 2. Plant species with >4.0% cover found in 1 m? plots on fescue converted and control fields at the Kleber WMA, Owen County, Kentucky indexed to indicate value for food or cover for bobwhite quail and cottontail rabbits. Food value Cover value Species Quail Rabbit Quail Rabbit Quail index Rabbit index Total index Converted field Festuca arundinacea poor poor poor poor 0) 0 0) Poa pratensis poor good fair good 1 4 5 Lespedeza procumbens good good fair fair 3 3 6 Lespedeza striata good good fair fair 3 3 6 Symphoricarpos orbiculata fair good good good 3 4 U Solidago altissima poor poor fair good 1 2 3 Melilotus officinalis good good good good 4 4 8 Tridens flavus fair fair fair fair 2 2 4 Rubus spp. fair fair good good 3 3 6 Ambrosia artemisiifolia good good good good 4 4 8 Panicum capillare good good fair fair 3 3 6 27 32 59 Control field Festuca arundinacea poor poor poor poor 0 0 0 Poa pratensis poor good fair good 1 4 5 Sporobolus asper poor good good good 2 4 6 Solidago altissima poor poor fair good ul 2 is) 4 10 14 Mowing controlled woody invasion and pro- vided escape cover approaching the optimum distribution of within 100 m of other cover types (9, 39). There was a change in utilization of the fields by quail and rabbits following fescue conversion; no use was detected in surveys prior to treatment. The fall following treat- ment, at least 1 covey of quail had become established on the treated field and 1 rabbit was flushed during the survey. The next fall, a covey of birds was again found on the treated area and 3 rabbits were flushed. During the last survey period no quail were found; how- ever, 5 rabbits were observed. It is assumed these animals represent an increase to the lo- cal populations. The numbers should be con- sidered conservative. Each year rabbits were flushed which could not be positively identi- fied as different from a previously flushed rab- bit and were not counted. The flush-survey method has been shown to find approximately 50% of a quail population (41). Therefore, there were likely 2 coveys of quail using the treated field for 2 years immediately following treatment. After the initial year following treatment, habitat quality for quail declined due to the lack of bare ground (39). By the 1990 survey, the vegetation on the treated field may have become too thick for quail utilization: This suggests a need for vegetative disturbance on a 3-year rotation in order to keep ground level vegetation open enough for quail use. No quail or rabbit use was found on the untreated field until the last survey. During that survey a pair of quail were flushed from a multiflora rose thicket in a draw crossing the field. It is hypothesized that these quail were simply using the brushy corridor to cross the area and were not residents of the field. This single replication study cannot make conclusive statements on the value of no-till fescue conversion to rabbit and quail popula- tions. However, the results support the prac- tice of converting fescue to other plant cover types. Native and planted legumes, which are a major food source for quail and rabbits, (9, 12S. 22) 23526, 276 30) 3136. sONtSaee responded very favorably. Bare ground in- creased initially, to allow better movement and feeding by quail. Cost analysis for the fescue conversion and mosaic pattern mowing are shown in Table 3. The costs for this project totaled $182.15/ha for chemicals, no-till equipment rental, and seed for the area treated. About 20 manhours/ ha labor were required to accomplish the task. The costs for seed and no-till drill rental on this project were more than half the total ex- CHANGES IN VEGETATION, QUAILS, AND RABBITS IN KENTUCKY—Sole " TABLE 3. Costs based on 1988 prices. Item Direct $ cost Prescribed fire Herbicide treatment Chemical $81.90/ha 3.9 liters/ha “Roundup” $21/liter Spraying No-till seeding Drill rental $7.75/ha Seed $92.50/ha per mix used on area Mowing Total costs $182.15/ha penses. Due to the extreme drought condi- tions experienced during the 1988 growing season, very little resulted from planting the orchard gr ass/legume mixture. However, the grasses, forbs and legumes released by the burning and spraying alone provided excellent cover and food for bobwhite quail and eastern cottontails. This suggests little need for plant- ing a grass/legume mixture on fescue conver- sion sites where enhancing wildlife habitat is the primary objective. ACKNOWLEDGMENTS Several people were involved in the accom- plishment of this study. Thanks to Doug Mar- shall (KWMA Manager), Jimmy May (District Wildlife Biologist), Tritth District KDFWR Conservation Officers and students from East- ern Kentucky University for assisting in field work, and to Ron Pritchert and Tom Edwards for critical review of the manuscript. LITERATURE CITED 1. Schaaf, L. E., J. D. Sole, T. Edwards, J. Phillips, M. Price, and E. Schneider. 1990. Hunter/harvest survey and deer hunter survey. Kentucky Dept. Fish and Wildl. Re- source. and Urban Res. Inst., Louisville, Kentucky. 2. U.S. Department of Interior. 1989. 1985 National survey of fishing, hunting, and wildlife-associated recrea- tion—Kentucky. U.S. Dept. Interior, Fish and Wildlife Ser., Washington, D.C. 3. Sole, J. D. 1993. Quail and rabbit roadside survey. Cost analysis of a no-till fescue conversion project on a field at Kleber WMA, Owen County, Kentucky. Manhours Tractor fuel used 5 hr/ha 7.5 hr/ha 0.4 ha/liter 7.5 hr/ha (seeding) 0.4 ha/liter 7.5 hr/ha/yr 20 hr/ha (implementation) 7.5 hr/ha/yr (mosaic maintenance mowing) 0.4 ha/liter P-R Proj. W-45-24. Kentucky Dept. of Fish and Wildl. Res., Frankfort. A. Siegel, M. R., M. C. Johnson, D. R. Varney, W. C. Nesmith, R. C. Buckner, L. P. Bush, P. B. Burrus, T. A. Jones, and J. A. Boling. 1984. A fungal endophyte in tall fescue: incidence and dissemination. Phytopathology 74: 932-936. 5. Stoddard, H. L. 1931. The bobwhite quail: its hab- its, preservation and increase. Charles Scribner’s & Sons, New York, New York. 6. Klimstra, W. D. and V. C. Ziccardi. 1963. Night- roosting habitat of bobwhites. J. Wildl. Manage. 27:202— 214. 7. Betsill, C. W., W. L. Matthews, and L. G. Webb. 1979. Cottontail rabbit habitat utilization of a seeded and mechanically maintained power line right-of-way. Proc. Ann. Southeastern Assoc. Fish and Wildl. Agencies 33:20- 24. 8. Sadler, K. C. 1980. Of rabbits and habitat: a long term look. Pennsylvania Game News 51:4—7. 9. Allen, A. W. 1984. Habitat suitability index models: eastern cottontail. U.S. Fish Wildl. Serv. FWS/OBS-82/ 10.66. 10. Stauffer, D. F., G. A. Cline, and M. J. Tonkovich. 1990. Evaluating potential effects of CRP on bobwhite quail in piedmont Virginia. Trans. N. Am. Wildl. and Nat. Res. Conf. 55:57-67. 11. Burger, L. W., E. W. Kurzejeski, T. V. M. R. Ryan. 1990. Structural characteristics of vegetation in CRP fields in northern Missouri and their suitability as bobwhite habitat. Trans. N. Am. Wildl. and Nat. Res. Conf. 55:74-83. 12. Guiliano, W. M. 1990. Food habits, habitat utili- zation, and abundance of eastern cottontail in Kentucky. M.S. Thesis. Eastern Kentucky Univ. Dailey, and 8 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-2) 13. Guiliano, W. M., C. L. Elliott, and J. D. Sole. 1994. Significance of tall fescue in the diet of the eastern cot- tontail. Prairie Naturalist 26:53-60. 14. Lacefield, G. D., J. K. Evans, and R. C. Buckner. 1979. Tall fescue. Univ. of Kentucky, College Ag., Coop. Ext. Serv., Lexington. AGR-59. 15. Bond, H., W. H. Hawk, C. P. Lynch, and C. Jack- son, Jr. 1982. Lower fertility in ewes grazing tall fescue pasture. J. Anim. Sci. 55:44. 16. Heimann, E. D., L. W. Garrett, R. E. Loch, J. S. Morris, and E. H. Pfonder. 1981. Selenium and repro- ductive abnormalities in pregnant pony mares grazing fes- cue pastures. Proc. Equine Nutr. Symp. 7:62-67. 17. Lechtenberg, V. L., W. H. Smith, D. G. Petritz, and K. G. Hawkins. 1975. Performance of cows and calves grazing orchard grass, tall fescue and tall fescue-legume pastures. Beef-Forage Research Day Report. 18. Varney, D. R., M. Ndefru, S. L. Jones, R. New- some, M. R. Siegel, and P. M. Zavos. 1987. The effect of feeding endophyte infected tall fescue seed on reproduc- tive performance in female rats. Comp. Biochem. Physiol. 87C:171-175. 19. Zavos, P. M., B. Salim, J. A. Jackson, D. R. Varney, M. R. Siegel, and R. W. Hemker. 1986. Effect of feeding tall fescue seed infected by endophytic fungus (Acremon- ium coenophialum) on reproductive performance in male rats. Theriogenology 25:281—290. 20. Zavos, P. M., D. R. Varney, M. R. Siegel, R. W. Hemken, J. A. Jackson, and L. P. Bush. 1987a. Effects of feeding endophyte-infected tall fescue seed on the repro- ductive performance in male and female CD-1 mice by combination crosses. Theriogenology 27:541-548. 21. Zavos, P. M., J. A. Jackson, M. R. Siegel, L. P. Bush, and R. W. Hemken. 1987b. Effect of feeding endophyte (Acremonium coenophialum) infected tall fescue seed on reproductive performance in CD-1 mice through contin- uous breeding. Theriogenology 27:549-559. 22. Baldwin, W. P., Jr. and C. O. Handley. 1946. Win- ter food of bobwhite quail in Virginia. J. Wildl. Manage. 10:142-149, 23. Barbour, R. 1951. Observations on the fall and winter food of bob-white quail in Kentucky. J. Wildl. Man- age. 15:108. 24. Bruna, J. F. 1952. Kentucky rabbit investigations. Kentucky Dept. of Fish and Wildl. Res., P-R Proj. W-26- F. Frankfort, Kentucky. 25. Burger, L. W., E. W. Kurzejeski, T. V. Dailey, and M. R. Ryan. 1992. Relative invertebrate abundance and biomass in Conservation Reserve Program plantings in northern Missouri. Proc. Nat. Bobwhite Quail Symposium Sele, 26. Casey, W. H. 1965. Some speculations on the min- imum habitat requirements of bobwhite quail. Proc. Ann. Conf. Southeast. Assoc. Game and Fish Comm. 19:30-39. 27. Dalke, P. D. and P. R. Sime. 1941. Food habits of the eastern and New England Cottontail. J. Wildl. Man- age. 5:216-228. 28. Durell, J. 1952. Farm game habitat restoration in Kentucky. Proc. Annu. Conf. Southeast. Assoc. Fish Wildl. Agencies 6:559-566. 29. Dusi, J. L. 1952. The food habits of several pop- ulations of cottontail rabbits in Ohio. J. Wildl. Manag. 16: 180-186. 30. Henshaw, M. 1977. Major quail foods: Kentucky. KDFWR in-house report. Kentucky Dept. Fish and Wildl. Res., Frankfort, Kentucky. 31. Hurst, G. A. 1972. Insects and bobwhite quail brood habitat management. Proc. Nat. Bobwhite Quail Symp. 1:65-82. 32. Johnsgard, P. A. 1973. Grouse and quails of North America. Univ. Nebraska Press, Lincoln, Nebraska. 33. Klimstra, W. D. 1982. Bobwhite quail and chang- ing land use. Proc. Nat. Bobwhite Quail Symp. 2:1-5. 34. Roseberry, J. L. and W. D. Klimstra. 1984. Popu- lation ecology of the bobwhite. Southern Illinois Univ. Press, Carbondale, Illinois. 35. Rosene, W. R., Jr. 1969. The bobwhite quail: its life and management. Rutgers Univ. Press, New Bruns- wick, New Jersey. 36. Rosene, W. R., Jr. and J. D. Freeman. 1988. A guide to and culture of flowering plants and their seed important to bobwhite quail. Morris Commun. Corp., Au- gusta, Georgia. 37. Robel, R. J. 1972. Body fat content of bobwhites in relation to food plantings in Kansas. Proc. Nat. Bob- white Quail Symp. 1:139-149. 38. Simpson, R. 1989. Bobwhite brood habitat in the southeast—farm chemicals and wildlife—can they co-ex- ist? Proc. Quail Unlimited National Quail Seminar 1:95— 96. 39. Schroeder, R. L. 1985. Habitat suitability index models: northern bobwhite. U.S. Fish Wildl. Serv. Biol. Rep. 82 (10.104). 40. Trippensee, R. E. 1948. Wildlife management— upland game and general principles. Vol. I. McGraw-Hill Book Co., New York, New York. 479 pp. 41. Umber, R. W,, C. Pregler, and J. H. Eve. 1979. The bobwhite quail in Oklahoma. Oklahoma Dept. Wildl. Conserv., Stillwater. 42. Dimmick, R. W,, F. E. Kellogg, and G. L. Doster. 1982. The Lincoln Index and flush census as population estimates for bobwhites. Proc. Nat. Bobwhite Quail Symp. 2:13-18. 43. Bookhout, T. A. 1958. The availability of plant seeds to bobwhite quail in southern Illinois. Ecology 39: 671-681. 44, Dickson, J. G. 1971. Food habits of the bobwhite quail in the Georgia piedmont. Proc. Ann. Conf. South- east. Assoc. Game and Fish Comm. 25:281—285. Trans. Ky. Acad. Sci., 56(1—2), 1995, 9-14 Human Sex Ratio and Factors Influencing Family Size in Hunan, China ELMER Gray,* VALINA K. Hurt,” AND Ju Y. Wue “Department of Agriculture, Western Kentucky University, Bowling Green, Kentucky 42101-3576 Division of Science and Related Technologies, Hazard Community College, Hazard, Kentucky 41701 ‘Hunan Provincial Family Planning Research Institute, Hunan, People’s Republic of China ABSTRACT In 1990, sex ratio and family size data were obtained on 1,002 families of university students from Hunan Province, China. Each student respondent provided data on the parental, present, and projected generations. Secondary sex ratios (males per 100 females) were 114.6, 101.3 and 110.0 and average numbers of children were 4.32, 3.36, and 1.65 for the parental, present, and projected generations, respectively. Disparity between observed and expected combinations of sexes of children among families of the parental and present gen- erations indicated a preference for both sexes and for male children. No significant correlations were found between sexes of children within families. Student respondents expressed strong preferences for family size and for specific combinations and permutations of sexes of children. The most desired family consisted of 2 children including both sexes with the first-born being male. For the projected generation, the resulting sex ratio (110) would not create a serious imbalance of the sexes and the resulting average family size (1.65 children) would be supportive of China’s population control effort. INTRODUCTION The increasing human population continues to be a global threat and a national concern for many countries. The human population is a function of the number of families and fam- ily size. Control efforts are largely directed to- ward family size. Reduction in family size ap- pears to be the result of deliberate family planning. Greater understanding of the human sex ra- tio and of the factors that influence family size would contribute to family planning. In some populations, evidence has been found that the combinations of sexes of existing children in- fluenced the parents’ decision to have addi- tional children. In some cultures fewer fami- lies had additional children after both sexes were represented. In other cultures, the like- lihood of additional children was influenced by the number of sons among existing children. Survey results from diverse cultures have re- vealed that individuals have definite prefer- ences for family sizes consisting of specific combinations and permutations of sexes of children. The China population is estimated to be ap- proximately 1.1 billion and to be increasing at the rate of 1.4 percent annually, a rate that 3) would double the population in 49 years (14). Approximately 27 per cent of the population is under the age of 15 years. As the most pop- ulous country in the world, China’s population control will have a substantial impact on the world population (22). Although the Chinese Government has given major emphasis to fam- ily planning since the early 1970s, the State Council’s call for “only one child per couple” was issued in 1980 (21). It was acknowledged that implementation of the one-child-per-cou- ple measure may result in social problems; however, that control would enable China to keep its population within a 1.2 billion limit. The objective of the present investigation was to extend the basic studies of the human sex ratio and of the influence of combinations of sexes of children on family size to a popu- lation which has reached unequaled size and which has been subjected to unparalleled con- trol measures. POPULATION AND PROCEDURES In 1990, data were obtained on sex of chil- dren and family size for 1,002 university stu- dents from Hunan Province, China. The stu- dents were enrolled at one of 3 educational institutions—Hunan Medical University, Changsha Medical School, or Hunan Teachers 10 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) University. Each student completed a data form providing information on the parental, present, and projected generations. For the parental generation, information was collected on the number of children by sex for both the maternal and paternal par- ents. These data were used to calculate aver- age family size and sex ratio, and for compar- ison of the observed and expected composi- tion of sexes of children within various family sizes. For the present ¢ generation, information was obtained on sex of children by order of birth, permitting exploration of associations between sexes of children and the effects of combinations and permutations of sexes of children on family size. Linear correlation was used to test associations between sexes and chi-square for a 2 X 2 contingency table was used to test the independence of combinations of sexes and family size (17). For the projected generation, each student respondent indicated the desired number of children and the pre- ferred combination and permutation of sexes. Using these data, average desired family size and the resulting sex ratio were calculated. RESULTS Binomial distribution.—The number of families of sizes | through 4 and their various combinations of sexes for the parental and present generations were determined. Ex- pected frequencies of the various combina- tions of sexes were calculated using the ob- served sex ratio for each generation in the bi- nomial expansion. In the parental generation, the only significant divergence between ob- served and expected combinations of sexes oc- curred in 2-children families. The combina- tion of 1 male and 1 female was detected more frequently than expected. In the present gen- eration, the only significant difference be- tween observed and expected combinations occurred in three-children families. The com- bination of 2 males and 1 female appeared more frequently than expected. Disparity be- tween observed and expected distributions of combinations of sexes within various family sizes was not found in other populations (8 WO), eh, IS), Correlations between Sexes.—Correlation coefficients were calculated between sexes of consecutive and nonconsecutive children with- in families of the present generation (Table 1). > TaBLE 1. Correlation coefficients between sexes of chil- dren of different births in the present generation. Family size and birth order Number Correlation All families Birth 1 vs. birth 2 778 —0.022 (ns)# Birth 1 vs. birth 3 585 —().039 (ns) Birth 2 vs. birth 3 585 —(0.052 (ns) Families of two children Birth 1 vs. birth 2 193 —0.064 (ns) Families of three children Birth 1 vs. birth 2 291 —0.051 (ns) Birth 1 vs. birth 3 291 —0.111 (ns) Birth 2 vs. birth 3 291 —0.112 (ns) “ns nonsignificant at the 0.05 level of probability. Those associations were explored among the first three births for all families and for fami- lies consisting of only two or three children. All of the coefficients were low in magnitude, negative in sign, and nonsignificant. Sexes of successive children have been found to be positively correlated in some subsets of the population (4, 8, 10, 15), but not in others (5, iS) Sex Ratios and Average Family Sizes.—Sec- ondary sex ratios G@qualess per 100 females) were 114.6 and 101.3 for the parental and present generations, respectively. If the true ratio is greater than 100, as indicated by the parental ratio, the ratio for the present generation was biased downward by the inclusion of an equal number of female and male respondents. These sex ratios are within the range of those reported for other subsets of the human pop- ulation. Examples of ratios reported for other populations include: 101 for Black Americans (13), 106 for White Americans (18), 107 for Brazilians (8), 107 for Japanese (10), and 106 for Libyans (2). Sex ratios by order of birth were 91.4, 95.5, 117.5, and 98.6, respectively, for the first 4 births in the present generation (Table 2). The ratio of 117.5 for the third birth was significantly different from the others, but did not differ significantly from the overall ra- tio of 101.3 for the present generation. No bi- ological explanation is evident for the aberrant ratio for the third birth. Average numbers of children were 4.32 for the parental and 3.36 for the present genera- tions. These averages were lower than corre- sponding values reported for Brazilian (8) and Libyan (2) populations. However, they were HUuMAN Sex Ratios IN CutnA—Gray, Hurt, and Wu 1] TABLE 2 birth in the present generation. Sex ratios (males per 100 females) Family Birth oe ee = Combined order Stopped Continued for birth l Number 20 778 798 Sex ratio 222, 90.7 91.4 2 Number 193 585 778 Sex ratio 114.4 89.9 95.5 3 Number 291 294 585 Sex ratio 136.6 101.4 MIT 3 4 Number 159 135 294 Sex ratio 144.6 62.6 98.6 approximately equal to average numbers of children (4.56 and 3.32) reported for a United States population (15). The decrease in aver- age number of children (4.32 to 3.36) from the parental to the present generations in the Chi- na population is considerably less than the cor- responding decrease (5.16 to 2.43) reported for a Japanese population (10). Combinations of Sexes and Family Size.— Composition of sexes of the first 2 children had no significant effect on family size (Table 3). However, the proportions of families stop- ping with 3 and 4 children were influenced by the sexes of existing children. For both 3 and 4 children families, the percentages of families that ceased to have additional children were higher when both sexes were present than when only 1 sex was present. Presence of both sexes in the first 2 children has been associ- ated with smaller families in Britain (19) and in the United States (7, 13). That association was not found in another United States study (3), in Brazil (8, 20), in Japan (10), or in Libya (2). The influence of the composition of sexes of children on family size in the present study is similar to that reported for Black and Ap- palachian populations in the United States (13). In those populations, the composition of sexes of the first 2 children had no influence on family size; however, the presence of both sexes in the first 3 or 4 children reduced the likelihood of additional children. In addition to the preference for both sexes, there was preference for a prevalence of male children. Higher percentages of families ceased having children when existing children were all males than when all were females and when there were at least 2 male children among the first 3 or 4 children (Table 3). Also, by order of TABLE 3. children on family size in the present generation. Influence of composition of sexes of existing Family Family stopped increased Sex composition No % No % First child Outcomes (ns)" m 11 2.9 OR ial f 9 NO) 408 97.8 Total 20 hay Tits) STS First two children All combinations and permutations (ns) mm 46 26.1 SO) SLY. mf 46 23.8 A eeeioee, fin Bye BAS) 7B ff AU AL I5) LOL 7825 Total 1K ae: Sr to tos a fo 2 First three children All combinations and permutations (*) mmm 29 46.0 34 54.0 mmf 40 59.7 27 ~=—40.3 mfm AGS 5353(6) 36 44.4 fmm 49 59.8 Bon 402 mff 98 42.4 88) 56 finf Sl 27/0) Bi axshi0) ffm 45 50.0 45 50.0 fff AR OATS 46 65.7 Total 291 49.7 294 50.3 First four children All combinations (**) Four males 8 50.0 8 50.0 Three males, one female 42 63.6 94 36.4 Two males, two females 66 62.3 AN), Sif 7f One male, three females 37 44.0 47 56.0 Four females Wee Gea Total 159 54.1 135 45.9 “* and **, P < 0.05 and 0.01, respectively, that increases in family size are independent of sexes of existing children; ns, P > 0.05 that increases in family size are independent of sexes of existing children the sex ratio for the last birth within families (Table 2) indicated that families more often stopped having children after the birth of a male than after the birth of a female child. Park (16) reported a male preference for Ko- rean families. The percentages of families pro- ducing a fourth child were 75.8, 80.6, 83.9, and 87.0, respectively, when the numbers of males in the first 3 children were 3, 2, 1, and 0. Gray (6) found evidence of the male pref- erence among families of a United States pop- ulation. Projected Generation Data for desired family size, combinations, and permutations of sexes of children are pre- 12 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) TaBLE 4. Desired family size, combination, and permu- tation of sexes of children. Respondents Permuta- Respondents No. Combination ———————————___ tion children of sexes No. % of sexes No. %G 0 68 8.5 1 lm 123 emia: lf OES Total 215 269 2) 2m 13 1.6 Im, lf 464 58.1 mf 306 = 338.3 fm 158 19.8 of 2 0.2 Total 479 60.0 3 3m 0 0.0 2m, If 15 1.9 mmf 8 1.0 mfm a 0.9 fmm 0) 0.0 1m, 2f 4 0.5 mff 2 0.2 fmf 0 0.0 ffm 2 0.2 3f 0 0.0 Total Ty ee >3 17 2.1 sented in Table 4. Approximately 8.5 per cent of the respondents wanted no children. In similar studies of other subsets of the human population, the percentages of respondents wanting no children ranged from a low of 0.0 for Nigeria (11) to a high of approximately 8.4 for a United States population (6). In the pres- ent study, the most desired family size (60% of respondents) consisted of two chiidren which was also the most preferred family size in the United States (6, 12), in Japan (10), and in Brazil (9). Only 2.4% of the Chinese re- spondents wanted three children; only 2.1% wanted more than three children. The result- ing average family size of 1.65 children was lower than that reported for other subsets of the population—United States 2.60 (6) and 2.46 (12), Brazil 2.76 (9), Japan 2.35 (10), Ni- geria 4.88 (11) and in Libya 4.40 (1). The Chinese respondents expressed strong preference for both sexes of children. For ex- ample, 58.1% of the respondents wanted 1 fe- male and | male; whereas, 1.8% wanted either 2 females, or 2 males. In addition to the pref- erence for both sexes, there was a preference for male children as evidenced by greater fre- quencies of 1 male over 1 female in 1-child families, of 2 males over 2 females in 2-chil- dren families, and of 2 males and 1 female TABLE 5. Sex ratios, by order of birth, resulting from desired family size, combination, and permutation of sexes of children. Sex ratio by order of birth (males: 100 females) Number of Overall Desired family size _ respondents 1 2 3 births 1 215 134 134 2 479 199 56 105 3 19 850 89 90 148 >3 17 112 183 112 112 Overall sizes 730 179 59 100 4110 over | male and 2 females in 3-children fam- ilies. Approximately 64% of the respondents wanted the first-born child to be male. These preferences for both sexes and for the first child to be male are compatible with results from studies of other populations (1, 6, 11, 9, 12). Since few Chinese respondents wanted 3 or more children, it was not possible to ex- plore the preference for alternation of sexes (mfm, mfmf) that has characterized other sub- sets of the population. Sex ratios (males per 100 females) resulting from desired family sizes, combinations, and permutations of sexes of children by desired family size and order of birth are given in Ta- ble 5. Preference for the first-born to be male was again evidenced by the ratio of 179 for the first birth overall family sizes. The sex ratio (110) overall family sizes and birth orders was within the range of the observed ratios for the parental (114.6) and present (101.3) genera- tions. The 110 ratio would result in more equal proportions of females and males for the Chinese population than has been reported for other populations. Ratios for desired fam- ilies in other populations have ranged from a low of 121.3 for Brazil (9) to a high of 167.0 for Nigeria (11). DISCUSSION The China population was selected because of its unequaled size and its recent subjection to intensive family planning measures. The population control efforts have been directed toward limiting the number of children per family. Utilization of college and university students did not result in a totally represen- tative sample of the China population; how- ever, the study did include the parental gen- eration of the college students as well. Other HUMAN SEx Ratios IN Cuina—Gray, Hurt, and Wu 13 reported studies have, likewise, utilized col- lege students and their families. The China population had some similarities and some differences when compared with other populations. Similarities with certain other populations included: magnitude of ob- served sex ratios, lack of significant correla- tions between sexes of successive children within families, average number of children per family, magnitude of decrease in family size between parental and present genera- tions, influence of both sexes and male chil- dren on family size, and preferences for 2- children families consisting of both sexes with the first-born being male. The China population differed from others in that the observed and expected frequencies of combinations of sexes within families were not always equal. Discrepancies were ob- served in both the parental and present gen- erations. Those departures were indicative of parental preferences for both sexes and for male children. Although there was evidence of preference for male children in the present generation and of strong preference for the first-born to be male in the projected gener- ation, the resulting sex ratio (110) for ehe pro- jected generation is closer to equal numbers of males and females than that reported for any other population. In fact, the 110 ratio for the projected generation is lower than the 114.6 observed ratio for the parental genera- tion. Closeness of the sex ratio for the desired families may be a result of China’s recent em- phasis on the fact that ancestral lines are con- tinued through female as well as male children (21). If the knowledge of human reproduction reaches the level where parents can have their choice of sexes, the resulting imbalance of sex- es should be less in the China population than in other populations where the sex ratio for desired families is higher. For example, the re- sulting ratio for a projected generation of Ni- gerian families was 167 males per 100 females (lel), In the present study, respondents wanted an average of 1.65 children per family, which is the smallest desired family size of any of the populations that have been studied. However, the 1.65 children per family is far greater than the one child per family oncom by Chi- na’s present population policy (22). Approxi- mately 27% of respondents wanted a 1-child family. Wenruo (21) conceded that the fertility rate for the end of this century is still expecte d to be approximately 1.5 children per couple. The China Government initiated vigorous population planning programs in the early 1970s and has greatly intensified those pro- grams since 1980. In the present study, the parental generation was born around 1950; the present generation of college students was born around 1970. Thus, both generations preceded the intensive family planning im- pact. It would be enlightening to study the sex ratio of the smaller families of the 1980s. ACKNOWLEDGMENT We are grateful to Yanchu Fang and Xinmin Yue for their role in facilitating the study. LITERATURE CITED 1. Abdulla, A. S., M. A. F. Al-Rubeai, and E. Gray. 1984. Desired family size and sex of children in Libya. J. Hered. 75:76-78. 2. Al-Rubeai, M. A. F., A. S. Abdulla, and E. Gray. 1983. Analysis of the human sex ratio: factors influencing family size in Libya. J. Hered. 74:39-42. 3. Ayala, F. J. and C. T. Falk. 1971. and family size. J. Hered. 62:57-59. 4. Edwards, A. W. F. 1961. A factorial analysis of sex ratio data. Ann. Hum. Genet. 25:117-121. 5. Edwards, A. W. F. and M. Fraccaro. 1960. Distri- bution and sequences of sexes in a selected sample of Swedish families. Ann. Hum. Genet. 24:245-252. 6. Gray, E. 1982. Transgeneration analyses of the hu- man sex ratio. J. Hered. 73:123-127. 7. Gray, E. 1972. Influence of sex of first two children on family size. J. Hered. 63:91-92. 8. Gray, E. and J. Bortolozzi. 1977. Studies of the hu- man sex ratio and factors influencing family size in Bo- tucatu, Brazil. J. Hered. 68:241-244. 9. Gray, E., J. Bortolozzi, and V. K. Hurt. 1979. De- sired family size and sex of children in Botucatu, Brazil. J. Hered. 70:67-69. 10. Gray, E., D. Duckworth, and Y. Nakajima. 1980. The human sex ratio and factors influencing family size in Japan. J. Hered. 71:411-415. 11. Gray, E., V. K. Hurt, and S. O. Oyewole. 1983. Desired family size and sex of children in Nigeria. ]. Her- ed. 74:204—206. 12. Gray, E. and D. K. Morgan. 1976. Desired family size and sex of children. J. Hered. 67:319-321. 13. Gray, E. and N. M. 1974. Influence of combinations of sexes of children on family size. J. Hered. 65:169-174. 14. Haub, C., M. M. Kent, and M. Yanagishita. 1990. 1990 World population data sheet. Population Reference Washington, D.C. Sex of children Morrison. Bureau, Inc., 14 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) 15. Loyd, R. C. and E. Gray. 1969. A statistical study of the human sex ratio. J. Hered. 60:329-331. 16. Park, C. B. 1978. The fourth Korean child: the effect of son preference on subsequent fertility. J. Biosco. Sci. 10:95—106. 17. Steel, R. G. D. and J. H. Torrie. 1980. Principles and procedures of statistics, 2nd ed. McGraw-Hill Book Co., New York. 18. Stern, C. 1960. Principles of human genetics, 2nd ed. W. H. Freeman, San Francisco. 19. Thomas, M. H. 1951. Sex patterns and size of fam- ily. Br. Med. J. 1:733-734. 20. Varella-Garcia, M. 1977. Children’s sex in a Bra- zilian population of Arabian origin. Naturalia 3:45-48. 21. Wenruo, H. 1981. Population policy. In L. Zheng and J. Song (eds.) China’s population: problems and pros- pects. New World Press, Beijing. 22. Zheng, L. 1981. Population planning and demo- graphic theory. In L. Zheng and J. Song (eds.) China’s population: problems and prospects. New World Press, Beijing. Trans. Ky. Acad. Sci., 56(1—2), 1995, 15-21 Decomposition of Nerve Gas and Mustard Gas Analogs Using Nicotine and Ultrasound JOHN L. MEISENHEIMER, LAWRENCE R. MILLER, AND WILLIAM D. SCHULZ Department of Chemistry, Eastern Kentucky University, Richmond, Kentucky 40475 ABSTRACT There are appreciable amounts of mustard gas (HD) and the nerve gases GB and VX stored in Madison County, Kentucky. This research employed nicotine and ultrasound in a study of the chemical decomposition of chemical-physical analogs of these toxic agents. The insecticide malathion was selected as an analog of VX due to the presence of phosphorus-sulfur bonds in both compounds and similar physical properties. 2- Chloroethyl ethyl sulfide was used as the analog of mustard gas alone was found to effect a complete decomposition of these compounds, but the reaction with 2- 5 (HD), bis(2-chloroethyl) sulfide. Nicotine chloroethyl ethyl sulfide was slow. The application of ultrasound to the nicotine-analog mixtures induced a substantial increase in the rate of analog reaction, thus indicating this method has the potential for practical, “closed- loop” demilitarization of the corresponding warfare agents. INTRODUCTION There are 8 locations about the continental United States, and one at Johnston Island, where some 25,000 tons of chemical warfare agents are stored (1). These agents need to be Ae ciarized for two reasons, (1) to be in compliance with international treaties on de- struction of these agents, and (2) because the containers for these agents are gradually cor- roding and present a long-term hazard to per- sons living or working in the vicinity of the storage areas. The second reason is of particular concern to the citizens of central Kentucky, and es- pecially those in proximity to the Lexington- Blue Grass Army Depot, where the nerve gas- es GB (I) and vx (II) are stored as well as the vesicant agent mustard gas or HD (III). ) fo) | | CH3-P-F CH; - P- SCH,CH,N(CH(CH3))2 OCH(CH3)> OCH,CH, I (GB) II (VX) CICH)CH)SCH,CH,CI Ill (HD) Some of these agents are contained rockets that also hold both explosive bursters and propellant chemicals that make them doubly dangerous as they age. Safe destruc- tion and/or disposal of hese agents is com- plicated by the fact that the storage and pro- 15 posed disposal sites are close (2-3 miles) to civilian housing, businesses, and schools. This proximity leaves absolutely no margin for hu- man error or mechanical accident or mal- function in demilitarization, all possibilities in agent destruction by the army-favored incin- eration process. The proposed incineration of the chemical agents in Kentucky, and at several other stor- age sites around the country, has been op- posed by citizen’s groups for a variety of rea- sons: (1) There is concern because the incinera- tion process requires an “open-loop” system. There is a finite possibility that due to error or accident a major emission of agent could escape from the incinerator stack. Residents of this area believe there would be so many unprotected people close to the incinerator that the possibility of adequate warning and safe evacuation is, arguably, impossible. Such a disaster would thus result in the loss of many lives and the magnitude of the tragedy would be incalculable. (2) Minor amounts of agent might persist in stack effluent even though the health con- sequences of this continued exposure to trace amounts of chemical warfare agent have never been adequately studied. (3) There is a logical concern that after the disposal of the chemical warfare agents no ef- fort would be made to dismantle the inciner- ator. Rather, such an expensive facility would be converted into a general toxic waste incin- 16 TraANs. KENTUCKY ACADEMY OF SCIENCE 56(1—2) erator and be perpetually used to burn com- mercial as well as government toxic wastes. It would serve as a magnet for the transportation of toxic wastes from all of the Eastern United States into central Kentucky. (4) The incineration of mustard gas (HD) has the potential of forming small amounts of the very toxic and persistent 2,3,7,8-tetra- chlorodibenzodioxane (TCDD or “dioxin’”) and other polychlorinated dibenzodioxanes. In animal tests, TCDD is found to be embry- otoxic, teratogenic and perhaps carcinogenic (2, 3). In fact, the organizations Physicians for Social Responsibility and the Environmental Defense Fund have jointly released recom- mendations for a new national policy to re- duce dioxin pollution. These recommenda- tions include a requirement that all incinera- tors operate under conditions that produce virtually no dioxins (4). The general reaction for TCDD production during incineration is shown below: Heat CICH,CH,SCH,CH,C1 —+» CO,+ H,0+ HCl HD O2 + Oxides of sulfur and other oxides DISCUSSION The army has stated its intention to use a demilitarization method that will decompose all of the different types of warfare agents by the same process. Their position is that incin- eration is the only current process that meets that criterion. In part, the argument has some validity. In the past, some GB agent has been satisfactorily demilitarized by using aqueous hydroxide or hypochlorite solutions. However, unlike GB, VX and mustard gas (HD) are not readily or completely soluble in water (5, 6). Therefore, it is difficult to attain complete de- composition of these agents at low tempera- ture—low pressure in aqueous solutions by hy- drolysis (7). We propose that since all of these agents have a leaving group subject to removal by nu- cleophilic attack, all should be able to be de- composed by simple mixing with an effective nucleophile in which they are soluble. We fur- ther suggest that the natural alkaloid nicotine possesses these solubility-nucleophilic prop- erties, thus presenting a novel approach to de- militarization. In addition, we have studied the use of sonochemistry as a method to enhance the decomposition of these agents. Below are listed the expected reaction of each of the agents with nicotine: MUSTARD GAS (HD) + NICOTINE CICH2CH2SCH2CH2C1 = + HD © CHs ue OSG a N a a N In addition, there are some less likely by- products where the pyridine moiety rather than the N-methylpyrrolidine moiety of nico- tine displaces one or two chlorines. The re- action that follows shows a one-chlorine dis- placement by that process: CH, CICH,CH)SCH,CH>Cl + ti HD > a [© NICOTINE es N | N CH3CH,0 @ + (eit and small amounts of: I x CH; = 1D ae HSCH,CH2N(CH(CH3)>)2 CH,CH,O ae N CHY/ Due to the extreme toxicity and unavail- ability of the warfare agents, it has not been possible, to date, for us to perform experi- ments on the agents themselves. Rather, it has been necessary to use commercially available compounds that are reasonable chemical an- alogs for this research. The insecticide mala- thion was selected to serve as the analog of VX since it has a similar phosphorus-sulfur bond, similarly sized substituents on the phos- phorus atom, and similar solubility properties. Its chemical reactivity is sufficiently close to that of VX to cause it, and especially its oxi- dized metabolite malaoxon, to possess some anticholinesterase activity (8). The equations for the expected decomposition of malathion by nicotine are given below: AND MustarpD Gases—Meisenheimer et al. Vy MALATHION + NICOTINE CHO F ‘P—SCHCOOCG,H; + C205 > \ccooca, CH; $ CHOI +! = SPN CH,O _ Plus small amounts of this product: " SCHCOOC3Hs CH,COOC,H, 2-Chloroethyl ethyl sulfide was the logical choice to employ as the experimental analog of mustard gas (HD or bis(2-chloro- ethyl)sulfide). The sensitive 2-chloroethyl sul- fide functional group is the same in both com- pounds, just one in the analog and 2 in mus- tard gas, and they have obvious similarities of size and solubility. The expected nucleophilic displacement reactions of nicotine and 2-chlo- roethyl ethyl sulfide are shown below: 2-CHLOROETHYL ETHYL SULFIDE + NICOTINE CICH2CH2SCH2CH3 at 2 : CH,CH,SCH,CH,-N Oh ta Plus small amounts of the isomer below: + CH,CH,SCH,CH,-N C) 18 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) MATERIALS, INSTRUMENTATION, AND METHODS Reagents: Benzene—The benzene was freshly distilled prior to each use (Fisher, #B-245, ACS cer- tified, b.p. 80°C). 2-Chloroethyl ethyl sulfide—This compound was used as purchased without further pu- rification (Aldrich Chemical Co., #24,264-0, Assay 98% purity, b.p. 156-157°C).* Malathion—This compound was used as pur- chased without further purification (Chem Service, #PS-86, Assay 98% purity, b.p. 5 G= ai Ges Nicotine—This compound was used as pur- chased without further purification (EM Science, #NX0370-1, Assay 98% purity, b.p. IQS nC) es Instrumentation: Gas Chromatograph/Mass Spectrometer— Hewlett Packard 5890 GC and 5970 MS with an automatic sampler 7673A. Column—J & W Inc., DB-5, 50 m X 0.20 mm ID, 0.33 micron polysiloxane film. Sonochemical Reactor—Ace Glass Incorporat- ed #9830 Sonochemical Reaction Assembly with #9818 microtip. Experimental Procedure: All reactions were carried out without heat- ing, except for any heat increase caused by the sonochemical probe. The reactions without sonochemistry were carried out at room tem- perature, 20—22°C. The reactions with sono- chemistry reached 40-41.5°C. This tempera- ture differential accounts for only a portion of the observed rate increase with sonochemistry. Current theory on sonochemical effects holds that as transient, minute cavitation bubbles at the surface of the probe collapse they gener- ate enormous local pressures and tempera- tures. Even though the microbubble collapse may generate temperatures well over 1,000°K, the macroscopic temperature change is only some 15—20° (9). In all sonochemical procedures the probe was programmed to alternately pulse for 2 seconds and rest for two seconds. In all experiments, 5.0 microliter samples * Gave a single peak upon GC analysis. were removed for analysis, diluted in 6.0 ml of benzene, and the concentration of analog determined by GC/MS. The small sample size caused some slight, but acceptable scatter of data points. Each reaction was monitored until no trace of the analog could be detected. Separation of components was carried out for malathion at an initial column temperature of 150°C, increased 10°C/min. to a maximum of 280°C. For 2-chloroethyl ethyl sulfide, ini- tial column temperature was 100°C, increased by 10°C/min. to a final temperature of 230°C. Malathion With Nicotine.—0.330 grams (0.001 mol) of malathion was mixed with 0.324 grams (0.002 mol) of nicotine in a 3.0 ml re- action vial fitted with a water-cooled condens- er. The reaction was stirred continuously with a magnetic-spin vane except for sample re- moval periods. Malathion With Nicotine Plus Sonochemis- try.—3.301 grams (0.01 mol) of malathion was mixed with 3.245 grams (0.02 mol) of nicotine in a cylindrical reaction vessel, open at the top with a sonochemical probe inserted. 2-Chloroethyl Ethyl Sulfide With Nico- tine.—0.249 grams (0.002 mol) of 2-chloro- ethyl ethyl sulfide was mixed with 0.649 grams (0.004 mol) of nicotine in a 3.0 ml reaction vial fitted with a water-cooled condenser. The reaction was stirred continuously with a mag- netic-spin vane except for sample removal pe- riods. 2-chloroethyl Ethyl Sulfide With Nicotine Plus Sonochemistry.—1.246 grams (0.01 mol) of 2-chloroethyl ethyl sulfide was mixed with 3.24 grams (0.02 mol) of nicotine in a cylin- drical reaction vessel, open at the top with a sonochemical probe inserted. EXPERIMENTAL RESULTS Malathion: The experimental results for the reaction of malathion and nicotine, with and without the sonochemical probe, are summarized in Fig- ure 1. Without sonochemistry the 0.33 gram sample (0.001 mol) of malathion was com- pletely decomposed in 600 minutes, with a re- action half-life of 99 minutes. The decay rate was exponential, with y = 113.9 X 10°°* in the best-fit curve. Using the sonochemical reactor, 3.30 grams (0.01 mol) of malathion was decomposed in 42 DECOMPOSITION OF NERVE AND MusTARD Gases—Meisenheimer et al. 19 : < = 2 a iS} E - 2 = z iS} [4 [ea a a REACTION WITH SONOCHEMISTRY ° REACTION WITHOUT SONOCHEMISTRY Fic. 1. Malathion + nicotine. M—reaction with sono- chemistry; © —reaction without sonochemistry. minutes, the reaction half-life dropping to only 9.6 minutes. Again, there was an exponential decay rate for the malathion. The best-fit curve had y = 176.7 X 107°%68x, Even though the reaction was carried out in an open vessel, there was no measurable loss of reactants by evaporation during the reac- tion. 2-Chloroethyl ethyl sulfide: The experimental results for the reaction of 2-chloroethyl ethyl sulfide are displayed on the graph in Figure 2. The decomposition of 0.249 grams (0.002 mol) of this compound by nico- tine alone was quite slow, taking 290 hours for completion. The reaction half-life was approx- imately 150 hours. It is obvious from Figure 2 that the decomposition rate of this com- pound is not exponential like that of mala- thion. In fact, the best-fit curve is a second- order polynomial type that has the sign of the change in slope with time opposite to that of the malathion curve. The 2-chloroethy! ethyl sulfide reaction with nicotine begins slowly, suggesting an induction period. This is prob- ably due to a solvent effect because as the re- actants move toward the transition state there must be forming charges. Since both reactants are neutral, either an S.1 or an S,2 mecha- nism requires charge formation that is not ef- fectively stabilized by the neutral mixture. As the reaction proceeds and ions are produced the reaction environment is more amenable to the forming charges in the transition state. The application of a sonochemical probe to this reaction effected a substantial rate in- crease. The 1.246 gram (0.10 mol) sample of 2-chloroethyl ethyl sulfide was completely de- composed in 41 hours compared to 290 hours without sonochemistry. The reaction half-life was reduced to 15.5 hours in comparison to the 150 hours required without sonoc hemistry. There was no loss of reactants from the open reaction vessel by evaporation. CONCLUSIONS This work demonstrates that analogs of VX nerve gas and mustard gas (HD) can be com- pletely decomposed chemically using nicotine as both reactant and reaction solvent. Also, the experimental results demonstrate that using a sonochemical probe affords a great increase in reaction rate. Additional rate increases would be readily effected by lengthening the time of the sonochemical pulse and increasing the re- action temperature. Further studies to opti- mize solvent polarity should provide even greater rates, especially in the case of mustard gas (HD) decomposition. One can conclude that with the above mentioned enhancements in method and with additional research the use of nicotine for the demilitarization of the aforementioned warfare agents could be a vi- able process. The process does have several features and advantages that address the major concerns of the residents of central Kentucky and the rec- ommendations of the Kentucky Citizens Ad- visory Commission, appointed by Governor Brereton Jones, with respect to demilitariza- tion of the chemical agent stockpile. (1) It would permit a “closed-loop” system so no gases could escape into the atmosphere. (2) The demilitarization reactions could be carried out at low temperature-low pressure by a batch process. No transfer of reaction products into transport or storage containers 20 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) PERCENT SULFIDE COMPOUND REMAINING TIME (HRS.) | REACTION WITH SONOCHEMISTRY } REACTION WITHOUT SONOCHEMISTRY Fic. 2. 2-Chloroethyl ethyl sulfide + nicotine. would need to occur until the products could be assessed for the presence of residual war- fare agent, thus providing this process with a level of safety not possible with incineration. We further propose that dedicated railroad tank-cars could be constructed that would serve both as “reaction vessels” and as a means to transport the reaction products to a toxic waste dump. This would minimize the number of times transfer of the chemicals would be required. (3) The proposed chemical demilitarization reagent nicotine will mix well with GB, VX and HD and thus improve the effectiveness of destroying these agents completely. All of these agents could be demilitarized in the same batch reactors (tank-cars), thus adding an additional economical benefit to this pro- cess. (4) A closed-loop batch reactor of the type that would be used for this process would not be useful for the destruction of most industrial toxic wastes and therefore would not serve to be converted into a general toxic waste dis- posal unit after the chemical warfare agents are demilitarized. (5) Chemical demilitarization of the agents would not produce TCDD, “dioxin.” (6) The reaction products are non-volatile salts or high-boiling liquids, substantially re- ducing any hazard in handling or transporta- tion. reaction with sonochemistry; © —reaction without sonochemistry. There are other non-chemical benefits that would be afforded by applying this demilita- rization method to the Chemical Stockpile Disposal Program: (1) The Department of Defense and its ex- ecutive agent for chemical stockpile disposal, the U.S. Army, would benefit. Their support of this project, or even support for a serious study of the method, would indicate a willing- ness to explore alternatives to the incineration program and thereby improve their credibility with the civilian populations surrounding the eight continental storage sites. This is no small matter since incidents at storage-disposal sites over the past few years have reduced their credibility and recent revelations about secret government tests with radioactive agents on civilians in the 1940s and 1950s will make it even more difficult for these agencies to con- vince the civilian population that concern for their health has a top priority. (2) The use of nicotine as the demilitarizing reagent in the destruction of the chemical warfare agents would provide an economic boon to the tobacco farmers not only of Ken- tucky, but also of several other tobacco-grow- ing states, some with depressed economies, as well as to the businesses and workers associ- ated with the recovery of nicotine from tobac- co. The work reported in this paper suggests that additional research needs to be accom- DECOMPOSITION OF NERVE AND MustTAarRD GasEs—Meisenheimer et al. al plished. Specifically, the warfare agents them- selves need to be used and the demilitariza- tion conditions optimized. In the process of optimizing the conditions, an effort should be made to verify the reaction mechanisms and to insure the absence of reversibility. In ad- dition, bioassays of the mixture of reaction products need to be made to determine gen- eral toxicity (LD.,) values. Reaction mixtures from mustard gas decomposition need to be assayed for residual vesicant activity and those from the nerve gases checked for residual an- tiacetylcholinesterase activity. ACKNOWLEDGMENTS The authors thank Mr. John Magera and Ace Glass Incorporated for their generosity in providing the sonochemical reactor used in this research. We thank the referee for his/her thoughtful suggestions. LITERATURE CITED 1. Ember, L. R. chemical weapons likely to be waylaid. Chem Eng News 70:19-20. 2. Bertazzi, P. A., A. C. Pesatori, D. Consonni, A. Ti- roni, M. T. Landi, and C. Zocchetti. 1993. Cancer inci- dence in a population accidentally exposed to 2,3,7,8-Te- trachlorodibenzo-para-dioxin. Epidemiology 4:398. 3. Hileman, B. 1993. Dioxin toxicity research; studies show cancer, reproductive risks. Chem Eng News 71:5-6. 4. Hileman, B. 1994. More regulation of dioxin sources sought. Chem Eng News 72:21-22. 1993. Alternate tech- nologies for the destruction of chemical agents and mu- nitions. pp. 3940. 6. Merck Index. 1989. 11:996. 7. National Research Council. 1994. Recommenda- tions for the disposal of chemical agents and munitions. p. 160. 8. National Institute for Occupational Safety and Health. 1976. Criteria for a recommended standard oc- cupational exposure to malathion. pp. 17-84. 9. Ley, S. V. and C. M. R. Low. 1989. Ultrasound in synthesis. Springer-Verlag, Berlin, New York. 1992. Army’s plans for destroying 5. National Research Council. Trans. Ky. Acad. Sci., 56(1—2), 1995, 22-27 Oocyte Staging in Paddlefish, Polyodon spathula WILLIAM L. SHELTON! Zoology Department, University of Oklahoma, Norman, Oklahoma 73019 AND STEVEN D. Mims Aquaculture Research Center, Kentucky State University, Frankfort, Kentucky 40601 ABSTRACT Oocyte size, germinal vesicle position, and pigment distribution of ova were examined as indicators of progress toward ovulation for hormonally injected paddlefish, Polyodon spathula. A boil and cut procedure to identify germinal vesicle position in oocytes was useful for estimating stage of oocyte maturation. Pigment pattern was a functional alternative for identifying the germinal vesicle position, because it could be substi- tuted for the boil and cut procedure. Ova size was of no practical significance in relation to estimating potential spawning success or ovulation time. Latent time to ovulation for females injected with Luteinizing- Hormone-Releasing Hormone analogue was shorter than that of females injected with paddlefish pituitary extract. Information on the rate of germinal vesicle migration relative to water temperature would facilitate prediction of ovulation time following hormonal injection. INTRODUCTION Artificial propagation of paddlefish is an im- portant tool for resource management. It may be used to enhance natural populations (1) or to produce stock for aquaculture (2). Natural stocks have been impacted by environmental perturbations and by recent exploitation for the caviar market (3, 4). Some governmental agencies are assessing the status of various natural populations, and are developing plans to revitalize the affected populations. In most warmwater species, ova viability de- teriorates relatively quickly after ovulation. Consequently, the quality of ova produced by induced spawning can be improved by the ability to accurately predict time of ovulation. Convenience to hatchery personnel can also be enhanced. Techniques used to examine in- tra-ovarian eggs are related to species specific characteristics. During final maturation of oo- cytes, cytological reorganization is usually as- sociated with visible changes. The usefulness of these changes as predictors of ovulation de- pends on the ability to relate identifiable stan- zas with time-related sequences (5). An im- portant event associated with these changes is migration of the germinal vesicle (GVM) to- ' Correspondent author. 22 ward the periphery of the oocyte to complete the first meiotic division (6, 7, 8). Coalescence of oil globules and related re- distribution of cellular components results in spontaneous ova clearing in many marine fish- es. For example, this has been used as an im- portant tool to predict ovulation in striped bass, Morone saxatilis (9). However, cytologi- cal redistribution and germinal vesicle (GV) position are not easily viewed in ova of most freshwater species because their oocytes are opaque. The yolk of some fish oocytes can be cleared with one of several solutions so that the germinal vesicle can be seen (10). How- ever, this method is not satisfactory for oocytes of fish species that have pigmented eggs. Lutes et al. (11) located GV positions in the eggs of white sturgeon, Acipenser transmon- tanus by heat-hardening the oocytes and cut- ting them along the animal-vegetal axis. The objectives of this study were to test the appli- cability of the technique used to identify GV position in sturgeon oocytes, in paddlefish oo- cytes, and to examine other functional alter- natives. MATERIALS AND METHODS Initially, a seasonal series of oocytes was sampled from wild adult paddlefish in Grand OocyTEs IN PADDLEFISH—Shelton and Mims 23 Lake, Oklahoma (12). Oocytes and pituitaries were collected from fish being processed for caviar by licensed commercial fishermen. Pi- tuitaries were frozen for later use in spawning, while ova were examined fresh. Ten oocytes from each mature female were measured along the vertical axis, the pigment distributions were noted, and then the oocytes were boiled for 3—5 minutes to harden them. Similar to the PSE, used for sturgeon by Lutes et al. (11), the boiled oocytes were bisected through the animal-vegetal pole axis, and the position of the GV was recorded. Some ova were preserved in formalin (5—10%) for several weeks to see if they would harden sufficiently for cutting. Sever ral clearing solu- tions were also used eh fresh oocytes to test their effectiveness to provide a view of the GV. In a few tests, commercial bleach was used as a follow-on treatment to a clearing solution. The solutions included an alcohol-xylene treat- ment (13), Stockard’s solution (14), and a serra solution (15). Stoeckel and Neves (10) found these solutions to be effective in clearing oo- cytes of 9 teleosts. The serra solution has been widely used in Europe for clearing oocytes of common carp, Cyprinus carpio (16). Artificial propagation trials were conducted at the Aquaculture Research Center, Ken- tucky State University. Broodstock in spawn- ing condition were netted and transported to the hatchery facility, where they were hor- monally injected following the procedures of Graham et al. (17) and Semmens (18). Fresh- frozen paddlefish pituitaries or super-active analog of Luteinizing Hormone—Rele easing Enon. LH-RHa (Sigma) were used as in- ducing agents. A priming dose (1/10 of the to- tal) and a resolving dose (9/10) were admin- istered 12-24 hr apart. Intra-ovarian eggs were sampled through a small ventral incision as described by Doroshov et al. (19) at the time of priming and in some instances at the resolving injection. Pigmentation patterns and GV positions of these oocytes were examined. RESULTS AND DISCUSSION Oocyte Treatments Ova were examined from 93 mature (>15 kg) females monthly from November through April (Table 1). Oocyte size spanned nearly the same range in November as in April just TABLE 1. ovarian samples during the prespawning period. Paddlefish oocyte size collected from fresh Mean height Month (mm) Range Number of fish (n = 10 ova) November 2.46 DEVS. 52) 20 December 2.49 DB) /F/ 1S January Dia) 2.44-2,.89 14 February 2.63 2. 48-294 20 March 2.70 2.31-2.92 15 Spawning Tal 2.56—2.79 6 prior to spawning. There was a slight increase in mean size from November through April, but this change would be of no practical value in staging oocytes for induced ovulation. Ova size during the prespawning period was simi- lar to that reported for paddlefish in Louisiana (20). Heat-hardening of paddlefish oocytes and subsequent bisection resulted in 2 egg halves that were satisfactory for locating chen GV. The GV could be clearly seen as a fine-textured, round object within the more granular yolk. None of the formalin-fixed oocytes were hard- ened enough to be cut without distorting the Further, none of the solutions cleared paddlefish ova so that the GV could be seen because the dark pigment screened any change in yolk opacity. Treatment with bleach Sontewiht ‘reduced the pigment intensity, but also distorted the oocytes so that they could not be used for staging. egg, Oocyte Staging In the period just preceding natural spawn- ing, the GV was in a central position (Fig. 1, right couplet of Stage I) os pigment was un- equally distabuted | (Fig. left couplet of Stage I). Pigment appears es be more concen- fated near nthe oocyte periphery rather than being dispersed i in the yolk. Yolk in the animal hemisphere is more uniform in particle size, than that of the vegetal hemisphere. The an- imal hemisphere also has a much greater con- centration of pigment than the vegetal hemi- sphere. The latter is whitish-grey, while the former is charcoal colored. Transition of pig- ment distribution at the equatorial circumfer- ence is distinct, and often appears as a shad- owy ring at the equator. Position of the cen- trally pened GVs usually corresponds to the zone of transition between the vegetal and an- 24 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) Hrewwle iD) Paddlefish oocyte stages (I-VI); the left illustration of each pair is representative of the surface pigmentation and the right illustration of each pair is of a bisected oocyte showing yolk distribution and germinal vesicle (GV) position. Stage I typifies an oocyte with a centrally located GV prior to resumption of meiosis-I; Stage I illustrates the slightly displaced GV at the initiation of germinal vesicle migration (GVM) and pigment redistribution after the re- sumption of events leading to meiosis-I; Stage III shows the GV in a position in the upper region of the oocyte near the animal pole; Stage IV depicts the clearing of the animal pole region in relation to the GV position; Stage V is characterized by further pigment change and the disappearance the GV at germinal vesicle breakdown (GVBD); Stage VI shows the appearance of an ovulated egg. OC imal hemisphere pigmentation. This relation- ship is maintained as the GV migrates toward the animal pole. The GV was in the central position (Stage I) throughout the fall and early spring. Migra- tion toward the animal pole occurred rapidly, and just prior to ovulation. As the GV migrates (GVM), the pigment discontinuity also moves poleward (Fig. 1, Stage II). Pigment near the transition is often more concentrated, giving the appearance of a ring. The position of the pigment margin does not continue to move as OocyTES IN PADDLEFISH—Shelton and Mims TABLE 2. Oocyte stage at injections* Ovulating Prime Interval (hr) Resolve Agent** I 24, 24*** II, Ul P II DAW ALS III, IV P Il 24 — le IV 22, = P Ill 24 = IE; Il 12 IV L III-IV ily V IL I1I-IV 22 i Ib IV 22 — IG, * See Figure |. ** P = fresh-frozen paddlefish pituitaries; L = LH-RHa *** Second resolving injection. the GV shifts further to approximately %—% the distance to the pole (Fig. 1, Stage HI) and there is a more uniform ark pigmentation, rather than the ring-like pattern seen earlier. However, Stage-II and Stage-III oocytes are difficult to differentiate wathout direct refer- ence to the GV position. As GVM continues, a clear area appears at the animal pole (Fig. 1, Stage IV). Consequently, at Stage IV, a rel- atively broad band of pigment occupies the middle two-thirds of the animal hemisphere and the GV is now located at the polar edge of the pigment band. Appearance of the clear area at the animal pole is indicative of Ger- minal Vesicle Breakdown (GVB), and subse- quently completion of the first meiotic division and ovulation. The clear area of the oocyte contracts slightly prior to ovulation, and the GV is no longer visible in gross cross-section (Fig. 1, Stage V). The ovulated egg (Fig. 1, Stage VI) has a distinct elevated pr obuberance at the animal pole/micropylar area, and the pigment is slightly more diffuse than that of earlier stages. Application to Propagation Direct observation of oocyte GV position, or the changing pattern of pigmentation can be used to stage oocytes of paddlefish. Position of the GV was used as an indication of matura- tional activity, once GVM had been initiated. However, because the GV is centrally located for most of the prespawning period these characteristics are useful as indicators of ap- proximate time to ovulation only after some displacement has occurred. Oocytes from females collected early in the spring-time spawning cycle were typically in SS) Ot Paddlefish oocyte stage, inducing agent, and temperature related to latent period for ovulation. Latent period (hr) Temp Ovulated/ (C) injected Resolve Total 13-14 0/2 both died 13-14 2/2 18-19 66 13—14 1/] 25 49 17-18 1/] 18 4() 13-14 V/3 15 39 17-18 6/6 24-96 36-38 15-16 8/9 24-28 3640, 18-19 5/5 19 3] 17-18 2/2 13:5 645) Stage I or II. Some females that were collect- ed when spring-time ambient temperatures were approaching the spawning range, and that had oocytes with a centrally foeated GV (Stage I), were not ovulated even after mul- tiple injections (Table 2). Oocyte maturation (GVM) was stimulated by two resolvi ‘ing injec- tions but these fish died before ovulation oc- curred. However, when GVM was proceeding (Stage II), responsiveness to induction was more predictable. Twenty four of 28 females with oocytes at Stage II or IV were induced to ovulate within 31 to 49 hr after the priming injection. Two factors, temperature and induc- ing agent, appeared to influence the timing of ovulation. Because temperature affects maturational rate of oocytes in fishes, incorporation of a temperature component could improve pre- dictive power of time to ovulation of hormon- ally injected paddlefish. Only general infer- ences can be made from the present data. Higher temperatures within the normal spawning range did result in shorter times to ovulation (Table ‘ 2). Temperature influence on developmental rate for the Russian sturgeon, Acipenser guldenstadti, is described by @ine berg and Dettlaff (21). Staging under different temperature regimes may provide more in- sight into establishing a GVM rate and im- prove the usefulness; however, the inducing agent must also be considered. In our study, the latent period for females injected with LH-RHa was generally shorter than that of females injected oa pituitary eXx- tract. Further, the response to LH-RHa stim- ulation appeared to be less temperature sen- 26 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-2) sitive than that of pituitary induction. Sem- mens (18) also reported shorter latent periods for paddlefish injected with LH-RHa com- pared to pituitary stimulation. The reciprocal response would be expected based on the pathways and modes of action of the 2 agents. Rottmann and Shireman (22) reported that la- tent time to ovulation of Chinese carps was longer for LH-RHa than for pituitary induc- tion. CONCLUSIONS Environmental factors influence the prog- ress of gamete maturation. Intraovarian sam- ples can provide a window to the process and indicate the status of the maturational trajec- tory. Whether a female is responding to an in- ducing agent, or will need additional stimulus to ovulate are pertinent questions during ar- tificial propagation. The position of the GV, or in the case of paddlefish, the ova pigment pat- tern, can be used to provide more information during induction, or can be used as tools to examine more detailed relationships. In the latter context, a GVM-rate/temperature rela- tionship would be useful in the artificial prop- agation of paddlefish. With this information, other factors, such as a more thorough com- parison of the efficacy of inducing agents could be examined. Applicability of staging techniques may provide useful tools in facili- tating artificial propagation of related fishes threatened with extirpation such as the Chi- nese Paddlefish, Psephurus gladius. ACKNOWLEDGMENTS Field support from biologists of the Okla- homa Department of Wildlife Conservation and from commercial fishermen in both Okla- homa and Kentucky is gratefully acknowl- edged. Financial support was provided by two grants, USDA 83-CRSR-2-2228 to the Uni- versity of Oklahoma and USDA/CRS KYX-80- 85-01A to Kentucky State University. LITERATURE CITED 1. Graham, L. K. 1986. Establishing and maintaining paddlefish populations by stocking. Pp. 95-104. In J. G. Dillard, L. K. Graham, and T. R. Russell (eds.) The pad- dlefish: status, management and propagation. Amer. Fish. Soc., N. Cent. Div., Spec. Publ. 7. 2. Semmens, K. J. and W. L. Shelton. 1986. Oppor- tunities in paddlefish aquaculture. Pp. 106-113. In J. G. Dillard, L. K. Graham, and T. R. Russell (eds.) The pad- dlefish: status, management and propagation. Amer. Fish. Soc., N. Cent. Div., Spec. Publ. 7. 3. Pasch, R. W. and C. M. Alexander. 1986. Effects of commercial fishing on paddlefish populations. Pp. 46-53. In J. G. Dillard, L. K. Graham, and T. R. Russell (eds.) The paddlefish: status, management and propagation. Amer. Fish. Soc., N. Cent. Div., Spec. Publ. 7. 4. Sparrow, R. D. 1986. Threats to paddlefish habitat. Pp. 36-45. In J. G. Dillard, L. K. Graham, and T. R. Russell (eds.) The paddlefish: status, management and propagation. Amer. Fish. Soc., N. Cent. Division, Spec. Publ. 7. 5. Shelton, W. L. 1979. Management of finfish repro- duction for aquaculture. CRC Rev. Aquat. Sci. 1:497—535. 6. Nagahama, Y. 1983. The functional morphology of teleost gonads. Pp. 223-275. In W. S. Hoar, D. J. Randall, and E. M. Donaldson (eds.) Fish physiology, Vol. 9. Ac- ademic Press, New York. 7. Dettlaff, T. A. and S. G. Vassetzky (eds.) 1988. Oo- cyte growth and maturation. Consultants Bureau, New York. 8. Saat, T. 1993. The morphology and chronology of oocyte final maturation and fertilization in fish. Pp. 71— 85. In B. T. Walther and H. J. Fyhn (eds.) Physiological and biochemical aspects of fish development. University of Bergen, Norway. 9. Stevens, R. E. 1965. A final report on the use of hormones to ovulate striped bass, Roccus saxatilis (Wal- baum). Proc. Southeast. Assoc. Game and Fish Comm. 18 (1964):525-538. 10. Stoeckel, J. N. and R. J. Neves. 1992. Comparison of methods for viewing the germinal vesicle in fish oo- cytes. Progr. Fish-Cult. 54:115-115. 11. Lutes, P. B., S. I. Doroshov, F. Chapman, J. Harrah, R. Fitzgerald, and Martin Fitzpatrick. 1987. Morpho- physiological predictors of ovulatory success in white stur- geon, Acipenser transmontanus Richardson. Aquaculture 66:43-52. 12. Combs, D. L. 1982. Angler exploitation of paddle- fish in the Neosho River, Oklahoma. N. Amer. J. Fish. Manag. 4:334—-342. 13. Chen, F. Y., M. Chow, and B. K. Sim. 1969. In- duced spawning of three major Chinese carps in Malacca, Malaysia. Malay. Agric. J. 47:211-238. 14. DeMontalembert, G., B. Jalabert, and C. Bry. 1978. Precocious induction of maturation and ovulation in northern pike (Esox lucius). An. Biol. Anim. Bioch. Bio- phy. 18:969-975. Ws), Levavi-Zermonsky, B. Z. Yaron. 1986. Changes in gonadotropin and ovarian steroids associated and with oocytes maturation during spawning induction in the carp. Gen. Comp. Endocrin. 62:89-98. 16. Shelton, W. L. In press. Sex control in carps. Sym- posium on carp genetics, 3-7 September 1990. Szarvas, Hungary. 17. Graham, L. K., E. J. Hamilton, T. R. Russell, and C. E. Hicks. 1986. The culture of paddlefish—a review OocyTEs IN PADDLEFISH—Shelton and Mims 7 of methods. Pp. 78-94. In J. G. Dillard, L. K. Graham, and T. R. Russell (eds.) The paddlefish: status, manage- ment and propagation. Amer. Fish. Soc., N. Cent. Div., Spec. Publ. 7. 18. Semmens, K. J. 1986. Evaluation of paddlefish hy- pophysis, carp hypophysis and LHRH analogue to induce ovulation in paddlefish, Polyodon spathula. Ph.D. disser- tation. Auburn University, Alabama. 19. Doroshov, S. I., W. H. Clark, P. B. Lutes, R. L. Swallow, K. E. Beer, A. B. McGuire, and M. D. Cochran. 1983. Artificial propagation of the white sturgeon, Aci- penser transmontanus Richardson. Aquaculture 32:93— 104. 20. Reed, B., W. E. Kelso, and D. A. Rutherford. 1992. Growth, fecundity, and mortality of paddlefish in Louisi- ana. Trans. Amer. Fish. Soc. 121:378-384. 21. Ginsberg, A. S. and T. A. Dettlaff. 1991. The Rus- sian sturgeon Acipenser guldenstadti. Part I. gametes and early development up to hatching. Pp. 15-65. In T. A. Dettlaff and S. G. Vassetzky (eds.) Animal species for de- velopmental studies, Vol. 2. Vertebrates. Consultants Bu- reau, New York. 22. Rottmann, R. W. and J. V. Shireman. 1985. The use of synthetic LH-RH analogue to spawn Chinese carp. Aquacult. Fish. Manag. 1:19-24. Trans. Ky. Acad. Sci., 56(1—2), 1995, 28-40 Egg and Larval Development of the Striped Fantail Darter, Etheostoma flabellare lineolatum (Agassiz), and Duskytail Darter, E. Percnurum Jenkins, with Comments on the Etheostoma flabellare Species Group THOMAS P. SIMON! University of Wisconsin—La Crosse, Department of Biological Sciences, La Crosse, Wisconsin 54601 AND STEVEN R. LAYMAN University of Alabama, Department of Biological Sciences, Box 870344, Tuscaloosa, Alabama 35487-0344 ABSTRACT The Etheostoma flabellare species group comprises 6 recognized forms, including E. f: lineolatum and E. percnurum. The wide ranging E. f. lineolatum and the relict E. percnurum both exhibit large spherical yolk sacs, with a mid-ventral vitelline vein plexus; 34-36 total myomeres; well developed jaws at hatching; and head not deflected over the yolk sac. Eggs of E. f. lineolatum range between 2.0-2.8 mm in diameter. Hatching in E. f. lineolatum occurs between 4.5-5.9 mm TL and first ray formation occurs at 5.0 mm in the pectoral fins. Etheostoma percnurum has the largest eggs of the group with diameters between 2.6-3.3 mm. Larvae hatch between 4.5-5.4 mm TL and possess six incipient rays in the pectoral fin. Fin-ray formation occurs at later length intervals than E. f. lineolatum, except for the spinous dorsal fin. Orbit shape and initiation of squamation for E. percnurum is similar to E. kennicotti. The fantail darter, Etheostoma flabellare, is the most widespread member of the subgenus Catonotus, ranging from the Great Lakes ba- sin, south through the upper and middle Mis- sissippi basin, and east onto the central Atlan- tic slope (1). This highly variable taxon, con- sists of 3 to 5 subspecies. However, E. f. li- neolatum, was not recognized by McGeehan (2) because of the erratic geographical distri- bution of striped and non-striped forms. Eth- eostoma flabellare inhabits gravelly or rocky riffles (occasionally slow runs or pools) in headwater creeks to moderate-sized_ rivers, and is potentially sympatric with at least 11 other species of Catonotus (1, 3, 4). In con- trast, the duskytail darter, Etheostoma perc- nurum, is a relict species restricted to only 4 large streams of the upper Tennessee and middle Cumberland River drainages, where it inhabits rocky pools (4, 5). It is accorded threatened status by the state of Tennessee (6), and endangered in Virginia (7, 8). It oc- ' Present address: Large Rivers Research Station, 119 Diana Road, Box 96, Ogden Dunes, Indiana 46368. 28 curs sympatrically with only 1 species of Ca- tonotus, E. flabellare. The fantail and duskytail darters represent distributional extremes with- in Catonotus, but are closely related, compris- ing, with E. kennicotti, the E. flabellare species group (3, 9). There is a growing body of comparative in- formation for distinguishing larvae and juve- niles of different species of Catonotus (10, 11). Embryonic and larval development of E. fla- bellare flabellare, the subspecies occupying the Great Lakes and upper Ohio River basins (2), has been characterized by Lake (12), Coo- per (13), Auer (14), and Paine (15); however, descriptions are lacking for other subspecies. Also, given the high degree of sympatry be- tween E. flabellare and other species of Ca- tonotus, comparative data are needed for sep- arating their larvae. Layman (5) described lar- val stages of the duskytail darter but did not report his observations of embryonic devel- opment or provide meristic and morphometric characteristics of larvae. Such information may become critical in implementing future recoy- ery plans for this jeopardized species. Larval Ecc AND LARVAL DEVELOPMENT IN Darters—Simon and Layman characters may also prove to be useful in re- solving phylogenetic relationships within the E. flabellare species group. This paper describes larval development of Etheostoma flabellare lineolatum, the striped fantail darter, a widespread taxon found in the lower Ohio River basin and tributaries of the Mississippi River, and embryonic and larval development of E. perenurum, the duskytail darter. Meristic, morphometric, and pigmen- tation features are used to separate these taxa from E. f. flabellare, E. kennicotti, and other sympatric species of Catonotus. MATERIALS AND METHODS Laboratory cultured and wild collected specimens were studied for differences in morphology, meristics, pigmentation, and se- quential development relative to size. A series of meristic and morphometric features was measured from 128 eggs and 237 larvae and early juvenile striped fantail darters, and 20 eggs and 24 larvae and early juvenile duskytail darters. A total of 21 morphometric and nine me- ristic characters was measured for each spec- imen following methods outlined in Simon (10). Eggs and embryos were preserved in 5% formalin, while larvae were preserved in 10% formalin after removal from nests. The middle of the spawning episode (i.e., fertilization of ova) was used as time zero in estimating ages of embryos. All measurements were made to the nearest 0.1 mm using a dissecting micro- scope with an ocular micrometer. Measure- ments in the text are expressed asa proportion of total length (TL) unless otherwise noted. Illustrations were delineated following Sumida et al. (16). Embryonic and larval descriptions of the duskytail darter were based on adults collected and spawned by Layman (5) from the Ten- nessee River drainage, Little River, Blount County, Tennessee. Little River contains 1 of 3 extant populations for the species in the up- per Tennessee River drainage. Eggs were ob- tained from nests in Little River and from spawnings of aquarium- -held adults. All eggs were incubated in aquaria at 18—27°C. Larval descriptions of striped fantail darter were based on adults collected and reared by Simon (17) from the Mississippi River drainage, Spring Coulee Creek, Vernon County and 29 a Coon Creek, La Crosse County, Wisconsin; Root River, Fillmore County, Minnesota: and upper lowa_ River, Houston County, Lowa. Eggs were obtained from spawnings of aquar- ium-held adults and incubated at 2 tempera- tures (20° and 23°C) in the laboratory. Larvae of both species were fed live Artemia nauplii. SPECIES ACCOUNTS Striped fantail darter, Etheostoma flabellare lineolatum (Agassiz) Eggs.—Three size classes of ova were ob- served in the dissected ov ary of E. flabellare lineolatum. The smallest ova ranged 1.0-1.5 mm (n = 13, = 1.3 mm), and were spheri- cal, opaque, and pale yellow. The intermediate sized ova were ovoid, opaque, and pale } yellow and ranged 1.5-2.0 mm (n = 15, * = 1. 6 mm). Mature eggs collected from Spring Coulee Creek, Wisconsin, the Rock River, Illinois, and the upper Iowa River, Iowa, were all equiva- lent in diameter and ranged from 2.0-2.8 mm (n = 100, = 2.4 mm). Mature eggs are spherical, demersal, and adhesive with trans- lucent yellow yolk. Generally, mature eggs have a single oil globule, a narrow perivitelline space, an “unsculptured chorion, and are un- pigmented. Based on presence of various size classes of ova and aquarium observations, the striped fantail darter is considered a multiple spawner depositing more than a single clutch of eggs Eggs are attached to the undersides of slab rocks in the mar gins and slower portions of riffles and raceways (2, 12, 18, 19, 20). Embryonic Development. —Embryonic de- velopment for the fantail darter has been pre- viously described by Lake (12), Cooper @is)t and Paine (15). Eggs incubated at 20°C hatched after 240 + “T8 hr (9.25-10.75 days); at 23°C, hatching occurred in 144 + 10 hr (5.50-6.50 days). Lake (12) reported eggs of Etheostoma f. ‘flabellare to hatch in 30-35 days at 17°-20°C; after 21 days at 21°-22°C; and in 14-16 days at 26°C. Larvae Morphology.—The size of initial formation for selected structures is summarized for larval and early juvenile E. f. lineolatum with mor- phometric features shown in Table 1. At 4.5— 5.9 mm, newly hatched, well dev eloped pec- 30 sample size). Characters Morphometry of Etheostoma flabellare lineolatum larvae and early juveniles grouped by selected intervals of total length (N expressed as percent total length or head length with a single standard deviation. TABLE lL. Depth (% TL) Length (% TL) Caudal Peduncle Anus 10.4 + 1.4 Shoulder 32.2 + 4.9 Head 196 + 1.4 Head 18.4 + Preanal Snout Eye Standard 89.7 + N Total length 6.5 + 0.8 al ol fo] a [oy 61 147 4.5-6.9 + | 16.1 + 3.6 AN SIlL@ ae Iky A 0.4 NN 85.4 + 7.0-12.8 13.0-15.9 16.0-18.5 19.1- GS) ae -O,7/ 10.0 14.0 + 0.9 14.6 + 1.1 t ISKoy as IU 16.7 + 1.8 06) ae Ils +| be al (oe) 16 0.8 +l 16.6 + 04 Wi 4 4 5 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-—2) 9.4 + 0.9 9.5 14.6 + 0.7 13.8 + 1.0 0.8 + 0.3 13.4 + 0.8 21.6 Il 4 16.4 + 1.2 20.6 + 1.9 50.3 + 0.9 86.1 + 3.5 toral fins without incipient fin rays; yolk sac extremely large, spherical (ca. 42% TL); yolk amber, with a single anterior oil globule; vi- telline vein forming a plexus on mid-ventral yolk sac; head not deflected over the yolk sae; jaws developed; eye diameter oval. Notochord flexion occurs 4.9-6.0 mm; first rays form in pectoral fin (5.0-5.6 mm); anal fin rays (5.3— 5.9 mm); first caudal fin ray form (5.3-5.9 mm); soft dorsal rays form (5.4-6.2 mm); and spinous dorsal rays form (5.9-6.2 mm). Caudal fin round (5.8—6.2 mm); incipient dorsal and anal fin margins partially differentiate (7.1—8.0 mm); spinous dorsal fin origin situated over preanal myomere 4, soft dorsal origin over preanal myomere 15-16 (7.8 mm); predorsal length 36.3% SL (range: 33-39.7% SL); anal fin margin completely differentiate (7.9 mm); pelvic fin buds form anterior to dorsal fin or- igin prior to complete absorption of yolk sac (7.9-8.5 mm); yolk absorbed (7.8-8.0 mm). Entire finfold absorbed by 8.2 mm; first pelvic fin rays form (8.8—-9.2 mm); no swim bladder forms, remains rudimentary (trace presence); gut straight; squamation initiated at 10.0 mm. Preoperculomandibular canals form (10.8— 11.8); supraorbital, infraorbital, and_ lateral head canals form (11.0-12.6 mm); supraorbital completely form (11.7 mm); preoperculoman- dibular pores 10, completely form (12.6—13.8 mm); infraorbital canal form with 8 pores (10.8 mm), completely form with retrogres- sion to interrupted condition of 4 pores ante- riorly and 2 pores posteriorly (14.4-14.6 mm). Lateral line forms (14.0-14.2 mm); squama- tion complete (14.7 mm); scales absent on the nape, cheek, opercle, breast, and prepectoral areas. Meristics—Myomere number in E. f. lineo- latum is constant posthatching, preanal myo- meres 15, postanal myomeres 19-21 (¢ = 19.5; n = 155), with 34-36 total myomeres. Total vertebrae number 33-34 (< = 33.8, n = 5), including one urostylar element (from cleared and stained specimens from Spring Coulee Creek and the Root River). Scales in the lateral series ranged from 42-57 (* = 47.7; mode = 48: n = 13). Paired and median fin rays and length at appearance are summarized in Table 2. Pigmentation.—Newly hatched larvae with scattered melanophores on a large yolk sac with greatest concentration laterally. Stellate EGG AND LARVAL DEVELOPMENT IN DarTERS—Simon and Layman 3] TABLE 2. Selected meristic values and size (mm total length) at the apparent onset of development for Etheostoma flabellare lineolatum and E. percnurum. Mean values are underscored. The number of secondary rays of the median fins are in lowercase Roman numerals. Attribute/Event E. f. lineolatum E. percnurum Dorsal fin spines/rays VITI-IX/12-13-14 VI-VII-VIMI/10-12-13 First rays formed 5.9/5.7) 5.8/5.8 Adult complement formed REOMEZ 6.1/7.4 Pectoral fin rays Pais} 121314: First rays formed 5.0-5.6 5.8 Adult complement formed 7.0 6:1=6:2 Anal fin spines/rays I1/9-10-11 II/7-8-9 First rays formed CaS) 5.8 Adult complement formed WV=12 7A Pelvic fin spines/rays I/5 V/5 Bud formed 7.9-8.5 >8.1 First rays formed 8.8-9.2 93 Adult complement formed 8.8-9.2 9.3 Caudal fin rays ix-xiv, 8 + 7, viii—xiv xi-_xvi, 8 + 8-9. xi-xv First rays formed By) 5.8 Adult complement formed COHWE 15) Lateral series—scales 42-48-57 3840-45-48 Myomeres/vertebrae 34-36/33-—34 34-36/: 33-34-35 Preanal myomeres Tels i Postanal myomeres 19-21 19-21 melanophores encircle optic lobe. Several me- lanophores laterally, rising obliquely near nape; lateral pigmentation with a midlateral stripe from the yolk sac posterior to base of caudal peduncle. Ventral melanophores pres- ent on gut and from posterior anus to approx- imately , postanal myomere 9. Dorsal melano- phores in 2 blotches located just anterior the anus, and initiating near posterior of ventral postanal melanophores. Majority of preanal myomeres without pigmentation (4.5-5.9 mm; Fig. 1). Postorbital bar formed, with additional horizontal pigment on operculum. Yolk sac with stellate melanophores on distal half. Dor- sally, pigmentation on nape and at the base of soft dorsal. Laterally, melanophores outlining preanal myosepta posteriad of yolk sac, ex- tending to middle of soft dorsal: a midlateral stripe formed from single melanophores. at apex of preanal myomeres posterior of yolk sac. Ventrally, stellate melanophores present at almost every postanal myoseptum with several extending dorsally to midlateral; melano- phores present at base of caudal fin (6.0-7.5 mm; Fig. 2A). Horizontal preorbital and post- orbital bars with additional pigment present on operculum, and dorsally on nape; melano- phores outline lateral myosepta of all myo- meres just posteriad of soft dorsal. Melano- phores extend onto rays of caudal, anal, and soft dorsal. Ventral pigmentation concentrated at midventral gut, and beneath operculum and branchiostegal rays (7.8-9.2 mm: Fig. 2B). Chevron shaped clusters present dorsal-ante- rioriad to orbit, and on the optic lobe. Dorsally 8 rectangular blotches extend from nape to base of caudal peduncle. Oval blotches be- come continuous anteriorly along the midlat- eral with scattered gut melanophores. Ventral pigmentation limited to 5 areas of concentra- tion from just after anus to base of caudal pe- duncle. Spinous dorsal, pectoral, pelvic, and anal fins devoid of pigment (9.5-10.9 mm; Fig. 3A). An oblique bar extends towards nape lat- erally, posterior the orbit; cerebrum and optic lobe with clustered melanophores. Dorsally, 9-10 rectangular blotches with obliquely scat- tered melanophores connecting 12—13 midlat- eral blotches. Lateral epaxial scales outlined with scattered melanophores; pectoral girdle with a blotch near cleithrum. Lepidiotrichia of spinous and soft dorsal, anal, and base of cau- dal fins with melanophores. Mandible, maxilla, and interopercle with scattered melanophores; pectoral and ee fins devoid of pigment (11.0-13.9 mm). Juvenile pigmentation, cra- oo Trans. KENTUCKY ACADEMY OF SCIENCE 56(1—2) mm an c. Fic. 1. Etheostoma flabellare lineolatum, striped fantail darter, newly hatched yolk sac larva, 4.8 mm TL, Coon Creek, Wisconsin. a. dorsal, b. lateral, c. ventral views. EGG AND LARVAL DEVELOPMENT IN DartERs—Simon and Layman 33 BTN og he ie Fre. 2. lateral view, b. 8.2 mm TL larva, lateral view. nium with concentrated melanophores over optic lobe and cerebrum; distinct preorbital and postorbital bars formed, no suborbital tear drop; a chevron shaped cluster of melano- phores parallel to postorbital bar, scattered melanophores on cheek. Lateral pigmentation including 11-13 rectangular blotches connect- ing to § dorsal bands; the final lateral blotch may be divided to form 2 spots near midlateral caudal peduncle base. Horizontal lines of me- lanophores extend from the head to caudal pe- duncle, formed from individual melanophores on outer margins of scales; distinct humeral spot formed near posterior of opercular spine. Spinous dorsal, pectoral, and anal fins with scattered pigmentation on rays; 4—5 horizontal stripes on soft dorsal distributed on fin rays; caudal fin with 6-8 vertical stripes formed on Etheostoma flabellare lineolatum, striped fantail darter, Spring Coulee Creek, Wisconsin. a. 7.1 mm TL larva, interstitial membranes; pelvic fins without pig- mentation (Fig. 3B). Duskytail darter, Etheostoma percnurum Eggs.—Nests of the duskytail darter in Lit- tle Fiver consisted of single- layer clusters of 23 to 200 eggs (n = 22: = 79: SD = 46) attached to the undersides of slab-shaped stones (4, 21). Fertilized eggs were translucent and spherical, and averaged 2.8 mm diameter from wild nests (range: 2.63.3 mm: n = 35: SD = 0.2 mm) and 2.9 mm from aquarium nests (range: 2.6-3.3 mm; n = 20; SD = 0.1 mm: 4). The chorion was clear, adhesive, and flattened at the point of attachment to the nest stone. The yolk was translucent, and a large translucent amber oil globule, surrounded by 34 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) ° “» @ Cae 9, ° Podesta a On 08 ose... apes. bbe 2 ce ot - CD a4 @ Win -C . ° - = Fic. 3. lateral view. b. 14.4 mm TL juvenile lateral view. several much smaller ones, imparted an amber color to the egg. Embryonic Development.—The morula was round and measured 0.8—0.9 mm in diameter; yolk was 2.2—2.3 mm in diameter, with a single large oil globule 0.6—-0.7 mm in diameter; the perivitelline space ranged 0.1-0.4 mm (5 hr embryos). The blastoderm of the early embryo covered 67% of the yolk, the embryonic axis was forming, and the germ ring was visible (26 hr embryo). In the tail-bud stage, the optic vesicles were forming, and 9 somites were vis- ible (ca. 28-47 hr embryos). In early tail-free embryos, dorsal and ventral finfolds were de- Etheostoma flabellare lineolatum, striped fantail darter, Root River, Minnesota. a. 10.3 mm TL early juvenile, veloping; auditory vesicles were visible; lenses were forming in the unpigmented eyes; and about 30 somites were discernible; the heart was Clearly beating, and the embryo occasion- ally twitched. Melanophores were widely scat- tered over the yolk membrane and were most concentrated on the vent and body-yolk junc- ture (50 hr embryos). By the late tail-free stage, the head was highly elevated; the eyes were grayish-black; and the pectoral fin buds were present. Melanophores were more con- centrated over the yolk membrane and had developed ventrally along the body to the tail (72-96 hr embryos). In late embryos, the eyes sample size). Characters expressed Morphometry of Etheostoma percnurum larvae and early juveniles grouped by selected intervals of total length (N as percent total length or head length with a single standard deviation. TABLE 3. Depth (% TL) Length (% TL) Caudal peduncle Shoulder Anus Head Head Eye“ Snout" Preanal 56.8 + 4.6 BiO}ro} ae Ih, Standard 96.3 + 0.4 SOrone= N Total length EGG AND LARVAL DEVELOPMENT IN DARTERS—Simon and Layman 35 6 16 4.5-6.2 7 0.5 0) 35 t 15.0 ss lo | A-11.8 12.3-15.5 (6}(6}, as) (0), 1 2 5.1 4 30.8 Niffso) a= (0). were pigmented black; the newly formed mouth opened and closed; ope srcles_ moved: and the well-developed pectoral fin buds flut- tered. A highly branched vitelline plexus cov- ered the anterio-ventral portion of the yolk sac, very similar to that described by Paine (15) for embryos of E. f. flabellare. Melano- phores were present on the yolk sac, dorsally and ventrally along the bases of the finfolds, mid-laterally toward the tail, and on top of the head. Late embryos wiggled frequently and vigorously, and the chorion was soft and deli- cate (>125 hr embryos). Eggs hatched in 264— 336 hr (11-14 days) at 18°-27°C (5). Larvae Morphology.—The size of initial formation for selected structures is summarized for larval and early juvenile E. percnurum with morpho- metric features shown in Table 3. At 4.5—5.4 mm TL, newly hatched, pectoral buds were present with 6 incipient rays; first pectoral rays form (5.8 mm); yolk sac robust, spherical ca. 47.9% TL, yolk translucent, single oil globule 0.7 mm diameter; vitelline vein form a plexus on midventral yolk sac; head not deflected over yolk sac; jaws developed; eye diameter spherical. Median fin rays in the spinous and soft dorsal, anal, and caudal fins form simul- taneously with notochord flexion (5.8 mm); branchiostegal rays form and caudal fin round (6.2 mm); incipient anal fin margin partially differentiated (7.4 mm); incipient dorsal fin margin partially differentiated (7.5 mm); spi- nous dorsal fin origin situated over preanal myomere 3-4, soft dorsal origin situated over postanal myomere 16 (7.5 mm); predorsal length 32.3% TL (range: 29.0-44.2% TL); 38.3% SL (range 31.2-53.3% SL; 7.5 mm); in- cipient anal fin margin completely differenti- ated (7.8 mm): pelvic buds form anterior to spinous dorsal fin origin (>8.1 mm); yolk com- pletely absorbed and first pelvic fin rays form (9.3 mm); no swim bladder forms; gut straight; entire finfold absorbed (8.1 mm). Scale for- mation initiated in 15.5 mm juvenile at base of caudal peduncle. Squamation nearly com- plete by 18 mm SL. Scales absent from the cheeks, opercles, nape, breast, prepectoral area, and middle of abdomen. Meristics—Myomere number in E. perc- nurum constant posthatching, preanal myo- meres 15, postanal 19-21 (x = 19.9, n = = 20), 36 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-2) ley 4 a. dorsal, b. lateral views, c. ventral views. with 34—36 total myomeres (* = 34.8, n = 20). Total vertebrae number 33-35 (x = 34.0, n = 5), including one urostylar element (from cleared and stained specimens from Little Riv- er). Scales in the lateral series ranged from 38-48 (« = 43.3; mode = 45; n = 5). Paired Etheostoma percnurum, duskytail darter, newly hatched yolk sac larva, 5.1 mm TL, Little River, Tennessee. and median fin ray values and lengths at ap- pearance are presented in Table 2. Pigmentation.—At 4.9-5.7 mm, newly hatched larvae: body translucent; amber oil globule flattened, located anteriorly in yolk sac; highly branched vitelline plexus (red in EGG AND LarVAL DEVELOPMENT IN Darters—Simon and Layman 37 crete? VE eee ac EG. D. TL larva lateral view. life) over oil globule and anterio-ventral por- tion of yolk sac; melanophores on yolk sac most concentrated ventrally, posterio-laterally, and at body-yolk juncture; prominent patch of melanophores on top of head; melanophores irregularly distributed along dorsum, mid-lat- erally along myosepta, and ventrally toward the tail (Fig. 4A, B). Melanophores developing anteriorly on top of head toward snout (6.1— 6.2 mm; Fig. 4C). Body straw-colored, less translucent; dense patch of stellate melano- phores on top of cranium; orbital bar of me- lanophores developing from opercle to snout: melanophores concentrated medially along dorsum and mid-laterally along horizontal sep- tum; melanophores beginning to develop on o soft dorsal, caudal, and anal fin rays; subcuta- Etheostoma percnurum, duskytail darter, Little River, Tennessee. a. 6.9 mm TL larva lateral view, b. 8.2 mm neous melanophores dorsally on gut (7.4-8.3 mm; Fig. 5). Pre- and post-orbital bars dis- tinct; melanophores forming indistinct blotch- es dorsally (7-8) and mid-laterally; light scat- tering of melanophores on soft dorsal, caudal, and anal rays, and to lesser extent on spinous dorsal fin; subcutaneous melanophores on back of head and opercles; gold iridescent pig- ment in eyes (9.2-9.8 mm). Juveniles pigmen- tation, body opaque, straw-colored, heavily pigmented with melanophores; melanophores on top of head concentrated and confluent; dorsal and lateral blotches wider, diffuse, with indistinct dorso-lateral connections between them; soft dorsal, anal, and caudal fin rays lined with melanophores; few melanophores on pectoral and pelvic fin rays, spinous dorsal 38 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-—2) Fic. 6. 15.5 mm TL juvenile lateral view. fin, underside of head, breast, and belly; gold iridescence in eyes more pronounced (0) The 15.5 mm; Fig. 6). A wild juvenile at 18 mm SL: squamation nearly complete; pigmenta- tion approaching that of adult: body straw-col- ored, 12 lateral vertical brown bars connected dorsolaterally to 6-7 irregular brown dorsal saddles, dark humeral spot, and cheeks speck- led with melanophores. DISCUSSION Species of Catonotus are well known for their derived spawning habit of clustering eggs on the undersides of stones (8, 22). Etheos- toma flabellare spawns beneath large stones in riffles, runs, or pools with slow to moderate current (12, 18). Spawning of E. f. lineolatum occurs in early April until May (23, 24, 25). In Wisconsin and Minnesota, spawning initiated Etheostoma percnurum, duskytail darter, Little River, Tennessee. a. 9.5 mm TL early juvenile lateral view, b. in early May and continued until June at tem- peratures between 13°-17°C, but continued until mid-July when temperatures approached 25°C in Iowa and Illinois (17). The duskytail darter spawns in Little River from late April through June in pools and moderate runs be- neath slab-shaped cobbles (5). Etheostoma f. lineolatum and E. percnurum are sympatric, however, the former species is referred to as E. f. flabellare by McGeehan (2). Duskytail darter larvae are more preco- cious, developing incipient rays in the pectoral fins and complete rays in the median fins ear- lier than striped fantail darters; however, squa- mation begins later, at lengths >15 mm TL. These two taxa can be distinguished from oth- er described members of the E. flabellare spe- cies group, E. f. flabellare and E. kennicotti, based on myomere number, pigmentation and EGG AND LARVAL DEVELOPMENT IN DarTERS—Simon and Layman 39 TabLe 4. Summary comparison of meristic, pigmentation, and ontogenetic event characteristics for four taxa of the Etheostoma flabellare species group. E. flabellare flabellare Characteristic Size and shape Egg diameter 2.2-2.7 mm 2.0-2.8 mm E. flabellare lineolatum E. kennicotti E. percnurum 1.9-2.5 mm 2.6-3.3 mm Hatching length 4.7-6.2 mm 4.5-5.9 mm 4.14.7 mm 4.5-5.4 mm Yolk sac diameter 31% TL 42% TL 32% TL 48% TL Yolk sac absorbed 9-10 mm 7.8-8.0 mm 7.5 mm 9.3 mm Yolk color pale yellow amber orange amber Eye shape oval oval spherical spherical Fin ray formation First pectoral ray (.2—1.5) mm 5.0-5.6 mm Sl 5:2) mm 5.8 mm First dorsal spine 7.2-7.5 mm 5.9-6.2 mm bel 5:2) mm 5.8 mm First soft dorsal ray 7.2-7.5 mm 5.3-5.9 mm 5.1 mm 5.8 mm First pelvic fin ray 8.8 mm §.8-9.2 mm <12.1 mm 9.3 mm First anal ray 7.2-7.5 mm 5.3-5.9 mm 6.9 mm 5.8 mm First caudal ray 7.2-7.5 mm 5.3-5.9 mm 7.5 mm 5.8 mm Morphological event Notochord flexion 7.2 mm 4.9-6.0 mm 6.9 mm 5.8 mm Squamation initiated 13.0 mm 10.0 mm 13.1 mm 15.5 mm Meristics Total myomeres 34-36 34-36 34-35 34-36 Preanal myomeres 15 15 16 15 Postanal myomeres 19-21 (21) 19-21 (19.5) 18-19 (18.5) 19-21 (19.9) fin ray development relative to size (Table 4). Etheostoma kennicotti can be separated from all other members of the E. flabellare species group since it possesses 16 preanal and 18-19 postanal myomeres. All other taxa have 15 preanal and 19-21 postanal myomeres. E. ken- nicotti has melanophores scattered across the yolk-sac similar to E. flabellare, while it differs in possessing a dorsal and ventral cluster pos- terior to the anus. The other three taxa can be separated based on yolk-sac diameter, eye shape, and ontogenetic development of fin rays. The duskytail darter has a spherical eye shape while both subspecies of E. flabellare have an oval eye shape. Significant ditferences exist in the ontogenetic development of fin rays between E. f. flabellare and E. f. lineola- tum. Development of fin rays is more preco- cious in E. f. lineolatum, occurring at smaller length intervals for all but the formation of the first pelvic fin ray. Yolk sac diameter is greatest in duskytail darter (48% TL), followed by E. f. lineolatum (42% TL), and E. f. flabellare (31% TL), while the yolk sac is absorbed at smaller length intervals in E. f. lineolatum and at similar sizes for E. f. flabellare and duskytail darter. Sympatric species of Catonotus are likely to utilize similar slab rock habitat for spawning (3), and thus, their larvae and juveniles may often be collected in the same habitat. Differ- ences between E. f. lineolatum and other de- scribed Catonotus, E. squamiceps and E. smi- thi, enable accurate identification based on myomere counts and pigmentation. Etheosto- ma squamiceps can be separated from E. f lineolatum because the former possesses 16 preanal and 18-19 postanal myomeres (9). Etheostoma smithi has similar myomere counts to E. f. lineolatum but differs in yolk sac diameter, pigmentation, and formation of the rays (10). Yolk sac diameter is smaller (33.5%) than E. f. lineolatum; pigmentation is restricted to the nape, ventral yolk-sac, and mid-ventral postanal myosepta; and fin ray for- mation occurs later than in E. f. lineolatum. ACKNOWLEDGMENTS The authors wish to thank T. Coon, Michi- gan State University; and B. Simon; and K. D. Floyd, Alabama Department of Natural Re- sources, for field assistance in the acquisition of larval and adult brood stock. The initial manuscript was reviewed by R. Wallus, Ten- nessee Valley Authority, and R. L. Mayden, University of Alabama. R. E. Jenkins and N. 40 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) M. Burkhead provided a prepublication copy of the species account for the duskytail darter from “The Freshwater Fishes of Virginia.” II- lustrations of the striped fantail darter were completed by B. Simon, and duskytail darter by S. R. Layman. This work fulfilled portions of Masters of Science degree programs at the University of Tennessee (SRL) and University of Wisconsin-La Crosse (TPS). LITERATURE CITED 1. Page, L. M. 1983. Handbook of darters. TFH Publ., Neptune City, New Jersey. 2. McGeehan, C. M. 1985. Multivariate and univariate analysis of the geographic variation within Etheostoma fla- bellare (Pisces: Percidae) of eastern North America. Un- publ. Ph.D. Dissertation. Ohio State University, Colum- bus. 3. Page, L. M. and D. W. Schemske. 1978. The effect of interspecific competition on the distribution and size of darters of the subgenus Catonotus (Percidae: Etheosto- ma). Copeia 1978:406—-412. 4. Jenkins, R. E. and N. M. Burkhead. 1994. The freshwater fishes of Virginia. American Fisheries Society, Bethesda, Maryland. 5. Layman, S. R. 1991. Life history of the relict dusky- tail darter, Etheostoma (Catonotus) sp., in Little River, Tennessee. Copeia 1991:471-485. 6. Starnes, W. C. and D. A. Etnier. 1980. Fishes. Pp. B1-B134. In D. C. Eagar and R. M. Hatcher (eds.) Ten- nessee’s rare wildlife. Volume I. The vertebrates. Tennes- see Wildl. Res. Agency and Tennessee Depart. Conserv., Nashville. 7. Jenkins, R. E. 1991. Freshwater and marine fishes. Pp. 319-373. In D. W. Linzey (ed.) Endangered and threatened plants and animals of Virginia. Va. Polytech. Inst. St. Univ., Blacksburg. §. Williams, J. E., J. E. Johnson, D. A. Hendrickson, S. Contreras-Balderas, J. D. Williams, M. Navarro-Mendoza, D. E. McAllister, and J. E. Deacon. 1989. Fishes of North America endangered, threatened, or of special con- cern: 1989. Fisheries 14(6):2—20. 9. Page, L. M. 1985. Evolution of reproductive be- haviors in percid fishes. Ill. Nat. Hist. Surv. Bull. 33:275—- 295. 10. Simon, T. P. 1987. Description of eggs, larvae, and early juveniles of the stripetail darter, Etheostoma kenni- cotti (Putnam) and spottail darter, E. squamiceps Jordan (Percidae: Etheostomatini) from tributaries of the Ohio River. Copeia 1987:433-442. 11. Simon, T. P. 1988. Identification of eggs, larvae, and early juveniles of the slabrock darter, Etheostoma smithi, from the Cumberland River drainage, Kentucky. Trans. Ky. Acad. Sci. 49(1/2):15-20. 12. Lake, C. T. 1936. The life history of the fan-tailed darter Catonotus flabellaris flabellaris (Rafinesque). Am. Midl. Nat. 17:117-135. 13. Cooper, J. E. 1979. Description of eggs and larvae of fantail (Etheostoma flabellare) and rainbow (E. caeru- lewm) darters from Lake Erie tributaries. Trans. Am. Fish. Soc. 108:46—56. 14. Auer, N. A. (ed.) 1982. Identification of larval fishes of the Great Lakes basin with emphasis on the Lake Mich- igan drainage. Great Lakes Fishery Commission Spec. Publ. 82-3. 15. Paine, M. D. 1984. Ecological and evolutionary con- sequences of early ontogenies of darters (Etheostomatini). Env. Biol. Fish. 11:97—106. 16. Sumida, B. Y., B. B. Washington, and W. A. Laroche. 1988. Illustrating fish eggs and larvae. Pp. 33-35. In H. G. Moser, W. J. Richards, D. M. Cohen, M. P. Fahay, A W. Kendall, Jr., and S. L. Richardson (eds.) Ontogeny and systematics of fishes. American Society of Ichthyologists and Herpetologists Spec. Publ. 1. 17. Simon, T. P. 1985. Descriptions of larval Percidae inhabiting the upper Mississippi River basin (Osteichthy- es: Etheostomatini). Unpubl. M.S. Thesis. University Wis- consin—La Crosse. 18. Winn, H. E. 1958. Comparative reproductive behav- ior and ecology of fourteen species of darters (Pisces-Per- cidae). Ecol. Monogr. 28(2):155-191. 19. Smith, P. W. 1979. The fishes of Illinois. Univ. Illinois Press, Champaign. 20. Becker, G. C. 1983. Fishes cf Wisconsin. Univ. Wis- consin Press, Madison. 21. Layman, S. R. 1984. The duskytail darter, Etheosto- ma (Catonotus) sp., confirmed as an egg-clusterer. Copeia 1984:992-994. 22. Braasch, M. E. and R. L. Mayden. 1985. Review of the subgenus Catonotus (Percidae) with descriptions of two new darters of the Etheostoma squamiceps species group. Occ. Pap. Mus. Nat. Hist. Univ. Kansas 119. 23, Eddy, S. and J. C. Underhill. 1974. Northern fishes. University of Minnesota Press, Minneapolis. 24. Pflieger, W. L. 1975. The fishes of Missouri. Missouri Department of Conservation, Columbia. 25. Hubbs, C. 1985. Darter reproductive seasons. Co- peia 1985:56-68. Trans. Ky. Acad. Sci., 56(1—2), 1995, 41-46 Aspects of the Dragonfly and Damselfly (Odonata) Community of Buck Creek, Pulaski County, Kentucky RANDALL G. PAYNE! AND GUENTER A. SCHUSTER Department of Biological Sciences, Eastern Kentucky University, Richmond, Kentucky 40475 ABSTRACT Buck Creek is a fifth-order tributary of the upper Cumberland River, in southcentral Kentucky. Thirty two species of Odonata were found to inhabit this stream. Ecological data, particularly seasonality, of 31 species are presented, along with behavioral observations. Published flight seasons were extended for 5 species of Anisoptera and 6 species of Zygoptera for Kentucky. Seasonal life histories are potentially impor- tant in understanding the dynamics involved in niche segregation of a diverse community of general pred- ators. INTRODUCTION A review of the literature concerning the Odonata of Kentucky revealed a paucity of ecological data. Resner (1) presented the lat- est distributional list with seasonality data for most species. The literature review showed that the geographical area of this study had been given little attention in surveys of Odo- nata (1). Additionally, few seasonality studies of communities of lotic Odonata have been published (2-4). We undertook this study to characterize the community of Odonata of Buck Creek, Pulaski County, Kentucky. Because of the desire to observe as many species as possible and offer a complete documentation of seasonality, 2 flight seasons were incorporated in this study (1991 and 1992). Specifically, we wanted to (1) determine the odonate community composi- tion; (2) document specific flight seasons; (3) make ecological observations; and (4) observe behavior. In this paper, data are presented for 31 of the 32 species (5) known to inhabit Buck Creek. Stupby AREA Buck Creek, a fifth-order tributary of the upper Cumberland River, flows southward for 107.2 km, draining 767 km?. This stream is located in southcentral Kentucky (37°10'N, 84°30'/W) and flows primarily within the East- ern Highland Rim subsection of the Interior Low Plateaus Physiographic Province (6). The ' Present address: 711 Underwood Avenue, 405D, Pen- sacola, Florida 32504. 4] surface geology is composed principally of Mississippian age limestone (7). The lower 19% of Buck Creek is inundated by the back waters of Lake Cumberland. This occurred with the completion of Wolf Creek Dam on the Cumberland River in 1951. Buck Creek averaged less than 20 m wide and 2 m deep, but had a maximum width of 150 m and a maximum depth greater than 25 m near its mouth (8). Its gr adient was 1.25 m/km (8) with an estimated mean flow of 11.7 km?/m (9). METHODS Six collecting sites (5) were chosen on the mainstem of Buck Creek. Two sites were vis- ited per week, and a collecting circuit of all sites was completed every 3 weeks. Adult col- lections began in June and continued through October 1991. and from April to mid-Septem- ber 1992. At each site, extensive searches for and observations of adults were made by wad- ing upstream and downstream several 100 m. Physical and biological characteristics of each collection and observation site were made. RESULTS Figures 1 and 2 show the flight seasons of Shoh32 species of Anisoptera and Zygoptera, respectively. The dragonfly species not includ- ed here, Somatochlora linearis, was collected as a larva. The Anisoptera flight season began in mid-April with Basigeschna janata and con- cluded by end of October with Boyeria vinosa (Fig. 1). However, the Zygoptera flight season began in early May with Calopteryx maculata and concluded with Hetaerina americana and Argia translata in October (Fig. 2). Thus, the AESHNIDAE Basiaeschna Janata Boyeria vinosa CORDULIIDAE Epitheca princeps Neurocordulia yamaskanensis GOMPHIDAE Dromogomphus spinosus Gomphus lineatifrons Gomphus lividus Gomphus viridifrons Hagenius brevistylus Stylogomphus albistylus LIBELLULIDAE Libellula luctuosa Libellula lydia Libellula_ pulchella Erythemis simplicicollis Sympetrum vicinum Pachydiplax longipennis MACROMIIDAE Didymops transversa Macromia alleghaniensis EG sale * TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-—2) ie Adult flight seasons of Anisoptera collected and observed (June-November 1991; April-mid-September 1992) at Buck Creek, Pulaski County, Kentucky. Asterisks (*) indicate single date record. adult odonate flight season persisted for 190 known days (Tables 1 and 2). Of the 2 sub- orders, anisopterans had a shorter flight sea- son (Figs. 1, 2). Student's t test was employed to compare the flight season between the 2 suborders (excluding those with less than 7 days recorded flight period). To compute the value of t, the flight season of each species was determined by taking the 2 seasons and aver- aging their flight period. This resulted in a sta- tistically significant (P < 0.05) difference in flight season length. Kentucky flight season range extensions were recorded for 5 species of Anisoptera (Table 1) and 6 species of Zy- goptera (Table 2). DISCUSSION Flight season in this study began with the first observation or collection of a reproduc- tively mature adult. Corbet (10) reported that most reproductively mature Zygoptera live ap- proximately 1 to 2 weeks and may extend to 5 to 8 weeks. Reproductively mature Anisoptera live 2 to 3 weeks and may extend to 3 to 6 weeks (10). Thus, synchrony of emergence was applied to those species with a flight sea- son of approximately 6 weeks or less. Most species collected were typical of lotic habitats; however, the Libellulidae character- istically breed in lentic waters. Westfall (11) reported the large genus Libellula and Pach- ydiplax from lotic waters. The upper one-half of Buck Creek had many braids that became isolated and thus lentic for a significant amount of time during both years of this study. This type of environment was previously re- ported in Buck Creek (7, 12). Pachydiplax lon- gipennis was observed in large numbers at the most upstream site (State Route 70) (5); Li- bellula lydia, L. pulchella and L. luctuosa also were associated with the ponded water at this site. Of the 2 major isolated braids here, P. CALOPTERYGIDAE DRAGONFLIES AND DAMSELFLIES IN KENTUCKY—Payne and Schuster 43 A Calopteryx maculata Hetaerina americana COENAGRIONIDAE Argia fumipennis violacea Argia moesta Argia sedula Argia tibialis Argia translata Enallagma _ basidens Enallagma civile Enallagma divigans Enallagma exsulans Ischnura posita > cae Ischnura_ verticalis J J A Fic. 2. Adult flight seasons of Zygoptera collected and observed (June—November 1991; April-mid-September 1992) at Buck Creek, Pulaski County, Kentucky. Asterisk (*) indicates single date record. TABLE l. Flight seasons for adult Anisoptera based on collections and sightings (June-November 1991; April-mid- September 1992) at Buck Creek, Pulaski County, Kentucky. Addition signs (+) indicate flight season extensions for Kentucky. Aeshnidae Basiaeschna janata (Say) Boyeria vinosa (Say) Corduliidae Epitheca princeps (Hagen) Neurocordulia yamaskanensis Provancher Gomphidae Dromogomphus spinosus Selys Gomphus (Arigomphus) ) lentulus Needham Gomphus (Gomphurus) lineatifrons Calvert Gomphus (Hylogomphus) viridifrons Hine Hagenius brevistylus Selys Stylozomphus albistylus (Hagen) Libellulidae Libellula luctuosa, Burmeister Libellula lydia (Drury) Libellula pulchella Drury Erythemis simplicicollis (Say) Sympetrum vicinum (Hagen) Pachydiplax longipennis (Burmeister) Macromiidae Didymops transversa (Say) Macromia alleghaniensis (Williamson) 20 April +-18 May + 3 Aug.—26 Oct. 5 July-10 Aug. 18 May 31 May-—1 Sept. 1 June + 12 May-31 July 1S May 9 June- 7 Sept. + 1 June—9 June 19 July—3 Aug. 22 xe Aug. 21 June—22 June 14 July—31 Aug. 13 Sept—4 Oct. 19 July—7 Sept. 18 May + 5 July-7 Sept. + 44 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) TaBLE 2. Flight seasons for adult Zygoptera based on collections and sightings (June-November 1991; April- mid-September 1992) at Buck Creek, Pulaski County, Kentucky. Addition signs (+) indicate flight season exten- sions for Kentucky. Calopterygidae Calopteryx maculata (Beauvois) Hetaerina americana (Fabricius) 11 May-13 Sept. + 12 July4 Oct. Coenagrionidae Argia fumipennis violacea Hagen) Argia moesta (Hagen) Argia sedula (Hagen) Argia tibialis Rambur Argia translata Hagen Enallagma basidens Calvert Enallagma civile (Hagen) 13 July Bnalloome divagans Selys 1 June—22 June Enallagma DR ike (Hagen) 18 May-1 Sept. Ischia posita (Hagen) 11 May-13 Sept. + Ischnura verticalis (Say) 12 May-14 July 31 May-13 Sept. + 21 June-13 Sept. + 21 June-31 Aug. 31 May-7 June 31 May +-4 Oct. + 12 May +-13 Sept. longipennis was associated with the smaller one that flowed during high-water conditions. The aeshnids, Basiaeschna janata and Boy- eria vinosa, had dramatically different flight seasons (Fig. 1). Thus, these 2 similarly adapt- ed predators demonstrated temporal segrega- tion in this study. Paulson and Jenner (4) also found B. janata to be synchronized “spring” species (13), while B. vinosa was asynchronous and overwintered in many instars. There was a marked segregation of season- ality among the 2 species of Macromiidae, Di- dymops transversa and Macromia alleghanien- sis (Fig. 1). Although D. transversa was col- lected on one date only in this study (Table 1), it is known to be a synchronous spring flyer (1, 4). Macromia alleghaniensis had an asyn- chronous emergence and flew from mid- to late summer (Fig. 1). Both these species were found in the lower two-thirds of the stream, becoming more common downstream. The family Libellulidae was represented by 6 species; 3 were congeneric (Table 1). While Libellula lydia and Erythemis simplicicollis were collected along most of Buck Creek, the other species had more restricted distribu- tions. Only Sympetrum vicinum had a segre- gated flight season from other libellulids (Fig. 1). Boehms (14) studied the ecology and de- velopment of this species. Observations indi- cated that Pachydiplax longipennis tolerated greater flow velocity than other libellulids. Thus, it occupied a different habitat than the other species. Libellula lydia and Libellula pulchella exhibited interspecific aggression. Both species occurred commonly around a lentic habitat created for flood control. Spatial segregation was observed for L. lydia, which perched on marginal vegetation, and L. pul- chella, which preferred fallen trees and limbs in or near the water. Erythemis simplicicollis was collected or observed primarily along the fifth-order section of the stream. Adult E. sim- plicicollis utilized dense beds of Justicia amer- icana or nearby rocks on which to perch. While the gomphids represented the great- est numbers of individuals collected and ob- served, their flight season was especially inter- esting as there was no segregation between species (Fig. 1). Most members of this family were primarily stream dwellers and relatively little was known concerning the factors gov- erning niche segregation. With many species exhibiting asynchronous emergence, larvae ex- hibited a great spread of taste The mecha- nism(s) Ponte olling their niche segregation must occur during. ae larval life history. Mor- phological carianone in the design of their la- bium may serve significantly in niche segre- gation (15). Also, as sprawlers in leafmats and Burowerss this group occupied the majority of available habitat in the stream (horizontal dis- tribution). Another strategy allowing these cryptic, sedentary odonates to withstand niche overlap may result from slow development which requires long generation times (16). The family Calopterygidae had 2 represen- tatives at Buck Creek, Calopteryx maculata and Hetaerina americana. Calopteryx macu- lata had the earliest flight season of the 2 spe- cies (Table 2). This species demonstrated asynchronous emergence, as evidenced by its long flight season (Fig. 2). Calopteryx macu- lata preferred the more heavily forested sec- tions of the stream, utilizing the dense forest for perches. Hetaerina americana emerged 2 months later and flew until early October (Ta- ble 2). The latter species also emerged asyn- chronously; however, it was observed to oc- cupy a different habitat, principally boulders and exposed gravel associated with riffles. These damselflies were always observed in DRAGONFLIES AND DAMSELFLIES IN KENTUCKY—Payne and Schuster 45 open, sunny situations, as opposed tos ma- culata. The difference in flight season between the 2 species may be due to developmental pat- terns as larvae. Paulson and Jenner (4) found Calopteryx sp. to overwinter in prefinal in- stars, final (F) F-2 and F-1; Hetaerina sp. overwintered in F-4 to F-2 stages. Develop- mental segregation in these 2 morphologically similar species may be a primary mechanism in controlling interspecific competition. Thus, flight-season variations between these spec ies did indicate that larval-size variation is em- ployed as a competitive strategy. Within the large family Coenagr ionidae, long, overlapping flight seasons predominated among the species (Fig. 2). An exception, ou allagma divigans flew from 1 June through 2 June. This species showed synchr onous emer- gence associated with “spring” species (13). Very similar flight-season observations for this species have esi previously demonstrated (4, 17). Paulson and Jenner (4) found E. divigans larvae to overwinter predominantly in °F =i stages in North Carolina. Many studies have reported diverse odonate communities coexisting in relatively small geo- graphic areas (2, 17-19). As general predators, odonates must roid niche overlap by some mechanism(s). It has been suggested that the seasonal segregation of edortates is an impor- tant mechanism to reduce similarly adapted species (15, 16, 20). Van Noordwijk (20) also found spatial separation between 7 genera of an 8-species zygopteran community. Michiels and Dhondt (21) dem- onstrated that 3 species of Sympetrum dis- tinctly partitioned resources both spatially and temporally. Interestingly, Zygoptera had a sta- tistically significant longer flight season than Anisoptera. However, the placement of the longer flight season was not expected. Anisop- tera began flight in early spring, while Zygop- tera did not begin flight until mid-spring. One might expect a competitive advantage gained by damselflies if they had a flight season prior to dragonflies because of fewer potential large predators and the large emergences of poten- tial prey (Chironomidae). This. pattern of flight season had been reported by other studies (17, 99). Detailed life-history studies are needed for many odonates. In addition, more studies are niche overlap of needed to determine the dynamics controlling niche segregation in diverse communities of odonates. ACKNOWLEDGMENTS We thank D. L. Batch and J. R. Maki of Eastern Kentucky University for their sugges- tions and corrections in an earlier manuscript. Our gratitude to S. L. Jones for providing funds and equipment for this study. LITERATURE CITED 1. Resner, P. L. 1970. An annotated check list of the dragonflies and damselflies (Odonata) of Kentucky. Trans. Ky. Acad. Sci. 31:32-44. 2. Bick, G. H. and J. C. Bick. 1958. The ecology of the Odonata at a small creek in southern Oklahoma. J. Tenn. Acad. Sci. 33:240-251. 3. Cross, W. H. 1955. Anisopteran Odonata of the Sa- vannah River Plant, South Carolina. J. Elisha Mitchell Sci. Soc. 719-17. 4. Paulson, K. R. and C. structure in overwintering larval Odonata in North E. Jenner. 1971. Population Caro- lina in relation to adult flight season. Ecology 52:96—-107. 5. Payne, R. G. and G. A. Schuster. 1993. The drag- onflies and damselflies (Odonata) of Buck Creek, Pulaski County, Kentucky. Ent. News 104:165—170. 6. Quarterman, E. and R. L. Powell. 1978. Potential ecological/geographical natural landmarks on the Interior Low Plateau. U.S. Dept. Interior, Washington, IDG. 7. Schuster, G. A., R. S. Butler, and D. H. Stansbery. 1989. A survey of the unionids (Bivalvia: Unionidae) of Buck Creek, Pulaski County, Kentucky. Trans. Ky. Acad. Sci. 50:79-S5. 8. Cicerello, R. R. and R. S. Butler. 1985. Buck Creek, Cumberland River drainage, Kentucky. Brimleyana 11:133-159. 9. USACE. 1976. Water quality conditions in Lake Cumberland. U.S. Army Corps of Engineers, Nashville District, Nashville. 106 pp. 10. Corbet, P. S. 1980. Biology of Odonata. Ann. Rev. Entomol, 25:189-217. 11. Westfall, M. J., Jr. 1984. Odonata. Pp. 126-176. In R. W. Merritt and K. W. Cummins (eds.) An introduction to the aquatic insects of North America, 2nd ed. Kendell/ Hunt Publishing Company, Dubuque, Iowa. 12. Floyd, M. A. and G. A. Schuster. 1990. The cad- disflies (Insecta: Trichoptera) of the Buck Creek System, Pulaski County, Kentucky. Trans. Ky. Acad. Sci. 51:127— 134. 13. Corbet, P. S. 1954. Seasonal regulation in British dragonflies. Nature 174:655. 14. Boehms, C. N. 1971. ture upon embryonic diapause and seasonal regulation in Sympetrum vicinum (Hagen) (Odonata: Libellulidae). Ph.D. Dissertation. Univ. North Carolina, Chapel Hill. Fishes of The influence of tempera- 46 TRANS. KENTUCKY ACADEMY OF SCIENCE 56( 1-2) 15. Corbet, P. S. 1962. A biology of dragonflies. With- erby Ltd., London. 16. Johnson, D. M. and P. H. Crowley. 1980. Odonata “hide-and-seek”: habitat specific rules? Pp. 569-579. In W. C. Kerfoot (ed.) The evolution and ecology of Zoo- plankton populations. Special Symposium 3. American So- ciety of Limnology and Oceanography. Univ. Press of New England, Hanover, New Hampshire. 17. Johnson, D. M., C. C. Coney, and M. J. Westfall, Jr. 1980. The Odonata of Bays Mountain Park, Sullivan County, Tennessee. J. Tenn. Acad. Sci. 55:73-76. 18. Benke, A. C. and S. S. Benke. 1975. Comparative dynamics and life histories of coexisting dragonfly popu- lations. Ecology 56:302-317. 19. Kormondy, E. J. and J. L. Gower. 1965. Life his- tory variations in an association of Odonata. Ecology 46: 882-886. 20. Van Noordwijk, M. 1978. A mark—recapture study of coexisting zygopteran populations. Odonatologica 7: 353-374. 21. Michiels, N. K. and A. A. Dhondt. 1987. Coexis- tence of three Sympetrum species at Den Diel Mol, Bel- gium (Anisoptera: Libellulidae). Odonatologica 16:347— 360. 22. Johannsson, O. E. 1978. Co-existence of larval Zy- goptera common to the Norfolk Broads (U.K.) I. Tem- poral and spatial separation. Oecologia 32:303-321. Trans. Ky. Acad. Sci., 56(1-2), 1995, 47-53 Serological Evidence for Borrelia sp. in Peromyscus leucopus from Western Kentucky ZIXING WANG, LEON DUOBINIS-GRay, STEPHEN WHITE, Ep SNODDyY, AND JAMEs STUART* ‘Department of Biological Sciences, Murray State University, Murray, Kentucky 42071 °OSA 1S 104B, Tennessee Valley Authority, Water Resources, Muscle Shoals, Alabama ABSTRACT During this study, 312 Peromyscus leucopus were surveyed for the presence of Borrelia sp. Mouse serum was screened for antibodies against Borrelia sp. by an immunoblot te chnique using whole spirochete antigen. By this technique, 21 of 312 (6.7%) of the samples appeared to be serologically positive. Each positive sample was further tested by western blotting against B. burgdorferi whole-cell proteins. Eighteen (5.7%) of the 21 samples were confirmed to possess antibodies which bound to the 39/41 kDa protein band on western blots. Equal numbers of positive samples (9 and 9) were obtained from the eastern and western shores of Kentucky Lake. A total of 40 ticks were removed from 24 of 312 mice and examined by the indirect fluorescent antibody (IFA) technique for the presence of spirochetes. All samples were negative by this method. Mouse ear biopsies were collected and incubated in BSK medium, however B. burgdorferi was not recovered by this culturing technique. INTRODUCTION Borrelia burgdorferi is a bacterial spiro- chete which is the etiological agent of Lyme disease. This agent has become tlie most com- mon v eetorhonie pathogen in the United States (7) and is transmitted by ticks. The primary tick vector of B. burgdorferi in the northeastern portion of the United States is Ixodes scapularis (6, 7, 11), whereas I. pa- cificus is most common in the western U.S. While neither of these vectors have been re- ported from Kentucky, it does appear that Lyme disease transmission is occurring in the state. Pelletier et al. (7) reported that 51 Ken- tucky cases of Lyme disease, based upon CDC criteria, met the case definition between 1985 and 1990. This information suggests that either I. sca- pularis is present, but not yet identified, or another vector is transmitting the disease. Re- cent evidence by Teltow et al. (12) in Texas and Luckhart et al. (3) in Alabama indicated that other species of ticks harbor B. burgdor- feri. Among the species noted was the lone star tick, Amblyomma americanum, a common tick in western Kentucky. Levine et al. (2) reported that in the north- eastern United States the white-footed mouse, * Corresponding author. A7 Peromyscus le ae serves as the primary vertebrate host for I. scapularis larvae and nymphs, thus it is the primary reservoir host for B. burgdorferi. In the southeastern U.S., Dermacentor variabilis larvae and nymphs in- fest P. lewcopus most often. Schwan et al. (8) reported that white-footed mice remain per- sistently infected with B. burgdorferi. This in- formation indicated that infected mice should be seropositive for antibodies against B. burg- dorferi long after initial exposure and elevated antibody titers should be detectable weeks or even months after infection. In order to ascertain whether B. burgdorferi possibly exists in western Kentucky, a study was initiated to examine the serum of white- footed mice for antibodies against Borrelia sp. The study was also designed to attempt cul- ture of spirochetes from tissue samples of white-footed mice. Additionally, ticks removed from mice were analyzed by indirect fluores- cent antibody technique (IFA) for the pres- ence of Borrelia sp. MATERIALS AND METHODS Bacterial Strain. —The B. burgdorferi—Guil- ford strain used in this study was obtained from Dr. Gary Mullin, Dept. of Entomology, Auburn University, Alabama 36849. The Guil- ford strain is a high-passage isolate of B. burg- 48 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) positive G ] positive KENTUCKY LAKE [D) / positives nS ZS E TENNESSEE 1 positive positive \ ! | [| 3 e: positives dy Localities of P leucopus from LBL and the western shore of Kentucky Lake. Fic. 1. Sample localities from LBL and TVA recreation areas: a total of 200 P. leucopus were collected from TVA public use areas on the western shore of Kentucky Lake: B (Jonathan Creek), C (White Sands Beach), D (Wildcat Creek), and E (Blood River). The positive samples were found in localities C, D, and E. A total of 112 P. leucopus were collected from the eastern shore of Kentucky Lake (LBL areas A, F, G, H, and I). Mice with positive sera were found in each of these five localities. dorferi originally isolated by Alan Steere from Ixodes in Guilford, Connecticut. Animal Capture.—Between 30 June 1990, and 29 August 1991, 185 P. leucopus were live- trapped at 4 sites along the western shore of Kentucky Lake in Calloway and Marshall counties, Kentucky. The 4 sites (Fig. 1), Blood River (BR), Wildcat Beach (WC), White Sands Beach (WS), and the Boy Scout Camp on Jonathan Creek (JC) were selected as areas of high human activity and potential human exposure to B. burgdorferi. Each site included campgrounds, beaches, picnic areas, and/or natural areas and trails that concentrate rec- reational activities on TVA-owned lands. Ad- ditionally, 127 mice from the eastern shore of BORRELIA IN KENTUCKY PEROMYSCUS—Wang et al. 49 Kentucky lake at Land Between the Lakes (LBL) recreation area were examined (Fig. 1). Sherman live traps baited with peanut but- ter or bird seed were set and checked daily. Captured animals were transported to labo- ratory facilities at Murray State University. Animal Handling.—Following transport, mice were held for a period of at least 3 days over water-filled containers lined with double- sided tape in order to collect engorged ticks. Any ticks collected were placed ate fale and held at 4°C for analysis by IFA (see below). At the end of the holding period, animals were sacrificed by etherizing, bled, and ears re- moved. Blood was allowed to clot, and serum was collected, frozen at —80°C, and analyzed by immunoblotting. Ear punch biopsies were performed on each mouse as described below. Ear tissue remaining from the mice was stored frozen at —80°C. Cultivation of Borrelia burgdorferi—The medium for cultivation was the standard Bar- bour-Stoenner-Kelly (BSK) described by Bar- bour (1). The ear-punch biopsy method of Sin- sky and Piesman (10) was initially used to cul- ture spirochetes from captured wild mice. During the study this method was modified as follows. A pie- -shaped wedge was clipped from the ear and soaked in 10% bleach for 10 min followed by a 10-min soak in 70% ethanol. The tissue was minced, added to 100 pl of BSK ina microcentrifuge tube and incubated at 32°C for 2 weeks in a candle jar. The culture was checked twice (7 days and 14 days) under a phase contrast microscope for the presence of spirochetes. With each new batch of media, positive controls were cultured by inoculating fresh media with a 10 pl aliquot of viable spi- rochetes (Guilford) and incubating at 32°C. Indirect Fluorescent Antibody (IFA) Test- ing.—This test was modified from a procedure described by Luckhart et al. (3). Ticks were dissected, and the contents mixed with one drop of sterile water on the surface of a clean slide. This was allowed to air dry, and fixed by mild heating. Fifty wl of polyclonal rabbit anti- B. burgdorferi serum (1/50) was added to the tick smear, and the slide was incubated for 30 min at 37°C in a moist chamber. The slide was washed once in phosphate buffered saline (PBS) for 10 min and air dried. To the dried slide was added 1 drop of goat anti-rabbit gamma globulin (1:50 dilution) labeled with fluorescein isothiocyanate (Sigma, St. Louis, Missouri). Incubation procee ide d for 30 min at 37°C in a moist chamber. The washed once with PBS as above. with distilled water. The slide was air dried and observed using a fluorescence micro- scope. Positive controls consisted of placing a drop of live spirochetes from BSK culture on a slide instead of a tick smear. Immunoblot Screen.—B. burgdorferi cells were grown into late log phase in BSK medi- um, centrifuged, and washed 3 times in PBS. Cells were resuspended in PBS at approxi- mately 10° cells/ml, and 100 pl aliquots were spotted onto a nitrocellulose sheet held by a ’acudot (American Bionetics: Hayward, Cali- fornia) apparatus. The nitrocellulose was dried, and each spot was cut from the sheet and stored at —80°C. The immunoblot pro- cedure was adapted from Towbin (13). Each dot blot was blocked by incubating for | hr at room temperature (RT) in a solution of 3% bovine serum albumin (BSA, Sigma) suspend- ed in PBS. The blots were then probed with a 1:200 dilution of serum from each experi- mental mouse for | hr at RT. Blots were sub- jected to 2 10 min washes with 1% BSA in PBS, then incubated with a 1:1,000 dilution of peroxidase conjugated goat anti-mouse IgG (H + L) (Jackson Tmiiporcecarelr West Grove, Pennsylvania) suspended in 3% BSA- PBS for 1 hr at RT. Each dot blot was washed 3 times (10 min each) in 3% BSA-PBS then developed with a solution containing o-dian- isidine (25 g/ml) and 0.01% H,O, in 10 mM Tris (pH 7.4) for 10-15 min. Positive and neg- ative control sera were included with each test run. Positive serum was obtained by injecting each mouse in a group of P. leucopus twice with approximately 10° freshly grown. spiro- chetes. One week following the second injec- tion, mice were bled and the serum collected and titrated by the above method. Positive control mouse serum attained a titer in excess of 1:3,200 with the immunoblot method. Neg- ative controls were selected from experimental animals which appeared negative on the initial immunoblot screening. Further titrations of these sera were made at dilutions of less than 1:200 to insure that no antibody was present. Western Blotting —Western blotting was used to confirm eaah positive serum sample from the immunoblotting screen. Whole cell slide was and rinsed 50 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) immunoblot Hies2s experimental serum taken from mice captured at Jonathan Creek; Ws 7 captured at White Sands Beach. proteins were extracted from B. burgdorferi using the boiling water bath method described by Simpson et al. (9). An aliquot containing 275 wg of protein was added to an SDS gel and subjected to polyacrylamide gel electro- phoresis for 4 hr at a constant current of 40 ma. An aliquot containing 50 wg of marker protein (SDS-6; Sigma) was loaded adjacent to bacterial proteins in order to obtain a refer- ence of molecular masses. Following resolu- tion by SDS-PAGE, proteins were electroblot- ted onto nitrocellulose membranes (0.45 pw) (MFS; Dublin, California) using a Bio-Rad (Richmond, California) Transblot Electropho- retic Cell according to manufacturer’s instruc- tions. The transferred membranes were dried Immunoblot screen: each serum sample was diluted 1/200 and used to probe whole spirochete antigen spotted on nitrocellulose. Arrows indicate positive reactivity of experimental serum with B. burgdorferi antigen. C+ and C— = positive and negative serum samples; L1, 10, 11, 83, 102 = experimental serum taken from mice captured from Land Between the Lakes; WC 2, 4, 16, 20, 21, 22, 23, 35 = experimental serum taken from mice captured from Wildcat Beach; Br 1, 6, 7 = experimental serum taken from mice captured from Blood River; Jc 26, 27, 28 = . 8, 9 = experimental serum taken from mice and cut into 3 mm strips for probing with ex- perimental mouse serum. The probing pro- cedure was the same as described above in the immunoblot technique, again including posi- tive and negative control sera. RESULTS Cultivation of Borrelia from Wild Animal Hosts.—All attempts to cultivate B. burgdor- feri from Peromyscus tissue were negative, whereas positive control cultures produced high concentrations of viable spirochetes. Indirect Fluorescent Antibody Testing.—A total of 40 ticks (all D. variabilis larvae or nymphs) were collected from 24 P. leucopus BORRELIA IN KENTUCKY TaBLe 1. Immunoblot screen of Peromyscus serum. Number positive/total number Month* collected Sample site 14 5-8 9-12 Subtotal LBL* 0/5 §/109 2/13 10/127 BR? 1/27 0/10 0/2 1/39 Jc 0/15 0/20 0/15 0/50 WC! 0/5 9/43 — 9/45 WSs 1/5 0/43 — 1/48 Subtotal 2/57 17/227 2/30 Total DVB (6.7%) * Numbers indicate calendar month “Land Between the Lakes » Blood River. © Jonathan Creek. ‘Wildcat Beach. * White Sands during the course of the study. IFA tests were considered negative for all ticks tested. Immunoblot Scr ven.—Using an immuno- blot technique, serum from each mouse was diluted 1:200 and used to probe a whole spi- rochete antigen preparation spotted on nitro- cellulose. Figure 2 illustrates a typical test run with a positive and negative serum control. The positive experimental sera are marked with arrows. Table 1 represents the prelimi- nary results of immunoblot screening by sam- ple site, and approximate time period of the year. Western Blotting —Each positive sample Spun jhe taimnte ge seteee eS subjected to further analysis by western blotting. Serum samples which possessed specific antibodies to the 39/41 kDa protein of B. burgdorferi were considered confirmed positives, since these proteins have been shown to occur in all va- rieties of B. burgdorferi (8, 9, 14) and to spe- cifically mark this species. Figure 3 shows a selection of the confirmation experiments. Each of the 9 experimental serum samples shown in Figure 3 as well as 9 others (not shown) were found to contain antibodies which reacted with the 39/41 kDa protein band. Some additional reactive antibodies were noted, for example: serum sample WC19 (D) reacted with a 56 kDa protein band and LBL100 (H) reacted with 49, 56, and 72 kDa protein bands. The positive serum control pos- sessed antibodies which recognized protein bands at 21, 32) 36; 41, 49, 56, and 72 kDa; whereas the negative serum control failed to PEROMYSCUS—Wang et al. 51 bind any Borrelia proteins. The ge ographic lo- cality of each confirmed positive animal from all localities sampled is illustrated on Figure 1. DISCUSSION The inability to cultivate infectious spiro- chetes in this study was disappointing, but not unusual. Luckhart et al. (3) presented evi- dence of Borrelia sp. in tick populations by the indirect fluorescent antibody technique, but were unable to culture the bacterial agent from a large sample of wild rodents. Also, the inability to demonstrate ticks infected with Borrelia in our study is not unusual given the small sample size, and the fact that we were unable to collect ticks from any animals which were seropositive for this bacterium. The dot blot rapid screen test (Fig. 2) proved to be a reliable indicator of the pres- ence of antibodies versus B. burgdorferi, since 18 of 21 samples were confirmed by western blot analysis. From Table | it is notable that most seropositive animals were collected dur- ing the summer months from Wildcat Beach and 2 localities within LBL (Fig. 1). The west- ern blot test (Fig. 3) showed 18 of 21 positive sera which contained antibodies reactive with the 39/41 kDa band. The 41 kDa protein band has been described by other investigators (8, 14), and corresponds to the flagellin antigen of B. burgdorferi. The 39 kDa protein band has more recently been described by Simpson et al. (9), and was described by these authors as a marker for infection of animals naturally inoculated with B. burgdorferi. It should be noted that these protein bands were resolved on PAGE gels, but the resolution did not ex- tend to nitrocellulose blots. The positive se- rum control lane (C) in Figure 3 displays a broad band of reactivity in the 40 kDa region which includes both 39/41 kDa bands in the PAGE control (lane A). All of the confirmed positives showed this broad band of reactivity which is especially ev- ident from samples in lanes F, G, and L. These findings are in accord with other researchers in the southeastern U.S. who have performed serological surveys of wild animal populations to detect antibodies versus B. burgdorferi. Magnerelli et al. (4) reported that in Con- necticut and other states on the eastern sea- board 51% and 20% of the sera examined (by ELISA) from Odocoileus virginianus and Pro- Sy) TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1 Westernblot Zi ACBC.D EF sGaia: | Western blotting: selected positive serum samples from the immunoblot screen were used to probe whole cell Fic. 3. J Kae proteins of B. burgdorferi subjected to PAGE and blotted to nitrocellulose. Lane A—whole cell B. burgdorferi proteins resolved by SDS-PAGE; B—negative P. leucopus control serum; C positive P. lewcopus control serum; D—L—im- munoblot strips probed with serum collected from mice WC19 (D),WS1 (E),WC35 (F),BR16 (G),LBL100 (H),WC45 (I), WC47 (J),LBL6 (K), and LBL7 (L). Arrows indicate reactive proteins of 39/41 kDa, 56 kDa, and 72 kDa in the control lane. cyon lotor, respectively, contained antibodies to B. burgdorferi. In another ELISA study by Magnarelli et al. (5), several localities in the eastern and southeastern U.S. were surveyed for anti-Borrelia antibodies in the serum of ro- dents. Positive Peromyscus gossypinus sera were obtained from each of the 7 southeastern states surveyed. The rates of positivity ranged from a high of 38% in South Carolina to a low of 15% in Alabama. From a total of 535 P gossypinus serum samples tested, 27% showed antibodies to B. burgdorferi. We con- clude that preliminary evidence from our study indicates a low level of infectivity by Borrelia sp. exists among P. leucopus popula- tions located in western Kentucky. Further studies will focus on identification of which tick species may harbor B. burgdorferi in western Kentucky and document the preva- lence of the bacterium within potential vector populations. ACKNOWLEDGMENTS We thank Gary Mullins and Shirley Luck- hart for valuable information and advice which was necessary to begin this project and Steve Bloemer for invaluable assistance in the trap- ping efforts during this study. We also thank Carl Wood for his expertise in graphics design. This work was supported by grant #TV80544V from the Tennessee Valley Au- thority and a grant from the Committee on BORRELIA IN KENTUCKY PEROMYSCUS—Wang et al. 53 Institutional Studies and Research at Murray State University. LITERATURE CITED l. Barbour, A. G. 1984. Lyme disease spirochetes. Yale J. Biol. Med. 57:71-75. 2. Levine, J. F., M. Wilson, and A. Spielman. 1985. Mice as reservoirs of the Lyme disease spirochete. Am. J. Trop. Med. and Hyg. 34:355-360. 3. Luckhart, S., G. R. Mullen, and J. C. Wright. 1991. Etiologic agent of Lyme Disease, B. burgdorferi detected in ticks (Ascari: Ixodidae) collected at a focus in Alabama. J. Med. Entomol. 28:654—657. 4. Magnerelli, L. A., J. H. Oliver, Jr., H. J. Hucheson, and J. F. Anderson. 1991. Antibodies to Borrelia burg- dorferi in deer and racoons. J. Wildl. Dis. 27:562-568. 5. Magnerelli, L. A., J. H. Oliver, Jr, H. J. Hucheson, J. L. Boone, and J. F. Anderson. 1992. Antibodies to Bor- relia burgdorferi in rodents in the eastern and southern United States. J. Clin. Microbiol. 30:1449-1452. 6. Oliver, J. H., Jr, M. R. Owsley, H. J. Hutcheson, A. M. James, C. Chen, W. S. Irby, E. M. Dotson, and D. Kk. Mclain. 1993. Conspecificity of the ticks Ixodes scapularis and I. dammini (Acari: Ixodidae). J. Med. Entomol. 30: 54-63. 7. Pelletier, A. R., R. F. Finger, and D. M. Sosin. 1991. The epidemiology of Lyme disease in Kentucky. Ky. Ep- idem. Notes Rep. 26:1—3. Isolation and cultivation of 8. Schwan, T. G., K. K. Kime, M. E. Schrumpf, J. E. Coe, and W. J. Simpson. 1989. white-footed mice (Peromyscus leucopus) experimentally infected with the Lyme disease spirochete (Borrelia burg- dorferi). Infect. Immun. 57:3445-3451. 9. Simpson, W. J., W. Burgdorfer, M. E. Schrumpf, R. H. Karstens, and T. G. Schwan. 1991. Antibody to a 39- kilodalton Borrelia burgdorferi antigen (P39) as a marker for infection in experimentally and naturally inoculated animals. J. Clin. Microbiol. 29:236-243. 10. Sinsky, R. J. and J. Piesman. 1989. Ear punch bi- opsy method for detection and isolation of Borrelia burg- dorferi from rodents. J. Clin. Microbiol. 27:1723-1727. 11. Spielman, A., M. L. Wilson, J. F. Levine, and J. Piesman. 1985. Ecology of Ixodes dammini-borne human babesiosis and Lyme disease. Ann. Rev. Entomol. 30:439— 460. 12. Teltow, G. J., P. V. Fournier, and J. A. Rawlings. 1991. Isolation of Borrelia burgdorferi from arthropods collected in Texas. Am. J. Trop. Med. Hyg. 44:469-474. 13. Towbin, H. T. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Nat. Acad. Sci. 76:4350-4354. 14. Wallich, R., S. E. Motsi, M. M. Simon, K. Ebnet, A. Heiberger, and a D. Kramer. 1990. The Borrelia burgdorferi: flagellum-associated 41-kilodalton antigen (flagellin): molecular cloning, expression, and amplifica- tion of the gene. Infect. Immun. 58:1711—1719. Antibody response in Trans. Ky. Acad. Sci., 56(1—2), 1995, 54-56 Pennsylvanian Sharks from Kentucky JAMES X. CORGAN Austin Peay State University, Clarksville, Tennessee 37044 ABSTRACT There are published records of 2 Pennsylvanian sharks from Kentucky that have been identified to species. Edestus minor Newberry, 1866 occurs in the Carbondale Formation of Muhlenberg County and Symmorium reniforme Cope, 1893 is known from the Kendrick Shale of Floyd County. This report adds a third record. Edestus heinrichii Newberry and Worthen, 1870 is reported from the Carbondale Formation of Union County. INTRODUCTION Part of the lower jaw of a shark, with 7 teeth in place, was found during mining in the Hamilton #2 mine of the Island Creek Coal Company, near Uniontown, Union County, Kentucky (Fig. 1). It occurred, at a depth of 350 feet, on a bedding plane that separates the Number 9 Coal from an overlying shale hori- zon within the Carbondale Formation of the Allegheny Series. The jaw fell from the mine roof while a crew was working on shaft main- tenance. Because it fell, some teeth are chipped. Breakage does not obscure basic morphology. The discovery was made in the fall of 1991. Some 2 years later the fossil was taken to the Evansville Museum of Arts and Sciences. At present the specimen belongs to the miner who made the find, Mr. Thomas Nass. He may donate it to the museum. For access to this specimen, I am indebted to Mr. Nass and to Mitchell Lumen, Curator of Collections in the Evansville museum. Edestid Dentition and Classification The arched tooth row, the morphology of teeth, and the appearance of the preserved cartilage identify this Union County fossil as part of the lower dentition of an edestid shark. Sharks of the Order Edestida are reasonably well-known (1). In life, the teeth of the lower jaw were supported by a massive cartilage complex formed by a fusion of the left and right mandibles (2). Mandibles fuse at the symphysis, the zone of juncture between the halves of the jaw. As the shark grew, the sym- physis was produced into a long tooth-bearing rod. Curvature of this cartilaginous support is almost universal in edestids. 54 The symphysial dental battery of the lower jaw matched a comparable zone, the rostrum, in the upper jaw. The rostrum also bore teeth, occluding with those in the lower jaw. The skeletal support for rostral teeth was not stur- dy. Other portions of the skeleton are little mineralized. Thus, edestid species are primar- ily known from symphysial teeth. Shape, size, and dental sculpture place the specimen illustrated in Figure 1 within the ge- nus Edestus Leidy. Edestus ranges from Mis- sissippian to Pennsylvanian (1) and occurs throughout the world. Branson (3) clarified the distribution and synonymy of the type spe- cies. He also evaluated related genera and provided an inventory of species. About 6 valid species of Pennsylvanian Edestus are known from North America. Symphysial dentition is an adequate base for recognizing species. Kentucky Shark Species The shark shown in Figure | is Edestus heinrichii Newberry and Worthen (1870). In the plate explanation which was part of the original description, the name was given as E. henreichsii. This incorrect spelling is just a slip of the pen. It has never caused confusion. When the species was redescribed by New- berry (4), only the name E. heinrichii was used. The name honors a coal mine owner, John P. Heinrich. The type specimen came from his mine in the Pennsylvanian of Bell- ville, Illinois. The original description and the redescription were influenced by other speci- mens, collected in Illinois and Indiana. The only prior record of Edestus in Ken- tucky is from a coal mine near Beech Creek, Muhlenberg County. Jillson (5) provides a well-illustrated, lengthy discussion. The spe- cies is Edestus minor Newberry, 1866. FOSSIL Fic. 1. Edestus heinrichii Newberry and Worton, SHARKS IN KENTU rere ere te ei PTT ATTEN NAT i 6 re 8 9) iI 7 1870, from the Carbondale Formation at the 350-foot level within the Hamilton #2 Mine of the Island Creek Coal Company, near Uniontown, Union County, Kentucky. The specimen is 23.2 cm long, with a maximum height of 6.0 cm. Length of teeth at the junction with cartilage ranges from 3.5 to 4.5 cm. Well-preserved teeth have 11 anterior denticulations and exceeded 2.1 cm in he ight. Breakage prevents den- ticulation counts and height measurement for most teeth. The anterior direction is to the: right. Massive tissue below the teeth is poorly preserved cartilage of the fused mandibles. Kentucky occurrences of E. heinrichii and E. minor mark distributional limits. In the last century, Newberry (4) realized that no species of Edestus was known from Ohio, Pennsylva- nia, or areas to the east. He did not know ‘why and did not comment on distribution to the south, probably because few Pennsylvanian sharks were known from the southern states. More than a century later, there are still no records of edestiids from Ohio, the eastern United States, or adjoining Canadian prov- inces. Today, the southern distribution of Late Paleozoic vertebrates is better known. There is reasonable data on vertebrates from exten- sive Paleozoic outcrops in Tennessee and Al- abama (6, 7). A few vertebrates are known from a relatively small region of Paleozoic out- crops in Mississippi. Those described during the Twentieth Century are well-known (8). Like Ohio and regions to the east, the area south of Kentucky seems to lack edestids. As in Newberry’s day, the absence remains unex- plained. There is one other record of a specifically identified Pennsylvanian shark from Kentucky. What is preserved is a series of punctures within the shell of a marine invertebrate, a nautiloid cephalopod. Punctures are clearly bite marks left by a large marine predator. Mapes and Hansen (9) sticbuted the marks to Symmorium reniforme Cope, 1893, a shark of the Order Cladodontida. Through the work of Williams (10), the anatomy of this shark is almost singularly well- known. Thus species-level bite-mark identifi- cation seems reasonable. The cephalopod that preserves the marks of S. reniforme is from the Kendrick Shale of the Kanawha Series of Eastern Kentuc ky. It was collected near Ligon, Floyd County. The Kendrick Shale is slightly older than beds that yield Edestus in Western Kentucky. Like Kentucky species of Edestus, Sore niforme occurs in Illinois but not in states to the south. The eastern distribution of S. reniforme was not investigated in this study. 56 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-—2) SUMMARY Two species of the shark genus Edestus oc- cur in Pennsylvanian strata of Western Ken- tucky. Identification of both E. minor and E. heinrichii is based on symphysial dentition with attached cartilage. Each species is known from a single specimen. By coincidence, in each specimen seven teeth are present. Both species occur in the same stratigraphic setting. They were found at the interface between the Number 9 Coal and overlying shales of the Carbondale Formation. One species of the Pennsylvanian shark genus Symmorium is known from the Kendrick Shale of Eastern Kentucky. Identification of Symmorium reni- forme is based upon an interpretation of bite marks in a marine invertebrate shell. All spe- cies of Pennsylvanian sharks known from Ken- tucky also occur in states to the north of Ken- tucky. None occurs in reasonably diverse fau- nas from Tennessee and Alabama or in faunas from Mississippi that were described in the Twentieth Century. The eastern distribution of Symmorium was not examined in this study. Species of Edestus do not occur to the east. The cause of this distributional pattern in Pa- leozoic sharks is not known. LITERATURE CITED 1. Moy-Thomas, J. A. and R. S. Miles. 1971. Palaeo- zoic fishes, 2nd ed. Chapman and Hall, Ltd., London. 2. Eaton, Th. H. Jr. 1962. Teeth of edestid sharks. Univ. Kansas Pub. 12:347-362. 3. Branson, C. C. 1963. Type species of Edestus Leidy. Okla. Geol. Notes 23:275-280. 4. Newberry, A. S. 1889. Paleozoic fishes of North America. U.S. Geol. Surv. Mongraph 16:1-340. 5. Jillson, W. R. 1949. Edestus minor, an extinct shark from the Allegheny Series of western Kentucky. Roberts Printing Co., Frankfort, Kentucky. 6. Corgan, J. X. 1976. Vertebrate fossils of Tennessee. Tenn. Div. Geol. Bull. 77:1—100. 7. Thurmond, J. T. and D. E. Jones. 1981. Fossil ver- tebrates of Alabama. Univ. Alabama Press. Tuscaloosa. 8. Daly, E. 1993. A list, bibliography, and index of the fossil vertebrates of Mississippi. Miss. Off. Geol. Bull. 128: 1-48. 9. Mapes, R. H. and M. C. Hansen. 1984. Pennsyl- vanian shark-cephalopod predation: a case study. Lethaia 17:175-183. 10. Williams, M. E. 1985. The “cladodont level” sharks of the Pennsylvania black shales of central North America. Palaeontographica Abt. A 190:83-158. Trans. Ky. Acad. Sci., 56(1—2), 1995, 57 NOTE New records of slender madtom, Noturus exilis Nelson, South Fork Licking River, Pendleton Coun- ty, Kentucky.—As part of a stream investigation by the Kentucky Department of Fish and Wildlife Resources, Di- vision of Fisheries, seine sampling a limestone rubble-rif- fle area, intermixed with gravel and numerous weathered freshwater mussel shells, in the South Fork Licking River below Hayes Station Rd. bridge crossing, 1.5 air km SW of Falmouth (6.9 stream km upstream of confluence with Licking River) 19 August 91, revealed several species of fish: 3 of which were Notwrus. One was identified as No- turus flavus, the other 2 being problematical, having equal jaws and tooth pads on the upper jaw without long distinct backward extensions, were sent to R. R. Cicerello (Ken- tucky State Nature Preserves Commission) who tentative- ly identified them as Noturus exilis, and subsequently sent the specimens to B. M. Burr (Southern Illinois University at Carbondale). In return correspondence, Burr stated “I counted 17-18 anal rays, about 45 caudal rays, 10 POM pores, | internasal pore, 9 pelvic rays, and equal jaws. All of the counts fit N. exilis and there are very few madtoms with equal jaws. Even in weakly pigmented N. exilis there is usually a black marginal band left in the dorsal fin. Un- fortunately neither of these specimens have the band. Other than what I have noted as discrepancies I see no compelling reason not to call them N. exilis.” Based on characters examined, these specimens appear to be more closely allied to N. exilis than any other spe- cies, with noted exceptions, but could possibly be an un- described form. The author wishes to advise collectors to look closely at any suspect N. flavus collected. Noturus exilis is listed as endangered in Kentucky (Warren et al., Trans. Ky Acad. Sci. 47:83-98, 1986) and is known from ten localities only in the state (Burr and Warren, Ky Nat. Pres. Comm. Sci. Tech. Ser. 4, 1986). Voucher specimens of N. exilis are housed at Southern Illinois University at Carbondale (SIUC 19120). I thank Ronald R. Cicerello, Kentucky State Nature Preserves Commission; Brooks M. Burr, Southern Illinois University at Carbondale; and Al Surmont and Fred Howes, Kentucky Department of Fish and Wildlife Re- sources.—Lewis E. Kornman, Kentucky Department of Fish and Wildlife Resources, Division of Fisheries, More- head, Kentucky 40351. Trans. Ky. Acad. Sci., 56(1—2), 1995, 58-61 ACADEMY AFFAIRS THE EIGHTIETH ANNUAL MEETING OF THE KENTUCKY ACADEMY OF SCIENCE The Executive Inn, Paducah, Kentucky 3-5 November 1994 MINUTES OF THE GOVERNING BOARD MEETING KENTUCKY ACADEMY OF SCIENCE Present: Presiding—Larry P. Elliott; Other Governing Board Members: Branley A. Branson, William Bryant, Ju- lia Carter, Robert Creek, Gerald DeMoss, Vincent A. DiNoto, Jr., Patricia K. Doolin, Val Dunham, Blaine R. Ferrell, Valena Hurt, J. G. Rodriguez. Guests in attendance: Jim Meeks, Local Arrangements Committee Chairperson, Andy Aliger, Tennessee Acade- my of Science representative, Chuck Cox, Laura Walker, and Jon Wells, representatives of Citizens Bank and Trust. * The meeting was opened at 1:08 p.m., CST. * President Larry Elliott welcomed participants to the meeting. The meeting was turned over to the represen- tatives from Citizens Bank and Trust for an update on the Marcia Athey Trust Fund (Appendix A). They also discussed their investment strategy. A short discussion followed. It was learned that the bank charges 0.3% of the principal and 6% of the interest income to manage the money in accordance with Kentucky revised stat- utes. Larry Elliott thanked them for their report and work on the trust fund. The minutes of the 27 August 1994 meeting of the Gov- erning Board held in Paducah at the Executive Inn were approved with one correction. Under the Vice Presi- dent's report, it should read “William Bryant” not “Larry Elliott.” Robert Creek presented a short report concerning the 1994 annual meeting (Appendix B). He noted several changes in room assignments and indicated that he had been in contact with the sectional chairs. Robert also discussed a correspondence from the West Virginia Academy of Science regarding a compendium of all state academies publications to be distributed on CD- ROM. It was decided that this correspondence, received by a representative of each university, should be for- warded to each school’s librarian. President's Report Larry Elliott summarized the president's report (Appen- dix C). Awards The winners of the 1994 KAS awards were: Distinguished Scientist—Dr. Donald T. Frazier, De- partment of Physiology and Biophysics, University of Kentucky. 58 * Industrial Scientist—Dr. Fred L. Tungate, United Cat- alysts, Inc., Louisville. * Outstanding College—University Science Teacher—Dr. Barbara A. Ramey, Department of Biological Sciences, Eastern Kentucky University. * Outstanding Secondary Science Teacher—Beverly Lynn White, North Laurel High School. Future Meetings The dates for the joint KAS-TAS meeting is tentatively set for the dates 15-17 November and to be held at Western Kentucky University. Treasurers Report Julia Carter presented the financial status of the KAS (Appendix D). Elections The results of the election of KAS officers are as fol- lows: * Vice-President—Marcus T. McEllistrem, Physics, Uni- versity of Kentucky. * Secretary—Peter X. Armendarez, Chemistry, Brescia College. ¢ Representative to the Governing Board—Social Sci- James F. Hopwood, Anthropology, Northern Kentucky University. Representative to the Governing Board—Physical Sci- Chemistry, Ashland Petroleum Research and Development. ence ences Newsletter Editor Vince DiNoto indicated that he would obtain back issues of the Transactions of the Kentucky Academy of Science at the University of Louisville, store them in the Jeffer- son Community College, SW library on a short term basis in return for one copy of each issue being placed in this library permanently. Robert Creek indicated that ten copies of each issue stored at Eastern Kentucky Uni- versity (nine year period) would be given to Rod Rod- riguez and the remainder would go to Vince DiNoto to be discarded. Transactions Editor Branley Branson pleaded for cooperation in getting the records of Academy affairs (e.g., meeting minutes, var- ious reports, etc.) to him in a timely manner. He indi- cated that he and Varley Wiedemann, who prepares the ACADEMY AFFAIRS annual index, would be resigning from their positions within a year or two. KJAS Director Val Dunham indicated that he was searching for a pro- fessional person to design a color brochure for the KJAS. The KJAS will be sending two students and a chaperon to the AAAS meeting in Atlanta. He men- tioned that there has been a problem with KJAS student paper winners’ attendance at the annual KAS meeting. Executive Secretary's Report Rod Rodriguez commented that the KAS should have the Transactions put on microfiche. Robert Creek asked Rod whether or not copies of the Transactions were still being sent to the archivist, Charles Hayes, at Eastern Kentucky University. Rod said no. Rod commented that something should be done about the library subscription at the University of Louisville. Their request was down from 90 to 70 copies. Rod sug- gested that KAS should terminate its arrangement with the University of Louisville library. Robert Creek asked Rod to bring a specific recommendation before the board at its January meeting. He indicated that the KSTC membership was up for renewal. Last year KAS paid the KSTC $250.00 and they paid KAS $100.00. William Bryant moved that KAS join KSTC again this year, and Pat Doolin seconded. The motion passed. 59 Executive Committee Meeting The meeting came to order at 4:15 p.m., CST. Dates for the joint KAS-TAS meeting were discussed. The 2-4 November date had a conflict with the KSTA meeting, and 15-17 November had a conflict with the NABT meeting. Estimates for the number of papers ran approximately equal to 150 for TOS and 300 for KAS. Business Meeting November 5, 1994 10:00 a.m. President Larry Elliott spoke about the plans for next year's joint meeting with TAS. The meeting will be held at Bowling Green. 3 Treasurer Julia Carter presented a brief financial report and reported on ihe start of an investment bond ladder in August. The interest rate will be a lot better than investment in CDs. Two resolutions were passed: 1. Resolution thanking individuals of Paducah Com- munity College and Murray State University for their hosting of the annual KAS Meeting. 2. Resolution by KAS support for KERA. Both resolutions were passed by the membership. The constitutional changes proposed by Larry Giesman were presented to the membership and passed. Larry Elliott passed on the gavel to President-elect Rob- ert Creek. APPENDIX A Raymond H. Athey Kentucky Academy of Science Trust Asset Allocation Profile Current (Includes Pending distributions and trades) Estimated 11/02/94 Weight Income Yield Income Boney a faLke te eS $ 49.653 $ 2,259 4.55% Principal Mioneya Marketa. 28 5 ee toe ae fe ee $ 83,712 17.64% $ 3,809 4.55% Imcomesnvestments se --— es Sn Ree 2 oe $297,669 62.71% $19,914 6.69% (Growto: Investments ee $ 93,284 19.65% $ 2.316 2.48% $474,665 100.00% $26,038 5.49% Other Principal $ 25,000 [RUST TRS Teh a DT Ue $ 6584 N/A N/A Mineral Interest $ 26,257 N/A N/A Escrow Account $ 57,840 N/A N/A SRG tc | erate ee OTe ee $28,298 4.96% $582,158 60 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) APPENDIX B Report from President-Elect Robert Creek Executive Board Meeting Paducah, Kentucky 3 November 1994 1. Dr. Harry Horner from Iowa State University was invited and accepted our invitation to be the banquet speaker. The title of his talk will be Plants, Fungi and Their “Kidney” Stones. 2. The KERA Symposium will be on Friday at 9:15 and will be moderated by Curtis Wilkins of Western Kentucky University. The speakers will be Dan Ochs, Shirley Wrin- kle, Nelda Freeman, Ben Malphrus and Mike Howard. 3. The Plenary Session was set up by Joe Winstead, who will also moderate the session. The presenters will be Ken Carsten, John MacGregor, Scott Franklin and Wil- liam Bryant. 4. The Agriculture Symposium, Agriculture, Environ- mental Stewardship, and the Role of the Kentucky Acad- emy of Science, will be Thursday at 6:30 P.M. 5. The program has 232 papers being presented with 28 posters. The breakdown of the presentations is listed below: a. Anthropology—15 b. Botany and Microbiology—28 c. Chemistry—17 d. Geography—19 e. Geology—15 f. Physics—10 g. Physiology and Biophysics—16 h. Science Education—13 i. Psychology—22 j- Sociology—5 k. Zoology and Entomology—22 |. Computer Science—6 m. Mathematics—S n. Agricultural Sciences—12 o. Industrial Science—5 p. Cell and Molecular Biology—19 6. I will meet with the sectional officers at 8:00 p.m. Thursday to discuss various topics to include: a. Program b. Abstracts c. Guidelines for student competition d. Duties of sectional officers APPENDIX C Memorandum To: KAS Governing Board From: Larry P. Elliott Subject: President’s Report Date: November 3, 1994 On September 9, 1994, I met with Drs. Bob Creek, Val Dunham, and Curtis Wilkins at Western Kentucky Uni- versity concerning the symposium on KERA for our an- nual Kentucky Academy of Science (KAS) meeting in Pad- ucah. I have met with Dr. Elmer Gray on several occa- sions to facilitate the planning of a symposium entitled “Agriculture: environmental stewardship, and the role of KAS.” From September 29 to October 1, 1994, I attended the 19th Annual Governor’s Conference on the Environment in Louisville, KY. The conference focused on the impor- tance of our forest lands and their interrelationship with the environment, as well as their importance to Kentucky's economic base. While in Louisville, I also attended an- other Kentucky Outlook 2000 meeting on which I serve as a Steering Committee member. The KAS members that were appointed by the Gov- ernor to the Heritage Land Conservation Board were Drs. William S. Bryant and Charles Elliott. All candidates that ran for Vice President, Secretary, and Governing Board Member representatives for the Social Science and Phys- ical Science Division were contacted and cordially thanked for being candidates. The winners were also con- gratulated. They were: Dr. Marcus T. McEllistrem, Vice President, Dr. Peter Z. Armendarez, Secretary, Dr. Jim Hopgood, Social Science, and Dr. Robert Wombles, Phys- ical Science. : All committee chairs were contacted by phone and in writing. Reports that I received will be given under “Standing Committee Reports.” On October 24, 1994, I attended a town meeting at the Regional Conference Center in Bowling Green which was conducted by the Kentucky Long-Term Policy Research Center was created by the General Assembly in 1992 to explore the long-range implications of current policies, emerging issues and trends influencing the Common- wealth’s future. This project is an effort to anticipate 21st Century needs and policy making responses. Hopefully, this total effort will not let Kentucky's future be deter- mined by a select few. TREASURER’S REPORT Kentucky Academy of Science 1994 Starting Balance (January 1, 1994) $91,361.48 ($30,000.00 CD and $61,361.48 divided into Money Market Savings [$58,272.18] and Checking [$3,089.30] Clearance of 1993 Checks. — 98.33 ICO I earn See He + 30,377.30 C.D.’s from Endowment, Athey, and Botany Funds + 81,000.00 +111,278.97 | BG OSU — 20,973.26 Ending Balance (September 30, 1994) $181,667.19 ($181,667.19 divided into Money Market Savings and Checking Account) ACADEMY AFFAIRS 6] Income—1994 Membership Dues —________ $10,144.05 CRUE ied ES $9,794.05 nite. (6) 1994 Se 350.00 Institutional Memberships —__ $12,376.00 Corporate Memberships® 4,845.00 Pees harges 2 690.00 Interest Income _____----____. BankwACCOUMt == ee Interest on Time Deposit — Griffith Memorial Trust 90.51 Wotieell a $30,377.30 ° Includes some Annual Meeting income (registration, banquet fees, Ashland Oil contribution). After Annual Meeting is completed, final report will be submitted. 2,231.74 $1,655.83 575.91 Kentucky Academy of Science Expenses—1994 Rpm $ 2,500.00 INAASS cues== = 90.00 INST 2 ee 250.00 * Awards (Grants—1994) 1,500.00 Georgetown College (Annual Meeting—1993) 2,904.21 (ain a 9 404.95 iransachionse——— =. 7,939.82 Newsletters = es 1,360.63 Anniversary Seals __________ 104.50 Professional Services 583.00 Awoht 2 oes ae 575.00 [DIL fe eae ee 8.00 LI GUNOS Le Se ae 107.30 EGCULIVESeCletaly— = 3,633.80 Postage & P.O. Box Rent — 1,130.24 travel Expense §24.00 iBOG) te ae ee 13.45 Secretarial services 1,011.94 IAT (i em Dees vs es cal 25 654.17 Theta 2 a ee ee $20,973.26 ° Awards were made as follows: $1,000 to Mr. Jeffrey Walck, T. H. Morgan School of Biological Sciences, Uni- versity of Kentucky; $500 to Mr. Randy Mears, Depart- ment of Biological Sciences, Eastern Kentucky Univer- sity. Endowment Fund—1994 Starting Balance (January 1, 1994) _______. $26,784.79 ImCo in Cues omer ey fs DLT Interest dhime WD eposits= a= = 383.94 Bank Account = 127.79 Ending Balance (September 30, 1994) —__ ($20,000.00 CD and $7,296.52 in Money Market § Savings) Botany Fund—1994 Starting Balance (January 1, 1994) $14,229.20 | ha oxOY one See Se ee ee 270.98 Interest (hime Deposits $191.97 Bank Account 79.01 Ending Balance (September 30, 1994) $14,500.18 ($10,000.00 CD and $4,500.18 in Money Market Savings) Marcia Athey Fund—1994 Starting Balance (January 1, 1994) $57,664.59 TC Ome ts See ee eo RR LIU ey interest 222.9. sae ee ime: Deposits =a $979.06 BankeAccoun tae 133.46 Ending Balance (September 30, 1994) —- $58,777.41 ($51,000.00 CD and $7,777.41 in Money Market Savings) Mentor Fund—1994 Starting Balance (January 1, 1994) Int te res ta ee eee ee ) Ending Balance (September 30, 1994) Seoenollevil The above C.D.’s have been cashed and deposted into checking account ready for re-investment in Treasury Notes. $ 3,689.51 Investment in Treasury Notes Two-year notes purchased 10/25/94 $29,000-Rate: 678% Five-year notes purchased 10/25/94 $29.000-Rate: 742% Order placed: 52-week T-Bill $58,000 Rate to be determined 11/10 3-year Treasury Note $29,000 Rate to be determined 11/8 Trans. Ky. Acad. Sci., 56(1—2), 1995, 62-79 HIGHLIGHTS OF THE 1994 ANNUAL MEETING Agriculture, Environmental Stewardship, and the Role of the Kentucky Academy of Science PROGRAM, ANNUAL MEETING KENTUCKY ACADEMY OF SCIENCE 80th ANNUAL MEETING, NOVEMBER 3-5, 1994 Paducah, Kentucky Governing Board Executive Committee President President-Elect Past President Vice President Secretary Treasurer Executive Secretary Editor, Transactions Editor, Newsletter Larry P. Elliott Western Kentucky University Robert O. Creek Eastern Kentucky University Charles N. Boehms Georgetown College William S. Bryant Thomas More College Peter X. Armendarez Brescia College Julia H. Carter Wood Hudson Cancer Research J. G. Rodriguez University of Kentucky Branley A. Branson Eastern Kentucky University Vincent A. DiNoto, Jr. Jefferson Community College Division Representatives and At-Large Members James E. Gotsick (1994) Kimberly W. Anderson (1995) Blaine R. Ferrell (1995) Patricia K. Doolin (1996) David Hogan (1996) Valena Hurt (1996) Gerald L. Demoss (1997) Wimberly C. Royster (1997) J. G. Rodriguez AAAS/NAAS Representative Valgene L. Dunham (1995) Chairperson—K]JAS SYMPOSIUM 6:30 p.mM., 3 November 1994 Mr. Ed Logsdon Topic: How do current agricultural practices in Ken- tucky affect the environment? How could the De- Room—International B Commissioner of Agriculture Morehead State University University of Kentucky Western Kentucky University Ashland Petroleum Company Northern Kentucky University Hazard Community College Morehead State University Kentucky Science & Technology Council, Inc. University of Kentucky Western Kentucky University partment of Agriculture and the Kentucky Acad- emy of Science be mutually supportive? Mr. Phillip Shepherd—Secretary Natural Resources and Environmental Protection Topic: How does the current policy of the Natural Re- sources and Environmental Protection Cabinet affect Kentucky's agriculture? How does your Cabinet view the role of the Kentucky Academy of Science in environmental protection? Mr. Bill Millikan—State Soil Conservationist, USDA Soil Conservation Service PROGRAM, ANNUAL How do SCS policies benefit production agricul- ture and ensure sustained productivity through environmental stewardship in Kentucky? How Topic: could the Kentucky Academy of Science be of assistance to your program? Dr. Luther Hughes—Head, Department of Agriculture, Western Kentucky University Topic: Practically speaking, how does the AG-2000 pro- ject support Kentucky farmers and contribute to environmental stewardship? Does the Kentucky Academy of Science have a role in AG-2000? Dr. Bill Thom—Department of Agronomy, University of Kentucky Topic: How have university researchers responded to needs of Kentucky farmers with respect to en- suring their sustained viability? How has research contributed to a greater lev el of stewardship with respect to the environment? What role do you see for the Kentucky Academy of Science in address- ing these issues? SYMPOSIUM How Will KERA Change Science Education???? 9:15 a.m., 4 November 1994 Room—Kennedy Dr. Dan Ochs—Kentucky Department of Education A brief overview of KERA followed by a discussion of how KERA has changed the approach to science in the high school classroom. \ at Reidland High School, Paducah, KY The impact KERA has had on teaching chemistry and the benefits the students have gained from it. Ms. Nelda Freeman—Biology Teacher at Paducah Tilgh- man High School, Paducah, KY How KERA has changed the teaching of biology and the perceptions that students now have of biology. Dr. Ben Malphrus—Morehead State University How science education is preparing future teachers for KERA. Mr. Mike Howard—Kentucky Science & Technology Council What will be the impact of KERA on college/university faculty and the changes that will be necessary as students enter college after having been education under KERA? The Jackson Purchase Area A Tribute to Raymond H. Athey Kentucky Academy of Science Plenary Session 1:00-2:15 p.m., 4 November 1994 Room—International B Moderator: Joe Winstead—Western Kentucky University “Raymond H. Athey and the Kentucky Academy of Science” MEETING 63 Presenter: Ken Carstens—Murray State University “Archaeological Highlights of Western Kentucky” Presenter: John McGregor—United States Forest Ser- vice “The Fauna of the Jackson Purchase Re- gion: Some Zoogeographic Considerations” Presenter: Scott Franklin—Southern Illinois University “Vegetation History of the Jackson Pur- chase Area Prior to Settlement” Presenter: William S$. Bryant—Thomas More College “Jackson Purchase Vegetation from 1820 to the Present” Speaker for the Annual Awards Banquet The speaker for the Annual Awards Banquet will be Dr. Harry T. Horner, Professor and Director of the Depart- ment of Botany and Bessey Microscopy Facility at lowa State University. The title of his presentation is Plants, Fungi and Their * “Kidney” Stones. In his abstract he states that the biomineralization consists of a series of complex cellular processes by which plants, animals and microor- The more common bio-minerals are the calcium salts, silica ganisms convert solution ions into solid minerals. and iron oxides. Some of the calcium salts and silica serve as major skeletal or shell minerals, whereas others form teeth and serve as ion storage depots for specific physio- logical functions. One calcium salt which is widespread throughout the Plant and Animal Kingdoms that has elud- ed being associated with these and other important struc- tural and physiological functions, is crystalline calcium ox- alate. This mineral can form aggregates or “stones” that are considered pathological in the kidneys and urinary In the higher vascular plants, algae, lichens and fungi, calcium oxalate does not seem to be detrimental. The specialized cells, cellular pro- tracts of animals and humans. cesses, and structures associated with calcium oxalate crys- tal mineralization in all of these latter organisms are unique. Two long-standing ideas are that the crystals may protect the plants against foraging animals and/or humans and, secondly, cry. stal formation ties up the toxic, meta- bolic by-product oxalate formed by the plant. Plant crys- tals have been observed and felt by humans for hundreds and thousands of years and, yet, they are still poorly un- derstood and superficially studied. However, recent stud- ies suggest some interesting possibilities for how this par- ticular fungal and plant biomineralization process may be of benefit to the ecosystem and to humankind, in general. For those that do not attend the banquet but would like to hear Dr. Horner’s presentation, | encourage you to come by after the banquet. There will be chairs available for anyone wishing to hear this interesting presentation. 80th Annual Kentucky Academy of Science Meeting 3-5 November 1994 Paducah Community College Paducah, KY Paducah Community College and Murray State Uni- versity are co-hosting this years meeting. The Executive 64 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) Inn will be the headquarters hotel and all activities will be held there. PROGRAM SUMMARY Thursday, 3 November 1994 1:00 p.m.—4:00 P.M. KAS Governing Board Meeting, Executive Board Room 4:00 p.m.—S:00 P.M. Registration, Main Lobby 6:30 p.m.—S:00 P.M. Symposium—Agriculture, Environmental Stewardship, and the Role of the KAS, International B 7:00 p.m.—8:00 P.M. Meeting of Sectional Officers, Lincoln Room 8:00 p.m.—10:00 P.M. Reception, International D Friday, 4 November 1994 7:30 A.M.—5:00 P.M. Registration, Main Lobby 8:00 a.M.—5:00 P.M. Vendor Exhibits, International D 8:00 a.mM.—5:00 P.M. Scientific Poster Sessions, International D 9:15 a.M.—10:45 a.m. Symposium: How will KERA Change Science Educa- tion???, Kennedy Room 11:00 a.M.—12:30 P.M. Community College Biology Faculty, Jefferson Room 11:00 a.M.—12:30 P.M. Community College Chemistry Faculty, Washington Room 11:00 a.M.—12:30 P.M. Community College Physics Faculty, Truman Room 8:00 a.mM.—9:00 a.M. Sectional Meetings 11:00 a.m.—12:00 noon Sectional Meetings Section B—Botany & Microbiology (8:00), McKinley Room; Section C—Chemistry (11:00), Adams II Room; Section D—Geography (8:00), Roosevelt I Room; Sec- tion E—Geology (8:15), Roosevelt I] Room; Section G—Physiology and Biophysics (11:15), Van Buren Room; Section K—Zoology and Entomology (8:15), Madison Room; Section Q—Agricultural Sciences (8:15), Lincoln Room; Section R—Industrial Sciences (9:25), Eisenhower Room 9:00-9:15 10:45-11:00 Refreshments, International D 12:00-1:00 Lunch, On your own 1:00 p.M.—2:15 P.M. Plenary Session, International B Presiding: Dr. Larry P. Elliott—President, Kentucky Academy of Science Welcome: Dr. Len O’Hara—President, Paducah Com- munity College Announcements: Dr. James Meeks, Chairperson, Local Arrangement Committee Plenary Session: Jackson Purchase Area A Tribute to Ray- mond H. Athey Moderator: Joe Winstead—Western Kentucky University 2:15 p.M.—2:45 P.M. Refreshments, International D 2:45 p.M.—9:00 P.M. Sectional Meetings Section A—Anthropology (2:45), Adams I Room; Sec- tion B—Botany and Microbiology (2:45), McKinley Room; Section C—Chemistry (2:45), Adams II Room; Section D—Geography (2:45), Roosevelt I Room; Sec- tion E—Geology (2:45), Roosevelt II Room; Section G—Physiology and Biophysics (2:45), Van Buren Room; Section H—Science Education (2:45), Washing- ton Room; Section I—Psychology (2:45), Kennedy Room; Section J—Sociology (2:45), Jefferson Room; Section K—Zoology and Entomology (2:45), Madison Room: Section M—Mathematics (2:45), Truman Room; Section Q—Agricultural Sciences (2:45), Lincoln Room 2:30-5:00 p.m. ; Student Reception—sponsored by Murray State Univer- sity Sigma Xi, International B 6:30 P.M.—6:45 P.M. President’s Reception, International B 7:00 p.M.—9:30 P.M. Annual Awards Banquet, International A PLANTS, FUNGI AND THEIR “KIDNEY” STONES, Dr. Harry T. Horner, Professor and Director, Department of Botany and Bessey Microscopy Facility, lowa State University Saturday, 5 November 1994 7:30 a.M.—12:00 noon Registration, Main Lobby 8:00 a.mM.—12:00 noon Scientific Poster Exhibits, International D 8:00 a.m.—12:00 noon Vendor Exhibits, International D 8:00 a.M.—9:30 A.M. Sectional Meetings 11:00 a.m—12:00 noon Sectional Meetings Section A—Anthropology (8:00), Adams I Room; Sec- tion B—Botany and Microbiology (8:00), McKinley Room; Section C—Chemistry (8:00), Adams II Room; Section F—Physics (8:15), Roosevelt I Room; Section G—Physiology and Biophysics (8:15), Van Buren Room; Section H—Science Education (8:00), Washing- ton Room; Section I—Psychology (11:00), Kennedy Room; Section K—Zoology and Entomology (8:00), Madison Room: Section L—Computer Science (8:00), PROGRAM, ANNUAL MEETING 65 Roosevelt If Room; Section M—Mathematies (8:15), Truman Room; Section S (8:00), Eisenhower Room 9:30 a.M.—10:00 A.M. Refreshments, International D 10:00 a.mM.—11:00 a.m. Annual Business Meeting, International B 11:00 a.M.—12:00 noon Sectional Meeting Continued 12:00 noon—1:00 p.m. Lunch, On your own 1:00 p.m.—end Sectional Meetings Cell and Molecular Biology Section I—Psychology, Kennedy Room Section S Room Cell and Molecular Biology, Eisenhower Note: KJAS Each spring the Kentucky Junior Academy of Science holds an Annual Spring Symposium. The 9th Symposium was held at Western Kentucky University in April, 1994. Activities at this meeting include the presentation of Sci- ence Projects by KJAS members as well as Science Bowl competition and Lab Skills competition. The winners of each division of the Science Projects presentations are in- vited to present their work at the annual meeting of the Kentucky Academy of Science. A KJAS precedes the title of each of the papers given by these young scientists. Local Arrangements Committee Paducah Community College Exhibitors Smith & Schaefer, Inc. (Fisher Scientific Co) University of Kentucky Woodson-Tenet Laboratories, Inc. Parco Scientific Co. Ace Glass Incorporated IKA-Works, Inc. Galbraith Laboratories, Inc. Swift Instruments, Inc. Thomas Scientific, Inc. ANTHROPOLOGY James Hopgood—Chairperson Cara Richards—Secretary Room—Adams I Friday, 4 November 1994 James F. Hopgood—Presiding 2:45 P.M. Japan: Impressions, Expectations, and Realities James F. Hopgood—Northern Kentucky University 3:00 P.M. A Site Catchment Analysis of a Prehistoric Site in Wick- liffe, KY April Ke Haneline—Murray State University (Sponsored by Ken Carstens) 3:15 P.M. The Role of Islamic Women in Pondok Pesantrens in East Java Gina Meyer—Northern Kentucky University (Sponsored by James Hopgood) 3:30 P.M. The POW Uprising of 1952: A Participant/Observer’s Per- spective Raymond J. Lewis—Eastern Kentucky University 3:45 P.M. Culinary Syncretism: An Analysis of Yucatecan Bakeries Erin Roberts ards) 4:00 P.M. A Preliminary Analysis of Nail Patterns at Feature 1, 15Lv207, A Civil War Gun Emplacement in Smithland, Kentucky Kathleen Tucker—Murray State University (Sponsored by Ken Carstens) 4:15 P.M. Underutilization of Anthropology in Third World Projects: Centre College (Sponsored by Cara Rich- The Developmental Handicap Julie M. Pelle—Northern Kentucky University (Spon- sored by James Hopgood) 4:30 P.M. The Origins of Australia Dianna Robinson—Murray State University (Sponsored by Ken Carstens) 4:45 P.M. Groundhogs No More: The Joys and Perils of High-Rise Condominium Living James Murray Walker—Eastern Kentucky University Saturday, 5 November 1994 Cara Richards—Presiding 8:00 A.M. Making Kinship: Ceremonial Exchange of Food, Liquor, and Words at Weddings in a Mexican Peasant Village T. D. Murphy—Northern Kentucky University 8:15 A.M. Redistribution and Entrepreneurship in Rural Kentucky: A ‘Big woman's’ Political Strategy Randall Chalk—Northern Kentucky University (Spon- sored by T. D. Murphy) 8:30 A.M. What Function the Saddle-Shaped Mound at the Adams Site in Fulton County, Kentucky? Charles Stout—Murray State University 8:45 A.M. Counterintuitive Findings in a Child Fatalities Study Cara Richards—Transylvania University 9:00 A.M. An Ethnographic Investigation of Antazya’s (Turkey) Street Children: A Preliminary Report 66 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-—2) C. Robert Welch 9:15 A.M. Teaching Anthropology at the Community College: An Applied Approach Tain Barksdale—Madisonville Community College 9:30 A.M. Anthropology Section Business Meeting 9:45 A.M. Refreshments, International D 10:00 A.M. Kentucky Academy of Science Annual Business Meeting, Eastern Kentucky University International B Botany and Microbiology Section Barbara Rafaill—Chairperson Room—MckKinley Friday, 4 November 1994 Barbara Rafaill—Presiding 8:00 A.M. Synopsis of North American Hollies (Aquifoliaceae) Ross C. Clark—Eastern Kentucky University 8:15 A.M. Effects of the Herbicide 2,4-Dichlorophenoxyacetic Acid on Algal Communities in Kentucky Lake: Laboratory and Field Approaches Minoo E. Kobraei and David S$. White—Murray State University 8:30 A.M. Preliminary Report on the Status of the Genus Viburnum in Kentucky Timothy J. Weckman and Ronald L. Jones—Eastern Ken- tucky University 8:45 A.M. Cryptococcus neoformans Serotype Groups Found in Clinical and Environmental Isolates John M.-Clauson and Larry Elliott—Western Kentucky University 9:00 A.M. Refreshments, International D 9:15 A.M. Symposium: How Will KERA Change Science Education, Kennedy Room Moderator: Curtis Wilkins—Western Kentucky University 10:45 A.M. Refreshments, International D 11:00 a.m. Erythronium: The Effect of Mating System on Seed Set, Relationship Between Density and Flowering, and Pol- linator Service Judith Rozeman—Berea College; Tim Weckman and Ron Jones—Eastern Kentucky University 11:15 aM. Preliminary Flora of Shaker Village at Pleasant Hill, Mer- cer County, Kentucky David Taylor—USDA Forest Service 11:30 a.m. Cyperus of Kentucky Randy Mears and Ronald Jones—Eastern Kentucky Uni- versity 11:45 aM. Biomonitoring of Leavenworthia exigua var laciniata (She- pherdsville Glade Cress): A Cedar Glade Endemic Gary Libby, Randy Mears and J. Danny Husband—East- ern Kentucky University 12:00 noon Lunch, On your own 1:00 P.M. Plenary Session: The Jackson Purchase Area—A Tribute to Raymond H. Athey Moderator: Joe Winstead—Western Kentucky University, International B 2:15 P.M. Refreshments, International D Ron Jones—Presiding 2:45 P.M. Morphological Responses of Amur Honeysuckle to Dif- ferent Light Environments Tim Tholemeier—Northern Kentucky University (Spon- sored by James O. Luken) 3:00 P.M. On the Road with Amur Honeysuckle James O. Luken—Northern Kentucky University 3:15 P.M. Micropropagation of American Chestnut Larry A. Giesmann—Northern Kentucky University; Da- vid Bezanson—Jacob Center, Cincinnati Public Schools 3:30 P.M. Effect of Pitcher Size on Fly Symbionts of the New World Pitcher Plants (Sarraceniaceae) Robert Naczi—Northerm Kentucky University 3:45 P.M. Photosynthetic Response to Enhanced Light in Amur Honeysuckle Brian Kunkel—Northern Kentucky University (Sponsored by James Luken) 4:00 P.M. Seedling Responses to Enhanced Light in Amur Honey- suckle Linda Kuddes—Northerm Kentucky University (Spon- sored by James Luken) 4:15 P.M. Analysis of an Old-Growth Upland Wet Woods in Chris- tian County, Kentucky E. W. Chesnut and S. M. Noel—Austin Peay State Uni- versity; J. M. Baskins and C. C. Baskins—University of Kentucky; M. L. McReynolds—Hopkinsville Commu- nity College 4:30 P.M. Kentucky Herbaria: An Update and Outlook Ronald Jones—Eastern Kentucky University PROGRAM, ANNUAL MEETING 67 4:45 P.M. Curator’s Meeting: Discussion of the Future of Kentucky Herbaria and Possible Cooperative Efforts Saturday, 5 November 1994 Barbara Rafaill—Presiding 8:00 A.M. Dogwoods: Molecules, Morphology and Relationships Zack E. Murrell—Western Kentucky University 8:15 A.M. Fossil Diatoms from the Galapagos Islands: Their Use in Reconstructing the History of E] Nino Events Miriam Steinitz Kannan—Northern Kentucky University 8:30 A.M. Protein and Polysaccharide Degradation and Nitrogen Fixation by the Microbial Communities of Pitcher Plants Carrie Gillen—Northern Kentucky University (Sponsored by Miriam Kannan) 8:45 A.M. Geographic Variation in the Purple Pitcher Plant (Sarra- cenia purpurea, Sarraceniaceae ) Brenda Racke—Northern Kentucky University (Spon- sored by Robert Naczi) 9:00 A.M. Trees of Paducah William F. Beasley, Jr—Paducah Community College 9:15 A.M. Phytoplankton Production in a Limestone Quarry in War- ren County, Kentucky Robert C. Molloy and Joe Winstead—Western Kentucky University 9:30 A.M. Refreshments, International D 10:00 a.m. Kentucky Academy of Science Annual Business Meeting, International B 11:00 a.m. RNA Biodiversity: Is Canopy Diversity an Adequate Mea- sure? David D. Taylor—USDA Forest Service; James D. Kis- Eastern Kentucky University 11:15 A.M. Influence of Carbon Source on Growth of Embryogenic Tissue Cultures of Pinus strobus L. Karan Kaul and C. Mahl—Kentucky State University 11:30 a.m. The Isoetes of Kentucky Joe Winstead—Western Kentucky University; Lytton J. Musselman—Old Dominion University 11:45 a.m. KJAS Preventing Ultraviolet Radiation Induced DNA Damage Kate Niehoff—duPont Manual High School (Sponsored by Barbara Fendley) er: 12:00 noon Botany and Microbiology Section Business Meeting CHEMISTRY SECTION Robert Berry—Chairperson Larry Bigham—Secretary Room—Adams II Friday, 4 November 1994 Robert Berry—Presiding 11:00 a.m. Curing Kinetics Study of Epoxy Resins by Thermal Anal- ysis Charles W. M. Lee, Wei-Ping Pan and Jack Li Kentucky University 11:15 A.M. The Effect of the Electric Dipole of Short Peptides on Chemical and Physical Processes Mark Meier and Hemant V. Toshi—University of Ken- tucky 11:30 A.M. The Behavior of Chlorine During Coal Combustion Bryan Travis and Wei-Ping Pan—Western Kentucky Uni- versity 11:45 a.M. Production of Activated Carbon from Used Automobile Tires Susan Hodyclon and Wei-Ping Pan—Western Kentucky Western University 12:00 noon Lunch, On your own 1:00 P.M. Plenary Session: The Jackson Purchase Area to Raymond H. Athey Moderator: Joe Winstead—Western Kentucky University, A Tribute International B 2:15 P.M. Refreshments, International C 2:45 P.M. Reduced-Pressure ICP Detection for Gas Chromatogra- phy M. R. Dunn and H. B. Fannin—Murray State University 3:00 P.M. Prediction of Ash Fusion Temperatures from Elemental Composition Kara Kleeman, Min Guo, W. G. Lloyd and John T. Riley Western Kentucky University 3:15 P.M. Modelling of Excited States Found in the ICP R. S. Perry and J. M. Russell—Murray State University (Sponsored by H. B. Fannin) 3:30 P.M. Design and Characterization of Processable High Tem- perature Thermosetting Polyimide Systems Jenny HeidbrinkK—Western Kentucky University; Gerry 68 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-2) Meyer and James McGrath—Virginia Tech; Wei-Ping Pan—Western Kentucky University 3:45 P.M. ICP-AES Analysis of Solids/Water Slurries Matthew Renfrow, Jody Riley, Bugian Wang and John Riley—Western Kentucky University 4:00 P.M. The Real Route to Solid Deposits in Aircraft Turbine En- gines from Fuel William D. Schulz 4:15 P.M. Using Near Infrared Spectroscopy for Reformer Unit Pro- cess Control Roy R. Bledsoe—Ashland Petroleum Co. (Sponsored by Pat Doolin) 4:30 P.M. College Chemistry be Successful? L. C. Byrd and David R. Hartman—Western Kentucky University Eastern Kentucky University Can We Predict Which Students Will Saturday, 5 November 1994 Larry D. Bigham—Presiding 8:00 a.m. A Route to Transition Metal-Containing Liquid Crystals Jason D. May and Jose M. Workman—Centre College 8:15 A.M. Oxidized Dextran as an Inhibitor of Tooth Decay Timothy E. Davis and Christopher Fletcher—University of Louisville; Karl D. Pryor—du Pont Manual H.S.; Ju- jan Singh, K. G. Taylor and R. J. Doyle—University of Louisville 8:30 A.M. Novel Synthesis of Anti-Cancer Drugs Levi Harper and Gerald Rosenthal—University of Ken- tucky (Sponsored by John Rawls) 8:45 A.M. Modern Technologies Available for the Chemistry Class- room and Laboratory Nedah Rose—John Wiley & Sons, Inc. (Sponsored by Jim Niewahner) 9:00 A.M. KJAS Signal Transduction in Pulmonary Artery Smooth Muscle Cells: The Role of Nitric Oxide Raj Mankad—Paul Lawrence Dunbar (Sponsored by Eliz- abeth Kiknchi) 9:15 A.M. Chemistry Section Business Meeting 9:30 A.M. Refreshments, International D 10:00 a.m. Kentucky Academy of Science Annual Business Meeting, International B GEOGRAPHY SECTION Stuart Foster—Chairperson Wayne Hoffman—Secretary Room—Roosevelt I Friday, 4 November 1994 Stuart Foster—Presiding 8:00 A.M. Sides Cave: The Route to Pike Spring? Christopher Groves—Western Kentucky University 8:15 aM. Central North American Grassland Migration: Stochastic Processes or Climatic Influence? Conrad T. Moore—Western Kentucky University §:30 A.M. Vegetation Stress Analysis Utilizing Remotely Sensed Data—EPA Multi-Media Project—Kentucky Neil Weber—Murray State University 8:45 A.M. The Solstices and Daily Normal Maximum and Minimum Temperature Lags Richard kK. Snow—Western Kentucky University 9:00 A.M. Refreshments, International D 9:15 A.M. Symposium: How Will KERA Change Science Education, Kennedy Room Moderator: Curtis Wilkins—Western Kentucky University 10:45 -A.M. Refreshments, International D 11:00 a.m. Deaths and Injuries from Lightning in Kentucky: Analysis of Location Mary M. Snow—Western Kentucky University 11:15 a.m. Household Filtering and Motive for Household Location in Bowling Green, Kentucky Kendra Vanhooser—Western Kentucky University 11:30 A.M. ; Sinkhole Collapse Subsurface Investigation Using Micro- gravity Nicholas Crawford and Tim Slattery—Western Kentucky University 11:45 a.M. Inner City Revitalization: Bowling Green, Kentucky Kenneth W. McDonald—Western Kentucky University 12:00 noon The History of Kentucky Climate Glen Conner—Western Kentucky University 12:15 p.m. Lunch, On your own 1:00 p.m. Plenary Session: The Jackson Purchase Area to Raymond H. Athey Moderator: Joe Winstead—Western Kentucky University, International B A Tribute PROGRAM, ANNUAL MEETING 69 9:15 P.M. Refreshments, International D Wayne Hoffman—Presiding 2:45 P.M. The Geography of Golf in Kentucky R. L. Marionneaux—Eastern Kentucky University 3:00 P.M. Groundwater Monitoring of Carbonate Aquifers Nicholas C. Crawford—Western Kentucky University 3:15 P.M. Application of GIS in the Power Tool Repair Industry Robin Diederich—Western Kentucky University 3:30 P.M. Transportation Problems and Solutions: An Analysis of Package Transport with United Parcel Service Blake R. Bunner—Western Kentucky University (Spon- sored by James L. Davis) 3:45 P.M. Transportation Analysis of Traffic Accidents in Kentucky in 199] David Murrell—Western Kentucky University (Sponsored by James L. Davis) 4:00 P.M. Retail Stores in Philadelphia: From the City to the Sub- urbs Matthew C. Grecco—Western Kentucky University (Sponsored by James L. Davis) 4:15 p.M. Interregional Migration in Peru Patricia Villalobos—Western Kentucky University 4:30 P.M. Spatial Influences in a Literary Context: A Case Study James L. Davis and Nancy H. Davis—Western Kentucky University 4:45 P.M. Urban Environments, Sense of Place, and the Genesis of Popular Music David J. Keeling—Western Kentucky University 5:00 P.M. The Persistence of Poverty in Kentucky: A Spatial Analysis James M. Bingham and Wayle L. Hoffman—Western Kentucky University GEOLOGY SECTION Deborah Kuehn—Chairperson Kenneth Kuehn—Secretary Room—Roosevelt II Friday, 4 November 1994 Deborah Kuehn—Presiding 8:15 a.m. Computer Aided Instruction in Geology Using Photo- graphs Transferred to CD ROM David R. Dockstader—Jefferson Community College 8:30 A.M. A Conceptual Model for the Occurrence of Hydrochem- ical Facies in an Unmined Area of the Eastern Ken- tucky Coal Field David R. Wunsch—Kentucky Geological Survey 8:45 A.M. Petrology of the Elliott County, Kentucky—Kimberlite In- trusion Brad Maynard—Morehead State University (Sponsored by Robert Boram) 9:00 A.M. Refreshments, International D 9:15 A.M. Symposium: How Will KERA Change Science Education, Kennedy Room Moderator: Curtis Wilkins—Western Kentucky University 10:45 a.m. Refreshments, International D 11:00 a.m. Effect of Fractures on the Availability of Ground Water and the Occurrence of Pesticides and Nitrate in the Epikarst of the Inner Blue Grass Region, Bourbon County, Kentucky Dwayne Keagy and James Dinger—Kentucky Geological Survey; Steven Hampton and Lyle Sendlein—Kentucky Water Resource Research Inst. 11:15 a.m. The Kentucky Ground-Water Data Repository Bart Davidson—Kentucky Geological Survey 11:30 a.M. Characterization and Quantification of Nonpoint-Source Pollutant Loads in a Conduit-Flow-Dominated Karst Aquifer Underlying an Intensive-Use Agricultural Re- gion, Kentucky James Currens—Kentucky Geological Survey 11:15 a.m. Regional Variation of Disconformities in the Mississippian Slade Formation, Northeastern, Kentucky Matthew Vest—Morehead State University (Sponsored by Robert Boram) 12:00 noon Lunch, On your own 1:00 P.M. Plenary Session: The Jackson Purchase Area to Raymond H. Athey Moderator: Joe Winstead—Western Kentucky University, A Tribute International B 2:15 P.M. Refreshments, International D Kenneth Kuehn—Presiding 2:45 P.M. Shear-Related Folds in the Scandinavian Caledonides, Norway Elizabeth A. McClellan—Western Kentucky University 3:00 P.M. Sand and Gravel Resources of the Ohio River Valley Warren Anderson—University of Kentucky (Sponsored by Don Chesnut) 70 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) 3:15 P.M. Implications of a Holocene Mangrove Peat from San Sal- vador, Bahamas Deborah W. Kuehn—Western Kentucky University; Mar- garet Chai—University of Florida 3:30 P.M. Pennsylvanian Sharks from Western Kentucky Coal Area James X. Corgan—Austin Peay State University 3:45 P.M. Sand and Gravel: Ohio River Mile 605 Area, Kentucky— Indiana Graham Hunt—University of Louisville 4:00 P.M. Prospects for Ultra-Clean Kentucky Coals Kenneth W. Kuehn—Western Kentucky University 4:15 P.M. The Geology of Roof Falls in Underground Coal Mines— A Comparison Between Eastern and Western Kentucky David K. Hylbert—Morehead State University 4:30 P.M. Investigation of Biochemical Oxygen Demand Levels on the Campus of the Nicholas County School System and Their Response to Clostridium sporogenes Amanda Abnee—Nicholas County High School (Spon- sored by Kathy Green) 4:45 P.M. Geology Section Business Meeting Puysics SECTION/FALL KAPT MEETING Rico Tyler—Chairperson Vince DiNoto—Secretary Room—Roosevelt I Saturday, 5 November 1994 Rico Tyler—Presiding 9:00 A.M. Extension of I.M.C. Model to Nucleation of Binary Sys- tem H. R. Kobraei and B. R. Anderson—Murray State Uni- versity 9:15 A.M. The Kinematics and HT Structures of Interacting Galax- ies: Implications for the Formation of Blue Dwarf Ir- regulars Benjamin K. Malphrus—Morehead State University 9:30 A.M. A Comparison of the Ultraviolet Spectral Morphology and MK Classifications of B Supergiants in the Small Mag- ellanic Cloud Anthony L. Borchers and Raymond C, McNeil—Northern Kentucky University 9:45 A.M. A Remarkable Comet: Shoemaker-Levy 9 Roger Scott, Richard Hackney, Karen Hackney and Jim Parvin—Western Kentucky University 10:00 a.m. Kentucky Academy of Science Annual Business Meeting, International B 11:00 a.m. The Kentucky Space Grant Consortium Program into the Year 2000 Karen Hackney, Richard Hackney, Roger Scott and Tom Bohuski—Westem Kentucky University 11:15 a.m. OHMIC Contacts to SiC John Crofton—Murray State University 11:30 a.m. Physics Section/KAPT Business Meeting Reports: President—Rico Tyler; Treasurer—Vince Di- Noto; Survey—Chris Graney; Other reports 11:45 A.M. Effects of lon Damage on the Superconductors Current- Voltage Relationships in Strong Magnetic Fields in NbSe,,. J. G. Childers and W. F. Henshaw—Union College; Jungq- ing Zhang and L. E. DeLong—University of Kentucky; R. C. Budhani—Brookhaven National Laboratory 12:00 noon : Analysis of Laser Induced Fluorescence Data from Rock- et Motor Exhausts Stephen M. Fuqua and Stephen H. Cobb—Murray State University 12:15 P.M. Post-Impact Vehicle Lamp Filament Examination as an Accident Reconstruction Technique Alan A. Johnson and David Harmeling—University of Louisville 12:30 P.M. KJAS Photoelectrochemical Cells Sallie Rademacher—Notre Dame Academy (Sponsored by Sisters Averbeck and Parrott) PuystoLocy, BiopHysics, BIOCHEMISTRY, AND PHARMACOLOGY SECTION Suzanne Byrd—Chairperson Chang Wang—Secretary Room—Van Buren Friday, 4 November 1994 Suzanne Byrd—Presider 11:15 A.M. Mathematical Modeling of Primary Production in Ken- tucky Lake B. R. Anderson, H. R. Kobraei and Mary C. Child—Mur- ray State University 11:30 a.m. Glycoconjugate Expression in Normal and Regenerating Lateral Line Sensory Epithelia of the Axolotl Salaman- der, Ambystoma mexicanum PROGRAM, ANNUAL MEETING ial Richmond ]. Miller and Kk. J. Balak—Western Kentucky University 11:45 a.m. Experimental Approaches to Investigate Neuromast Re- generation of the Axolotl Lateral Line R. R. Speck and K. J. Balak—Western Kentucky Univer- sity 12:00 noon Lunch, On your own 1:00 P.M. Plenary Session: The Jackson Purchase Area—A Tribute to Raymond H. Athey Moderator: Joe Winstead—Western Kentucky University, International B 2:15 P.M. Refreshments, International D 2:45 P.M. The Steady-State Kinetics of Lithium (Li+) Inhibition of Yeast Enolase William W. Farrar 3:00 P.M. Taste Comparison Between Smoked Paddlefish and Cat- fish Meat Changzheng Wang and Steve D. Mims—Kentucky State University 3:15 P.M. Transcutaneous Collection and Quantitative Measurement of Biological Hazards Michael Timmons—Western Kentucky University; Stev- Eastern Kentucky University en Collier and Sorell Schwartz—Georgetown Uni- versity; Claire Rinehart—Western Kentucky Univer- sity 3:30 P.M. Isozymes of Enolase in Pig Tissue Vince Barrows and William Farrar—Eastern Kentucky University 3:45 P.M. Consumer Factors Related to the Frequency of Fish Con- sumption Changzheng Wang and Steve D. Mims—Kentucky State University 4:00 P.M. Purification and Mechanism Studies of Pyruvate Kinase from the Flight Muscle of the House Sparrow (Passer domesticus) William W. Farrar—Eastern Kentucky University 4:15 P.M. Effect of Protein and Bicarbonate Intake on Kidney Min- eral Contents of Fisher 344 Rats Changzheng Wang, C. J. Lee and Abbas Babalmoradi— Kentucky State University 4:30 P.M. Steady-State Mechanism of Lactate Dehydrogenase from House Sparrow Flight Muscle and Heart William Farrar and Young Jo Farrar—Eastern Kentucky University 4:45 P.M. Effects of Nutrients and Retention Time on the Primary Production of Kentucky Lake Mary C. Child, H. R. Kobraei and B. R. Anderson—Mur- ray State University Saturday, 5 November 1994 Chang Wang—Presider 8:15 A.M. Apoptosis in Differentiated and Undifferentiated Murine F9 Teratocarcinoma Cells E. Deaton, S. Vogelpohl, B. Baldwin, R. Snyder and J. Carter—Wood Hudson Cancer Research Laboratory 8:30 A.M. The Role of Arginine in Pinene Synthases of Salvia offi- cinalis Steve Hume—Centre College; Thomas Savage and Rod- ney Croteau—Washington State University; Linda Rob- erts—Centre College 8:45 A.M. Sclerotinia Shows Mating Type Homology to Both Het- erothallic and Homothallic Fungi Martin Brock—Kastern Kentucky University 9:00 A.M. Investigation of the Effect of Methionine Oxidation on the Conformation of Soluble and Lipid-Bound Apolipopro- tein A-1 by Limited Proteolysis Michelle Reader—Centre College; Linda Roberts—Whit- man College; Christie Brouillette—Southerm Research Institute 9:15 A.M. Physiology, Biophysics, Biochemistry, and Pharmacology Section Business Meeting 9:30 A.M. Refreshments, International D 10:00 A.M. Kentucky Academy of Science Annual Business Meeting, International B SCIENCE EDUCATION SECTION Robert Boram Peter V. Lindeman—Secretary Chairperson Room—Washington Friday, 4 November 1994 Robert Boram—Presiding 2:45 P.M. Are Kentucky Students Sleeping Enough? John G. Shiber—Prestonsburg Community College 3:00 P.M. KERA, “Fast Plants,” and Curriculum Integration: What do These Things Have in Common? Kim Alexander and Robert Creek University 3:15 P.M. The Gender Equitable Classroom—Instruction That Im- proves Learning for Male and Female Students Eastern Kentucky M2, TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) Robert Boram and Joyce Saxon—Morehead State Univer- sity 3:30 P.M. Conservation Biology for the College Freshman Non-Sci- ence Major Peter V. Lindeman—Madisonville Community College 3:45 P.M. Development of Performance Assessment Tasks for Fu- ture Science Teachers Vince DiNoto—Jefferson Community College; Valgene Dunham—Westerm Kentucky University 4:00 P.M. Scientists of Kentucky: An Oral History Project Howard Powell and Morris Taylor—Eastern Kentucky University 4:15 p.m. A Pilot Project Pairing an Introduction to College Science with Intermediate Algebra David Dockstader and Brita Dockstader—Jefferson Com- munity College 4:30 P.M. Effect of Collaborative Activities on Grade Distribution in Biology Veena Sallan—Owensboro Community College Saturday, 5 November 1994 Peter V. Lindeman—Presiding 8:00 A.M. Penny Experiments for General Chemistry Harry Conley—Murray State University; James Meeks— Paducah Community College 8:15 aM. A Model Classroom Curriculum Integrating the Core Concepts of Biomedical Research and Societal Values to Achieve KERA Outcomes in the Secondary Science Classroom Stephanie Wyatt and Cloyd Bumgardner—Calloway County High School; Gregory Popken—University of North Carolina 8:30 A.M. Use of Gel Permeation Chromatography in the Under- graduate Polymer Laboratory Jon Doyle, Kevin Petal and Thomas Green—Western Kentucky University 8:45 A.M. Polymer Synthesis in the Undergraduate Polymer Labo- ratory Kevin Petal, Jon Doyle and Thomas Green—Western Kentucky University 9:00 A.M. Enhancing Student Perceptions of Science and Achiev- ing Kentucky Education Reform Act Learner Out- comes Through a Creative Science Classroom Curric- ulum Stephanie Wyatt and Cloyd Bumgardner—Calloway County High School 9:15 A.M. Science Education Section Business Meeting 9:30 A.M. Refreshments, International D 10:00 A.M. Kentucky Academy of Science Annual Business Meeting, International B PsyCHOLOGY SECTION Terry Barrett—Chairperson Jeff Smith—Secretary Room—Kennedy Friday, 4 November 1994 Terry Barrett—Presiding 2:45 P.M. Gender Differences in Public Speaking Anxiety Tina Lane—Murray State University (Sponsored by Terry Barrett) 3:00 P.M. Longlasting Behavioral and Neurochemical Consequences of Chronic Apomorphine Treatments Tracey Ellison—Morehead State University; Patricia Ro- binet—University of Kentucky; Bruce Mattingly— Morehead State University; Mike Bardo—University of Kentucky 3:15 P.M. Reality Characteristics of Hallucinations Reported by Normal Adults and by Chronic Schizophrenics Terry R. Barrett—Murray State University 3:30 P.M. The Personality Characteristics Associated with Alienation in College Students Valerie Haire—Murray State University (Sponsored by Terry Barrett) 3:45 P.M. Mental Practices: An Undergraduate Sample Danny R. Adamson—Murray State University (Sponsored by Terry R. Barrett) 4:00 P.M. A Timely Look at the Fundamentals of Academic Pro- crastination Christina E. Smith—Murray State University (Sponsored by Terry R. Barrett) 4:15 P.M. Effects of Coaching Style on Competition Performance in Children Beverly Lenicky—Northern Kentucky University (Spon- sored by David Hogan) 4:30 P.M. Emotion Affects Memory Kathy Alexander—Murray State University (Sponsored by Terry R. Barrett) 4:45 P.M. Women’s Perceptions of Female Gender Roles PROGRAM, ANNUAL MEETING (8) K. Eleanor Sheets—Murray State University (Sponsored by Terry R. Barrett) 5:00 P.M. Effects of Response Prevention on the Development of Behavioral Sensitization to Apomorphine Sonia Fields, Steve McDonald and Bruce Mattingly— Morehead State University Saturday, 5 November 1994 Jeff Smith—Presiding 11:00 a.m. Effects of Chronic Dopamine D3 Receptor Stimulation of Locomotor Activity and Subsequent Sensitivity to Ap- omorphine Mike Cecil, Sonia Fields, Steve McDonald and Bruce Mattingly—Morehead State University 11:15 A.M. Time Urgency Between Type A and B Coronary Prone Behavior Michael S$. Dykes—Murray State University (Sponsored by Terry R. Barrett) 11:30 A.M. The Effects of a Newsletter on Increasing Familiarity Among Fast Food Restaurant Employees to Improve Teamwork Kara J. Collins—Northern Kentucky University (Spon- sored by David Hogan) 11:45 a.m. Neurochemical and Behavioral Consequences of Chronic Dopamine D3 Receptor Stimulation Mike Langfels—Morehead State University; Patricia Ro- binet—University of Kentucky; Bruce Mattingly— Morehead State University; Mike Bardo—University of Kentucky 12:00 noon Lunch, On your own 1:00 P.M. The Relationship Between High Versus Low Imagery Re- call and Performance in an Introductory Psychology Course Francis H. Osborne and Aimee C. Lawson—Morehead State University 1:15 p.m. Alcohol Expectancies of College Students Joe D. Littleton—Murray State University (Sponsored by Terry R. Barrett) 1:30 P.M. . A Cross Cultural Comparison of Posture Perception Kim Kelly and Jack Thompson—Centre College 1:45 P.M. Personality Correlates of Performance in an Introductory Psychology Course Francis H. Osborne and Steven E. Dolan—Morehead State University 2:00 P.M. Sex and Age Effects on Grouping Behavior and Interper- sonal Touch Kim Kelly, Phyllis Passariello and Jack Thompson—Cen- tre College 2:15 P.M. The Effect of Anxiety on Dysmenorrhea in College Stu- dents Valeria S. Wagoner and Frank H. Osborne—Morehead State University 2:30 P.M. Self-Esteem of Black, White, Male, and Female College Students Laura Engle and Jack Thompson 2:45 P.M. KJAS Transformation of a Personality: A Three Year Study Centre College Phonesavane Liankehammy—Warren Central High School (Sponsored by Linda Walker) 2:45 P.M. Psychology Section Business Meeting SOCIOLOGY SECTION J. Allen Singleton—Chairperson Steve Savage—Secretary Room—Jefferson Friday, 4 November 1994 J. Allen Singleton—Presiding 2:45 P.M. The Electoral College: History and Prospects Tamatha Brewer—Eastern Kentucky University (spon- sored by J. Allen Singleton) 3:00 P.M. Environmental Consciousness and Awareness: A Quanti- tative Analysis of University Students Molly B. Kerby—Western Kentucky University (spon- sored by Joe Winstead) 3:15 P.M. Single Male Parenting Kelly Davis—Eastern Kentucky University (sponsored by Steve Savage) 3:30 P.M. Beyond the Headlines: Examining Ethics and the Ken- tucky General Assembly Amy Etmans—Eastern Kentucky University (sponsored by J. Allen Singleton) 3:45 P.M. Urban-Rural Aspects of Kentucky Politics J. Allen Singleton—Eastern Kentucky University 4:00 P.M. Sociology Section Business Meeting ZOOLOGY AND ENTOMOLOGY SECTION Guenter Schuster—Chairperson Gordon Weddle—Secretary Room—Madison 74 TRANS. KENTUCKY ACADEMY OF SCIENCE 56( 1-2) Friday, 4 November 1994 Guenter Schuster—Presiding 8:15 A.M. Daily Pattern of Brain GABA Levels in the Cockroach, Leucophaea maderae Juli McCay, Kimberly Romero, Blaine Ferrell and Darwin Dahl—Western Kentucky University 8:30 A.M. The Freshwater Unionids (Mussels) of Marsh Creek, McCreary County, Kentucky Ronald R. Cicerello and Ellis Laudermilk—Kentucky Na- ture Preserve Commission 8:45 A.M. Coccinelidae Predation of First and Second Generation European Corn Borer, Ostrinia nubilalis, Egg Masses Kentucky Field Corn Donna Shanklin, Douglas Johnson, Lee Townsend, Jr. and Ric Bessin—University of Kentucky 9:00 A.M. Refreshments, International D 9:15 A.M. Symposium: How Will KERA Change Science Education, Kennedy Room Moderator: Curtis Wilkins—Western Kentucky University 10:45 aM. Refreshments, International D 11:00 a.m. Changes in Gut Capacity due to Increased Energy De- mand in the Pine Vole (Microtus pinetorum) Mitzi Austin and Terry Derting—Murray State University 11:15 a.m. Development of Sawtoothed Grain Beetle and Red Flour Beetle on Corn Previously Infested by Angoumous Grain Moth Patti Rattlingourd and Paul Weston—Kentucky State Uni- versity 11:30 a.m. A Comparison of the Seasonal Variation of Intestinal Villi Morphology of a Herbivorous Species (Microtus penn- sylvanicus) and an Omnivorous Species (Peromyscus leucopus) Arthur Scott and Terry Derting—Murray State Univer- sity 11:45 a.m. Potential Mode of Action on Host Development of a Se- cretory Product from a Parasite D. L. Dahlman, E. J. Schepers, Z. Zhang and F. A. Di- Luna—University of Kentucky 12:00 noon Lunch, On your own 1:00 P.M. Plenary Session: The Jackson Purchase Area—A Tribute to Raymond H. Athey Moderator: Joe Winstead—Western Kentucky University, International B 2:15 P.M. Refreshments, International D 2:45 P.M. Similar Seasonal Changes in Gut Morphology in a Her- bivore (Microtus pennsylvanicus) and a Granivore (Peromyscus leucopus) Edward Noakes and Terry Derting—Murray State Uni- versity 3:00 P.M. Exciting Discoveries in the Lepidoptera Fauna of the Jackson Purchase, Kentucky: A Review Charles Covell, Jr and William Black, Jr—University of Louisville 3:15 P.M. The Impact of Red-Cockaded Woodpecker Forest Man- agement Practices on Populations of White-Footed Mice (Peromyscus leucopus) Raymond D. Campbell—University of Kentucky; David Nusbaumer—Elizabethtown Community College 3:30 P.M. Food Habits and Distribution of Larval Bluegill, Lepomis macrochirus, in Ledbetter Embayment, Kentucky Lake M. J. Brouder and T. J. Timmons—Murray State University 3:45 P.M. Density and Relative Abundance of the River Cooter (Pseudemys concinna) in Kentucky Lake and tlhe Lower Tennessee River: A Spotting-Scope Survey Peter V. Lindeman—Madisonville Community College 4:00 P.M. Equine Protozoal Myeloencephalitis: A Review Shelby Stamper and D. E. Granstrom—University of Kentucky 4:15 P.M. Conservation Status and Systematics of the Palezone Shin- er, Notropis albizonatus (Pisces: Cyprinidae), a Feder- ally Endangered Species in Kentucky Brooks Burr and Kenneth Cook—Southern Illinois Uni- versity at Carbondale; Melvin Warren—Southern For- est Experiment Station, Oxford, Mississippi 4:30 P.M. Distribution, Reproduction and Recovery of the Relict Darter, Etheostoma chienense (Pereidae), in Western Kentucky 4:45 P.M. Intraspecific Phylogeography of Five North American Highland Fishes: A Test of the Pleistocene Vicariance Hypothesis Rex Meade Strange and Brooks M. Burr—Southern Illi- nois University; Kyle Piller and Brooks M. Burr— Southern Illinois University at Carbondale Saturday, 5 November 1994 Gordon Weddle—Presiding 8:00 A.M. Microhabitat Preferences of Larval Studfishes (Fundulus catenatus): Impact of Predator Presence PROGRAM, ANNUAL MEETING Ths: Will Sudduth and Mike Barton 8:15 A.M. The Identification, Distribution, and Habitat of Anguis- pira rugoderma Hubricht, A Land Snail Endemic to Southeastern Kentucky John MacGregor—USDA Forest Service; James Kiser— Eastern Kentucky University 8:30 A.M. The Influence of Salinity and pH on the Rate of Heart Beat and Locomotor Activity of Daphnia magna Jeff Reynolds—Moss Middle School; Robert Hoyt—West- em Kentucky University 8:45 A.M. The Bluegrass and Western Coal Fields: Filter Barriers to the Dispersal of Kentucky Vertebrates Centre College Les Meade—Morehead State University; John Mac- Gregor—USDA Forest Service 9:00 A.M. Factors Affecting Gas Exchange in Gila Woodpecker Nest-Cavities David Nusbaumer—Elizabethtown Community College 9:15 A.M. KJAS Effectiveness of Alternative Insecticide Formulations on the German Cockroach, Blatena germanica Burr Settles—Lafayette High School (Sponsored by James Gentry) 9:30 A.M. Zoology and Entomology Section Business Meeting 9:45 A.M. Refreshments, International D 10:00 A.M. Kentucky Academy of Science Annual Business Meeting, International B COMPUTER SCIENCE SECTION Sylvia C. Pulliam—Chairperson Richard A. Rink—Secretary Room—Roosevelt II Saturday, 5 November 1994 Sylvia Pulliam—Presiding 8:00 A.M. Fuzzy Logic Controller Projects for Students Art Shindhelm—Western Kentucky University 8:15 A.M. Data Compression: Theory and Practice John Crenshaw—Western Kentucky University 8:30 A.M. Using Algorithm Animation in CS1 and CS2 Carol W. Wilson—Western Kentucky University 8:45 A.M. Teaching Computer Science in Russia Sylvia Clark Pulliam—Western Kentucky University 9:00 A.M. Open Labs for CSC Using Graphical Displays of Algo- rithms for Large Integer Arithmetic Kenny Napier and Bill Janeway versity 9:15 A.M. Cards/Permutations: Open Labs and Analysis of Algo- rithms Eastern Kentucky Uni- Bill Janeway and Donnie Grimes—Kastern Kentucky University 9:30 A.M. Refreshments, International D 10:00 A.M. Kentucky Academy of Science Annual Business Meet- ing MATHEMATICS SECTION Carroll Wells—Chairperson Russell Brengelman—Secretary Room—Truman Friday, 4 November 1994 Carroll Wells—Presiding 2:45 P.M. Common Threads of Math Reform Movements Joseph F. Stokes—Western Kentucky University 3:00 P.M. Using “Derive” in Calculus Kathy Kepner—Paducah Community College; Maura Corley—Henderson Community College 3:15 P.M. “Hands on” Geometry Activities Carroll G. Wells—Western Kentucky University 3:30 P.M. Using the Genetic Algorithms James Porter—Western Kentucky University Saturday, 5 November 1994 Russell Brengelman—Presiding 8:15 A.M. A Pilot Project Pairing Intermediate Algebra with an In- troductory Science Course Brita Dockstader—Jefferson Community College 8:30 A.M. Parts from Dirichlet J. B. Barksdale, Jr—Western Kentucky University 8:45 A.M. Developing Actuarial Science and Operations/Quality Control Curricula in Applied Mathematics Alan D. Smith—Robert Morris College 9:00 A.M. An Examination of a Partial Differential Equation from Underwater Acoustics Mark P. Robinson—Western Kentucky University 9:15 A.M. Mathematics Section Business Meeting 9:30 A.M. Refreshments, International D 76 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) 10:00 a.m. Kentucky Academy of Science Annual Business Meeting, International B AGRICULTURAL SCIENCES SECTION Matthew E. Byers—Chairperson Elmer Gray—Secretary Room—Lincoln Friday, 4 November 1994 Matthew E. Byers—Presiding 8:15 A.M. Ecology of Insects in Two-Year-Old On-Farm Stored Shelled Corn J. D. Sedlacek, P. A. Weston, B. D. Price and P. L. Rat- tlingourd—Kentucky State University 8:30 A.M. Effects of Soybean Cultivars and Planting Dates on Bio- mass Production Aslam Tawhid and Elmer Gray—Western Kentucky Uni- versity 8:45 A.M. Effects of Winter-Cropped Hairy Vetch and Tillage on Ni- trogen Nutrition of Sweet Corn and Watermelon Anthony Silvernail and Gary Cline—Kentucky State Uni- versity 9:00 A.M. Refreshments, International D 9:15 A.M. Symposium: How Will KERA Change Science Education, Kennedy Room Moderator: Curtis Wilkins—Western Kentucky University 10:45 A.M. Refreshments, International D 11:00 a.m. A Standardized Comparison of Population Characteristics of Freshwater Prawns, (Macrobrachtium rosenbergii), Raised in Two Different Geographical Regions (Ken- tucky and Mississippi) James Tidwell and Shawn Coyle—Kentucky State Uni- versity; Louis R. D’Abramo— Mississippi State Univer- sity 11:15 a.m. Dacthal Persistence in Soil Nekiya Baker, Matt Byers, George Antonious and Debra Hilborn—Kentucky State University 11:30 a.M. Analysis of Esfenvalerate on Vegetables Dawn Greene, George Antonious and Matthew E. Byers—Kentucky State University 11:45 a.m. Vegetable Culture on Erodible Land Debra Hilborn, Matthew Byers and George Antonious— Kentucky State University 12:00 noon Lunch, On your own 1:00 a.m. Plenary Session: The Jackson Purchase Area—A Tribute to Raymond H. Athey Moderator: Joe Winstead—Western Kentucky University, International B 2:15 P.M. Refreshments, International D Elmer Gray—Presiding 2:45 P.M. Sustainable Agronomic Research Projects at Western Kentucky University Elmer Gray, Aslam Tawhid, Tena Wright and Brian Lace- field—Western Kentucky University 3:00 P.M. The Influence of Incident Light Intensity and Fertilization on Growth and Development of Pawpaw (Asimina tri- loba Dunal) Seedlings Lakeasha Jones, Desmond Layne and Michael Kwantes— Kentucky State University 3:15 P.M. Influence of Mulch Color on Yield of Sweet Basil E. Greer, K. Kaul and C. L. Mahl—Kentucky State Uni- versity 3:30 P.M. Environmental Effects on Indian Mock Strawbetry Brian D. Lacefield and Elmer Gray—Western Kentucky University 3:45 P.M. The Street Tree Inventory—Putting the Data to Work James M. Martin—Westerm Kentucky University 4:00 P.M. Agricultural Sciences Section Business Meeting INDUSTRIAL SCIENCE SECTION Burton H. Davis/Patricia K. Doolin— Co-Chairpersons Room—Eisenhower Friday, 4 November 1994 9:25 A.M. Introduction 9:30 A.M. Biological Research in the Electric Utility Industry: The Search for an Answer to the Electromagnetic Field Di- lemma Jeff West—East Kentucky Power Corporation 10:00 a.m. Method to Reduce Diesel Engine Exhaust Emissions Greg Garr—I.C.T. Incorporated 10:30 A.M. Petroleum Products in Your Everyday Life Robert Wombles—Ashland Petroleum Company 11:00 a.m. Ground Water Investigation and Clean Up Colleen Winker, P.E.—Martin Marietta PROGRAM, ANNUAL MEETING al 11:30 a.m: Trans-Isomers Minimization During the Catalytic Hydro- genation of Vegetable Oils John H. Hasman and Patrick McLaughlin—United Cata- lysts Inc. 12:00 noon Closing Remarks CELL AND MOLECULAR BIOLOGY John Rawls—Chairperson Claire Rinehart—Secretary Room—HEisenhower Saturday, 5 November 1994 John Rawls—Presiding 8:00 A.M. A Cytokinin-Responsive mRNA in Phaseolus vulgaris L. Kerrie L. McDaniel—Westem Kentucky University; Da- vid Lightfoot—Southern Illinois University 8:15 a.M. Molecular Dissection of the Mouse Alpha-Fetoprotein Repressor Region Cassandra Backer, Amy Ellis and Brett Spear (Sponsored by John Rawls)—University of Kentucky 8:30 A.M. Mapping the Location of Somatostatin-Like Immunoreac- tive Cells in Tadpole Tectum Angie Baker and Elizabeth Debski (Sponsored by John Rawls)—University of Kentucky 8:45 A.M. Studies of Non-Protein Amino Acid Disruption in Pro- teins Michael Bass and Gerald Rosenthal (Sponsored by John Rawls)—University of Kentucky 9:00 A.M. Intracellular Niacin Status and Mechanisms of Carcino- genesis Christopher Watt and Elaine Jacobson (Sponsored by John Rawls)—University of Kentucky 9:15 a.M. Influence of Antioxidant and Temperature Treatment on Lipid Peroxidation and Senescence in Broccoli Florets Sau-Min Tam, Hong Zhuang and Margret Barth (Spon- sored by John Rawls)—University of Kentucky 9:30 A.M. Refreshments, International D 10:00 a.m. Kentucky Academy of Science Annual Business Meeting, International B 11:00 a.m. Studies of the Variation of a Surface Protein of Strepto- coccus equi Subspecies zooepidemicus Matt Blair, John Walker and John Timoney (Sponsored by John Rawls)—University of Kentucky 11:15 a.m. Investigation of Repressor Dependent Segregation in Escherichia coli Michael Zgoda and Don Bick (sponsored by John Rawls)—University of Kentucky 11:30 A.M. Modulation of TNF Cytotoxicity by NAD. Mandy Wilson, David Hiestand, Boyd Haley, Elaine Ja- cobson and John Rawls—University of Kentucky 11:45 a.M. The Influence of Methyl Jasmonate on Lipid Peroxidation and Deterioration in Broccoli D. Gueorguieva, H. Zhuang and M. Barth (Sponsored by John Rawls)—University of Kentucky 12:00 noon Lunch, On your own Claire R. Rinehart—Presiding 1:00 p.m. Roles of Polyunsaturated Fatty Acids in Neuronal Death: Lipid Peroxidation and Arachidonic Acid Metabolism Irene Hong and Marion Steiner (Sponsored by John Rawls)—University of Kentucky 1:15 P.M. Effect of Ozone Exposure on Antioxidant Vitamins in Fruit Tissue Kellie Kute, M. Margaret Barth and Zhou Chen (Spon- sored by John Rawls)—University of Kentucky 1:30 P.M. Regulation of Expression of Protease Nexin 1 in Neural Tissue: Relationship to Alzheimer’s Disease Paul Jett, John Rawls and Stephen Zimmer—University of Kentucky 1:45 P.M. Interactions of the Tobacco Vein Mottling Virus’ Cylin- drical Inclusion Protein with Other Virus Proteins Beth Smith and Arthur Hunt (Sponsored by John Rawls)—University of Kentucky 2:00 P.M. The Organelle Specific Role of the Delta-9 Fatty Acid Desaturase Michael Scowby, Sergei Avdiushko and David Hildebrand (Sponsored by John Rawls)—University of Kentucky 2:15 P.M. Fatty Acid Effects on Vitamin Levels in Cultured Endo- thelial Cells Brad Middendorf and Bernhard Hemig (Sponsored by John Rawls)—University of Kentucky 2:30 P.M. The Effects of Teratocyte Secretory Products on Protein Synthesis by Fat Body in Various Larval Stages of He- liothis virescens Lisa McGraw and Douglas L. Dahlman Kentucky 2:45 P.M. How Do Lymphocytes Recognize Antigens in Immune Privileged Sites? Loh, Wai Khan (Sponsored by John Rawls)—University of Kentucky University of 78 TRANS. KENTUCKY ACADEMY OF SCIENCE 56( 1-2) 3:00 P.M. The Synthesis of Insulin-Like Growth Factor Binding Pro- tein-2 mRNA in Human T-Cells Sandhya Venugopal, Lorri Ann Morford and Thomas Roszman (Sponsored by John Rawls)—University of Kentucky 3:15 P.M. Cell and Molecular Biology Section Business Meeting SECTIONAL POSTERS Posters will be available for viewing for the duration of the meeting. Presenters are requested to be at their post- ers to facilitate discussion of their research at the follow- ing times: Friday, November 4 from 2:15 P.M. to 3:00 P.M.; Saturday, November 5 from 9:00 a.m. to 10:00 a.m.; Sat- urday, November 5 from 11:00 a.m. to 12:00 noon. il ~ 10. The Effect of Agricultural Chemicals on Ground-Wa- ter Quality in Lacustrine Deposits of the Western Kentucky Coal Field Philip Conrad and James Dinger—Kentucky Geolog- ical Survey; Lyle Sendlein—Water Resources Re- search Institute . A Study on Dental Health Professionals Who Provide Care to the Homeless Arthur Van Steward and Eric Veal—University of Louisville Dental School . Decomposition of Nerve Gas and Mustard Gas Ana- logs Using Nicotine and Ultrasound John Meisenheimer, Lawrence Miller and William Schulz—Eastern Kentucky University . Influence of Constructed Wetlands on Pathogen and Nutrient Removal Frank Young II, Matt Byers, George Antonious and Arla Burks—Kentucky State University . Elkhom Creek Monitoring Program: Influence of Livestock on Water Quality Arla Burks, Frank Young, Dan Logan and Matt Byers—Kentucky State University . Biological Control Using Interplantings Patty Lucas and Doug Johnson—University of Ken- tucky . Fullerenes and Fullerene Derivatives: Preparative Chromatography of Higher Fullerenes and Synthesis of Fullerene Derivatives Rebecca Massie and B. Henshaw—Union College; J. Selegue—University of Kentucky . Effects of Acidified Field Soil on Sericea Lespedeza Inoculated with Bradyrhizobium G. R. Cline and A. F. Silvernail—Kentucky State Uni- versity . A Theoretical Study of Molecular Geometries and Harmonic Vibrational Frequencies: Ge2C and GeC2 Regina Rao—Gannon University; Roger Grey (Men- tor)—University of Kentucky (Sponsored by Sylvia Daunert) Comparisons of Composts for Growing Vegetables in Containers 11. 13. 14. 15. 16. live 18. 19. Aslam Tawhid and Elmer Gray—Westermm Kentucky University Container Herb Production Using Compost Mixtures Tena Wright and Elmer Gray—Western Kentucky University . Treating Livestock Waste Waters with Constructed Wetlands D. A. Stiles, B. Kessler, O. W. Dotson II, B. Basham, R. Johnson and A. Bedel—Western Kentucky Uni- versity NMR Spectra of Sulfonium Salts in Presence of Lan- thanide Shift Reagents Bijan Radmard, Thomas Green and Lester Pester- field—Western Kentucky University Plant (Wheat) and Soil Analyses Following High Rates of Manure Application on Corn R. Johnson, B. Basham, B. Kessler, O. W. Dotson III, D. Stiles and A. Bedel—Westem Kentucky University Mechanistic Investigation of Imine Radical Anions David Eaton, Robert Hendricks, Brian Grace, Ken Meridith and R. W. Holman—Western Kentucky University Habitat pH Preference of Salamanders in Madison County, Kentucky Cara Lin Bridgman—Eastern Kentucky University (Sponsored by Paul Cupp) : Responses of the Ground Skink, Scincella lateralis, to Chemical Deposits of Snakes Elizabeth R. Gosnell and Paul Cupp, Jr.—Eastern Kentucky University A Mechanistic Investigation of Gas-Phase Heteroar- omatic Electrophilic Aromatic Substitutes Ed Whittle and R. W. Holman—Western Kentucky University Salamander Responses to Chemical Deposits of Pred- atory and Nonpredatory Snakes Paul V. Cupp, Jr—Eastern Kentucky University . Antisickling Properties of Phenylalanine and Hydrox- ybenzoic Acid Extracts of Lima Beans (Phaseolus lun- atus) Felix Akojie—Paducah Community College . The Effects of Tamoxifen of Rat Embryo Fibroblasts Gwendolyn Steffen—Northern Kentucky University (Sponsored by Ray Richmond) . The Effect of Residency Status upon Site Defense in Plethodon dorsalis Martha Dahlgren and Paul Cupp, Jr—Eastern Ken- tucky University . Molecular Orbitals of 1-Aminocyclopropane-1-Car- boxylic Acid and 2-Aminocyclopropane-1-Caroxylic Acid James L. Meeks and Larry D. Bigham—Paducah Community College . Effect of Ag-Chemicals and Toxic Compounds on Hu- man Erythrocyte Delta Aminolevulinic Acid Dehy- dratase Habiba Afroze Dowla—Kentucky State University . Molecular Geometry of 1-Amino-1-Cyclopropane- PROGRAM, ANNUAL MEETING 79 Carboxylic Acid Jim Meeks—Paducah Community College 26. Thermal 1,3-Dipolar Cycloreversion of Fullerene Isoxazolines Vahid Majidi and Robert Pogue—University of Ken- tucky; Magdalena Poplawska—Warsaw Technical University; Craig Thomas—University of Indiana (In- dianapolis); Mark Meier—University of Kentucky Trans. Ky. Acad. Sci., 56(1-2), 1995, 80-95 ABSTRACTS OF SOME PAPERS PRESENTED AT THE ANNUAL MEETING, 1994 AGRICULTURAL SCIENCES Analysis of esfenvalerate on vegetables. D. GREENE,* G. ANTONIOUS, and M. E. BYERS, Kentucky State University, Frankfort, KY 40601. Esfenvalerate, [(s)-cyano (3-phenoxyphenyl) methyl (s)- 4-chloro-alpha-(1-methyl ethyl) benzene acetate], is the S isomer of the pyrethroid fenvalerate (70% or more active isomer of fenvalerate). An experiment was conducted in 1992, at Kentucky State University Research Farm (Franklin County, Kentucky), to study dissipation of es- fenvalerate on pepper and pumpkin plants following spraying. Field plots (22.0 * 3.7 m, n 160) were ar- ranged on a 10% grade in a randomized complete block design and were planted with green bell pepper and pumpkin. Plots contained 10 rows oriented along the con- tour of the slope. Esfenvalerate was extracted using n- hexane from representative plant samples collected at dif- ferent time intervals for residue analysis. The samples in- dicated initial deposits of 3.34 and 1.18 ppm on pumpkin and pepper leaves, respectively. Only trace levels were de- tected on pepper fruits on day 21 (0.0001 ppm). Half-life values were 1.11 and 2.79 d on pumpkin and pepper fruits, respectively, whereas the values were 1.92 and 3.38 on pumpkin and pepper leaves, respectively. Generally, residues of esfenvalerate on the treated vegetables were low. These low residues are encouraging because risk of exposure to the consumer is low. Dacthal persistence in soil. N. BAKER,* M. E. BYERS, G. F. ANTONIOUS, and D. HILBORN, Kentucky State University, CRS, Frankfort, KY 40601. The use of herbicides to control weeds on erodible lands may reduce the need for tillage and contribute to a sustainable agricultural system. But soil applied herbicides are perceived to potentially affect surface water and groundwater. The fate of a compound in soil is directly related to its potential fate in water. Dacthal (DCPA, di- methyl 2,3,5,6 tetrachloro-1,4-benzenedicarboxylate), a selective herbicide, was applied on 21 April 1994 at 10.1 kg/ha to plots (3.7 X 22 m) on a 10% slope, with Lowell silt loam soil, to which broccoli transplants were planted. Plots in 1994 had fescue strips every row (F1), fescue strips every other row (F2), or conventional tillage (CT) as soil treatments. Dacthal extraction methods, extraction efficiency, and persistence over time in soil were deter- mined. The most effective extraction procedure was using soxhlet and hexane with 2-h refluxing. Analysis was by GC-NPD and GC-MS (301 and 332 m/e). Although ap- plied at relatively high rates, Dacthal was found in soil at concentrations of 0.5, 0.29, and 0.08 ppm on 6 June 1994 in CT, F2, and F1 treatments, respectively. Effects of cultivars and planting dates on biomass pro- duction in soybeans. ASLAM TAWHID* and ELMER GRAY, Department of Agriculture, Western Kentucky University, Bowling Green, KY 42101. 80 Recent concern about the environment has resulted in renewed interest in the use of organic matter in agricul- ture. The objective of the present investigation was to de- termine the effects of cultivar maturity differences and planting dates on biomass production of soybean, Glycine max. The research was conducted on the Western Ken- tucky University Farm in 1993. The experimental design was a split-split-plot with four replications. The three planting dates (2 June, 16 June, and 6 July) were main- plots, the three harvest dates were split-plots, and the five cultivars were split-split-plots. Cultivars and their maturity groupings and areas of adaption are McCall (00, Minne- sota), A2506 (II, Iowa), FFR 561 (V, Kentucky), Perrin (VIII, South Carolina), and Laredo (undesignated matu- rity, forage cultivar). Each experimental unit was 45 m?. Seeds were inoculated and broadcast by hand at a rate of 175 kg/ha and covered by disking. Average biomass pro- duction (oven-dry basis) decreased progressively (2,918, 2,450, and 2,088 kg/ha) for the later planting dates. For planting date 1 and 2, biomass yields increased for suc- cessive harvest dates. However, for planting date 3, bio- mass yields did not differ for harvests 2 and 3. When cul- tivars were compared at the early bloom stage, later ma- turing cultivars produced more biomass. Laredo ‘was con- sistently the highest producer. There were significant interactions involving planting dates, harvest dates, and cultivar. Soybean stands and yields were reduced by in- adequate coverage of the broadcast seed, by insufficient soil moisture at planting, and by competition from John- son grass and pigweed. However, these results indicate that soybean is a good source of biomass production in summer. Elkhorn Creek Monitoring Program: Influence of live- stock on water quality. A. BURKS,* F. S. YOUNG IIL, and M. E. BYERS, Kentucky State University, CRS, Frankfort, KY 40601. The presence of pathogenic bacteria in surface water is indicative of human and/or animal waste being present. In Kentucky, fecal coliforms are continually problematic for surface water. The Elkhorn Creek watershed was iden- tified as being impaired by livestock-originated pollution and sediment. Animals given direct access to surface water bodies or the application of generally improper grazing practices may cause stream-bank degradation and erosion. The current study was originally conceived as an interdis- ciplinary demonstration and research project. It was de- signed to show growers useful technology for precluding or reducing animal access to the Elkhorn Creek, and then to provide the grazers with water through novel alternative sources through best management practices (BMPs). Pre- and post-BMP monitoring have been planned. Results thus far indicate pre-BMP BOD (5-day) values of less than 10 mg for both upstream and downstream, fecal co- liforms ranging from 0 to 1,900 col/dl and generally less than 200 col/dl, average nitrate values near 5 ppm, and PROGRAM, ANNUAL MEETING 8] dissolved oxygen values ranging from 4 to 14 mg/liter. The preclusion of cattle in the future should increase water quality. Environmental effects on Indian mock strawberry. BRI- AN D. LACEFIELD* and ELMER GRAY, Department of Agriculture, Western Kentucky University, Bowling Green, KY 42101. Indian mock strawberry, Duchesnea indica, is a com- mon lawn weed favored by relaxed lawn management. Re- search was conducted to characterize the growth and re- production of individual Indian mock strawberry plants at Bowling Green, Kentucky. The plants were transplanted from 10 different source locations in the experimental area (60 X 60 m) to six test plots in the same experimental area. Each test plot included 10 plants from each source, resulting in 600 plants for the experiment. A test plot con- sisted of a 10 X 10 Latin square design with plants spaced 30 cm apart within and between rows. Individual-plant data were collected on petiole length, leaflet length, and leaflet serrations at transplanting; and on plant height, number of leaves, number and length of stolons, number of rooting plantlets, and number of fruits at intervals dur- ing the growing seasons of 1992 and 1993. Plants from the different sources varied significantly for length of pet- ioles and leaflets and for numbers of serrations. Plant pro- liferation, as measured through the various indexes, varied significantly among test plots. Maximum expressions of the characters for the different test plots included 100% of plants producing stolons, an average of 22.8 stolons/ plant, an average of 491.4 cm stolon length/plant, 99% of plants producing plantlets, an average of 27.7 plantlets/ plant, 98% of plants producing fruits, and an average 11.3 fruits/plant. Indian mock strawberry is a prolific weed through both asexual and sexual reproduction. Growing container herbs in composted materials. TENA M. WRIGHT* and ELMER GRAY, Department of Agriculture, Western Kentucky University, Bowling Green, KY 42101. An increased interest in substituting recycled nutrients for chemical additives in gardening was the basis for this research. Growth media consisted of four materials (local soil, brush compost, leaf compost, and N-Viro soil) and 1: 1 mixtures of the materials, resulting in 10 growth-media treatments. Each treatment was repeated four times for a total of 40 containers. Each plastic barrel container (58 cm diameter, 38 cm depth) contained ca. 60 liters of mix- ture. No pesticides or fertilizers were used. Results were reported on days to emergence for borage, summer sa- vory, and thyme; plant material weight for sweet basil; and number of leaves on parsley. For days to emergence, mixtures had no influence on thyme. The range was 7-19 days for borage and savory. Of the mixtures, 50% soil: 50% brush had the longest delay in emergence for borage; 100% soil resulted in the longest delay for savory. Total weight of sweet basil (three plants per container) ranged from 240 to 10,145 g per container. Its highest average weight, 7,180 g, occurred in the mixture 50% leaves : 50% brush; the lowest, 1,573, occurred in 100% brush. The number of leaves produced per parsley plant ranged from 41 to 349 and averaged 119. The highest number of leaves was found in the 100% N-Viro soil, the lowest was in 50% leaves :50% N-Viro soil. When the results were combined for all herbs, no mixture was superior. However, the mixtures containing brush produced the least favorable results. Influence of constructed wetlands on nutrient and pathogen removal. F. S$. YOUNG III,* M. E. BYERS, and G. F. ANTONIOUS, Kentucky State University, CRS, Frankfort, KY 40601. In Kentucky, NPS pollution from on-site sources affects environmental and human health. In response to this problem, in excess of 1,400 on-site constructed wetlands exist in Kentucky. However, few have been monitored. These systems generally consist of a lined trench filled with rock, planted with emergent macrophytes, and with water level maintained below the rock surface. The inlet of the system is fed septic-tank effluent. Macrophytes, their symbionts, and other benthic micro-organisms con- tact the flowage and metabolize nutrients and consume pathogens. Effectiveness of treatment can be determined through monitoring. Acceptable level of treatment is de- termined by regulators. We have monitored four systems for 2 yr. Some of these systems are mature and seem to have reached their biotic potentials. Fecal coliform re- moval efficiencies often approach 99% but usually far ex- ceed 200 col/dl. System’s efficiency regarding nutrient re- moval is variable but generally not effective. System's loads seem to generally exceed capacity. Increasing real residence time and adequate maintenance will likely in- crease efficiency. Influence of incident light intensity and fertilization on growth and development of pawpaw [Asimina triloba (L.) Dunal] seedlings. LAKEASHA JONES,* DESMOND R. LAYNE, and MICHAEL G. KWANTES, Atwood Re- search Facility, Kentucky State University, Frankfort, KY 40601. At KSU we are conducting research to overcome the horticultural limitations to developing pawpaw as a new commercial fruit crop. Currently, no scientifically based recommendations exist for pawpaw seedling propagation. Pawpaw enthusiasts recommend growing seedlings in shade and providing fertilizer once transplanted. The pur- pose of our study was to determine whether supplemental illumination and fertilization could result in more robust seedlings for field transplanting. Stratified pawpaw seeds were planted in a greenhouse and grown until seedlings reached the 2-3 leaf stage. Uniform plants were selected and assigned one of the four following treatments for 5 wk: (1) ambient light plus fertilizer (Amb/+ Fert), (2) am- bient light without fertilizer (Amb/—Fert), (3) shade (80%) plus fertilizer (Shade/+ Fert), and (4) shade without fertilizer (Shade/— Fert). Seedling height and leaf number 82 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) were recorded 3X/vk. Following 5 wk of treatment, plants were destructively harvested to determine biomass accumulation, and leaf samples were collected and ana- lyzed for chlorophyll. Within 15 d of treatment, Amb/ +Fert seedlings were significantly taller and had more leaves than those in the other three treatments. Following 5 wk, the Amb/+Fert seedlings had 40% more biomass than the Amb/—Fert and more than double the biomass of seedlings from either shaded treatment. Unfertilized plants had leaves with significantly less chlorophyll than fertilized plants. Seedlings provided with additional light and fertilizer were significantly larger, had a more devel- oped root system, and were more suitable for transplant- ing than seedlings grown without these additions. Plant (wheat) and soil analyses following high rates of manure applications on corn. RAY E. JOHNSON, BRENT BASHAM,* BRYAN KESSLER, O. W. DOT- SON III, DAVID STILES, and ALVIN BEDEL, De- partment of Agriculture, Western Kentucky University, Bowling Green, KY 42101. The study was conducted on a Pembroke silt loam (fine- silty, mixed, mesic, Typic Paleudalf) on the Western Ken- tucky University Farm. Dairy cow solid manure was added at rates to supply 0, 168, 336, 672, and 1,344 kg/ha of total N, referred to as the 0, 1X, 2X, 4X, and 8X rates, respectively. Liquid swine-manure was added at the 0, 1X, and 2X rates. One plot in each replication received only urea to supply 168 kg/ha of N. Applications were prior to planting of corn. Corn had been planted on these plots and then the whole plant was removed. Wheat was plant- ed in the fall without any further manure application. Wheat dry-matter yields on the cow manure treatments varied from 2,164 kg/ha to 4,842 kg/ha with an increased response of the 2X, 4X, and 8X manure rates (P < 0.05) over the 0, F, and 1X treatments which were not differ- ent. Nitrogen removal via the wheat plant followed the same pattern with the 4x and 8X treatments removing over 60 kg N/ha (P < 0.05) compared to the 27-36 kg N/ ha removed by the wheat on the other treatments. The rates of nutrient uptake and removal tended to follow the same pattern for P (5-15 kg P/ha), K (19-92 kg/ha), S (2.8-7.3), Mg (3.4-13), and Ca (4.8-11.9 kg/ha) with a fairly consistent difference (P < 0.05) with the two higher applications being greater than the 0, F, 1, and 2X treat- ments. Wheat dry-matter yields on the 0, F, 1X, and 2x swine manure application were 2,640" kg/ha, 2,614", 2,774" and 4,203" kg/ha, respectively, for the four treatments. With the swine manure study, the following nutrient re- moval was observed: N, 31-43 kb/ha; P, 6 to 8; K, 37 to 53; and sulphur, 3 to 4 kg/ha. In the cow manure study, soil micronutrient elements varied, with the following av- erage levels observed: S, 13.6 ppm; B, 0.82 ppm; Cu, 2.56 ppm; Fe 28.5 ppm; Mn 166 ppm; and Zn, 5.6-6.1 ppm for 4X and 8X treatments (P < 0.05) down to 3.5-2.5 ppm for lower levels of manure application. It would ap- pear that the summer corn crop removed a considerable amount of animal waste nutrients on the F, 1X, and 2X treatments as increased forage production and nutrient removal were noted mainly on the 4X and 8X treatments. Street tree inventory: putting the data to work. JAMES M. MARTIN, Department of Agriculture, Western Ken- tucky University, Bowling Green, KY 42101-3576. Data have been collected and evaluated from the in- ventory of ca. 6,000 street trees in Bowling Green, Ken- tucky. Now the data must be used to effect change in the city’s street-tree care and planting program. Six species constituted 50% of the total. Three of these six are inferior species, not suitable for this use. There are 309 trees in need of immediate removal; most of the remaining pop- ulation requires attention and periodic care. Currently no ordinance governs selection and planting of street trees in Bowling Green. To put these data to work, three necessary steps are being implemented. This first is the political/ governmental support from the city to be accomplished in part by direct appeal to the City Commission. The sec- ond is the formulation of policy, including a street-tree ordinance based on these data to be prepared by the City Tree Board. The third step is a public education to make the citizens aware of the problems and the potential. Sustainable agronomic research projects at Western Kentucky University. ELMER GRAY,* ASLAM TA- WHID, TENA M. WRIGHT, and BRIAN LACEFIELD, Department of Agriculture, Western Kentucky University, Bowling Green, KY 42101. The recent sustainable agriculture movement has en- couraged renewed interest in old agricultural practices and has enhanced the attention being given to selected new developments. Agronomic research at Western Ken- tucky University has involved several aspects of sustaina- ble agriculture. Pre-emergence mulching of garden beans with leaf compost increases the percentage of emergence and pod yield. Reinstatement of more intensive lawn clip- ping management aided in control of Indian mock straw- berry and common blue violet, thereby reducing the need for herbicide application. Vegetable and herb production using containers of various compost media has been suc- cessfully demonstrated. Through the use of selected crops (wheat, rye, spring oats, soybeans, and sorghum), it is pos- sible to have a supply of organic matter throughout the year. The use of companion crops as living mulches has received preliminary investigation. Although such crops have shown promise for controlling selected weeds, they have been competitive with the main crop. Treating livestock waste waters with constructed wet- lands. DAVID STILES, BRYAN KESSLER,* O. W. DOTSON III, BRENT BASHAM, RAY JOHNSON, and ALVIN BEDEL, Department of Agriculture, Western Kentucky University, Bowling Green, KY 42101. A commercial producer of swine in south central Ken- tucky has a constructed wetland system to handle part of the waste produced from a 1,500-sow (12,000-15,000 an- imals at any one time) operation. The waste is routed to PROGRAM, ANNUAL MEETING 83 a two-cell-in-series lagoon system and then from there to land application or in summer to a 4 ha wetland system with 9 cells. The first three wetland cells served as mixing ponds where the effluent from the lagoon (2nd) is diluted so that the NH,-N level is ca. 100-150 ppm. The cells are predominantly cattail (Typha latifolia), bulrush (Scirpus validus), panicum (Panicum sp.), and reed (Phragmites australis). The following data, collected in summer 1994, represent the average of the first three cells and the % removal through wetland cell 9 in the closed system: am- monia-N 59, mg/liter, 93.7%; total P, 16.8 mg/liter, 72%; total suspended solids, 504 mg/liter, 21%; total dissolved solids, 715 mg/liter, 23%; fecal coliforms, 33,000 CFu, 100 ml, 85%; and BOP 92, mg/liter, 56%. Dissolved oxygen increased almost three-fold through the cells. Constructed wetlands appear to be an effective and reliable means for treatment of swine lagoon effluent. Further studies of management and monitoring are indicated to follow the start-up year of functioning of these wetlands. Using composts for growing vegetables in containers. ASLAM TAWHID* and ELMER GRAY, Department of Agriculture, Western Kentucky University, Bowling Green, KY 42101. Container gardening provides the urbanite the oppor- tunity to grow vegetables in minimum space. The use of composted waste materials adds to the attractiveness of container gardening. Our study was conducted to evaluate common waste products as soil amendments for growing food crops in containers. The materials included soil (S), leaf mulch (L), wood mulch (W), N-viro soil (N), and chicken manure (C). Each of the 40 plastic barrel sections (58 cm diam, 38 cm depth) contained ca. 60 liters of one of the following mixtures (percent by volume): S(100); S(50) L(50):; S(50): W(50); S(50) L(25) W(25); S(50) L(25) C(25): $(50) W(25) C(25); $(50) L(25) N(25); and S(50) W(25) N(25). The mixtures not including chicken manure (C) or N-viro soil (N) were supplemented with 56.0 N, 24.5 P, and 46.5 K kg/ha. The 100% soil was used as the standard for evaluating the mixtures. Fifteen different food crops were grown in these mixtures in 1993. Seedling emergence, seed or fruit yields, plant height, and biomass productions indicated that some mixtures were equal or superior to soil only. For example, bush snap beans had higher emergence and pod yield on SLN than on S. There were many examples of interactions between mixtures and crops and mixtures and cultivars within crops. These re- sults indicated that gardening with waste compost has strong potential for urban gardeners. Vegetable culture on erodible land. D. HILBORN,* M. BYERS, and G. ANTONIOUS, Kentucky State Univer- sity, Frankfort, KY 40601. Limited-resource farmers are looking for profitable al- ternative crops. Broccoli is a possible alternative. Past studies, with other vegetables on erodible land (10% slope), have shown that fescue strips are effective in re- ducing water and soil runoff. Our study involved three soil treatments for comparison: (1) no mulch or conventional till (CT), (2) fescue every other row (FEOR) of crop, and (3) fescue every row (FER) of crop with respect to effect on broccoli yield. Spring and fall broccoli were transplant- ed (17,857 plants per hectare) and eventually harvested; results for the year were combined. Though head mass and diameter on any given harvest date were not signifi- cantly different, overall head mass and diameter were sig- nificantly higher in the conventional till plot. Head mass (g): CT = 158.7, FEOR = 140.6, and FER = 144.0 with head diameter (mm): CT = 98.2, FEOR = 91.3, and FER = 90.2. With respect to total annual harvest, the conven- tional till plot produced the highest annual yield of 2,700 kg/ha compared with 2,497 kg/ha for the fescue-every-row plot and 2,434 kg/ha for the fescue-every-other-row plot. Therefore, the conventional till plot yielded a better over- all harvest in kg/ha, but not significantly enough to offset the benefits from fescue, which helped to retard loss of water and soil. This study bears repeating at a higher planting density. BIOCHEMISTRY AND PHARMACOLOGY Apoptosis in differentiated and undifferentiated murine F9 teratocarcinoma cells. E. DEATON,* S. VOGEL- POHL, B. BALDWIN, J. CARTER, and R. SNYDER, Wood Hudson Cancer Research Laboratory, Newport, KY 41071. Apoptosis, a genetically driven process wherein cells commit suicide in response to internal and external stim- uli, is fundamental in embryological development and in maintenance of organ homeostasis in multicellular organ- isms from nematodes to man. Aberrant modulation of the apoptotic process is associated with tumor progression, neurodegenerative disease, and other human conditions. As part of our ongoing program aimed at understanding the molecular basis for, and possible modulation of, this process as it relates to cancer, we have investigated spon- taneous and induced apoptosis in pluripotent murine F9 teratocarcinoma cells that had or had not been stimulated to differentiate into endodermal cells by 72 hr exposure to 1 pM retinoic acid (RA). Apoptosis was induced by the S-phase specific agents, ara-~C and etoposide and the non S-phase agent, hydrogen peroxide. Background levels of apoptosis were taken to represent spontaneous rates. Apoptotic DNA fragmentation was measured by DNA/ Dye binding assay and agarose gel electrophoresis. Mor- phometric analysis of stained cells was also used both to verify the apoptotic nature of the event and to analyze death on a per cell basis. We have determined that F9 cells can be induced to undergo apoptosis by all three agents and that this cannot be antagonized by agents such as cycloheximide or zinc, which are classical inhibitors of the process in most cell types. RA-differentiated F9 cells display a level of spontaneous apoptosis similar to that seen in undifferentiated cells. However, differentiated cells exhibit clearly different responses to agent-induced apoptosis than do undifferentiated cells. These findings 84 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) were discussed with regard to their implications in cancer therapy. BOTANY AND MICROBIOLOGY Cryptococcus neoformans serotype groups found in clinical and environmental isolates. JOHN M. CLAU- SON* and LARRY P. ELLIOTT, Department of Biology, Western Kentucky University, Bowling Green, KY 42101- 3576. Cryptococcus neoformans is the most life-threatening fungal pathogen that infects patients with the acquired immunodeficiency syndrome (AIDS). Locally C. neofor- mans var. neoformans has been associated with pigeon fe- ces in those months with an average temperature of 68.8°F and above. Clinical and environmental isolates of C. neoformans obtained from area hospitals and the en- vironmental sampling, respectively, have been grouped into their variety status utilizing Canavanine Glycine Bromothymol Blue agar. Antisera against C. neoformans serotypes A, B, C, and D was produced in rabbits and utilized in serotypic characterization. Serological studies utilizing monoclonal antibody against each of the four se- rotypes have been employed. Comparing serotypes found in the environment to those isolated from patients in area hospitals will provide information leading to the origin of infection and ultimately inform immuno-compromised in- dividuals on situations in which exposure to C. neoformans is likely. Dogwoods: molecules, morphology, and relationships. ZACK E. MURRELL, Department of Biology, Western Kentucky University, Bowling Green, KY 42101. The dogwoods (Cornus) have been variously divided or segregated over the past 3 centuries, usually on the basis of misunderstood inflorescence structure or a disregard for natural groups. A morphological analysis of the genus, using macro-morphology, micro-morphology, anatomy, and chemistry, supports the recognition of all nine sub- genera as a single genus. Molecular sequence data derived from the internal transcribed spacer (ITS) region of 18- 26S nuclear ribosomal DNA are difficult to align at the subgeneric level, indicating that splits between the lin- eages are ancient. The blue- and white-fruited dogwoods comprise two thirds of the genus but have been poorly represented in analyses of relationships. Although these dogwoods have generally been considered shrubs, there are four architectural types represented; molecular data from the ITS region support recognition of at least three lineages within the group. The blue- and white-fruited dogwoods have traditionally been segregated into taxa with opposite or alternate leaves, but the chromosome number and endocarp structure of the South American C. peruviana blur this distinction. Comparison of morpho- logical and molecular data supports recognition of all of the blue- and white-fruited dogwoods, except C. oblonga, as subgenus Thelycrania. CHEMISTRY College chemistry—Can we predict which students will be successful? L. C. BYRD and D. R. HARTMAN,* De- partment of Chemistry, Western Kentucky University, Bowling Green, KY 42101. Attempts to identify underprepared students encour- aged investigations into correlation of success in college chemistry with high school: grade-point average, class rank or chemistry grade, ACT, SAT-math, aptitude test scores and/or customized tests to measure chemistry/math skills. Hovey and Krohn, leaders in the investigations, de- veloped the Toledo Chemistry Placement Examination (TCPE). Several universities require TCPE raw scores above 35-50 for entrance into college chemistry. Corre- lations of success in college chemistry to various param- eters found ranges from 0.21 to 0.51 with multivariate analysis giving the best correlations (0.59-0.63). The De- partment of Chemistry at Western Kentucky University (WKU) determined correlation coefficients for each ACT score, high school grade-point average or TCPE raw scores to success in college chemistry (grade of A, B, or C). The best correlation (0.55) was with the TCPE total. The next best correlation (0.49) was with the TCPE, part I (math section). To help students be more successful and to give them a better experience in chemistry at WKU, the TCPE is given prior to registration; the scores are used to place students in college chemistry + math, pre- paratory chemistry + math, or no chemistry—math only. In fall 1994, 514 students took the TCPE; 215 were en- rolled in college chemistry, 85 in preparatory chemistry, and 214 took no chemistry. In fall 1993, 338 students en- rolled in college chemistry with 175 remaining after mid- term, and 75 enrolled in preparatory chemistry. GEOLOGY A conceptual model for the occurrence of hydrochem- ical facies in an unmined area of the Eastern Kentucky Coal Field. DAVID R. WUNSCH, Kentucky Geological Survey, University of Kentucky, Lexington, KY 40506- 0107. A comprehensive hydrogeologic and hydrogeochemical study was conducted at an unmined site in the Eastern Kentucky Coal Field. Sixteen piezometers were installed to approximate a vertical grid in a ridge characteristic of the geologic and hydrologic conditions of the region. Pi- ezometer placement was based on data collected from geologic core description, geophysical logs, water-injection packer tests, and down-hole camera investigations. Water levels were measured for a period of 1 yr on approxi- mately 10-day intervals. Water samples were collected monthly to evaluate temporal variation in water chemistry. A one-time sampling for tritium analysis was performed to aid in determination of recharge zones. Piezometers monitoring shallow, fractured bedrock and coal beds showed the greatest temporal water-level fluctuation and hydrochemical variation. These effects decreased with depth below the surface. Coal beds of the Hazard series PROGRAM, ANNUAL MEETING 85 apparently act to dewater the upper portion of the ridge by laterally transmitting water, which discharges as springs or seeps. Interpretation of the tritium data indicates that most of the ground-water recharge enters the ridge along the hillslope where fractures, along with a reduction in the degree of slope, allow for the greatest infiltration of ground water. Temporal variation in ground-water chem- istry was minimal at the site except for a few specific cases. Ground water derived from coal seams contained the low- est pH and was predominantly a Ca-Mg-HCO, water type. Water derived from fractured zones varied between Ca-HCO, and Mg-SO, water types. Ground water from the ridge interior was a Na-HCO, type, which contained a high pH and characteristically high fluoride. Barium was found in anomalously high concentrations (>1.0 mg/L) in piezometers near the discharge area (valley bottom) where Na-Cl] water types were encountered, although ground water with elevated barium concentrations was also shown to exist in other locations. Sulfate reduction and cation exchange appear to control the occurrence of barium. Re- action path modeling of the geochemical evolution of ground water at the site showed excellent agreement with observed trends. A set of plausible water-rock reactions is given for the occurrence of each hydrochemical facies zones within the ridge. Computer-aided instruction in geology using photo- graphs transferred to CD-ROM. DAVID R. DOCK- STADER, Division of Natural Sciences and Mathematics, Jefferson Community College, Louisville, KY 40202-2005, Establishment of a standard for Photo CDs has allowed inexpensive commercial transfer of photographic nega- tives and slides to digital format on CDs. The standard Kodak Photo CD contains a photograph in five levels of resolution. The 384 X 256 files are adequate for comput- er-aided instruction. At this resolution, the file for each photograph occupies 232 kilobytes of memory and may be readily transferred to a disk drive for use on computers that do not have a CD ROM drive attached. Use of dig- itized images from CD ROM offers several advantages over use of original photographs. Among these advantages are preservation of original photographs, rapid random and repeat access, inexpensive duplication without loss of quality, and ease of incorporation into computer-driven instructional media. Using funding from a UKCCS Mini- Grant, I have transferred 500 slides to Photo CDs. They include photographs of the Falls of the Ohio, Clifty Falls State Park, Red River Gorge, Niagara Falls, Glacier Na- tional Park, Arches National Park, the effects of Hurricane Hugo, and many other geologically interesting areas. Cop- ies of these photographs are available for the cost of du- plicating and shipping the disks. Some of the photographs have been incorporated into illustrated computer instruc- tion and simulated field trips for student use with HyperCard on the Macintosh computer. Copies of these HyperCard materials may also be obtained from me. Effect of agricultural chemicals on ground-water quality in lacustrine deposits of the Western Kentucky Coal Field. PHILIP G. CONRAD* and JAMES S. DINGER, Ken- tucky Geological Survey, University of Kentucky, Lexing- ton, KY 40506-0107; LYLE V. A. SENDLEIN, Kentucky Water Resources Research Institute, Lexington, KY 40506-0107. Ground-water quality at two farm sites in western Ken- tucky was monitored before and after application of ag- ricultural chemicals. Both sites lie in wide, flat valley floors of the Western Kentucky Coal Field. Surface sediments at each site consist of Pleistocene lacustrine deposits and some alluvial deposits and are underlain by Pennsylvanian sandstone, shale, coal, and limestone of the southern II- linois Basin. Shallow ground-water flow above homoge- neous clays was mostly lateral toward local and regional- scale drainage ditches at both sites. Nitrate-nitrogen con- centrations at the Hopkins County site were periodically over 20 mg/liter in ground water less than 10 ft in depth. Conversely, nitrate-nitrogen concentrations at the Daviess County site were all below 1.5 mg/liter from the water table to over 60 ft in depth. The large difference is ap- parently caused by slower denitrification at the Hopkins County site due to lower concentrations of organic carbon below the water table than in Daviess County. Organic carbon and other nutrients are necessary for aggressive denitrification by bacteria. Near the Hopkins County wells, the addition of nitrogen fertilizer raises the concen- tration of nitrate-nitrogen in shallow ground water. The vertical movement of ground water and pesticides was im- peded by argillaceous lacustrine deposits. Some down- ward flow of very shallow ground water and pesticides was noted where there is little lacustrine clay above bedrock and where the clay is perforated by macropores. Effect of fractures on availability of ground water and occurrence of pesticides and nitrate in the epikarst at a site in the Inner Blue Grass Region, Kentucky. DWAYNE M. KEAGY* and JAMES S. DINGER, Kentucky Geo- logical Survey, University of Kentucky, Lexington, KY 40506-0107; STEVEN K. HAMPSON, UK Federal Fa- cilities Oversight Unit, Kentucky Water Resources Re- search Institute, University of Kentucky, Lexington, KY 40506-0107; LYLE V. A. SENDLEIN, Kentucky Water Resources Research Institute, University of Kentucky, Lexington, KY 40506-0107. A study site in the Inner Blue Grass Region is charac- terized by gently rolling topography, sparse outcrops, and scattered, broad, shallow sinkholes. Bedrock consists of interbedded limestones, siltstones, and shales. Soils are generally clay-rich loams. Agricultural practices include growing corn, soybeans, and tobacco and pasturing beef cattle. Shallow ground water in the epikarstic zone was the major focus of this study. Two sites were chosen: a cattle pasture where no agricultural chemicals were ap- plied and a corn field where atrazine, metolachlor, and nitrogen fertilizer were applied. The sites were then sub- divided into fracture zones and interfracture zones. Wells located in fracture zones produced more shallow ground 86 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-2) water than those located in interfracture zones. Triazines and metolachlor were found only at the crop site, whereas nitrate, found in higher concentrations at the crop site, was detected at both sites. In fracture zones, triazines and nitrate concentrations showed little variation with depth and were lower than in shallow interfracture zone ground water. Triazines and metolachlor concentrations in both fracture and interfracture zones peaked ca. 2 wk after ap- plication and then fell to below 1 wg/L less than 3 months later. Pesticides and electrical conductivity measurements indicate that shallow epikarstic ground water from inter- fracture zones discharges into solution conduits, fracture networks, and eventually from springs. Drilling wells in fracture zones in similar geologic settings should produce the best water quality and provide better ground-water yields. Geology of roof falls in underground coal mines: com- parison between eastern and western Kentucky. DAVID K. HYLBERT, Department of Physical Sciences, More- head State University, Morehead, KY 40351. The geology of rocks associated with coal beds is known to have a direct bearing on the stability of roof rocks in underground coal mines. Kentucky is unique in that the eastern Coal Field is part of the Appalachian Basin and the western Coal Field is part of the Illinois Basin. The kind and geometry of rocks in eastern Kentucky are as- sociated with sediment deposition in a basin with relative- ly unstable and more rapidly subsiding platforms. The rate of sediment influx was also relatively greater. The resulting coal beds and associated rocks are typically more non- marine in their character. Roof falls in these rocks are most often attributed to sandstone channel-fills, non-tec- tonic faults, and slips caused by differential compaction, lateral facies changes, and geologic structures such as ket- tlebottoms and concretions. Coal beds and associated rocks deposited in the Illinois Basin typically reflect de- position on more stable platforms with relatively less sed- iment influx. The rocks show more of a marine influence. In addition to roof falls caused by sandstone channels and non-tectonic faults caused by differential compaction, an unstable roof may result from clay dikes or clay-dike faults. Also, tectonic faults in well-defined systems are much more numerous in western Kentucky. Nonpoint-source pollutant loads in a karst aquifer un- derlying an intensive-use agricultural region, Kentucky. JAMES C. CURRENS, Kentucky Geological Survey, Uni- versity of Kentucky, Lexington, KY 40506-0107. The Pleasant Grove Spring Basin, southern Logan County, Kentucky (10,082 acres, 40,823 he), was studied because it is largely free of non-agricultural pollution sources. Water samples were collected at six locations over a 14-month period. Pleasant Grove Spring was instru- mented to monitor five water-quality parameters, stage, and discharge velocity, and to automatically collect sam- ples. Stage recorders were installed at four upstream sites. Nitrate is widespread and persistent but concentrations do not exceed drinking water standards (MCLs). Atrazine was detected year round; other pesticides occur. Triazines (including atrazine) exceeded MCLs during spring floods. Bacteria counts always exceeded MCLs. Suspended sed- iment was exceptionally high when storms occurred be- fore cover crops were established. Soil, sand, and gravel hydrostratigraphic units associat- ed with glacial lake deposits, carbonates, and uranium- bearing black shales of Jefferson County area, Kentucky. GRAHAM HUNT, Department of Geography & Geosci- ences, University of Louisville, Louisville, KY 40292. Because the terrain in and around Jefferson County, Kentucky, is estimated to have a high geologic radon po- tential of greater than 4 picocuries per liter, a geologic radon study may be warranted under three geologic in- vestigations: (1) source or generation of radon, (2) migra- tion or transmission of radon, and (3) concentration or trap of radon. During the past decade a random sampling of indoor radon was carried out by mainly government workers in scattered localities of Kentucky, including the study area. The counties were assigned to zones with pre- dicted average radon indoor screening levels; Jefferson County is given a high rating of radon potential. These zone ratings were determined by assessing the following data: indoor radon measurements, geology, aerial radio- activity, soil parameters, and foundation types. Based on a preliminary geologic study it may be concluded that there are many important geologic and climate-controlled features that may have contributed to the source, migra- tion, and concentration of radon in the study area. Inves- tigated rock types that may produce anomalous radon lev- els are (1) black marine shales of mainly Devonian age, (2) karst carbonates, (3) soils, sands, and gravels of Recent to Quaternary age, and (4) glacial lake deposits. There may be localized concentrations of radioactive minerals in or near fractures of high permeability to allow migration and a trapping mechanism of radon. The Kentucky Groundwater Data Repository. BART DAVIDSON, Kentucky Geological Survey, University of Kentucky, Lexington, KY 40506-0107. The Kentucky Groundwater Data Repository, initiated in 1990 by the Kentucky Geological Survey under man- date from the Kentucky State Legislature (KRS 151:035), was created to archive and disseminate Kentucky ground- water data collected by various state agencies and other organizations. Data for 443 water wells drilled in Mc- Cracken County are currently available. In addition, data are also available for drainage areas of tributaries to the Ohio River. The complete data base contains information for over 27,000 water wells, 450 springs, 300 dye traces, and 17,000 water-quality analyses. Types of computerized data in the repository include general water-well infor- mation such as location, usage, total depth, and static wa- ter level; well-construction information; water-quality data such as major and minor ionic constituents, physical prop- erties, isotopic analyses, trace-organic analyses, and bac- PROGRAM, ANNUAL MEETING 87 terial analyses; spring data; discharge measurements; and groundwater dye-trace data. The repository also contains many reports on groundwater topics and maps showing various types of groundwater information. Products and services of the repository include well searches within a user-specified radius of a site location; overlay maps show- ing well locations (7.5-min quadrangle); hard-copy print- outs of groundwater data; groundwater data downloaded to magnetic media (diskette, 9-track tape, 8 mm tape); and assistance with public service requests concerning ground- water. GEOGRAPHY Deaths and injuries from lightning in Kentucky: analysis by location. MARY M. SNOW, Department of Geography and Geology, Western Kentucky University, Bowling Green, KY 42101. Analysis of locations of occurrences of deaths and in- juries caused by lightning in Kentucky was conducted to determine whether a disproportionate number of events was farm related. Relevant data from the National Oce- anic and Atmospheric Administration (NOAA) extend from 1959 to 1992 with the strikes organized by counties. NOAAs publication Storm Data provides details concern- ing specific locations. Each death and injury during the 33 yr was reviewed and further categorized to investigate possible patterns of locations in Kentucky. NOAA's clas- sification of lightning-caused deaths and injuries in the U.S. does not include a category for farm-related events. However, my analysis reveals that 23% of the injuries and 41% of the deaths from lightning in Kentucky took place on farms. Solstices and daily normal maximum and minimum temperature lags. RICHARD K. SNOW, Department of Geography and Geology, Western Kentucky University, Bowling Green, KY 42101. Differences between dates of solstices and daily normal maximum and minimum temperatures were plotted as is- ochrones from 1885 to 1925 and from 1948 to 1992, com- paring seasonal temperature lags as well as the relation- ship to elevation and latitude. Our study focused on Ken- tucky, including stations from 35 to 41°N and from 82 to 94°W. Scattergrams and a Pearson’s product-moment cor- relation coefficient analysis indicated that there was no association between seasonal lag and elevation in Ken- tucky. However, a regression analysis determined that there was a positive relationship between latitude and lag in winter and an inverse relationship in summer. The mean number of days from the. solstices to the daily nor- mal minimum and maximum temperatures has decreased with daily normals slightly slower in winter and slightly higher in summer. HEALTH SCIENCES Effect of ag-chemicals and toxic compounds on human erythrocyte delta-aminolevulinic acid dehydratase. H. A. DOWLA,* M. PANEMANGALORE, M. E. BYERS. and R. MUKHERJEE, Water Quality and Toxicology Pro- gram, Atwood Research Facility, Kentucky State Univer- sity, Frankfort, KY 40601. | Delta-aminolevulinic acid dehydratase (ALAD), a sulf- hydryl enzyme found in human erythrocytes (RBC), cat- alyzes condensation of two moles of aminolevulinic acid (ALA) to form one mole of porphobilinogen. This enzyme is sensitive to inhibition by cadmium (Cd) and some ag- riculture chemicals. We investigated the effect of chemi- cals used in tobacco fields on changes in human RBC ALAD activity in vitro. ALAD was assayed in RBC using ALA as substrate in the presence of dithiothreitol. The data show that % inhibition obtained varied for Cd, ace- phate, maleic hydrazide (MH) and nicotine. Dose re- sponse curves were developed for each chemical; maxi- mum inhibitions obtained for these chemicals were (1) 57% for 2.3 4M of Cd, (2) 52% for 180 mM of acephate, (3) 90% for 6.7 mM of MH-30, and (4) 87.6% for 162.24 mM of nicotine. These data clearly illustrate that RBC ALAD is sensitive to and is inhibited to various degrees by these chemicals. ALAD thus has the potential to be a biomarker to evaluate exposure and extent of toxicity in occupationally exposed tobacco-farm workers. (USDA CBG #93-38814-8733). MATHEMATICS Parts from Dirichlet. JAMES B. BARKSDALE JR., Department of Mathematics, Western Kentucky Univer- sity, Bowling Green, KY 42101. This expository presentation highlights an observation by L. M. Graves [Amer. Math. Monthly 38:277-278] re- garding a logical link between two well-known and fun- damentally significant results that are central items in el- ementary calculus courses. The presenter of this paper suggested that such observation could serve as an element of instructional enrichment for beginning calculus courses. In the note by Graves, it was observed that the procedure of integration by parts is a special instance of the Dirichlet Formula for interchanging the order of integration in an iterated integral; more precisely, the formula of Dirichlet is given by b x b b | | hix, t) dt dx = | | h(x, t) dx dt. In this paper, the implication of “parts” from “Dirichlet” was noted and detailed. The presenter then offered the following direct corollary which, regarding the observation by Graves, replaced the implication with a biconditional. More exactly stated: Let h(x, t) = g(x)f'(t), and let g and f each be Riemann integrable on [a, b], and define G(x) (DF) ‘= G(b) + * o(t) dt; it then follows that b b b i fix)g(x) dx = fix)G(x)| — i f (x)G(x) dx. (DF) 88 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) MOLECULAR AND CELL BIOLOGY Effect of ozone exposure on antioxidant vitamins in fruit tissue. K. M. KUTE,* C. ZHOU, and M. M. BARTH, Nutrition and Food Sciences Department, Bio- logical Sciences, and Graduate Center for Toxicology, Uni- versity of Kentucky, Lexington, KY 40506-00541. Once strawberries are harvested, the length of time which they can be stored is limited by rapid fungal decay and increased rate of senescence. Ozone (QO,), ,), a strong gaseous oxidant, has been demonstrated to reduce fungal growth and abate ethylene produced by the plant, thereby slowing the rate of senescence when used in conjunction with coldroom storage. Despite these benefits, ozone has the potential to react with important antioxidant vitamins in the fruit tissue, thus reducing the antioxidant vitamin content in the human diet. The purpose of this study was to assess the effects of ozone exposure on the antioxidant vitamin content in strawberries through evaluation and quantification of vitamin A and vitamin C activity using traditional spectrophotometric and fluorometric assays (respectively) and High Performance Liquid Chromatog- raphy (HPLC) techniques. Vitamin A activity in strawber- ries was measured using HPLC. Levels of activity were so low in the control group of this cultivar that no meaningful comparison could be made; focus was thus placed on mea- surement of vitamin C activity. Microfluorometric analysis of total ascorbic acid activity (TAA) revealed initial con- centrations of 7.47 mg/g dry weight. At 24 hr, 0.3 ppm berries showed greater vitamin C content than the con- trol, but by 1 wk no significant differences were observed. Although overall quality appeared greater in ozone treated samples, ozone storage resulted in no significant decrease in antioxidant vitamin content over | wk storage time. Effects of teratocyte secretory products on protein syn- thesis by fat body in various larval stages of Heliothis vi- rescens. LISA McGRAW* and DOUGLAS L. DAHL- MAN, Department of Entomology, University of Ken- tucky, Lexington, KY 40508. The tobacco budworm, Heliothis virescens, is a major tobacco pest notoriously resistant to pesticides. Parasitism of H. virescens by Microplitis croceipes, a braconid wasp, interferes with development of the budworm and even- tually kills it. As the parasitoid egg hatches within the host, the serosal membrane dissociates into individual cells called teratocytes. Teratocyte secretory products (TSP) have been shown to inhibit protein synthesis by fat body in H. virescens. The effects of TSP on various larval stages of H. virescens (D1, D2, BD, and CF1) are determined by using an in vitro fat-body assay. About 5 mg of fat body is taken from various staged larvae and labeled with *°S Methionine. Four larval equivalents of TSP are then add- ed and the samples are incubated for 4 hr. After incuba- tion, the samples are washed in various amounts of tri- chloroacetic acid (TCA) to precipitate the protein. The TCA is removed through ethanol and ether washes. Fi- nally, a SDS/sodium hydrozide wash is used to digest the proteins. The amount of protein secreted by the fat body is measured through the use of scintillation spectroscopy. In assays run without the addition of TSP, early stages (D1 and D2) were shown to synthesize more proteins than later stages (BD and CF 1). When TSP was added to the assay, it inhibited protein synthesis more in D2 stage lar- vae than in the other, later stages. Fatty acid effects on vitamin E levels in cultured en- dothelial cells. BRAD MIDDENDORF* and BERN- HARD HENNING, Department of Nutrition and Food Science, University of Kentucky, Lexington, KY 40506- 0054. One of the first steps in the atherosclerotic disease pro- cess is the formation of lesions within the vascular lining. Lesion formation may be initiated by the disruption of the vascular endothelium. Different nutrients can play a key role during this process, in both disruption and protection. Fatty acids, especially unsaturated fatty acids, can induce oxidative stress upon endothelial cells. Linoleic acid [18: 2(n-6)| has been shown to be cytotoxic to endothelial cells. Oxidative stress can be lessened by the quenching effects of antioxidants such as vitamin E, the most significant lipid soluble antioxidant. Thus, a decrease or alteration in vi- tamin E levels should be expected in endothelial cells ex- posed to linoleic acid as opposed to cells left untreated. To test this hypothesis, an experimental group of cultured porcine pulmonary endothelial cells was exposed to media containing 90 mol/liter linoleic acid for 24 hr. The con- trol group was treated with the same media minus the linoleic acid. Results calculated from HPLC analysis showed no significant difference in overall vitamin E lev- els between the two groups. There were, however, signif- icant differences in vitamin E profiles. Vitamin E (to- copherol) exists as several isomers, differing in degrees of antioxidant properties. The y-tocopherol levels in the treated groups were significantly higher than those in the control group [0.667 to 0.360 nM/mg protein]. These re- sults indicate that exposure of endothelial cells to fatty acids can alter the overall cellular vitamin E profile. This may have important implications in understanding the mechanisms by which dietary fat may influence the ath- erosclerotic disease process. How do lymphocytes recognize antigens in immune privileged sites? WAI KHAN LOH* and JEROLD WOODWARD, Department of Microbiology and Im- munology, University of Kentucky, Lexington, KY 40506. The objective of this summer project was to determine how lymphocytes recognize antigens in immune privileged sites. Spleen cells from normal and transgenic mice were used. In a “naive” mouse that has not encountered an antigen, the frequency of T-cell precursor specific for the antigen is about 1/1,000. If the mouse is exposed to the antigen, the frequency of T-cell precursor will increase. T-cell recognition of the antigen on a target cell initiates differentiation of T-cell precursor to CTL (cytotoxic T- lymphocyte), which then kills the target cell. CTL-pre- cursor Frequency Assay was carried out to measure CTL- PROGRAM, ANNUAL MEETING 89 precursor frequencies of target cells from normal BALB/ C mice and transgenic B6.D* mice that expressed class I MHC antigen, H-2D". If the T-cell precursors in the transgenic mice were exposed to the alloantigen, they would demonstrate a higher CTL-precursor frequency than that of the normal mice. The responding cells in the assay were taken from the normal C57BL6 mice, which did not carry the antigen. The experimental data were insufficient to answer the proposed question. However, they did indicate that the CTL-precursor Frequency Assay was a feasible method to be used to help achieve the ob- jective of this project in the future. Influence of antioxidant and temperature treatments on lipid peroxidation and senescence in broccoli florets. S. M. TAM,* H. ZHUANG, and M. M. BARTH, Biological Sciences and Nutrition and Food Science Departments, University of Kentucky, Lexington, KY 40506-0054. As plant products senesce during the postharvest peri- od, membrane lipids break down resulting in release of polyunsaturated fatty acids (PUFA). Peroxidation of PUFA by lipoxygenase (LOX) results in formation of hy- droperoxides, which contribute to increased cell mem- brane deterioration, hence promoting loss of protein and chlorophyll. Studies have shown that application of anti- oxidants along with reduced temperature storage delay green color loss in broccoli florets. The purpose of our study was to assess the effect of an antioxidant (commer- cial product) and temperature storage treatments (2°C, 10°C, 20°C) on lipid peroxidation and senescence in broc- coli tissue by measuring thiobarbituric acid reacting sub- stances (TBARS), PUFA, C,-aldehyde formation and total chlorophyll loss. There was an increase in TBA values and reduction in chlorophyll content and PUFA as storage temperature increased over time. By 96 hr, TBA values increased 50% in 20°C vs. 10°C samples and by 144 hr, TBA values increased 30% in 20°C vs. 10°C samples. A 3-fold loss in chlorophyll content as observed in 20°C sam- ples by 96 hr and 5 fold loss by 144 hr compared to 2°C samples. Relative PUFA decreased by 30% in 20°C vs. 10°C samples by 144 hr, but no significant difference was observed in 10°C vs. 2°C samples. Application of the com- mercial antioxidant preparation inhibited C,-aldehyde for- mation at the early stage of postharvest storage. Hexanal content in antioxidant-treated samples decreased dramat- ically by 24 hr at 2°C compared to control, whereas hex- enal levels in antioxidant-treated broccoli decreased by 50% compared to control by 72 hr. By 144 hr, hexenal levels were similar among all treatments. These results indicate a direct relationship between lipid peroxidation and senescence. Influence of methyl jasmonate on lipid peroxidation and deterioration in broccoli. D. $. GUEORGUIEVA,* H. ZHUANG, and M. M. BARTH, Biological Sciences De- partment and Nutrition and Food Sciences Department, University of Kentucky, Lexington, KY 40506-00541. Methyl jasmonate (MJ) is a fatty-acid compound, nat- urally occurring and widely distributed in plants. Research studies have shown that exposure of plant leaves to MJ results in increased lipoxygenase (LOX) gene expression, lipid peroxidation (LP) and hydroperoxides formation. Hydroperoxides can cause loss of cell membrane integrity, proteins, and chlorophyll. Thus LP is thought to be re- sponsible for senescence of plant tissue. The aim of our project was to elucidate the mechanism for induction of senescence in broccoli following MJ exposure. The exper- imental procedure involved measuring LOX protein levels and thiobarbituric acid-reactive substances (TBARS) to in- dicate the level of LP and measuring chlorophyll to assess senescence. LOX protein levels were characterized by Western blot. Our results showed that broccoli LOX pro- tein increased during storage. The MJ-treated samples showed higher LOX levels than the controls of the same storage time in all experiments. TBARS significantly in- creased during storage in both controls and MJ-treated broccoli. At 96 hr and especially at 48 hr of storage, the MJ samples demonstrated much greater formation of TBARS than the controls, while at 72 hr certain variation was observed as the MJ samples had slightly lower TBARS levels than controls. Chlorophyll a and chlorophyll b (CB) substantially decreased during storage, particularly Cb, which decreased by 25% from 0 hr to 96 hr in the MJ- treated samples. Total chlorophyll exhibited the same trend, progressively decreasing with time. MJ-treated broccoli had significantly less chlorophyll than controls. These results suggest that MJ enhances LP and will con- tribute to further description of the senescence mecha- nism in broccoli. Interactions of the tobacco vein mottling virus’ cylin- drical inclusion protein with other virus proteins. BETH SMITH* and ARTHUR HUNT, Agronomy Department, N122 Agricultural Science Center North, University of Kentucky, Lexington, KY 40506. The single-stranded RNA potyvirus tobacco vein mot- tling virus contains six classified coding regions: the Help- er Component, CI, Nla, NIb, Coat Protein, and Pl. The Cl gene expresses a protein approximately 70 kDa which has a putative function in viral replication. To better un- derstand the viral protein interactions of the tobacco vein mottling virus the two hybrid system has been employed, which utilizes a unique property of the Gal4 protein. The Cal4 protein has two separable domains: the activation domain and the binding domain. The plasmid pMA424 was genetically engineered to contain the activation do- main; the plasmid pGAD2F was engineered to contain the binding domain of the Gal4 protein. The gene of interest was cloned, creating an in-frame fusion protein with each of the separable domains. The CI was cloned into both plasmids, while clones containing the other genes of in- terest were obtained from the labs of Dr. Hunt and Dr. Pirone. Combinations of both plasmids were transformed into the yeast strain GGY1:171, which has incorporated into its genome the gall-lacZ gene. Beta-galactosidase was then assayed using the chromogenic substrate ortho-nitro- 90 TRANS. KENTUCKY ACADEMY OF SCIENCE 56( 1-2) phenyl beta-D galactoside at OD 421. From this assay, the activity of the gall-lacZ gene was determined. Results from this project will help to better understand the viral protein interactions of the tobacco vein mottling virus. Intracellular niacin status and mechanisms of carcino- genesis. CHRISTOPHER D. WATT* and ELAINE L. JACOBSON, Department of Clinical Sciences, University of Kentucky, Lexington, KY 40536. In the 1960s, researchers discovered that nicotinamide adenine dinucleotide (NAD), the major metabolite of the vitamin niacin, is utilized as a substrate in ADP-ribose transfer reactions. One particular type, the poly(ADP-ri- bose) transfer reaction, is involved in the repair of dam- aged DNA. Subsequent research has shown that a sub- normal intracellular NAD concentration can limit the ef- fectiveness of this DNA repair mechanism. This knowl- edge, plus the fact that DNA damage contributes significantly to cancer development, has encouraged re- searchers to re-examine the relationship between intra- cellular niacin status, mechanisms of carcinogenesis, and cancer incidence. The investigation of this relationship in humans has been assisted by the recent development of a method to assay intracellular niacin status. This technique involves isolation of oxidized forms of the pyridine nucle- otides, NAD and NADP, from primarily erythrocytes. The ratio of [NAD] to [NADP] in erythrocytes is defined as niacin number, a specific measure of intracellular niacin status in humans. The unique metabolism of niacin pre- cursors in humans relative to potential animal models dic- tates the use of humans in this investigation of niacin and carcinogenesis. Assessing the association of intracellular niacin status and cancer incidence was the overall objec- tive of the studies reported here. The development of techniques to conduct this investigation and their appli- cation to the ongoing study of diet and cancer in Malmé, Sweden, will be reported. Understanding the contribu- tions of niacin in limiting carcinogenic cellular events may lead to dietary interventions that could impact cancer pre- vention. Investigation of repressor dependent segregation of plasmid F in Escherichia coli. MICHAEL A. ZGODA* and DONALD P. BIEK, Department of Microbiology and Immunology, University of Kentucky Medical Center, Lexington, KY 40513. I am working with a plasmid that may give some insight into the mechanism by which chromosomes segregate during cell division in prokaryotes. pZC178 contains the partition region from plasmid F and is maintained at a very low copy number, 1-2 per chromosome. The parti- tion region is believed to be an autoregulated operon that expresses two gene products (sopA and sopB) and con- tains a region (sopC) of 12 imperfect 43 base pair repeats that have been suggested to be a functional analog of the centromere of eukaryotic chromosomes. I found that proper partitioning of this plasmid is dependent on lactose repressor binding to a lactose operator site located close to the sopC region. The interesting phenotype observed with this plasmid is that, when the lactose repressor is induced with IPTG, the plasmid becomes actively unsta- ble and is lost at a greater rate than a sop mutant plasmid. My research has shown that some type of anchoring of the plasmid with another cellular component, via sopC, is probably allowing for proper DNA topology. This is es- sential for faithful plasmid maintenance, but the topolog- ical requirements can also be accomplished by transcrip- tion into sopC. The data suggest that, when the sopC re- gion is not able to maintain a specific topology, an aggre- gation of plasmids occurs, accounting for the active instability observed with lactose repressor is induced. Mapping the location of somatostatin-like immunoreac- tive cells in tadpole tectum. ANGIE BAKER* and ELIZ- ABETH A. DEBSKI, Biology Department, University of Kentucky, Lexington, KY 40506. Previous work in this laboratory has shown that so- matostatin-like immunoreactive (Som-ir) tectal cells are distributed non-uniformly in the adult optic tectum: Som- ir cells are predominantly located in the caudal third of the tectum and virtually absent from the rostral third. We wanted to determine whether this non-uniform distribu- tion of Som-ir cells was also found in the developing optic tectum of tadpoles. A stage XVIII Rana pipiens tadpole was anesthetized and then perfused through the heart with saline solution followed by 4% paraformaldehyde. The brain was removed, fixed overnight, and then sunk in a 30% sucrose solution. The tissue was cut sagittally at 16 2m on a cryostat. The anti-somatostatin serum used was obtained from Chemicon and diluted eighty fold. Staining was visualized using a fluorescent secondary antibody. Sec- tions of the optic tectum were divided into regions (cau- dal, middle and rostral) using a grid in the ocular of the microscope. The distribution of Som-ir cells was recorded according to the tectal region and layer in which the cells fell. Som-ir cells were found in all three regions of the tadpole optic tectum. However, most Som-ir cells (49.9%) were located in the caudal third of the tectum with the rest evenly divided between the rostral (26.8%) and mid- dle (23.4%) third. Overall, the lateral, more mature tectal regions did not have any more Som-ir cells than the less mature medial regions. The tectum is a layered structure composed of alternating cellular and plexiform layers. Som-ir cells were distributed fairly evenly between the three cellular layers of the tectum; layer II had 24.0%; layer IV, 26.3%; and layer VI, 19.3% of the Som-ir cells. Many of the remaining Som-ir cells (16.4%) were located in the superficial tectal layers. While adult tectum con- tained an average of 231 Som-ir cells, 647 cells were found in the tadpole tectum. The number and distribution of Som-ir cells in the tadpole optic tectum suggest that somatostatin may be transiently expressed during devel- opment. Its function may be to help in the construction and assembly of the neuronal arrays that subserves visual processing. PROGRAM, Modulation of TNF cytotoxicity by NAD. MANDALA V. WILSON,* DAVID M. HIESTAND, BOYD E. HA- LEY, and ELAINE L. JACOBSON, Department of Clin- ical Sciences, University of Kentucky, Lexington, KY 40536. Tumor Necrosis Factor (TNF) is a cytokine originally named for its ability to selectively kill tumors in several mouse Cell lines. Since its discovery and early testing TNF has been documented to play a role in many cellular ac- tivities, with effects ranging from protective to toxic. Re- cently several cytokines, including TNF, have been shown to selectively bind NAD. Further, preliminary experi- ments showed that addition of NAD enhanced survival of tumor cells in the presence of TNF. To test the hypothesis that NAD can modulate TNF cytotoxicity an alternative assay was needed to measure survival of cells with NAD in the culture medium. Development of these assays will be reported. In addition, experiments designed to study the mechanism by which NAD, TNF, and tumor cells in- teract to modulate the cytotoxic response will be present- ed. Molecular dissection of the mouse a-fetoprotein re- pressor region. CASSANDRA K. BACKER,* AMY W. ELLIS, and BRETT SPEAR, Departments of Microbi- ology & Immunology and Pathology, University of Ken- tucky College of Mariana Lexington, KY 40536-0084. The mouse a-fetoprotein gene (AFP) provides a system to study both positive and negative transcriptional regu- lation. AFP is controlled in a tissue-specific manner, being expressed at high levels in the visceral endoderm of the yolk sac and fetal liver and at low levels in the fetal gut. A dramatic postnatal decline in AFP transcription is seen and within 3 wk after birth the gene is fully repressed. This repression is due to a 550 base pair (bp) region up- stream of the AFP structural gene. Studies in transgenic mice have shown that removing this 550 bp region results in continued transgene expression in the adult liver and gut. Our objective was to further define the region im- portant for AFP shut-off. We hypothesized that negative regulation is controlled by a discrete regulatory motif rath- er than the entire 550 bp region. Standard recombinant DNA methods and the Polymerase Chain Reaction (PCR) were used to obtain three 200 bp subfragments that en- compass the repressor region. These subfragments will be sequenced and subcloned into a eukaryotic expression vector currently being used in our lab. Our long-term goal is to test the repressor activity of these subfragments in transgenic mice. This will enable us to identify the sub- fragment containing the repressor region and further our understanding of AFP repression. These results will help elucidate mechanisms of mammalian gene regulation. Novel synthesis of anti-cancer drugs. LEVI HARPER* and GERALD ROSENTHAL, School of Biological Sci- ences, University of Kentucky, Lexington, KY 40506. Plants must have some way to defend themselves since they are not motile. Many use nonprotein amino acids, ANNUAL MEETING 9] amino acids not normally incorporated into proteins, as a means of chemical defense. Our laboratory focuses on the nonprotein amino acid L-canavanine. L-canavanine is the structural analog of L-arginine characterized by the re- placement of the terminal methylene group with an oxy- gen atom. The terminal oxygen atom, which is more elec- tronegative than carbon, causes canavanine to be more acidic than arginine. Canavanine incorporation into pro- teins in place of arginine causes changes in the protein's R-group interactions thus affecting structure and function. X-ray crystallographic study of canavanine and arginine shows that canavanine is shorter than arginine because oxygen takes up less space than the methylene group. When an extra methylene group is added to canavanine, canavanine should look more like arginine. This greater similarity should facilitate canavanine’s incorporation into proteins at a greater rate. By adding an OH group to a terminal nitrogen or replacing O atom with a sulfur atom, we hope to make canavanine more acidic. Research in rats and nude mice has shown that canavanine causes signifi- cant reduction in tumor size in tumor-bearing rodents. Although canavanine has anti-tumor properties, the tox- icity associated with canavanine caused the treated ani- mals to lose body weight. To remedy this situation, many derivatives of canavanine have been proposed that have greater anti-tumor properties and decreased toxicity. Organelle-specific role of the delta-9 fatty acid desatu- rase. MICHAEL S. SCOWBY,* SERGEI AVDIUSHKO, and DAVID F. HILDEBRAND, Department of Agron- omy, University of Kentucky, Lexington, KY 40546. Genetically engineering plants to produce lower levels of saturated and higher levels of monounsaturated fatty acids is significant for improving these plants’ nutritional values. Such studies are also useful for analyzing the com- plex lipid biosynthesis pathways in plants. Transgenic plants containing a mammalian delta-9 desaturase gene demonstrated significant conversions of saturated to mon- ounsaturated fatty acids in various tissues. Western blot analyses of chloroplasts, mitochondria, nuclei, and micro- somes isolated from leaf tissue of the transgenic plants indicate that the foreign desaturase is associated with the endoplasmic reticulum. Most plant tissues, such as leaves, contain high levels of the 16-carbon saturated fatty acid palmitic acid (>10%), but very low levels of the delta-9 monounsaturated fatty acid palmitoleic acid (<0.1%). The introduced desaturase results in significant accumulation of palmitoleic acid, which provides a qualitative marker for the impact of the desaturase on specific organelle lip- ids and different lipid classes. Palmitoleic acid was found to be incorporated into most major lipid classes found in the isolated organelles, with the greatest amounts occur- ring in the monogalactosyldiacyl glycerol classes. Further organelle and lipid class separations were done to deter- mine the positional specificity of fatty acids w shen they are incorporated into different lipid molecules. Regulation of expression of protease nexin-1 in neural 92, TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) tissue: relationship to Alzheimer’s Disease. PAUL JETT* and STEPHEN ZIMMER, L. P. Markey Cancer Center, Department of Microbiology & Immunology, University of Kentucky Medical Center, Lexington, KY 40536. The molecule protease nexin-1 (PN-1), is a 43,000-dal- ton protein belonging to a family of serine protease inhib- itors known as serpins. PN-1 binds to thrombin and other proteases in glial tissue which may serve as a protective effect in neurodegenerative diseases like Alzheimer's. It has recently been found that the inflammatory mediators Interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-a) are simulators of PN-1. Our laboratory has found that the genomic promoter for PN-1 is stimulated in cells containing ras or eukaryotic initiating factor 4E (eIF-4E). This suggests a potential signaling pathway involving ras, translational control through eIF-4E, and effector func- tions involving IL-1 and TNF-A. These points converge at phospholipase 2 (PLA2), which is the rate limiting en- zyme in precursors of prostaglandins. Clinical studies us- ing inhibitors of this pathway have shown a significant de- lay in the progression of Alzheimer’s disease. These data suggest that PN-1 may be expressed in response to a pro- cess involving neurodegeneration in the brain. Our re- search is focused on testing the validity of this model through the use of techniques such as transient transfec- tions with a PN-1 promoter driven chloramphenicol ace- tyltransferase (CAT) reporter assay in tissue culture and immunohistochemical analysis of Alzheimer tissues. Roles of polyunsaturated fatty acids in neuronal death: arachidonic acid metabolism and lipid peroxidation. MAR- ION STEINER and IRENE HONG,* Department of Microbiology and Immunology, University of Kentucky, Lexington, KY 40536. Alzheimer’s disease is a degenerative neurological dis- order marked by dementia. The roles of polyunsaturated fatty acids in neuronal death were studied as a component in understanding neuronal degeneration in the disease. Classes of polyunsaturated fatty acids that have potent bi- ological effects were examined, namely arachidonic acid metabolites and lipid peroxides. To study arachidonic acid metabolism, PC12 cells (a rat pheochromocytoma derived cell line) were radiolabeled with C-14 arachidonic acid, treated with test reagents such as indomethacin, and then analyzed for metabolites (e.g., prostaglandins) using thin- layer chromatography and paper chromatography. To de- tect products of lipid peroxidation, TBARS (thiobarbituric acid reactive substances) assays were performed. Such techniques were utilized in preparation for the determi- nation of the effects of 8 amyloid peptide on lipid per- oxidation and arachidonic acid metabolism. Analysis of PC12 cells under standard growth conditions (without B amyloid treatment) indicated (1) low levels of production of cyclooxygenase and lipoxygenase pathway products of arachidonic acid metabolism and (2) nondetectable lipid peroxidation. Studies of non-protein amino acid disruption in pro- teins. MICHAEL J. BASS* and G. A. ROSENTHAL, School of Biological Sciences, University of Kentucky, Lexington, KY 40506. Plants must protect themselves from invasion by micro- organisms. Many higher plants defend themselves from predation by synthesizing non-protein amino acids. L-can- avanine, the 2-amino-4(guanidinooxy)butyric acid struc- tural analog of L-arginine, is an example of a non-protein amino acid with demonstrated cytotoxic and insecticidal properties. Canavanine-containing proteins can exhibit an altered three-dimensional conformation. The female mi- gratory locust, Locusta migratoria-migratorioides (Or- thoptera), synthesizes the storage protein vitellogenin for use by developing oocytes. Injection of canavanine into the locust during vitellogenin synthesis results in a re- placement of 18 of the 200 arginyl residues with canavan- ine. This replacement has been shown to alter dramati- cally the native three-dimensional conformation of this protein. Canavanine incorporation into an enzyme can also hinder function. Larvae of the tobacco hormworm, Manduca sexta, synthesize a protective antibacterial pro- tein, lysozyme (EC 3.2.1.17), upon injection of cell wall fragments of Micrococcus lutea. If canavanine is injected into the larvae at the time of bacterial challenge, the ar- ginine analog is incorporated into lysozyme. The newly formed canavanyl lysozyme exhibits a 50% loss in catalytic activity relative to the native protein. Data from the in- sectan proteins isolated from L. migratoria and M. sexta provide evidence that canavanine’s incorporation into pro- tein can alter protein conformation and catalytic activity. However, there needs to be a greater understanding of how canavanine and other non-protein amino acid anti- metabolites function to alter protein conformation. Studies of variation of a surface protein (Szp) of Strep- tococcus equi subsp. zooepidemicus. MATT BLAIR,* JOHN WALKER, and JOHN TIMONEY, Gluck Equine Research Center, University of Kentucky, Lexington, KY 40506. The DNA sequence of the szp genes from a series of 14 S. zooepidemicus strains characterized by Moore and Bryans (MB) has been previously determined; though highly homologous, three regions of variability were iden- tified. The 5’ end of the gene exhibits one of two motifs that were arbitrarily named class I and II. A second region of hypervariability lies 500 bases into the gene. The third region, located near the 3’ end of the gene, translates into a variable number of PEPK repeats in the Szp polypep- tides of the 14 MB strains. To determine whether this structure of the szp gene was present in more recent iso- lates we examined the genetic basis of Szp variation in four strains of Z. zooepidemicus—#54, 54-15, 54-16, and 54-19— isolated simultaneously from the tonsil tissue of a single horse. A Western blot of the surface protein re- vealed three different migration pattems in the four strains, #54 and #54-15 being identical. Using the Poly- merase Chain Reaction (PCR) with specific amplification primers, the entire szp gene and the internal hypervari- PROGRAM, able region of the gene were amplified from each of the four strains. The DNA fragments were then cloned into a plasmid vector and maintained in Escherichia coli. The DNA sequence of the 5’ and 3’ ends of the szp gene as well as of the internal hypervariable region was deter- mined. The results showed that the 5’ region of #54, 54- 15, and 54-19 was like class II and that #54-16 had a class I motif. The hypervariable regions of #54, 54-16, and 54- 19 were different from each other but were identical to sequences of the szp gene found in the MB strains of S. zooepidemicus. The 3’ end of the four szp genes encoded for the same number of PEPK repeats. The DNA se- quence of the szp genes of #54 and #54-15 were identical, which is consistent with the protein profiles. The similarity of DNA sequence of the szp genes of the four #54 strains with those of the Moore and Bryan’s isolates indicated that there is significant conservation of structure in the Szp family of proteins. We also showed that a single horse can carry multiple strains of S. zooepidemicus at the same time. This study may aid in future work of classifying S. zooepidemicus strains. Synthesis of insulin-like growth factor binding protein 2 mRNA in human T-Cells. SANDHYA VENUGOPAL,* LORRI ANN MORFORD, and THOMAS ROSZMAN, Department of Immunology and Microbiology, University of Kentucky, Lexington, KY 40502. Patients with malignant gliomas exhibit a broad im- munosuppression. One explanation for this is that a cir- culating glioma-derived suppressor factor (GSF) interacts with T-cells and renders them unresponsive to antigenic stimulation. Others in the laboratory have developed a monoclonal antibody that initially appeared to reverse GSF-induced suppression in human T-cells in vitro. Based on western analysis of glioma culture supernatants and protein sequencing, the protein recognized by this mono- clonal antibody has been iSISna ee as insulin-like growth factor binding protein 2 (IGFBP-2). However, it is unclear what relationship (if any) exists eee IGFBP-2 and GSF. If IGFBP-2 is inhibitory to T-cell proliferation, one would expect that IGFBP-2 would not be produced by T- cells. The monoclonal antibody to IGFBP-2 reacts with the surface of purified T-cells and can stimulate peripheral blood lymphocytes (PBL) to proliferate in culture. Re- cently it has been suggested that human PBL are capable of producing a number of IGFBPs. To date it is unclear which IGFBPs are actually produced by T-cells, why they are produced, and what role these proteins have in normal lymphocyte function. To examine the synthesis of IGFBPs in T-cells, we utilized Northern blot analysis to determine whether mRNA to IGFBP-2 was actively synthesized in stimulated T-cells. Our data indicate that IGFBP-2 mRNA is synthesized by T-cells. The results demonstrate that GSF and IGFBP-2 are not the same. PHYSICS Comparison of the ultraviolet spectral morphology and MK classifications of B supergiants in the Small Magel- ANNUA L MEETING 93 lanic Cloud. ANTHONY L. BORCHERS* and RAY- MOND C. McNEIL, Department of Physics and Geol- ogy, Northern Kentucky University, Highland Heights, KY 41099. Study of ultraviolet spectra of a sample of B supergiants in the Small Magellanic Cloud is being carried out in col- laboration with Richard P. Fahey and George Sonneborn of NASA Goddard Space Flight Center. As a first step in this study, ultraviolet spectral morphology of program stars is being examined for consistency with published MK spectral classifications. The analysis includes a tabulation of ultraviolet spectral features and their expected variation with spectral type and luminosity class, preparation of spectral sequences of standard stars, and comparison of program spectra with the standard sequences. Standard stars selected for the comparison include B dwarfs and giants as well as supergiants. All spectra are either new or archival spectra obtained with the International Ultravio- let Explorer and processed using the VAX Cluster of the Laboratory for Astronomy and Solar Physics at Goddard. The support of NASA and Northern Kentucky University through the Joint Ventures (JOVE) program is gratefully acknowledged. PHYSIOLOGY AND BIOPHYSICS Effects of nutrients and retention time on the primary production of Kentucky Lake. MARY CHILD,* H. R. KOBRAEI, and B. ANDERSON, Department of Astron- omy and Physics, Murray State University, Murray, KY 42071. Data taken from the Kentucky Lake reservoir system for 1991, 1992, and 1993 were used to calculate experi- mental primary production. The data were also analyzed for possible physical, chemical, and biological factors that could contribute to development of a model for prediction of primary production. Possible correlations were inves- tigated between nitrate/nitrite (NO,/NO,), silicon dioxide (SIO,), soluble reactive phosphorus (SRP), light, temper- ature, carbon assimilation (CAS), and biomass (chloro- phyll a + phaeophyton a). CAS and biomass have a linear relationship to each other and can therefore be used to obtain an experimental value for primary productivity. All available nutrients appear to have the same characteristic behavior and an inverse relationship with CAS and bio- mass. We also investigated the possible relation between retention time and primary production. Experimental approaches to investigate neuromast re- generation of the axolotl lateral line. SPECK, R. R. and K. J. BALAK,* Department of Biology, Western Kentucky University, Bowling Green, KY 42101. The lateral line system of amphibians consists of dis- crete organs called neuromasts distributed in lines within the epidermis. The posterior lateral line runs along the dorsal midline of the tail. If the tail tip of an axolotl is amputated it will regenerate. The posterior lateral line will also regenerate about the same number of neuromasts as were removed with the tail tip amputation. Vital-dye la- 94 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1—2) belling experiments have shown that the regenerated neu- romasts are derived exclusively from cells of the last neu- romast left on the tail after tail tip amputation. Single-cell labeling experiments have implicated the supporting cells as the source of cells for the regenerated neuromasts. Par- tial tail tip and neuromast amputations were performed and the effect of these manipulations on neuromast re- generation examined. Three types of experiments were performed: (1) removal of the neuromasts without tail am- putation, (2) amputation of the dorsal tail tip and neuro- masts, but not the spinal cord, and (3) amputation of the ventral tail tip and spinal cord, but not the neuromasts. Glycoconjugate expression in normal and regenerating lateral line sensory epithelia of the axolotl salamander, Ambystoma mexicanum. MILLER, RICHMOND J. and K. J. BALAK,* Department of Biology, Western Kentucky University, Bowling Green, KY 42101. The sensory epithelia of the lateral line system of am- phibians and the inner ear of birds and mammals consists of two cell types: sensory hair cells and supporting cells. If the hair cells of mammals are destroyed, they are not replaced, resulting in permanent hearing impairment. This is not the case in amphibians and birds. If the hair cells of the cochlea are destroyed in young chicks they are replaced by regeneration. Experiments with the amphib- ian lateral line have shown that the regenerated hair cells are derived from progeny of supporting cell divisions. The hair cells of lateral line neuromasts specifically accumulate a fluorescent dye abbreviated DASPEI. When exposed to blue light the DASPEI labeled hair cells are killed by pho- totoxocity. These hair-cell-depleted neuromasts will regen- erate new hair cells in a process similar to that of a chick cochlea. Lectins are plant proteins that bind to monosac- charides and short polysaccharide chains. Biotinylated lec- tins were used to compare glycoconjugates expressed in normal intact neuromasts and neuromasts depleted of hair cells. SCIENCE EDUCATION A model classroom curriculum integrating core con- cepts of biomedical research/societal values to achieve KERA learner outcomes in secondary science classrooms. STEPHANIE WYATT, Calloway County High School, Murray, KY 42071, GREGORY J. POPKEN, University of North Carolina, Chapel Hill, NC 27599, and CLOYD J. BUMGARDNER, Calloway County High School, Mur- ray, KY 42071. A basic tenet of school reform mandated by the Ken- tucky Education Reform Act of 1990 is integration of knowledge. This goal necessitates the development of dis- trict, building, and classroom-level curriculum. In this model classroom curriculum, high school students com- pleted a written questionnaire designed to elicit their opinions of how biomedical research utilizing animal stud- ies impacted their everyday lives. The students then re- ceived instruction on the importance of this research to themselves and society based on opinions of researchers who are active in the field. The information was presented in audio-visual, lecture, and cooperative learning formats. The depth and breadth of instruction were based on pre- vailing societal concerns and a teacher assessment of stu- dent opinions noted on the questionnaire. Student com- pletion of the questionnaire prior to instruction provided the instructor with a baseline of information from which to proceed when teaching. Completion of the same ques- tionnaire after instruction indicated a broadening of stu- dent perceptions of the role biomedical research plays in our society's development. Are Kentucky students sleeping enough? JOHN G. SHIBER, Division of Biological Sciences & Related Tech- nologies, Prestonsburg Community College, Prestonsburg, KY 41653. Over 3,000 western Kentucky students (K—College) were surveyed about their sleep habits. The majority of K-—5 graders said they sleep 9-10 hr per night. Most 6-8 graders get 8—9 hr, but nearly a quarter of them get fewer than 8. Ninth to 12 graders generally get 7 or fewer hr (21% get 6 or less), and, although 99% of college students (18 yr->40) said adults should get 7-8 hr of sleep per night, only 40% of them get that amount. Except for the 4% who get more, the majority get 6 or fewer hr of sleep per night. Not surprisingly, 45-55% of all students sur- veyed have difficulty sleeping at night. Also, the older the student, the less rested he/she seems to feel upon waking, i.e., 29% of K-5, 39% of 6-8, 55% of 9-12, and 59% of college students do not feel well rested in the morning. Naps are a good idea, according to 71% of college stu- dents, but only 17% take them regularly. 41% of 9-12 graders take naps on days off, and 24% of them sleep in school, as opposed to only 8% of K-8 graders who do. After-school activities occupy 62% of all K-12 students surveyed. K—5 graders are out 1-2 evenings per week, usually getting home by 9 pM, and often before 7 pm; 6— 8 graders are involved 4-5 nights and also get home be- fore 9 pm; and 9-12 graders get home from their 3-5 nights of activities by 10 pm, except for 19% who get home later. To top it off, 66% of K-12 students watch TV until bedtime! About 14% watch only 1 hr or less, but 43% watch 2-3 hr and 44% watch 4 or more hr each night. An alarming statistic now exists that drowsy drivers cause more fatalities per vehicle accident than drunken drivers. In this survey, 56% of college students get sleepy while driving and most of them keep on driving anyway, doing various things to remain awake! Last, according to this survey, 18% of K—5, 24% of 6-8, and 45% of 9-12 graders do not eat any breakfast. This is very disturbing. How effective can educational reform in Kentucky (or any- where else) be when children not only go to school poorly rested, but also without having eaten? Surely some of our reform efforts should be focused on instilling the impor- tance and rewards of good eating/sleeping habits, the most basic of biological disciplines, in students and parents alike. After all, “A healthy mind is in healthy body!” PROGRAM, ANNUAL MEETING 95 Enhancing student perceptions of science and achieving Kentucky Education Reform Act learner outcomes through a creative science classroom curriculum. STE- PHANIE L. WYATT,* Calloway County High School, and CLOYD J. BUMGARDNER,* Calloway County Middle School, Murray, KY 42071. Student opinions of science may often be determined by the extent to which they perceive science impacting their daily lives. Many factors, including but not limited to overwhelming curricular and social concerns, time con- straints, and disciplinary requirements, may limit the ex- tent to which students retain factual knowledge presented in a lecture format. A model classroom curriculum em- ploying real life experiences as an instructural foundation was developed to address student perceptions of biotech- nology and microbiology. Student opinions of their own level of awareness of biotechnological issues were deter- mined by using a short questionnaire before and after in- struction. In addition to enhancing student opinions and sensitivity to these areas, the curriculum was designed to address several of the learner goals specified by the Ken- tucky Education Reform Act of 1990. Success under the goal-centered focus of the Reform Act mandates a teach- ing methodology dissimilar to the more traditional lecture- oriented approaches previously used in many classrooms. ZOOLOGY AND ENTOMOLOGY Equine protozoal myeloencephalitis: a review. S. STAMPER* and D. E. GRANSTROM, Department of Veterinary Science, Gluck Equine Research Center, Uni- versity of Kentucky, Lexington, KY 40546-0099. Equine protozoal myeloencephalitis (EPM) was first described in 1964 as segmental myelitis. In the mid-1970s, organisms were found to be associated with the lesions. S. W. Davis, a post-doc with J. P. Dubey, was the first to culture Sarcocystis neurona; Dubey later named the or- ganism S. newrona. The disease occurs only in the western hemisphere, with cases reported from North, South, and Central America. All breeds of horses are affected. The life cycle of S. newrona is not known; however, it should be similar to other members of the genus Sarcocystis. The genus typically has a predator/prey or scavenger/carrion life cycle. Horses appear to be aberrant, dead-end hosts with no sarcocysts being found. Therefore, transmission to the definitive host is prevented. Many species have been suggested as the definitive host. However, only skunks have shown the presence of antibodies to S. neu- rona-specific proteins, but they, like horses, may have been exposed inadvertently. The response of horses to the parasite probably depends on infective dose, immune competency, and stress. Relapses can occur after treat- ment. Such relapses represent a latent stage or a small focus of infection or reinfection. Immunodiagnosis is pos- sible with Western Blot, using cerebrospinal fluid (CSF) or serum, Negative serum is more conclusive than posi- tive. Positive serum demonstrates exposure, while a posi- tive CSF indicates the parasite has crossed the blood/brain barrier and has stimulated a local immune response. Trans. Ky. Acad. Sci., 56(1-2), 1995, 96-98 DISTINGUISHED SCIENTIST AND OUTSTANDING TEACHER AWARDS, 1994 OUTSTANDING SECONDARY SCIENCE TEACHER Beverly Lynn White—Instructor, North Lau- rel High School, London, Kentucky 40741 Native of Charleston, West Virginia Education: Marshall University—B.A. Biology and Physical Science; Marshall University— M.S. Biology; Post graduate courses at UK, EKU, Marshall, Gna College Lynn White has taught at Paul Blazer H.S. in Ashland, KY; at Sue Bennett College from 1972-1989; at South Laurel H.S. 1989-1992; and at North Laurel H.S. since 1992. She teaches Anatomy & Physiology; AP Bi- ology; Biology; Introduction to Physics and Chemistry; and Ecology. She has attended numerous workshops and conferences for teachers: Anatomy & Physi- ology for H.S. teachers—UK 1990; Rainforest Ecology workshop—Lima, Peru, 1992; Wild- life Management workshop—Cumberland Falls St. Pk, UK; DNA Science workshop— UK, 1994. She has attended the Kentucky Science Teacher Association Conferences and the Na- tional Science Teacher Association Confer- ences. She maintains many memberships in sci- entific organizations and has been a member of KAS since 1977. Lynn has received the Outstanding Appa- lachian Teaching Award from the Appalachian Scholars Program in 1991. She was recognized for allenic in teaching by the Covenen. S Scholars Program. She was voted the Teacher of the Year at North Laurel H.S. for 1991— 1992. Lynn has been involved with the state’s Wa- ter Watch Program at Levi Jackson State Park with her Ecology class. She received a grant to develop an outdoor classroom at North Laurel H.S. and this has become an interdis- ciplinary school-wide project. She serves as a volunteer guide for wild- flower tours of the Daniel Boone National Forest giving tours to campers and visitors. She gives talks about her experiences esp. Rainforest Ecology to schools and civic organ- izations. 96 As a teacher she motivates students and al- lows them to develop in the curriculum area of science. She excites children. She cares for students and stays after school to give extra help. Students stated that “she is never boring, and gives practical application to what she teaches.” She stresses “hands-on.” Lynn White has a genuine interest in her students. She is the kind of teacher who is “often discussed and admired at our family dinner table.” OUTSTANDING COLLEGE/UNIVERSITY SCIENCE TEACHER Dr. Barbara A. Ramey—Professor of Biologi- cal Sciences, Eastern Kentucky University Native of Freeport, Illinois Education: Cornell College, Mt. Vernon, [A— B.A. Biology (1968); Miami University, Ox- ford, OH—M.S. Zoology (1971); University of Kentucky—Ph.D. Biology (1982) Barbara Ramey is a versatile teacher who is prepared, highly competent, and very effec- tive. She expends many hours counseling stu- dents. She is at home in a freshman Zoology course, an upper level Cellular Biology class, or in a graduate embryology or histology course. She is highly involved with the Premedical Sciences program. She is active in a number of professional organizations, including the Kentucky Acade- my of Science, where she has served as mem- bership chairperson. Barbara is her department’s Coordinator of Graduate Programs. She maintains an active research agenda in- volving the effects of acid pH on embryonic and juvenile stages in fish; teratogenic effects of aquatic pollutants on embryos of freshwater fish and amphibians; and biomonitoring a con- structed wetland site. Her research has re- ceived funding from the U.S. Dept. of Interior and the U.S. Geological Survey via the KY Wa- ter Resources Research Institute. She has presented many papers at profes- sional meetings. She also has continued to AWARDS 97 publish her research in regional and national journals. Additionally, Barbara has been a member of numerous committees at Eastern Kentucky University. In 1989, she received the EKU National Alumni Association Award for Teaching Ex- cellence. That is a student nominated and stu- dent recommended award. As a teacher, Barbara has the natural ability to make difficult concepts easier for her stu- dents to grasp. She loves teaching and stu- dents realize her dedication and compassion for them. She is sincerely concerned about her students and schedules review sessions for her classes. She even comes in on weekends to help her classes. She is an excellent counselor for students who have expressed an interest in science. She involves students in her research. She brings innovation to her classes. Her ex- ams are challenging and require a sound un- derstanding and critical analysis of the mate- rial. She is friendly and approachable. S$ She communicates with her classes. Dr. Ramey has chosen to make teaching her top priority. DISTINGUISHED SCIENTIST AWARD Dr. Donald T. Frazier—Professor of Biomed- ical Engineering, Department of Physiology and Biophysics, University of Kentucky Col- lege of Medicine Native of Floyd County, Kentucky Education: B.S. University of Kentucky (1958): M.S. University of Kentucky (1960): Ph.D. University of Kentucky (1964) After receiving his Ph.D., Donald Frazier taught at the University of New Mexico School of Medicine where he remained until 1969 when he returned to the UK College of Med- icine. He has remained there in the Depart- ment of Physiology since, first as Associate Professor and since 1974 as Professor. From 1980-1992, he served as Chair of the Department of Physiology. He has been rec- ognized as a creative scientist and outstanding teacher. Professor Frazier has distinguished himself in three areas: research and scholarly activity, education, and out-reach educational pro- grams. Don ranks among the university’s most dis- tinguished scientists, whose research has been deloummle dged nationally and internationally. He has maintained a well-funded research lab- oratory by the National Institutes of Health that focuses on the neural regulation of breathing, for the last 20 years. In pddition to Don's highly effective class- room teaching of medical, doctoral, and un- dergraduate students, he has utilized his re- search laboratory to train postdoctoral fellows in the intricacies of neurobiological research. He has also made contributions on the nation- al level by being the Director of the Summer Fellowship Program at the renowned Marine Biological Laboratories in Woods Hole, Mass. Moat importantly, Don has provided lead- ership within the university to establish sci- ence out-reach programs for under- -represent- ed students including high school and under- graduate minority students, to bring them to ihe campus to provide an opportunity to ob- tain first hand experience in biomedical, health-related and scientific research. It is im- portant to recognize that he has competed successfully on the national level to obtain ex- tramural funding from the National Institutes of Health for this minority educational pro- gram. He is a person who remembers his Appa- lachian roots and has worked hard to get the school systems involved in higher aduciton He has maintained a long “Hist of member- ships in professional societies at the state, re- gional, and national level. He has directed doctoral research and his list of publications is extensive. He has accepted his responsibilities very se- riously. He was named to the Commission on Human Resources and Social Change of the National Association of State Universities and Land Grant Colleges. INDUSTRIAL SCIENTIST OF THE YEAR 1994 Dr. Fred L. Tungate—Manager of Research and Development, United Catalysts, Inc., Louisville, KY Native of Indiana Education: Indiana University—A.B. Chemis- try (1972); University of Tennessee—Ph.D. Chemistry, specializing i in Inorganic Chem- istry (1978) Dr. Tungate has been an industrial re- 98 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(1-2) searcher since receiving his doctorate. Since 1977, he has worked exclusively in the area of catalyst and adsorbent research, development, and commercialization. Dr. Tungate holds six patents and has au- thored a number of publications. He is exceptionally effective in interacting with technical personnel of client companies. His contributions in research and in working with clients has contributed significantly to the major expansion of existing product lines and the construction of new plant facilities. His work has broad applications including the edible oil hydrogenation to produce salad oils, frying oils, confectionery products. He in- vented the G-95 catalyst which has been the industry standard for 14 years. Additionally, he has worked with the highly advanced technol- ogy of zeolites. Fred is a member of a number of profes- sional organizations. He has been a part-time lecturer at the Uni- versity of Louisville, teaching a graduate course in Zeolite Chemistry. He has been a lecturer at the Mexican Institution of Petro- leum International. He is an excellent example of an Industrial Scientist at work, keeping abreast of the latest technology in his chosen field of catalysis, while also participating in programs to pro- mote scientific interests in society and _his community. He is now the Director-at-Large to The Ca- talysis Society and has served in numerous ca- pacities of the Tri-State Catalyst Club. Dr. Tungate’s career could be a model of what an industrial scientist should be. Trans. Ky. Acad. Sci., 56(1-2), 1995, 99 NEWS The annual meeting of the Kentucky Acad- emy of science for 1995 will be a joint affair with the Tennessee Academy of Science, 16, 17, 18 November. All members should make an effort to attend this forum. The 1996 meeting will be at Kentucky State University in Frankfort, and the 1997 meeting will be at Morehead State University. Sy) Instructions for Contributors s a Original papers based on research in any field of science will be considered for publication ‘in the Transactions. Also, as the official publication of the Academy, news and announcements _ of interest to the membership will be included as received. _ Manuscripts may be submitted at any time to the Editor. Each manuscript will be reviewed _ by one or more persons prior to its acceptance for publication, and once accepted, an attempt will be made to publish papers in the order of acceptance. Manuscripts should be typed double _ spaced throughout on good quality white paper 8% x 11 inches. NOTE: For format of feature _ articles and notes see Volume 43(3-4) 1982. The original and one copy should be sent to the _ Editor and the author should retain a copy for use in correcting proof. Metric and Celsius units _ shall be used for all measurements. The basic pattern of presentation will be consistent for all _ manuscripts. 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The body of the manuscript should include the following sections: Introduction, Materials and Methods, Results, Discussion, Summary, Acknowledgments, and Literature Cited. All tables and figures, as well as all literature cited, must be referred to in the text. 4. All references in the Literature Cited must be typewritten, double spaced, and should provide complete information on the material referred to. See Volume 43(3-4) 1982 for style. _ 5. For style of abstract preparation for papers presented at annual meetings, see Volume 43(3- » 4) 1982. and set on a separate page. The heading of the table should be informative of its contents. Each figure should be reproduced as a glossy print either 5 x 7 or 8 Xx 10 inches. Line draw- _ ings in India ink on white paper are acceptable, but should be no larger than 8% X 11 inches. Photographs should have good contrast so they can be reproduced satisfactorily. All figures _ should be numbered in Arabic numerals and should be accompanied by an appropriate legend. ‘It is strongly suggested that all contributors follow the guidelines of Allen’s (1977) “‘Steps Toward Better Scientific Illustrations’ published by the Allen Press, Inc., Lawrence, Kansas 66044. ~ _ The author is responsible for correcting galley proofs. He is also responsible for checking all literature cited to make certain that each article or book is cited correctly. Extensive alterations on the galley proofs are expensive and costs will be borne by the author. Reprints are to be ordered when the galley proofs are returned by the Editor. CONTENTS Changes in vegetation and bobwhite quail and eastern cottontail rabbit use in a converted fescue field. Jeffrey D. Sole ............c0ccccccccecceeoees 1 Human sex ratio and factors influencing family size in Hunan, China. Elmer Gray, Valina K. Hurt, and Ju Y. Wu ...............0cceecceeseceeeecees 8 Decomposition of nerve gas and mustard gas analogs using nicotine and ultrasound. John L. Meisenheimer, Lawrence R. Miller, and William DS Sohal 2 oe OO es Ee AU, SANS Sy a nv a 15 Oocyte staging in paddlefish, Polyodon spathula. William L. Shelton and: Steven, DOMUMS eo oie das Seca oe aece abane tides Sera eee eine ree 22 Egg and larval development of the striped fantail darter, Etheostoma flabellare lineolatum (Agassiz), and duskytail darter, E. percnurum Jenkins, with comments on the Etheostoma flabellare species group. Thomas P. Simon and Steven R. Layman .............0.....ecceeecesseccceees 28 Aspects of the dragonfly and damselfly (Odanata) community of Buck | Creek, Pulaski County, Kentucky. Randall G. Payne and Guenter A. SGRUSEOR S22) SoBe ia reas oie di wade ens ek cod Senate ale easter Gdlaignmemetela GeeaiGe wtaek Sara ama 41 Serological evidence for Borrelia sp. in Peromyscus leucopus from Western Kentucky. Zixing Wang, Leon Duobinis-Gray, Stephen White, Ed Snoddy, and James Stuart oicc2cicc was assesses Wovsestese posbenscmnereeee neces 47 Pennsylvania sharks from Kentucky. James X. Corgan ..................++ 54 NOTE New records of slender madtom, Noturus exilis Nelson, South Fork Licking River, Pendleton County, Kentucky. Lewis E. Kornman ........ 57 ACADEMY: AFFAIRS 0c 5 odes cee cea cuuscceianen ccteemepocauiectcum@bucacknine doa deusdyceen ea mme 58 PROGRAM, ANNUAL MEETING ji oucco ooo oi sec et os eat ccavien tuonsacsedeasesecensaeuame 62 ABSTRACTS OF SOME PAPERS PRESENTED AT THE ANNUAL MEETING, ec ue oa e baw nde sida ube Geman svenmngh Mee aR MNase Shee eal UUM MI ZL at Us Oty a 80 DOA rec lecskec Woes ataoa san VackeW ned teles OU CURCUMIN a a a ee 96 TRANSACTIONS DF THE KENTUCKY ACADEMY OF SCIENCE PEA SGN Volume 56 + 18 1065 Numbers 3-4 i " | September 1995 Bi... : Official Publication of the Academy The Kentucky Academy of Science Founded 8 May 1914 GoverRNiING BOARD FoR 1995 Executive COMMITTEE President: Robert Creek, Department of Biological, Eastern Kentucky University, Richmond, KY 40475 President Elect: William S. Bryant, Department of Biology, Thomas More College, Crestview Hills, KY 41017 Vice President: Marcus T. McEllistrem, Department of Physics, University of Kentucky, Lexington, KY 40506-0055 Past President: Larry P. Elliott, Department of Biology, Western Kentucky University, Bowling Green, KY 42101-3575 Secretary: Peter X. Armendarez, Department of Chemistry and Physics, Brescia College, Owensboro, KY 42301 Treasurer: eee Ef Carter, Wood Hudson Cancer Research Laboratory, 931 Isabella Street, Newport, KY 107 Executive Secretary (ex officio): J. G. Rodriquez, Department of Entomology, University of Kentucky, Lexington, KY 40546-0091 Editor, TRANSACTIONS (ex officio): Branley A. Branson, Department of Biological Sciences, Eastern Kentucky University, Richmond, KY 40475 Editor, NEWSLETTER (ex officio): Vincent DiNoto, Natural Science Division, Jefferson Community Col lege, Louisville, KY 40272 MEMBERS, GOVERNING BOARD Blaine R. Ferrell 1995 Wimberly C. Royster 1997 Valena Hurt 1995 James Hopgood 1998 Patricia K. Doolin 1996 Robert H. Wombles 1998 David E. Hogan 1996 AAAS/NAAS Representative Gerald L. DeMoss 1997 J. G. Rodriguez 1998 Chairperson, KJAS Valgene L. Dunham 1995 COMMITTEE ON PUBLICATIONS Editor and Branley A. Branson, Department of Biological Sciences, Eastern Kentucky University, Chairman: Richmond 40475 Associate Editor: Thomas K. Green, Department of Chemistry, Western Kentucky University, Bowling Green 42101 Abstract Editor: Robert F. C. Naezi, Department of Biological Sciences, Northern Kentucky University, Highland Heights 41076 Index Editor: Varley Weideman, Department of Biology, University of Louisville, Louisville 40292 Editorial Board: Vincent DiNoto, Natural Science Division, Jefferson Community College, Louisville 40201 Jerry N. Baskins, School of Biological Sciences, University of Kentucky, Lexington 40506 John P. Harley, Department of Biological Sciences, Eastern Kentucky University, Richmond 40475 Robert Creek, Department of Biological Sciences, Eastern Kentucky University, Richmond 40475 Larry P. Elliott, Department of Biology, Western Kentucky University, Bowling Green 42101 John Thioret, Department of Biology, NounoE: Kentucky University, Highland Heights 41076 All manuscripts and correspondence concerning manuscripts should be addressed to the Editor. Authors must be members of the Academy. The TRANSACTIONS are indexed in the Science Citation Index. Coden TKASAT. ISSN No. 0023-0081. Membership in the Academy is open to interested persons upon nomination, payment 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. Annual dues are $25.00 for Active Members; $15.00 for Student Members; $35.00 for Family; $350.00 for Life Mem- bers. Subscription rates for nonmembers are: domestic, $45.00; foreign $50.00; back issues are $30.00 per volume. The TRANSACTIONS are issued semiannually in March and September. Four numbers comprise a volume. Correspondence concerning memberships or subscriptions should be addressed to the Secretary. Exchanges and corre- spondence relating to exchanges should be addressed to the Librarian, University of Louisville, Louisville, Kentucky 40292, the exchange agent for the Academy. This paper meets the requirements of ANSI/NISO Z39.48-1992 (Permanence of Paper). ee INDUSTRIAL AFFILIATES ASSOCIATE PATRON ASHLAND OIL, INC. MEMBER CoORHART REFRACTORIES CORP. MPD, Inc. UNITED CATALYsT, INC. ASSOCIATE MEMBER 3M Tare MANUFACTURING DIVISION, CYNTHIANA PLANT ALL-RITE PEst CONTROL HorEcHsT CELANESE CORP.-ENGINEERING PLASTICS DIVISON KENTUCKY SCIENCE & TECHNOLOGY COUNCIL, INC. WESTVACO INSTITUTIONAL AFFILIATES FELLOW University of Kentucky SUSTAINING MEMBER Eastern Kentucky University Morehead State University Murray State University Northern Kentucky University University of Louisville Western Kentucky University MEMBER Campbellsville College Cumberland College Kentucky State University Paducah Community College Southeast Community College ASSOCIATE MEMBER Ashland Community College Berea College Brescia College Centre College Georgetown College Hazard Community College Jefferson Community College Kentucky Wesleyan College Lees College Maysville Community College Midway College Owensboro Community College Somerset Community College Spalding University Thomas More College Transylvania University f mC Nae ee ert Mo) ay 3 his Hf * ort Cl ee - soph Linen TRANSACTIONS of the KENTUCKY ACADEMY of SCIENCE September 1995 Volume 56 Numbers 3-4 Trans. Ky. Acad. Sci., 56(3-4), 1995, 101-103 Daphnia lumholtzi: Appearance and Likely Impacts of an Exotic Cladoceran in the Ohio River JEFFREY D. JACK AND JAMES H. THORP Large River Program, Department of Biology, University of Louisville, Louisville, Kentucky 40292 ABSTRACT Exotic species can have a profound impact on the ecosystems they invade. Over the period 1992-1994, the exotic cladoceran Daphnia lumholtzi has become increasingly common in the Ohio River. This cladoceran has prominent helmet and caudal spines which may protect it against invertebrate and some vertebrate predators. This well-defended cladoceran may eventually dominate the Ohio River plankton during the warm months of the year, potentially altering the composition of the potamoplankton assemblage and the trophic processes within the river. INTRODUCTION The Ohio River and its tributaries have been invaded by several exotic species in re- cent history. Species with obvious direct eco- nomic impacts such the zebra mussel (Dreis- sena polymorpha) have been the object of in- tense research attention, but there are new ar- rivals in the plankton community of the Ohio that may have less direct but equally serious effects. ‘Among these is the exotic water flea, Daphnia lumholtzi Sars [Crustacea: Branchi- poda] which may have been accidentally intro- duced from its native habitat in Africa, Asia, or Australia to the southern United States in association with either fish-stocking efforts or the release of exotic aquarium fish into the environment (1). It is distinguished from other cladocerans in the Ohio River by its relatively large size (adults average, about 1.1 mm long, [2]) and the prominent spines on the head (helmet) and caudal regions. The plankton community in the McAlpine pool of the Ohio has been dominated for the past 4 years by small cladocera, such as Bosmina longirostris, which have no obvious spines (3). The presence of spines in cladocera is often considered an anti-predator mechanism to re- 101 duce success by vertebrate and invertebrate predators in capturing and ingesting these or- ganisms. The recent invasion of Lake Michi- gan by the predatory cladoceran Bythotrephes cederstroemii shows the impact that such well- defended zooplankton can have. This northern cladoceran also has a prominent caudal spine, which makes capture and ingestion by small fish such as juvenile yellow perch (Perca fla- vescens) difficult. Apparently these fish learn to avoid attacking B. cederstroemii (4). Such aversive behavior by the fish may prevent them from wasting pursuit and handling en- ergy attempting tes capture B. cederstroemii, Buk it also allows this cladoceran to compete with these small fish for other, less well-de- fended zooplankton prey. To determine the seasonal occurrence, den- sities and distribution of D. lumholtzi at sites in the lower McAlpine and upper Cannelton pools of the Ohio River, we collected numer- ous plankton samples from ae through fall of 1992-1994. If the densities of D. honiholta i are increasing in the Ohio, it is likely this cla- doceran will fe e a negative impact on the na- tive zooplankton and on the fish that rely on these zooplankton for food. METHODS We took plankton samples every 10 days from April to November at 2 sites in the Ohio River in 1992-1994. Two shore (within about 10 m of the shoreline) and one midchannel sample were taken at Cox’s Park (Ohio River Mile [ORM] 600) near Louisville, KY and the Gallagher Power Station in New Albany, IN (ORM 610). Fifty liter samples were retrieved using a battery-operated water pump (rate ~ 26 liters min.') from 1.0 m depths through a 63 wm-mesh plankton net. The samples were concentrated through a dolphin bucket (also with 63 jm mesh) at the cod-end, placed in 75 ml N algene screw-top containers trans- ported back to the laboratory on ice. The sam- ples were counted and identified within 4 hours of their collection. The zooplankton samples were enumerated at 45X using a Ni- kon SMZ-10 or a Nikon SMZU stereomicro- scope. Daphnia lumholtzi was identified using the drawings of Havel and Herbert (1). In 1994, various physical data were also taken in the river at the Cox’s Park site. Tem- perature and conductivity data at the river sur- face were collected using a YSI Model 57 ox- ygen meter. Water samples (75 ml) were taken from the river surface, stored in acid-washed plastic containers and later analyzed for tur- bidity in nephelometric turbidity units (NTUs) using a Hach model 2100P turbidity meter. RESULTS AND DISCUSSION We collected no physical data from the Ohio with our plankton samples during 1992— 1993, but in 1994 the conditions in the river varied seasonally. Turbidity was highest in April (mean ~ 8.2 NTUs) and dropped through the year until the final sample in No- vember (~3.9 NTUs). Temperature varied from 17°C (April) to a high of 28°C in late August. Temperatures had dropped to about 15°C in November when the sampling ended. Conductivity did not vary much through the sampling period, remaining between 320 and 490 wmohs. Daphnia lumholtzi was present in low den- sities in the summers (June through August) of 1992 and 1993 (5), but the occurrence and densities were too sporadic and low to distin- guish any meaningful trends. In 1994, D. lum- holtzi appeared suddenly in our samples in TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) Number Daphnia lumholtzi_L-1 = > o => a iS = a xt 2 [s) o Pees fp) Zz Month Sampled Fic. 1. Abundance of Daphnia lumholtzi in the Ohio River in 1994. Means are the result of at least three sam- ples within the month; error bars indicate standard devi- ations. See text for comparison to densities in 1992-1993. late July (see Fig. 1) at significant densities in the river. Through most of August, D. lum- holtzi was a significant component (about >50% numerically) in our samples of the crustacean zooplankton in the Ohio River. Its population levels dropped soon afterward to undetectable levels in late August, although it reappeared briefly at lower densities in Octo- ber (see Fig. 1). The appearance of this exotic zooplankter is cause for concern both in terms of the trophic relationships in the Ohio River plankton com- munity and its resultant impact on fisheries. Since its first confirmed report in 1991, D. lumholtzi has been found in reservoirs throughout much of the Southeast and Mid- west regions of the United States (1); its rapid dispersal would indicate it could potentially colonize much of the Ohio River within the next few years. While D. lumholtzi may not prey directly on other cladocerans as B. cederstroemii does, it is still likely to have a negative impact on lotic and lentic systems in this country. The pres- ence of the spines on its helmet and caudal regions may provide some protection against invertebrate and small vertebrate predators. DAPHNIA LUMHOLTZI IN KENTUCKY—Jack and Thorp Such structures have been shown to be effec- tive against invertebrate predators in rotifers (6) and daphniids (7), as well as against fish (4). As D. lumholtzi continues its colonization of the Ohio River and tributaries, its defenses may result in it becoming a dominant member of the zooplankton during the warmer por- tions of the year. It seems likely that inverte- brate predators such as the phantom midge larva, Chaoborus (especially early instars), may not be able to feed efficiently on this well- protected cladoceran, which may result in more predation pressure on co-occurring but less well-defended cladocera such as Bosmina. Increased feeding on Bosmina would further drive the plankton community toward domi- nance by D. lumholtzi, although Bosmina may be able to compensate for its losses through increased reproduction. We will be performing both in situ and large tank experiments this spring and summer, in- vestigating the impact of this new cladoceran on the pelagic communities in the river. Our experiments will enable us to understand bet- ter the probable impact of this new member of the Ohio River plankton community on tro- phic processes in the Ohio River. SUMMARY In systems dominated by D. lumholtzi, a new population bottleneck for large inverte- brates and small vertebrates may emerge. Densities of small zooplanktivorous fish and 103 the macroinvertebrate predators such as Chaoborus, which can be important food for the larger fish in a community, may drop due to poor success with the new dominant cla- doceran. Decreases in their densities would probably have an impact on higher trophic lev- els, including the larger fish popular with sport fishermen. ACKNOWLEDGMENTS The sampling expertise of Andy Casper, Kim Greenwood and Richie Kessler is grate- fully acknowledged. This work was supported in part by grants from PSI Energy and the U.S. Environmental Protection Agency. LITERATURE CITED 1. Havel, J. E. and P. D. N. Herbert. 1993. Daphnia lumholtzi in North America: another exotic zooplankter. Limnol. Oceanogr. 38:1823-1827, 2. Thorp, J. H., A. R. Black, K. H. Haag, and J. D. Wehr. 1994. Zooplankton assemblages in the Ohio river: seasonal, tributary and navigation dam effects. Canadian J. Fisheries and Aquatic Sci. 51:1634—1643. 3. Jack, J. D. unpublished data. 4. Barnhisel, D. R. 1991. The caudal appendage of the cladoceran Bythotrephes cederstroemii as a defense against young fish. J. Plankton Res. 13:529-537. 5. Black, A. R. personal communication. 6. Stemberger, R. S. and J. J. Gilbert. 1987. Multiple species induction of morphological defenses in the rotifer Keratella testudo. Ecology 68:370-378. 7. Havel, J. E. and S. A. Dodson. 1984. Chaoborus predation on typical and spined forms of Daphnia pulex: behavioral observations. Limnol. Oceanogr. 29:487-494. Trans. Ky. Acad. Sci., 56(3-4), 1995, 104-108 Comparing Epizootic Systems Using Spectral Analysis and Autoregression: A Case Study on Tetranychus—Neozygites Mycosis F. X. Susito, G. L. Norprin, G. C. BRown, AND R. HasiBuAN Department of Entomology, University of Kentucky, Lexington, Kentucky 40546 ABSTRACT Host-pathogen interactions at the population level were studied using the twospotted spider mite, Tetran- ychus urticae Koch, and the pathogenic fungus, Neozygites floridana Weiser and Muma, as a model system. The generation time (egg to adult) of the twospotted spider mite is approximately 10 days while the gen- eration time of N. floridana (host penetration to release of spores) is approximately 4 days when both are reared at 21°C. Three different systems were studied in relation to epizootic events. In System 1, leaf disks were infested with 250 eggs, 125 immatures, 5 male adults, and 20 female adults. In System 2, 25% of the immatures were added every other day and in System 3, 25% of the immatures were removed every other day. Our objective was to change the rate of increase of the host population. These systems were observed for 200 days. Spectral analyses showed that regulation of the host existed under 3 conditions. First, the pathogen could regulate the host in an epizootic system in which no hosts were removed but in which pathogens were periodically added. Regulation also occurred in systems in which hosts were added or removed, as long as a critical number of pathogen units (mummies) was maintained. Finally, a single intro- duction of pathogens could regulate the host in systems in which a portion of hosts was periodically removed. Autoregression and cross-correlation analyses of this system indicated that the number of mites that became infected can almost always be predicted by the number of eggs, but it cannot be predicted by the number of immatures. Moreover, densities of infected hosts could be forecast from a range of time lags of 2-16 days except for a time lag of 8 or 10 days. INTRODUCTION An epizootic system consists of 3 basic com- ponents: host, pathogen, and environment (1, 2, 3). Under a given set of environmental con- ditions, host-pathogen interactions are affect- ed by host density, susceptibility, behavior, in- terspecific characteristics (4), pathogen densi- ty, infectivity, latency, survival, spatial distri- bution (5) and transmission characteristics (6). Simple models have long been used to study the elementary dynamics of such systems (7, 8, 9). These authors differentiate the host pop- ulation into 2 distinct subpopulations of sus- ceptible and infected hosts which were treated separately. Similar approaches have also been proposed by Anderson and May (10, 11, 12) who treated the host component as a compos- ite of the 2 subpopulations. These studies have resulted in establishment of the threshold density concept with various epizootiological implications (13, 14) and unification of ecolog- ical (predator-prey) and epizootiological (pathogen-host) models (15). Epizootic systems are often periodic which means that they can be analyzed by identifying the dominant cycles (16). One way to identify those cycles is to use spectral analysis. This analysis has been successfully used in identi- fying periodicities in human epidemics (17). When the spectral density (variance of evenly spaced data points) is plotted against its fre- quency, power spectra peak in the dominant cycle(s) of the system. The biological impor- tance of the cycle(s) is identified by correlating it with the biological characteristics of the sys- tem of interest. In this study, we evaluated 2 methods, spec- tral analysis and autoregression, to analyze ep- izootic systems using the twospotted spider mite, Tetranychus urticae Koch as infected by the pathogenic fungus, Neozygites floridana Weiser and Muma, as an experimental model system. — MATERIALS AND METHODS Experiments were carried out in the Insect Pathology Laboratory, Department of Ento- mology, University of Kentucky, Lexington, KY, during January—April 1991. The epizootic model for this study consisted of the twospot- ted spider mite, Tetranychus urticae Koch, reared on bush bean plants (Taylor Strain) and the pathogenic fungus, Neozygites floridana Weiser and Muma, maintained in mummified mite cadavers. The original source of N. flor- 104 SPECTRAL ANALYSIS OF EPIZOOTIC idana was obtained as a gift from Dr. George G. Kennedy of North Carolina State Univer- sity, Raleigh, NC. Only adult mite cadavers were used as inoculum in this study. On those days when inoculation was to be performed, a separate population of cadavers was inspected and all cadavers removed. Four hours later. newly-killed cadavers were selected for use as inoculum as described by Brown and Hasi- buan (18). Therefore, age of inoculum (newly- formed cadavers) never exceeded 4 hours. Ca- davers always sporulated if humidity was at 100% RH. Experimental units were plastic trays (14 X 17 em) with wet cotton batting supporting 4 bush bean leaf disks (dia. 3 cm) connected to each other by 1 X 5 cm hardware cloth bridge spans. From an initial pool of 36 units, im- matures and adults were placed on them and were randomly assigned to 3 groups of epi- zootic systems, i.e., System | without additions or removals of host mites. In System 1, each leaf disk was infested with 250 eggs, 125 im- matures, 5 male adults, and 20 female adults. System 2 was similar except 25% of mite im- matures were added every other day whereas in System 3, 25% of mite immatures were re- moved every other day. Each system was fur- ther subdivided into three subsystems or treat- ments, i.e., control with mites only, treatment A with a single pathogen introduction of 5 mummies, and treatment B with repeated pathogen introductions of 1 mummy every other day. Observations on prevalence of my- cosis (number of infected hosts or new mum- mies) and the surviving mites (number of sus- ceptible hosts) were done every other day. Be- tween any 2 observations, all trays were kept in a growth chamber (Percival, Model 1-35 L) set at 21 + 1°C and 100% RH. These systems were run until all mites died of mycosis or 200 days which ever came first. The data series of susceptible hosts were documented as the number of eggs, imma- tures, adult males, and adult females. The se- ries of susceptibles and’ infecteds (cadavers) were smoothed using a five point polynomial method (19). Spectral analyses and autore- gression were then conducted on these smoothed data. 1. Spectral Analysis In this study, the SAS SPECTRA procedure (20) was used to generate spectral density SystEM—Susilo et al. 105 500 > ary rai B32 SE a an 0.00 0.39 0.79 1.18 1.57 196 23 275 3.14 Frequency (0 - pi) Fic. 1. Spectral density plots for total mites in System 1. System 1A (solid line) has a single obvious peak while system 1B (dotted line) has no peak. System 1C (dashed line) has two peaks that satisfy the criterion that the peak must be at least 10% higher than the preceeding and suc- ceeding spectral densities. The numbers on the graph cor- respond to the entries for total mites, System 1 A, B, and C in Table 1 plots in each epizootic system from the time series of susceptibles and infecteds. Some pre- dictions about the epizootic systems were then inferred. 2. Autoregression and Cross-Correlation Analysis This analysis sought to correlate a data point with its future value with the expectation that the value at some future time (At) could be predicted by the original data point. To do this, pairs of regressor-predicted variables to be analyzed were S(t) versus S(t + At), S(t) versus I(t + At), I(t) versus S(t + At), and I(t) versus I(t + At), where S is the number of susceptibles and I is the number of infecteds. The values of At used were 2, 4, 6, 8, 10, 12, 14, or 16 days. The upper hae of 16 days lag was chosen in this study because the total de velopmental period of ‘the mites is about 16 days (21). RESULTS AND DISCUSSION The criterion for identifying peaks in spec- tral density (which imply periodic fluctuations in these population systems) was that an in- crease of at least 10% in the spectral density was followed by a decrease by at least 10%. An example of the spectral density curves ob- tained from the spectral analyses is shown in Figure 1. This example, chosen because it dis- plays the array of observed responses and demonstrates the use of this criterion, corre- 106 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) sponds to the total mite curves for System 1, populations with no additions or removals of immatures. System 1A, with a single introduc- ie aa z tion of the pathogen, has a single obvious eos spectral peak at 40 d. System 1B, continuous a = a é a 5S I1Sa pathogen introduction, has no peaks; declining e ro) See throughout the frequency interval. System 1C, E § ae mites without the pathogen, has 2 peaks (using a z § a the 10% criterion), 1 of which is at nearly the 3 & Bo |° 2 Peat 5 (ego || S same frequency as System 1A and another of ue ee ele SSI which is at 22 days. £ “eb is cube The periodicities of all 9 systems are shown ° iatea in Table 1. Over half (52%) of the variables 1 Bs investigated had one periodicity while 29% ae elie a rei had none and the remaining 19% had more 2. 8 Bee] [oj See ie than one. Populations in System 2 (host ad- i 3 = ditions) had more periodicities than those in = & 2 © the other systems. System 1 had the largest Be > overall mean (32.8 + 2.1 d), System 3 hadthe Bp g © © = smallest (26.2 + 3.1 d) and System 2 was in- Ys = Senior ae. termediate with a mean (+ S.E.) periodicity 3 = iy x ms of 30.3 + 2.9 d. None of these differences 2 fe zs were significant. once - The mean periodicity for each population is a a" BP | oun also shown in Table 1. These means were 5 5 = % |2|< || 4a compared using all possible pairings in a two- Ss g Sale 18 tailed t-test (P = 0.05). In Systems 1 and 3, ¢ pez ¢ the mite population with no pathogen had sig- = Z 2 ¢ nificantly faster periods than the system with = & g tS is bs a single pathogen introduction. In contrast, = sy alla a te Gos these 2 population means were almost iden- 2h 55 ; tical in System 2. Examining these means in = a =) order of increasing population growth rate 2% 3 Eas (3—removal of immatures, 1—no manipula- § 2 ee & tion of immatures, 2—adding immatures), re- : Sf EM Leas Sse Fa veals a clear trend of increasing periodicity © es =e 5 with increasing growth rate on the populations = = & | with no pathogen. However, populations with g o ie I the pathogen did not show an obvious trend. Se See lS This trend is somewhat clearer if one ex- “= Bae 2 |< SBS amines the individual variables in order of in- = § > fas, creasing growth rate. This trend is observable ‘© » « for immatures (System C), total mites (Sys- 2 5 ae tems A and C), proportion of eggs (SystemC). 2° S By ks Susilo had predicted that the infected hoss 2258) |°|" ge! would show periodicities of 15-16 d with lon- 2% z ger periods possible. This prediction was = eS ES based, in part, on the spider mite’s longevity %%™ = 9 which, under the conditions used here, is 15-4 2 & 4 Galicia ee ene 17 d (21). The results in Table 1 confirm ths 88 °5] 4 5 o Sea & in Systems | and 2 but not in 3 and, even ates alee oe aS then, only in the cases where the pathogen was repeatedly introduced. No regular peri- 21 17 20; 17 17 18 23.33 + 4.99 ef 24 18 16 23.00 = 2.04 31.56 + 6.82 bede 21; 17 22 17 18 28.10 + 5.93 bde 53 ili, 53; 18 20 16 SLL ae ZLB) BLA ae bell bed 46 PAS MT Bop IU 48; 20; 18 bd 48 60 23 36.0 + 5.67 ab 40 38 37 39.0 + 0.51 24 23 25 Mean + S.E. 27.46 + 2.06 Sign. Diff. 39; 22 c Total SPECTRAL ANALYSIS OF EpizooTic SystEM—Susilo et al. 107 TABLE 2. Significance test for time lags for predicting susceptibles or infecteds. Variables Lags (days) Predicted Predictor 2 4 6 8 10 12 14 16 s(t + At) poe. S(t) gcc KK KK oat =. 23 — * kk * I(t + At) S()cce = ek 3k ek Kk ak + r Sit + At);... I(t) — — — — — — a * S(t ar At) imam Sb nam ue is ste = =~ * ae is I(t + At) a, Sear ANDRES I(t) a — — — — --- — a I(t + At) I(t) ek 2k * me = * x eK — Non-significant * Significant at P < 0.05. ** Significant at P < 0.01. odicity was observed in the populations sub- jected to a single introduction of the pathogen. The pathogen did persist in these populations, sometimes at high levels of incidence, there just wasn’t a regular periodic fluctuation. Brown (in press) has presented evidence that the interaction between the host and pathogen is chaotic and aperiodic. Brown (22) and Hasibuan (23) have both suggested that mite populations on the leaf disks used here tend to overdamp. This is why periodicities would tend to increase with in- creasing growth rates when growth rates are manipulated by increasing the population by a predefined percentage of those already pres- ent. However, when the pathogen is present, it serves to reduce the mite growth rate there- by lessening the overdamping effect. Conse- quently, in these populations, the pathogen would tend to cause the periodicities to de- crease as more immatures are added. It is this apparent conflict between the mite’s intrinsic dynamics and those of the mite-pathogen sys- tem that cause some of the periodicities to de- crease in System 2. For the autoregression analysis, numbers of mite eggs, immatures, and infected hosts in System 2B were used to represent a continu- um of susceptible and infected host classes tested because the prediction about host- pathogen regulation in that system was the re- sistant to perterbation. Results of autoregres- sion and cross-correlation analyses on the above three host-pathogen classes revealed that the number of infected hosts can almost always be predicted by the number of eggs (Table 2). Results indicate that the number of eggs can be predicted by the number of in- fected hosts using the time lags of 14 or 16 days. However, the number of infecteds can- not be used to predict the number of imma- tures, and vice versa. Moreover, a future value of infecteds can almost always be predicted by a previous value of infecteds, except when time lags of 8 or 10 days were used. The same exception was true for self-predicting the number of immatures. Self-prediction of the number of eggs was possible using time lags of 2, 4, 12, 14, and 16 days. CONCLUSION Spectral density analysis on 3 epizootic sys- tems of Tetranychus—Neozygites mycosis showed that pathogen cycles were maintained in systems where pathogens were repeatedly introduced. Host-pathogen regulation may oc- cur in epizootic systems when a portion of that host was routinely added or removed or in sys- tems with host additions or removals as long as the pathogen base-level was maintained. Autoregression analysis on susceptibles (mite eggs or immatures) and infecteds (mite mummies) demonstrated that the number of infecteds can almost always be predicted by the number of eggs. Future values of infect- eds, eggs, or immatures can be predicted by their corresponding previous values for a range of time lags of no more than the devel- opmental time of the host (16 days), except for the time lags of 8 or 10 days. ACKNOWLEDGMENT F.X.S. and R.H. thank USAID-WUAE pro- ject for providing financial support during their graduate studies in the U.S.A. This paper (94-7-123) is in connection with a project of the Kentucky Agricultural Experiment Station 108 and is published with the approval of the Di- rector. LITERATURE CITED 1. Steinhaus, E. A. 1949. Principles of insect pathol- ogy, Ist ed. McGraw-Hill Book Co. Inc., New York. 2. McNew, G. L. 1960. The nature, origin, and evo- lution of parasitism. Pp. 19-69. In J. G. Horsfall and A. E. Diamond (eds.) Plant pathology: an advanced treatise. Academic Press, New York. 3. Tanada, Y. 1963. Epizootiology cf infectious dis- eases. Pp. 1-27. In E. A. Steinhaus (ed.) Insect pathology: an advanced treatise, Vol. I. Academic Press, New York. 4. Watanabe, H. 1987. The host population. Pp. 71— 112. In J. R. Fuxa and Y. Tanada (eds.) Epizootiology of insect diseases. John Wiley & Sons, New York. 5. Tanada, Y. and J. R. Fuxa. 1987. The pathogen pop- ulation. Pp. 113-157. In J. R. Fuxa and Y. Tanada (eds.) Epizootiology of insect diseases. John Wiley & Sons, New York. 6. Andreadis, T. G. 1987. Transmission. Pp. 159-176. In J. R. Fuxa and Y. Tanada (eds.) Epizootiology of insect diseases. John Wiley & Sons, New York. 7. Kermack, W. D. and A. G. McKendrick. 1927. A contribution to the mathematical theory of epidemics. J. R. Statist. Soc. Ser. A. 115:700-721. 8. Waltman, P. 1974. Deterministic threshold models in the theory of epidemics. In S. Levin (ed.) Lecture notes in biomathematics. Springer-Verlag. 9. Brown, G. C. 1987. Modeling. Pp. 43-68. In J. R. Fuxa and Y. Tanada (eds.) Epizootiology of insect diseases. John Wiley & Sons, New York. 10. Anderson, R. M. and R. M. May. 1978. Regulation and stability of host-parasite population interactions. I. Regulatory processes. J. Anim. Ecol. 47:219-247. 11. Anderson, R. M. and R. M. May. 1979. Population biology of infectious diseases. Part I. Nature 280:361—367. TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3—-4) 12. Anderson, R. M. and R. M. May. 1981. The pop- ulation dynamics of microparasites and their invertebrate hosts. Phil. Trans. R. Soc. Lond. B Biol. Sci. 291:451-524. 13. Susilo, F. X. 1991. Epizootiological studies on Neozygites floridana infectious to the twospotted spider mite. Ph.D. Dissertation. University of Kentucky, Lexing- ton. 14. Susilo, F. X., R. Hasibuan, G. L. Nordin, and G. C. Brown. In press. The concept of threshold density in in- sect pathology: a theoretical and experimental study on Tetranychus—Neozygites mycosis. Proc. Congr. Insect Pathol. Yogyakarta, 12-13 Oct. 1993 (in Indonesian). 15. Anderson, R. M. 1991. Populations and infectious diseases: ecology or epidemiology? J. Anim. Ecol. 60:1- 50. 16. Platt, T. and K. L. Denman. 1975. Spectral anal- ysis in ecology. Ann. Rev. Ecol. Syst. 6:189-210. 17. Olsen, L. F. and W. M. Schaffer. 1990. Chaos ver- sus noisy periodicity: alternative hypotheses for childhood epidemics. Science 249:499—-504. 18. Brown, G. C. and R. Hasibuan. 1995. Conidial dis- charge and transmission efficiency of Neozygites floridana, an entomopathogenic fungus infecting two-spotted spider mites under laboratory conditions. J. Invertebr. Pathol. 65: 10-16. 19. STATA. 1988. Reference manual. Release 2. Com- puting Resource Center, Los Angeles, California. 20. SAS. 1982. SAS user's guide: statistics, version 5 ed. SAS Inst., Cary, North Carolina. 21. Laing, J. E. 1969. Life history and life table of Tetranychus urticae Koch. Acarologia 11:32-42. 22. Brown, G. C. In press. Theoretical and experi- mental studies of predator versus pathogen effects on spi- der mite population dynamics. In Proc. Ninth Internat. Congr. Acarol. 23. Hasibuan, R. 1991. Predator-pathogen—host pop- ulation interactions in a model laboratory system. Ph.D. Dissertation. University of Kentucky, Lexington. Trans. Ky. Acad. Sci., 56(3-4), 1995, 109-113 Foraging of Red-cockaded Woodpeckers (Picoides borealis) in Kentucky MARTINA HINES! AND PAUL J. KALISz Department of Forestry, University of Kentucky, Lexington, KY 40546-0073 ABSTRACT The 5 groups of red-cockaded woodpeckers (Picoides borealis) remaining on the Daniel Boone National Forest in 1990-1991 were observed during the winter and nesting seasons to determine the importance of different tree species as foraging substrates. On the average, red-cockaded woodpeckers foraged on hard- woods 14% of the time during the winter and 44% of the time during the nesting season. During the winter season, birds in 3 groups exhibited statistically significant preferences for foraging on pines compared to Sees (P < 0.001), and birds in 2 groups exhibited no preferences. During the nesting season, birds in 2 groups preferred hardwoods (P < 0.004), birds in 1 group preferred pine (P < 0.003), and birds in 2 groups exhibited no preferences. Southern ea oak (Quercus falcata), black oak (Q. velutina), and white oak (Q. alba) were the hardwood species used most frequently by foraging red-cockaded woodpeckers. Our results suggest that management based primarily on the removal of hardwoods to improve red-cockaded woodpecker habitat contributes to loss of biodiversity and may ultimately lead to the extirpation of the local population of the species. INTRODUCTION The northern-most population of the en- dangered red-cockaded woodpecker (Picoides borealis) occurs on the Daniel Boone National Forest in southeastern Kentucky. As of No- vember, 1994, only 4 or 5 birds in 2 groups survived. The range of this population is dis- junct from others to the south, with little pos- sibility of gene flow into Kentucky. The habitat is mixed pine-oak forests with well- developed lower canopy strata (1), vegetation typical for dry sites in the Appalachians but atypical for red-cockaded woodpecker habitat. Management of this declining red-cockaded population by the U.S. Forest Service is based on the assumption, developed in the pine for- ests of the southeastern Coastal Plain, that habitat quality for red-cockaded woodpeckers is reduced in proportion to the occurrence of hardwoods in a stand (e.g., 2). This implies that red-cockadeds prefer pines and discrimi- nate against hardwoods as a foraging substrate. To date, this assumption has not been tested in hardwood-dominated regions such as Ken- tucky. The objective of this study was to de- termine if red-cockaded woodpeckers in Ken- tucky exhibit a preference for pines as a for- aging substrate. This is an important question ! Present address: Kentucky Nature Preserves Commis- sion, 801 Schenkel Lane, Frankfort, KY 40601. because recent management by the Forest Service has been largely based on this belief, and has relied on cutting and prescribed burn- ing to remove hardwoods from active and in- active red-cockaded colony sites and other ar- eas judged potentially suitable as colony sites for the species (3). MATERIALS AND METHODS Study Area.—The study area was located in Laurel and Whitley Counties on the Daniel Boone National Forest in southeastern Ken- tucky (36°48'N, 84°18'W). This area lies along the escarpment defining the western edge of the Cumberland Plateau physiographic region and is included in the Mixed Mesophytic EOE: est region (1). The topographic heterogeneity of the land- scape explains the presence of numerous for- est types. Ridge tops are predomin: antly cov- ered with pine and mixed pine-har dwood stands. These stands are often small and occur as narrow bands along elevational contours. Common species include shortleaf pine (Pinus echinata), white oak (Quercus alba), various red oak species, and hickory (Carya spp.). ). The mid- and understories present throughout the study area are composed of red maple (Acer rubrum), sourwood (Oxydendrum aboreum), blackgum (Nyssa ie atica), and dogwood (Goins florida) (1). The lower slopes are 109 110 dominated by hardwood stands and contain a low percentage of pines. Typical species are chestnut oak (Quercus prinus), red and white oaks, yellow poplar (Liriodendron tulipifera) and beech (Fagus grandifolia). On northerly slopes and in ravines, characteristic species in- clude hemlock (Tsuga canadensis) and rho- dodendron (Rhododendron maximum). Hard- wood stands occupy approximately 65% of the area, and pine and pine-oak stands, which are considered suitable habitat for red-cockaded woodpeckers, account for the remaining 35% of the forested area (1). Field Methods.—Birds were observed at the 5 active colony sites from October to June, 1990-1991, and from December to March, 1991-1992. Four of the sites were occupied by pairs of birds, and the other site was oc- cupied by a single bird. Data were systemati- cally collected only during the 1991-1992 ob- servation period to statistically describe for- aging during the winter (December to March) and nesting seasons (April to June). During the winter period, foraging data were collect- ed during 31 field sessions, 18 during the morning (2-hour period following sunrise) and 13 during evening (2-hour period preceding sunset). A total of 311 minutes of observation was collected. During the nesting period for- aging data were collected during 19 field ses- sions, 14 during midday, 3 during the morning and 2 during the evening. A total of 426 min- utes of observation was collected. All foraging data were collected when the birds were near their cavity trees since cliffs, cut trees and brush prevented us from follow- ing the birds. During the nesting season, red- cockaded woodpeckers could be observed for relatively long periods because they spent much of their time close to the nest tree when feeding young. Birds were observed in areas that were approximately circular around cavity trees and ranged from 2.4 to 4.1 ha. When recording data, we assumed that a red-cockaded woodpecker was foraging when it probed, picked or flaked bark as described by Hooper and Lennartz (4). Birds were not considered to be foraging when active on the cavity tree around the cavity entrance or resin wells. Observations were made at 1-minute in- tervals to record the species of tree on which the bird was foraging. Observations collected in this way may be considered independent for TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) TaBLE 1. Importance (expressed as percentage of basal area and density) of tree species in red-cockaded wood- pecker colonies. Percentage Basal area Density (stems =10 (stems =2 & cm DBH) <10 cm DBH) Species Mean Median Mean Median Pinus echinata 55 57 0 0 Quercus coccinea 10 6 1 2 Carya spp. ¢ 6 7 8 Quercus alba 6 4 0 1 Quercus velutina 4 2 2 4 Pinus virginiana 4 8 0 0 Pinus rigida 0 1 0 0 Quercus stellata 3 3 1 1 Quercus falcata 2 1 5 6 Quercus prinus 2, 2 1 1 Acer rubrum 2 2 24 21 Liriodendron tulipifera 1 2 8 M7 Cornus florida 1 2 yall 26 Nyssa sylvatica 1 2) 2 2 Tsuga canadensis 1 2 6 11 Oxydendrum arboreum 1 1 12 13 Magnolia sp. 0 0 5 11 Amelanchier arborea 0 0 0 9 Carpinus caroliniana 0 0 2 4 Sassafras albidum 0 0 2 4 Fagus grandifolia 0 0 1 2 the purpose of testing the foraging prefer- ences of individual birds (M. Lacki, pers. comm.). Since the birds were not banded it was impossible to distinguish between individ- uals in groups composed of 2 birds, and, therefore, for statistical analyses, data record- ed for groups were treated as if they were re- corded for an individual bird. At each colony site, the vegetation within the area used for foraging was described. At each site, circular 0.05-ha plots were randomly located at a density of 5 per ha, and the di- ameter at a height of 1.4 m (DBH) and spe- cies of every tree 210 cm DBH was recorded. Within a 4 X 25 m strip across each circle, the number and species of stems <10 cm DBH and =2 cm DBH were recorded (Table 1). Statistical Analysis.—Chi-square tests were used to determine whether or not woodpeck- ers preferred pines or hardwoods as foraging substrate. The availability of pines and hard- woods was expressed as the percentage of the total basal area; for each site this percentage was used to calculate the expected number of foraging observations from the total number of observations. Expected values calculated in RED-COCKADED WOODPECKER IN KENTUCKY—Hines and Kalisz this way represented the case where red-cock- adeds did not discriminate between pines and hardwoods, but foraged in the two types of trees in proportion to their occurrence. Since the total population consisted of only 9 birds, and these were divided into 5 groups with members that were indistinguishable, it was impossible to collect enough independent observations to make statistical statements re- garding the foraging preferences of the entire red-cockaded population on the Daniel Boone National Forest. Statistical analyses were therefore only performed for the individual groups of birds at each site, and observations pertaining to the entire population were sum- marized, but not analyzed, statistically. The numbers of observed and expected foraging observations on both pines and hardwoods were used to calculate chi-square values for each colony site to test for preference for ei- ther tree type. Winter and nesting season data were analyzed separately. RESULTS Basal areas of stems =10 cm DBH ranged from 5 m?/ha to 13.3 m?/ha, with an average of 12 m2/ha. Pine accounted for 48 to 74% of the basal area. Shortleaf pine was the most important species in red-cockaded woodpeck- er colonies accounting for 55% of the total basal area with a range of 40 to 70% (Table 1). In contrast, pitch pine (Pinus rigida) and Virginia pine (P. virginiana), respectively, ac- counted for <1 and 4% of the basal area. Six species of oaks accounted for 27% of the total basal area and 68% of the non-pine basal area. Other individual hardwood species accounted for <2% of the basal area (Table 1). For stems <10 cm and =2 cm DBH, total stem density ranged from 90 to 650 stems/ha over all 5 col- ony sites. In contrast with conditions on the study area (Table 1), Forest Service recom- mendations for optimum red-cockaded habitat specify that hardwood density per acre should be <10/ac in the upper canopy level and <3/ ac in the mid-story (3). During both winter and nesting seasons, red-cockaded woodpeckers spent more time on shortleaf pine (74% in winter; 50% in nest- ing season) than on any other tree species. The hardwoods used by red-cockaded wood- peckers were scarlet oak (Quercus coccinea), white oak, southern red oak (Q. falcata), post 11] TABLE 2. nies of tests of red-cockaded woodpeckers’ preference for Levels of significance (P) for individual colo- pines or hardwoods (Hdwd) as foraging substrate during the winter and nesting seasons. Nesting Season Winter Season Colony P Preference P Preterence I 0.19 None <0.001 Pine II 0.003 Pine 0.096 None Ill 0.004 Hdwd >0.250 None IV <0.001 Hdwd <0.001 Pine V >0.250 None <0.001 Pine oak (Q. stellata), black oak (Q. velutina) and hickory. Red-cockaded woodpeckers were sometimes seen foraging on red maple during the winter, and on sourwood, chestnut oak and, rarely, dogwood during the nesting sea- son. Red-cockaded woodpeckers were never observed on yellow poplar, black gum, or east- ern hemlock. During the winter, 311 (86%) foraging ob- servations were on pines and 52 (14%) on hardwoods. Statistical analyses of the individ- ual colonies showed that red-cockadeds pre- ferred pine over hardwoods at 3 colonies but did not exhibit a preference at the other two colonies (Table 2). During the nesting season, 239 (56%) for- aging observations were on pines and 187 (44%) were on hardwoods. Among the 5 col- onies, foraging ranged from 34 to 89% on pines, and from 11 to 66% on hardwoods. During the nesting season the birds statisti- cally preferred pine at one colony, hardwoods at 2 colonies, and showed no preference at 2 colonies (Table 2). During the winter red-cockaded woodpeck- ers showed no apparent preference for any in- dividual hardwood species as foraging sub- strate. During the nesting season, however, red-cockaded woodpeckers, on average, uti- lized the southern red oak/black oak and the white oak/post oak (Q. stellata) groups far above their proportional occurrence in the stands. Conversely, during the nesting season, red-cockaded woodpeckers seldom foraged on scarlet oak and hickory, although these taxa were among the most important hardwoods on all sites. DISCUSSION Red-cockaded woodpeckers in our study area spent most of their time foraging on pine 112 trees, and statistically preferred pines on 3 sites during the winter season (Table 2). This result was consistent with past research (4, 5, 6, 7, 8). From previous reports of red-cock- adeds foraging on hardwoods (4, 6, 9, 10), the highest reported use was in Mississippi where birds spent 22% of their time on hardwoods (6). Our finding that during the nesting sea- son, on average, red-cockaded woodpeckers foraged on hardwoods 44% of the time, and statistically preferred hardwoods on 2 sites, contradicts the results of research in more typ- ical habitat where red-cockaded woodpeckers seem to always prefer pines over hardwoods (4). Our results support previous reports (4, 9) that red-cockaded woodpeckers exhibit a sea- sonal change in foraging preference for pines and hardwoods (Table 2). The seasonal change in preference that we documented may have been due to a shortage of arthropods on bole surfaces during the winter (11). This would force red-cockaded woodpeckers to concen- trate their winter foraging on the thick, flaky bark of pines which offers protective habitat for a large number of over-wintering arthro- pods (12). In contrast to other woodpeckers, red-cockaded woodpeckers forage chiefly by scaling and flaking bark rather than by exca- vating holes (4, 8). Flaking is especially critical during winter when most prey are hidden be- neath bark. During the nesting season, arthro- pods are probably more abundant on bole sur- faces, allowing red-cockaded woodpeckers to forage on both pine and hardwood species. The foraging behavior of red-cockaded woodpeckers in the pine-hardwood forests of the Daniel Boone National Forest is clearly different from that of red-cockadeds living in pure pine forests further south. The birds in Kentucky seem to readily use hardwoods as a foraging substrate, especially during the warmer parts of the year. This result supports prior assertions (13, 14) that red-cockaded woodpeckers are opportunistic foragers that take advantage of a wide array of food sources, and demonstrates adaptation of the local pop- ulation to the foraging habitat typical of the Appalachians. Our results have important implications for management of red-cockaded woodpeckers on the Daniel Boone National Forest, especially since the results of this one-year study were TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) in agreement with our previous observations of foraging behavior on the study area. Present management is based on regional guidelines that in turn are based primarily on research in other, ecologically very different, parts of the red-cockaded woodpecker’s range. Specific recommendations are to remove hardwoods from both the upper and lower canopy strata within areas =4 ha surrounding all active and abandoned cavity trees, and in areas designat- ed as recruitment and replacement stands; prescribed fire at 2—5 year intervals is rec- ommended to control sprouts from the stumps of the cut hardwoods (3). Implementation of these recommendations will make the species- rich and structurally-diverse forests of the area more uniform and less complex. Such man- agement contrasts with the national trend to- wards preserving and restoring natural pat- terns of biodiversity, and with the regional ten- dency towards hardwood dominance. In par- ticular, the drastic habitat alteration caused by the removal of hardwoods may cause the local population to become maladapted to its hab- itat, and may contribute to a loss of genetic diversity and ultimately to the extirpation of red-cockaded woodpeckers in Kentucky. ACKNOWLEDGMENTS This work was supported by MclIntire-Sten- nis funds and by the Daniel Boone National Forest. This is a contribution of the Kentucky Agricultural Experiment Station, Paper no. 95-09-013. LITERATURE CITED 1. Kalisz, P. J. and S. E. Boettcher. 1991. Active and abandoned red-cockaded woodpecker habitat in Ken- tucky. J. Wildl. Manage. 55:146-154. 2. Hooper, R. G., A. F. Robinson, Jr., and J. A. Jackson. 1980. The red-cockaded woodpecker: notes on life history and management. U.S. For. Serv. Gen. Rep. SA-GR 9. 3. U.S. Forest Service. 1993. Draft environmental im- pact statement for the management of the red-cockaded woodpecker and its habitat on national forests in the Southern Region. Forest Service, Southern Region, Atlan- ta, Georgia. 4. Hooper, R. G. and R. M. Lennartz. 1981. Foraging behavior of the red-cockaded woodpecker in South Car- olina. Auk 98:321-334. 5. Morse, D. H. 1972. Habitat utilization of the red- cockaded woodpecker during winter. Auk 89:429-435. 6. Ramey, P. 1980. Seasonal, sexual, and geographic variation in the foraging behavior of red-cockaded wood- RED-COCKADED WOODPECKER IN KENTUCKY—Hines and Kalisz pecker (Picoides borealis). M.S. Thesis. Mississippi State University. 7. DeLottele, R. S. and J. R. Newman. 1971. Habitat use of red-cockaded woodpeckers in central Florida. Pp. 56-67. In R. L. Thompson (ed.) The ecology and man- agement of the red-cockaded woodpecker. U.S. Dept. of Interior, Bureau of Sport Fisheries and Wildlife, and Tall Timbers Research Stn., Tallahassee, Florida. 8. Repasky, R. R. and P. D. Doerr. 1991. Home range and substrate use by two family groups of red-cockaded woodpeckers in the North Carolina Sandhills. Brimleyana esi —p2: 9. Skorupa, J. P. and R. W. McFarlane. 1976. Seasonal variation in foraging territory of red-cockaded woodpeck- ers. Wilson Bull. 88:662—665. 10. Wood, D. A. 1983. Foraging and colony habitat characteristics of the red-cockaded woodpecker in Okla- 113 homa. Pp. 51-58. In D. A. Wood (ed.) Red-cockaded woodpecker symposium IT. Fla. Game and Fresh Water Fish Comm., Tallahassee. 11. Conner, R. N. 1979. Seasonal changes in wood- pecker foraging methods: Strategies for winter survival. Pp. 95-105. In R. N. Conner, J. G. Dickson, R. R. Fleet, J. A. Jackson, and J. C. Kroll (eds.) The role of insectiv- orous birds in forest ecosystems. Academic Press, London. 12. Travis, J. 1977. Seasonal foraging in the downy woodpecker population. Condor 79:371-375. 13. Beal, F. 1911. The food of the woodpeckers of the United States. USDA Biol. Surv. Bull. No. 37. 14. Baker, W. W. 1971. Observations on the food hab- its of the red-cockaded woodpecker. Pp. 100-105. In R. L. Thompson (ed.) The ecology and management of the red-cockaded woodpecker. U.S. Dept. of Interior, Bureau of Sport Fisheries and Wildlife, and Tall Timbers Re- search Stn., Tallahassee, Florida. Trans. Ky. Acad. Sci., 56(3—4), 1995, 114-116 Extended Monitoring of Mussels (Bivalvia: Unionidae) in the Rockcastle River at Billows, Kentucky, an Historical Site RONALD E. Hour AND KAREN L. SMATHERS Division of Water, Water Quality Branch, Ecological Support Section, 14 Reilly Road, Frankfort, Kentucky 40601 ABSTRACT Surveys of unionid mussels in 1985, 1991, 1993, and 1994 in the Rockcastle River at Billows, Kentucky, yielded 18 live species. Elliptio dilatata was the most numerous, followed by Amblema plicata and Actinon- aias ligamentina. Combined, those species represented 62% of the individuals collected in all surveys. The 1991 survey showed declines (25% to 40%) in 12 of 16 species observed when compared to the 1985 survey. Those declines were sequential to increased mining activities in the Billows quadrant. During that time, Actinonaias ligamentina, Quadrula pustulosa, and Villosa taeniata remained stable with slight increases above 1985 numbers and appear to be more tolerant of intermittent sedimentation. Villosa trabalis was not col- lected live at this location. Compared with historical data, these surveys indicate that changes in species composition and shifts in abundances will continue in response to changes in habitat conditions at the Billows site. INTRODUCTION One of the objectives of the Biological Monitoring Program for the Division of Water (DOW) is to gather biological information through time. This resulted in a database that began in 1976 and continues presently, for se- lected stream sites across the state. In 1982, the Rockcastle River at Billows, Kentucky, was added to the program. This particular site had prior historical mussel data that began with the Neel and Allen survey of 1947-1949 (1). Most of the mussel collections in the drainage afterwards excluded the Billows site until the Thompson study in 1985 (2). Her study was the most thorough assessment of the Rock- castle River mussels; however, her collections were above and below the DOW site. More recently, collections from this site by Layzer and Anderson (3), yielded 8 live species. Also, Cicerello (4), observed 10 live species at Bil- lows, as part of an effort to document rare and threatened species for the United States For- est Service (USFS) in the Daniel Boone Na- tional Forest (DBNF), and for the Kentucky State Nature Preserves Commission (KSNPC) state inventory. The objective of the DOW surveys at Bil- lows was to document the resident mussel community on a continuing basis, so that changes in the community can be used (along with other components) in the evaluation of present water-quality conditions and to help characterize future trends in water quality of the Rockeastle River. Stuby SITE The DOW study site at Billows is located downstream of the SR 1956 bridge. The study site extends from the bridge to a few meters downstream of the canoe launch. This area is the upper boundary of the Rockcastle Wild River segment. A long straight reach allowed bed load materials to be deposited across the width of the river, which created a long shal- low run and a partially exposed shingle bar that extends the length of the study site (ap- proximately 75 m). The substrate is dominated by rubble and cobble-sized flat rocks mixed with larger boulders, gravels, pebbles and sands that create ample mussel habitats. METHODS All surveys were conducted between mid- June and mid-July. Instream observations and identifications were made by both authors, while one person served as the data recorder for both observers. Mussel searches were made easier by visually striking transects from bank to opposite bank across the stream. Starting from opposite banks, each observer waded toward the other on the same transect. At a meeting point about mid-stream, one ob- server stepped around the other on the down- stream side, allowing each to continue along the transect. In effect, each transect was vi- 114 MussELs IN KeENTUCKY—Houp and Smathers 115 TABLE 1. Species abundances and survey years for Rockcastle River at Billows. DOW *L&A RRC N&A 1985 1991 1993 1994 1992 1993 1964 Actinonaias ligamentina 61 66 46 ip? — 2 —_ Actinonaias pectorosa 2, 5 — 5 — 6 ( Amblema plicata 104 66 47 53 L 8 A Cyclonaias tuberculata 27 7 21 24 [L m=z A Elliptio dilatata 150 102 139 142 L 30 c Lampsilis cardium 12 20 26 5 L 10 (e Lampsilis fasciola 9 17 a 6 — 5) (; Lampsilis ovata 5 2 = — Ib == _ Lasmigona costata 1 2 + — — = Cc Ligumia recta 23 14 16 12 IL — 6 Obovaria subrotunda = il == _ = == = Pleurobema coccineum 1 4 5 8 == = (G Potamilus alatus 1 1 = 1 = 3 @ Ptychobranchus fasciolaris 74 44 39 32 = 6 © Quadrula pustulosa 12 9 ll 19 — 3 oo Strophitus undulatus 1 = = y) = == _ Tritigonia verrucosa = = — L — — Villosa taeniata 19 15 31 31 1 5 @ Villosa trabalis = = = == = _ G Totals 507 Bi7l 393 412 * Layzer & Anderson (L&A), R.R. Cicerello (RRC), Neel & Allen (N&A). Abundant (A), Common (C) sually searched for mussels twice, once by each observer moving from opposite direc- tions. That particular feature was useful be- cause it helped discover mussels that were missed in one direction by the other observer, either by being overlooked or because of sun glare and/or swnidl action on the water surface. When undisturbed mussels were located, the mussels were removed from the substrate, identified, and cleaned before replacement, so as to prevent recounting by the observer searching from the opposite direction. Starting at the downstream edge of the study site, a total of 75 transects were visually searched in an upstream direction. The transects were ap- proximately one meter wide and varied from 34 to 12 meters in width across the stream. A few mussel shells were retained in the DOW Molluse Collection. RESULTS AND DISCUSSION Eighteen species have been collected alive at this site since 1985 (Table 1). In the 1991 survey, 12 of 16 species reflected 25-40% de- clines in numbers observed (see Table 1), which correlated with the bulk (24 of 30) of permitted mining sites that began operations between 1985-1991 (5). Elliptio dilatata, the most abundant species at the site, showed a decline in 1991. Actinonaias ligamentina has , Live (L), Absent (—) steadily increased, replacing Amblema plicata as the next most numerous species. Com- bined, those species represented 62% of the total numbers of individuals collected in all surveys. Cyclonaias tuberculata, Pleurobema coccineum, Quadrula pustulosa, and Villosa taeniata populations have remained stable, with slight increases since the declines of 1991. Examples of Lampsilis ovata have been infrequent and were not observed in the last 2 DOW surveys; however, Layzer and Ander- son (3) noted its occurrence at Billows, as well as Tritigonia verrucosa, which appeared only in the 1994 DOW survey. Villosa trabalis was not observed during the DOW surv eys. Acti- nonaias pectorosa, Ouadrula pustulosa, Ligu- mia recta, and V. taeniata, although per in numbers, were observed in each survey and have shown signs of recruitment in this area. When comparing DOW data with the orig- inal Neel and Allen (1) survey, it can be con- cluded that changes in species composition and shifts in Panidanees have occurred. For instance, Actinonaias ligamentina and Quad- rula pustulosa have heconie well established at Billows, and other species, such as Lamp- silis ovata, Obovaria subrotunda, Strophitus undulatus, and T. verrucosa, have been infre- quently observed. Actinonaias pectorosa, P. coccineum, Potamilus alatus, and Lasmigona 116 costata, originally considered common to the site, were observed in very low numbers. Ex- planations for those declines include natural cycles and the effects of habitat alterations from sedimentation. The most obvious effects to the mussel community at Billows are severe shell erosion (sand blasting) in some species, and alterations in substrate compositions (ex- cessive sands). Surface mining in the head- waters (south and middle forks) were noted by Thompson (2) Layzer and Anderson (3), and Cicerello (4), but, much farther downstream and adjacent to the Billows site, (Hawk Creek drainage) there have been 30 permits issued for surface mining since 1985 (5). The permits range from 1.99 to 646 acres, totaling nearly 2,500 acres. Presently, only 5 permits are ac- tive, with disturbed lands about 1,600 acres and on the increase. If mining continues at the same rate, it could continue in the Billows quadrant (USGS) for another decade with ex- isting permits. Vannote and Minshall (7) reported that an influx of sediments appeared to be responsible for a shift in species dominance in the Salmon River Canyon, Idaho. Neel and Allen noted that E. dilatata became dominant in the Cum- berland River system sometime after Wilson and Clark’s survey (6) in 1911. Table 1 shows that E. dilatata was the most abundant species in all DOW surveys, while Actinonaias liga- mentina made slight increases in numbers ob- served each survey. Houp (8) previously doc- umented that E. dilatata replaced Alasmidon- ta marginata as the most abundant mussel in the Wild River segment of the Red River, while Actinonaias ligamentina, and Pleurobe- ma coccineum increased in abundances after the onset of coal mining in the headwaters of that eastern Kentucky stream. The above spe- cies, common to both stream systems, as well as Quadrula pustulosa and Villosa taeniata, of the Rockcastle River surveys, appear to be more tolerant to the effects of intermittent sedimentation than other mussel species in those drainages. Certainly, the timing of innate behavioral mechanisms (burrowing, and re- productive cycles) are central to individual species tolerances or intolerances of sedimen- tation. Nearly all the mussel species previously re- corded from this historical site are still pres- TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3—-4) ent. Although several species were few in number and infrequently observed, our work does validate their existence at this site and in the drainage. ACKNOWLEDGMENTS We wish to show our appreciation to fellow workers and others who have helped with field work over the years. Allen Robison, USFWS, Parrish Roush (DAQ), Gary Beck, Cliff Schneider, Giles Miller and John Brumley, DOW workers, and George Wesley Houp. Also, Dru Hawkins, for her knowledge and help with word processors, Terry Anderson, Water Quality Branch, Manager, DOW and Dr. G. A. Schuster, EKU, for helpful manu- script reviews and Mike Mills, Ecological Sup- port Section, Supervisor, DOW, and Dr. B. A. Branson, EKU, for their positive support. We thank them all. LITERATURE CITED 1. Neel, J. K. and W. R. Allen. 1964. The mussel fauna of the upper Cumberland basin before its impoundment. Malocologia 1:427-459. 2. Thompson, Y. L. 1985. The mussel fauna of the Rockcastle River system, Kentucky (Bivalvia: Unionidae). M.S. Thesis, Eastern Kentucky University, Richmond, Kentucky, U.S.A. 3. Layzer, J. B. and R. M. Anderson. 1992. Impacts of the coal industry on rare and endangered aquatic organ- isms of the upper Cumberland River Basin. Final report submitted to Kentucky Department of Fish and Wildlife Resources, Frankfort, Kentucky, U.S.A., and Tennessee Wildlife Resources Agency, Nashville, Tennessee, U.S.A. 4. Cicerello, R. R. 1993. A survey of the Unionids (Bi- valvia: Unionidae) of the Rockcastle River, Middle Fork to Billows, Kentucky. Kentucky State Nature Preserves Commission, Technical Report, Frankfort, Kentucky, U.S.A. 5. Department of Surface Mining Reclamation and En- forcement. 1995. Surface Mining Information System, Div. of Permits, #2 Hudson Hollow, Frankfort, Kentucky 40601. 6. Wilson, C. B. and H. W. Clark. 1914. The mussels of the Cumberland River and its tributaries. United States Fish Commission, United States Bureau of Fisheries Doc- ument No. 781:1-63. 7. Vannote, R. L. and G. W. Minshall. 1982. Fluvial processes and local lithology controlling abundance, struc- ture, and composition of mussel beds. Proc. Nat. Acad. Sci. 79:4103-4107. 8. Houp, R. E. 1993. Observations on long-term ef- fects of sedimentation on freshwater mussels (Mollusca: Unionidae) in the North Fork of Red River, Kentucky. Trans. Ky. Acad. Sci. 54(3-4):93-97. Trans. Ky. Acad. Sci., 56(3-4), 1995, 117-127 Mossbauer Study of Single Crystal Biotite and Phlogopite AMER S.LAHAMER AND Curis ].LANGLEY Physics Department, Berea College, Berea, KY 40404 ABSTRACT At room temperature, Mossbauer spectra of single crystal biotite and phlogopite were found asymmetric quadrupole doublets. This asymmetry is largest when the angle between the gamma-ray and the normal to the surface of the absorber is 0°, and it vanishes at about 55°. Based on previous work in the literature, the biotite spectra have been fitted with 2 lorentzian doublets. presumably due to Fe?* of lorentzian doublets, 2 on M1 and M2 sites, while the phlogopite spectra have been resolved into 3 sets due to Fe?* on M1 and M2 sites and the third possibly due to Fe®* on M2 sites. This work is intended to answer the question of how many doublets can be used in fitting a certain spectrum. In some of the previous work, assignments of doublets seem somewhat arbitrary due to their overlap. One possible way of determining the number of doublets of Fe? and Fe** to be used in fitting a spectrum is by changing the oxidation state of iron in some samples, using heat treatment without changing their crystal structures. INTRODUCTION Some of the questions about minerals con- taining iron are: in a given mineral, how many iron sites are there, in what oxidation state, and what is the relative population of these states? Such questions are important, for ex- ample, in learning about the conditions of rock-formation. Methods such as wet chemistry are not completely reliable in answering the above questions. For one thing, in wet-chemistry ex- periments, the sample is usually destroyed as a result of the method, unlike Méssbauer spectroscopy where the sample is unaltered after the study is done. If one ignores the pos- sibility of different sites having different re- coil-free fractions, then Méssbauer spectros- copy lends itself reasonably well to identifying the states of Fe2* and Fe°* ions and the crystal structure of their surroundings. In micas, these states are represented through doublets in a Mossbauer spectrum. The number of doublets that could be fitted in a complicated Méssbauer spectrum is still uncertain. A complicated spectrum here means that there exists an overlap of doublets. Some of these doublets are for Fe2* and some are for Fe**, both of which might be in the same or different sites. Mossbauer spectra of biotite and phlogopite single crystal micas were taken at different ori- entations and different thicknesses, all at room temperature. We fitted the biotite data with 2 sets of doublets. The phlogopite spectra were fitted with 2 and with 3 sets of doublets. Our biotite results are consistent with what is found in the literature. But in the phlogopite spectra, the Fe** doublet is small and buried under the Fe2+ doublet. It is not clear at all that the Fe*+ exists. The sign of the electric field gradient (efg) for Fe?* is known to be opposite of that of the Fe®* ion. So, we annealed the mica at low enough temperature that the Fe?* loses one electron and becomes an Fe** ion without changing the crystal structure of the sample. Then, by taking different spectra at different orientations it is possible to determine the number of sites and the oxidation states of iron occupying those sites, as shown below. X- ray analysis was done on the sample before and after the heat treatment as a way of in- suring the crystal structure did not change during the heat treatment. METHOD The intensity ratio of the two quadrupole components is given by we =e Si) se cost) (1) (2 + 3 sin?@) where @ is the angle between the efg principal axis (z-axis) and he gamma-ray propagation direction (Fig. 1), and where the inte nsity ra- Ibi Ze 118 surface normal direction gamma direction A >? : . . . s Se ee * . s —~ s . | c Fic. 1. Geometry of the experiment. tio is obtained using the areas under the ab- sorption peaks. For polycrystals (powders), the 2 lines have equal intensities (ie., R = 1) because of the averages of: i 2 (sin?0) = — | sin?@ sin 8 d@ dd = -—, At 3) and 1 ; 1 (cos?0) = — | cos?0 sin 0 d@ db = —. AT 3 For single crystals, R = 5/3 when the gam- ma-ray is perpendicular (8 = 90°) to the prin- cipal axis of the electric field gradient, R = 1/3 when the gamma-ray is parallel (8 = 0°) to the efg axis, and R = | for the magic angle 0 = 54.7°. However, if the recoilless radiation is anisotropic and the recoil-free fraction f is itself a function of 0, then the ratio becomes | (1 + cos?0) £(@) sin 0 dé preset Wet cd | (2/3 + sin?0) £(0) sin 8 dé = F[f(6)] 4 1. (2) In other words, an anisotropic recoilless frac- tion should produce an asymmetry in the +(3/2) — +(1/2) quadrupole doublet. The above effect was discovered by Goldanskii (1). By measuring R (ratio of areas under the peaks) as a function of the angle, 0, in single crystals, it is possible to determine which of TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) the spectral lines corresponds to the +(3/2) > +(1/2) transition and which to the +(1/2) > +(1/2) transition. In this way, the sign of the quadrupole interaction constant (e?Qq) is de- termined. The sign of eQ of the Méssbauer nuclide is usually known from other nuclear techniques (ie., eQ = +0.18 X 10-* cm? for the 14.4 keV of *Fe nuclide). Then the sign of eq (=V,,) can be extracted. For example, the meaning of a positive sign is that the + (3/2) energy level is higher in energy than that of the #(1/2) level of the *’Fe nuclide as shown in Figure 2. The above method is under the assumption that the experimenter knows the number of doublets and the oxidation states of the iron in advance before meaningful analysis can take place. Previous Mica STUDIES Many types of mica have been studied ex- tensively by many authors. A review of Méss- bauer data on trioctahedral micas was done by Darby (2). The review was done in order to determine the existence of tetrahedral Fe?+ and cation ordering in micas and to find rea- sonable ranges of the hyperfine parameters of different Fe sites. Hargraves and Rancourt (3) studied single crystal phlogopite mica at room temperature and at different orientation angles. They claimed that thickness effects are not negligi- ble, the Fe2+ and Fe** area ratios can not be obtained due to the overlap, and the resolu- tion of the cis and trans components of the Fe?’ is at best questionable. Darby (4) did a Méssbauer and wet-chem- istry study on 52 biotite samples. In one set of data, which she called O-L-10, she fitted the data with 4 doublets, 2 for Fe2* and 2 for Fe>*, all in the octahedral sheet, and she fitted the same data with 2 doublets for the Fe?+ and 1 doublet for octahedral Fe®* and the other for the tetrahedral Fe®*. She claimed that “even by visual inspection” the latter was a better fit. After examining her spectra care- fully, it was not clear to us how such conclu- sions can be made. Rancourt (5) studied 6 trioctahedral micas to determine the spectral features of '!Fe**. Similar studies were also done by Darby (6). But that aside, after examining their spectra, it is still hard to assign such specific oxidation MossBavuER Crystal Stupy IN Kentucky—Lahamer and Langley (+3/2) Fe f=3/2 Eo (+1/2) r= 1/2 Isomer Shift = “J D ee = i) | SS he sew o ja - Velocity Fic. 2. Quadrupole splitting in “Fe with I = 3/2 in the excited state and I = 1/2 in the ground state. Top: The I = 119 m, (Vzz > 0) £3/2 +1/2 +1/2 Quadrupole Splitting 0 + Velocity 3/2 level is split into two sublevels by positive electric quadrupole interaction. The ground state is not split because there is no quadrupole moment in a nucleus with I = 1/2. The levels of I = 3/2 and I = 1/2 are shifted by electric monopole interaction (giving rise to the isomer shift, 8). Bottom: Resultant Mossbauer spectrum (schematic). states for the iron. Méssbauer spectroscopy alone is not quite enough to determine the existence of '/Fe*+ or 'lFe®+ in mica since they overlap under '!Fe?*+. By looking into their spectra carefully at liquid nitrogen tem- perature (77 K), it seems that resolution was not that much better than that of the room- temperature spectra. It would have been more informative to use liquid helium. In 1993, Darby (7) wrote an article com- menting on Rancourt et al.’s work and sug- gesting that powder samples were more suited than single crystals for determining the oxi- dation states of iron. In the same journal issue, 100000 90000 ; 95 mg/cem’2 80000 100000 900007 61 mg/cm‘’2 80000 100000 45 a2 ooo] een Transmission 80000 100000 19 mg/cm42 90000 80000 70000 6 -4 » ETes 3: Rancourt responded by reiterating that single crystals are better suited and offer more in- sight (8). In this study, we tried to address all of the above issues and to examine some of the claimed results. We have performed Méss- bauer spectroscopy of single crystals as well as powder biotite and phlogopite for different orientation and different thicknesses. We only heat-treated the phlogopite since it is the con- troversial mica. We have done powder X-ray diffraction studies on the phlogopite before and after the heat treatment. The results of this study are outlined in the next section. TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) 0 y 4 6 mm/s Mossbauer spectra of biotite of different thicknesses all at a = 0°. Data ACQUISITION AND ANALYSIS Biotite A standard Méssbauer spectrometer was utilized in collecting the data. The source used was Co in a rhodium matrix. The absorber used in the present study was single crystal biotite and phlogopite mica [K(Mg,Fe), (OH,F),AISi,O,]. The biotite comes in dark sheets (flakes). About | centimeter by 1 centimeter square of biotite was cut for an absorber which was weighed and then mounted on a polarimeter, and data were accumulated for 20 hours at MossBavER Crystal Stupy IN Kentucky—Lahamer and Langley ial ————— 100000 fv awventne = he é eroetigsorn 3 * 98000 a= 96000 980004 x9 =| =| : 96000 ; 100000 980007] «= 40° 100000 es 1G): 98000 96000 -6 -4 -2 0 2: 4 6 mm/sec Fic. 4. Mdéssbauer spectra of biotite at different orientation angle a. each orientation angle. Six sheets were peeled off and stacked consecutively for different thickness measurements. Since we did not know the exact direction of the efg in advance nor, did we know the appropriate thickness, we measured the trans- mission rate as a function of the angle be- tween the direction of the gamma-ray and the perpendicular to the surface of the absorber which we call a (Fig. 1). A typical spectrum of the results of thickness experiments for bi- otite in this case is shown in Figure 3. After the appropriate thickness was select- ed, different spectra at different orientation angles were taken (Fig. 4). The biotite data were fitted using a least squares fitting routine with 4 single lorentzian lines. By graphing the widths of the lines as a function of the orien- tation angle, it seems that line 1 and line 4 are coupled while line 2 is coupled to line 3 (Fig. 5). Such an approach was first introduced by Hargraves and Rancourt (3). The results of the fitting routine using the above doublets are summarized in Table 1 From Table 1 we see that R.,, quite equal to the theoretical ratio R,,(0 which may mean that the principal axis of o efg is not perpendicular to the surface of the (a) is ie Widths (Arbitrary Units) a (degrees) Fic. 5. for line 3, @ for line 2 Widths versus angle. © for line 4, © for line 1, @ sample. It is clear though, from the system- atics of Figure 4 and Table 1, that the left peak corresponds to the +(3/2) — +(1/2) transi- tions, which means that the sign of the inter- action e?qQ is negative. The sign of eQ for °"Fe is known to be positive, implying that the sign of the efg (eq) is negative. Here, negative efg means that the +(1/2) energy level is high- er than the +(3/2) energy level in the quad- rupole splitting. Phlogopite The phlogopite we studied came in 2 colors: one is light in color (iron poor) and the other is dark (iron rich). The present results are for the dark phlogopite only. Again, a 1 centime- ter by 1 centimeter square of phlogopite was cut for an absorber which was mounted on the polarimeter after being weighed, and data were accumulated for 20 hours at each ori- entation angle. For the thickness experiments, TABLE 1. TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3—4) a stack of these squares was used. The inten- sity of the quadrupole lines has been mea- sured as a function of the angle, a, between the normal to the surface and the direction of the gamma-ray (Fig. 1). Typical spectra for thickness and angular orientation measure- ments for the phlogopite are shown in Figures 6 and 7. The summary of the fitting routine for the phlogopite is shown in Table 2. Here, the data were fitted with 3 sets of doublets. Two dou- blets were attributed to Fe2+ on M1 and M2 sites and the third to Fe*+ on the M2 site. Again, from Table 2 we see that R,,,(@) is not quite equal to the theoretical ratio R,,(8) for Fe?*. It is clear, though, from the system- atics of Figure 7 and Table 2 that the left peak corresponds to the +(3/2) > +(1/2) transi- tions, which means that the sign of the inter- action e?qQ is negative. The sign of eQ for °*7Fe is known to be positive from other nucle- ar techniques, implying that the sign of the efg (eq) is negative for Fe?+. Notice that the Fe*+ doublet is buried under the Fe2+ doublet. The suggested doublet for the Fe** had the right peak larger than the left peak which implied that the sign of the efg of Fe** is positive. As can be seen from the last column of the table, the fit was good. Other hidden doublets may be assumed but are difficult to justify. The phlogopite can also be fitted with only 2 doublets yielding the same interpretations as that of biotite. The obtained Méssbauer fit pa- rameters for Fe?* on M2 site are: A = 2.54 mm/sec and 6 = 1.07 mm/sec, while A = 2.08 mm/sec, and 6 = 1.08 mm/sec for Fe?* on the M1 site. An illustrative spectrum is shown in Figure 8. The misfit is the same as that of the biotite and is of the order of 0.2% which is within the statistical error. Results of the fitting routine to biotite. A is the quadruple splitting and 6 is the isomer shift with respect to iron metal. RHWHM stands for the ratio of the half widths at half maximum. M1 and M2 refer to two sites. RHWHM Rex Rexp a Rth (M1 (M2 M1 M2 0° 3.00 2.60 2.24 1245) 0.88 10° 2.83 2.39 23 1.06 0.64 20° 2.40 2.18 2.08 1.03 0.75 30° 1.91 2.00 1.92 1.00 0.84 40° 1.47 1.40 1.55 1.138 0.88 50° 1.13 IE 1.25 1.17 0.91 60° 0.88 0.60 0.81 1.14 0.76 5 (mm/sec) A (mm/sec) ee ee ee Misfit M1 M2 M1 M2 (%) 1.04 1.03 1.98 2.47 0.098 1.04 1.06 2.15 2.69 0.451 1.03 1.06 2.15 2.73 0.423 1.06 1.04 2.06 2.50 0.186 1.05 1.03 2.10 2.50 0.334 1.05 1.04 2.08 2.50 0.295 1.00 1.02 2.06 2.49 0.262 MOossBAUER CrysTAL StuDY IN KENrUcKy—Lahamer and Langley 100000 + 130 mg/cm“%2 81 cm/mg’2 49 cm/mg*2 16 mg/cm’2 Fic. 6. We annealed chosen sheets of the phlogo- pite at different temperatures in air and slow- cooled or quenched in room temperature, wa- ter, or liquid nitrogen. We found that the method of quenching had no effect on the spectra. The temperature range that produced clean spectra is between 600°C and 800°C. Typical spectra of annealed phlogopite are shown in Figure 9. The Méssbauer spectra of Figure 9 are fit- ted with two doublets. It is clear from the quadrupole splitting values in Table 3 that all of the Fe2+ is converted to Fe?+. Notice the quadrupole splitting of the Fe+ in the un- treated phlogopite is much lower than for the annealed one. Based on this it is safe to as- 123 + 0 Zz 4 6 mm/sec Mossbauer spectra of phlogopite at different thicknesses all at a = 0°. sume that the Fe** in the untreated phlogo- pite is too small to be statistically significant. From Figure 9, it is clear that the sign of efg of the Fe** ion is opposite of that of Fe?* ion. We did powder X-ray diffraction studies on the phlogopite before and after the heat treat- ment where the results were shown in Figure 10. By looking to the X-ray spectrum, it seems the heat treatment did not change the crystal structure significantly on the lone range scale. CONCLUSIONS The sign of the electric field eq (V,,) was found to "he negative for Fe?* in biotite and phlogopite and to be positive for Fe?* in 124 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) [ra a Pa RTE Te CS 100000 petersenii peenttcwetinrteweats \ / VU reume. 7 poreriortraecne 97500 100000 +-tystfentnelispa Mpa, tans tee ee 975004 a= 20° 100000 A ee v2 i hatte fy eeglt ~~ 3 + + + $ + ¢? + mission 97500) %= Trans +4 teehee +e * t¢ + + ue “ed et Pat, +t ee 100000 Fite ittrentnesateaie a ght sevens atta a= 60° 97500 + % ¢ 95000 -6 -4 -2 0 2 4 6 mm/sec Fic. 7. Méssbauer spectra of phlogopite at different orientation angle a. TABLE 2. Summary of the results of the fitting routine to phlogopite. A is the quadrupole splitting and 6 is the isomer shift with respect to iron. M1 and M2 refer to two sites. Fe2+ 5 (mm/sec) A (mm/sec) snake Bee Go eos lectin 3+/2+ el Rexp Rexpyi. i Se Rexp 8 A Ratio Misfit a Rth (M1 (M2) M1 M2 M1 M2 (M2) M2 M2 M2 (%) 0° 3.00 2.71 2.69 1.01 1.04 2.24 2.59 0.20 0.26 0.30 0.07 0.339 20° 2.40 2.09 2.39 1.04 1.03 2.13 2.54 0.21 0.26 0.30 0.09 0.456 40° 1.47 E33 1.42 1.07 1.04 1.93 2.49 0.62 0.26 0.30 0.04 0.388 60° 0.88 1.02 1.03 1.02 1.02 2.63 2.10 0.70 0.26 0.30 0.06 0.159 MOSSBAUER CRYSTAL STUDY IN KENTUCKY 101000 100000 tH 99000 5S 98000 = “v BA € 2 97000 © he = 96000 95000 94000 Fic. 8. Phlogopite spectra at a = 0° fitted with only two doublets. *. e+ + 4H + + -2 phlogopite. Negative quadrupole interaction results in the +(1/2) energy level being higher than the +(3/2) level which is consistent with the present understanding in the literature. We were able to get a good fit of the data of the biotite with only two doublets which we attribute to Fe2* on the M1 and M2 sites re- ++ mm/sec Lahamer and Langley spectively. We saw no evidence of Fe** in our biotite sample. This is also consistent with most results in the literature. We measured the gamma-ray absorption as a function of thickness for both biotite and phlogopite in order to determine the appro- priate size (Fig. 3 and 6). We found the ap- TaBL_E 3. Summary of the results of the fitting routine to the annealed phlogopite. A is the quadrupole splitting and 6 is the isomer shift with respect to iron metal. This is interpreted as Fe** at both sites. Rexp a Rth (M1) 0° 0.33 0.65 20° 0.42 0.69 40° 0.68 0.89 60° 1.14 ell? Rexp (M2) 0.67 0.70 0.90 1.16 RHWHM M1 1.00 1.00 1.00 1.00 M2 1.00 1.00 1.00 1.00 5 (mm/sec) M1 M2 0.285 0.292 0.271 0.302 0.280 0.292 0.295 0.307 A (mm/sec) M1 M2 0.859 0.559 0.877 O57 0.863 0.551 0.882 0.552 Misfit (%) 0.386 0.677 0.551 0.439 126 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) 100000 Wie Ca 90000 100000 a = 20° 90000 =| : = 100000 a = 40° ; 90000 100000 a = 60° 90000 80000 4G 4 2 0 2 4 6 mm/sec Fic. 9. Méssbauer spectra of annealed phlogopite at different orientation angle a. propriate range is of the order of 10-to-30 mg/ cm?. The isomer shift and the quadrupole split- ting values of Fe?* for both samples are con- sistent with those found in the literature (Ta- bles 1 and 2). The corresponding values for Fe?* are not consistent. We believe that the amount of Fe?* in our original (unannealed) phlogopite was not enough to be detected. If there had been enough Fe** present, the quadrupole splitting values for Fe®+ would have been enhanced while the positions of its doublets would have stayed in the same channel numbers. Instead, the quadrupole splitting values and their po- sitions of the doublets were different from those of the Fe** in the unannealed phlogo- pite. The question about the existence of Fe** with different coordination, namely [4] and[6], probably can be resolved if the experiment could be done at liquid helium temperatures which might separate the Fe** and Fe** dou- blets further through the magnetic phase tran- sitions. We believe this technique can be applied to a variety of iron-containing samples to find the true oxidation states of the iron. ACKNOWLEDGMENTS This research is supported partially by a grant from the Keck Foundation and from the John Auxier Fund to the Physics Department of Berea College. We thank Mr. Larry Lipchinsky, Geology Department at Berea College for supplying us MOossBavER CrystaL Stuby IN Kentucky—Lahamer and Langley 127 10 Intensity (Arbitrary Units) 10 20 30 Fic. 10. with the mica. We thank Dr. Glenn Julian from the Physics Department of Miami Uni- versity in Oxford, Ohio; Dr. S. Jha from the Physics Department at the University of Cin- cinnati, Cincinnati Ohio; Dr. R. A. Dunlap, the Department of Physics at Dalhousie Uni- versity, Halifax, Nova Scotia, Canada; and Dr. Smith T. Powell III, Berea College, Berea, Kentucky for their useful discussions. LITERATURE CITED 1. Greenwood, N. N. and T. C. Gibb. 1971. bauer spectroscopy. Chapman and Hall, London. 2. Darby, Dyar M. 1987. A review of Méssbauer data M6ss- Phlogopite before annealing Phlogopite after annealing 70 60 40 50 20 (deg) Powder X-ray diffraction pattern observed for the phlogopite samples. on trioctahedral micas: evidence for tetrahedral Fe** and cation ordering. American Mineralogist 72:102—112. 3. Hargraves, P. and D. G. Rancourt. 1990. Single- crystal Méssbauer study of phlogopite mica. Can. J. Phys. 68:128-144. 4. Darby, Dyar M. 1990. Méssbauer spectra of biotite from metapelites. American Mineralogist 75:656-666. 5. Rancourt, D. G. 1992. Méssbauer spectroscopy of tetrahedral Fe** in trioctahedral micas. American Min- eralogist 77:34-43. 6. Darby, Dyar M. et al. 1986. study of ferruginous one-layer trioctahedral micas. Amer- ican Mineralogist 71:955—965. 7. Darby, Dyar M. and D. G. Rancourt. 1993. Mdéss- otros in trioctahedral Mossbauer spectral bauer spectroscopy of tetrahedral Fe** discussion. American Mineralogist 78:665—668. micas Trans. Ky. Acad. Sci., 56(3-4), 1995, 128-133 Nematodes Found in the Opossum (Didelphis virginiana) and Four Other Species of Mammals in Central Kentucky in 1991 EUGENE T. LYONS, SHARON C. TOLLIVER, AND SHELBY STAMPER Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546-0099 AND BRAD SNYDER Southeastern Cooperative Wildlife Disease Study, College of Veterinary Medicine, University of Georgia, Athens, GA 30602 ABSTRACT Opossums (n = 50), striped skunks (n = 3), eastern cottontail rabbits (n = 2), muskrats (n = 2), and a coyote, collected on horse farms (n = 14) in central Kentucky in the fall of 1991, were examined for internal helminths. All parasites (nematodes, cestodes, and trematodes) recovered were counted. Only nematodes were identified. Species of nematodes recovered from opossums were Capillaria spp., Physaloptera turgida, Strongyloides spp., Longistriata didelphis, Viannaia viannai, and Cruzia americana; striped skunks were Gongylonema spp., Physaloptera maxillaris, Baylisascaris columnaris, and Strongyloides spp.; eastern cot- gy PP Yysaloy ] gy PP tontail rabbits were Obeliscoides cuniculi, Trichostrongylus calcaratus, and Trichuris leporis; muskrats were Physaloptera spp. and Trichostrongylus spp.; the coyote (only small intestine examined) were Molineus bar- batus, and Uncinaria stenocephala. INTRODUCTION Several species of wild mammals were col- lected in central Kentucky in 1991 on farms where leptospirosis had been found in horses. The purpose was to determine if they were carriers for this disease in horses. Collecting the mammals provided an opportunity to ex- amine some of them for internal parasites. Mostly opossums, and lesser numbers of skunks, eastern cottontail rabbits, muskrats, and a coyote, were examined for helminths. Total counts were made for all parasites re- covered, but only nematodes were identified. MATERIALS AND METHODS A total of 50 opossums (Didelphis virgini- ana), 3 striped skunks (Mephitis mephitis), 2 eastern cottontail rabbits (Sylvilagus floridan- us), 2 muskrats (Ondatra zibethica), and 1 coyote (Canis latrans) were collected from 14 horse farms in central Kentucky between No- vember 8 and December 12, 1991 for parasi- tologic examination. They were trapped by personnel from the Southeastern Cooperative Wildlife Disease Study, University of Georgia. The age/sex categories were: opossums—38 adults and 12 juveniles/26 males and 24 fe- males; skunks—2 adults and 1 juvenile/1 male, 1 female, and | sex not recorded; eastern cot- tontail rabbits—2 adults/] male and 1 female; muskrats—I1 adult and 1 juvenile/2 females; coyote—1 adult male. Organs examined for parasites were: lungs (including the trachea), liver, esophagus, stom- ach, small intestine, and large intestine. Snips of the lungs, liver, and esophagus were re- moved for other researchers before a parasi- tologic examination was done. Therefore, some parasite specimens from these tissues, which were examined in fresh condition for parasites with a stereoscopic microscope (at about 30), may have been “lost.” The gas- trointestinal tract was separated into stomach, small intestine, and large intestine, including cecum. Contents were emptied into separate containers and then each segment was washed several times with water. Rinses were added to the appropriate container of contents and this material was preserved with 5% formalin. The gastrointestinal tract was then opened with scissors and any parasites observed by gross examination were removed. Next, each portion of the gastrointestinal tract was placed in containers of artificial digestive juice (1% pepsin and 1% HC1 in an incubator at 37°C) for about 3 hours; then, the mucosal side was 128 NEMATODES IN KENTUCKY MamMmMats—Lyons et al. 129 TABLE il. Data on internal parasites recovered from opossums in a survey in central Kentucky. No. of parasites Opossums (n = 50 Parasites Range Mean* Total No. infected (%) infected Nematodes Capillaria spp.** Lungs M — 3.0 3 ] (2) F Q-7*** al 46 22 (44) M&F 0-7 DH) 49 22 (44) G.i. tract M 0-S 2a 30 14 (28) F 0-11 Sul 68 2D (44) M&F 0-11 3.9 98 25 (50) Physaloptera turgida**** Imm 24,300 474.8 23,265 49 (100 M 3-53 18.4 §99 49 (100 F 2-79 20.0 978 49 (100) M&F 6-131 38.3 ESian 49 (100 Total 10-4,314 518.4 25,918 50 (100 Strongyloides spp. 0-42 15.0 45 3 (6 Longistriata didelphis M 0-23 4.7 85 18 (36 Longistriata/Viannaia F 0-21 (eS 198 27 (54 Viannaia viannai M 0-2 1.7 5 3 (6 Cruzia americana Imm 0Q—1,800 63.1 POA 36 (72 M 0-242 62.7 2,821 45 (90 F 0-305 75.9 3,568 47 (94 M&F 0-547 135.9 6,389 48 (96 Total 0-—1,804 180.4 8,660 48 (96) Imm-—Unidentified 0-39 6.8 129 19 (38 Total Nematodes 12-6,119 703.7 35,187 50 (100 Cestodes 0-510 114.4 1,487 13 (26 Trematodes 0-421 69.1 2.419 35 (70 * Mean = value for infected opossums; ** Capillaria spp.—tfrom lungs, probably C. didelphis and from gastrointestinal tract, probably C. longicauda; *** Sex of one specimen not determined, but included data with females; **** Includes 776 from one opossum for which only total number counted; M = male; F = female; Imm = Immature. washed under running water while being rubbed by hand to remove loosened material. This material and the digest were preserved with 5% formalin; the remainder of the walls _ was discarded. Digestion was done to free par- asites associated with the walls and mucus. For the single coyote collected, only the small intestine was examined for parasites. The fixed contents, water rinses, and digests from the gastrointestinal tract were washed into a series of sieves (10, 20, 40, 60, and 100 mesh). Residue was examined for helminths with the aid of a stereoscopic microscope at about 30. Total counts of all helminths were done. Nematodes were identified by usage of various reference publications (1-29). Ces- todes and trematodes were not identified. RESULTS AND DISCUSSION For the opossums (Table 1), the 2 most prevalent species of nematodes were Physa- loptera turgida (100%) and Cruzia americana (96%). The high prevalence of both of these species is similar to that reported by several other investigators (4, 11, 18, 30-32). Num- bers of male versus female specimens per in- fected animal ranged (mean number/infected opossum) from 3 to 53 (18.4) and 2 to 79 (20), respectively, for P. turgida and from 0 to 242 (62.7) and 0 to 305 (75.9), respectively, for C. 130 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) TABLE 2. Data on number of opossums infected with internal parasites relative to farm where collected. Internal parasites Nematodes No. ae Rae Physal- Stdes. Im. Longi- Farm no. exam Lungs GI tract optera spp. Un-ID striata** —Viannaia Cruzia Cestodes Trematodes No. of opossums infected 1 3 1 2 3 0 1 3 0 3 0 2 2 3 2 2 3 0) 0 2 1 2 1 DA 3 2 0 0 2 0 0 1 0 oy 0 0 4 5 2 2 5 0 3 0 0 5. ] 2 5 1 0 0 1 0 1 0 0 1 0 1 6 6 2, 5 6 1 4 6 0 6 D 6 ¢ 1 1 0 1 0 1 0 0 1 0 1 8 3 1 ) 3 0 3 1 0 2 1 2 9 4 3 1 4 0 3 1 0 4 0 4 10 4 2 1 4 0 3 1 0 4 0 a) 11 1 0 0 1 0 0) 1 0 1 0 1 12 3 2 0 3 0 0 0 0 3 0 3 13 8 2 ¢ 8 1 0 6 2 8 1 6 14 6 4 5 6 1 0 6 0) 6 4 5 Total 50 22 25 50 3 19 28 3 48 13 35 (T) inf. (44) (50) (100) (6) (38) (56) (6) (96) (26) (70) * Complete names—Physaloptera turgida; Strongyloides spp; Im. = Immature (Unidentified); Longistriata didelphis, Viannaia viannai, Cruzia americana. **.Longistriata didelphis category includes L. didelphis males and longistriata/Viannaia females. americana. Immature specimens (determined only by much smaller size than adults) varied in number (mean) from 2 to 4,300 (474.8) for P. turgida and 0 to 1,800 (63.1) for C. ameri- cana. The 2 next highest prevalent nematode species were Longistriata/Viannaia females (54%) and Capillaria spp. (probably C. longi- cauda) (50%) from the gastrointestinal tract. Longistriata didelphis males were found in 36%. of the opossums. Two other reports in- dicated higher prevalence (18, 30) for L. di- delphis. For C. longicauda, lower infection rates were found in 2 studies (4, 18) than for Capillaria spp. from the gastrointestinal tract in the present study. The number of male L. didelphis varied (mean) from 0 to 23 (4.7) and of female Longistriata/Viannaia from 0 to 21 (7.3). Capillaria spp. males vs. females were 0 to 8 (mean = 2.1) and 0 to 11 (3.1), respec- tively. Most of the Longistriata/Viannaia fe- male specimens are probably L. didelphis be- cause so few male V. viannai were found. The females were grouped together because of similarities in features. Other nematode spe- cies recovered (prevalence) were: from the lungs—Capillaria spp. (probably C. didelphis) (44%), and from the intestines—unidentified immatures (38%), Strongyloides spp. (6%), and Viannaia viannai (6%). Prevalency for Capillaria didelphis was less in 1 study (32) and more in another (18) compared to that of Capillaria spp. in the lungs of opossums cur- rently investigated. Strongyloides spp. (21) and Viannaia viannai (14) are apparently un- common in opossums. The highest total num- ber of nematodes in an individual opossum in the present study was 6,119; it had the great- est number of immature P. turgida and C. americana. Cestodes (0 to 510; mean = 114.4) were recovered from 26% of the opossums; trema- todes were highly prevalent (70%). The num- ber of specimens of trematodes varied from 0 to 421 (mean = 69.1) per opossum. Data on the number of opossums infected with inter- nal parasites on each farm are recorded (Table 2). Prevalences of the various species or types of parasites were highest on Farm No. 6. The 3 skunks examined (Table 3) were all infected with the nematodes Physaloptera maxillaris, Baylisascaris columnaris, and Strongyloides spp.; Gongylonema spp. were found in 1 skunk. Cestodes were present in all 3 individuals, but trematodes in none. Preva- lence of P. maxillaris in several other studies varied greatly (16, 33-35). Baylisascaris co- lumnaris were much more prevalent in 2 oth- er studies (34, 35). Strongyloides spp. were NEMATODES IN KENTUCKY MAMMALS—Lyons et al. TABLE 3. Internal parasites recovered from skunks, eastern cottontail rabbits, muskrats, and a coyote. No. of parasites Animals infected Parasites Range Mean* Total No (%) Skunks (n = 3) Nematodes Gongylonema spp. M — 1 ] 1 (33) F — 1 1 1 (33) M&F ]—] 2 2 1 33) Physaloptera maxillaris Imm 295-1,828 921.67 2.765 3 (100) M 30-53 39.00 LEZ, 3 (100) F MO", 80.00 240 3 (100) M&F 102-150 119.00 357 3 (100) Total 400-1,978 1,040.67 OuE22 3 (100) Balisascaris columnaris M 1-18 7.00 21 3 (100) F 3-9 6.67 20 3 (100) M&F 4-27 13.67 4] 3 (100) Strongyloides spp. 1-93 38.33 115 3 (100) Cestodes 10-92 39.67 119 3 (100) Eastern cottontail rabbits (n = 2 Obeliscoides cuniculi M 3-4 3.50 4 2 (100) F 1=t 1.00 2 2 (100) M&F 4-9 4.50 9 D, (100) Trichostrongylus calcaratus Bao) 2.00 4 2 (100) F 4-6 5.00 10 2 (100) M&F 6-8 7.00 14 2 (100) Trichuris leporis M — 1.00 1 1 (50) Cestodes = 3.00 3 1 (50) Trematodes — 2.570 2.570 1 (50) Muskrats (n = 2) Nematodes Physaloptera spp. (Imm) }—} 1.00 2 2 (100) Trichostrongylus spp. M — 3.00 S IL (50) F — 4.00 4 1 (50) M&F 34 3.50 ih 1 (50) Cestodes _— 2.00 2 il (50) Trematodes 37-3,758 189.75 3,795 2 (100) Coyote (n = 1) Molineus barbatus M — 3.00 3 1 (100) F — 5.00 5 1 (100) M&F 35) 8.00 8 ] (100) Uncinaria stenocephala M — 6.00 6 i (100) F — 2.00 2, 1 (100) M&F 2-6 §.00 8 1 (100) Cestodes = 39.00 39 1 (100) * Mean = value for infected animals. Imm = Immature; M = male; F = female. 131 132 found to be less prevalent by 2 researchers (34, 36). Gongylonema longispiculum were of low prevalence in | study (36). For both cottontail rabbits examined (Table 3), 2 nematode species, Trichostrongylus cal- caratus and Obeliscoides cuniculi, were recov- ered. One rabbit was also infected with Tri- churis leporis. Cestodes and trematodes were present in 1 rabbit. Four other reports (2, 17, 31, 37) indicated greatly variable prevalence of O. cuniculi and T. calcaratus in this species of rabbit. For T. leporis, 3 investigators reported (2, 17, 37) lower infection rates than in the current survey. The 2 muskrats examined (Table 3) were infected with the nematodes, Physaloptera spp. (immature) and Trichostrongylus spp. In addition, cestodes were recovered from | muskrat and trematodes from both muskrats. Physaloptera spp. have been previously found (23). Trichostrongylus calcaratus were found in a few animals in 2 studies (5, 20). In the small intestine of the coyote, the nematodes, Molineus barbatus and Uncinaria stenocephala, were present. Also, cestodes (not Echinococcus spp.), but no trematodes, were found. Both of the species of nematodes were evident previously in low numbers of coyotes (10, 24). This survey of internal parasites gave an in- dication of species, prevalence, and numbers in some of the wild mammals in the central Kentucky area. Especially meaningful are data from opossums for which a much greater number were examined than for the other hosts. Also, further information was obtained for Kentucky, supplementing previous surveys for helminths in muskrats in Madison County (20) and in eastern cottontail rabbits in west- ern Kentucky (37). ACKNOWLEDGMENTS The investigation reported in this paper (No. 95-4-072) was made in connection with a project of the Kentucky Agricultural Exper- iment Station and is published with the ap- proval of the director. LITERATURE CITED 1. Anderson, R. C. 1992. Nematode parasites of ver- tebrates, their development and transmission. CABI, Uni- versity Press, Cambridge, England (Distributed U.S.A— The University of Arizona Press, Tucson), 578 pp. TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) 2. Andrews, C. L. and W. R. Davidson. 1980. Endo- parasites of selected populations of cottontail rabbits (Syl- vilagus floridanus) in the southeastern United States. J. Wildl. Dis. 16:395—401. 3. Babero, B. B. 1957. Some helminths from Illinois opossums. J. Parasit. 43:232. 4. Babero, B. B. 1960. Further studies on helminths of the opossum, Didelphis virginiana, with a description of a new species from this host. J. Parasit. 46:455—-463. 5. Beckett, J. V. and V. Gallicchio. 1967. A survey of helminths of the muskrat, Ondatra z. zibethica Miller, 1912, in Portage County, Ohio. J. Parasit. 53:1169-1172. 6. Butterworth, E. W. and M. Beverley-Burton. 1977. Capillaria didelphis n. sp. (Nematoda:Trichuroidea) from the opossum, Didelphis virginiana L. in Georgia. Can. J. Zool. 55:616-619. 7. Chandler, A. C. 1932. A new species of Longistriata (Nematoda) from the cotton rat, Sigmodon hispidus, with notes on the division of the Heligmosominae into genera. J. Parasit. 19:25-31. 8. Chandler, A. C. 1933. Notes on the helminth par- asites of the opossum (Didelphis virginiana) in southeast Texas, with descriptions of four new species. Proc. U.S. Nat. Mus. 81:1-15. 9. Chandler, A. C. 1942. The helminths of raccoons in East Texas. J. Parasit. 28:255—268. 10. Conti, J. A. 1984. Helminths of foxes and coyotes in Flordia. Proc. Helm. Soc. Wash. 51:365-367. 11. Crites, J. L. 1956. A redescription of Cruzia amer- icana, a nematode parasitic in the opossum, Didelphis marsupialis virginiana. J. Parasit. 42:68—72. 12. Dikmans, G. 1932. A new nematode worm, Vian- naia bursobscura from the opossum, with a note on other parasites of the opossum. Proc. U.S. Nat. Mus. 79:1-6. 13. Dikmans, G. 1935. New nematodes of the genus Longistriata in rodents. J. Wash. Acad. Sci. 25:72-81. 14. Dikmans, G. 1943. The occurrence of Viannaia viannai Travassos (Nematoda:Heligmosomidae) in opos- sums in North America. Proc. Helm. Soc. Wash. 10:6—7. 15. Dyer, W. G. 1969. Helminths of the striped skunk, Mephitis mephitis, in North America. Am. Midl. Nat. 82: 601-605. 16. Erickson, A. B. 1946. Incidence of worm parasites in Minnesota Mustelidae and host lists and keys to North American species. Am. Midl. Nat. 36:494-509. 17. Erickson, A. B. 1947. Helminth parasites of rab- bits of the genus, Syvilagus. J. Wildl. Mgt. 11:255-263. 18. Feldman, D. B., J. A. Moore, M. W. Harris, and J. L. Self. 1972. Characteristics of common helminths of the Virginia opossum, (Didelphis virginiana) from North Carolina. Lab. An. Sci. 22:183-189. 19. Graybill, H. W. 1924. A new species of roundworm of the genus Trichostrongylus from the rabbit. Proc. U.S. Nat. Mus. 66:1-3. 20. Harley, J. P. 1972. A survey of the helminths of the muskrat Ondatra z. zibethica Miller, 1972, in Madison County, Kentucky. Tran. Ky. Acad. Sci. 33:13-15. NEMATODES IN KENTUCKY MAMMALS—Lyons et al. 21. Little, M. D. 1966. Seven new species of Stron- gyloides (Nematoda) from Louisiana. J. Parasit. 52:85—97. 22. Morgan, B. 1941. A summary of the Physalopter- inae (Nematoda) of North America. Proc. Helm. Soc. Wash. 8:28—30. 23. Penn, G. H., Jr. 1942. Parasitological survey of Louisiana muskrats. J. Parasit. 28:348—349. 24. Seesee, F. M., M. C. Stemer, and D. E. Worley. 1983. Helminths of the coyote (Canis latrans Say) in Montana. J. Wildl. Dis. 19:54—55. 25. Skrjabin, K. I, N. P. Shikhobalova, and R. S. Shul'ts. 1954. Trichostrongylids of animals and man. In K. I. Skrjabin (ed.) Essentials of nematodology, Vol. III. Izdatel’stvo Acad. Nauk. S.S.S.R., Moscow. Israel Program for Scientific Translations, Jerusalem, 1960. 26. Skrjabin, K. I., N. P. Shikhobalova, R. S. Shul'ts. 1954. Dictyocaulidae, Heligmosomatidae and Ollulanidae of animals. In K. I. Skrjabin (ed.) Essentials of nemato- dology, Vol. IV. Izdatel’stvo Acad. Nauk. $.S.S.R., Moscow. Israel Program for Scientific Translations, Jerusalem, 1971. 27. Skrjabin, K. I., A. A. Sobolev, and V. M. Ivaskin. 1967. Spirurata of animals and man and the diseases caused by them. Thelazioidea. Part 4. In K. I. Skrjabin (ed.) Essentials of nematodology, Vol. XVI. Izdatel’stvo Acad. Nauk. S.S.S.R., Moscow. Israel Program for Sci. Translations, Jerusalem, 1971. 28. Travassos, L. 1937. Revisao da familia Trichostron- gylidae Leiper, 1912. Monographias do Instituto Oswaldo Cruz, Rio de Janerio. 133 29. Yorke, W. and P. A. Maplestone. 1926. The nem- atode parasites of vertebrates. P. Blakiston’s Son & Co., Philadelphia, 536 pp. 30. Gray, J. B. and R. C. Anderson. 1982. Observa- tions on Turgida turgida (Rudolphi, 1819) (Nematoda: Physalopteroidea) in the American opossum (Didelphis virginiana). J. Wildl. Dis. 18:279-285. 31. Holloway, H. L., Jr. 1966. Helminths of rabbits and opossums at Mountain Lake, Virginia. Bull. Wildl. Dis. Assoc. 2:38—39. 32. Miller, G. C. and R. Harkema. 1970. Helminths of the opossum (Didelphis virginiana) in North Carolina. Proc. Helm. Soc. Wash. 37:36-39. 33. Dyer, W. B. 1970. Helminths of the striped skunk, Mephitis mephitis Schreber, in North Dakota. Proc. Helm. Soc. Wash. 37:92-93. 34. Goldberg, A. 1954. Parasites of skunks in the Beltsville, Maryland area. Proc. Helm. Wash. Soc. 21:29- 34. 35. Mead, R. A. 1963. Some aspects of parasitism in skunks of the Sacramento Valley of California. Am. Mid. Nat. 70:164—167. 36. Babero, B. B. 1960. A survey of parasitism in skunks (Mephitis mephitis) in Louisiana, with observations on pathological damages due to helminthiasis. J. Parasit. 46:26-27 (Abstr.). 37. Strohlein, D. A. and B. M. Christensen. 1983. Metazoan parasites of the eastern cottontail rabbit in western Kentucky. J. Wildl. Dis. 19:20-23. Trans. Ky. Acad. Sci., 56(3—4), 1995, 134-137 Index Herbariorum Kentuckiensis III RONALD L. JONEs, Davip A. EAKIN, AND Ross C. CLARK Department of Biological Sciences, Eastern Kentucky University, Richmond, KY 40475 ABSTRACT A survey conducted during the academic year 1994-1995 provides information on herbarium collections at 13 Kentucky institutions. Totals of approximately 243,000 vascular plant specimens and 2,700 nonvascular plant specimens are reported for the state. INTRODUCTION The objective of this study was to determine the current status of collections of vascular and nonvascular plants in the state. This is an update of previous surveys published by Las- setter (1) and Jones (2). In the earlier report, 119,000 specimens were reported from 12 herbaria, and in the latter study 187,000 spec- imens were reported from 15 institutional her- baria. For this update, a survey was conducted by mail and by phone. All known Kentucky herbaria at public and private institutions were included. Private collections were excluded. The following data were requested: name and address of collection, phone/fax/e-mail num- bers, emphasis and special collections, loan and exchange policies, and names of curators. In addition, information was gathered on the status of data-basing efforts. We appreciate the cooperation of all who supplied informa- tion. RESULTS This survey presents information on 13 in- stitutional herbaria. Totals of approximately 243,000 mounted vascular plant collections and 2,700 nonvascular plant collections are re- ported. Vascular plant collections at the Uni- versity of Kentucky (two herbaria) and the University of Louisville number 101,000; at the 5 regional institutions there are about 122,000 specimens; collections at the 5 private colleges total approximately 20,000. The great majority of private college collections are at Berea College. More than half of nonvascular plant collections are at the University of Ken- tucky. Data gathered in this survey are listed be- low. Acronyms (given in parenthesis after the institution) and general format are based on Index Herbariorum (3). Proposed but unpub- lished acronyms are indicated by an asterisk. BEREA: Herbarium of Berea College (BE- REA), Biology Department, Charles Martin Hall Science Building, Rm 212B, Berea, KY 40404. 606-986-9341, ext. 6320. Fax: 606-986- 4506, e-mail: Ralph-F.Thompson@Berea.Edu. Established about 1961. 15,000 vascular plant specimens. Special interest in Fabaceae and Campanulaceae of Kentucky, surface-mine flo- ra, and general floristics of Kentucky. Collec- tions of J. D. Pittillo, D. D. Taylor, and J. R. Abbott. On-going projects: floras of Madison and Laurel Counties. No loans, visitation open to qualified researchers. Ralph L. Thompson, Curator. BOWLING GREEN: Herbarium of Western Kentucky University (WKU), Biology Depart- ment, Rm 218 Thompson Complex NW, Bowling Green, KY 42101. 502-745-6008. Fax: 502-745-6856. e-mail: murreze@wkuvxl.wku. edu. Established in 1967. 15,000 vascular plant specimens, teaching collection of 600 fungi. Special interests in Calycanthus, Cor- nus, molecular and morphological data in un- derstanding the evolutionary history of plants, and local and pathogenic fungi. Collections of E. O. Beal, R. Athey, and G. Johnson. On- going projects: Systematics of Cornus, vari- ability in the internal transcribed spacer region (ITS) of nuclear ribosomal DNA, demograph- ics of Spiraea virginiana. Standard loan and exchange policies. Kenneth A. Nicely, Zack E. Murrell, Jeff Jenkins, Curators. CAMPBELLSVILLE: Biological Collection, Science Building Rm 101, Campbellsville Col- lege, Campbellsville, KY 42718. 502-465-8158. Established in 1965. About 1,000 vascular plant specimens. Flora of Taylor County and 134 HERBARIA IN KENTUCKY—Jones et al. surrounding counties. Exchanges and loans available. G. Weddle, M. Rogers, Curators. GEORGETOWN: Herbarium of Georgetown College, Department of Biological Sciences, Georgetown, KY 40324. 502-863-8085. e-mail: brafaill@gtc.georgetown.ky.us.edu. Estab- lished in 1945. 1,000 vascular plant specimens. General floristics. Standard loan and exchange policies. Barbara Rafaill, Curator. HIGHLAND HEIGHTS, Herbarium of Northern Kentucky University (KNK), De- partment of Biological Sciences, Highland Heights, KY 41099. 606-572-6390. Fax: 606- 572-5639. Established in 1973. 26,500 vascular plant specimens. Special interests and current research in Carex, Poaceae, cultivated plants, general flora. Standard loan and exchange pol- icies. Robert F. C. Naczi, John W. Thieret, Cu- rators. LEXINGTON: Herbarium, College of Agri- culture, Department of Agronomy, University of Kentucky, Agricultural Science Building North Rm A-4, Lexington, KY 40546. 606- 957-4898. Fax: 606-257-2185. e-mail: jgreen@ca.uky.edu. Established about 1887. About 21,000 vascular plant specimens. Em- phasis on agricultural weeds of Kentucky. Many specimens of Harrison Garman and Mary Didlake from 1896-1930. A public ser- vice collection, providing identifications of specimens sent to the College. No exchange or loans: visitors welcome. {i Ds,Creen, Cuz rator. LEXINGTON: Herbarium of the University of Kentucky (KY), Thomas Poe Cooper Build- ing, Department of F orestry, Lexington, KY 40546. 606-257-7596. Fax: 606-323-1031. e- mail: forl21@ukcc.uky.edu. Reestablished in 1948 after fire destroyed the previous collec- tion. Estimated 50,000 vascular plant speci- ~ mens, 1,500 bryophytes. Flora of the Blue- grass region and general floristics of Kentucky. Collections of Mary Wharton and_ various county floras. Standard loan and exchange pol- icies. Robert Paratley, Curator. LOUISVILLE: Davies Herbarium, Depart- ment of Biology, University of Louisville (DHL), Louisville, KY 40292. 502-852-5940. Fax: 502-852-0725. e-mail: wsdavi01@ulkyvm. louisville.edu. Established in 1953. 30,000 vas- 135 cular plant specimens. Floristics of nearby counties and natural areas. Research collec- tions of Malacothrix (Asteraceae). Standard loan and exchange policies. W. S. Davis, Cu- rator. MOREHEAD, Herbarium of Morehead State University (MOKY*), Lappin Hall, Room 306, Morehead, KY 40351. 606-783-2947. Fax: 606- 783-5002. e-mail: h.setser@msuacad.more head-st.edu. Established in 1930s. 12,000 vas- cular specimens, 455 bryophytes. Data-base project on-going. Flora of Eastern Kentucky, Rowan County and adjacent counties, Neo- tropical Tiliaceae. Fern collections of T. Mc- Coy. Current research on local wetlands and flora of Rowan County. Standard loan and ex- change policies. Howard L. Setser, Curator. MURRAY, Herbarium of Murray State Uni- versity (MUR), Department of Biological Sci- ences, Blackburn Science Building, 5th floor, Murray, KY 42071. 502-762-2786. Established in 1967. 35,000 vascular plant specimens; 600 nonvascular plants. Data-base project on-go- ing. Special interest and current research in flora of the Jackson Purchase. Fern collections of T. McCoy, and recently acquired the Athey Herbarium, formerly located in Paducah, and many Athey specimens from Memphis State University. Standard loan and exchange poli- cies. Marian J. Fuller, Curator. RICHMOND: Herbarium of Eastern Ken- tucky University (EKY), Department of Bio- logical Sciences, Memorial Science Rm 170, Richmond, KY 40475. 606-622-6257. Fax: 606-622-1020. e-mail: bioclark@acs.eku.edu. Established in 1974. 33,000 vascular plant specimens; 500 bryophytes. Special interests and current research in woody plants, wetland flora, Asteraceae, Aquifoliaceae, regional county floras, rare species studies, statewide generic treatments, and bryophytes of Ken- tucky. Recent acquisitions include a set of Mary Wharton collections and specimens from the Kentucky State Nature Preserves Com- mission. Sets of specimens from several nat- ural areas and local county floras. Interested in exchanges of vascular plants and bryophytes of southeastern United States. Standard loan policies. Ronald L. Jones, Ross C. Clark, and David A. Eakin, Curators. 136 WILLIAMSBURG: Herbarium of Cumber- land College, Biology Department, Science Building 116, Williamsburg, KY 40769. 606- 539-4399. Fax: 606-539-4490. e-mail: tyet- ter@cc.cumber.edu. Established in 1984. 2,000 vascular plant specimens. Special inter- est in Lamiaceae. Standard loans and ex- change policies. Todd Yetter, Curator. WILMORE: Herbarium, Department of Bi- ology, Asbury College, Wilmore, KY 40390. 606-858-3511, ext. 2233. Fax: 606-858-3921. Established in 1967. 350 vascular plant spec- imens. Herbaceous flora of Jessamine County. No exchange or loans. John Brushaber, Cura- tor. DISCUSSION The current estimated total of 243,000 vas- cular plant specimens held in 13 Kentucky in- stitutions represents an increase of 56,000 specimens in the past 8 years. About 70% of this increase occurred at the regional public universities. It should be noted that some of this increase was due to the incorporation of old collections, i.e., Wharton and Athey spec- imens, and therefore not all was the result of new collecting. The number of institutions with herbaria has decreased by two since the previous survey. In addition, with collections now at West Virginia University (S. Studlar, pers. comm.), there is no longer a bryophyte herbarium at Centre College in Danville, and the number of nonvascular plant collections in the state has decreased by 50%. Other changes since the 1987 survey (2) are the presence of new curators at Western Ken- tucky University, Northern Kentucky Univer- sity, Georgetown College, University of Ken- tucky (both herbaria), Richmond (2 additional curators), and Williamsburg. The KY Herbar- ium is being transferred from the School of Biological Sciences to the Department of For- estry. The EKY Herbarium recently has been moved into a larger remodeled facility; storage capacity has approximately doubled. Fax num- bers and e-mail addresses are now generally available. The only active data-basing projects are at Morehead and Murray Universities, but several other curators plan to begin such pro- jects in the future. The Kentucky State Nature Preserves Commission (502-573-2886) main- tains a database for their rare plant records, as TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) well as a small reference collection. Discus- sions have already started among curators on the possibilities of sharing databases generated at different Kentucky herbaria over the Inter- net. The total number of collections in Kentucky herbaria is still far below that of nearby states. For example the 1990 edition of Index Her- bariorum (3) lists over 800,000 vascular plant specimens in 8 herbaria in Tennessee, over 259,000 in 8 herbaria in West Virginia, over 700,000 in 11 herbaria in Indiana, over 400,000 in 22 herbaria in Virginia, and over | million vascular plant specimens in 14 Ohio herbaria. The discrepancy is even greater for nonvascular collections. For example there are 22,500 mosses and liverworts at the University of Cincinnati (CINC) (J. Snider, pers. comm.), and 165,000 nonvascular cryptograms at the University of Tennessee (TENN) (D. Smith, pers. comm.), compared to the 2,700 in Ken- tucky collections. There has been considerable bryophyte collecting in the state over the last two decades, but the majority of these collec- tions have been deposited outside the state, particularly in CINC and TENN. There are also many Kentucky vascular plant specimens housed in other states—a list of herbaria with considerable Kentucky holdings was given by Jones (2). Obviously much collecting remains to be done in Kentucky, in particular, many counties in east-central and south-central Kentucky which have received little attention from collectors. There is a need to investigate these and many other areas in Kentucky in order to gain a better understanding of the presence and distribution of vascular and non- vascular plant species. An atlas of the state vas- cular flora is now a possibility in the foresee- able future and, perhaps soon thereafter, a manual. Relatively little is known about Ken- tucky’s nonvascular plants, but with new stud- ies of these plants now occurring at several institutions, available information is likely to grow rapidly. In this survey, some curators expressed con- cerns about the future of their institutional collections. Others thought there should al- ways be a place for even small research and teaching herbaria, especially when the value of these collections for varied data-gathering ac- tivities (from molecular to floristic to ecologi- cal to geographic) is recognized. Another com- HERBARIA IN KENTUCKY—lJones et al. ment was that, if departments and institutions appreciate the critical role of organismal bi- ology in broad biological training, then system- atics and biological collections will continue to be valued. In summary, this survey indicates to us that (1) active improvement of the documentation of Kentucky's flora is occurring, primarily by regional public institutions and private insti- tutions which retain active systematists on their faculties; (2) Kentucky’s flora remains much more poorly documented than that of most southeastern states; (3) Kentucky's col- lections of nonvascular plants are particularly depauperate; (4) there is an increasingly ur- gent need for additional, active documentation ef Kentucky plants; and (5) factors and a co- operative spirit are emerging that could lead to significant improvement in coverage and in- 137 formation sharing i in the near future. Of major significance is the fact that in the last 3 years 3 new systematic botanists have been added to the faculties of the public regional univer- sities. All curators participating in this survey expressed a willingness to cooperate, contrib- ute, and support our common goal of increas- ing our knowledge of the flora and vegetation of Kentucky. LITERATURE CITED 1. Lassetter, S. 1978. Index Herbariorum Kentuckien- sis. Trans. Ky. Acad. Sci. 39:147-149. 2. Jones, R. 1987. Index Herbariorum Kentuckiensis II. Trans. Ky. Acad. Sci. 48:84—87,. 3. Holmgren, P. K., N. H. Holmgren, and L. C. Bar- nett. 1990. Index Herbariorum. Part I. The herbaria of the world, 8th ed. New York Botanical Garden, Bronx, New York. Trans. Ky. Acad. Sci., 56(3—4), 1995, 138-140 FORUM Biodiversity and Kentucky’s Heritage Ross C. CLARK Department of Biological Sciences, Eastern Kentucky University, Richmond, Kentucky 40475 It is crucial to the future of our mental and physical health, and that of our species, that we decide what we think about biodiversity and what we will try to do about preserving or destroying it. Human progress has never been shaped by those who are comfortable with quick, simple answers, and it should not be otherwise today in Kentucky. Those who are willing to address the issue thoughtfully are those who should call the tune for the fu- ture. Before European settlement, Kentucky was a place where resources were shared by all Indian tribes close enough to benefit—not the domain of any single tribe. The current status of Kentucky as a commonwealth implies the same thing: Kentucky’s natural resources should benefit all its citizens. Even though our heritage of English common law permits us to hold title to land and its resources and to do largely what we please with them, it is ecolog- ically impossible to own anything in this world. From a functional viewpoint, we simply use what is in this world to survive and reproduce, and. then pass out of the picture. We do not own the world. It owns us. Money is an artificial and very incomplete currency. Leaving aside societal values, the only real currency in the world—the only things of fundamental value—are the energy and nutrients that support the metabolism of organisms, including ourselves. Modern civi- lization uses money to value the extraction and circulation of resources, but does not use money to value the end products of resources. If we were consistent about how we use mon- ey, we would use it to value all aspects of re- source extraction and use, including the integ- rity of the atmosphere, water resources and habitat alteration and loss. But we do not live in a world where the use of natural resources is valued completely. Our species is raping the world, and every one of us is participating. All of us use more than we need. None of us recycles our waste products so that all of them can be reused by someone or something else. What does this have to do with biodiversity? Biodiversity is an issue because the activities of our species threaten it. What does it mat- ter? Why should we care? There are two fun- damental reasons why we should care. One reason is practical and one is ethical. The practical reason why we should care about preserving biodiversity is that we are the product of diverse ecosystems. A couple of years ago, I asked students in one of my class- es to write down where they would live and what their surroundings would be like, if they could live anywhere in the world they desired. Every person wrote that he or she wanted to live simply in the country, with trees, wild an- imals, clean air and clean water. Those students didn’t choose their pre- ferred surroundings by accident. For more than 99% of humans’ existence on earth, we lived in general equilibrium with the ecosys- tems we inhabited. We have practiced agri- culture and lived in large settlements for only 15,000 years out of the 2+ million years our genus has existed. The makeup of our brains was not shaped in cities. It was shaped in na- ture. Biologically, our real home is nature, not towns and cities. It is not coincidental that we need diversity of landscape, diversity of habi- tat, diversity of plants and animals around us to be mentally healthy individuals and cul- tures. The main values to us of tropical rain- forest or mixed mesophytic forest are not the economic products or medicines they may contain. The most fundamental value of intact ecosystems and their organisms is that without them, we do not know who we are. Without a connection to nature we cannot actually feel, we are strangers in a strange land. We should not have to lose our connection with nature to discover the obvious truth. The second main reason we should care 138 FORUM about biodiversity is ethical. Ethics is the property of being positively influenced by the needs of others, of keeping the welfare of oth- ers in mind. Whether one is ethical or not is not as fundamental as where our natural home is. We cannot choose, and no one can choose for us, whether or not we are children of na- ture; we just are, because it happened that way. One can choose whether or not to be ethical; or, whether we feel responsibility to- ward others may be determined by how we are raised, or by what happens to us during our lives. Ethical people and societies will feel some responsibility toward the needs of other species that share the earth. If we are consci- entious, we will try to live lightly on the earth, so that other species also ii experience a de- cent quality of life. They did not choose to be here with us. They are at our mercy. If we are ethical, we will save them a share of a decent living. If we are not, we will not care whether they exist or not, or will always place what we want above their well-being. If we are ethical, we will actively preserve diversity in ecosys- tems which all organisms require to get their food and raise dhein young. What does all this mean to people who say “no one has the right to tell me what I can do with my land?” What does it mean to the cor- poration claiming a right to pollute with im- punity because it gives people jobs, or the cor- poration claiming the right to destroy thousands of acres of diverse forest to make chipboard or run a biomass electrical gener- ating station? For Kentucky, the answer should be clear. Has Kentucky forgotten how the timber bar- ons from the monthicast gave its citizens poorly paid temporary jobs and shipped its magnifi- cent forests (and most of the money) some- where else? Has Kentucky forgotten how the coal barons did the same thing with its coal, leaving the State with a legacy of black lung, dead streams, ruined property and poor peo- ple? How many times must we see the process repeated to know that people “doing whatever they want to with their land” is an idea that doesn't benefit the Commonwealth in the long run? This Commonwealth is responsible for the welfare of its people. Public officials are elect- ed and appointed to serve that welfare, not to serve their own interests. We don’t live as is- 139 lands; we live together. When the common good must be served, private property “rights” must give way. Those “rights” are not inalien- able. They were invented by humans. For the common good, traditional rights are often lim- ited by humans for the benefit and future of all, including other species. If individuals are ecologically unethical, then state regulations can help to impose ethics. Bad habits are hard to break, but for the good of all, Kentucky must change. Independence, an admirable quality, is the only thing some Kentuckians have left to be proud of. What would help persuade people that it is in their own best interest to give up some of that independence? Perhaps more people would be willing to give up some in- dependence if they could feel worthwhile some other way, if they could see the hope of a better future, of assured, sustainable jobs that did not require the destruction of the landscape which they value. Kentuckians say they are proud of their state. They love the mountains, lakes and rivers. W hat if they could sell that pride to outsiders and make a decent living doing it? What if Kentucky once again got serious about attracting tourists by repairing and upgrading its state park facili- ties? What if people could work in secondary timber industries, exporting finished products instead of sawlogs and chipboard? What if they could retrane houses for energy conser- vation and solar assisted heating instead of burning whole forests to generate more elec- tricity? What if state gov ernment could work very hard to attract nonpolluting industries, in- stead of plowing the same old eroded resource extraction-ecosystem destruction furrow? But what about unskilled people who can- not handle sophisticated jobs? Consider South Carolina. About 30 years ago, South Carolina built a system of technical chook to give its citizens access to the real jobs of the inane. When a major corporation considers locating to South Carolina, the tech schools sit down with company management and work carefully with the industry to design a special curricu- lum to train workers to work in that industry. The state and the people pay the training costs. By the time the industry has built its facility, there are plenty of well-trained work- ers available for the specific jobs the industry offers. That’s how Spartanburg came to have 140 the highest concentration of foreign compa- nies doing business of any location in the Southeast. It’s also how South Carolina landed several Michelin facilities and Mercedes Benz. It’s also how my stepson, without a college ed- ucation, learned how to be a technical drafts- man and worked his way up to being project manager in a major construction firm, making more money than I do. He loves his work. It’s rewarding. He’s proud of himself. He should be. South Carolina’s vision made his success possible. The same thing could happen in Kentucky. There were only two prerequisites for the South Carolina success story: people who are willing to work, and a government dedicated to their future. It took vision and serious in- vestment in education on all levels. With the constructive insistence of its en- lightened citizens, more Kentuckians could someday be proud of something more than just being Kentuckians. Proud of a_ better, more livable environment; proud of better TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) jobs; proud of healthy ecosystems that remind visitors where all of us came from. Proud of a progressive government which anticipates the future. All these things are intertwined. You can't separate them from each other. To pro- duce a success story, all these factors must be addressed at the same time for a sustained pe- riod. The reason we should work to create bet- ter jobs and lives in the midst of resolutely preserved biodiversity is because it is the right thing to do for ourselves and our children. Does Kentucky have the vision and the guts to do what is right for the future of the people of this Commonwealth, or must we remain an increasingly degraded resource extraction col- ony for the rest of the nation? Will timber tide us over so coal can tide us over so Maxie Flats can tide us over so oil can tide us over so bio- mass and chipboard plants can tide us over so chicken processors can tide us over, or is there a better way? Think, Kentucky. Think Kentucky. Long- term independence and pride come from hav- ing it all together and keeping it that way. Trans. Ky. Acad. Sci., 56(3-4), 1995, 141 NEWS AND COMMENTS ANNUAL MEETINGS The annual meeting of the Kentucky Acad- emy of Science for 1995 will be a joint affair with the Tennessee Academy of Science, 16, 17, 18 November, in Bowling Green, Ken- tucky. All members should make an effort to attend this forum. The 1996 meeting will be at Kentucky State University in Frankfort, and the 1997 meeting will be at Morehead State University. Epitror’s Apios It has been my pleasure to serve the Acad- emy as Editor of the Transactions for the last 15 years or so, mostly to good purpose, I trust. But all good things, as the old saw goes, must come to an end. I would be remiss if I failed to acknowledge all the great help I have had from many people, including John Thieret, Vince DiNoto, Tom Green, Bob Naezi, and many others. And, of course, Varley E. Wei- deman who, through all those years, served faithfully as Index Editor. I wish our new Ed- itor, John Thieret of Northern Kentucky Uni- versity, good luck and peace in his duties. And finally, since I have you as a captive audience, I wish to make a few parting com- ments. During the last 30 years technological advancement has been nothing if not astound- ing. Yet, our attention to natural history has failed to keep pace. Those among us who have scant interest in organisms above the cellular and sub-cellular level tend to view ecologists, naturalists, macro-evolutionists, and popula- tion geneticists as relicts from some long-past era. Not true. It has become patently clear, at least to me and other field biologists, that if there is going to be any salvation of the natural world it will have to come from the camp of the naturalist, not the technological one. We cannot let our love affairs with machines cause an erosion of scientific insight into ecological matters. Machines like computers are fantastic in their abilities to urge along the thought pro- cesses that are involved in deducing from ev- idence, but they are little else. Computers can never replace contemplative consideration of the nature of things. We need well-educated naturalists today more than we have ever needed them in the history of science, but we are not educating enough of them. Not train- ing—educating. And all this from a life-long naturalist who is not apt to change his thought patterns— Branley Allan Branson. 141 Trans. Ky. Acad. Sci., 56(3-4), 1995, 142-146 INDEX TO VOLUME 56 2-Chloroethyl ethyl sulfide, 15, 18— 19 3-seed mercury, 4 a-fetoprotein repressor region, 91 molecular dissection of, 91 ABSTRACTS OF ANNUAL MEETING PAPERS, 80-95 ACADEMY AFFAIRS, 55-61 Acalypha rhomboidea, 4 Acer rubrum, 109, 110 Achillea millefolium, 4 Acipenser guldenstadti, 25 Acipenser transmontanus, 22 Acremonium coenophialum, 2 Actinonaias ligamentina, 114, 115, 116 Actinonaias pectorosa, 115 Aeshnidae, 43 Ag-chemicals, effect on human erythrocytes, 87 Agrimonia pubescens, 4 Agrimony, 4 Alasmidonta marginata, 116 Allium sp., 4 Alzheimer’s Disease, relationship of protease nexin-1, 91-92 Amblema plicata, 114, 115 Amblyomma americanum, 47 Ambrosia artemisiifolia, 4, 6 Ambystoma mexicanum, 94 Amelanchier arborea, 110 ANDERSON, B., 93 Andropogon virginicus, 4 Anisoptera, 41 ANNUAL MEETING PROGRAM, 62-79 Antennaria plantaginifolia, 4 Anti-cancer drugs, 91 novel synthesis of, 91 Antigens, in immune privileged sites, 88 lymphocyte recognition, 88 ANTONIOUS, G., 80, 81, 83 Apocynum cannabinum, 4 Apoptosis, in F9 teratocarcinoma cells, 83 Arachidonic acid metabolism, 92 Are Kentucky students sleeping enough?, 94 Argia fumipennis violacea, 44 A. moesta, 44 A. tibialis, 44 A. translata, 41, 44 A. sedula, 44 Asclepias syriaca, 4 A. tuberosa, 4 Ash, white, 4 Asimina triloba, 81 Aster, 4 Aster ericoides, 4 A. patens, 4 AVDIUSHKO, SERGET, 91 Axolotl lateral line, neuromast re- generation of, 93 Axolotl salamander, 94 clycoconjugate expression in, 94 lateral line sensory epithelia of, 94 BACKER, CASSANDRA K., 91 BAKER, ANGIE, 90 BAKER, N., 80 BALAK, K. J., 93, 94 BALDSIN, B., 83 Balisascaris columnaris, 131 Barbarea sp., 4 BARKSDALE, JAMES B., 87 BARTH, M. M., 89 BASHAM, BRENT, 82 Basiaeschna janata, 41, 43, 44 BASS, MICHAEL J., 92 Baylisascaris columnaris, 128, 130 BEDEL, ALVIN, 82 Beech, 110 BIEK, DONALD P., 90 Big milkweed, 4 Billows, Kentucky, an historical site, 114-116 Biodiversity and Kentucky's Heri- tage, 138-140 Biotite, Méssbauer study of, 117— WEA Bivalvia: Unionidae, 114-116 Black cherry, 5 Black locust, 5 Black oak, 109, 111 Black-eyed susan, 5 Blackberry, 5 Blackgum, 109 Blackhaw, 5 BLAIR, MATT, 92 Blephilia, 4 B. ciliata, 4 Bluegrass, 3-5 Bobwhite quail, 1-8 Boneset, 4 BORCHERS, ANTHONY L., 93 Borrelia burgdorferi, 47-52 B. sp. from Western Kentucky, 47— 53 in Peromyscus leucopus, 47-53 serological evidence for, 47-53 Bosmina longirostris, 101 Boyeria vinosa, 41, 43, 44 Broomsedge, 4 BROWN, G. C., 104 Buck Creek, 41-46 Bull Thistle, 4 BUMGARDNER, CLOYD J., 94, 95 BURKS, A., 80 Bush, strawberry, 4 Butterfly weed, 4 142 BYERS, M. E., 80, 81, 83, 87 BYRD, L. C., 84 Bythotrephes cederstroemii, 101, 102 Calopterygidae, 44 Calopteryx maculata, 41, 44 C. sp., 45 Campsis radicans, 4 Canada Thistle, 4 Canis latrans, 128 Capillaria didelphis, 130 C. longicauda, 130 C. spp., 128, 129 Carcinogenesis, mechanisms of in- tracellular niacin status, 90 Carex sp., 4 Carolina rose, 5 Carp, common, 23 Carpinus caroliniana, 110 CARTGER, J., 83 Carya ovata, 4 C. spp., 109, 110 Catonotus, 28, 29 Cedar, eastern red, 2 Celtis occidentalis, 4 Chaoborus, 103 Chemistry, college, 84 Cherry, black, 5 Chestnut oak, 5, 110 CHILD, MARY, 93 China, Hunan, 9-14 Chrysanthemum leucanthemum, 4 Cinquefoil, 5 Cirsium arvens, 4 C. vulgare, 4 CLARK, ROSS C., 134, 138 CLAUSON, JOHN M., 84 Clover, ladino, 3 red, 3,5 white, 5 white sweet, 4 yellow hop, 5 yellow sweet, 4 Coenagrionidae, 44, 45 Colinus virginianus, 1-8 Coilege chemistry, 84 prediction of successful students, 84 COMMENTS, 141 Common carp, 23 Composts, 83 for growing vegetables in contain- ers, 83 Computer-aided instruction in ge- ology, 85 CONRAD, PHILIP G., 85 Constructed wetlands, 81 influence on nutrient removal, 81 influence on pathogen removal, 81 treating livestock waste waters with, 82 Converted fescue field, 1-8 Coralberry, 5 Corduliidae, 43 Coreopsis lanceolata, 4 CORGAN, JAMES X., 54 Cornus florida, 109, 110 C. obliqua, 4 Cottontail rabbit, 1—S Cottontail rabbits, eastern, 128 Coyote, 128 Crabgrass, 4 Crataegus sp., 4 Croton sp., 4 Crownbeard, 5 Cruzia americana, 128, 129, 130 Cryptococcus neoformans, 84 serotype groups in, 54 CURRENS, JAMES C., 86 Cyclonaias tuberculata, 115 Cyprinus carpio, 23 Dacthal, persistence in soil, 80 Dactylis glomerata, 3-4 DAHLMAN, DOUGLAS L., 88 Daisy fleabane, 4 Daisy, ox-eye, 4 Damselfly community, 41-46 of Buck Creek, 41-46 of Pulaski County, Kentucky, 41— 46 Danthonia spicata, 4 Daphnia lumholtzi, 101-103 appearance of in the Ohio river, 101-103 impacts of in the Ohio river, 101— 103 Darter, duskytail, 28—40 striped fantail, 28-40 Daucus carota, 4 DAVIDSON, BART, 86 DEATON, E., 83 DEBSKI, ELIZABETH A., 90 Decomposition of mustard gas ana- logs using nicotine and ultra- sound, 15-21 Decomposition of nerve gas analogs using nicotine and ultrasound, 15—21 Deer tongue, 5 Delta-9 fatty acid desaturase, 91 organelle-specific role of, 91 Delta-aminolevulinic acid dehydra- tase, 87 in human erythrocytes, 87 Deptford pink, 4 Desmodium paniculatum, 4 Dewberry, 5 Dianthus armeria, 4 Didelphis virginiana, 128-133 Didymops transversa, 43, 44 Digitaria sanguinalis, 4 DINGER, JAMES S., 85 INDEX TO VOLUME 56 Dipsacus sylvestris, 4 Dirichlet, parts from, 87 Distinguished scientist awards, 96— 98 Dock, 5 DOCKSTADER, DAVID R., 85 Dogwood, 109 Dogwood, silky, 4 Dogwoods: molecules, morphology, and relationships, 84 DOTSON, O. W. III, 82 Doveweed, 4 DOWLA, H. A., 87 Dragonfly community, 41—46 of Buck Creek, 41-46 of Pulaski County, Kentucky, 41— 46 Dreisena polymorpha, 101 Dromogomphus spinosus, 43 Dropseed, prairie, 3, 5 DUOBINIS-GRAY, LEON, 47 Duskytail darter, 28-40 EAKIN, DAVID A., 134 Eastern cottontail rabbits, 128 Eastern Kentucky Coal Field, 84 hydrochemical facies in, 84 Eastern red cedar, 2 Echinococcus spp., 132 Edestus heinrichii, 54-56 Edestus minor, 54-56 Elkhorn Creek Monitoring Pro- gram, 80 ELLIOTT, LARRY P., 84 Elliptio dilatata, 114, 115, 116 ELLIS, AMY W,, 91 Elm, winged, 5 Elymus virginicus, 4 Enallagma basidens, 44 E. civile, 44 E. divagans, 44, 45 E. exculans, 44 Endothelial cells, 88 fatty acid effects, 88 vitamin E levels in, 88 Epitheca princeps, 43 Epizootic systems, comparing, 104— 108 using autoregression, 104-108 using spectral analysis, 104-108 Equine protozoal myeloencephalitis, 95 Eragrostis capillaris, 4 Erigeron annuus, 4 Erodible land, 83 vegetable culture on, 83 Erythemis simplicicollis, 43, 44 Escherichia coli, 90 Esfenvalearate, 80 analysis of on vegetables, 80 Etheostoma glabellare lineolatum, 28-40 E. kennicotti, 28, 29, 38, 39 E. percnurum, 28, 33-38 143 E. smithi, 39 E., species group, 25-40 Euonymus americanus, 4 Eupatorium serotinum, 4 Euphorbia, 4 E. corollata, 4 E. maculata, 4 F9 teratocarcinoma cells, 83 apoptosis in, 83 Fagus grandifolia, 110 Family size human sex ratios influ- encing, 9-14 Fantail darter, striped, 25-40 Fatty acid effects on vitamin E lev- els, 88 Fescue, converted field, 1-8 Fescue, tall, 3, 4 Festuca arundinacea, 1-8 Field goldenrod, 5 Fleabane, daisy, 4 Flowering spurge, 4 FORUM, 138-140 Foxtail, 5 Fraxinus americana, 4 FRAZIER, DR. DONALD T., 97 Frosty grape, 5 Gleditsia triacanthos, 4 Gnaphalium, 4 G. obtusifolium, 4 Goldenrod, field, 5 Goldenrod, gray, 5 Gomphidae, 43 Gomphus (Agrigomphus) lentulus, 43 G. (Gomphurus) lineatifrons, 43 G. (Hylogomphus) viridifrons, 43 Gongylonema longispiculum, 132 G. spp., 128, 131 Gragaria virginiana, 4 GRANSTROM, D. E., 95 Grape, frosty, 5 Grass, greasy, 5 love, 4 orchard, 4 panic, 5 poor man’s, 4 Gray goldenrod, 5 GRAY, ELMER, 9, 80, 81, 82, 83 Greasy grass, 5 Greenbriar, 5 GREENE, D., 80 Ground cherry, 5 Ground water, effect of fractures on, 85 Ground-water quality, 85 effect of agriculture chemicals on, 85 GUEORGUIEVA, D. S., 89 Hackberry, 4 Hagenius brevistylus, 43 HALEY, BOYD E., 91 HAMPSON, STEVEN K., 85 144 TRANS. KENTUCKY ACADEMY OF SCIENCE 56(3-4) HARPER, LEVI, 91 HARTMAN, D. R., 84 HASIBUAN, R., 104 Hataerina americana, 41 Hawthorn, 4 Helianthus mollis, 4 Heliothis virescens, 88 Hemlock, 110 Hemp, Indian, 4 HENNING, BERNARD, 88 Herbs, container grown, 81 Herbs, grown in composted mate- rials, 81 Hetaerina americana, 44 H. sp., 45 Hickory, 109 shagbark, 4 HIESTAND, DAVID M., 91 HILBORN, D., 80, 83 HILDEBRAND, DAVID F., 91 HINES, MARTINA, 109 Honey locust, 4 Honeysuckle, 4 HONG, IRENE, 92 Horse nettle, 5 HOUP, RONALD E., 114 Houstonia, 4 H. purpurea, 4 Human erythrocytes, 87 delta-aminolevulinic acid dehy- dratase in, 87 Human sex ratios, 9-14 in Hunan, China, 9-14 influencing family size, 9-14 Human T-Cells, 93 synthesis of insulin-like growth factor, 93 Hunan, China, 9-14 HUNT, ARTHUR, 89 HUNT, GRAHAM, 86 HURT, VALINA K., 9 Hydrochemical facies, 84 in Eastern Kentucky Coal Field, 84 Hydrostratigraphic units, 86 associated with carbonates, 86 associated with lake deposits, 86 associated with uranium-bearing black shales, 86 in Jefferson County, 86 HYLBERT, DAVID K., 86 Hypericum punctatum, 4 Index Herbariorum Kentuckiensis III, 134-137 Indian hemp, 4 Indian mock strawberry, 81 environmental effects on, 81 Inner Blue Grass Region, 85 occurrence of nitrate, 85 occurrence of pesticides, 85 Intracellular niacin status, 90 mechanisms of carcinogenesis, 90 Ipomoea pandurata, 4 Ironweed, 5 Ischnura posita, 44 I. verticalis, 44 Ivy, poison, 5 Ixodes pacificus, 47 I. scapularis, 47 JACK, JEFFREY D., 101 JACOBSON, ELAINE L., 90, 91 JETT, PAUL, 92 JOHNSON, RAY E., 82 JONES, LAKEASHA, 81 JONES, RONALD L., 134 Justicia americana, 44 KALISZ, PAUL J., 109 KEAGY, DWAYNE M., 85 Kentucky Groundwater Data Re- pository, 86 KERA, 94, 95 learner outcomes, 94 biomedical research/societal val- ues, 94 enhancing, 95 enhancing student perception of science, 95 in secondary science classrooms, 94 model classroom curriculum, 94 KESSLER, BRYAN, 82 Kleber Wildlife Management Area, 1-8 KOBRAEI, H. R., 93 Korean lespedeza, 3, 4 KORNMAN, LEWIS E., 57 KWANTES, MICHAEL G., 81 LACEFIELD, BRIAN D., 81, 82 Ladies tresses, 5 Ladino clover, 3 LAHAMER, AMER, 117 Lampsilis cardium, 115 L. fasciola, 115 L. ovata, 115 LANGLEY, CHRIS, 117 Lasmigona costata, 115 LAYMAN, STEVEN R., 28 LAYNE, DESMOND R., 81 Lespedeza procumbens, 4, 6 L. striata, 3, 4, 6 L. virginica, 4 Lespedeza, Korean, 3, 4 trailing, 4 Virginian, 4 Libellula, 42 L. luctuosa, 42, 43 L. lydia, 42, 43, 44 L. pulchella, 42, 43, 44 Libellulidae, 42, 43, 44 Lightning, 87 deaths and injuries from in Ken- tucky, 87 Ligumia recta, 115 Lipid peroxidation, 92 in broccoli florets, 89 influence of antioxident treat- ments, 89 influence of temperature treat- ments on, 89 Lireodendron tulipifera, 110 Livestock, 80 influence on water quality, 80 Locust, black, 5 Locust, honey, 4 LOH, WAI KHAN, 88 Lone star tick, 47 Longistriata didelphis, 128, 129, 130 Longistriata/Viannaia, 129, 130 Lonicera japonica, 4 Loosestrife, whorled, 4 Love grass, 4 Ludwigia sp., 4 LYONS, EUGENE T., 128 Lysimachia quadrifolia, 4 Maclura pomifera, 4 Macromia alleghaniensis, 43, 44 Macromiidae, 43, 44 Madtom, slender, 57 Magnolia sp., 110 Malathion, 15, 18-19 Manure application, 82 on corn, 82 plant analysis following, 82 soil analysis following, 82 Maple, red, 109 MCGRAW, LISA, 88 MCNEIL, RAYMOND C., 93 MEISENHEIMER, JOHN L., 15 Melilotus alba, 4 M. officinalis, 4, 5-6 Mephitis mephitis, 128 Methyl jasmonate, 89 influence of in broccoli, 89 influence on lipid peroxidation, 89 MIDDENDORF, BRAD, 88 Milkweed, big, 4 Millein, woolly, 5 MILLER, LAWRENCE R., 15 MILLER, RICHMOND J., 94 MIMS, STEVEN D., 22 Mite, twospotted spider, 104 Molineus barbatus, 128, 131, 132 Monarda, 4 M. fistulosa, 4 MORFORD, LORRI ANN, 93 Morning glory, 4 Mossbauer study, 117-127 of single crystal biotite, 117-127 of single crystal phlogopite, 117— 27 MUKHERJEE, R., 87 Multiflora rose, 5 MURRELL, ZACK E., 84 Muscrats, 128 Mussels, in Rockcastle River, 114— 116 Mustard, 4 Myeloencephalitis, equine proto- zoal, 95 Nematodes, 128-133 in Didelphis virginiana, 128-133 in the opossum, 128-133 Neozygites floridana, 104 Nettle, horse, 5 Neurocordulia yamaskanensis, 43 Neuronal death, 92 polyunsaturated fatty acids in, 92 NEWS, 99, 141 Non-protein amino acid disruption in proteins, 92 Nonpoint-source pollutant loads, 86 in a karst aquifer, 86 in an intensive-use agricultural re- gion, 86 NORDIN, G. L., 104 Noturus exilis, 57 Nyssa sylvatica, 109, 110 Oak, black, 109, 111 chestnut, 5, 110 post, 111 scarlet, 111 southern red, 109, 111 white, 109, 111 Obeliscoides cuniculi, 128, 131, 132 Obovaria subrotunda, 115 Odocoileus virginianus, 51 Odonata community, 41-46 of Buck Creek, 41-46 of Pulaski County, Kentucky, 41- 46 Ondatra zibethica, 128 Onion, wild, 4 Opossum, nematodes found in, 128-133 Orchard grass, 3, 4 Osage orange, 4 Osmorhiza sp., 4 OUTSTANDING TEACHER AWARDS, 96-98 Ox-eye daisy, 4 Oxalis stricta, 4 Oxydendrum arboreum, 109, 110 Pachydiplax, 42 P. longipennis, 43, 44 Paddlefish, oocyte staging in, 22-27 PANEMANGALORE, M., 87 Panic grass, 5 Panicum capillare, 5, 6 P. clandestinum, 5 P. microcarpon, 5 Parthenocissus quinquefolia, 5 Parts from Dirichlet, 87 Paspalum, 5 Paspalum sp., 5 Pawpaw, 81 growth and development of, 81 influence of fertilization on, 81 influence of incident light inten- sity on, 81 INDEX TO VOLUME 56 PAYNE, RANDALL G., 41 Pelurobema coccineum, 115, 116 Perca flavescens, 101 Perch, yellow, 101 Peromyscus gossypinus, 52 P. leucopus, 47-53 evidence for Borrelia sp. in 47-53 from Western Kentucky, 47-53 Petunia, wild, 5 Phlogopite, Méssbauer study of, L127. Physalis sp., 5 Physaloptera maxillaris, 128, 130, 131 P. spp., 128, 131, 132 P. turgida, 128, 129 Picoides borealis, 109-113 in Kentucky, 109-113 Pine, pitch, 111 Virginia, 111 Pink, Deptford, 4 Pinus echinata, 110 P. rigida, 110, 111 P. virginiana, 110, 111 Pitch pine, 111 Plantago, 5 Plantago lanceolata, 5 P. major, 5 Poa pratensis, 3, 5-6 Poison ivy, 5 Polyodon spathula, 22-2 oocyte staging in, 22-27 Poor man’s grass, 4 POPKEN, GREGORY J., 94 Poplar, yellow, 110 Post oak, 111 Potamilus alatus, 115 Potentilla simplex, 5 Prairie dropseed, 3, 5 Primary production, effects of nu- trients on, 93 effects of retention time on, 93 Procyon lotor, 51-52 PROGRAM, ANNUAL MEET- ING, 62-79 Protease nexin-1 in neural tissue, 92 relationship to Alzheimer’s Dis- ease, 92 Protozoal myeloencephalitis, equine, 95 Prunus serotina, 5 Ptychobranchus fasciolaris, 115 Pulaski County, Kentucky, 41-46 Pussy-toes, 4 Quadrula pustulosa, 114, 115, 116 Quail, bobwhite, 1-8 Queen Anne’s lace, 4 Quercus alba, 109, 110 Q. coccinea, 110, 111 Q. falcata, 109, 110, 111 Q. prinus, 5, 110 Q. stellata, 110, 111 Q. velutina, 109, 110, 111 145 Rabbit, cottontail, 1—8 eastern cottontail, 128 Ragweed, 4 RAMEY, DR. BARBARA A., 96 Raspberry, 5 Rattlebox, 4 Red cedar, eastern, 2 Red clover, 3, 5 Red maple, 109 Red oak, southern, 109, 111 Red-cockaded woodpeckers, in Kentucky, 109-113 Rhododendron, 110 Rhododendron maximum, 110 Rhus copallina, 5 R. glabra, 5 Robinia pseudoacacia, 5 Roof falls, geology of in under- ground coal mines, 86 Rosa carolina, 5 R. multiflora, 5 Rose, Carolina, 5 multiflora, 5 ROSENTHAL, G. A., 91, 92 ROSZMAN, THOMAS, 93 Rubus flagellaris, 5 R. pensilvanicus, 5 R. spp., 5, 6 Rudbeckia hirta, 5 Ruellia caroliniensis, 5 Rumex acetosella, 5 Russian sturgeon, 25 Rye, wild, 4 Sanicula, 5 Sanicula canadensis, 5 Sassafras albidum, 110 Scarlet oak, 111 SCHULZ, WILLIAM D., 15 SCHUSTER, GUENTER A., 41 SCOWBY, MICHAEL S., 91 Sedge, 4 Segregation plasmid F, in Escherich- ia coli, 90 SENDLEIN, LYLE V. A., 85 Senescence, in broccoli florets, 89 Setaria glauca, 5 Shagbark hickory, 4 Sharks, Pennsylvanian from Ken- tucky, 54-56 SHELTON, WILLIAM L., 22 SHIBER, JOHN G., 94 Sicily, sweet, 4 Silky dogwood, 4 SIMON, THOMAS P., 28 Skunks, striped, 128 Slender madtom, 57 Small Magellanic Cloud, B supergi- ants in, 93 MK classifications of, 93 Ultraviolet spectral morphology, 93 SMATHERS, KAREN L., 114 Smilax bona-nox, 5 146 TRANS SMITH, BETH, 89 Smooth sumac, 5 SNODDY, ED, 47 SNOW, MARY M.., 87 SNYDER, BRAD, 128 SNYDER, R., 83 Solanum carolinense, 5 SOLE, JEFFERY D., 1 Solidago altissima, 5, 6 S. nemoralis, 5 Somatochlora linearis, 41 Somatostatin-like immunoreactive cells in tadpole tectum, 90 Sourwood, 109 Southern red oak, 109, 111 Soybeans, biomass production in, 80 effects of cultivars on, 80 effects of planting dates on, 80 SPEAR, BRETT, 91 SPECK, R. R., 93 Spiranthes cernua, 5 Sporobolus asper, 3, 5-6 Spurge, flowering, 4 St. John’s wort, 4 STAMPER, SHELBY, 95, 128 STEINER, MARION, 92 Sticky purple stuff, 5 STILES, DAVID, 82 Strawberry bush, 4 Strawberry, mock, 81 wild, 4 Streptococcus equi subsp. zooepi- demicus, 92 surface protein variation of, 92 Striped fantail darter, 28-40 Striped skunks, 128 Strongyloides spp., 128, 129, 130, 131 Strophitus undulatus, 115 STUART, JAMES, 47 Sturgeon, Russian, 25 white, 22 Stylogomphus albistylus, 43 Sumac, smooth, 5 winged, 5 Sunflower, 4 Surface protein variation, 92 of Steptococcus equi subsp. z0oe- pidemicus, 92 SUSILO, F. X., 104 Sweet clover, white, 4 yellow, 4 Sweet sicily, 4 Sylvilagus floridanus, 1-8, 128 Symmorium reniforme, 55, 56 Sympetrum vicinum, 43, 44 Symphoricarpos orbiculata, 5, 6 Tall fescue, 3, 4 TAM, S. M., 89 TAWHID, ASLAM, 80, 82, 83 Teasel, 4 Teratocarcinoma cells, apoptosis in, 83 Teratocyte secretory products, 88 effects on protein synthesis, 88 in larval stages of tobacco bud- worm, 88 Tetranychus urticae, 104 Tetranychus-Neozygites 104-108 Thistle, bull, 4 Canada, 4 THORP, JAMES H., 101 Tick trefoil, 4 TIMONEY, JOHN, 92 TNF cytotoxicity by NAD, modula- tion of, 91 Tobacco vein mottling virus, inter- actions of, 89 TOLLIVER, SHARON C., 128 Toxic compounds, effect on human erythrocytes, 87 Toxicodendron radicans, 5 Trailing lespedeza, 4 Trefoil, tick, 4 Trichostrongylus calcaratus, 128, 131, 132 T. spp., 131 Trichuris leporis, 128, 131, 132 Tridens flavus, 5, 6 Trifolium pratense, 3, 5 T. procumbens, 5 T. repens, 3, 5 Trischostrongylus spp., 128 Tritigonia verrucosa, 115 Trumpet vine, 4 Tsuga canadensis, 110 TUNGATE, DR. FRED L., 97 mycosis, Ulmus alata, 5 Uncinaria stenocephala, 128, 131, 132 Underground coal mines, roof falls in, 86 Unionidae, 114-116 Unknown mint, 5 Vegetable culture, on erodible land, 83 VENUGOPAL, SANDHYA, 93 Verbascum thaspus, 5 Verbena, 5 Verbena simplex, 5 Verbesina occidentalis, 5 Veronia altissima, 5 Viannaia viannai, 128, 129, 130 Vibernum rifidulum, 5 V. prunifolium, 5 Villosa taeniata, 114, 115, 116 V. trabalis, 114, 115 . KENTUCKY ACADEMY OF SCIENCE 56(3—4) Vine, trumpet, 4 Viola sp., 5 Violet, 5 Virginia creeper, 5 Virginia pine, 111 Virginian lespedeza, 4 Vitamin E levels, fatty acid effects, 88 Vitis vulpina, 5 VOGELPOHL, S., 83 WALKER, JOHN, 92 WANG, ZIXING, 47 Water quality, influence of livestock on, 80 WATT, CHRISTOPHER, 90 Western Kentucky Coal Field, la- custrine deposits of, 85 Western Kentucky University, sus- tainable agronomic research, 82 Wetlands, constructed, 81 White ash, 4 White clover, 5 White oak, 109, 111 White sturgeon, 22 White sweet clover, 4 WHITE, BEVERLY LYNN, 96 WHITE, STEPHEN, 47 Whorled loosestrife, 4 Wild onion, 4 Wild petunia, 5 Wild rye, 4 Wild strawberry, 4 WILSON, MANDALA V., 91 Winged elm, 5 Winged sumac, 5 Wingstem, 4 Wood sorrel, 4 Woodpeckers, red-cockaded in Ken- tucky, 109-113 WOODWARD, JEROLD, 88 Woolly mullein, 5 Wort, St. John’s, 4 WRIGHT, TENA M., 81, 82 WU, YU Y., 9 WUNSCH, DAVID R., 84 WYATT, STEPHANIE L., 94, 95 Yarrow, 4 Yellow hop clover, 5 Yellow perch, 101 Yellow poplar, 110 Yellow sweet clover, 4 YOUNG, F. S. III, 80, 81 Zebra mussel, 101 ZGODA, MICHAEL A., 90 ZHUANG, H., 89 ZIMMER, STEPHEN, 92 Zygoptera, 41 ie ih i / Dh Aauks eG de Pathe HS ue re | by a Al 253 Is fi ier i ir A, MG f 1b 9 - ane & i iA, ae we See) via): ri - Instructions for Contributors Original papers based on research in any field of science will be considered for publication _ in the Transactions. Also, as the official publication of the Academy, news and announcements _ of interest to the membership will be included as received. _ Manuscripts may be submitted at any time to the Editor. Each manuscript will be reviewed by _ One or more persons prior to its acceptance for publication, and once accepted, an attempt will _be made to publish papers in the order of acceptance. Manuscripts should be typed double spaced throughout on good quality white paper 82 x 11 inches. NOTE: For format of feature articles and notes see Volume 43(3-4) 1982. 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He is also responsible for checking all lite erature cited to make certain that each article or book is cited correctly. Extensive alterations on the galley proofs are expensive and costs will be borne by the author. Reprints are to be CONTENTS Daphnia lumholtzi: appearance and likely impacts of an exotic cladoc- eran in the Ohio River. Jeffrey D. Jack and James H. Thorp ........... 101 Comparing epizootic systems using spectral analysis and autoregres- sion: a case study on Tetranychus—Neozygites mycosis. F. X. Susilo, ‘ G. L. Nordin, G. C. Brown, and R. Hasibuan ................00ccseeeceseeeees 104 Foraging of Red-Cockaded woodpeckers (Picoides borealis) in Ken- tucky. Martina Hines and Paul J. Kallisz ..............2cccceeeeeceeseneseeeeees 109 Extended monitoring of mussels (Bivalvia: Unionidae) in the Rockcastle River at Billows, Kentucky, an historical site. Ronald E. Houp and Karen L: Smathers). cscs ccc een ol es BUA so atabeoke spo es mee ateaee 114. Mossbauer study of single crystal biotite and phlogopite. Amer S. La- hamer. and’ Chris d. Langley oo. cn a caadebnoe eve eeaeesnenesteneae 117 Nematodes found in the opossum (Didelphis virginiana) and four other species of mammals in central Kentucky in 1991. Eugene T. Lyons, Sharon C. Tolliver, and Shelby Stamper ................2.20cceecececceecneeees 128 Index Herbariorum Kentuckiensis III. Ronald L. Jones, David A. Eakin, ; and, Ross Gs Clark (teccr ese enn cok agen gidassaceseoesuacessaeer ana veranasamaaee ae 134 FORUM Biodiversity and Kentucky’s heritage. Ross C. Clark ...................-.... 138 NEWS'‘AND COMMENTS iio7 i onc a ee Soe) fos wea care kee aead ch a tee | 141 . TRANSACTIONS OF THE KENTUCKY ACADEMY OF SCIENCE 7, —_S” \WTHSON Ap MAY 3 1996 Volume 57 SOKARIES Number 1 March 1996 iy it J hs Official Publication of the Academy The Kentucky Academy of Science Founded 8 May 1914 GOVERNING BoarD FoR 1996 ExecutivE COMMITTEE President: William S. Bryant, Department of Biology, Thomas More College, Crestview Hills, KY 41017 President Elect: Marcus T. McEllistrem, Department of Physics, University of Kentucky, Lexington, KY 40506-0055 Vice President: Patricia K. Doolin, Ashland Petroleum Company, P.O. Box 391, Ashland, KY 41114 Past President: Robert O. Creek, Department of Biological Sciences, Eastern Kentucky University, Rich- mond, KY 40475 Secretary: Peter X. Armendarez, Department of Chemistry and Physics, Brescia College, Owensboro, KY 42301 . Treasurer: Julia H. Carter, Wood Hudson Cancer Research Laboratory, 931 Isabella Street, Newport, KY 41071 Executive Secretary (ex officio): J. G. Rodriguez, Department of Entomology, University of Kentucky, Lexington, KY 40546-0091 Editor, TRANSACTIONS (ex officio): John W. Thieret, Department of Biological Sciences, Northern Ken- tucky University, Highland Heights, KY 41099 MEMBERS, GOVERNING BOARD Patricia K. Doolin 1996 dames F. Hopgood 1998 David E. Hogan 1996 Robert H. Wombles 1998 Gerald L. DeMoss 1997 AAAS/NAAS Representative Wimberly C. Royster 1997 J. G. Rodriguez 1998 Chairperson, KJAS Vincent DiNoto 1996 COMMITTEE ON PUBLICATIONS Editor and John W. Thieret, Department of Biological Sciences, Northern Kentucky University, Chairman: Highland Heights, KY 41099 Associate Editor: James O. Luken, Department of Biological Sciences, Northern Kentucky University, Highland Heights, KY 41099 Abstract Editor: Robert F. C. Naczi, Department of Biological Sciences, Northern Kentucky University, Highland Heights, KY 41099 Index Editor: Varley Wiedeman, Department of Biology, University of Louisville, Louisville, KY 40292 Editorial Board: Vincent DiNoto, Natural Science Division, Jefferson Community College, Louisville, KY 40201 Jerry N. Baskins, School of Biological Sciences, University of Kentucky, Lexington, KY 40506 John P. Harley, Department of Biological Sciences, Eastern Kentucky University, Richmond, KY 40475 Robert O. 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Correspondence concerning memberships or subscriptions should be addressed to the Secretary. Exchanges and corre- spondence relating to exchanges should be addressed to the Librarian, University of Louisville, Louisville, KY 40292, the exchange agent for the Academy. This paper meets the requirements of ANSI/NISO Z39.48-1992 (Permanence of Paper). SSeS er rs > gic a INSTITUTIONAL AFFILIATES Fellow University of Kentucky University of Louisville Sustaining Member Eastern Kentucky University Northern Kentucky University Morehead State University Western Kentucky University Murray State University Member Berea College Hazard Community College Campbellsville College Paducah Community College Centre College Southeast Community College Cumberland College Thomas More College Associate Member Ashland Community College Maysville Community College Brescia College Midway College Georgetown College Somerset Community College Jefferson Community College — Spalding University Kentucky State University Transylvania University Kentucky Wesleyan College INDUSTRIAL AFFILIATES Associate Patron Ashland Oil, Inc. Sustaining Member Fruit of the Loom Member Alltech Biotechnology Center Corhart Refractories Corporation Martin Marietta, Inc. MPD, Inc. United Catalyst, Inc. Westvaco Associate Member 3M Tape Manufacturing Division, Cynthiana Plant All-Rite Pest Control Wood Hudson Cancer Research Laboratory, Inc. TRANSACTIONS of the KENTUCKY ACADEMY of SCIENCE March 1996 Volume 57 Number 1 Trans. Ky. Acad. Sci. 57(1):1—5. 1996. Intersexual Differences in Feeding Frequency and Prey Size in the Robber Fly Promachus albifacies (Diptera: Asilidae): Possible Influence of Male Mating Behavior Brian A. Kunkel and Jon M. Hastings Department of Biological Sciences, Northern Kentucky University, Highland Heights, Kentucky 41099-0400 ABSTRACT The robber fly Promachus albifacies (Williston) is an ambush aerial predator that feeds on a wide variety of insects. Late in the flight season, males spend most of the daytime on or near shrubs in which females oviposit. Males use the oviposition shrubs as mate encounter sites. We studied a population of P. albifacies in a desert grassland habitat in central New Mexico, testing the hypothesis that male occupation of oviposition shrubs constrains foraging. Results did not support the hypothesis. Late in the flight season, females did feed more frequently and on larger prey than did males. However, we observed no difference in feeding frequency or in prey size for males between early and late season, suggesting that intersexual differences in feeding can be attributed to factors other than constraints imposed by male mating behavior. INTRODUCTION Many field studies of robber flies described foraging behavior and cataloged the taxon and size of their insect prey. Several of these stud- ies compared feeding frequencies and prey size of male with female flies; the patterns vary among species. In certain species, females feed more frequently than males (Lavigne 1979; Weeks and Hespenheide 1985); in oth- ers no intersexual difference in feeding fre- quencies has been observed. In some species, females capture larger prey (Scarbrough 1979); in others, males capture larger prey (Lavigne 1979): and in others no intersexual difference in prey size is reported (Weeks and Hespenheide 1985). While most of these stud- ies provided excellent documentation of asilid foraging, few offered explanations for ob- served intersexual differences (exceptions: Dennis 1979; Scarbrough 1979), and none proposed and tested hypotheses attempting to account for the differences. In many species of insects sexual selection has led to evolution of behaviors in males that enhance mating success relative to other males (Thornhill and Alcock 1983). Behavior evolved in the context of competition for mates may be in conflict with behavior that enhances sur- vival (Gwynne 1987), including, conceivably, foraging behavior. Late in the flight season male Promachus albifacies spend most of the daytime perched on or near shrubs in which females oviposit (Hastings et al. 1994). Males use these shrubs as mate encounter sites. However, these sites appear to be inferior foraging sites (Hastings et al. 1994). We therefore hypothesized that male occupation of these sites constrains their foraging. During the late season, males should feed less often and on lower quality (i.e., smaller) prey than do females. During the ear- ly season, males should feed more often and on larger prey than during the late season, as Y Transactions of the Kentucky Academy of Science 57(1) males are less constrained by mating behavior during the early season. MATERIALS AND METHODS We studied a population of P. albifacies in central New Mexico between 20 May and 19 June 1993. The 18.4-ha study site is located on an alluvial fan at the north end of the Mag- dalena Mountains ca. 10 km east of Magda- lena, Socorro County. The habitat is a decent grassland containing sparse staghorn cholla (Opuntia arborescens) and narrow leaf yucca (Yucca elata) within a matrix of short grasses, a variety of short annual plants, and bare soil. Censusing Procedures Female P. albifacies oviposit in cracks and small openings in woody tissues of dead yucca flowering stalks and dead cholla branches that are at least 1 m above the ground (Hastings et al. 1994). We term cholla and yucca plants containing such sites as “oviposition shrubs.” We subdivided the study site into two areas: OS—oviposition shrubs and all land area with- in 1 m of an oviposition shrub, and OTHER— all area not within 1 m of an oviposition shrub. We counted the number of oviposition shrubs within the study site, estimated the average diameter of shrubs to be 1 m, and then cal- culated the percentage of the study site cov- ered by the two area subdivisions to be 1.2% OS, 98.8% OTHER. In 1993 the flight season of P. albifacies ex- tended from 20 May to at least 25 June at the study site; few adults were alive on or later than 25 June. To determine daily activity and spatial distribution of the robber flies we con- ducted censuses each day from 26 to 31 May and from 4 to 19 June. Censuses conducted from 26 to 31 May are defined as early-season censuses; those conducted from 4 to 19 June are defined as late-season censuses. Censusing effort each day was constant between 0900 and 1600 MDT. During censusing we walked along a grid of lines parallel to a border of the study site and used visual and auditory cues to locate flies. Grid lines were spaced 20 m apart to minimize repeated observations of the same individuals. We recorded the location (OS vs OTHER) and the activity (perch, feed, mate, oviposit) of each fly. We defined feeding as a fly having a prey item in its possession. A fly was considered to be perching if it was not engaged in any of the other activities. Prey Size and Prey Taxon We captured samples of feeding male and female P. albifacies during the early and late season. Dial calipers were used to measure body length of their insect prey to the nearest 0.05 mm. We grouped prey into size classes of 5 mm increments, and used Kolmogorov- Smirnov tests to compare the size-class distri- butions of prey captured by male vs female robber flies, and by males in the early vs the late season. We identified the order of each insect prey item retrieved from the robber flies. Intersexual Body Size Comparison To estimate body size of flies in the field, we used dial calipers to measure the length of the right wing to the nearest 0.05 mm. Wing length and dry body mass are highly correlated in both sexes (Hastings et al. 1994). We mea- sured the wing length of a random sample of male and female P- albifacies and used a stu- dent's t-test to compare the sample means. RESULTS As Hastings et al. (1994) previously report- ed, the activity of female P. albifacies varied between seasons (Chi-square = 28.9, P < 0.0001, N = 536) (Fig. 1). Ovipositioning fre- quency was higher during the late season (57 of 259 sightings of females) than it was during the early season (23 of 277 sightings). Also as Hastings et al. (1994) reported, we found that late season male spatial distribution was non- random (Chi-square = 569.7, P < 0.0001, N = 735). Despite the fact that OS area covered only 1.2% of the study site, 427 of the 735 sightings of males were in OS locations. Males were Also nonrandomly distributed within the study site during the early season (Chi-square = lid. 2 = 000015 N — 602) s butemales were found less frequently in OS locations than they were in the late season (Chi-square = 180) 45 1? = OOOO IN = 1887) (Fig. om Though P. albifacies males and females feed on a wide variety of insects (Table 1), they feed largely on insects that forage on flowers; of the 134 Hymenoptera prey captured by males and females combined, 121 were bees (Apidae); of the 119 Diptera prey, 80 were bee 60 E ; || early late 50 | 40 | % Frequency wo oO oe oe Feed Perch Mate Oviposit Activity 60 _| females ial males 50 40 7 30 % Frequency 20 | if 20. .725 30 Prey size (mm) 5 10 15 Figures 1-4. different activities during the e arly (N = 277 Male Mating Behavior in Promachus albifacies—Kunkel and Hastings The robber fly Promachus Bale ies. Fig. | ) and during the late flight season (N = 259). Fig. 2 (upper right). ey) 80 | = early | | late 60 40 | ‘% Frequency 20 | os OTHER Location 50 a males females 40 30 20 % Frequency 1 ie) 15) Ws a7 ate! IS) 20) Pal 22 Wing length (mm) io) Percent Been: ney of females engaged in (upper left). Percent frequency of males sighted in OS ( (oviposition shrubs) vs OTHER locations within che habitat during e arly season (N = 602) and late season (N = 735) censuses. Fig. 3 of prey of males (N = 54) vs females (N = 31) of size (wing length) classes of males During the late season, fe- frequency than did males flies (Bombyliidae). males fed at a higher { (Chi-square = 5. 3, P= the 735 males sighted, 63 had prey in their possession; 36 of 259 females were observed (lower left). captured during the late season. Fig. 4 (N = 78) vs females (N = 76) 0.02, N = 994). Of (total body le ngth) classes Percent frequency of size Percent frequency (lower right). with prey. Prey size also varied between sexes during the late season (Kolmogorov-Smirnov = 0.56, P < 0.0001, male N = 54, female N = 31) (Fig. 3); females generally captured larger prey. 4 Transactions of the Kentucky Academy of Science 57(1) TaBLE 1. Number and percent composition of prey by order taken by the robber fly Promachus albifacies for 1992 and 1993 in central New Mexico. Males Females Total Order Number Percent Number Percent Number Percent Coleoptera 84 32.1 25 13.0 109 24.0 Diptera 55 =21.0 64 33.3 119 26.2 Hemiptera ial O:3 21 10.9 48 10.6 Homoptera 4 1.5 10 5.2 14 3.1 Hymenoptera 78 29.8 56 29.2 134 29.5 Lepidoptera 9 34 3 1.6 26 Neuroptera 2 0.8 3 1.6 5 1.1 Orthoptera 3 L.1 10 5.2 13 2.9 Totals 262 192 454 Male feeding frequency did not vary signif- icantly between seasons (Chi-square = 0.99, P = 0.32, N = 1337); of 602 late-season sight- ings of males, 63 had prey; 65 of 735 males censused during the early season had prey. Size of prey captured by males also did not vary significantly between seasons (Kolmogo- rov-Smirnov = 0.08, P = 0.09, late N = 54, early N = 60). The mean (+ SEM) wing length of a ran- dom sample of 76 female P. albifacies (18.05 + 0.30 mm) was larger than the mean wing length of a random sample of 78 males (16.30 + 0.20 mm; t = 6.9473, P < 0.0001) (Fig. 4). DISCUSSION We observed the same basic seasonal changes in activity of male and female P. al- bifacies that Hastings et al. (1994) reported. Ovipositioning frequency within the female population was higher in the late than in the early flight season. In the late season males spent more of the daytime within the relative- ly sparse OS locations than they spent at these sites early in the season when visits to them by females were relatively infrequent. Male perch selection, at least during the late season, appears to be largely driven by competition for mates. Hastings and others (1994) reported that the OS locations were inferior foraging sites. This is consistent with our finding that P. al- bifacies of both sexes feed to a large extent on pollinating insects. During the flight season many plants within the habitat are in bloom, but the oviposition shrubs, which consist of dead yucca flower stalks or partly to complete- ly dead cholla, are generally devoid of flowers. Consequently, fewer preferred prey should be available in oviposition shrubs than in other locations. Therefore, we expected that male occupation of the shrubs would reduce the number and/or quality of prey available to them. However, our prediction that the in- creased male occupation of ovipositioning shrubs during the late season would constrain foraging was not supported by the results. The observation that male feeding frequency and prey quality did not differ between seasons suggests that male mating behavior does not constrain foraging. If male mating behavior does not impose constraints on foraging, then what accounts for the observed intersexual differences in feeding frequency and prey size? In general, the eggs of female robbers flies are not mature when they eclose, and ovipositioning is de- layed 10 to 11 days following emergence (La- vigne et al. 1978). Vitellogenesis, which con- tinues throughout the adult life of females (Scarbrough 1978) increases the nutritional demands of females over those of males (Scar- brough 1979); females simply must feed more often and/or on larger prey. Shelly (1985) re- ported that, in general, larger robber flies cap- ture and feed on larger prey. The fact that female P. albifacies are generally larger than males probably enables them to capture and feed on larger prey. SUMMARY During the late flight season, male P. albi- facies spend most of the daytime on shrubs in which females oviposit. The shrubs, which males use as mate encounter sites, have pre- viously been reported to be inferior foraging locations. We tested the hypothesis that male occupation of these shrubs would constrain their foraging. Results did not support the hy- pothesis. Though during the late season, fe- males fed more often and on larger prey than did males, male foraging frequency and prey size did not vary between the early and the late season. This suggests that intersexual dif- ferences in foraging during the late season re- sult from factors other than constraints im- posed by male mating behavior. We suggest that the nutritional demands of vitellogenesis and the fact that females are generally larger than males may account for the observed in- Male Mating Behavior in Promachus albifacies tersexual differences in foraging frequency and prey size. LITERATURE CITED Dennis, D. S. 1979. Ethology of Holcocephala fusca in Virginia (Diptera: Asilidae). Proc. Entomol. Soc. Wash. 81:366-378. Gwynne, D. T. 1987. Sex-biased predation and the risky mate-locating behaviour of male tick-tock cicadas (Ho- moptera: Cicadidae). Anim. Behav. 35:571-576. Hastincs, J. M., G.N. Dopson, and J. L. Heckman. 1994. Male perch selection and the mating system of the robber fly, Promachus albifacies (Diptera: Asilidae). J. Insect Behav. 7:829-841. LavicnE, R. J. 1979. Notes on the ethology of Efferia argyrogaster (Diptera: Asilidae) in Mexico. Proc. En- tomol. Soc. Wash. 81:544—551. LavicnE, R., S. DENNIS, and J. A. Gowen. 1978. Asilid Kunkel and Hastings 5 literature update 1956-1976 including a brief review of robber fly biology (Diptera: Asilidae). Agricultural Ex- periment Station, University of Wyoming, Laramie. ScarBRouGH, A. G. 1978. Ethology of Cerotainia albi- pilosa Curran (Diptera: Asilidae) in Maryland: court- ship, mating and oviposition. Proc. Entomol. Soc. Wash. 80:179-190. . 1979. Predatory behavior and prey of Diogmites missouriensis Bromley in Arkansas (Diptera: Asilidae). Proc. Entomol. Soc. Wash. 81:391—400. SHELLY, T. E. 1985. Ecological comparisons of robber fly species (Diptera: Asilidae) coexisting in a neotropical forest. Oecologia 67:57—70. THORNHILL, R., and J. ALcock. 1983. The evolution of insect mating systems. Harvard University Press, Cam- bridge, MA. Weeks, L., and H. A. HesPENHEIDE. 1985. Predatory and mating behavior of Stichopogon (Diptera: Asilidae) in Arizona. Pan-Pac. Entomol. 61:95—104. Trans. Ky. Acad. Sci. 57(1):6-14. 1996. Effects of Wastewater Discharge from Houseboats on Water Quality in a Large Oligotrophic Reservoir, Lake Cumberland, Kentucky David Loy and Paul A. Bukaveckas!' Water Resources Laboratory, University of Louisville, Louisville, KY 40292 ABSTRACT Effective water quality management depends on availability of site-specific data on nutrient inputs, outputs, and transport. We examined the potential importance of wastewater discharge from houseboats as a source of nutrient and organic matter loading to a large, oligotrophic reservoir (Lake Cumberland, Kentucky). Spatial variation in chlorophyll, nutrient, and dissolved oxygen concentrations were compared between areas differing in levels of houseboat activity. Upstream-downstream gradients in chlorophyll, transparency, and nutrient availability generally followed expected patterns based on longitudinal variations in flow velocity and water residence time. Consistently elevated total phosphorus (TP) levels were observed immediately down- stream from a large houseboat marina complex. Although a corresponding increase in chlorophyll was not observed, higher rates of nitrate and silica depletion at this site suggested greater phytoplankton production. Overall, the data provided no compelling evidence of water quality deterioration in areas frequented by houseboats. The magnitude of nutrient and organic matter loading from direct wastewater inputs appeared to be small in relation to lake volume. INTRODUCTION Nutrient loading to surface waters is a wide- spread problem contributing to water quality deterioration in lakes, rivers, and estuaries. Nutrient inputs arise from agricultural runoff, industrial discharge, and municipal and do- mestic wastewater. Symptoms of excess nutri- ent loading include proliferation of certain kinds of algae and associated reductions in wa- ter clarity. Algal blooms and subsequent mi- crobial decomposition promote high biological oxygen demand, which can lead to periodic fish kills. Although effects of eutrophication have been well documented through monitor- ing and experimental approaches (e.g., Ed- mondson 1972; Schindler 1974), site-specific information on nutrient sources and tr ansport are often lacking. This limits the effectiveness of lake and watershed management plans for reducing nutrient inputs. We examined one potential source of nutrient inputs (wastewater discharge from houseboats) to a large reser- voir where perceived changes in water quality have been of recent concern. Lake Cumberland, a large (20,336 ha), oli- gotrophic reservoir in south-central Kentucky (Figure 1), was formed upon completion of Wolf Creek Dam on the Cumberland River in 1952. Its purpose was to provide hydroelectric power to parts of southern Kentucky and Ten- ' Author to whom correspondence should be addressed. nessee. In the last 20 years a thriving recrea- tion industry has developed with tourists from nearby metropolitan centers (Louisville, Cin- cinnati, and Indianapolis) visiting the lake and providing income for resorts and marinas around the reservoir. The growing tourism in- dustry is one factor that may pose a threat to water quality conditions in the lake. Particu- larly controversial has been the increase in numbers of houseboats (more than 500 in peak season) using the lake. Federal guide- lines require that boats be outfitted with Ma- rine Sanitation Devices to provide primary treatment of wastewater. Boaters are not re- quired to hold chemically treated wastewater for subsequent processing at an on-shore sec- ondary treatment facility. This has led to the common practice of flushing chemically treat- ed wastewater directly into the lake. In our study we examined spatial variations in water quality in relation to levels of house- boat activity. Wolf Creek embayment is an area frequently used by houseboats; Indian Hills Resort Marina and Alligator Dock are lo- cated along this embayment. Caney Creek, the other studied embayment, has no houseboat rental facilities and consequently has lower levels of houseboat activity. Movement of houseboats between the embayments is lim- ited due to lack of adequate mooring sites in the Caney Creek embayment and to the long travel time resulting in high operational costs. Water Quality in Lake Cumberland—Loy and Bukaveckas Outline map of Kentucky, showing location of Figure 1. Lake Cumberland. We compared water quality conditions in Wolf Creek and Caney Creek embayments as an in- dex of effects of houseboat activity. Measured parameters included algal abundance (as chlo- rophyll a), nutrient concentrations (total phos- phorous, inorganic nitrogen, and dissolved sil- ica), and bacterial activity (rates of hypolim- netic oxygen depletion). We hypothesized that nutrient and organic matter loading associated with houseboat discharge would result in ele- vated nutrient concentrations, increased algal growth, and increased bacterial activity. We predicted that chlorophyll concentrations, nu- trient availability, and rates of oxygen deple- tion would be greater at the site experiencing more intense houseboat activity (Wolf Creek embayment). SITE DESCRIPTION AND SAMPLING DESIGN Lake Cumberland has a surface area of 20,336 ha, a mean depth of 24.2 m, and a max- imum depth of 57 m; it exhibits strong thermal stratification from April through October. The lake’s watershed (1,479,170 ha) is a mosaic of land-use types including 21% agricultural, 55% forestlands, and 3% urban areas (DEP 1984). Topography of the region is mountain- ous in the east, leveling to gentle plateau in the west; deep narrow valleys etched into the landscape form the reservoir’s basin. Major in- flows include the Cumberland River and its Big South Fork; minor contributors are nu- merous creeks throughout the length of the reservoir. The sole major outflow is the re- maining Cumberland River below Wolf Creek Dam. To characterize spatial variation in each of the embayments, we sampled three stations along a transect from the main channel of the lake toward the headwaters (Figure 2). The three Caney Creek stations and the furthest — ( Caney Creek F Alligator Wolf Creek olf Creek C-u P W-up C-mid—"“A W-mid N W-dn Area Enlarged AS ie AH Figure 2. Map of Lake Cumberland, Kentucky, showing location of sampling sites in Wolf Creek embayment (W- up, W-mid, and W-dn), Caney Creek embayment (C-up, C-mid, and C-dn), and open lake (OL). downstream Wolf Creek site (W-dn) are in un- developed areas. The upstream and mid-em- bayment Wolf Creek stations (W- up and W- mid, respectively) are close to marinas with associated par king and recreation areas. Sta- tion W-mid is 100 m downstream from Indian Hill Resort Marina and directly across from a small shale beach on the west bank heavily used for swimming and recreation. Station W- up is 200 m upstream from Alligator Dock. Lake depth varies among sampling sites: from 10 m at the upstream stations to >25 m at sites further downstream. Samples were collected also at an open-lake (main channel) site to assess differences in wa- ter quality relative to the embayments. The sampling site in the main body of the lake (at Cumberland River mile 469.7) was chosen be- cause it is a U.S. Army Corps of Engineers water quality monitoring site (samples collect- ed twice yearly, in mid-summer and early fall). The site is exposed to prevailing winds and, as a result, displays much deeper mixing patterns than the embayments. Water level in the res- ervoir varies considerably, decreasing from maximum pool in spring to minimum pool in 8 Transactions of the Kentucky Academy of Science 57(1) late summer. Depth at the open-lake site var- ies from 45 m in March to 38 m in October. Samples were collected at monthly intervals between 1 May and 1 Oct 1994. To charac- terize water quality conditions while the lake was at maximum pool, one set of samples was collected in mid-March (Wolf Creek sites only). METHODS Temperature and irradiance profiles were obtained for each of the seven sites from mea- surements taken at 1 m intervals (surface to 25 m or to bottom at shallower sites). Tem- perature was measured with a YSI Model S- C-T thermistor. Irradiance profiles (PAR) were measured with a Protomatic photometer equipped with upward and downward spher- ical sensors. Depth profiles of downwelling and upwelling irradiance were analyzed using the methods of Kirk (1983) to estimate coef- ficients of light attenuation, absorption, and scattering. The attenuation coefficient for downwelling irradiance (Kd) was determined from a linear regression of the natural log of downwelling irradiance against depth. Corre- lation coefficients derived by fitting linear re- gressions to irradiance data were uniformly high, and estimates for the standard error of the slope were consistently less than 10%. A detailed description of analytical procedures used for irradiance data is presented in Bu- kaveckas and Driscoll (1991). Water samples for chlorophyll analyses were taken at three depths spaced equally between the surface and the 1% light level. Samples were stored in coolers and processed, within 1 to 3 hours after collection, by filtration through Gelman A/E glass fiber filters. Filters were stored frozen and in the dark. Within 3 to 10 days the filters were macerated and pig- ments extracted overnight in 90% acetone. Extracts were analyzed using a Varian DMS 70 spectrophotometer equipped with long pathlength (4 cm) cells and narrow (1 nm) bandwidth. Concentrations of degraded chlo- rophyll a were corrected for pheophytin a us- ing the Lorenzen equations as modified by Speziale et al. (1984). Water samples for dissolved oxygen deter- minations were collected at 5 m intervals to 20 m (or to the bottom). Samples were ana- lyzed using the azide modification of the Winkler titration method (APHA 1985). Water samples for nutrient analyses were taken at depths of 3 m, 10 m, and 17 m (where pos- sible). All nitrate and dissolved silica analyses were performed on an autoanalyzer (Skalar San Plus) using unfiltered water samples. Ni- trate concentrations were determined using the automated cadmium reduction method; dissolved silica was measured by the automat- ed molybdosilicate method (APHA 1985). To- tal phosphorous was analyzed on unfiltered samples using the manual two reagent ascor- bic acid method following persulfate digestion (APHA 1985). RESULTS Spatial variation in water column transpar- ency followed a consistent trend of increasing light penetration with increasing distance downstream (Figure 3). Attenuation coeffi- cients (Kd) ranged from 0.40 to 0.75 (summer averages) with highest light attenuation mea- sured at upstream sites (W-up and C-up). Variation in light attenuation corresponded to differences in depth of the photic zone (1% light level), which ranged from 7 to 8 m at upstream sites to 10 to 12 m at downstream and open-lake sites. At all stations, vertical gradients of chlorophyll showed a distinct peak at depths ranging from 5 to 10 m. These depths corresponded to light levels approxi- mating 2% to 10% of subsurface irradiance. Differences in light attenuation among sites located in the Caney Creek embayment were related to variation in light scattering. The im- portance of scattering in regulating overall light attenuation was inferred from the higher values measured upstream (coefficient = 2.0) and the lower values occurring downstream (coefficient = 0.5). By comparison, coeffi- cients of light absorption were smaller (0.4— 0.5) and exhibited little variation along the up- stream-downstream gradient. In the Wolf Creek embayment, variation in both scattering and absorption contributed to between-site differences in light attenuation. Light attenu- ation, highest at the furthest upstream site, decreased downstream despite higher scatter- ing at W-mid. Depth profiles of dissolved oxygen differed markedly between the embayments. Data col- lected on 6 July typify the differences that were apparent throughout the summer (Fig- Water Quality in Lake Cumberland—Loy and Bukaveckas 9 Light Attenuation i= aS = o 3 bed o ° ° C-up C-mid C-dn W-upW-midW-dn = OL Sampling Location Light Absorption i= ay = o Ss - o ° ° C-up C-mid C-dn W-upW-midW-dn OL Sampling Location 320) Light Scattering 225 = 2.0 x c 2 4S 9 5 ‘oO 1-0 ° 0.5 0.0 EB ES C-up C-mid C-dn W-upW-midW-dn- OL Sampling Location Figure 3. Spatial variation in coefficients of light atten- uation, light absorption, and light scattering along an up- __ stream-downstream gradient in two embayments and the main channel of Lake Cumberland, Kentucky (mean val- ues for four sampling dates in July—October 1994). ure 4). At all stations, upper water column samples (0—5 m) were at or above saturation (90% to 110%). Differences between the em- bayments were evident in samples collected at 10 m. At Caney Creek sites, 10 m samples were consistently within 10% to 20% of at- mospheric equilibrium (saturation >80%), 125 Dissolved Oxygen 100 75 50 Percent Saturation 25 5 1015 5 1015 5 1015 C-up C-mid C-dn Sampling Depth (m) and Site (Caney Creek) Dissolved Oxygen Percent Saturation 5 1015 5 W-up Sampling Depth (m) and Site (Wolf Creek) 10 15 5 W-mid 10 15 W-dn Figure 4. Oxygen saturation at 5, 10, and 15 m along an upstream-downstream gradient in the Wolf Creek and Ca- ney Creek embayments Lake Cumberland, Kentucky, for samples collected on 6 July 1994. B denotes missing sam- ples where lake depth <15 m. whereas 10-m samples from Wolf Creek sites were substantially depleted in dissolved oxy- gen (10% to 50% saturation). Differences in oxygen resources of the lower water column were most pronounced at upstream and mid- stream sites, whereas oxygen profiles obtained at downstream sites were generally similar. Patterns of spatial variation in chlorophyll and nutrient concentrations were similar be- tween embayments (Figure 5) with highest chlorophyll concentrations observed at sites furthest upstream. Chlorophyll concentrations upstream (C-up and W-up) typically exceeded 5 pg/liter (photic zone average), whereas val- ues for open-lake and midstream and down- 10 Transactions of the Kentucky Academy of Science 57(1) Chlorophyll ugxliter™ C-up C-mid C-dn W-up W-mid W-dn OL Sampling Location Total Phosphorus ug~liter* C-up C-mid C-dn W-up W-mid W-dn OL Sampling Location 60 Nitrate 50 40 30 ugxliter™ 20 10 C-up C-mid C-dn W-up W-mid W-dn OL Sampling Location Dissolved Silica mg«liter™ C-up C-mid C-dn W-up W-mid W-dn OL Sampling Location Figure 5. Spatial gradients in chlorophyll, total phosphorus, nitrate, and dissolved silica concentrations along an upstream- downstream gradient in two embayments and the main channel of Lake Cumberland, Kentucky. Data shown are mean values for samples collected between 0 and 10 m on four sampling dates in July—October 1994 (n = 30). stream sites were generally lower. Spatial vari- ation in TP followed patterns similar to those for chlorophyll. In both embayments, higher phosphorus concentrations were measured at upstream sites (15-20 yg/liter) with lower val- ues (<10 pg/liter) at downstream sites. The embayments differed in that phosphorus con- centrations were consistently higher at the midstream site in Wolf Creek (nearest to the marina) in comparison to the midstream site at Caney Creek. Patterns of spatial variation for nitrate dif- fered from those observed for chlorophyll and total phosphorus. Nitrate concentrations at upstream and midstream sites were extremely low (<10 g/liter) and in a few cases below analytical detection limits. Somewhat higher values were observed at downstream and open-lake sites (30-50 g/liter). Spatial pat- terns in dissolved silica were similar between embayments, with highest concentrations measured at downstream sites (>2 mg/liter) and lower concentrations upstream (1-2 mg/ liter). Seasonal variation in chlorophyll and nutri- ent concentrations were similar between em- bayments (Figure 6) although interpretation of these trends is somewhat compromised by the incomplete record for Caney Creek sites (no March data). Chlorophyll concentrations were notably higher at W-up and W-mid sites during the March sampling. Concentrations were generally low at all sites during summer (photic zone averages <10 g/liter). Nutrient concentrations in surface waters were also highest in March and declined throughout the summer. In the Wolf Creek embayment, total phosphorous decreased from a range of 50-75 pe/liter in March to 10-20 pg/liter by early July. Nitrate was depleted more rapidly with concentrations decreasing from 400-600 g/ liter to <5 pg/liter by July. The relative avail- ability of nitrogen to phosphorus was consis- tently low at all stations and throughout the Water Quality in Lake ¢ Chlorophyll rr W-up 2 W-mid = C-dn a W-dn Mar Apr May Jun Jul Aug Sep Oct Total Phosphorus s = x D St Mar Apr May Jun Jul Aug Sep Oct Figure 6. Jumberland Loy and Bukaveckas 1] 700 Nitrate 600 500 400 ugxliter™ 300 200 100 Mar Apr May Jun Jul Aug Sep Oct Dissolved Silica mgxliter™ Mar Apr May Jun Jul Aug Sep Oct Temporal trends in chlorophyll, total phosphorus, nitrate, and dissolved silica concentrations (mean values for samples collected in the photic zone of Lake Cumberland, Kentucky, in March-October 1994; 0-10 m). study. Ratios of N:P at Wolf Creek decreased from 8:1 in March to <1:1 during July-Octo- ber (atomic ratios based on NO, and TP de- terminations). Dissolved silica was also sub- stantially depleted in the upper water column as indicated by decreases in concentration from 3-4 mg/liter in March to 1-1.5 mg/liter by early July. Silica depletion was apparently greater at W-mid and W-up sites relative to downstream sites (W-dn and C-dn) as evi- denced by lower mid-summer silica concen- trations. Chlorophyll and nutrient data were ana- lyzed using a Generalized Linear Models Pro- cedure (SAS Version 6.03) to partition vari- ance arising from four sources: embayment (Caney and Wolf), sampling date (July, August, September), station (upstream, sidstretin, downstream), and depth (epilimnion, meta- limnion, and hypolimnion). In addition to the four main effects, the ANOVA model included three interaction terms: embayment with date (to determine if seasonal patterns differed be- tween embayments), embayment with station (to determine if longitudinal patterns differed between embayments), and embayment with depth (to determine if vertical profiles dif- fered between embayments). The empirical models were genet rally found to be good pre- dictors (p < 05) of variation in chlor ophyll (R2 = 0.52), pheophytin (R? = 0.55), nitrate (R? 0.75), and total phosphorus (R* = 0.48). The model describing variation in dissolved silica concentrations was not significant. Among the main effects, date and station were found to be significant predictors of chloro- phyll and pheophytin, date and depth were significant predictors of nitrate, and station was a significant predictor of total phosphorus (Table I). Embayment did not explain a sig- nificant proportion of the variance in mea- sured parameters; in only one model were any of the three interaction terms significant (em- bayment with depth in the chlorophyll model). DISCUSSION We interpreted our data for Lake Cumber- land in the context of the generalized reservoir 12 Transactions of the Kentucky Academy of Science 57(1) Results of statistical analyses partitioning variation in chlorophyll and nutrient concentrations associated with the effects of embayment, date, station, and depth (SAS Generalized Linear Model Procedure) in Lake Cumberland, Table 1. Kentucky. Source df Mean square Embayment 1 0.01 Date 2 15.21 Station 9} 14.16 Depth 2 11.00 Emb X Date 2 0.95 Emb X Station 2 1.10 Emb X Depth 2 18.25 Model 13 10.24 Error 31 3.99 Embayment 1 1,210 Date 2 26,532 Station 2 9,428 Depth 2 128,221 Emb X Date 2} 2,693 Emb X Station 2 388 Emb X Depth 2 977 Model 13 35,564 Error 29 5,231 Embayment 1 10.12 Date 2 6.79 Station 2 670.51 Depth 2 N59, Emb X Date 2 72.76 Emb X Station 2 26.91 Emb X Depth 2 60.01 Model 13 159.77 Error 29 TID model described by Kennedy and Walker (1990) and Kimmel, Lind, and Paulson (1990). Their model considers spatial heterogeneity in nutrient availability and phytoplankton pro- duction in relation to longitudinal gradients in water flow and basin morphology. A typical reservoir is considered to comprise three zones: uplake riverine zone, transition zone, and lacustrine (near dam) zone. The riverine zone is characterized by higher flow velocities, shorter water residence time, high concentra- tions of suspended solids, and low light pen- etration. The transition and lacustrine zones are characterized by increasing basin breadth and depth, decreasing flow, longer water res- idence time, and deeper light penetration (due to sedimentation of suspended solids). According to the model, nutrient concentra- tions are predicted to be highest in the river- ine zone due to elevated nutrient concentra- tions in watershed run-off and low rates of phytoplankton uptake under light-limiting conditions. Nutrient concentrations should de- F P Dependent variable 0.01 NS Chlorophyll 3.82 0.033 R? = 0.52 3.55 0.041 2.76 NS 0.24 NS 0.28 NS 4.58 0.018 2.57 0.015 0.23 NS Nitrate 5.07 0.013 R? = 0.75 1.80 NS 94.51 0.001 0.51 NS 0.07 NS 0.19 NS 6.80. 0.001 0.13 NS Total phosphorus 0.09 NS R? = 0.48 8.68 0.001 0.69 NS 0.94 NS 0.35 NS 0.78 NS 2.07 0.050 crease downstream (toward the dam) because greater light penetration allows for increased phytoplankton production. Spatial gradients in chlorophyll, total phos- phorus, and light penetration for Lake Cum- berland were consistent with predictions of the generalized model described above. Light penetration was lowest at upstream stations and increased with distance downstream. Overall, light availability did not appear to im- pose severe constraints on phytoplankton growth because the photic zone included the upper 8-12 m of the water column at all sites (maximum depth 15-25 m). Chlorophyll con- centrations were higher at upstream stations compared to samples collected downstream in the embayments and at the open-lake site. Light conditions and trends in chlorophyll suggest that our furthest upstream stations would be considered transition zone sites (area of high phytoplankton productivity) in the context of the generalized model. The Ca- ney Creek and Wolf Creek tributaries, rela- Water Quality in Lake Cumberland—Loy and Bukaveckas 13 tively small, were likely near base flow during most of the study period. It is therefore not surprising that we did not observe conditions typical of the riverine zone even at our fur- thest upstream sites. Spatial gradients in nutrient availability were more difficult to interpret because total phosphorus decreased downstream whereas nitrate and dissolved silica increased down- stream. The close correspondence between chlorophyll and total phosphorus concentra- tions suggests that phosphorus inputs via trib- utary streams may have stimulated phyto- plankton production in upstream areas. This would likely result in greater upstream deple- tion of nitrogen and dissolved silica and may account for the inverse gradients (increasing downstream) observed for NO, and SiO,. This argument assumes that phosphorus limits phy- toplankton production; however, our data sug- gest that the availability of nitrogen relative to phosphorus is very low in Lake Cumberland (N:P <10:1 at all sites during our study). Our measurements of phosphorus concentrations (based on TP) may overestimate phosphorus availability if the nonlabile (particle-bound) P fraction is large. Concentrations of suspended matter were low from May to September as indicated by low chlorophyll concentrations and low light attenuation coefficients. We feel, therefore, that our TP data are indicative of labile (dissolved) P in summer. Nitrogen avail- ability was inferred from measurements of ni- trate only; to the extent that NH, may con- tribute to the inorganic nitrogen pool in epi- limnetic waters, we may be underestimating N:P. For reservoirs where phytoplankton growth has been shown to be N-limited on the basis of physiological indicators (Groeger and Kimmel 1988), nitrogen availability was _re- ported to decrease downstream (opposite of the NO, gradients we observed in Lake Cum- berland). On the basis of data presented here, it is unclear which of these nutrients limits phytoplankton production in Lake Cumber- land. Bioassay experiments and additional measurements of P fractions (TP and SRP) would serve to clarify the relative importance of P and N in limiting phytoplankton produc- tion. We hypothesized that effects of recreational activities on water quality would be superim- posed on upstream-downstream gradients de- scribed above. Specifically, we predicted that nutrient and organic matter loading from houseboat discharge would result in elevated nitrogen, phosphorus, and chlorophyll concen- trations and greater hypolimnetic oxygen de- pletion in the Wolf Creek embayment. Con- sistently elevated TP levels measured at the station located immediately downstream (100 m) from a large marina complex (W-mid) sup- port this hypothesis. Although a corresponding increase in chlorophyll was not observed, higher rates of nitrate and silica depletion (in comparison to the downstream site) suggest greater phytoplankton production. Localized waste inputs containing inorganic or organic phosphorus could account for the observed trends in TP, NOs, and SiO,. An alternative hypothesis is that differences in TP between embayments are a result of differences in nu- trient inputs from associated tributaries. If tributaries were an important source of phos- phorus, we would expect that upstream sites would show a similar pattern of differences (Wolf Creek TP >Caney Creek TP). This was not the case as there were no consistent dif- ferences in TP concentrations at upstream sites. The midstream station in the Wolf Creek embayment occurs below the confluence of two smaller bays formed by the Wolf Creek and Alligator Creek tributaries. Water quality at this site may be influenced by inputs from Alligator Creek, which enter the reservoir 4.4 km (2.75 mi) upstream. As tributary sampling was not included in our study, we cannot ex- clude the possibility that elevated TP at this site was in part due to tributary influences. We compared dissolved oxygen profiles to test the hypothesis that organic matter loading resulted in greater hypolimnetic oxygen deple- tion in the Wolf Creek embayment. Oxygen saturation of 10 m samples collected at W-up and W-mid were consistently lower relative to corresponding sites in the Caney Creek em- bayment. Differences in oxygen resources be- tween the embayments are consistent with the hypothesis of higher organic matter loading in upper reaches of the Wolf Creek embayment. It is important to note, however, that stations W-up and W-mid were shallower (15 m) in comparison to corresponding sites at Caney Creek (25 m). Because the position of the thermocline was similar among sites (8-12 m), the volume of the hypolimnion in upstream 14 Transactions of the Kentucky Academy of Science 57(1) reaches of Wolf Creek was smaller in compar- ison to that of the Caney Creek sites. Thus, comparable rates of organic matter loading would result in greater apparent oxygen de- pletion per unit volume at shallower sites. In summary, our analyses of spatial variation in water quality revealed that the site near a major houseboat marina complex showed con- sistently elevated TP concentrations and high- er suspended particulate matter (as indicated by light scattering). Sites upstream and down- stream in the same embayment did not exhibit pronounced differences in water quality com- pared to sites in the other embayment, where houseboat activity was substantially lower. Overall, longitudinal gradients in both embay- ments followed expected patterns based on changes in flow velocity and basin morpholo- gy; our study does not provide compelling ev- idence of water quality deterioration resulting from houseboat discharge. This conclusion is supported by statistical models in which be- tween-embayment differences were not found to be significant predictors of variation in chlo- rophyll and nutrient concentrations. Our find- ings may be due in part to the low resolution of our sampling (monthly intervals), which is not suitable for detecting transient changes in water quality. Periods of intense houseboat ac- tivity, such as occur during the 4th of July hol- iday, may result in short-term reductions in water quality. These effects would not be de- tecting by our design and would require a more intensive field effort. ACKNOWLEDGEMENTS Support was provided by the University of Louisville’s Institute for the Environment and Sustainable Development in the form of an undergraduate research fellowship for D. Loy. We thank David Boger and Lake Cumberland State Dock for providing a boat and Rich Schultz for his analyses of nutrient samples. LITERATURE CITED [APHA] American Public Health Association. 1985. Stan- dard methods for the examination of water and waste- water. 16th ed. Washington, DC. Bukaveckas, P.A., and C.T. Driscoll. 1991. Effects of whole-lake base addition on the optical properties of three clearwater acidic lakes. Can. J. Fish. Aquatic Sci. 48:1030-1040. [DEP] Department of Environmental Protection. 1984. Trophic state and restoration assessments of Kentucky lakes, final report. Kentucky Natural Resources and En- vironmental Protection Cabinet, Frankfort. Edmondson, W.T. 1972. Nutrients and phytoplankton in Lake Washington. Pages 172-193 in G.E. Likens (ed). Nutrients and eutrophication. Am. Soc. Limnol. Ocean- ogr. Spec. Symp. 1. Groeger, A.W., and B.L. Kimmel. 1988. Photosynthetic carbon metabolism by phytoplankton in a nitrogen-lim- ited reservoir. Can. J. Fish. Aquatic Sci. 45:720-730. Kennedy, R.H., and W.W. Walker. 1990. Reservoir nutri- ent dynamics. Pages 109-132 in K.W. Thornton et al. (eds). Reservoir limnology: ecological perspectives. Wi- ley and Sons, New York. Kimmel, B.L., O.T. Lind, and L.J. Paulson. 1990. Res- ervoir primary production. Pages 133-194 in K.W. Thornton et al. (eds). Reservoir limnology: ecological perspectives. Wiley and Sons, New York. Kirk, J.T.O. 1983. Light and photosynthesis in aquatic ecosystems. Cambridge University Press, New York. Schindler, D.W. 1974. Eutrophication and recovery in ex- perimental lakes: implications for lake management. Science 184:897—899. Speziale, B.J., S.P. Schreiner, P.A. Giammatteo, and J.E. Schindler. 1984. Comparison of DMF, DMSO and ac- etone for extraction of phytoplankton chlorophyll. Can. J. Fish. Aquatic Sci. 41:1519-1522. Trans. Ky. Acad. Sei. 57(1):15-17. 1996. Survey of Cereal Aphids in Kentucky Wheat Fields: Common Species and Distribution Douglas W. Johnson Department of Entomology, University of Kentucky, Princeton, KY 42445-0469 and Donald E. Hershman Department of Plant Pathology, University of Kentucky, Princeton, KY 42445-0469 ABSTRACT Species determination and relative abundance of cereal aphids in samples from winter wheat were inves- tigated in major production areas of Kentucky in 1992-1993 and 1993-1994. Cereal aphids were more abundant in 1992-1993 than in 1993-1994. The bird cherry—oat aphid (Rhopalosiphum padi) was the most common aphid followed by English grain aphid (Sitobion avenae), corn leaf aphid (R. maidis), and greenbug aphid (Schizaphis graminum), respectively. All species were widely distributed in the Kentucky wheat pro- duction area. INTRODUCTION Barley yellow dwarf (BYD), a disease re- sulting from infection by a viral pathogen (BYDV), is the most important viral disease of cereals worldwide (Plumb 1983). The patho- gen is resident in a large number of crop and non-crop species of the grass family (Poaceae), e.g., corn (Zea mays) and tall fescue (Festuca arundinacea) and is vectored within and among these hosts/crops by a complex of aphid species (Homoptera: Aphididae). Fields of small grains infected with this disease often produce significantly reduced yields (Irwin and Thresh 1990). In Kentucky, BYD in wheat may be found at low levels each year; it oc- casionally reaches epidemic status. Much literature on BYD and associated hosts and vectors has been published by re- searchers around the world (Burnett 1989; Ir- win and Thresh 1990). In Kentucky, however, little information about BYD in production fields is available. We wish to establish which species of cereal aphids are most common in Kentucky soft red winter wheat (Triticum aes- tivum) fields and if the species vary from fall to spring and across the wheat-growing region. Specifically we wish to see if he situation in Kentucky is substantially similar to or different from that in surrounding areas. MATERIALS AND METHODS Aphid collections were made in fall and spring in each of the 1992-1993 and 1993-— 15 1994 growing seasons. Collection sites were selected to represent the distribution of wheat production in Kentucky (KAS 1991). Ten (1992-1993) and five (1993-1994) fields were sampled in each of 15 counties (Ballard, Bour- bon, Calloway, Christian, Daviess, Graves, Hardin, Henderson, Hickman, Logan, Shelby, Simpson, Todd, Trig ge, and Warren), producing a possible 150 and 75 samples, respectively (Figure 1). During both seasons fall samples were taken in the last 2 weeks of November and the first 2 weeks of December; spring samples were taken in April. The sample for each field was a composite of cereal aphids collected from foliage of five plants at five locations, randomly spaced along a diagonal line across the field, and were no dibker than ca. 15 m from the field edge or one another. Aphid populations were not over- ly abundant in either year, and no effort was made to differentiate the cereal aphids by lo- cation in a field. Aphids were obtained by col- lecting leaf sections on which they rested into appropriately labeled vials of 70% ETOH and held in the laboratory until examination. Iden- tification was accomplished by microscopic ex- amination and by reference to Pike, Boydston, and Allison (1991) and Stoetzel (1987). Sev- eral samples were discarded as unusable due to physical damage to the aphids from im- proper handling. 16 Transactions of the Kentucky Academy of Science 57(1) Figure 1. 1993-1994. RESULTS Cereal aphids were more abundant in 1992-1993 than in 1993-1994. Of the 150 fields visited in 1992-1993, the fall collection yielded 109 samples of which 104 were usable; of these, three contained mixed species. In spring, 109 samples were collected, and 100 were usable; of these, four contained mixed species. In 1993-1994, of the 75 fields visited, the fall collection produced only 17 samples yielding 16 usable, with one containing mixed species. The spring yielded 50 samples of which 49 were usable, with three containing mixed species. Four species of cereal aphids common in wheat production were found: Rhopalosiphum padi (L.), bird cherry—oat aphid; R. maidis (Fitch), corn leaf aphid; Schizaphis graminum (Rondani), greenbug aphid; and Sitobion ave- nae (Fab.), English grain aphid. All these spe- cies are able to serve as vectors for BYDV. Additionally, BYDV isolates associated with these aphids have been confirmed from Ken- Table 1. Numbers of samples containing various species of common cereal aphids, collected during a survey of soft red winter wheat fields in Kentucky in fall and spring 1992-1993 and 1993-1994. 1992-1993 1993-1994 Fall Spring Fall Spring Total samples 145 141 74 74 Samples with aphids 104 100 16 49 Bird cherry—oat aphid 99 85 16 3 Corn leaf aphid 1 0 0 0 English grain aphid 4 17 1 49 Greenbug aphid 3 0 0 0 Kentucky counties containing fields of soft red winter wheat sampled for cereal aphids in 1992-1993 and tucky soft red winter wheat fields (unpub- lished data). The bird cherry—oat aphid (BCOA) was the most commonly encountered species. It was collected in all four survey periods and was present in the greatest numbers of samples in three of the four collections. The English grain aphid (EGA) was the second most com- mon aphid. It was collected in all four collec- tion periods and dominated the samples in spring 1994. The corn leaf aphid (CLA) and greenbug aphid (GB) were found in much fewer numbers (Table 1). These cereal aphids are ubiquitous in Ken- tucky soft red winter wheat. When all samples are considered, BCOA was collected from ev- ery county surveyed; EGA was collected in all but four (Graves, Hardin, Hickman, and Simpson) of the 15 surveyed counties. The three GB samples were collected from Dav- iess and Graves counties; the two CLA sam- ples, from Ballard and Logan counties. DISCUSSION The species collected and their wide distri- bution in Kentucky soft red winter wheat were expected. Gildow (1987), in his summary of regional BYD research, pointed out that this same series of aphid species is commonly en- countered in Illinois, Indiana, Missouri, and many other states in the United States. How- ever, when working with a pathosystem as complex as BYD, unsubstantiated assumptions can quickly lead to erroneous results. For this reason and to allow for the use of research in Cereal Aphids in Kentucky Wheat Fields—Johnson and Hershman 17 nearby states it is important to know that these species are present. The commonness of BCOA is also to be ex- ected. However, the differences among BCOA, EGA, and CLA may be due, at least in part, to the timing of sample collection. CLA might be expected to occur in wheat ear- ly in fall and to decrease with the onset of cold weather; EGA is generally encountered late in spring after wheat begins to produce inflores- cences (personal observation). Our samples were generally collected later in fall and ear- lier in spring than one would expect to en- counter large numbers of CLA and EGA, re- spectively. A similar shift in species composi- tion was reported in Virginia (McPherson and Brann 1983). The large increase in EGA sam- ples in spring 1994 is probably due to the early occurrence of spring weather that year. Addi- tional research concerning the timing of shifts of aphid species in Kentucky wheat is certainly warranted. ACKNOWLEDGMENTS We thank Ms. Diane Perkins for collecting and maintaining the samples; the Kentucky Small Grain Growers Association for providing funds; the county extension agents for agri- culture for coordination of field selection and for obtaining permission to enter, and provid- ing directions to, survey fields in their coun- ties; and the several wheat producers for al- lowing us to work on their farms. The inves- tigation reported in this paper (95-08-066)is in connection with a project of the Kentucky Ag- ricultural Experiment Station and is published with the approval of the director. LITERATURE CITED Burnett, P.A. (ed). 1989. Barley yellow dwarf virus, the yellow plague of cereals. Centro Internacional de Me- joramiento de Maiz y Trigo, Mexico City. Gildow, F.E. 1987. Current status of barley yellow dwarf in the United States: a regional situation report. Pages 11-20 in P.A. Burnett (ed). Proceedings of the inter- national workshop: world perspectives on barley yellow dwarf, Udine, Italy, 6-11 July 1987. Irwin, M.E., and J.M. Thresh. 1990. Epidemiology of barley yellow dwarf: a study in ecological complexity. Annu. Rev. Phytopath. 28:393-424. [KAS] Kentucky agricultural statistics 1990-1991. 1991. Kentucky Agricultural Statistics Service, Louisville. McPherson, R.M., and D.E. Brann. 1983. Seasonal abun- dance and species composition of aphids (Homoptera: Aphididae )in Virginia small grains. J. Econ. Entomol. 76:272-274. Pike, K.S., L. Boydston, and D. Allison. 1991. Alate aphid vivipare associated with small grains in North America: a key and morphometric characterization. J. Kansas En- tomol. Soc. 63:559-602. Plumb, R.T. 1983. Barley yellow dwarf virus—a global problem. Pages 185-198 in R.T. Plumb and J.M. Thresh (eds). Plant virus epidemiology. Blackwell, London. Stoetzel, M.B. 1987. Information on the identification of Diuraphis noxia (Homoptera: Aphididae) and other aphid species colonizing leaves of wheat and barley in the United States. J. Econ. Entomol. 80:696—740. Trans. Ky. Acad. Sci. 57(1):18-21. 1996. Cryptosporidium parvum in the Domestic Dog Population of Central Kentucky Beverly W. Juett Department of Biological Sciences, Midway College, Midway, Kentucky 40347 Raymond B. Otero Department of Biological Sciences, Eastern Kentucky University, Richmond, Kentucky 40475 and William H. Bischoff Fayette Veterinary Clinic, 144 East Reynolds Road, Lexington, Kentucky 40503 ABSTRACT Cryptosporidium parvum oocytes were recovered from fecal specimens of 17 of 100 healthy or diseased adult and puppy dogs tested in central Kentucky. The specimens were collected and placed in 10% formalin, concentrated by the formalin-ethyl acetate method, and stained by a modified hot acid-fast technique. The presence of oocysts in domestic dogs may represent a possible reservoir that could infect immunocomprom- ised and/or immunocompetent humans. INTRODUCTION Cryptosporidium parvum is an enteric coc- cidian protozoan assigned to phylum Apicom- plexa, class Sporozoasida, and order Eucocci- diorida (Current 1985). In the suborder Emeriorina, C. parvum is found with three other important human pathogens, Toxoplas- ma gondii, Isospora Sie, and Sarcocystis spp. (Current 1985; Dubey, Speer, and Fayer 1990). The life cycle of C. parvum exhibits asexual and sexual endogenous stages; oocysts are discharged in the feces of the Hest (Dubey, Speer, and Fayer 1990; Fayer and Ungar 1986; Loose, Sedergran, and Cooper 1989; Tzipori 1988). Oocysts are found in the general environ- ment including water and food (Hayes et al. 1989; Tzipori 1988). Widespread in nature, C. parvum infects many animals (Fayer and Un- gar 1986). Spread by the oral-fecal route, it has been implicated as a zoonosis (D’ Antonio, Winn, and Zajac 1985; Hayes et al. 1989). Cryptosporidiosis, reported to cause disease in both immunocompetent (CDC 1982; Cur- rent et al. 1983; F ayer and Ungar 1986; Soave 1988; Tzipori 1987, 1988) and immunocom- promised humans (CDC 1982; Current et al. 1983; Soave and Johnson 1988; Tzipori 1987), is a significant source of morbidity in both groups. In the immunocompromised patient it 18 is a source of severe prolonged diarrhea. No specific effective treatment is available at the present time (Cotton 1991: Soave and Arm- strong 1986), although paromycin has been used with some success (Armitage et al. 1992; Bissuel et al. 1994; Cotton 1991; Goodgame et al. 1993). Therapy studies are difficult to design because patients are severely immu- nosuppressed and may have multiple intestinal infections. Rat models indicate successful treatment with paromycin at higher dosages than the 1-2 g/day human patients usually re- ceive (Verdon et al. 1995). Associations between calves and human dis- ease are firmly established (Rahaman et al. 1984). Evidence suggests a link between do- mestic cats and cryptosporidiosis in humans (Fayer and Ungar 1986). Puppies have been suspected as a source of Cryptosporidium oo- cysts (Current et al. 1983). Serological studies suggest that cryptosporidiosis in dogs is a fre- quent occurrence (Tzipori and Campbell 1981). Several surveys have been conducted on domestic dogs to detect C. parvum oocysts (Chermette and Blondell 1989; Fayer and Un- gar 1986; Pohjola 1984). Dogs excreting the oocysts may be asymptomatic carriers or may exhibit disease symptoms such as diarrhea. Because of the lack of treatment for cryptos- poridiosis, and because of the severe discom- Cryptosporidium parvum—Juett, Otero, and Bischoff 19 fort seen in immunocompromised patients, guidelines for these patients in regard to their association with pet dogs may ee to be es- tablished. Similar guidelines concerning food counseling for persons infected with HIV have been issued by the U.S. Public Health Service (Archer 1989). These state that the risk of con- tracting cryptosporidiosis for HIV patients can be reduced by avoiding raw foods of animal origin and by washing all foods to avoid con- tamination from soil. MATERIALS AND METHODS Specimens Fecal specimens were collected from 100 domestic, diseased or healthy adult and puppy dogs in central Kentucky in September, Oc- tober, and early Nov ember 1991 and 1992. Af- ter the specimens were collected or had ar- rived at the veterinary clinic, they were placed in a preservative (10% formalin) in screw- cap- ped containers. A questionnaire, designed to gather pertinent information, asked the dog's name, breed, age, sex, illnesses at the time, and whether the dog had an active case of di- arrhea. Of the 100 dogs, 40 were males, 56 were females, and 4 were of unknown sex: 49 were | year of age or less, 33 were 1-10 years, 11 were over 10 years, and 8 were of unknown age; and 81 were well dogs and 19 had re- ported illnesses. Processing Methods The stool specimens were concentrated us- ing a modified formalin-ethyl acetate proce- dure (Baron, Peterson, and Finegold 1994) based on Ritchie’s s methdology (Ritchie 1948). The concentrate was spread thinly on a pre- cleaned slide and heat fixed at 70°C for 10 minutes before staining. The smears were stained with a modified Rot Ziehl-Neelsen car- bol fuchsin (ZN) for 5 minutes while heating the smear to steaming. The decolorizer was 5% H,SO, for 30 Seconds the counterstain, methylene blue for 1 minute. Stained smears were evaluated by bright field microscopy at 1000 for Cryptosporidium oocysts. Oocysts stain bright red. Yeasts have been reported to stain blue by some researchers, but many of the yeasts ‘encountered in our specimens stained ted. Therefore, the size of the object was important in distinguishing oo- cysts from yeasts. Cryptosporidium parvum is 4—6 jum; many yeasts are smaller. To distinguish the yeast that was in the size range for C. parvum, an iodine stain was used on “all specimens where the ZN smears ap- peared red. When exposed to iodine for less than 15 minutes, yeasts stain but C. parvum does not (Baron, Peterson, and Finegold 1994: Ma and Soave 1983). Therefore, the criteria to call a specimen positive for C. parvum oo- cysts included (1) bright red color on modified hot ZN with the object viewed falling into the 4—6 um range and (2) concentrated sediment showing stajenures comparable in size that did not stain with iodine. Each batch of acid-fast stains included a positive control for C. par- vum purchased from Trend Scientific, Inc., St. Paul, Minnesota 55112. Each control was fod with the specimens; satisfactory results were demonstrated for each control. RESULTS Cryptosporidium parvum oocysts were present in 17 fecal specimens. Of the seven dogs with oocysts nine were female, seven were male, and one was in the unknown cat- egory (fecal specimen delivered to clinic with- out information). These dogs represent 16% of the female population and 18% of the male population, The infected dogs ranged from a few weeks to 12 years old. Fifteen were 1 year or under, one was 3 years, and one was 12 years. Of the 100 dogs examined, 31% of age 1 year and under were positive for oocysts, 3% of age 2 to 10 years were positive, and 9% of age 10 years s and over were positive. Oocysts were found in two dogs with diar- rhea, but nine dogs with diarrheal symptoms were negative. F ecal specimens from 81 ap- parently healthy dogs revealed 15 cases of cryptosporidiosis. It was difficult to tabulate information concerning the breeds of dogs represented. In the entire population, 41 breeds were represented by at least one ani- mal. Of these, only English setters and Ger- man shepherds showed more than one posi- tive case. Two out of three English setters (66%) and four of four (100%) German shep- herds demonstrated oocysts. DISCUSSION We think that ours is the first report of C. parvum oocysts from dogs in central Kentucky 20 Transactions of the Kentucky Academy of Science 57(1) (where the incidence was 17%) and the first comprehensive analysis of a cross section of domestic dogs in the United States. Dogs de- velop natural infections with C. parvum (Cur- rent et al. 1983), but the prevalence rate for dogs is largely unknown (Dubey, Speer, and Fayer 1990). The group of dogs demonstrating the high- est incidence was | year old and under. In the study by Current et al. (1983), puppies were suspected as a source of infection for crypto- sporidiosis; the high incidence rate in young dogs from central Kentucky appears to sup- port this speculation. The relatively high rate of C. parvum cases in the domestic dog population in our study suggests a possible reservoir for oocysts that might infect immunocompromised and im- munocompetent humans. Because most dogs exhibiting C. parvum oocysts were asympto- matic, their owners would probably consider them healthy and not a potential source of in- fection. With only two dogs (12%) testing pos- itive and showing diarrheal symptoms, such symptoms cannot be used to implicate a dog as a reservoir. Geographical factors may have an effect on positive rates. We had insufficient data to de- termine the proximity of positive cases to each other. Clustering effects may have occurred with dogs from the same household. In future studies, this factor would need to be delineat- ed more clearly. Since this is thought to be the first compre- hensive study on a cross section of domestic dogs in central Kentucky, it remains to be de- termined if our data are representative of dog populations in general. More research on pop- ulations in Kentucky and other states is need- ed before definitive prevalence rates can be determined conclusively. Cryptosporidiosis is a significant threat to immunocompromised individuals such as AIDS patients. Public health statistics released 30 Jun 1995 in the Kentucky AIDS Surveil- lance Report (HSD 1995) listed 189 cases (11.5% of total AIDS cases) of HIV Wasting Syndrome from 1982 with 11.5% of all Ken- tucky AIDS patients exhibiting this as primary disease. Cryptosporidiosis remains largely un- treatable except for limited success with pa- romycin. Therefore, prevention may be the best path to follow for immunocompromised individuals. Public health guidelines for pre- venting listeriosis and salmonellosis in such patients (Archer 1989) include avoiding raw milk and eggs and washing fruits and vegeta- bles before eating. If prevalence rates were better delineated for domestic dogs and the results of our study are supported in the fu- ture, guidelines may include avoidance of dogs for these patients as one method for prevent- ing cryptosporidiosis. ACKNOWLEDGEMENTS We thank D. M. Mardon and J. P. Harley of Eastern Kentucky University for sugges- tions and the staff at the Fayette Veterinary Clinic on Reynolds Road in Lexington, Ken- tucky, for help in collecting specimens. This work was supported by the departments of bi- ology at Eastern Kentucky University and Midway College. LITERATURE CITED Archer, D.L. 1989. Food counseling for persons infected with HIV. Public Health Rep. 104:196—198. Armitage, K., T. Flanigan, J. Carey, I. Frank, RR. MacGregor, P. Ross, R. Goodgame, and J. Tumer. 1992. Treatment of cryptosporidiosis with paromycin. Arch. Internal Med. 152:2497—2499. Baron, E., L. Peterson, and S. Finegold. 1994. Bailey and Scott’s diagnostic microbiology. 9th ed. C.V. Mosby, St. Louis. Bissuel, F., L. Cotte, M. Rabodoniria, P. Rougier, M. Piens, and C. Trepo. 1994. Paromycin: an effective treatment for cryptosporidial diarrhea in patients with AIDS. Clin. Infect. Dis. 18:447-449. [CDC] Center for Disease Control. 1982. Cryptospori- diosis: assessment of chemotherapy of males with ac- quired immune deficiency syndrome (AIDS). Morbid. Mortal. Weekly Rep. 31:589-592. Chermette, R., and S. Blondell. 1989. Cryptosporidiose des carnivores domestiques: résultats préliminaires en France. Bull. Soc. Frang. Parasitol. 7:31—36. Cotton, P. 1991. Medicine’s arsenal in battling ‘dominant dozen’, other AIDS-associated opportunistic infections. J. Am. Med. Assoc. 266:1476-1481. Current, W.L. 1985. Cryptosporidiosis. J. Am. Vet. Med. Assoc. 187:1334-1338. Current, W.L., N.C. Reese, J.V. Ernst, W.S. Bailey, M.B. Heyman, and W.M. Weinstein. 1983. Human crypto- sporidiosis in immunocompetent and immunocom- promised persons. New Engl. J. Med. 308:1252—1257. D’Antonio, R., R. Winn, and R. Zajac. 1985. Sequential acute gastroenteritis from contaminated drinking water caused by Norwalk virus and Cryptosporidium. Clin. Res. 33:399A. Dubey, J., C. Speer, and R. Fayer (eds). 1990. Cryptospo- Cryptosporidium parvum—Juett, Otero, and Bischoff 21 ridiosis of man and animals. CRC Press, Boca Raton, Florida. Fayer, R., and B.L.P. Ungar. 1986. Cryptosporidium spp. and cryptosporidiosis. Microbiol. Rev. 50:458-483. Goodgame, R., R. Genta, A.C. White, and C. Chappell. 1993. Intensity of infection in AIDS-associated cryp- tosporidiosis. J. Infect. Dis. 167:704—709. Hayes, E.B., T.D. Matte, T.R. O’Brien, T.W. McKinley, (Cask Logsdon, J.B. Rose, B.L.P. Ungar, D.M. Word, PF. Pinsku, M.L. Cummings, M.A. Wilson, E.G. Long, E.S. Hurwitz, and D. Juranek. 1989. Large community out- break of cryptosporidiosis due to contamination of a fil- tered public water supply. New Engl. J. Med. 320: 1372-1376. [HSD] Health Services Department, Division of Epide- miology, State of Kentucky. 1995. Kentucky AIDS Sur- veillance Report. June 30. Frankfort. Loose, J.H., D.J. Sedergran, and H.S. Cooper. 1989. Identification of Cryptosporidium in paraffin-embed- ded tissue sections with the use of a monoclonal anti- body. Am. J. Clin. Pathol. 91:206—209. Ma, P., and R. Soave. 1983. Three-step stool examination for cryptosporidiosis in 10 homosexual men with pro- tracted watery diarrhea. J. Infect. Dis. 147:824—828. Pohjola, S. 1984. Survey of cryptosporidiosis in feces of normal healthy dogs. Nordic Vet. Med. 36:189—-190. Rahaman, A., S.C. Sanyal, K.A. Al-Mahmyd, A. Sobhan, K.S. Hossain, and B.C. Anderson. 1984. Cryptospori- diosis in calves and their handlers in Bangladesh. Lan- cet 2:22]. Ritchie, L.S. 1948. An ether sedimentation technique for routine stool examinations. Bull. U.S. Army Med. Dep. 8:326. Soave, R. 1988. Cryptosporidiosis and isosporiasis in pa- tients with AIDS. Infect. Dis. Clin. North Am. 2:485— 493. Soave, R., and W.D. Johnson. 1988. Cryptosporidium and Isospora belli infections. J. Infect. Dis. 157:225— 229. Soave, R., and D. Armstrong. 1986. Cryptosporidium and cryptosporidiosis. Rev. Infect. Dis. 8:1012—1023. Tzipori. S. 1987. Annotation: cryptosporidiosis in child- hood. Austral. Paediat. J. 23:89-91. Tzipori, S. 1988. Cryptosporidiosis in perspective. Adv. Parasitol. 27:63-129. Tzipori, S., and I. Campbell. 1981. Prevalence of Cryp- tosporidium antibodies in 10 animal species. J. Clin. Mi- crobiol. 14:455—456. Verdon, R., J. Polianski, C. Gaudebout, and J. Pocidalo. 1995. Paromycin for cryptosporidiosis in AIDS. J. In- fect. Dis. 171:1070. Trans. Ky. Acad. Sci. 57(1):22-26. 1996. Age, Growth, and Food of Freshwater Drum, Aplodinotus grunniens (Sciaenidae), in Kentucky Lake, Kentucky/Tennessee David P. Dreves, Tom J. Timmons, and Jeffrey Henson Hancock Biological Station, Murray State University, Murray, KY 42071 ABSTRACT Zebra mussels (Dreissena polymorpha) have recently been discovered in the lower Tennessee River. In- formation on age and growth rates of freshwater drum in Kentucky Lake, Kentucky/Tennessee, before colonization of zebra mussels is important, especially if freshwater drum were to be managed to control expansion of this rapidly spreading exotic. Age and growth of freshwater drum in Kentucky Lake were determined from fish supplied by local commercial fishermen in 1985 and 1986. Growth in the reservoir was generally slower than in other bodies of water. The diet of freshwater drum was also examined to determine feeding habits. The fish were feeding on the exotic mussel Corbicula fluminea. Zebra mussels in the diet of freshwater drum have already been documented for Lake Erie. These facts suggest that once zebra mussels colonize Kentucky Lake, freshwater drum will be a substantial predator on them. INTRODUCTION The freshwater drum (Aplodinotus grun- niens, Sciaenidae) is a common fish of streams, rivers, and reservoirs in much of midwestern North America. Because of its strong molari- form pharyngeal teeth, the fish is capable of feeding on mollusks (Moen 1955; Price 1963; Wrenn 1968). With the recent invasion of the zebra mussel (Dreissena polymorpha) into North America, the freshwater drum may play an important role as a biological control of this rapidly spreading exotic. French and Bur (1993) showed that freshwater drum greater than 250 mm in length fed on zebra mussels in Lake Erie. They suggested that anglers should release all freshwater drum to increase biological control of the mussels. French (1993) recommended that fishery managers in eastern North America should manage for na- tive molluscivores to suppress zebra mussel populations. He recommended research on prey-preference and mollusk-eating efficiency by freshwater drum because their pharyngeal teeth make them the most efficient zebra mussel predator. Zebra mussels were first reported in the lower Ohio and Tennessee rivers in autumn 1991 (Sickel and Leek 1994). Since that time they have been found on native mussels in Kentucky Lake, Kentucky/Tennessee, in the area of Camden, Tennessee. In July 1992 a cluster of seven adult zebra mussels was found near the west shore of Kentucky Lake at Ten- 22 nessee River Mile (TRM) 23.8. In autumn 1992 the locks at Kentucky Dam on the Ten- nessee River were dewatered for mainte- nance. The locks contained many attached ze- bra mussels, which probably came from barges and other vessels traveling from areas where the mussel populations are denser (Sickel and Leek 1994). By autumn 1993 the lower Ohio River had been completely colonized by zebra mussels; the lower Tennessee River remained relatively uncolonized. As freshwater drum may become important to Kentucky Lake fishery managers, informa- tion on age and growth rates of this species before colonization by zebra mussels would be valuable if freshwater drum were to be man- aged to control expansion of zebra mussel populations. Also, Kentucky Lake presently supports a substantial commercial fishery for buffalo (Ictiobus spp.), common carp (Cypri- nus carpio), and catfish (Ictalurus spp.). Freshwater drum is the most common non- target species in the commercial catch in Ken- tucky Lake, but currently no market exists (Timmons et al. 1989). If a commercial fishery for this species was ever developed, growth data would be important for determining gear and harvest regulations. Several authors studied freshwater drum in Tennessee River impoundments (Dendy 1946; Wrenn 1968), but we are not aware of any study of Kentucky Lake populations. The ob- jectives of our research were to describe the Freshwater Drum in Kentucky diet of freshwater drum and to examine the age and growth of this fish in Kentucky Lake from samples collected prior to colonization by zebra mussels. These data may be \ valuable in assessing the effect of zebra ‘mussel colo- nization on the freshwater drum population in Kentucky Lake. MATERIALS AND METHODS Kentucky Lake is in western Kentucky and western Tennessee on the Tennessee River. This reservoir was formed by the impound- ment of the Tennessee River at Tennessee River mile (TRM) 22 the reservoir is ca. 64,800 ha with 21,000 ha within Kentucky and 43,800 ha within Ten- nessee. Freshwater drum were collected from throughout the reservoir, stretching from Ken- tucky Dam (TRM 22) to Pickwick Dam (TRM 207). Freshwater drum used for age determina- tion were collected in 1985 and 1986 by com- mercial fishermen using gill nets, baitlines, snaglines, and hoopnets in Kentucky Lake. Small individuals not present in the commer- cial harvest were collected concurrently by us- ing small mesh gill nets (<76 mm). Total length (TL) and weight were recorded _ for each fish. Scales were removed from the left side of each fish for age determination, placed on a heated acetate slide, and pressed with a Wildco laboratory press. Scale impressions were viewed using a Bell and Howell Model ABR-IV microfiche reader and measured from the focus to each annulus and the scale margin along the anterior-median primary radius. An- nuli were determined by using the standard criteria of crowding and crossing over of cir- culi (Lagler 1956). Lengths at previous annuli were calculated using the Fraser-Lee method (Bagenal and Tesch 1978). The correction fac- tor “a” was extrapolated with a length-margin linear regression model and was equal to the Y-intercept. . The fish used for diet analysis were collect- ed in a 12-month period beginning in Novem- ber 1987 and ending in November 1988. The stomachs of 122 freshwater drum were re- moved and examined. Food items were iden- tified and the percent frequency of occurrence for several freshwater drum size classes was calculated by dividing the number of stomachs Lake in 1944. Surface area of Dreves, Timmons, and Henson 93 containing the item by the number of non- empty stomachs. RESULTS Scales from 322 freshwater drum were aged and measured for backcalculation. Age classes represented in the sample ranged from 0 to 13 with one fish at 16. Fish lengths at capture ranged from 86 to 775 mm. Linear regression of LE on mae resulted in the equation length = 64.49 + 2.27 margin, which was sig- a Geaut at p<0.0001, r? = 0.914. The Y-inter- cept (64.49) was used as the correction factor in the Fraser-Lee formula for backcalculation of length at annulus formation. The first year of growth of freshwater drum in Kentucky Lake was rapid, with a mean length at age 1 of 130 mm (Table 1). Growth rate was consistent between ages | and 10, averaging 41 mm/year. Freshwater drum av- eraged 503 mm at age 10 and grew at an av- erage rate of 31 mm/year from age 10 through 13. The length-weight equation log,, weight = —6.10 + 3.46 log, length was significant at p<0. 0001, r?2 = 0.951. Kentucky Lake freshwater drum fed _pri- marily on benthic or ganisms (Table 2). Small fish from 100 to 300 mm fed about equally on larvae of Ephemeroptera (Hexagenia) and Diptera. The Asiatic clam Corbic ita fluminea appeared in stomachs of only 2.0% of the small fish. However, after the fish attain 300 mm, the diet shifted from insect larvae to Cor- bicula, with the shift becoming more pro- nounced with increasing fish length. The fre- quency of occurrence of Asiatic clams was 71% for individuals greater than 300 mm and 84% for those greater than 400 mm. Corbicula was often the only food of fish longer than 500 mm. Crayfish were occasionally consumed by larger freshwater drum: fishes as prey were present in low numbers in all freshwater drum size classes. DISCUSSION The growth of freshwater drum in Kentucky Lake was generally slower than that reported in many other waters (Table 3). The differ- ences in growth rates do not appear to be re- lated to climate as some of the populations exhibiting more rapid growth than in Ken- tucky Lake are more southern in latitude (Benton, Jackson, and Davies 1988; Houser 24 Table 1. collected from Kentucky Lake in 1985-1986. Transactions of the Kentucky Academy of Science 57(1) Mean backcalculated lengths (mm) and weighted mean increment of growth (mm) for 322 freshwater drum Mean length at annulus formation Age n 1 2 3 4 5 6 th 8 9 10 11 12 13 14 15 16 0) 10 ] 28 117 2 52 125 161 3 46 125 169 201 4 60 128 172 206 241 5 41 131 176 212 244 279 6 16 131 178 216 248 284 322 ih 28 139 186 219 257 293 328 364 8 18 131 176 214 251 291 331 365 4i11 9 7 141 187 221 259 288 333 367 402 443 10 6 142 180 221 279 323 363 405 451 510 561 11 6 138 189 231 272 307 355 395 431 488 539 578 12 2 138 191 250 284 327 352 376 412 452 498 566 595 13 1 115 151 171 210 261 285 349 391 431 468 499 510 535 14 Oc — —S =—_ SS 15 0 — 16 1 121 155 192 228 253 289 337 396 416 448 477 577 657 677 697 722 Weighted mean 130 175 213 252 291 329 370 413 457 503 530 561 596 677 697 722 Weighted mean increment Abra Soe eo) o> se Za CRP ee iy i Bh a) IL DS 1960) and one is more northern (Butler and Smith 1950). Freshwater drum in several nat- ural northern lakes have faster growth rates early in life and slower growth rates later in life compared to those in Kentucky Lake (Bur 1984; Priegel 1969). The growth of freshwater drum in Kentucky Lake most closely resem- bled that of this species in two other reser- voirs. Wheeler Reservoir (Wrenn 1968), an- other Tennessee River impoundment is more southern, located in Alabama. Kentucky Lake fish grew faster initially, but by 5 years of age, fish from Wheeler Reservoir had surpassed the growth of freshwater drum in Kentucky Table 2. Percent frequency of occurrence of food items from freshwater drum collected from Kentucky Lake in 1987-1988. n = 122. (sample size in parentheses). Total length (mm) 100-199 200-299 300-399 400-499 =500 Food (23) (52) (16) (12) (19) Corbicula sp. — 20 625 83.3 84.2 Crayfish — — 195 <= 105 Diptera (larvae) EO 0.0) he Ephemeroptera (larvae) 47.8 49.0 37.5 16.7 — Odonata (larvae) — BE) s= 83a Miscellaneous insects — — — 33a Miscellaneous fishes 43 78 125 83 10.5 Lake and continued to widen the differential with increasing age. Lewis and Clark Lake (Swedberg 1965) is a more northern reservoir, located on the Nebraska—South Dakota bor- der. Growth rates there were similar to the present study, although fish in Kentucky Lake lived slightly longer. Freshwater drum feed on clams and mus- sels after reaching some size threshold be- tween 250 and 300 mm TL. In Kentucky Lake, fish greater than 300 mm TL fed exten- sively on the Asiatic clam, often the only food of fish longer than 500 mm TL. Native mus- sels are also occasionally consumed by fresh- water drum in the lake (Paul Rister, pers. comm.). Individuals between 100 and 300 mm TL fed on Hexagenia and dipteran larvae. Food habits reported here (Table 2) are sim- ilar to those reported from Wheeler Reservoir (Wrenn 1968). In both reservoirs, freshwater drum less than 254 mm TL fed predominantly on dipteran larvae, with copepods and the lar- vae of Ephemeroptera and Plecoptera of less- er importance. Freshwater drum greater than 254 mm TL fed predominantly on Corbicula and fishes in Wheeler Reservoir (Wrenn 1968). The same pattern was evident more re- cently in Lake Erie with freshwater drum Freshwater Drum in Kentucky Lake—Dreves, Timmons, and Henson 25 Table 3. Tennessee 1985-1986, and other studies. l D 3 Kentucky Lake (present study) 130m eel lS Jordan Dam Tailrace, AL Benton, Jackson, and Davies (1988)! 148 214 269 U. Mississippi River Butler & Smith (1950) WL Aa}8) —BXS\7/ Lake Erie Bur (1984) males Viel SOme225 females 190 227 Oklahoma Reservoirs Houser (1960)? 107 216 300 Wheeler Reservoir, AL Wrenn (1968) 81 145 198 Lake Winnebago, WI Priegel (1969) ISO 1S aris Lewis and Clark Lake, NE-SD Swedberg (1965) 94 166 209 ' Unweighted mean. Mean backcalculated lengths (mm) at each annulus for freshwater drum in Kentucky Lake, Kentucky and Length at each annulus 4 5 6 i 8 9 10 1] 12 252 291 329 370 413 457 503 530 561 312 362 406 444 456 479 340 376 419 460 485 251 278 307 334 359 407 451 209 287 315 351 382 425 488 567 353 389 467 518 572 592 635 678 754 241 295 348 406 475 551 607 647 676 312 338 356 368 381 391 427 2Al 271 311 355 410 451 502) 9573 2 Unweighted mean from four reservoirs with the sample size from each greater than 100 fish feeding on zebra mussels. French and Bur (1993) found that predation on zebra mussels occurs in fish greater than 250 mm and in- creases with total length. Zebra mussels were the most abundant food item by volume and dry weight in both medium (250-374 mm TL) and large (375-574 mm TL) fish, with some larger specimens eating almost exclusively ze- bra mussels. They believe freshwater drum will likely feed on zebra mussels wherever their ranges coincide. If zebra mussels reach densities in Ken- tucky Lake similar to those reported for Lake Erie, the freshwater drum population in Ken- tucky Lake may be profoundly affected. We predict that freshwater drum will be a signif- icant predator of zebra mussels when the mus- sels colonize Kentucky Lake. This may cause an increase in the numbers, growth rate, and production of freshwater drum, already the most common incidental fish in the Kentucky Lake commercial catch (Timmons et al. 1989). Whether they will select thin shelled zebra mussels over thicker shelled Corbicula is not known. We are not aware of any studies doc- umenting the effects of freshwater drum pre- dation on zebra mussel populations, an area certainly in need of further study. LITERATURE CITED Bagenal, T.B., and F.W. Tesch. 1978. Age and growth. Pages 101-136 in T.B. Bagenal (ed). Methods for as- sessment of fish production in fresh waters. 3rd_ ed. Blackwell Scientific Publications, Oxford, Great Britain. Benton, J.W., D.C. Jackson, and W.D. Davies. 1988. Characteristics of the freshwater drum fishery below Jordan Dam. Proc. Annu. Conf. Southeast. Assoc. Fish Wildl. Agencies 42:218—224. Bur, M.T. 1984. Growth, reproduction, mortality, distri- bution, and biomass of freshwater drum in Lake Erie. J. Great Lakes Res. 10:48-58. Butler, R.L., and L.L. Smith Jr. 1950. The age and rate of growth of the sheepshead, Aplodinotus grunniens Rafinesque, in the Upper Mississippi River navigation pools. Trans. Am. Fish. Soc. 79:43—54. Dendy, J.S. 1946. Food of several species of fish, Norris Reservoir, Tennessee. J. Tennessee Acad. Sci. 21:105— WE French, J.R.P., HI. 1993. How well can fishes prey on zebra mussels in eastern North America? Fisheries 18: 13-19. French, J.R.P., If, and M.T. Bur. 1993. Predation of the zebra mussel (Dreissena polymorpha) by freshwater drum in western Lake Erie. Pages 453-464 in T.F. Na- lepa and D.W. Schloesser (eds). Zebra mussels: biology, impacts, and controls. Lewis Publishers, Boca Raton, FL. Houser, A. 1960. Growth of freshwater drum in Okla- homa. Oklahoma Fish. Res. Lab. Rep. 78. 26 Transactions of the Kentucky Academy of Science 57(1) Lagler, K.F. 1956. Freshwater fishery biology. 2nd ed. William C. Brown Company, Dubuque, IA. Moen, T. 1955. Food of the freshwater drum, Aplodi- notus grunniens Rafinesque, in four Dickinson County, Iowa, lakes. Proc. Iowa Acad. Sci. 62:589-598. Price, J.W. 1963. A study of the food habits of some Lake Erie fish. Ohio Biol. Surv. Bull. 2. Priegel, G.R. 1969. Age and rate of growth of the fresh- water drum in Lake Winnebago, Wisconsin. Trans. Am. Fish. Soc. 98:116-118. Sickel, J.B., and D.A. Leek. 1994. Observations on zebra mussel colonization in the lower Ohio and Tennessee rivers. Pages 447-455 in A.H. Miller (ed). Proceedings 4th International Zebra Mussel Conference 94, Wis- consin Sea Grant Institute, Madison, WI. Swedberg, D.V. 1965. Age and rate of growth of fresh- water drum, Lewis and Clark Lake, Missouri. River. Proc. South Dakota Acad. Sci. 44:160-168. Timmons, T.J., R.S. Hale, T.L. Hoffnagle, and J.B. Soldo. 1989. Incidence of sport fishes in the commercial fish catch from Kentucky Lake, Kentucky and Tennessee. North Am. J. Fish. Manage. 9:209-212. Wrenn, W.B. 1968. Life history aspects of smallmouth buffalo and freshwater drum in Wheeler Reservoir, Al- abama. Proc. Annu. Conf. Southeast. Assoc. Game Fish Comm. 22:479-495, Trans. Ky. Acad. Sci. 57(1):27-32. 1996. Understanding the Mass Distribution of the Dwarf Galaxy DDO 170 M. K. Falbo-Kenkel and K. L. Kremer Department of Physics and Geology, Northern Kentucky University Highland Heights, KY 41099 ABSTRACT Using a two-component mass model consisting of (1) and ( a luminous disk with constant mass to light ratio 2) an HI gas disk, we determined the rotation curve for the dwarf galaxy DDO 170 in the conformal model of gravity. The rotation curve we derived using the two-component, “maximal disk” mass model is in excellent agreement with the measured rotation curve. INTRODUCTION Compared to studies of mass distribution in bright spiral galaxies, much less is known about the distribution of mass in dwarf spiral and irregular galaxies. Recent studies of dwarf galaxies have added to the evidence that dark matter is a significant contributor to the mass of these galaxies (Broeils 1992; Carignan and Beaulieu 1989; Lake 1989: Lake, Schommer, and van Gorkom 1990). Mass models for most dwarf galaxies studied to date include three components (1) a luminous disk with a con- stant mass to light ratio, (2) a gas disk, and (3) a dark matter halo. As is the case for bright spiral galaxies, dark matter distributions for dwarf spiral and irreg- ular galaxies can be determined by comparing the “measured mass” of stars and gas with the dynamical mass via a comparison of the rota- tion curve predicted using the measured mass, and of the rotation curve determined by mak- ing measurements of the Doppler shifts of light from the galaxy at several radii. The well- documented result of this mass comparison in- dicates the existence of dark matter (Binney and Tremaine 1987). The specific distribution of dark matter in every galaxy is different, but in general the dark matter makes up 50% to 90% of bright galaxies and is distributed in an isothermal sphere or halo. The basic halo pa- rameters (halo core aan. phase density, and asymptotic velocity) are very sensitive to the mass model used to describe the galaxy. Even though observation of a galaxy and measure- ments of its luminosity aa total flux indicate the general distribution of stars and gas, re- spectively, the overall mass of the galaxy can- not be determined with great precision, and bo ~l the ratios M,,/M,.,, and M,,../L are variable parameters in most rotation curve fits. These factors lead to the somewhat unsettling con- clusion that the percentage and distribution of dark matter and the ratio M,,/M,,,. vary widely from galaxy to galaxy for no fundamental phys- ical reason. Based upon the many unanswered ques- tions concerning the nature of dark matter and the fact that there seems to be a great deal of arbitrariness when determining the amount and distribution of dark matter in galaxies, it is reasonable to ask whether or not two-com- ponents mass models can be used to under- stand the mass distributions in galaxies and, if so, how. These are the questions in which we are interested. We chose to study DDO 170 for several reasons. The most important of these was the high quality of the HI images and photometry that has led to a well-determined rotation curve for this particular dwarf galaxy. In ad- dition, to study the mass distribution of a gal- axy using galactic rotation as the comparator, we note that the galaxy must be relatively iso- lated so that the perturbative effects of neighbors are minimal. The gas distribution in the galaxy must be smooth and symmetric, otherwise the assumption of circular motion may not be appropriate. Also, to assure that our model is one describing a galaxy that ex- ists in nature, the measured rotational veloc- ities derived separately from opposite sides of the galaxy should agree to within 10%. It is also Seen to study a galaxy with an ob- served inclination between 50° and 80° so it can be verified that a disk describes the plane of the galaxy accurately all the way to the star star 28 Transactions of the Kentucky Academy of Science 57(1) edges and that the azimuthal distribution of the gas can be accurately determined. For DDO 170, all of these properties were ob- servationally verified (Lake, Schommer, and van Gorkom 1990) and this galaxy is, there- fore, exceptionally useful for our purposes. METHODS The Potential-Velocity Relationship If the galaxy is assumed to be an axisym- metric collection of gravitationally bound masses rotating about some axis, we can de- rive an expression for the relationship be- tween the gravitational potential and orbital velocity of matter in the galaxy. The result of such an exercise gives us the rotation curve. This procedure is described well in the lit- erature (eign Binney and Tremaine 1987). Because it is known that the Newtonian potential cannot be used to accurately derive the rotation curve for the galaxy DDO 170 using only a two-component mass model (Lake, Schommer, and van Gorkom 1990), we must look to modifications of the gravi- tational potential to derive an acceptable po- tential-velocity relationship. The non-New- tonian theory of gravity known as MOdified Newtonian Dynamics (MOND) (Milgrom 1983) could possibly provide us with such a modification. However, MOND has failed in its attempts to provide a solution to the ga- lactic rotation curve problem without the use of dark matter for this and several other gal- axies (Lake 1989; Lake, Schommer, and van Gorkom 1990). Another possibility lies in the framework of a modification of classical general relativity known as conformal gravity. Kazanas and Mannheim (1989) derived an exact expres- sion for the gravitational potential on all dis- tance scales in the presence of a static, spher- ically symmetric source. This solution is the analog of the Schwarzschild solution in gen- eral relativity. In conformal gravity the New- tonian potential is modified to include a lin- ear term and a quadratic term in radius. Mannheim and Kazanas (1992) argued that, Figure 1. although the Schwarzschild-like solution of the conformal gravity field equations produc- es the expected short-distance (Newtonian) potential, on larger distance scales the mod- ifications become important. These authors have used this solution up to the linear mod- ification of the potential to successfully de- scribe the rotation curves for galaxies NGC 3198, NGC 2903, NGC 5907, and DDO 154 without the use of dark matter (Mannheim 1993). These four galaxies are representative of four luminosity classes: intermediate, com- pact bright, large bright, and dwarf. Because we are particularly interested in dwarf gal- axies we note that the rotation curve for DDO 154 was successfully derived using only the visible mass. Mannheim and Kazanas (1992) noted that their treatment of DDO 154 led to some anomalous results in light of the rotation curve behavior of the other three galaxies. We also note that the fits to the ro- tation curve in conformal gravity using only the linear modification for DDO 154 indicate that the galaxy appears to be gas-rich where Mya/Moas = 5-2 and Myia/Ly = 14. Although it is interesting that conformal gravity seems to have had some success pre- dicting rotation curves by using two-compo- nent mass models, the reasons for using only the linear modification to make a prediction are unclear. Therefore, we use two methods to derive a two-component mass model for DDO 170. The first employs a modified Newtonian potential including only the linear modification, which we refer to as the “trun- cated model”; the second incorporates both linear and quadratic modifications given in Mannheim (1993) and Mannheim and Kazan- as (1992). In conformal gravity the gravitational poten- tial for a static, spherically symmetric gravitat- ing object V is given in Mannheim (1993) Vig e2B/n se yiies kre (1) where the constants B, yy, and k are integration constants depending on the mass of the grav- itating system under study. Because a galaxy is —_ Rotation curve fits for the galaxy DDO 170. The solid line is the fit to the data points when (a) the full conformal gravity model is used and (b) the linear potential model is used. The data are from Lake, Schommer, and van Gorkom (1990). velocity (km/s) velocity (km/s) Dwarf Galaxy DDO 170— Falbo-Kenkel and Kremer quadratic data ~— 20 40 60 80 100 120 140 160 radius (arcsec) 20 40 60 80 100 120 140 160 radius (arcsec) 180 30 Transactions of the Kentucky Academy of Science 57(1) a collection of stars with potential given in equation (1) we can derive the potential of the galaxy if we know the stellar and gas distri- bution functions so that the gravitational po- tential for an extended, axially symmetric disk with matter distribution given by p(R, z’) is WAGE a) = 218} | dR dd’ dz' -[Rp(R, z')] [r? + R? — 2rR cos ' rican Jae ae AY | dR dd’ dz' -Rp(R, z’)[r?+ R? =} | dR do’ dz' SR Riazi) ine chee — 2rR cos ’ +(—2'¥] (2) where the primed coordinates represent source coordinates and the unprimed coordi- nates refer to observational coordinates. The complete expression (2) was used in the full conformal gravity model while we set k = 0 in the truncated model. Given the correctness of the assumptions about DDO 170 listed above, the potential given in (2) can then be used in the well-known expression relating cir- cular velocity and potential: u(r) = [rV’(r)]*. (3) Once the matter distribution p(R,z’) for the galaxy is determined via observation, the right hand side of equation (3) can be evaluated, and a rotation curve can be derived. Several authors have been able to analytically reduce equation (2) to a more compact expression for several special cases (Binney and Tremaine 1987; Cuddeford 1993: Mannheim 1993): however for the sake of generality we chose to leave it in this form and to perform the necessary numerical integration to evaluate equation (3). The gravitational potential for every gravita- tionally bound system depends on the specific distribution of matter in the system via the function p(R, z’), which is determined by ob- servation. We used only the visible mass of stars and gas in DDO 170 to determine the exact form for the gravitational potential and used this potential to determine the rotation curve. The variable parameters in the rotation curve fits are the integration constants in the confor- mal gravity model and the ratio M,,/Moga SO that equation (3) becomes ! MT / WY v(r) ar lee ee i) at franc) 7 galaxy (4) Observed Star and Gas Mass DDO 170 is a dwarf galaxy of absolute mag- nitude M, = —15.15 with the approximate lo- cation of 13513" RA 25*42’ DEC. The galaxy is inclined at an angle of 84° and has major and minor axis lengths of 4.7 and 0.9 arcsec, respectively. The radius of the HI gas disk is about five times larger that the optical disk radius. Details of the optical and radio obser- vations as well as the details of the rotation curve derivation can be found in Lake, Schommer, and van Gorkom (1990). Here we quote only the details necessary to describe the two-component mass model and resulting rotation curve. The luminosity profile can be described as an exponential while the HI profile is de- scribed well by a Gaussian. We use these ex- pressions in equation (2) so that p(R, z’) = (M/L)e-®“f(z’) and p(R, z’) = e ®*2)f(z)) fox the HII and HI distributions, respectively. In accordance with the mass model used by Lake, Schommer, and van Gorkom (1990), the function f(z’) is taken as 1 in the case of the gas distribution and an exponential in the case of the stellar distribution. Since f(z’) has not been precisely determined by observation, we numerically studied the dependence of the ro- tation curve on f(z’) (unpublished results) and concluded that the dependence is small when f(z’) assumes a form common to that observed for other axisymmetric galaxies (Cuddeford 1993: van der Kruit and Searle 1981). RESULTS AND DISCUSSION Galactic Rotation Curve for DDO 170 Using only the visible mass and gas along with the procedure described above, we have Dwarf Galaxy DDO 170— Falbo-Kenkel and Kremer 3] derived a rotation curve for DDO 170 using two methods (Figure 1). According to our best fit, we have determined a constant mass-to- light ratio of about four for the stellar disk and derived our rotation curve using observations of R in units of arcsec and not kpc to minimize the effects of error due to measurements of distance to the galaxy. In Figure 1 we show the results of fitting our model to the data us- ing the full conformal gravity potential and truncated potential model, respectively, when the ratio M/L,,,, = 4.0 (as determined by ob- servation) and M,,/M.a. 3. This last ratio is over an order o ee aae lower than that used in three-component dark matter models; however, this is to be expected given the mod- ification of the potential. We have maximized the contribution of the stellar disk to the ro- tation curve (hence the term “maximum disk” model) in order to compare more directly the fits for this particular dwarf galaxy with rota- tion curve fits for bright spirals. It is obvious from comparison of Figure la and Figure 1b that the full conformal gravity fit models the observed rotation curve more accurately. Note that the quadratic term be- comes important far from the edge of the stel- lar disk and serves only to keep the rotation curve from rising too fast. Mannheim (1993) found the quadratic term to be negligible in the rotation curve fits and suggested that the quadratic term becomes important only on cosmological distance scales. Here, however, we find that k,,),., = 3.6 X 10-°°cm~ and has a noticeable affect on the rotation curve be- ginning at about three times the radius of the stellar disk. It is also true that the linear term (y = 8.4 X 10°%8cm~!) works well to describe the rotation curve up to that point. These results lead us to question the phys- ical interpretation of k and the quadratic term as cosmological. To further address this issue, we plan to analyze the rotation curve of an- other dwarf galaxy, DDO 154, using the full conformal gravity potential to see if the “anomalous” behavior refered to be Mann- heim (1993) might be accounted for in this way, or if this behavior is truly an indicator of some basic physical property of low luminosity (dwarf) galaxies. SUMMARY We have shown that it is not necessary to employ dark matter as part of a three-com- ponent model to derive the rotation curve for the dwarf galaxy DDO 170. By use of the full conformal gravity potential, the measured ro- tation curve for the dwarf galaxy DDO 170 can be derived using a two-component mass model that consists of (1) a luminous disk with constant mass-to-light ratio and (2) an HI gas disk. A good fit to the measured rotation curve can be obtained when a “maximal disk” model is employed. To understand whether or not two-compo- nent models can in general be used to under- stand the mass distribution in other dwarf gal- axies as well as spiral galaxies, the procedure employed here to deteudane the rotation curve will continue to be used. Future studies will help us to better understand (1) how the integration constants in conformal gravity scale with mass, (2) overall trends regarding the distribution of visible stars and gas in bright spiral galaxies as opposed to dwarf spi- ral and dwarf irregular galaxies, and (3) the dependence of the ratio M,,/M,,, on mor- phology. a ss ACKNOWLEDGEMENTS We thank Demos Kazanas and Phillip Mannheim for many illuminating discussions and the Laboratory for High Energy Astro- physics at Goddard Space Flight Center for its hospitality in summer 1993. Computing sup- port from Kristin Pfabe and the Department of Physics at the University of Cincinnati is also acknowledged and greatly appreciated. This research was supported by the Office of Research, Grants, and Contracts and the De- partment of Physics and Geology at Northern Kentucky University under NASA agreement JOVE-O01T-1-01-1900-4351. LITERATURE CITED Binney, J., and S. Tremaine. 1987. Galactic dynamics. Princeton University Press, Princeton, NJ. Broeils, A. H. 1992. The mass distribution of the dwarf spiral NGC 1560. Astron. Astrophys. 256:19-32. Carignan, C., and S. Beaulieu. 1989. Optical and HI studies of the “gas-rich” dwarf irregular galaxy DDO 154. Astrophys. J. 347:760-770. Cuddeford, P. 1993. On the potentials of galactic discs. Monthly Notices Roy. Astron. Soc. 262:1076—1086. Kazanas, D., and P. D. Mannheim. 1989. Exact vacuum solution to conformal Weyl gravity and galactic rotation curves. Astrophys. J. 342:635-637. 32 Transactions of the Kentucky Academy of Science 57(1) Lake, G. 1989. Testing modifications of gravity. Astro- phys. J. 345:L17-L19. Lake, G., R. A. Schommer, and J. H. van Gorkom. 1990. The distribution of dark matter in the dwarf galaxy DDO 170. Astron. J. 99(2):547—560. Mannheim, P. D. 1993. Linear potentials and galactic ro- tation curves. Astrophys. J. 419:150-154. Mannheim, P. D., and D. Kazanas. 1992. Newtonian limit of conformal gravity and lack of necessity of the second order Poisson equation. Gen. Relativ. Gravitat. 26(4): 337-361. Milgrom, M. 1983. Modified Newtonian dynamics. As- trophys. J. 270:365-371, 384. van der Kruit, P. C., and L. Searle. 1981. Surface pho- tometry of edge-on spiral galaxies. Astron. Astrophys. 95:105-115. Trans. Ky. Acad. Sci. 57(1):33-46. 1996. Unionid Fauna of the Lower Cumberland River from Barkley Dam to the Ohio River, Kentucky (Mollusca: Bivalvia: Unionidae) James B. Sickel Department of Biological Sciences, Murray State University, Murray, Kentucky 42071 and Carol C. Chandler Natural Resources Conservation Service, U.S. Department of Agriculture, 286 East Main Street, Gallatin, Tennessee 37066 ABSTRACT Unionids of the lower Cumberland River, Kentucky, were surveyed by brail and SCUBA from Barkley Dam to the Ohio River. The only previous survey was conducted in 1910-1911 by Wilson and Clark. The two surveys are compared and show a significant decrease in the number of surviving species. Live individuals of 24 species were found in our surveys compared to 45 species found by Wilson and Clark in the region now inundated by Lake Barkley. Three species were found that had not been reported previously from the Cumberland River. Alteration of habitat resulting from impoundment, pollution, and rapid fluctuation of discharge from hydroelectric dams apparently are the major causes for the decline in native species, while impoundment probably has enhanced invasion by species from the Ohio River. INTRODUCTION In recent years the number of species of unionids on the federal endangered species list (USFWS 1993) has increased, and there is growing interest in the scientific community in conserving biodiversity and understanding its importance to a healthy environment (Wilson 1994). In contrast, there is an apparent waning interest in protecting rare, threatened, and en- dangered species and the environment by the current U.S. Congress. With the rapid spread of zebra mussels into the Ohio and Mississippi river systems, an already impoverished native unionid fauna is faced with greater threats, making it imperative that unique, rare, and endangered species be studied and docu- mented. Documentation of species and recent changes in their abundance and distribution is critical for understanding the importance of complex ecosystems if biodiversity is to be preserved. With these issues in mind, the re- sults of several surveys of the lower Cumber- land River conducted by us in 1981 and 1988 are reported here and compared to a 1910— 1911 survey by Wilson and Clark (1914). STUDY AREA The Cumberland River arises in the Cum- berland Mountains in southeastern Kentucky. 33 It flows westerly, arching southward into cen- tral Tennessee. From there it flows westerly, gradually arching northward into western Kentucky and northwesterly to its junction with the Ohio River at Smithland, Kentucky. The “official” U.S. Army Corps of Engineers junction, Cumberland River mile (CRM) 0.0, occurs at Ohio River mile (ORM) (Anonymous 1994). The actual confluence of the two bodies of water where mixing poe is at the official Cumberland River mile 2 (Figure 1). The Cumberland River and me larger and nearly parallel Tennessee River are two of the most ancient rivers of North Amer- ica. This in part is the reason for the great diversity of unionids that once populated the rivers (Wilson and Clark 1914). Documenta- tion of the unionids of the middle and upper Cumberland River by numerous authors in re- cent years indicates major declines of the unionid fauna of those areas (Anderson, Lay- zer, and Gordon 1991; Blankenship and Crockett 1972; Call and Parmalee 1982; Clark 1981, 1985: Harker et al. 1980; Isom, Gooch, and Dennis 1979; Miller, Rhodes, and Tippit 1984: Neel and Allen 1964; Parmalee, Klippel and Bogan 1980; Schmidt 1982; Schuster, But- ler, and Stansbery 1989; Starnes and Bogan 1982). Two important reviews providing gen- 922-7. 34 Bissel Q Vicksburg Hickory Creek 1 0 1 Smith Scale in Miles Creek Ferguson Creek McCormick Q Smithland Creek Section of lower Cumberland River shown in detail. KENTUCKY 25025150, ---_———__] BARKLEY DAM Scale in Miles LAKE BARKLEY Figure 1. eral information about unionid distribution in the Cumberland River are those by Cicerello, Warren, and Schuster (1991) for Kentucky and Starnes and Bogan (1988) for Tennessee. WILSON AND CLARK 1910-1911 SURVEY Charles B. Wilson and H. Walton Clark (1914) published the results of a freshwater unionid survey of the Cumberland River con- ducted by them, John F. Boepple, and Ernest Danglade and supervised by Robert E. Coker. Their project involved several expeditions in summers of 1910 and 1911 to evaluate the shell resources then being utilized in the pearl button industry. By sampling with crowfoot Transactions of the Kentucky Academy of Science 57(1) LIVINGSTON COUNTY CRITTENDEN ce Pinckneyville -"" COUNTY a} * Claylick .f Creek 3 17 16 Sugar Cypress Creek ue Creek LIVINGSTON COUNTY by cosbarg 23 Creek Richland Creek | LYON COUNTY Feinklestein Branch Cook Big Crevasse Creek Branch |-24 7 28 Nokes Creek Flat Creek Barkley Dam KN Lake Barkley Map of lower Cumberland River, Kentucky, from Barkley Dam at Cumberland River mile 30.6 to the Ohio River showing major unionid beds as elongated, solid areas and river mile points as solid dots. Numbers 2 through 30 along the river represent miles from its confluence with the Ohio River. (Width of river is shown as twice actual scale.) brails, tongs, and rakes in deep water, wading in shallow water, and examining the catch of commercial shellers, they described the spe- cies composition and relative abundance of unionids from the headwaters above Cumber- land Falls in eastern Kentucky to near the mouth where the Cumberland River enters the Ohio River. Their sample location farthest downstream was at Horse Ford near Kuttawa, Lyon County, Kentucky, at Cumberland River mile (CRM) 36.5 (58.7 km from the Ohio Riv- er at Smithland, Livingston County, Ken- tucky). They reported extensive beds from Cumberland Falls to Kuttawa. That study con- stitutes the only published report prior to our Unionid Fauna of Lower Cumberland River, surveys of the unionids inhabiting the lower Cumberland River, including the region pres- ently inundated by Lake Barkley. In their re- port, Wilson and Clark (1914) briefly dis- cussed ecological requirements of some unionids and the influence of dams on this fauna. The effect of dams on unionids living in once free-flowing rivers has been extensive (Bates 1962; Fuller 1974). When Wilson and Clark (1914) were conducting their survey, only one dam had been constructed on the lower Cumberland River below Nashville, Tennessee; eight had been completed up- stream from Nashville. Lock and Dam A at CRM 150.6 were completed by the Army Corps of Engineers in 1904 near the present site of @heatham Dam (CRM 148.7), Chea- tham County, Tennessee, which was complet- ed in 1957. Cheatham Dam forms the up- stream boundary of Lake Barkley. Dam A formed a pool that backed up the Harpeth River entering at CRM 153. Wilson and Clark (1914) reported that siltation in the lower Harpeth River resulting from Dam A had eliminated a large unionid bed. Similar results were observed immediately upstream from other dams. However, Wilson and Clark re- ported rapid growth of unionids in the back- waters above the silt, and they believed that the dams would be more beneficial than harmful. They were referring to low dams, however, that did not raise the water above the original river banks. They had no experi- ence with the high dams such as Barkley and Cheatham dams presently on the lower Cum- berland River. From near the present location of Chea- tham Dam (CRM 148.7) to Barkley Dam (CRM 30.6), Lyon County, Kentucky, Wilson and Clark (1914) collected at 28 stations. From their descriptions many of their stations can be located on current maps or navigation charts. In the following discussion, locations given as approximate river miles could not be located precisely. Below mile 148, the up- stream boundary of the present Lake Barkley, Wilson and Clark reported 47 species (Table 1). Eleven of the 47 species are listed on the federal endangered species list (USFWS 1993): Cyprogenia stegaria, Dromus dromas, Epioblasma florentina, E. haysiana, E. obli- quata, Hemistena lata, Lampsilis abrupta, Kentucky—Sickel and Chandler Bi) Obovaria retusa, Plethobasus cooperianus, Pleurobema plenum, and Quadrula_ fragosa. Cyprogenia stegaria was reported at Seven- mile Ferry (CRM 132.5), Owl Hollow Bar (ca. CRM 130), and Guisers [Geisers| bar (CRM 128.5). Dromus dromas occurred at Seven- mile Ferry and Red Rock Bar (ca. CRM 125) below Clarksville, Montgomery County, Ten- nessee. Only a shell of Epioblasma florentina was found at Half Pone Bar (CRM 145). Epioblasma haysiana was found at Clarksville (CRM 126) and above Ball [Bald] Island (CRM 97.6). Epioblasma obliquata was re- ported alive only from Half Pone Bar; a dead shell was found at Elk Creek Shoals (CRM 98.6). Hemistena lata was reported from Half Pone Bar. Lampsilis abrupta was reported from Seven-mile Ferry and Kuttawa, Ken- tucky (CRM 41). Obovaria retusa was report- ed from above Ball Island, foot of Dover Is- land (CRM 90.5), Canton, Kentucky (CRM 62), and Horse Ford (CRM 36.6). Plethobasus cooperianus was reported from Owl Hollow Bar, Guisers Bar, Clarksville, Red Rock Bar, Meeks Spring Bar (ca. CRM 118), Walters’ Camp (ca. CRM 98), above and below Ball Island (CRM 97.4), Linton (CRM 73), Do- nelson [Donaldson] Creek (CRM 67), Canton (CRM 63), and Horse Ford below Kuttawa. At Walters’ shell camp in 1911 Wilson and Clark noted a pile of ca. 150 tons of shells with Plethobasus cooperianus ranking second in abundance of the commercial species. Pleu- robema plenum was found at Half Pone Bar, Seven-mile Ferry, and Jones Landing (ca. CRM 80) somewhere between Dover, Stewart County, Tennessee, and Linton, Trigg County, Kentucky. Quadrula fragosa was reported from Half Pone Bar, Seven-mile Ferry, Owl Hollow Bar, Clarksville, Red Rock Bar, Trices Landing (ca. CRM 124), above Meeks Spring Bar (ca. CRM 119), Meeks Spring Bar, foot of Dover Island, Jones Landing, Linton, below Canton, Eddyville Bar (CRM 44), above Kut- tawa, below Money Cliff (CRM 39), and at Horse Ford. Wilson and Clark did not collect downstream from Horse Ford at CRM 36.5. Therefore, no records exist before our 1981 and 1988 surveys of the unionid fauna in the region now the tailwater of Barkley Dam from CRM 30.6 to the Ohio River. Historically, the Cumberland River con- tained at least 85 species of unionid bivalves 36 Transactions of the Kentucky Academy of Science 57(1) Table 1. Unionid clams of the lower Cumberland River, Kentucky, from the region now inundated by Barkley Lake to the Ohio River as reported by Wilson and Clark (1914) from CRM 36 - 148 and surveys in 1981 and 1988 from CRM 2.5 - 47. Taxa! Actinonaias ligamentina (Lamarck, 1819) Amblema plicata perplicata (Conrad, 1841) Amblema plicata plicata (Say, 1817) Anodonta suborbiculata Say, 1831 Arcidens confragosus (Say, 1829) Cumberlandia monodonta (Say, 1829) Cyclonaias tuberculata (Rafinesque, 1820) Cyclonaias tuberculata (Rafinesque, 1820) Cyprogenia stegaria (Rafinesque, 1820) Dromus dromas (Lea, 1834) Ellipsaria lineolata (Rafinesque, 1820) Elliptio crassidens (Lamarck, 1819) Elliptio dilatata (Rafinesque, 1820) Epioblasma flexuosa (Raf., 1820) Epioblasma florentina florentina (Lea, 1857) Epioblasma haysiana (Lea, 1833) Epioblasma obliquata (Rafinesque, 1820) Fusconaia ebena (Lea, 1831) Fusconaia flava (Rafinesque, 1820) Fusconaia subrotunda (Lea, 1831) Hemistena lata (Rafinesque, 1820) Lampsilis abrupta (Say, 1831) Lampsilis ovata (Say, 1817) Lampsilis ventricosa (Barnes, 1823) Lampsilis anodontoides (Lea, 1831) Lampsilis teres (Rafinesque, 1820) Lasmigona complanata (Barnes, 1823) Leptodea fragilis (Rafinesque, 1820) Ligumia recta (Lamarck, 1819) Megalonaias nervosa (Rafinesque, 1820) Obliquaria reflexa Rafinesque, 1820 Obovaria olivaria (Rafinesque, 1820) Obovaria retusa (Lamarck, 1819) Obovaria subrotunda (Rafinesque, 1820) Plethobasus cicatricosus (Say, 1829) Plethobasus cooperianus (Lea, 1834) Plethobasus cyphyus (Rafinesque, 1820) Pleurobema catillus (Conrad, 1836)" Common Name Mucket Roundlake Threeridge Flat floater Rock pocketbook Spectaclecase Purple wartyback Purple wartyback Fanshell Dromedary mussel Butterfly Elephant-ear Spike Leafshell Yellow blossom Acornshell Catspaw Ebonyshell Wabash pigtoe Long solid Cracking pearly Pink mucket Pocketbook Pocketbook Yellow sandshell Yellow sandshell White heelsplitter Fragile papershell Black sandshell Washboard Threehorn wartyback Hickorynut Ring pink Round hickorynut White wartyback Orangefoot pimpleback Sheepnose Solid pigtoe Reported Present Nomenclature of Wilson and Clark (1914) in 1914 Surveys Lampsilis ligamentina gibba (Simpson, 1900) Quadrula perplicata (Conrad, 1841) Quadrula undulata (Barnes, 1823) Margaritana monodonta (Say, 1829) Quadrula tuberculata (Rafinesque, 1820) Quadrula granifera (Lea, 1838) Cyprogenia irrorata (Lea, 1828) Dromus dromas (Lea, 1834) Plagiola securis (Lea, 1829) Unio crassidens crassidens Lamarck, 1819 Unio gibbosus Barnes, 1823 Truncilla florentina (Lea, 1857) Truncilla haysiana (Lea, 1834) Truncilla sulcata (Lea, 1829) Quadrula ebena (Lea, 1831) Quadrula undata (Bames, 1823) Quadrula subrotunda (Lea, 1831) Lastena lata (Rafinesque, 1820) Lampsilis orbiculata (Hildreth, 1828) Lampsilis ovata (Say, 1817) Lampsilis ventricosa (Bames, 1823) Lampsilis anodontoides (Lea, 1831) Lampsilis fallaciosa (Smith, 1899) Symphynota complanata (Barnes, 1823) Lampsilis gracilis (Barnes, 1823) Lampsilis recta (Lamarck, 1819) Quadrula heros (Say, 1829) Obliquaria reflexa Rafinesque, 1820 Obovaria ellipsis (Lea, 1828) Obovaria retusa (Lamarck, 1819) Obovaria circulus (Lea, 1829) Quadrula cooperiana (Lea, 1834) Pleurobema aesopus (Green, 1827) Quadrula solida (Lea, 1838) Unionid Fauna of Lower Cumberland River, Table 1. (continued) Taxa! Common Name Pleurobema cordatum (Rafinesque, 1820) Ohio pigtoe Pleurobema plenum (Lea, 1840) Rough pigtoe Pleurobema rubrum (Rafinesque, 1820) Pink pigtoe Pleurobema sintoxia (Rafinseque, 1820) Round pigtoe Potamilus alatus (Say, 1817) Pink heelsplitter Potamilus ohiensis (Rafinesque, 1820) Pink papershell Ptychobranchus fasciolaris (Rafinesque, 1820) Kidneyshell Pyganodon grandis (Say, 1829)? Giant floater Quadrula cylindrica cylindrica (Say, 1817) Rabbitsfoot Quadrula fragosa (Conrad, 1835) Winged mapleleaf Monkeyface Wartyback Mapleleaf Quadrula metanevra (Rafinesque, 1820) Quadrula nodulata (Rafinesque, 1820) Quadrula quadrula (Rafinesque, 1820) Quadrula pustulosa pustulosa (Lea, 1831) Pimpleback Strophitus undulatus undulatus (Say, 1817) | Squawfoot Tritogonia verrucosa (Rafinesque, 1820) Pistolgrip Truncilla donaciformis (Lea, 1828) Fawnsfoot Truncilla truncata Rafinesque, 1820 Deertoe Utterbackia imbecillis (Say, 1829) Villosa lienosa (Conrad, 1834) Paper pondshell Little spectaclecase Kentucky—Sickel and Chandler aT Present Reported in 1914 Nomenclature of Wilson and Clark Surveys Quadrula obliqua (Lamarck, 1819) Quadrula plena (Lea, 1840) Quadrula pyramidatum (Lea, 1831) Quadrula coccinea (Conrad, 1834) Lampsilis alata (Say, 1817) Lampsilis laevissima (Lea, 1829) Ptychobranchus phaseolus (Hildreth, 1828) Anodonta grandis Say, 1829 Quadrula cylindrica (Say, 1817) Quadrula fragosa (Rafinesque, 1820) Quadrula metanevra (Rafinesque, 1820) Quadrula lachrymosa (Lea, 1828) Quadrula pustulosa (Lea, 1831) Strophitus edentulus shaefferiana (Lea, 1852) Quadrula tritogonia Ortmann, 1909 Plagiola donaciformis (Lea, 1828) Plagiola elegans (Lea, 1831) Anodonta imbecillis Say, 1829 Lampsilis lienosa (Conrad, 1834) (Hyphen indicates not found, plus indicates found alive, and “shell” indicates found only as a dead shell) "Nomenclature agrees with that of Turgeon et al. (1988) except where noted otherwise ? Nomenclature of Ortmann and Walker (1922) 3 Hoeh (1990) (Starnes and Bogan 1988). This number is sec- ond only to the Tennessee River, which once had ca. 95 species (Starnes and Bogan 1988), the largest assemblage found anywhere (John- son 1978). Many species once found in the middle and upper reaches of the Cumberland River belong to the Cumberlandian fauna, those unionids that apparently had their origin in the Cumberland and Tennessee river sys- tems. Ortmann (1924) defined the Cumber- landian Region as extending from the head- waters of the Cumberland River down to near Clarksville, Tennessee, and the headwaters of the Tennessee River down to some point be- yond Muscle Shoals, Alabama, including part of the Duck River, Tennessee. Of the species reported by Wilson and Clark (1914) in the lower Cumberland River, only three were considered to be Cumberlan- dian forms (origin in Cumberlandian Region) by Ortmann (1925): Dromus dromas, Epiob- lasma haysiana, and E. florentina. Johnson (1980), however, added three more to the list, claiming that they had extended their range from the Cumberlandian Region into the Ohio drainage: Epioblasma flexuosa, Lampsilis abrupta, and Plethobasus cicatricosus. In the case of Lampsilis abrupta, Ortmann (1925) stated that it is a large river form and its “cen- ter of origin is in the Interior Basin.” With the demise of the pearl button indus- try, the lower Cumberland River saw few shellers. When the demand for shells for the cultured pearl industry sent shellers back to the rivers, the discovery that Cumberland Riv- er shells from below Nashville were too chalky or badly eroded and had a low value for the production of pearl nuclei kept shellers away. 38 Transactions of the Kentucky Academy of Science 57(1) Therefore, little was known about the recent unionid fauna until our surveys were conduct- ed. The purpose of our study was to determine the distribution and relative abundance of the unionid fauna of the lower Cumberland River from Barkley Dam to the Ohio River. The Nashville District Corps of Engineers had pro- posed channel maintenance operations that potentially could have an adverse impact on the mollusks. The Corps was required to as- certain whether any species on the federal en- dangered species list survived in the area of the river being considered for channel im- provement. We present results of that survey and a comparison with the Wilson and Clark (1914) study along with a report on the range extension of three species into the Cumber- land River and a comparison with archeologi- cal studies. 1981 AND 1988 SURVEYS Methods The 1981 survey was conducted using three commercial boats equipped with 4.9 m crow- foot brails and operated by commercial shell- ers. Brails have been used in the shell industry since the end of the 19th century (Coker 1919). Our brails consisted of a metal or wooden bar with 1 m lengths of line or light chain attached every 7.6 cm. Attached to the lines were bundles of four-pronged hooks made of heavy (10 to 14 gauge) wire. The ends of the wire were bent upward at various angles depending on the type of sediment in which the brail was used and on the method of haul- ing. The tips of the hooks were melted to form a bead to help hold the unionid, a modification invented by J. F. Boepple (Wilson and Clark 1914), one of the founders of the pearl button industry in the United States. One or two brails were towed slowly by a boat such that the bar was suspended just above the sedi- ment surface and the hooks raked the bottom. A hook tip that enters the opening between the two valves of a unionid will cause the in- dividual to clamp shut on the hook. If the hook is secure it will pull the individual from the sediment; the individual will remain at- tached until the brail is hauled to the surface. A jonboat equipped with a 2.4 m brail and operated by Murray State University person- nel was also used. Three SCUBA divers sam- pled selected sites where brails were not ef- fective, such as among large rocks and snags. Shorelines were searched for empty shells that may have washed ashore or piles of shells left by muskrats. Both conditions usually indicate that a bed is nearby. Beds discovered by brail- ing and areas in the vicinity of shell piles ob- served along the shoreline were also examined by divers. The commercial shellers brailed the middle, right, and left margins of the channel from river mile 30 to the Ohio River at CRM 2.2. Where unionids were encountered in numbers suggesting a bed, several brail hauls were made to more fully determine the loca- tion and extent of the bed and the species composition. SCUBA diving on beds was used to search for species not caught by the brails. Representative samples of brail catches and all SCUBA samples were counted to determine the relative abundance of species. Other brail samples were examined only for species com- position. Empty shells found along shore were examined for species not encountered by oth- er collecting methods. The 1988 survey was conducted by two SCUBA divers. One addi- tional site was sampled by divers in Lake Bark- ley at Clay Bay, CRM 47.0. Names of species correspond to those given in Turgeon et al. (1988) except where otherwise noted. Results and Discussion The distribution of all species found during our 1981 and 1988 surveys is presented in Ta- ble 2. The sample locations are reported by river mile and position within the river as ei- ther left, middle, or right of the channel as viewed facing downstream. Unionid beds. Unionid beds are considered to be locations of stable substrate, usually of gravel and sand stabilized by compact silt and clay, in which individuals of various age classes and species occur in significant densities, gen- erally more than 1/m?. The establishment of a bed requires many years since recruitment is generally a slow process. It is not uncommon to find beds composed of individuals ranging from 5 to 25 years in age with very few juve- niles. Figure 1 shows the Cumberland River from Barkley Dam to the Ohio River. Elongated solid areas indicate the major beds found in the surveys. The small, oval, solid areas indi- Unionid Fauna of Lower Cumberland River, Kentucky cate smaller concentrations. The only bed that several retired commercial shellers could re- call as ever having produced commercial har- vests was in the vicinity of mile 14 between Pinckneyville and Sandy Creek. Diving and brailing at the site revealed a bed with limits from mid-channel to the right bank from CRM 13.1—14.7. Nineteen species were found in the bed, Stich was the largest and most species rich in the Barkley Dam tailwater sec- tion of the river. The age composition of the bed indicated that some of the species in this section of the river may be on the verge of demise, most individuals being over 15 years old with no evidence of recruitment since the construction of Barkley Dam in 1965. The rapid rise and fall of water level caused by the intermittent discharge of water for power gen- eration at Barkley Dam may be one cause for the lack of recruitment. The surveys located other beds at CRM 4.5-5.0, middle; CRM 9.4—11.0, middle; CRM 17.0-17.3, right-middle-left; and CRM 26.5- 27.2, middle-right. The bed at CRM 4.5-5.0, confined to a narrow mid-channel region, con- sisted mainly of Megalonaias nervosa with mi- nor representatives of 13 other species. The bed at CRM 9.4—11.0 was rather spread out down the mid-channel. Fourteen species were represented with Fusconaia ebena being the most abundant and Megalonaias nervosa, El- liptio crassidens, and Pleurobema cordatum following in that order. At CRM 17.0-17.3 a small bed was located in the vicinity of another old shell pile on the right bank at CRM 17.3. The bed was small but extended from bank to bank and con- tained 12 species. The bed at CRM 26.5-27.1 was dense and extended from the bottom of the right bank out to mid-channel. Fifteen species were re- covered alive along with empty shells of Obo- varia retusa and Pleurobema rubrum. This bed is in danger of being destroyed by channel maintenance dredging. Old shell piles along shore. Because of the steep river banks tice bank slumping caused by the fluctuating discharge at Barkley Dam and wave action from passing vessels, there were few shoreline sites where empty shells were likely to wash ashore and few locations where old shell piles remained exposed. Only a few shells were found on gravel bars or clay 39 banks; most of these were Corbicula fluminea (Miiller), one of the Asiatic clams. Three large shell piles were found mostly buried in the bank, which had apparently slumped over them. These were located on the right bank at miles 14.3, 17.3, and 19.5. Digging in the bank uncovered many shells, mostly “pinks,” Cyclonaias tuberculata, Ellip- tio crassidens, and Elliptio dilatata, and “washboards,” Megalonaias nervosa. “Pinks” are shells with a pink or purple nacre that were not used in the pearl button industry; they were usually culled by shellers along shore to remove them from the harvest and to prevent them from returning to the beds. Washboards in the Cumberland River were generally stained and of low value, so they too were frequently culled (Wilson and Clark 1914). Because of the species composition of the piles, it is assumed that they are old cull piles from pearl button days 40 to 90 years ago. Commercial shellers probably culled the unwanted shells along shore near the beds where they were working. Two of the three shell piles occurred adjacent to existing beds, which supports the idea that the beds are very old. Some of the living individuals in these beds could well be older than 30 years. It is difficult to determine accurately the age of a clam beyond about 15 years without thin sec- tioning the shell because the shell growth slows and the external rest lines fence to be poorly separated. The third pile was near a landing at Dycusburg, probably a location where commercial shellers processed their harvest. Comparison with Wilson and Clark survey. All of the unionids found in our study and those reported by Wilson and Clark (1914) for the lower Cumberland River are listed in Ta- ble 1. Wilson and Clark reported 45 species from between CRM 148-125 and 33 species from CRM 124-36. Our survey lists 33 species (24 found alive) from CRM 47-2.2. Of the 14 species Wilson and Clark reported from above CRM 124 that did not occur below that point, two are Cumberlandian forms (Ortmann 1925): Dromus dromas and Epioblasma flo- rentina. One Cumberlandian species, Epio- blasma haysiana, was reported by Wilson and Clark below CRM 124 from above Ball Island at CRM 98. It is unlikely that these species still exist in the lower Cumberland River. Ac- 40 Transactions of the Kentucky Academy of Science 57(1) Table 2. Abundance of live unionid clams collected by brailing or SCUBA diving left, middle, or right of the river channel in surveys conducted in 1981 (a) and 1988 (b) in the lower Cumberland River, Kentucky, from CRM 47 to the Ohio River. (The letter S indicates that only one or more dead shells were found.) ee Middle, Brail (a) Middle, Brail (a) CRM 10.6-11.0 Middle, Brail (a) ¥ CRM 11.0-12.4 Middle, Brail (a) CRM 13.5-14.0 Right, Brail (a) Right, Dive (a) Right, Dive (a) CRM 4.7-5.0 CRM 4.9 CRM 9.4-11.0 Left, Dive (a) CRM 14.2 Taxa ” |Left, Dive (b) Amblema plicata ace Epioblasma flexuosa Fusconaia ebena Lampsilis abrupta — oS Lasmigona complanata Plethobasus cicatricosus Plethobasus cooperianus Pleurobema sintoxia oS Pleurobema cordatum Truncilla donaciformis Truncilla truncata TOTAL LIVE UNIONIDS | 106 TOTAL SPECIES LIVE 12 sa ia = nl pide ee i ay Ee N aE Unionid Fauna of Lower Cumberland River, Kentucky—Sickel and Chandler 41 Table 2. (continued) “=~ N + S&S = fan . = Eh Reo — C) 2 E~ o S& Cisse = =~ es) — — — o > & C= TiS Ss = = Sa fe 2/S oe os Z2IS GAIA o /% = ~ A Anes Sm fa) Ge Ge llsse 6 ses — AB Q A G|AB!A + AG s re =| +s = = aS = = = elss “ob “ob 7 “ob “ob S lp "ob O m4 wil Ou O POL eno ones Amblema plicata Anodonta suborbiculata Arcidens confragosus Cyclonaias tuberculata = Ellipsaria lineolata 3 a Elliptio dilatata ae Fusconaia ebena Fusconaia flava Lampsilis abrupta Lasmigona complanata Leptodea fragilis Ligumia recta Megalonaias nervosa ed Obliquaria reflexa Obovaria retusa ethobasus cicatricosus ac fas) v |e > is) I~) : (g S i) ay 5 v o 5 g => : K| = S x Sy eames ee ues e et Ee ace tN wi = i a ff : L oo y : 2 = g = im) Ww DS WF Ie S = &. 18 1§ £8 |F s is is Le ar la a | Quadrulanodulata | Quadrula p. pustulosa | Quadrula quadrula__| | aaa ae tS tS tO ee ee TOTAL LVE UNIONIDS | 56 TOTAL SPECIES LIVE Nn Ww — 4 7 42 Transactions of the Kentucky Academy of Science 57(1) Table 2. (continued) Amblema plicata ‘Anodonta suborbiculata Middle, Brail (a) Right, Dive (a) CRM 28.5 CRM 47.0 Arcidens confragosus Cyclonaias tuberculata CRM 28.3 ~ |Right, Dive (b) — Ellipsaria lineolata Elliptio crassidens > |Left, Dive (b) Elliptio dilatata Epioblasma flexuosa 0.00 Fusconaia ebena Lampsilis abrupta Lasmigona complanata 8.06 Leptodea fragilis Ligumia recta Obliquaria reflexa Obovaria retusa 1.61 0.00 _| Plethobasus cicatricosus Plethobasus cooperianus Pleurobema sintoxia Pleurobema cordatum Pyganodon grandis 4.30 0.00 Quadrula metanevra Quadrula nodulata Quadrula p. pustulosa Quadrula quadrula Tritogonia verrucosa Truncilla donaciformis Truncilla truncata Utterbackia imbecillis 0 4.95 0.00 | 4 | 04s | TOTAL LIVE UNIONIDS TOTAL SPECIES LIVE 0.22 0.11 a aire isl Tal esa [18 lated | a 40 0 46 0 4 16 4 95 1 2 1 29 24 39 99.89 3 Unionid Fauna of Lower Cumberland River, cording to Stansbery (1970) Epioblasma _flo- rentina is restricted to the South Fork Holston River, Virginia, while Epioblasma_ florentina walkeri, perhaps the same as Epioblasma flo- rentina of Wilson and Clark, is “reduced to the lower Stones and Red Rivers of the Cumber- land River system,” and Epioblasma haysiana is restricted to a 16 km region of the Clinch River. Dromus dromas was recently found liv- ing in the Cumberland River 160 km ee stream from Nashville at CRM 296.8 by Par- malee et al. (1980), and there is a possibility that it still exists in the upstream section of Lake Barkley above Clarksville within the original Cumberlandian Region where Lake Barkley still retains some of the characteristics of a free-flowing river. Within the region now in Lake Barkley, the data from Wilson and Clark (1914) indicate a gradual change in species composition with the number of species declining downstream. This is to be expected because, as the river increases in size and decreases in gradient, the variability of habitats is reduced and fewer shoals occur, the shoals being a favored habitat for small river forms. The 14 species Wilson and Clark (1914) re- ported only from above CRM 124 (CRM 124— 148) were Cyclonaias tuberculata, Cyprogenia stegaria, Dromus dromas, Epioblasma floren- tina, E. obliquata, Fusconaia subrotunda, Hemistena lata, Lasmigona complanata, Obovaria subrotunda, Blearobionia sintoxia, Ptychobranchus fasciolaris, Truncilla donaci- formis, T. truncata, and Villosa lienosa. With the exception of the two Cumberlandian spe- cies already mentioned (Dromus dromas, Epioblasma florentina) that were not reported below CRM 125, the other 12 species are widely distributed in other river systems, and three (Lasmigona complanata, Truncilla don- aciformis, T. truncata) are present in the lower Cumberland today. Several species were found in our study that were not reported by Wilson and Clark (1914) in the lower Cumberland River and several that have never been reported from the Cum- berland River. Those not reported by Wilson and Clark are Anodonta suborbiculata, Arci- dens confragosus, Epioblasma flexuosa, Pletho- basus cicatricosus, Pleurobema rubrum, Py- ganodon grandis, Quadrula nodulata, and Ut- terbackia imbecillis. Kentucky—Sickel and Chandler 43 Epioblasma flexuosa was found in our sur- vey at CRM 27.5 only as a single valve with part of the periostracum attached. Casey (1987) reported shells from archaeological sites at CRM 26L, 26.6R, and 27.3L that date between 1000 and 1300 A.D. Neel and Allen (1964) reported Epioblasma lewisi (Walker) from the upper Cumberland River where Volf Creek Dam is now located. Both are as- sumed to be extinct (Stansbery 1970). Plethobasus cicatricosus was recovered as a single, badly eroded valve at CRM 27.6. It has not been reported live from the Cumberland River but was previously reported from pre- historic rock shelters along the middle Cum- berland River by Parmalee, Klippel, and Bo- gan (1980) and by Casey (1987) at CRM 26.6R and CRM 27.3L. Stansbery (1970) considered it restricted to a single population in the Ten- nessee River in northern Alabama below Wil- son Dam. Pleurobema rubrum was recovered only as a relic shell, both valves with most of the peri- ostracum attached, at CRM 26.9. It was not reported by Wilson and Clark (1914) from the lower Cumberland, but they found a few in the upper Cumberland. It was found by Par- malee, Klippel, and Bogan (1980) in the pre- historic rock shelter deposits along the Cum- berland River in Smith County, Tennessee. Pyganodon grandis and Utterbackia imbe- cillis were not reported by Wilson and Clark from the main river, but they were found in a floodplain lake, Haynes Lake, a short distance downstream from Clarksville, Tennessee. Three species found in our surveys were not reported from the Cumberland River by Wil- son and Clark (1914), Neel and Allen (1964), or Parmalee, Klippel, and Bogan (1980): An- odonta suborbiculata, Arcidens confragosus, and Quadrula nodulata. All three were dlisted by Johnson (1980) as having originated in the Mississippian Region. He suggested that Ar- cidens confragosus had a refugium in the Mer- amec River, Missouri, during the Pleistocene and that all three found a refuge in the Ozar- kian Region below the Ozark Crest. Johnson (1980) presumed that their extension into the Ohioan Region occurred after the Pleistocene; however he did suggest that the Green River, Kentucky, may have been a refugium during the Pleistocene. These three species appar- ently have entered the lover Cumberland Riv- 44 Transactions of the Kentucky Academy of Science 57(1) er since the Wilson and Clark survey, and the altered habitat caused by dams may have in- fluenced the invasion. In 1929 the Army Corps of Engineers completed Lock and Dam 52 at Paducah, Kentucky. This dam maintained an upstream elevation of 92 m above mean sea level. This reduced the flow in the lower Cum- berland River and may have encouraged the invasion by the three Mississippian species. The completion of Barkley Dam in 1965 may have contributed to the invasion as well. The impoundment conditions apparently have been favorable for their range extension. A similar event occurred in the Tennessee River following impoundment of Kentucky Lake in 1944. None of the three Mississippian species was reported from the Tennessee River by Ortmann (1918, 1924, 1925) or van der Schal- ie (1939) except for Quadrula nodulata, which van der Schalie (1939) listed from Paducah, Kentucky, at the junction of the Tennessee and Ohio rivers. Stansbery (1964) found An- odonta suborbiculata in the Tennessee River at Muscle Shoals, Alabama, in 1963; Isom (1969) found Arcidens confragosus in 1965; and Yokley (1972) reported Quadrula nodu- lata, Arcidens confragosus, and A. suborbicu- lata in Kentucky Lake in 1972. To explain the previous absence of these three species—Anodonta suborbiculata, Arci- dens confragosus, and Quadrula nodulata—in the Cumberland River, one must examine the geological history of the region. Ortmann (1925) pointed out that the Tennessee and Cumberland rivers were at one time joined and separated from the Ohio River, at least for some distance beyond the present confluence. Thornbury (1965) suggested that the two riv- ers had a pre-Pleistocene connection below Paducah, Kentucky, and continued westward through the Metropolis Lowland now occu- pied by the Ohio River, which at that time flowed through the Cache Lowland to the north. The junction with the Ohio River may have occurred near Cairo, Illinois, or perhaps as far south as Memphis, Tennessee. The iso- lation of the Tennessee River from the Cum- berland River by the present Ohio River at the end of the Pleistocene in conjunction with re- duced gradient in the lower sections of the rivers essentially isolated the Cumberlandian fauna in the upper reaches and allowed the invasion of the Ohioan and Mississippian fau- nas to proceed. But, apparently, the direct connection of the Cumberland River to the Ohio River during the Pleistocene was not suf- ficient to allow all of the Ohioan and Missis- sippian species to invade. If length of time alone were the factor limiting the invasion, then all of the common Interior Basin (Mis- sissippian and Ohioan) fauna might have ex- tended into the Cumberland River. Obviously barriers, both ecological and geological, pre- vented wholesale invasion just as they pre- vented most of the Cumberlandian fauna from extending beyond the Tennessee and Cum- berland rivers. Before successful invasion can occur, the appropriate host fish must be pres- ent and a sufficient number of unionids (as larvae, juveniles, or adults) or a gravid female must be transported to a suitable habitat where a reproducing population can become established. Then habitat suitability and lack of competition are required. In fact, the lower Cumberland River may have experienced fluctuations between Ohioan and Cumberlandian species composition since the Pleistocene. During the dry, hypsithermal (Franklin 1994) from 8700 to 5000 years ago, Cumberlandian species may have been more common in the lower Cumberland River, and species such as Epioblasma flexuosa, E. obli- quata, and Pleurobema clava, which may have originated in the Cumberlandian Region, moved into the Ohioan Region. Between 700 and 1000 years ago at least two species, Dro- mus dromas and Epioblasma arcaeformis, were common in the region now the Barkley Dam tailwater (Casey 1987). The changing cli- mate with increasing moisture may have re- stricted the Cumberlandian fauna upstream as Ohioan species invaded the lower reach of the river, which was becoming larger and deeper and more like their natural habitat in the Ohio River. Another notable change from 1911 to the present is the replacement of Quadrula fra- gosa with Q. quadrula. It could be argued that Wilson and Clark (1914) were actually looking at Quadrula quadrula but identified it as Q. fragosa. However, the figures by Coker (1919) clearly distinguish the two and we assume they recognized the difference. We are presently observing a changing species composition from Quadrula quadrula to Q. apiculata (Say, 1829) in Kentucky Lake within the last 15 Unionid Fauna of Lower years, suggesting that these changes can occur rapidly as better-adapted species invade. This, of course, has major implications for attempts at preservation of endangered species. Impoundment and pollution have resulted in a large reduction in the number of native species in the Cumberland River. However, al- tered habitat and reduction in native species have allowed more Interior Basin species, well adapted to impoundment conditions, to invade the lower Cumberland River. There were 30 species reported by Wilson and Clark (1914) in the Cumberland River downstream from CRM 148 that we did not find alive (Table 1). Remnants of these species may survive in the upper regions of Lake Barkley where riverine conditions occur, but it is probably safe to say that they all have dis- appeared from lower sections of the Cumber- land River from approximately Dover, Ten- nessee, CRM 89, to the Ohio River. CONCLUSIONS Twenty-four species of unionids in 18 gen- era still survive in the lower Cumberland Riv- er from mile 47 to mile 2.2. Nine additional species in seven genera were found only as relic shells. No live specimens listed on the federal endangered species list were encoun- tered, although relic or subfossil shells of six endangered species were found: Epioblasma flexuosa, Lampsilis abrupta, Obovaria retusa, Plethobasus cicatricosus, P. cooperianus, and Pleurobema rubrum. Thirty species reported in 1914 were not found alive in our study. It is unlikely that very many of these survive in the Barkley Dam tailwater section of the Cumberland River. Impoundment, both on the Ohio and Cumberland rivers, appears to be the major cause for change in species com- position. However, the changing climate since the Pleistocene, changes in availability of host fish, pollution, and successful invasions by spe- cies from other regions have played various roles in determining the present species com- position. Unionid communities are in constant flux. Better knowledge regarding what con- trols the makeup of unionid communities is needed if their biodiversity is to be preserved. ACKNOWLEDGMENTS Many thanks are extended to Dr. David H. Stansbery, Ohio State University Museum of Cumberland River , Kentucky—Sickel and Chandler 45 Zoology, who assisted in the identification of unionids collected in this project. Many indi- viduals deserve special appreciation for their assistance. The diving team consisted of Garry L. Pharris and the authors with aid from Mark F. Boone, Phil Chandler, and James R. Orr. Commercial shellers assisting with collections were brailers Norris E. Jones, Rudy D. Bridg- es, David Futrell, and Jojo McCloud and di- vers Jim C. Snowden and Daniel R. Thomas. Figure 1 was prepared by Carl Woods. This project could not have been conducted with- out the assistance and financial support of the Nashville District Army Corps of Engineers and the facilities of the Hunter M. Hancock Biological Station of Murray State University. LITERATURE CITED Anderson, R.M., J.B. Layzer, and M.E. Gordon. 1991. Recent catastrophic decline of mussels (Bivalvia: Unionidae) in the Little South Fork Cumberland River, Kentucky. Brimleyana 17:1-8. 1994. Navigation charts Cumberland River Nashville District Smithland, Kentucky to Celina, Ten- nessee. U. S. Army Corps of Engineers, Nashville Dis- trict, Nashville, TN. Bates, J.M. 1962. mussel fauna of Kentucky Reservoir, Tennessee River. Am. Midl. Nat. 68:232-236. Blankenship, S., and D.R. Crockett. 1972. Changes in the freshwater mussel fauna of the Rockcastle River at Liv- ingston, Kentucky. Trans. Ky. Acad. Sci. 33:37-39. Call, S.M., and P.W. Parmalee. 1982. The discovery of extant populations of Alasmidonta atropurpurea (Rat- Anonymous. The impact of impoundment on the inesque) (Bivalvia: Unionidae) in the Cumberland River Basin. Bull. Am. Malacol. Union 1981:42—43. Casey, J.L. 1987. Aboriginal and modern mussel assem- blages of the lower Cumberland River. Southeast. Ar- chaeol. 6(2):115-124. Cicerello, R.R., M.L. Warren Jr., and G.A. Schuster. 1991. A distributional checklist of the freshwater unionids (Bi- valvia: Unionoidea) of Kentucky. Am. Malacol. Bull. 8(2):113-129. Clark, A.H. 1981. The tribe Alasmidontini (Unionidae: Anodontinae), Part I: Pegias, Alasmidonta, and Arci- dens. Smithson. Contrib. Zool. 326. Clark, A-H. 1985. The tribe Alasmidontini (Unionidae: Anodontinae), Part Il: Lasmigona and Simpsonaias. Smithson. Contrib. Zool. 399. Coker, R.E. 1919. Fresh-water mussels and mussel in- dustries of the United States. U.S. Bur. Fish. Doc. 865: 12-89. Franklin, $.B. 1994. Late Pleistocene and Holocene veg- etation history of Land Between the Lakes, Kentucky and Tennessee. Trans. Ky. Acad. Sci. 55:6—-19. Fuller, S.L.H. 1974. Clams and mussels (Mollusca: Bi- 46 Transactions of the Kentucky Academy of Science 57(1) valvia). Pages 215-273 in C.W. Hart Jr. and S.L.H. Ful- ler (eds). Pollution ecology of freshwater invertebrates. Academic Press, New York. Harker, D.F., Jr, M.L. Warren Jr, K.E. Camburn, S.M. Call, G.J. Fallo, and P. Wigley. 1980. Aquatic biota and water quality survey of the upper Cumberland River basin. Kentucky Nature Preserves Commission, Tech- nical Report, Frankfort. Hoeh, W.R. 1990. Phylogenetic relationships among east- erm North American Anodonta (Bivalvia: Unionidae). Malacol. Rev. 23:63-82. Isom, B.G. 1969. The mussel resource of the Tennessee River. Malacologia 7:397-425. Isom, B.G., C. Gooch, and $.D. Dennis. 1979. Redis- covery of a presumed extinct river mussel, Dysnomia sulcata (Unionidae). Nautilus 93(2—3):84. Johnson, R.I. 1978. Systematics and zoogeography of Pla- giola (=Dysnomia = Epioblasma), an almost extinct ge- nus of freshwater mussels (Bivalvia: Unionidae) from middle North America. Bull. Mus. Comp. Zool. Harv. Univ. 148(6):239-320. Johnson, R.I. 1980. Zoogeography of North American Unionacea (Mollusca: Bivalvia) north of the maximum Pleistocene glaciation. Bull. Mus. Comp. Zool. Harv. Univ. 149(2):77-189. Miller, A.C., L. Rhodes, and R. Tippit. 1984. Changes in the naiad fauna of the Cumberland River below Lake Cumberland in central Kentucky. Nautilus 98:107—110. Neel, J.K., and WR. Allen. 1964. The mussel fauna of the upper Cumberland Basin before its impoundment. Malacologia 1(3):427-459. Ortmann, A.E. 1918. The Nayades (freshwater mussels) of the upper Tennessee drainage with notes on syn- onymy and distribution. Proc. Am. Philos. Soc. 57(6): 521-625. Ortmann, A.E. 1924. The naiad-fauna of Duck River in Tennessee. Am. Midl. Nat. 9:18-62. Ortmann, A.E. 1925. The naiad-fauna of the Tennessee River system below Walden Gorge. Am. Mid]. Nat. 9: S21=372: Ortmann, A.E., and B. Walker. 1922. On the nomencla- ture of certain North American Naiades. Occas. Pap. Mus. Zool. Univ. Mich. 112:1—75. Parmalee, P.W., W.E. Klippel, and A.E. Bogan. 1980. Notes on the prehistoric and present status of the naiad fauna of the middle Cumberland River, Smith County, Tennessee. Nautilus 94(3):93-105. Schmidt, J.E. 1982. The freshwater mussels of the Stones River above J. Percy Priest Reservoir, Tennessee. U.S. Army Corps of Engineers, Nashville District, Nashville, TN. Schuster, G.A., R.S. Butler, and D.H. Stansbery. 1989. A survey of the unionids (Bivalvia: Unionidae) of Buck Creek, Pulaski County, Kentucky. Trans. Ky. Acad. Sci. 50:79-85. Stansbery, D.H. 1964. The Mussel (Muscle) Shoals of the Tennessee River revisited. Annu. Rep. Am. Malacol. Union 1964:25-28. Stansbery, D.H. 1970. Eastern freshwater mollusks (1), the Mississippi and St. Lawrence river systems. Proc. Amer. Malacological Union Symposium on Rare and Endangered Mollusks. Malacologia 10(1):9-22. Starnes, L.B., and A.E. Bogan. 1982. Unionid mollusca (Bivalvia) from Little South Fork Cumberland River, with ecological and nomenclatural notes. Brimleyana 8: 101-119. Starnes, L.B., and A.E. Bogan. 1988. The mussels (Mol- lusca: Bivalvia: Unionidae) of Tennessee. Am. Malacol. Bull. 6(1):19-37. Thornbury, W.D. 1965. Regional geomorphology of the United States. John Wiley and Sons, New York. Turgeon, D.D., A.E. Bogan, E.V. Coan, W.K. Emerson, W.G. Lyons, W.L. Pratt, C.F.E. Roper, A. Scheltema, F.G. Thompson, and J.D. Williams. 1988. Common and scientific names of aquatic invertebrates from the United States and Canada: mollusks. Am. Fish. Soc. Spec. Publ. 16. [USFWS] U.S. Fish and Wildlife Service. 1993. [Endan- gered and threatened wildlife and plants.] [U.S.] Code of Federal Regulations 50 Ch. 1 (10-1-94 Edition) §17.11, §17.12. van der Schalie, H. 1939. Additional notes on the Naia- des (freshwater mussels) of the lower Tennessee River. Am. Midl. Nat. 22(2):452-457. Wilson, C.B., and H.W. Clark. 1914. The mussels of the Cumberland River and its tributaries. U.S. Bur. Fish. Doc. 781. Wilson, E.O. 1994. Biodiversity: challenge, science, op- portunity. Am. Zool. 34:5-11. Yokley, P., Jr. 1972. Freshwater mussel ecology, Kentucky Lake, Tennessee. May 1, 1969-June 15, 1972. Tennes- see Game and Fish Commission. Project 4-46-R. Nash- ville, TN. Trans. Ky. Acad. Sci. 57(1):47-48. 1996. NOTES Nesting Association of the Cyprinid Fishes Phoxi- nus cumberlandensis and Semotilus atromaculatus (Cyprinidae).—Phoxinus cumberlandensis, a cyprinid en- demic to small, upland streams in the upper Cumberland River basin of eastern Kentucky and Tennessee (1), is known to spawn only over fine, silt-free gravel in Cam- postoma anomalum nests (2). In silt-free streams, P. cum- berlandensis presumably spawns over gravel in riffles or runs. However, siltation and habitat loss from coal mining, silviculture, agriculture, and road construction have re- duced and fragmented P. cumberlandensis habitat and re- sulted in the fish being a threatened species by the U.S. Fish and Wildlife Service (3). Herein we report observa- tions of an additional host species for P. cumberlandensis. On 12 May 1993 we observed and photographed ca. 30 brilliantly colored P. cumberlandensis in an occupied Se- motilus atromaculatus nest in Rock Creek, McCreary County, Kentucky. Rock Creek is a second-order tributary to Jellico Creek; it supports one of the best remaining populations of P. cumberlandensis in Kentucky (Cicerello and Laudermilk, unpubl. data). Upstream from the site of our observations, the watershed is densely forested and contains only one small, reclaimed strip mine along the Tennessee border. As a result, the sand, gravel, and scat- tered cobbles underlying the stream’s alternating pools and short riffles are relatively silt-free. At the time of our observations, Rock Creek was clear, low, and ca. 3-4 m wide; flow was slow to negligible. The gravel Semotilus nest was at a depth of ca. 7-10 cm in the lower end of a pool immediately upstream from a small riffle. When we approached the stream, the nest’s occupants fled to the deeper, upper end of the pool, but the P. cum- berlandensis aggregation slowly meandered back toward the nest after we took cover. Members of the aggregation included brilliantly colored males and females and smaller, apparently juvenile individuals lacking bright spawning colors. While moving toward the nest, males chased fe- males individually and in groups in the manner described by Starnes and Starnes (2). After an estimated 10-15 min- utes, the aggregation returned to and hovered over the ca. 25 cm diameter nest depression. About 5-10 minutes lat- er, a nuptial male Semotilus (ca. 18 cm TL) returned to the nest where his presence was obscured by the P. cum- berlandensis hovering above. Starnes and Starnes (2) wit- nessed aggressive behavior toward Semotilus by nesting C. anomalum, which they believed provided associated P. cumberlandensis with some protection from egg preda- tion. We did not observe spawning by P. cumberlandensis, but their behavior and previously unreported association with a nesting S. atromaculatus (4) are consistent with spawning by P. cumberlandensis observed in a C. anom- alum nest (2) and by P. oreas in Nocomis nests (5). Phoxinus cumberlandensis occurs syntopically with Se- motilus and C. anomalum above Cumberland Falls, and with Semotilus and C. oligolepis, a probable P. cumberlan- densis nesting host, below the falls. However, Semotilus 47 occurs more frequently and is more abundant than Cam- postoma spp. in streams inhabited by P. cumberlandensis. Of 95 collections of P. cumberlandensis made in 1993- 1994, 89 (94%) included Semotilus but only 24 (25%) in- cluded Campostoma spp. During April through June, when P. cumberlandensis spawns (2), Semotilus also was present more often than Campostoma spp. (51 collections Way, ILID)). We believe these observations strongly suggest that P. cumberlandensis spawns in Semotilus atromaculatus nests, even in relatively silt-free streams such as Rock Creek; that Semotilus is more important than Campostoma spp- in providing spawning habitat for P. cumberlandensis; and that nest-building cyprinids probably play an important role in conservation of P. cumberlandensis by providing spawning habitat in relatively clean streams as well as those degraded by silt. However, additional observations are needed to confirm these findings and to document spawning by P. cumberlandensis independently of nest- building cyprinids. We thank B.M. Burr (Southern Illinois University at Carbondale) for reviewing the draft, and V. Bishop and L. Perry (U.S. Forest Service) for providing land-use infor- mation. This study was supported in part by the Kentucky Department for Surface Mining Reclamation and En- forcement, Frankfort, Kentucky. LITERATURE CITED. (1) Starnes, W. C., and L. B. Starmes. 1978. A new cyprinid of the genus Phoxinus en- demic to the upper Cumberland River drainage. Copeia 1978:508—516. (2) Starnes, L. B., and W. C. Starnes. 1981. Biology of the blackside dace Phoxinus cumberlandensis. Am. Midl. Nat. 106(2):360-371. (3) U.S. Fish and Wildlife Service. 1987. Endangered and threatened wildlife and plants; determination of threatened species status for the blackside dace. Fed. Reg. 52:22580-22585. (4) Johnston, C. E., and L. M. Page. 1992. The evolution of complex reproductive strategies in North American minnows (Cy- prinidae). Pages 600-621 in R. L. Mayden (ed). System- atics, historical ecology, and North American freshwater fishes. Stanford Univ. Press, Stanford, CA. (5) Raney, E. C. 1947. Nocomis nests used by other breeding cyprinid fishes in Virginia. Zoologica 32:125-132—Ronald R. Ci- cerello and Ellis L. Laudermilk, Kentucky State Nature Preserves Commission, 801 Schenkel Lane, Frankfort, KY 40601. Sclerochloa dura (Poaceae) in Kentucky.—The Eu- ropean grass Sclerochloa dura (L.) Beauv. (Figure 1), hard grass, was first collected in North America in New York in 1895, but this introduction apparently did not lead to naturalization of the species. Thirty-three years later, in 1928 in Utah, the grass was collected again. Since then, 48 Transactions of the Kentucky Academy of Science 57(1) Figure 1. Sclerochloa dura. A, plant, < 0.45. B, spikelet, x 5.0. C, caryopsis, X 7.2. Drawing by Paul W. Nelson. the plant has become fully naturalized in the United States, having been found in 16 additional states from coast to coast by 1991 (1). The species has since been recorded from Illinois and Indiana (2) and from Iowa (3). It can now be added to the flora of Kentucky, where it is known from six counties (Bracken, Campbell, Grant, Jef- ferson, Kenton, Robertson), all in the northern part of the state. The specimens were collected from mid-april to mid-May. All the Kentucky specimens were growing in athletic fields and associated disturbed areas. Of the 12 such fields searched in 1995, the grass was in nine of them. In some places there were hundreds of individuals; in others, in- dividuals were few and widely scattered. The recent—and first—record from Ohio (Hamilton County) (1) was also from an athletic field. Swink and Wilhelm (2) mentioned that S. dura is common in fairgrounds in parts of its range. The first lowa collection was from a gravel parking lot (3). We suggest that S$. dura may be carried from field to field in soil on the shoes of ball players as the players peregrinate in search of victories. The grass turns brown and dies early in the season, providing abundant propa- gules in places frequented by players and onlookers: the fields, the bleacher areas, and the nearby parking lots. We also suggest that the species might also be spread in “athletic field” turfgrass mixtures. Many of these mixtures are said to originate in Washington and Oregon, where S. dura has been known for about 60 years. We are unable to confirm or deny this idea. Associates of the plant in Kentucky include other vernal annuals: Capsella bursa-pastoris, Cerastium spp., Draba brachycarpa, D. verna, Holostewm umbellatum, Matricar- ia matricarioides, Poa annua, P. chapmaniana, Stellaria media, Veronica peregrina, and V. arvensis. The grass often grows with Poa annua, which bears a resemblance to it, especially in vegetative condition. Both grasses come into flower when they are quite young; they are then easily distinguished by their inflorescences, which in hard grass are distinctly one-sided, quite unlike the more or less symmetric ones of P. annua. The branches of S. dura are ascending to prostrate, forming clumps to 12 cm wide. We cite here a voucher specimens for the presence of S. dura in Kentucky: KENTUCKY. Jefferson Co., Louis- ville, in “turf” of Poa annua in mowed ballfield along River Road near Ohio Street, 26 Apr 1994, Thieret 57202 (KNK). LITERATURE CITED. (1) Brandenburg, D. M., J. R. Estes, and J. W. Thieret. 1991. Hard grass (Sclerochloa dura, Poaceae) in the United States. Sida 14:369-376. (2) Swink, F., and G. Wilhelm. 1994. Plants of the Chicago region, 4th ed. Indiana Academy of Science, Indianapolis. (3) Wilson, B. L. 1992. Checklist of the vascular flora of Page County, Iowa. J. Iowa Acad. Sci. 99:23-33.—David M. Brandenburg, The Dawes Arboretum, 7770 Jackson- town Road, S.E., Newark, OH 43055; John W. Thieret, Department of Biological Sciences, Northern Kentucky University, Highland Heights, KY 41099. Trans. Ky. Acad. Sci. 57(1):49-66. 1996. Abstracts of Some Papers Presented at the 1995 Annual Meeting of the Kentucky Academy of Science AGRICULTURAL SCIENCES Contents of field-grown turnips in relation to colored mulches. G. S. ANTONIOUS* and M. E. BYERS, At- wood Research Facility, Kentucky State University, Frank- fort, KY 40601; M. J. KASPERBAUER, Coastal Plains Research Center, USDA-ARS, Florence, SC 29501. Growth, development, and chemical composition of field-grown turnips (Brassica campestris cv. Purple Top) can be influenced by the spectrum of light reflected from the soil surface. The shoot/root weight ratio and the chem- ical composition of roots were affected by the quantity of blue light and the ratio of far-red relative to red (FR/R) in upwardly reflected light. A FR/R ratio higher than the ratio in incoming light generally resulted in a greater shoot/root biomass ratio. In a taste test of uncooked roots, 24 of 25 panelists indicated that turnips that received more reflected blue light during development had roots with a sharper flavor. The majority of the tasters also in- dicated that roots from plants grown with green mulch had the mildest (almost sweet) flavor. In a 2-year labora- tory study, roots from turnips grown in field plots over blue, green, silver, and white surfaced plastic mulches were analyzed for concentrations of total glucosinolates (GSLs), ascorbic acid, and sugar. The greatest concentra- tions of GSLs and ascorbic acid were found in roots grown with blue mulches. Reducing sugars were higher in roots grown with green than in those grown with blue mulches. The comparison of chemical composition of roots from plants grown with blue versus green mulches is important because both blue and green surfaces reflected about the same amount of photosynthetic light, very little red light, different amounts of blue, and about the same FR/R ratio. The quantity of blue in reflected light received by leaves contributed to chemical differences related to flavor of roots from turnip plants grown in sunlight over blue- ver- sus green-surfaced mulches. Differences in chemical com- position of roots grown with white or silver mulches (both of which reflected more photosynthetic light and lower FR/R ratios than the blue or green mulches) were less pronounced, and concentrations were generally lower than those in roots grown with blue mulch. Dermal and inhalation exposure of several mixers/ap- plicators to ag-chemicals during tobacco production. J. A. LONSWAY,* M. E. BYERS, H. A. DOWLA, and M. PA- NEMANGALORE, Community Research Service, Ken- tucky State University, Frankfort, KY 40601. Exposure of limited-resource tobacco-farmers to ag- chemicals and natural products has been identified as an area for concern. This study was designed to determine routes and extent of exposure to insecticides and growth regulators that mixers/applicators would encounter using 49 a tractor-mounted boom-sprayer or a highboy. Dermal and inhalation exposures were determined using a gauze pad monitoring technique and personnel type air samplers with charcoal adsorbents, respectively. Endosulfan, ace- phate, and its metabolite methamidophos were analyzed by gas liquid chromatography (GLC) and GLC/mass spec- trometry (MS). Results from 1994 field studies indicated that dermal exposure to acephate was greater for mixing than for spraying. For spraying, dermal exposure to meth- amidophos well exceeded exposure to acephate. This dif- ference was likely due to acephate chemically transform- ing into methamidophos during spraying. Inhalation val- ues ranged from 0 to 3.9 mg/kg/hr for acephate and meth- amidophos. Some exposure to nicotine was also assessed. Field experiments were repeated in 1995. Effects of waste composts on tomato production in con- tainers. BRIAN D. LACEFIELD* and ELMER GRAY, Department of Agriculture, Western Kentucky University, Bowling Green, KY 42101. The increasing waste production by municipalities and the growing interest in gardening by urbanites amply jus- tify research on compost utilization in container garden- ing. Tomatoes are well suited for container production. Objectives of the present study were to compare two to- mato cultivars—Patio and Celebrity—for production in containers and to compare different waste composts— leaf, brush, and N-Viro-Soil—for their effects on tomato yield. Ten growth media consisted of soil and the three composts alone plus 50:50 (by volume) mixtures of all combinations of the four media. Containers were plastic barrel sections (58 cm diameter, 38 cm depth) with ca- pacity of 64 liters. Forty containers permitted four repli- cations of the 10 growth media. Media fertilization con- sisted of one pre-transplant application, at kg/ha rates, of 56.0 N, 24.5 P, 46.5 K. Vine-ripe tomatoes were harvested twice weekly during the production season. Celebrity pro- duced significantly fewer and significantly heavier fruits than Patio, resulting in no significant difference between cultivars for total season production per plant. There were no significant differences in tomato yields associated with growth media. There also were no significant cultivar X growth media interactions. These preliminary results in- dicated that composts of waste products can be effectively substituted for soil in container production of tomatoes. Pawpaw research at Kentucky State University: an up- date. DESMOND R. LAYNE, Atwood Research Facility, Kentucky State University, Frankfort, KY 40601. Kentucky State University (KSU) currently has a full- time research program directed at developing pawpaw (Asimina triloba) as a new commercial fruit crop for Ken- 50 Transactions of the Kentucky Academy of Science 57(1) tucky and the United States. Three concurrent research projects are underway at KSU: (1) overcoming the horti- cultural limitations to development as a new crop; (2) characterizing the morphological and molecular variation in a diverse germplasm collection; and (3) developing a centralized research support system. For project 1, we have conducted greenhouse experiments to establish op- timal conditions (light regimen, fertilization, soil temper- ature, etc.) for production of robust seedlings. A new paw- paw regional variety trial orchard (300 trees) comparing the most promising 28 pawpaw clones or named varieties was also planted. This replicated research trial involves cooperators in 15 states who will plant identical orchards and evaluate clone suitability, etc. for their region. For project 2, we have planted a new orchard (1200 trees) of over 400 pawpaw accessions from 16 states. In addition, several inter- and intraspecific hybrids were planted in this same orchard for long-term evaluation. Molecular evalu- ation of isozyme polymorphisms began fall 1995. Project 3 began fall 1995 and is twofold in nature. First, we will develop an on-line computer database for information on any subject related to pawpaws. Second, we will develop improved techniques for seed propagation and clonal (vegetative) propagation. As a result of federal funding of the latter project, KSU has been designated the National Clonal Germplasm Repository for pawpaws. Phenotypic variation in Indian mock strawberry (Du- chesnea indica). ELMER GRAY* and BRIAN D. LACE- FIELD, Department of Agriculture, Western Kentucky University, Bowling Green, KY 42101. Indian mock strawberry, Duchesnea indica, is common on uncultivated open land in central and eastern U.S. Un- der conditions resulting from relaxed lawn management, the plant may dominate areas of lawns. It reproduces pro- lifically through vegetative propagules and seeds. The rel- ative proportions of asexual and sexual reproduction im- pact variation in the resulting population. The objective of the study was to characterize phenotypic variability of local and more distant populations of the plant. Measur- able plant traits included petiole length, leaflet length and width, and leaflet serrations. Linear correlations (r) among these traits were highly significant in each of the nine pop- ulations. However, relationships among the traits were not the same in the different populations as evidenced by sig- nificant heterogeneity among the correlation coefficients and variability among the coefficients of variation (stan- dard deviation < 100/mean). Overall, the results indicated a high level of phenotypic plasticity in Indian mock straw- berry resulting from both genetic and environmental ef- fects. Testing alfalfa varieties for yield. LINDA G. BROWN,* Department of Agriculture, Western Kentucky University, Bowling Green, KY 42101; J. C. HENNING and L. M. LAURIAULT, Department of Agronomy, University of Kentucky, Lexington, KY 40506. The Kentucky Alfalfa Variety Trials are a continuing co- operative project between Western Kentucky University and the University of Kentucky. Alfalfa is an important crop in cash hay production enterprises of this state. Every year dozens of new varieties are released to the public for sale, making the task of selecting an appropriate variety for planting very challenging for producers. Each variety is distinctly different from others in one of the following ways: maturity date, dormancy rating, growth habit, num- ber of leaflets per leaf, pest resistance, and/or yield po- tential. The objective of this research is to evaluate the yield performance of new alfalfa varieties at several loca- tions in Kentucky. Plots are established at four locations in Kentucky on a rotating basis: Bowling Green, Lexing- ton, Princeton, and Mayslick. Cumulative yield data will be presented as well as the recommended procedure for using the data to select a variety. Tobacco field worker re-entry exposure to acephate, nicotine, and their metabolites. N. C. BAKER,* J. A. LONSWAY, M. E. BYERS, H. A. DOWLA, and M. PA- NEMANGALORE, Community Research Service, Ken- tucky State University, Frankfort, KY 40601. Field workers were monitored for dermal and respira- tory exposure to acephate and its metabolite methami- dophos at re-entry intervals of 2, 4, 8, 24, and 48 hours after application. The task of field workers was to “top” bloom stage tobacco. Exposure to nicotine was also as- sessed. Dermal exposure was measured by analyzing 10 gauze pads attached to the clothing of workers to repre- sent human body regions. Hand exposure was determined using cotton gloves. Respiratory exposure was determined using personnel type air samplers equipped with charcoal adsorbent traps. Gas liquid chromatography was used to quantify residues of acephate, methamidophos, and nico- tine in pads, gloves, and charcoal air sampler adsorbents. Dermal exposure was greatest for the hands at all re-entry intervals. There was not a consistent trend of decreasing pesticide residue over time. This can probably be attrib- uted to heavy dew or rainfall on the tobacco plants for 24 and 48 hour re-entries. Nicotine exposure was random by body region, but residues increased consistently with moisture levels. Our preliminary results suggest that a 48- hour post-spraying waiting period may not be sufficient. BIOCHEMISTRY & PHARMACOLOGY Effect of manipulation of central histaminergic activity on locomotor activity in rats). DANITA SAXON KEL- LEY,* Department of Consumer and Family Sciences, Western Kentucky University, Bowling Green, KY 42101; L. PRESTON MERCER, Department of Nutrition and Food Science, University of Kentucky, Lexington, KY 40506. Physical activity is reported to have a circadian rhythm (period of about 24 hours) in rats and humans. This study examined the relationships between locomotor activity, diet composition, and histaminergic activity in the central nervous system (CNS). Dietary treatments altering the histaminergic parameters of central histamine and/or H,- Abstracts, 1995 Annual Meeting 51 receptor concentrations were performed. Male rats were ad libitum fed a 25% casein diet (NP), ad libitum fed a 1% casein diet (LP), or pair-fed a 25% casein diet (PF). Significant ultradian rhythms (period < 20 hours), in ad- dition to diurnal changes, in locomotor activity were iden- tified for all treatment groups. Pair-fed rats had increased, though not significant, maximum activity compared to NP rats. Administration of the H, antagonist doxepin signifi- cantly decreased mean activities of LP and PF rats and significantly decreased maximum activity of PF rats. This study supports the involvement of CNS histaminergic ac- tivity in locomotor activity at the H,-receptor. Effects of chloropropanes on glycolysis. YI ZHOU and ROBERT F. VOLP,* Department of Chemistry, Murray State University, Murray, KY 42071. Chloropropanes are a group of organic chemicals pro- duced in large quantities for a variety of uses. Human exposure occurs during the production and use of these compounds and via contaminated groundwater. The tox- icity of chloropropanes has received little attention al- though analogies to chloroethanes suggest that chloropro- pane toxicity is determined by their biotransformation. Structural similarity between putative chloropropane bio- transformation products and intermediates of glycolysis suggests that toxicity occurs when chloropropane biotrans- formation products inhibit glycolytic enzymes. To inves- tigate the interactions of chloropropanes, their biotrans- formation products, and glycolytic enzymes, rat liver cy- tosol was incubated at pH 7.5 and 37°C with 3-phospho- glycerate (3PG) and necessary cofactors. Glycolytic reactions initiated by 3PG were monitored by measuring NADH absorbance at 340 nm. The effects of 1,2,3-tri- chloropropane (TCP; 0.5, 5.0 mM), glutathione (GSH; 1.0 mM), and rat liver microsomes (0.1, 0.5 mg/ml) were test- ed by adding each individually and in all possible combi- nations. In the absence of microsomal protein, TCP did not affect glycolysis rate, suggesting that biotransforma- tion is necessary for an effect. With 0.1 mg/ml microsomal protein, TCP decreased glycolysis rate; inclusion of GSH abolished this effect. These results suggest that a micro- somal biotransformation product of TCP inhibits glycoly- sis and that GSH conjugation detoxifies either this product or TCP itself. With 0.5 mg/ml microsomal protein, TCP did not affect glycolysis rate in the absence of GSH; it increased the rate in the presence of GSH. An explanation for these effects is likely to require identification of mul- tiple effects mediated by the microsomes. BOTANY & MICROBIOLOGY Analysis of morphological and genetic variation in Hex- astylis contracta. PATRICK E. CARROLL* and ZACK E. MURRELL, Department of Biology, Western Ken- tucky University, Bowling Green, KY 42101. Hexastylis contracta, an herbaceous evergreen plant in the Aristolochiaceae, is generally restricted to remnant old-growth hemlock forests in the Cumberland Plateau of Kentucky and Tennessee, with three disjunct populations in western North Carolina. Extensive field work identified 18 new populations of H. contracta in Tennessee. The apparent hybrid found appears to be the result of crosses between H. contracta and H. arifolia. Genetic variability within H. contracta and the putative hybridization be- tween H. contracta and H. arifolia are the subjects of on- going molecular analyses of the internal transcribed spacer (ITS) regions of nuclear ribosomal DNA. The putative hy- brid exhibits floral and leaf similarities with H. arifolia, but the fleshy perianth has a flare at or below the middle that is very similar to the H. contracta perianth structure. The hypothesis of hybridization is explored through mor- phometric analyses of leaf and perianth structure of the putative hybrid and the parent species. Blue- and white-fruited dogwoods (Cornus): internal transcribed spacer (ITS) region sequence data and species relationships. JEFFREY J. BAKER* and ZACK E. MUR- RELL, Department of Biology, Western Kentucky Uni- versity, Bowling Green, KY 42101. The genus Cornus is currently recognized with eight segregate subgenera and ca. 50 species. The blue- and white-fruited dogwoods include about 30 species that have traditionally been segregated, on the basis of leaf position and branching patterns, into opposite-leaved (subg. Kran- iopsis) and alternate-leaved (subg. Mesomora) groups. Re- cent studies have separated the Asian blue-fruited species C. oblonga and the South American blue-fruited species C. peruviana into segregate subgenera or sections. Our study involves analyses of relationships in Cornus using the internal transcribed spacer (ITS) region of nuclear ri- bosomal DNA. The sequences were generated using cycle sequencing on an automated sequencer, aligned using CLUSTAL, and analyzed using PAUP. The alignments, with the exception of length variation in C. oblonga and C. peruviana, showed a high degree of similarity. The trees produced by the PAUP analyses suggest that the blue-fruited dogwoods are monophyletic, with the excep- tions of C. oblonga and C. peruviana, which appear to be basal lineages within Cornus. Within the monophyletic clade of blue- and white-fruited dogwoods, there is strong support for an alternate-leaved clade and an opposite- leaved clade. The opposite-leaved clade can be divided into two monophyletic groups, the “stolonifera group” and the “stricta group.” The support for these groups based upon molecular data is in agreement with morphological data from plant architecture and leaf venation patterns. Dogwood (Cornus) subgeneric relationships: evidence from the internal transcribed spacer (ITS) of nuclear ri- bosomal DNA. ZACK E. MURRELL* and JEFFREY J. BAKER, Department of Biology, Western Kentucky Uni- versity, Bowling Green, KY 42101. Molecular sequence data from the internal transcribed spacer (ITS) region of 18-26S nuclear ribosomal DNA have been derived for eight subgenera of the genus Cor- nus and from representatives of the putative outgroups Nyssa, Alangium, and Davidia. These sequences show ex- 52 Transactions of the Kentucky Academy of Science 57(1) treme variations in length, from 250 to 350 bases in both ITS I and ITS II. Attempts have been made to align these data using the computer program CLUSTAL, as well as manual alignment; results suggest that multiple repeats and length variation are difficult to align using CLUSTAL. The manually aligned complete data set was analyzed us- ing PAUP, and subsets of this data set, omitting the IN- DELS, were also analyzed and compared to the total data set. The results of these analyses support the monophyly of the blue-fruited dogwoods and also the sister relation- ship of the big-bracted dogwoods and the cornelian cher- ries. Evolutionary positions of C. oblonga, C. peruviana, and C. volkensii in relationship to the remainder of Cor- nus are equivocal, largely due to the extreme length vari- ation of their sequences and the effects of long branch attraction. Molecular analysis of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA in a dogwood (Cornus) hybrid complex. CHRISTOPHER P. REED* and ZACK E. MURRELL, Department of Biology, West- em Kentucky University, Bowling Green, KY 42101. The distributions of two species of blue-fruited dog- woods, Cornus amomum and C. obliqua, have a broad area of sympatry, from Maine to Tennessee. Within this area, putative hybrids have been identified on the basis of several morphological characters. Although the habit, per- iderm, and inflorescences are similar in the two, the leaf pubescence and abaxial epidermal protuberances can be used to separate them; these characters are intermediate in the putative hybrids. Molecular sequence data derived from the internal transcribed spacer region (ITS) of 18— 26S nuclear ribosomal DNA has been generated for the putative hybrids using Polymerase Chain Reaction (PCR) and standard dideoxy sequencing. Initial sequence data exhibit double banding at several nucleotide sites, sug- gesting a lack of uniformity among a putative hybrid’s mul- tiple ITS copies. Ongoing cloning studies of amplified PCR products from the putative hybrids are being se- quenced in exploration of this multiple copy region. This aspect of the research is an attempt to test the use of cloned PCR amplified products as a method to detect hy- brids and hybridization. The generated data are being used to determine the extent of hybridization between the two species. Preliminary bryoflora of an old growth forest, Pine Mountain, Kentucky. JUDITH E. ROZEMAN,* Office of Institutional Research and Effectiveness, Berea College, Berea, KY 40404; TIMOTHY J. WECKMAN, Depart- ment of Biological Sciences, Eastern Kentucky University, Richmond, KY 40475. The bryophytes, including the mosses, liverworts, and hornworts, have been undercollected compared to vas- cular plants in Kentucky. Fewer than 2,500 vouchered specimens are known to be on deposit in Kentucky her- baria, representing about one percent of total plant col- lections. A recent checklist of Kentucky bryophytes in- cludes nearly 300 taxa of mosses and 105 hepatics and hornworts. The primary purpose of the present study is to provide a preliminary bryoflora of the Blanton Forest, a 2,300-acre tract of old growth woods located on the south face of Pine Mountain near Harlan, Kentucky. The secondary purpose of the project is to augment the num- ber of vouchered bryophyte specimens in the state. Col- lections have been made in six major habitats including bogs and seeps, high elevation xeric sites, mixed meso- phytic communities, hemlock-beech communities, creeks, and rock houses and rock outcrops in both wet and dry conditions. Specific substrates include rock, soil, water systems, decaying trees and nursery logs, and living trees. Collections have been made over a 2-year period in all seasons. All phases of the work (collections and identifi- cations) are still in progress. Preliminary results have yielded 58 genera of mosses (the liverwort collection is still to be examined). The degree of similarity of the gen- era list with that of Lilley Cornett Woods, a comparable tract of old growth forest in eastern Kentucky, is 96%. The degree of similarity at the species level is still to be de- termined. Notable finds include Andreaea rothii, Cam- pylopus tallulensis, Dicranodontium denudatum, and Fon- tinalis sullivantii. Seed dispersal in Rhus aromatica. XIAOJIE LI,* JER- RY M. BASKIN, and CAROL C. BASKIN, School of Bi- ological Sciences, University of Kentucky, Lexington, KY 40506. Attempts were made to quantify dispersal of the 1-seed- ed drupaceous fruit in Rhus aromatica. In 1994, obser- vations began on 12 June. By 3 July, 48.3 + 7.0% (« + SE) of the fruits had been dispersed; the percentage reached 99.9 + 0.1 by 13 August. Compared to 1994, rate of dispersal began faster but finished slower in 1995. On 19 June, 51.5 + 6.1% of the fruits had been dispersed; by 3 July, the percentage was 87.2 + 2.8. About 5.5% of the fruits were still on the mother plants on 4 September. During the dispersal period, fruits and/or seeds (i.e., seed plus endocarp) were collected in seed traps on the ground, mostly at weekly intervals. The fruit/seed rain was 66.3 m2 in 1994 and 62.5 m2 in 1995, with the seed com- ponent being 77.8 + 8.3% and 79.7 + 8.6%, respectively. The percentages of fruits and seeds that disappeared from the seed traps due to predation/emigration were 84.2 + 7.3 and 71.7 + 8.0, respectively, leaving a transient seed bank of 18.2 seeds m~ in 1994 and 19.1 seeds m~? in 1995. An additional loss of seeds from the transient seed bank was due to natural softening of the endocarp. After 1 year of burial in the field, 29.4 + 2.7% of the seeds had softened. Thus, the input to the persistent seed bank was 12.8 seeds m~? in 1994 and 13.5 seeds m~? in 1995. Species delineation and population structure of Spiraea virginiana. JAMES C. ESTILL* and ZACK E. MUR- RELL, Department of Biology, Western Kentucky Uni- versity, Bowling Green, KY 42101. Spiraea virginiana is a rare rhizomatous shrub found in Abstracts, 1995 Annual Meeting 53 heavily scoured areas of high-gradient streams in the southern Blue Ridge and Appalachian Plateau provinces and the edge of the Blue Grass Province in Kentucky. Although the species is distributed in six states, the ma- jority of known individuals occur in Kentucky, Tennessee, and West Virginia. An intensive field search for S. virgi- niana in Tennessee has yielded 22 new populations to add to the 13 previously known Tennessee populations. Sev- enteen new populations were located in four drainages that contained known populations, and five populations were found in three “new” drainages. Three known pop- ulations, not relocated, are presumed extirpated. All other known populations were relocated and appeared relatively unchanged in number and density since they were last surveyed, Examination of molecular and morphological character variation suggests that the three species of sec- tion Calospira found in southeastern North America—S. corymbosa, S. japonica, and S. virginiana—can be rec- ognized as segregate species. Previous germination stud- ies, examination of total seed production, and field anal- yses of populations suggest that S. virginiana is limited to asexual reproduction. Ongoing studies utilizing molecular markers address the issues of population and drainage ge- netic identity of the species. Survey of Ohio River bacteria: Gram-negative organ- isms. STEVE KRISTOFF, Department of Biology, Tho- mas More College, Crestview Hills, KY 41017. The Ohio River is extensively used for transportation, recreation, and drinking and industrial water. Although co- liform studies of the river are performed on a regular ba- sis, a literature search reveals little information concerning the bacterial flora. As knowledge of the role of microor- ganisms in the environment grows, it becomes clear that knowledge of the bacterial flora of the river can contribute to our understanding of the river as an ecosystem. Also, since thousands of people use the river for recreational purposes (including physical contact with the water) and as a source of drinking water, it is clear that the river has a significant impact on public health. This survey was un- dertaken with these concerns in mind. Samples were taken from the Ohio River at the Thomas More College Biology Field Station in Campbell County, Kentucky (Riv- er Mile 451). Our preliminary data indicate that the ma- jority of cultivable Gram-negative organisms are Pseudo- monas, with smaller numbers of Acinetobacter, and oc- casional members of the Enterobacteria group. A test of susceptibility to antimicrobial agents reveals that a major- ity of the Pseudomonas are resistant to several classical agents including ampicillin, streptomycin, and chloram- phenicol. Most, however, are susceptible to tetracycline. Symbiotic nitrogen fixation by the turfgrass weed black medic (Medicago lupulina). DAVID LOWELL ROBIN- SON,* Biology Department, Bellarmine College, Louis- ville, KY 40205; CARROLL P. VANCE, Agronomy and Plant Genetics Department, and USDA-ARS, Plant Sci- ences Research Unit, University of Minnesota, St. Paul, MN 55108. Black medic (Medicago lupulina), a common weed of commercial and residential turfgrass in the Midwest, is a close relative of alfalfa (Medicago sativa), a forage crop known for its high rates of atmospheric nitrogen (N) fix- ation. This process is due to a symbiotic relationship be- tween plant and soil bacteria (Rhizobium meliloti) that re- sults in formation of N-fixing root nodules. Previous re- search has shown that, in alfalfa, N fixation rates decline when top growth is frequently harvested, when N fertil- izers are applied to soil, or when the plant blooms. All three of these conditions occur in turf where black medic is a weed. Therefore, we evaluated nodulation and N fix- ation in black medic after frequent harvesting, N appli- cations, and flowering to evaluate its N-fixing potential un- der these conditions. Although frequent harvesting of the shoot caused N fixation rates to decline, black medic nod- ules still fixed significant amounts. Nitrate applications to black medic and alfalfa caused a greater decline in no- dulation of the latter than the former. Therefore, it ap- pears that black medic maintains the potential to fix N under common turf management scenarios. This infor- mation is useful not only in describing the ecological role of this weed in turf but adds to our understanding of sym- biotic N fixation. CHEMISTRY Evaluation of the Toledo Chemistry Placement Exam— Form 1981. L. C. BYRD and D. R. HARTMAN,* De- partment of Chemistry, Western Kentucky University, Bowling Green, KY 42101. Attempts to identify under-prepared students encour- aged investigations into correlation of success in college chemistry with high school grade-point average, class rank or chemistry grade, ACT, SAT-math, aptitude test scores and/or customized tests to measure chemistry/math skills. Hovey and Krohn were leaders in the investigations and developed the Toledo Chemistry Placement Examination (TCPE). Several universities require TCPE raw scores above 35-50 for entrance into college chemistry. The De- partment of Chemistry at Western Kentucky University (WKU) determined the correlation coefficients between success in college chemistry (grade of A, B, C) and the TCPE total raw scores (0.55). The next-best correlation (0.49) was with the TCPE, part I (math section). The cor- relation of ACT composite score and high school grade- point average with the TCPE scores was extremely poor (0.093-0.296). Average TCPE raw scores (part I, 13.5/20; part II, 11.8/20; part III, 9.4/20; total, 34.7/60); common TCPE questions missed were discussed. GEOGRAPHY Federal land policy and the Wise Use movement. MARY M. SNOW, Department of Geography and Geol- ogy, Western Kentucky University, Bowling Green, KY 42101. Policy makers in the U.S. increasingly are pressured by 54 Transactions of the Kentucky Academy of Science 57(1) the Wise Use movement (WUM) to open all public lands to unconstrained mining, off-road vehicle use, logging, and grazing. While posing as a coalition of grassroots or- ganizations working on behalf of labor and private prop- erty owners, WUM is funded by some of the nation’s larg- est corporations. This paper examines the background of WUM; WUMSs leaders, their perspectives and goals; and major funders of WUM. It analyzes legislation influenced by WUM. Maximizing rural tourism: Horse Cave, Kentucky. RICHARD K. SNOW, Department of Geography and Geology, Western Kentucky University, Bowling Green, KY 42101. The town of Horse Cave, Kentucky, long has been as- sociated with tourism due to the numerous show caves in the area as well as its proximity to Mammoth Cave Na- tional Park. However, the pollution of Hidden River Cave on the town’s main street devastated the tourism industry in 1944. A new sewage treatment plant has helped Horse Cave reclaim some of its former glory. Yet a survey of area businesses reveals that most merchants feel that city of- ficials are not doing enough to attract tourists to Horse Cave. Among recommendations for Horse Cave to maxi- mize its tourism potential are attracting a major motel and restaurant, converting historic homes into bed-and-break- fasts, creating a downtown handicraft market, and estab- lishing a bus service from neighboring Cave City. GEOLOGY Glacial deposition and erosion of the Mile 605 area, Ohio River, Kentucky/Indiana. GRAHAM HUNT, De- partment of Geography and Geosciences, University of Louisville, Louisville, KY 40292. A chronostratigraphic analysis of mainly unconsolidated sediments, soils, and processes of glacial deposition and erosion may be used to reconstruct environmental history of Quaternary age rocks overlying Paleozoic age bedrock in the study area. The exact extent of the Illinoian and Kansan glaciations is unknown in the Louisville area; how- ever, the Kansan drift underlies the Illinoian drift within a few miles of each other. These data suggest that the two ice sheets had about the same southerly extent of advance. Numerous glacial erratics of possible lateral and/or ter- minal moraines are found near the banks of the Ohio Riv- er in northern Kentucky. Sandstone and conglomerate er- ratics are found near grooved, striated, polished, and ex- foliated bedrock of mainly Devonian age carbonates. If reassembled, the erratics would be of several cubic me- ters; the total original weight may be up to 10 tons. Recent drilling in this area indicates a small, U-shaped, and bed- rock-walled valley at depth of Silurian age rocks. The Pleistocene continental deposits at this locality may be di- vided into mappable sequences of lacustrine, outwash, and eolian units overlying the uranium-bearing New Albany Shale. Geologic mapping in population areas is important to evaluate urban mineral deposits of water supply rocks and to help identify and remedy geologic hazards (radon- 222). Radon-222 is commonly released from glacial de- posits. MATHEMATICS A quasi-classical polynomial family. JAMES B. BARKS- DALE JR., Department of Mathematics, Western Ken- tucky University, Bowling Green, KY 42101. This investigation explored some of the properties of a family, H = {h,}_,, of polynomial functions (indexed by degree) created by integrating the Legendre polynomials, {P,}*-o. Specifically, the elements of H are defined by n [1] h(x) =— Cn-1 Jee) Chi, =I where c,_, denotes the leading coefficient of P,_,. In view of Item [1], it may not be surprising that members of H inherit some of the properties and recursive relations en- joyed by the Legendre polynomials. In addition to these “expected” properties, the elements of H possess other curious and interesting attributes, which include the fol- lowing: (a) H is the unique family of polynomials such that each h, is monic, has (x? — 1) as a factor, and has only real, distinct zeros in [—1, 1] coinciding with the inflection point abscissas; and (b) H is a quasi-orthogonal family of vectors in the sense that |n — m| € {0, 2} implies 0 = (h,,, h,,) = JL, h,(t)h,,(t) dt. PVM and parallel computation. KEITH ROE, Depart- ment of Mathematics, Morehead State University, More- head, KY 40351. PVM (Parallel Virtual Machine) is a message-passing computer language. In this project, PVM was used to par- allelize a program, solving a problem of particle transport theory in two dimensions by the invariant imbedding method. This is a computationally intensive large-scale problem, which implies it may be a problem best solved by parallel methods. This program was run on a cluster of HP 9000 workstations and also on a Convex Exemplar machine. The matrix-multiply routine was found to be us- ing 90% of CPU time. This, therefore, was the module that was parallelized. The method for parallelizing the ma- trix multiply was to set up child processes on the other nodes of the cluster and divide the dot product compu- tations among them. Next the program was timed to de- termine if this was an efficient method of solving this problem. Although the parallel version was consistently slower than the serial version, due to message passing overhead, the parallel version consistently grew more ef- ficient as the size of the problem grew. The results of this project demonstrated a basic trend that indicates a parallel version may be efficient for sufficiently large problems. Some concerns in axiomatic set theory. ANDREW MARTIN, Department of Mathematical Sciences, More- head State University, Morehead, KY 40351. The publication of Paul Cohen’s A Minimal Model for Set Theory (1963) and The Independence of the Contin- uum Hypothesis (1964) demonstrated two cases of set the- Abstracts, 1995 Annual Meeting 55 oretic statements (the axiom of choice and the generalized continuum hypothesis) that he showed could not be proved or disproved from the Zermelo-Fraenkel axioms. His method of forcing has since been elegantly refined and developed to prove the undecidability of many other set theoretic statements. It was the view of Goedel that there are other basic principles yet undiscovered that will enable basic set theoretic to be decided. This expository paper reviewed some candidates, in particular Martin’s ax- iom, proper forcing axiom, diamond, projective determi- nacy, and some large cardinal axioms. When is a partially ordered set well ordered? AN- DREW MARTIN,* Department of Mathematical Sci- ences, Morehead State University, Morehead, KY 40351; ALEXANDER ABIAN, Department of Mathematics, lowa State University, Ames, IA 50011. Let S be a subset of a partially ordered set P. We call an element i of P an immediate successor of S if and only if i is a strict upper bound of S and no element of P less than i is a strict upper bound for S. A subset PI of P is called pseudo-inductive provided that for every subset A of PI, if A has immediate successors in P, then at least one of these immediate successors must be an element of PI. We have proved that well-ordered sets can be char- acterized in terms of their pseudo-inductive subsets, in the following sense. THEOREM: A partially ordered set is well-ordered if and only if it has no pseudo-inductive proper subset. MOLECULAR & CELL BIOLOGY Analysis of alpha-fetoprotein enhancer III activity in transgenic mice. JERHONDA M. BRYANT and BRETT T. SPEAR, Department of Microbiology and Immunology, University of Kentucky College of Medicine, Lexington, KY 40536. Alpha-fetoprotein (AFP), a 70,000 dalton glycoprotein, is the major serum protein in the developing mammalian fetus. AFP functions primarily as a transport protein; it may also protect the developing fetus from the maternal immune system. AFP is synthesized at high levels in the liver and the yolk sac and at low levels in the gut. AFP transcription decreases ca. 10,000 fold after birth; this re- pression is reversible as the AFP gene can be reactivated during liver regeneration and in certain cancers. Our lab studies aspects of AFP gene regulation. Three distinct en- hancers, EI, EIT, and EIII, are located upstream from the AFP gene. Previous studies in our lab have identified binding sites for HNF-3 and C/EBP within Enhancer III. HNF-3 and C/EBP sites are liver-specific transcription factors. To further investigate the role of the HNF-3 and C/EBP binding sites, we have altered these sites by PCR- mediated site-directed mutagenesis. Four constructs were produced, the wild type ETI, EIII lacking the HNF-3 site, EII lacking the C/EBP site, and EIII lacking both sites. These four fragments are currently being ligated to a promoter fused to the lacZ reporter gene. The con- structs will be analyzed in tissue culture cells and trans- genic mice. The effects of these mutations on EIII activity will be determined by measuring lacZ activity. We predict that each single mutation will reduce EI activity and that the double mutant will completely eliminate EIII activity. Antibody response and histopathology of Trypanosoma cruzi infection in mice held at elevated temperature. AH- MED A. ARIF and CHERYL D. DAVIS, Department of Biology, Western Kentucky University, Bowling Green, KY 42101. Highly susceptible C3H mice survive an otherwise le- thal infection with Trypanosoma cruzi when held at an elevated temperature of 36°C. The body temperature of the mice increases 3° to 4°C and the mice experience de- creased parasitemia and enhanced parasite-specific and nonspecific immune responses. The present study was de- signed to determine whether heat shock proteins (hsps) of the parasite might be playing a role in this phenome- non. Antibody responses to parasite hsps were analyzed using metabolic labelling with *S methionine/cysteine fol- lowed by SDS PAGE and autoradiography. The results showed that by day 15, serum from RT-infected mice rec- ognized proteins of approximately 93 kDa, 83 kDa, and 66 kDa, while in mice held at 36°C the major proteins recognized were 93 kDa and 83 kDa. By day 25, a high MW band of ca. 183 kDa was recognized by serum from mice held at RT and 36°C. A stronger response to the 93 kDa, 83 kDa, and 66 kDa proteins was seen in RT-in- fected mice as compared to 36°C infected mice. By day 35, another high MW protein of approximately 192 kDa was recognized by sera from both RT and 36°C infected mice. Strong reactivity with the 183 kDa, 93 kDa, 83 kDa, and 66 kDa proteins was seen in both groups of mice on day 35. By day 45 of infection, serum from 36°C infected mice recognized proteins of ca. 183 kDa, 93 kDa, 83 kDa, and 66 kDa more intensely than in the RT-infected mice group. In addition, a low MW protein of approximately 27 kDa was recognized only by serum from 36°C infected mice. Comparative analysis of plant mitochondrial small sub- unit ribosomal RNA. CANDACE L. GLENDENING,* CRAIG A. TUERK, and GERALD DEMOSS, Depart- ment of Biological and Environmental Sciences, More- head State University, Morehead, KY 40351. The small subunit of the ribosomal RNA has long been used as a tool in determining phylogenetic relationships. This project explored a region of the mitochondrial small subunit ribosomal RNA (MSrRNA) found only in the plant mitochondrial genome referred to as Variable Re- gion 7 (V7). The V7 region of cauliflower and radish, members of the Brassicaceae, was amplified with the poly- merase chain reaction, cloned into the expression vector PBSSK+, and sequenced. There was only a 0.3% se- quences difference between cauliflower and radish V7 re- gion, thus suggesting that there will not be sequence vari- ation in this region within a plant species. Cauliflower and radish V7 sequence was compared with the six other se- 56 Transactions of the Kentucky Academy of Science 57(1) quences of plant MSrRNA V7 region available: wheat, corn, oats, evening-primrose, soybean, and lupine. Based on percent difference between the V7 region sequences, a phylogenetic tree was constructed that supports place- ment of these species in Cronquist’s morphologically based phylogenetic tree of flowering plants. Determination of behavior of antigen-specific T-cells following immunization to an immune privileged site. DA- VID PEYTON,* RITA EGAN, and JEROLD WOOD- WARD, Department of Microbiology and Immunology, University of Kentucky, Lexington, KY 40502. The immune system cannot react the same way in all situations. Sites such as the eye and central nervous sys- tem would be harmed or destroyed if they received the same immune response to antigen that most other sites in the body receive. These special sites, known as immune privileged sites, can be studied using adoptively trans- ferred mice and fluorescent antibody stained T-cells. Adoptively transferred mice are created by intravenous injection of a small population of ovalbumin-specific T- cells from a transgenic mouse into a non-transgenic mouse. The adoptively transferred mice receive an oval- bumin injection into the anterior chamber of the eye. To monitor the activity of the T-cells, the lymph nodes and spleen from the injected mice are removed and the T- cells from these organs are isolated. Next, the T-cells are stained with two separate fluorescent antibodies: one spe- cific for the ovalbumin-specific T-cells and one that binds to all CD4+ T-cells. Using a flow cytometer to measure the amount of staining, the T-cells specific for ovalbumin can be distinguished, quantified, and compared to non- ovalbumin-specific T-cells. Data from this study indicated that there was a local draining lymph node response in the cervical and submandibular nodes following antigen administration in the eye. These data also suggest that there, is not a response in the venous circulation, which would have been indicated by a population of ovalbumin- specific T-cells in the spleen. Determinations of residues involved in interchain for- mation and stability of hamster aspartate transcarbamy- lase. PANNA PATEL* and JEFFREY N. DAVIDSON, Department of Immunology and Microbiology, University of Kentucky, Lexington, KY 40502. Aspartate transcarbamylase (ATCase) is an independent monofunctional enzyme in Escherichia coli. In hamster, ATCase is one of the three domains of the multifunctional CAD protein. CAD, a multimer of identical subunits, is named after its three enzymes: carbamyl-phosphate syn- thetase, ATCase, and dihydroorotase. Together these three enzymes catalyze the first three steps of de novo pyrimidine biosynthesis in which ATCase specifically cat- alyzes the second step. In E. coli the enzyme is a trimer of catalytic chains; adjacent catalytic subunits form shared active sites. The objective here was to identify residues of the hamster ATCase not directly involved in the shared active site but involved in stabilizing the interactions be- tween CAD monomers. To achieve this aim, an expression plasmid encoding the ATCase domain of hamster CAD was expressed in E. coli, and five changes were introduced by site-directed mutagenesis: Phe286 — Alanine, Phe286 — Tyrosine, Arg287 — Glutamine, Gln288 — Asparagine, and Gln288 — Glutamic acid. Reduced enzymatic activ- ities were observed. Heat treatment of bacterial extracts at 50°C for 5 minutes showed the most drastic effects on enzymatic activities of Phe286 — Ala and Phe286 —> Tyr. For these two mutants, the hamster CAD cDNA was cloned into the pMALc2 expression vector, a special vec- tor that permits one step purification of the recombinant protein. After purifying pMALc2-Phe286 — Ala protein, its multimeric structure was determined by native gel electrophoresis. Because the protein migrated to a smaller pore size than the original wild-type ATCase, this indicat- ed that the protein has an altered multimeric structure, suggesting that Phe286 plays a role in CAD structure and that substitution at this point can disrupt the multimer. Dietary activation of the hypothalamic-pituitary-adrenal axis through the histaminergic system. AMY TIU,* HOL- LY BUNDRANT, and L. PRESTON MERCER, Depart- ment of Nutrition and Food Science, University of Ken- tucky, Lexington, KY 40506. During protein deficiency, increased histaminergic ac- tivity increases adrenocorticotropic hormone (ACTH) and corticotropin releasing factor (CRF), which may lead to increased levels of corticosterone in rats and cortisol in humans. Hypercortisolemia is a condition found in cases of anorexia nervosa (AN). Corticosterone levels were ex- amined in 19 male and 19 female Sprague-Dawley rats fed diets containing either 25% casein or 1% casein. One group of seven male rats and six female rats was fed a 1% casein diet and received a subcutaneous injection of dox- epin, a central histamine receptor antagonist (10 mg/kg; DOX). Plasma from the rats were then analyzed by a cor- ticosterone solid phase !*°I radioimmunoassay. The mean levels of corticosterone in each treatment group (25% ca- sein, 1% casein, and 1% casein + DOX) of males were 1618.54 nmol/liter, 1236.45 nmol/liter, and 1115.22 nmol/ liter, respectively. The mean levels of corticosterone in each treatment group (25% casein, 1% casein, and 1% casein + DOX) of females were 1969.88 nmol/liter, 1491.53 nmol/liter, and 1512.89 nmol/liter, respectively. All groups were significantly different from each other (P = 0.05). Rats injected with doxepin showed improved food intake, weight gain, and efficiency. Even though no statistical differences were found in comparison of corti- costerone levels between gender and between treatment groups within gender, the results suggest that corticoste- rone levels may be influenced by protein deficiency and the histaminergic system. Effects of lead acetate on host susceptibility to Trypanosoma cruzi. TABITHA ELLIS and CHERYL DA- VIS, Department of Biology, Western Kentucky Univer- sity, Bowling Green, KY 42101. Abstracts, 1995 Annual Meeting Di Lead has been found to be an immunotoxicant that sup- presses the resistance of a host organism to infection. C57BI/6] mice, which are not highly susceptible to the Brazil strain of Trypanosoma cruzi, were chosen to deter- mine if and how lead would affect their immune response upon infection. The mice were acclimated for 2 weeks on deionized water and rodent chow, prior to lead treatment. They were divided into four groups and were given oral dosages of lead acetate ranging from 0 to 1,000 p.p.m. in their drinking water for 3 weeks. On day 35 of the exper- iment, the mice were injected with 10‘ blood form try- pomastigotes of Trypanosoma cruzi; the lead treatment continued. Beginning at 14 days post infection, parasitem- ias were conducted two times weekly. The mean peak par- asitemias for each group were as follows: 0 p.p.m.—6.49 x 10°, 10 p.p.m.—9.61 X 10°, 100 p.p.m—1.31 X 107, and 1000 p.p.m.—3.62 x 10°. The concentrations of lead in blood in the first replicate of the experiment were: 0 p-p.m.—).34 g/liter, 10 p.p.m—19.72 ypg/liter, 100 p-p.m.—16.36 g/liter, and 1000 p.p.m.—204.41 g/liter. Although no increased mortality was observed in lead- treated mice, the results of this study suggest that lead does enhance the susceptibility of infected mice, resulting in higher parasitemias. Effects of viral infection on insect immune cell func- tion. AARON CAMERON#* and BRUCE A. WEBB, De- partment of Entomology, University of Kentucky, Lexing- ton, KY 40502. The parasitoid wasp Campoletis sonorensis injects cer- tain immunosuppressive factors when it preys upon its lepidopteran host. One of these factors is the polydnavi- rus, the only known DNA virus with a segmented genome. The polydnavirus has proven to have adverse effects upon the host; in particular, suppression of the cellular immune response. A procedure for collecting insect hemolymph and purifying hemocyte cell populations has been opti- mized for use in determination of viral gene expression in parasitized insect RNA. The viral gene of interest has been partially sequenced, creating a sequence-specific tag, allowing for its identification and characterization into the rep-gene family. A Northern has been performed on RNA extracted from parasitized insects to examine phases of viral gene expression post-injection. Research of this na- ture is pertinent to both agriculture and human immu- nology. Electrophoretic analysis of carotid plaque lipoproteins. AMANDA R. TATRO* and ROBERT A. LODDER, Col- lege of Pharmacy, University of Kentucky Medical Center, Lexington, KY 40536. Determining the number of proteins present in an ath- eroma cell and the roles played by these proteins will be essential in the future to diagnosis and treatment of ath- erosclerosis. In vitro plaque electrophoresis and in vivo near-IR spectrometry are used to provide qualitative and quantitative information on the proteins involved in ca- rotid atherosclerotic lesions. The information garnered from electrophoresis is used to test the hypothesis that particular proteins in combination show significant cor- relation to patient medical history and lesion pathology. Bruit, from medical history, and fibrous cap, from pa- thology reports, are shown to involve similar concentra- tions of the same proteins. Near-IR spectra of these pro- teins indicate that all but one of the protein factors char- acteristically found are probable lipoproteins. In addition, the concentration of the 18 kDa protein is related to lesion size, which in turn has been related to severity of disease. Duplex ultrasound measurements are used to determine lesion size and are compared to protein concentrations determined by electrophoresis to demonstrate a relation- ship between the concentration of the 18 kDa protein and the size of the lesion. Expression of human NAD*‘-dependent 15-hydroxy- prostaglandin dehydrogenase in yeast. GARY HALL,* MARK ENSOR, and HSIN-HSIUNG TAI, Department of Medicinal Chemistry and Pharmaceutics, College of Pharmacy, University of Kentucky, Lexington, KY 40536. Prostaglandins are fatty-acid-derived compounds exhib- iting a broad spectrum of biological activities. Once pro- duced, they are rapidly metabolized and inactivated. The first step of their metabolism is catalyzed by NAD*-de- pendent = 15-hydroxyprostaglandin — dehydrogenase (PGDH). For this reason PGDH is considered to be the key enzyme controlling inactivation of the prostaglan- dins. However, the nature of the interaction of the pros- taglandins with the active site of the enzyme is not known. A future goal is to determine the three-dimen- sional structure of PGDH by x-ray crystallograpy. De- termining the structure will aid in determining the cat- alytic mechanism of the enzyme as well as how phar- macological agents interact to exert their effects. The levels of PGDH expression in recombinant mammalian and bacterial systems make it difficult to produce and purify PGDH in sufficient amounts for x-ray crystallo- graphic studies. Recently, a yeast expression system (Pi- chia) was developed that should allow for production of recombinant proteins in much higher amounts. The cDNA for PGDH was inserted into the yeast expression plasmid (pHIL-D2), which was then used to transform the yeast Pichia pastoris. A recombination event oc- curred within the yeast between the yeast alcohol oxi- dase (AOX1) gene sequences contained in the plasmid and the AOX1 gene in the yeast chromosome. This re- combination results in replacement of the yeast chro- mosomal AOXI gene with the PGDH cDNA. The PGDH cDNA is then under the control of the AOX1 promoter. This promoter is regulated by the presence of methanol. When methanol is present, very high levels of expression from the AOXI promoter occur. Maximum expression of PGDH was obtained after 2 days of in- duction with methanol. Levels of expression were 25 times higher than that obtained in Escherichia coli. Ex- pression of PGDH using this yeast system will greatly 58 Transactions of the Kentucky Academy of Science 57(1) facilitate the purification of the enzyme for x-ray crys- tallography. Identification of cytochrome P450 monoxygenase en- zyme genes in corn. JOHN DAVID GREULICH*, MI- CHAEL BARRETT, NICHOLAS POLGE, and JOHN RAWLS, Department of Agronomy, University of Ken- tucky, Lexington, KY 40546. Most organisms contain cytochrome P450 monoxygen- ase enzymes (P450), which are involved in various biolog- ical reactions such as the production of secondary metab- olites and the detoxification of environmental pollutants and xenobiotics. Presently, two inbred lines of corn have been found to express and not express P450 genes, which are responsible for the detoxification of particular pesti- cides. Further, these genes are found to be inducible and non-inducible with treatment by certain chemicals. Poly- merase chain reaction (PCR) has been used previously to isolate partial sequences of corn P450 genes. The con- struction of a cDNA library is necessary to obtain full- length sequences. A cDNA library was produced from one inbred line using a library construction kit to make cDNA from isolated mRNA. This library and another produced commercially will be screened by P450 probes centered around a conserved heme binding region characteristic of P450s. Clones showing homology to probes will be se- quenced in an attempt to obtain full-length sequences of P450 genes of interest. Comparison between the two lines will give insight into the manner in which the p450 genes are regulated. Further study into the regulation of p450 genes would require construction of a genomic DNA li- brary. Additionally, the inducible line could be studied in greater detail, as it shows multiple substrate activity. This could be due either to a single enzyme acting upon a variety of substrates or to many enzymes with genes close- ly associated on the same chromosome. These would not show segregation with classical genetic analysis. Interactions of the tobacco vein mottling virus’s mutant cylindrical inclusion and nuclear inclusion B_ protein. BETH SMITH* and ARTHUR HUNT, Department of Agronomy, University of Kentucky, Lexington, KY 40508. The tobacco vein mottling virus (TVMV) is a single- stranded RNA plant potyvirus containing six classified coding regions: the Cylindrical Inclusin gene, the Nuclear Inclusion A gene, the Nuclear Inclusin B gene, the Helper Component gene, the Coat Protein gene, and the P1 gene. The coding regions of interest, the CI-and NIb genes, both have putative roles in viral replication as a helicase and polymerase, respectively. Previous research identified moderate protein-protein interactions between the CI and NIb proteins. To determine if the active sites of these proteins were responsible for the identified interactions, CI and Nib fusin proteins were designed to contain mu- tations within regions of these proteins’ putative active sites. The NIb gene expresses a 58 kDa protein that has been implicated in viral replication as an RNA-dependent RNA polymerase. The most conserved of the potyviral proteins, the NIb protein’s putative active site is a GDD domain. The CI gene codes for a protein 70 kDa known as the cylindrical inclusion protein. Cylindrical inclusion proteins aggregate within the cytoplasm of plant cells in- fected by the TVMV virus to form pinwheel-like shaped structures. The CI protein has been included in a pro- posed superfamily of proteins that have putative ATP-de- pendent helicase activity. There are six regions within the CI domain that contain significant sequence homology with the other proteins included in the proposed helicase superfamily. A CI mutant containing a point mutation in region VI was used to determine if one of the putative active sites for helicase activity were involved in the pro- tein-protein interactions identified, while a NIb clone de- signed with a point mutation in the GDD domain was provided by Dr. Tom Pirione’s laboratory. The data sug- gest that the mutations in the putative active sites of the CI and Nib proteins insignificantly affected the identified interaction. Lipoprotein determination in single cells by near-infra- red spectromicrography. JENNIFER L. MOSES* and ROBERT A. LODDER, College of Pharmacy, University of Kentucky Medical Center, Lexington, KY 40536. A near-infrared indium-antiminide (InSb) focal plane array camera is being used to collect images of carotid plaques during carotid endarterectomies. The excised plaques are later analyzed for lipoprotein content by ul- tracentrifugation and gel electrophoresis. The results from the electrophoresis indicate a correlation between the size of atherosclerotic plaque and the concentration of a 93 kDa protein in the plaque, and plaque size is correlated to stroke. Further study of the 93 kDa protein may lead to a drug to block its receptor in plaque cells. This report describes tests of two hypotheses: (1) that a near-infrared platinum-silicide (PtSi) charge-coupled device camera provides better in vivo spatial resolution and more func- tional pixels on a plaque image than the InSb focal plane array camera, with sufficient signal-to-noise ratio for mac- roscopic lipoprotein determination in individual plaque cells, and (2) that the oxLDL 93 kDa protein is trans- ported into plaque cells from serum by infiltrating mac- rophages. The InSb camera has more dead pixels and a lower spatial resolution than the PtSi camera. The stan- dard error of estimate and standard error of prediction for analytes using the InSb camera and PtSi camera were compared in the laboratory using prepared samples to cre- ate a calibration curve. The increased spatial resolution of the PtSi camera makes it easier to select spectra from microscopic sections made from the excised plaque. The spectra collected by attaching the PtSi camera to a micro- scope are also compared to a visual picture of a stained slide from the same plaque for reference. Nicotine metabolite contribution to the central nervous system effect of nicotine. SUSAN MOORE,* WENDY SHAW, SUSAN BUXTON, and LINDA DWOSKIN, De- Abstracts, 1995 Annual Meeting 59 partment of Pharmacology and Experimental Therapeu- tics, University of Kentucky, Lexington, KY 40536. Dopamine is believed to play a major role in the rein- forcing properties of drugs of abuse, including nicotine. The purpose of this research was to determine if the CNS effects of nicotine exposure are due in part to the nicotine metabolite-induced activation of the dopaminergic recep- tors. The nicotine metabolites—nornicotine, cotinine, and norcotinine—have been shown to be in the CNS following nicotine exposure. These nicotine metabolites were inves- tigated to determine their ability to evoke [‘H|dopamine release from rat striatal slices and to inhibit [*H ]dopamine uptake in rat striatal synaptosomes. [*H|dopamine release evoked by nornicotine was found to be equivalent to re- lease evoked by nicotine, whereas cotinine was less effec- tive, and norcotinine was ineffective. Further, the evoked release was not the result of inhibition of dopamine up- take. Thus, these nicotine metabolites activate dopami- nergic receptors indirectly, and therefore may contribute to the abuse liability of tobacco smoking. Role of EGR-1 in prostate cancer cell programmed cell death. REZA F. SAIDI* and VIVEK M. RANGNEKAR, Department of Surgery, University of Kentucky, Lexing- ton, KY 40536. Androgens stimulate both normal and malignant growth in prostate. Androgen-ablation is a standard therapy for metastatic prostate cancer. However, this therapy is rarely curative because prostate cancer is heterogeneous in cell content, including both androgen-dependent and andro- gen-independent cells. Our studies indicate that phorbol 12-myristate 13-acetate (PMA) causes death in both an- drogen-dependent (LNCaP) and androgen-independent (PC-3) cells. PMA-inducible death of LNCaP and PC-3 cells exhibited molecular features indicative of pro- grammed cell death (apoptosis) particularly oligonucleo- some-length DNA fragmentation. Our data showed that this process was linked to an increase in expression of EGR-1, a zinc finger transcription factor. Functional stud- ies using dominant-negative mutant of EGR-1 and anti- sense oligomers indicate that EGR-1 plays an important role in prostate cancer cell apoptosis. Role of NAD as a P53 modulator and chemopreventive agent. MANDALA V. WILSON*, ARNOLD C. HUANG, and ELAINE L. JACOBSON, Department of Clinical Sciences, University of Kentucky, Lexington, KY 40506. The primary metabolite of the vitamin niacin is NAD. Nicotinamide, nicotinic acid, and tryptophan are reported precursors of NAD; however until now studies have not been conducted to determine whether they also serve as precursors in breast epithelial cells, and the quantitative requirements of precursor to metabolite are unknown. Recently a new function of NAD was shown to be as a substrate in synthesis of ADP-ribose polymers when can- cer-causing chemicals induce DNA strand breaks. These polymers function in recovery of DNA damage. About 15% of the western world is predicted to be niacin defi- cient. HME cells are principle precursor cells for breast cancer and are at particular risk for niacin deficiency due to periodic cycles of cell division. DNA damage is an im- portant step in the process of developing cancer cells. Cel- lular responses to DNA damage such as DNA repair limit development of cancer cells. We have developed a method for extracting and measuring both oxidized and reduced NAD and NADP from any tissue in a single step; this allowed us to analyze cells for niacin status much faster than by previous methods. We have also been able to reg- ulate NAD content in HME cells by growing them for several population doublings in nicotinamide free medi- um. Cellular NAD modulates the levels of p53, a tumor suppressor gene product. pos assists in responses to DNA damage by stopping cell cycle progression in order for repair to take place. Below-normal p53 response is asso- ciated with an increased frequency of tumors in animals and humans; mutations in p53 are the most widely rec- ognized mutation found in tumors. We have shown that the p53 levels in cells with depleted nicotinamide levels are lower than in cells with normal nicotinamide levels. Since p53 levels are known to be elevated following DNA strand breaks, the effect of niacin modulation are now being studied using oxidative stress as a DNA damaging agent. These final studies should establish the quantitative requirements for niacin and NAD in the p23 response to DNA damage. Specific role of 92-kDa Type IV collagenase in meta- static tumor cells) DAVID DEREMER,* STEPHEN ZIMMER, and CONNIE ZIMMER, Department of Im- munology and Microbiology, University of Kentucky, Lex- ington, KY 40502. Studies have shown that metalloproteinases, specifically 92-kDa type IV collagenase, may mediate metastasis. Ac- tivity of 92-kDa type IV collagenase was evaluated in two distinct cell types: a non-metastatic C1300 neuroblastoma and a non-metastatic HKBI fibroblast. The C1300 and HKBI1 cell lines were evaluated by zymogram and western blot analyses for the presence of 92 kDa type IV colla- genase. Those cells expressing high levels of 92 kDa type IV collagenase were subcutaneously injected in nude mice. In each case, 92 kDa type IV collagenase correlates with metastasis but is not the only molecule down regu- lated. In addition to 92 kDa type IV collagenase, devel- opment of metastatic potential is correlated with sup- pressed CD44V and stomelysin, as well as an increase in nm 23-H1 (a putative metastasis-suppressor gene). To date it is not known whether the 92 kDa type IV colla- genase, by itself, is sufficient to establish a malignant phe- notype. Targeting the poly(ADP-ribose) glycohydrolase gene in cancer chemotherapy. CHRISTOPHER D. WATT* and ELAINE L. JACOBSON, Department of Clinical Sci- ences, University of Kentucky, Lexington, KY 40506. One of the most rapid responses to DNA strand breaks is the formation of complex ADP-ribose polymers from 60 Transactions of the Kentucky Academy of Science 57(1) the substrate nicotinamide adenine dinucleotide (NAD). The modification of chromatin protein by these polymers is known to be a critical step in cellular recovery. Inhibi- tion of this metabolism enhances the cytotoxicity of agents that generate DNA strand breaks. Our target for inhibi- tion of this metabolism enhances the cytotoxicity of agents that generate DNA strand breaks. Our target for inhibi- tion, in this multienzymatic process, in poly(ADP-ribose) glycohydrolase (PARG), which catalyses degradation of ADP-ribose polymers following DNA repair. Prior re- search has shown adenosine diphosphate (hydroxymethyl) pyrrolidinediol (ADP-HPD) to be a potent and selective inhibitor of PARG activity in vitro. We investigated the general cytotoxicity of the drug as well as its ability to amplify effects of N-methyl-N’-nitro-N-nitroguanidine (MNNG), and oxidative DNA damaging agent, in cultured animal cells. Based on colony forming and MTS cell pro- liferation assays, ADP-HPD is slightly toxic to undamaged cells. The cytotoxicity of MNNG is, however, amplified up to 10-fold when MNNG is applied in the presence of ADP-HPD. To verify the mechanism of this biological re- sponse, ADP-ribose polymer levels must be measured and compared in ADP-HPD treated and untreated cells fol- lowing DNA damage. Initial experiments to quantify ra- diolabeled polymers in damaged cells showed that ADP- HPD decreased the amount of polymer formed. The ef- fect of antisense mRNA targeting the PARG gene will provide confirmation of this observation. Our preliminary data suggest PARG is indeed a likely target for the mod- ification of responses to chemotherapy. T-cell responses of the channel catfish, Ictalurus punc- tatus, to outer membrane proteins of Edwardsiella ictal- uri. BARRY R. HAMILTON* and GEOFFREY W. GEARNER, Department of Biological and Environmen- tal Sciences, Morehead State University, Morehead, KY 40351, Although the immune system of the channel catfish has been studied for many years now, the role of the channel catfish T-lymphocyte is still poorly understood. There are several efforts underway to develop a vaccine against Ed- wardsiella ictaluri, the causative agent of enteric septice- mia of catfish. Current vaccines can induce a specific an- tibody response that confers some protection but is not 100% effective. An understanding of the cellular, as well as the humoral, immune response is needed to develop an effective vaccine. In this project, channel catfish T- lymphocyte mitogenic responses to outer membrane pro- teins of E. ictaluri was examined. Ten micrograms of pro- tein was sufficient to induce a significant proliferative re- sponse. There was a significant proliferative response to E. ictaluri outer membrane proteins that occurred in the range of 55 to 66 hours in pre-exposed channel catfish and in the range of 66 to 78 hours in control catfish. Tissue-specific regulation of renin angiotensin compo- nents by cold exposure. JERRI R. DYER,* ANN TRAN, and LISA A. CASSIS, Division of Pharmacology and Ex- perimental Therapeutics, College of Pharmacy, University of Kentucky, Lexington, KY 40536. Angiotensin II (AI) has been found to be a major pep- tide factor regulating blood pressure. Recent studies have shown that AII may be formed independently of the blood stream in a variety of tissues. One location for this for- mation is rat interscapular brown adipose tissue (ISBAT). ISBAT, a highly metabolic tissue, is regulated primarily by the sympathetic nervous system. Thus, an increase in IS- BAT metabolism induced by cold-exposure is hypothe- sized to result in increased production of AIT. This study will determine if increased metabolism regulates AIT pro- duction in ISBAT. Molecular characterization of the syn- thetic components of the RAS will be determined in IS- BAT following different periods of cold-exposure. An in- crease in mRNA expression for angiotensinogen and an- giotensin converting enzyme in ISBAT and not in liver or kidney will provide evidence suggesting tissue-specific regulation of AIT synthesis. Use of a genetic technique to identify yeast components of the splicing apparatus. GUY EAKIN* and BRIAN RY- MOND, T. H. Morgan School of Biological Sciences, Uni- versity of Kentucky, Lexington, KY 40502. The primary RNA transcripts (pre-mRNA) of most eu- karyotic genes contain non-coding sequences (introns) within the protein coding segments. Removal of these in- trons is, in most instances, imperative for the RNA to direct protein synthesis. The splicing of pre-mRNA to yield mature mRNA is a multi-step process involving sev- eral intermediates. As DNA is transcribed within the nu- cleus, the introns are excised from the pre-mRNA tran- script by the spliceosome, a complex enzyme consisting of 50-100 polypeptides and 5 small nuclear RNAs (snRNAs). Commitment of the pre-mRNA to the splicing pathway is dependent on stable association of the pre-mRNA mole- cule with the U1 small nuclear ribonucloprotein particle (consisting of the Ul snRNA and associated polypetides). In addition, our lab has previously shown that, in Saccha- romyces cerevisiae, the PRP39 gene product (PRP39p) is essential for stable association of the Ul snRNP with the pre-mRNA transcript. The focus of this project was to identify other gene products that genetically interact with the PRP39p by use of a genetic technique, high copy number médiated suppression. A library of self-replicating plasmids containing random segments of the yeast ge- nome was used to transform a temperature sensitive (ts) strain of yeast, ts307. Of ca. 30,000 transformants 20-40 colonies were obtained wherein the mutant ts prp39 con- ditional lethal defect was relieved. Preliminary character- ization of 13 plasmids suggest that, in addition to isolation of wild-type PRP39, other genes that interact with PRP39p were represented. On-going studies will prove the possible role of novel genes in the pre-mRNA splicing process. PHYSICS Asteroid orbital determinations based on CCD obser- vations. RICHARD HACKNEY,* KAREN HACKNEY, Abstracts, 1995 Annual Meeting 61 ROGER SCOTT, and JIM PARVIN, Western Kentucky University Astrophysical Observatory, Department of Physics and Astronomy, Western Kentucky University, Bowling Green, KY 42101. Students in observational astronomy courses, or who otherwise have an interest in participating in observational project experiences, can use CCD images of asteroids as a basis for determining the approximate distances of the asteroids from the sun. We describe a student activity us- ing CCD observations of a selected asteroid made ca. | hour apart, along with the necessary derivations to use measurements of the change in position of an asteroid near opposition to evaluate the apparent angular velocity and to deduce the size of the orbit, assuming low orbital eccentricity. Students can make the determinations using either CCD images that they acquire themselves with available equipment, or using images recorded using the 24-inch reflector at the Western Kentucky University As- trophysical Observatory. Heaviside operational exact analytical solutions of the time-dependent Hamilton-Jacobi equation initial-value- problem defined by model real/complex rheonomic Ham- iltonians. VALENTINO A. SIMPAO, 108 Hopkinsville St., Greenville, KY 42345. Exact analytical solutions of the initial-value-problem for the time-dependent Hamilton-Jacobi equation defined by broad classes of model real/complex rheonomic Ham- iltonians, have recently been discovered via a formal Heaviside operational scheme. Heaviside operational exact analytical solutions of the time-dependent Schrodinger equation initial-value-prob- lem defined by model real/complex rheonomic Hamilto- nians. VALENTINO A. SIMPAO, 108 Hopkinsville St., Greenville, KY 42345. Exact analytical solutions of the initial-value-problem for the time-dependent Schrodinger equation defined by broad classes of model real/complex rheonomic Hamilto- nians have recently been discovered via a formal Heavi- side operational scheme. NASA-Kentucky partnership opportunities for space-re- lated science, technology, and economic development. KAREN HACKNEY,* RICHARD HACKNEY, and ROG- ER SCOTT, Kentucky Space Grant Consortium and NASA EPSCoR Program, Department of Physics and Astronomy, Western Kentucky University, Bowling Green, KY 42101. NASA has established partnerships with Kentucky and its universities for the purpose of involving faculty and students in space-related research, as well as technolog- ical and economic development. We describe current opportunities in the Kentucky Space Grant Consortium for undergraduate scholarships and graduate fellowships for students in mentored projects. Funding opportuni- ties for the development of space-related projects and activities to support the teaching of space science and related disciplines are outlined. Progress of existing pro- jects and new opportunities in the Kentucky NASA EPSCoR Program are discussed. Novel analytically soluble model multi-particle interact- ing time-dependent classical dynamical systems. VAL- ENTINO A. SIMPAO, 108 Hopkinsville St., Greenville, KY 42345. Developed herein is a theoretical model of multi-par- ticle interacting time-dependent classical dynamics, which admits analytical solution. The program is based upon my recent studies in analytical solution schemes for the time- dependent Hamilton-Jacobi initial-value-problem, defined by broad classes of model real/complex time-dependent Hamiltonians and initial conditions. The objective is achieved by particularizing the cited solution schemes to generate and solve the proposed model system. Meeting the objective results in enhanced theoretical and compu- tational capabilities (i.e., algorithms) for analysis of actual physical multi-particle interacting time-dependent classi- cal dynamical systems. Novel analytically soluble model multi-particle interact- ing time-dependent quantum dynamical systems. VAL- ENTINO A. SIMPAO, 108 Hopkinsville St., Greenville, KY 42345. Developed herein is a theoretical model of multi-par- ticle interacting time-dependent quantum dynamics, which admits analytical solution. The program is based upon my recent studies in analytical solution schemes for the time-dependent Schrodinger initial-value-problem, defined by broad classes of model real/complex time-de- pendent Hamiltonians and initial conditions. The objec- tive is achieved by particularizing the cited solution schemes to generate and solve the proposed model sys- tem. Meeting the objective results in enhanced theoretical and computational capabilities (i.e., algorithms) for anal- ysis of actual physical multi-particle interacting time-de- pendent quantum dynamical systems. Opportunities for minority students in astrophysics us- ing robotic observing facilities in the Center for Auto- mated Space Science. ROGER SCOTT,* CHARLES McGRUDER, KAREN HACKNEY, and RICHARD HACKNEY, Center for Automated Space Science, De- partment of Physics and Astronomy, Western Kentucky University, Bowling Green, KY 42101; MICHAEL BUS- BY, Center for Automated Space Science, Center of Ex- cellence in Information Systems Engineering and Man- agement, Tennessee State University, Nashville, TN 37209. NASA has awarded funding to support a University Re- search Center titled the Center of Automated Space Sci- ence (CASS). The program is centered at Tennessee State University (TSU) under the direction of Dr. Michael Bus- by, with major components at Western Kentucky Univer- sity (WKU) and at South Carolina State University (SCSU). Objectives of the program include establishment of research programs using remotely operated ground- 62 Transactions of the Kentucky Academy of Science 57(1) based telescopes and NASA missions, and development of procedures for dynamic scheduling and conducting astro- physical observations using automated instrumentation. The multi-faceted program will provide opportunities for involving under-represented minority students in prepar- ing for careers in space-related sciences. Research oppor- tunities at TSU include developing and managing robotic observing facilities and using the instruments to study magnetic effects in cool stars, including solar-type stars with starspots and magnetic activity relevant to the solar cycle and its effects on the earth. At WKU, a related theme is the effect of celestial high-energy sources, in- cluding the sun, x-ray sources, and gamma-ray bursters, on the earth’s ionosphere. At SCSU, the studies extend beyond the solar system to understand the physics of the interstellar medium and nebular concentrations within it. Probing still greater distances, far beyond our galaxy, a group at WKU is studying the behavior of active galactic nuclei—energetic objects such as quasars with rapidly varying energy output. Opportunities for minority stu- dents to participate in the development of automated re- mote observing programs, and in astrophysical research, will be described. Spectral classification of B supergiants in the Small Mag- ellanic Cloud with the International Ultraviolet Explorer. RAYMOND C. McNEIL* and ANTHONY L. BORCH- ERS, Department of Physics and Geology, Northern Ken- tucky University, Highland Heights, KY 41099. The spectral classifications of 79 B stars known or sus- pected to be supergiants and members of the Small Mag- ellanic Cloud (SMC) are being estimated by means of vi- sual examination of their ultraviolet spectra. All program spectra are new or archival low-dispersion spectra ob- tained with the short wavelength prime camera (1200 A < d < 2000 A) of the International Ultraviolet Explorer (LUE). The work is being carried out in collaboration with George Sonneborn and Richard P. Fahey of the Goddard Space Flight Center (GSFC) using the computing facili- ties of the IUE Data Analysis Center. The spectral clas- sifications are based on criteria suggested by literature and/or illustrated by sequences of low-dispersion TUE spectra of galactic standards. Archival high-dispersion IUE spectra of additional galactic standards have also been obtained and resampled to approximate the resolu- tion of the low-dispersion spectra. Spectral features se- lected for classification are limited to photospheric ab- sorption lines within the stated range of wavelengths. Pre- liminary classification of 41 SMC spectra suggests that nearly half the program stars may fall outside the range defined by the low-dispersion sequence of BO to B9 ga- lactic supergiants. One-third of such spectra appear to be hotter than BO; the remaining two-thirds appear to be giants or bright giants. Spectral criteria are therefore be- ing carefully re-examined before final program classifica- tions are determined. The galactic center’s annihilation line (0.511 MeV). TODD GRIFFITH,* JUNMEI PAN,* SHAWN BRAD- LEY,* and ANDREW MARTIN, Department of Mathe- matical Sciences, Morehead State University, Morehead, KY 40351. Balloon-borne instruments in the 1970s first detected the galactic center's strong 0.511 MeV spectral line. As 0.511 MeV is the precise energy equivalent to one elec- tron mass, it was generally accepted that this line was the signature of electron-positron annihilation. However, sub- sequent occasional monitoring has shown considerable variation in the flux of 0.511 MeV photons being received. In fact, in the early 1980s the flux dropped to nearly zero. But in the late 1980s the line “turned on” again and has remained at about the same level until now. It now seems generally accepted that there are two sources for the pos- itrons whose annihilation causes this line. One is a com- pact time-variable source (probably 1E 1740.7-2942); the other is a galactic plane component, probably due to neu- tron stars. PHYSIOLOGY & BIOCHEMISTRY Antioxidant influence of platelet aggregation and oxi- dation of low-density lipoprotein in hypercholesterolemic rats. R. W. THURMAN* and D. J. SAXON, Department of Biological and Environmental Sciences, Morehead State University, Morehead, KY 40351. The effects of dietary vitamin E supplementation, an antioxidant, on platelet aggregation and oxidation of low- density lipoprotein (LDL) were investigated. Male rats re- ceived either a normal diet (N), a N diet plus 5% choles- terol (C), or a C diet plus 300 mg vitamin E/kg diet (CE) for 10 weeks. There were no differences in platelet ag- gregation responses among the groups, as measured by impedance aggregometry of whole blood, to either 0.5 mM arachidonic acid or 5 4M adensoine diphosphate. To- tal cholesterol and LDL were elevated in plasma by the C and E diets in comparison to the N diet; the high- density lipoproteins were not different. Triglyceride levels in the E group were greater than in the N group. LDL was isolated by ultracentrifugation from C and CE diet animals, and thiobarbituric acid reactive substances (TBARS) were determined by fluorescence spectroscopy. TBARS were lower in the CE group. Dietary supplemen- tation of vitamin E did not alter platelet aggregation re- sponses but did reduce the oxidation of LDL in the hy- percholesterolemic animals. Effect of exercise and tamoxifen on induction of estro- gen-dependent and estrogen-independent rat mammary tumors. ROBERT DOEPKE* and DAVID MAGRANE, Department of Biological and Environmental Sciences, Morehead State University, Morehead, KY 40351. This research characterized the influence of moderate levels of exercise and the antiestrogen tamoxifen on in- duction and development of estrogen-dependent and es- trogen-independent rat mammary tumors. Fifty-day-old female Sprague-Dawley rats were given a 10 mg dose of the carcinogen 7,12-dimethylbenz(a)anthracene (DMBA) Abstracts, 1995 Annual Meeting 63 and then placed in one of four groups: I—Controls, II— Exercise, I1]—Tamoxifen, and IV—Tamoxifen + Exer- cise. Groups II and IV were given moderate treadmill ex- ercise (15 minutes/day; 5 days/week/12 weeks) at a belt speed of 20 meters/minute with a 1° incline. Groups III and IV were given subcutaneous injections of 100 wg of tamoxifen citrate (TAM) in oil 5 days/week for the 12 weeks. Results show that rats receiving TAM had signifi- cantly lower body weights compared to controls. Rats in the exercise group had 54% fewer total tumors than con- trols after 12 weeks; rats receiving TAM alone or TAM + exercise each had 92% fewer tumors. Rats with tumors were ovariectomized, and tumors that regressed by 20% after a 2-week period were judged estrogen dependent. Control rats showed 47% estrogen-dependent tumors; the exercise group had 80% estrogen-dependent tumors; and the TAM group and TAM + exercise were all estrogen independent. This research determined that both mod- erate exercise and tamoxifen showed a reduction in tu- morigenesis in the DMBA rat model. Renal response to dehydration in the larval bullfrog (Rana catesbeiana). ENDANG L. WIDIASTUTI* and JOHN J. JUST, School of Biological Sciences, University of Kentucky, Lexington, KY 40506. Bullfrog tadpoles can osmoregulate when placed in a wide range of medium osmolalities (7-200 mOsm/kg H,O). Part of physiological mechanisms involved in this hyperosmotic regulation is an alteration of kidney func- tion. About 125 tadpoles were cannulated; their urine was collected for ca. 24 hours. A decrease in urine formation rate (UFR) occurred during periods of dehydration. A group of 49 tadpoles maintained in artificial pond water (APW) at 7 mOsm/kg H,O had a UFR of 12.0 + 1.0 pl/ g/hr; a group of 43 tadpoles maintained in 100 mOsm/kg H,O of mannitol in APW had a UFR of 3.8 + 0.4 pl/g/ hr. Finally 39 animals maintained in 200 mOsm/kg H,O of mannitol in APW had a UFR of 2.1 + 0.3 wl/g/hr. The decrease in UFR was accompanied by an increase in urine concentration from 47 + 3 mOsm/kg H,O in normal APW, up to 62 + 4 mOsm/kg H,O in 100 mOsm/kg H,O of mannitol in APW, and as high as 120 + 6 mOsm/kg H,O in 200 mOsm/kg H,O of mannitol in APW. The ratio of urine osmotic pressure to plasma osmotic pressure (U/ P ratio) increased during dehydration, from 0.30 + 0.02 in the normal APW, to 0.40 + 0.04 in 100 mOsm/kg H,O of mannitol in APW, to as high as 0.57 + 0.03 in 200 mOsm/kg H,0 of mannitol in APW. Tadpoles were grouped into different Taylor-Kollros stages of develop- ment (