= ¥ 7 ' athe”! i abigns ms vee i + 4 Ke A SLR (NAGY eee Peder aes A i ih bie Si ‘hath , ST OPUS a UNTER : a Nieug si divdinad vibath sinc teed U tie PL Sk G ti Hits bei fi 43 ; va. 3 FULD 4 NG AGN cid eb ty i e : ‘ ‘ p g FECHA SGD ARAM HEI SG Sha SIA tf . - - SPM NEAL abel ches - ¢ RAR NG Apwipol ne Ud yas Fs ) Deis Tarai i ay 18 Spero ian FT pr tetieleh bin ; Fs ‘ 2 é 3 Dat vata reed ? res : $ 7 a. 3 3 “4 sRatb decd staily yen ies pak a be he tite ¥ j : ° So LC GE aT Sa yin et dct Pree e e) g WR HE 4) Fees jsealbuibw eens Get « i me Adevt ast bad ts td HM itt sae ¥ re Hi is t, ib * NIN IRS re - i080 ee o fa $ LN nae Hate} 2 i ote Bs hens Ih ie Las nats Havin V9 We B o i doin Gade th Waste He a Seta Do tah ov gis Bey pee yh aye coy AMEE Fapadeiianodaates et uviedie die Ovals Mitr daina fe: r z i ' f fe a 3 y si he é ea ‘ ASN AAC aT ah A bed whe ah ech win whe rate Ay da, dasa e Bails IS DRANG oid #4 «Mhaskali oi ural deed tat Teswetime gj i 4 ves Pty ade x : : : : a! Sohn iche fen Jens erie F ; " Bel ae on bet fala eb ae VE ae ewe rene ens Senate anew aie Te fieeee Teds PRES ER 2 ed ING iieg = SoM ated, Peo tee ae oF Thy : re peal Fic ¥ 7 P Sart - os Seth Ooo od heated = = ni ¥ it Hetine an ay AVM Bhatt 0 i a yk, Veter cae ek au [EONS nH x Oey | ta ase ie ie REN aut NR. ares or an i Py tee vt A with A i tN Ne, Ne) Hanh ' Ny rl eatin NE «tem Ny ( ac ae ite i, Kaa HRN eae) tH ‘ FAB, TRUE ea au iG HA Wee ie aN CASE i ea ih ey yh Dian Aue P Tse Meoailen! 7 Ueaie tt Pmt TAINAN ri N, Nat nen WAS 2) al Bs es] pemerhs eel 7 } ‘ MP REA Tee men PP PART CA un 7 Thi ha p Eva ALD a Ptr bei a Wai tind | i ; i Nain f , : i ‘ee * ei oneh Py ar 8 ; ; 5 mee Toi ne AVeths a] Ty i TRANSACTIONS OF THE © KENTUCKY ACADEMY OF SCIENCE Volume 58 Number 1 March 1997 _ Official Publication of the Academy The Kentucky Academy of Science Founded 8 May 1914 Governinc Boarp 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; (606) 572-6390 Editor, NEWSLETTER (ex officio): Maria K. Falbo-Kenkel, Department of Physics and Geology, Northern Kentucky University, Highland Heights, KY 41099 MEMBERS, GOVERNING BOARD Robert J. Barney 1999 J. G. Rodriguez 1998 Patricia K. Doolin 1996 AAAS/NAAS Representative David E. Hogan 1996 Wimberly C. Royster 1997 James F. Hopgood 1998 Robert H. Wombles 1998 Gerald L. DeMoss 1997 Vincent DiNoto 1996 Chair, KJAS COMMITTEE ON PUBLICATIONS Editor and John W. Thieret, Department of Biological Sciences, Northern Kentucky University, Chair: Highland Heights, KY 41099 Associate Editor: James O. Luken, 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. Creek, Department of Biological Sciences, Eastern Kentucky University, Richmond, KY 40475 Larry P. Elliott, Department of Biology, Western Kentucky University, Bowling Green, KY 42101 All manuscripts and correspondence concerning manuscripts should be addressed to the Editor. The TRANSACTIONS are indexed in BIOSIS and in State Academies of Science Abstracts. 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: $45.00 domestic; $50.00 foreign. Back issues are $30.00 per volume. The TRANSACTIONS are issued semiannually in March and September. Two numbers comprise a volume. Correspondence concerning memberships or subscriptions should be addressed to the Secretary. This paper meets the requirements of ANSI/NISO Z39.48-1992 (Permanence of Paper). 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 Bellarmine College Hazard Community College Berea College Paducah Community College Campbellsville College Somerset Community College Centre College Southeast Community College Cumberland College Associate Member Ashland Community College Maysville Community College Georgetown College Midway College Jefferson Community College | Owensboro Community College Kentucky State University Spalding University Kentucky Wesleyan College Thomas More College Transylvania University INDUSTRIAL AFFILIATES Associate Patron Ashland Oil, Inc. Member Corhart Refractories Corporation Hoechst Celanese Corporation MPD, Inc. United Catalyst, Inc. Westvaco Associate Member All-Rite Pest Control Lockheed Martin Wood Hudson Cancer Research Laboratory, Inc. nue Tiel Vert Le eae Obs (kit risReetny Ter kane Can hy we Gpe Janay TRANSACTIONS of the KENTUCKY Trans. Ky. Acad. Sci. 58(1):1-3. 1997. ACADEMY of SCIENCE March 1997 Volume 58 Number 1 Relationships Among Habitat, Cover, and Eastern Cottontails (Sylvilagus floridanus) in Kentucky William M. Giuliano! and Charles L. Elliott Department of Biological Sciences, Eastern Kentucky University, Richmond, KY 40475 and Jeffery D. Sole Kentucky Department of Fish and Wildlife Resources, Frankfort, KY 40601 ABSTRACT We examined relationships among habitat, cover, and eastern cottontails (Sylvilagus floridanus) on five wildlife management areas in Kentucky in 1989. The pellet plot technique was used to determine habitat- use patterns and preferences, and a vegetation profile board was used to determine foliage cover. Old fields and fencerows contained the greatest foliage cover and were preferred by cottontails year-round. Forested areas and agricultural fields were not high-use areas and were typically avoided. INTRODUCTION The eastern cottontail (Sylvilagus floridan- us), the most widely distributed species of cot- tontail, is one of the most sought after game animals in America (Chapman and Feldhamer 1982). In Kentucky, the cottontail provides recreation and sport for thousands of hunters each year. However, its numbers have been declining in the state over the last several years (Sole 1989). Changes in land use may be facilitating this decline (Bruna 1952). Intensive agricultural practices (cc over- grazed pastures, large monocultures) typically result in loss of valuable rabbit habitat such as fencerows and other shrubby, so-called waste areas (Bruna 1952). Without areas such as these, food and cover for rabbits may be lim- ' Current address: Department of Biological and Envi- ronmental Sciences, California University of Pennsylvania, California, PA 15419. iting their numbers (Barbour and Davis 1974). Succession of fields to forests may also reduce quality and quantity of rabbit habitat. Our ob- jective was to examine relationships among habitat, cover, and eastern cottontails to pro- vide information that may be key to conserv- ing cottontails in regions such as Kentucky with declining populations of these animals. METHODS We conducted our study on five wildlife management areas (WMAs) operated by the Kentucky Department of Fish and Wildlife Resources: Central Kentucky WMA (Madison County), Clay WMA (Nicholas County), Yel- lowbanks WMA (Breckinridge County), Tay- lorsville Lake WMA (Spencer County), and Lloyd WMA (Grant County). We classified ar- eas on each WMA into one of four habitat es: (1) fencerows: shrub- and tree-domi- nated (> 25% cover) strips of vegetation rang- ing from 1 m to 30 m wide; (2) forests: areas 2 Transactions of the Kentucky Academy of Science 58(1) Table 1. Foliage cover and habitat use by eastern cottontails on five wildlife management areas in Kentucky, 1989. Foliage cover (%)" Cottontail use (%) Mar May Jul Sep Nov 42A 4A 43A 50A 42A 538A 44A SDA 49A 56A 5B 2B 2B 1B 2B OB OB OB OB OB Habitat type 76A 46A Old field TTA 52B 49A Agricultural field = — 4B Forest 32B 30C 1B ‘Foliage cover determined August 1989. ' Means in a column followed by the same letter are similar (P > 0.05). of closed hardwood-dominated forest, with tree cover >25%; (3) old fields: habitat rang- ing from pastures of tall grass to older succes- sional stages containing abundant woody shrubs and small trees, with tree cover =25%; and (4) agricultural fields: primarily corn fields, with some winter wheat planted in fall and winter. Two areas of each habitat type per WMA were randomly selected for study. Habitat use by cottontails was determined using the pellet plot technique (Pietz and Tester 1983). We es- tablished and marked with wooden stakes 25 plots, each 1 m?. Plots were 10 m apart along randomly located transects running through forest, old field, and agricultural field sample areas. In fencerows, transects were located along the midline (total = 50 plots/habitat type/ WMA). We counted and removed fecal pellets from plots every 4 to 8 weeks between 20 Nov 1988 and 20 Dec 1989. We tested the null hy- potheses that cottontail use did not differ among habitat types, months, and WMAs with a factorial ANOVA followed by a Fisher's LSD test (SYSTAT 1992). We examined habitat pref- erences using chi-square tests (Neu, Byers, and Peek 1974; SYSTAT 1992) by testing the null hypothesis that cottontail use of habitats was random. All four habitat types were sampled with the same intensity (ie., 50 plots, each 1 m?/WMA). If use was random, each habitat type should have received 25% of the total use. We considered cottontail preference of a hab- itat type if use was greater than expected based on a random habitat-use pattern, and avoidance of a habitat type if use was less than expected based on a random habitat use pattern. A vegetation profile board was used to de- termine horizontal foliage cover in each hab- itat type in August 1989 (Nudds 1977). We estimated the percent of the profile board ob- structed by vegetation when viewed from a distance of 10 m, every 25 m along each 250 m transect in each habitat type. Foliage cover was compared among habitat types and WMaAs using a factorial ANOVA followed by Fisher's LSD test (SYSTAT 1992). RESULTS Cottontail use (Table 1) differed among habitat types (P<0.05) but not among months or WMAs (P>0.05). Habitat preferences did not differ by month (P>0.05). Fencerow and old-field habitats were preferred (P<0.05), and forests and agricultural fields were avoid- ed (P<0.05). Foliage cover (Table 1) differed by habitat type (P<0.01) but not WMA (P>0.05). Fo- liage cover estimates of agricultural fields were not used because of high variability in cover due to differences in crop rotations and prob- lems associated with frequent disturbance (i.e., harvest and planting activities). DISCUSSION The cover that habitats provide is the most influential feature determining use of an area by rabbits (Anderson and Pelton 1976; Swihart and Yahner 1982). Our findings agree with those of Barbour and Davis (1974) and suggest that fencerow and old-field habitats consistently received the most use and were preferred be- cause they had greater foliage cover than forest habitats, at least in summer. The added cover in old fields is important for escape and nesting (Bruna 1952). The additional cover provided by fencerows may have been important as a source of protection, escape routes, and travel corri- dors (Edwards et al. 1981). In fall and winter, old fields usually lose much of their value to cottontails, both as a source of food and cover, due to desiccation of herbaceous vegetation and loss of foliage from woody vegetation. Because of this, rab- Habitat, Cover, and Cottontails—Giuliano and Elliott bits typically move to forested areas and other areas with abundant woody vegetation that provides adequate cover (Barbour and Davis 1974; Lewis 1972). During our study, howev- er, cottontails continued to prefer old fields and to avoid forested areas even in fall and winter. We believe this may have been due to an adequate amount of cover being provided by old fields, even in fall and winter. In addi- tion, the winter of 1988-1989 was mild, with below average snowfall (USDC-NOAA 1988- 1989). Therefore, rabbits may not have need- ed to move to areas containing more woody vegetation in search of food and cover (Han- sen, Bailey, and Siglin 1969). We suggest that limited use and apparent avoidance of agricultural fields was probably due to the continuous disturbance associated with agricultural practices (Friley 1955). Ed- wards et al. (1981) reported similar patterns of use and avoidance of agricultural fields by cottontails in Illinois. We have shown that fencerows and old fields are preferred, year-round cottontail hab- itats; the creation and maintenance of these areas should be a high management objective. Forests and agricultural fields are not high-use areas but may be seasonally important as sources of food and cover. We suggest that management activities include (1) maintaining and planting of shrubs and trees along fences and field edges to enhance fencerow habitat, (2) disking, mowing, and burning of fields to prevent succession to forest, and (3) coordi- nating agricultural practices to minimize dis- turbance of fields and provide additional sources of food. ACKNOWLEDGMENTS This study was funded by Pittman-Robert- son Federal Aid in Wildlife Restoration mon- ies in cooperation with the Kentucky Depart- ment of Fish and Wildlife Resources, P-R Pro- ject W-45-21, and the Wildlife Program, De- partment of Biological Sciences, Eastern Kentucky University. ioe) LITERATURE CITED Anderson, B.F., and M.R. Pelton. 1976. Movements, home range, and cover use: factors affecting the sus- ceptibility of cottontails to hunting. Proc. Southeast. As- soc. Game and Fish Commissioners 30:525-535. Barbour, R.W., and W.H. Davis. 1974. Mammals of Ken- tucky. Univ. Press of Kentucky, Lexington. Bruna, J.F. 1952. Kentucky rabbit investigations. Ken- tucky Fish and Wildlife Dep., Fed. Aid. Proj. 26-R., Frankfort. Chapman, J.A., and G.A. Feldhamer. 1982. Wild mam- mals of North America. Johns Hopkins Univ. Press, Bal- timore, MD. Edwards, W.R., S.P. Havera, R.F. Labisky, J.A. Ellis, and R.E. Warner. 1981. The abundance of cottontails in relation to agricultural land use in Illinois (U.S.A.) 1956-1978, with comments on mechanism of regula- tion. Proc. World Lagomorph Conf. 1:761-789. Friley, C.E. 1955. A study of cottontail habitat prefer- ences on a southern Michigan farming area. Fed. Aid Proj. W-48-R. Hansen, J.C., J.A. Bailey, and R.J. Siglin. 1969. Activity and use of habitat by radio-tagged cottontails during winter. Trans. Ill. Acad. Sci. 62:294—302. Lewis, H.S. 1972. Cottontail rabbit home range, habitat use, and mortality in southern Wisconsin. M.S. thesis, Univ. Wisconsin, Madison. Neu, C.W., C.R. Byers, and J.M. Peek. 1974. A technique for analysis of utilization-availability data. J. Wildlife Managem. 38:541—545. Nudds, T.D. 1977. Quantifying the vegetative structure of wildlife cover. Wildlife Soc. Bull. 5:113-117. Pietz, P.J., and J.R. Tester. 1983. Habitat selection by snowshoe hares in north central Minnesota. J. Wildlife Managem. 47:686-696. Sole, J.D. 1989. Quail and rabbit roadside survey. P-R Proj. W-45-21. Ky. Dep. Fish and Wildlife Resources, Frankfort. Swihart, R.K., and R.H. Yahner. 1982. Habitat features influencing use of farmstead shelterbelts by the eastern cottontail (Sylvilagus floridanus). Am. Mid]. Naturalist 107:411-414. SYSTAT. 1992. SYSTAT for Windows: statistics, Version 5 ed. SYSTAT, Inc. Evanston, IL. [USDC-NOAA] U.S. Department of Commerce-National Oceanic and Atmospheric Administration. 1988-1989. Climatological data: Kentucky. Vol. 83, No. 13; Vol. 84, Nos. 1-3. ISSN 0145-0433. National Environmental Satellite, Data and Information Service, National Cli- matic Data Center, Asheville, NC. Trans. Ky. Acad. Sci. 58(1):4-8. 1997. Separation of Spawning Habitat in the Sympatric Snubnose Darters Etheostoma flavum and E. simoterum (Teleostei, Percidae) Jean C. Porterfield Fish Division, Museum of Zoology, Ann Arbor, MI 48109! ABSTRACT Individuals of two snubnose darter species, Etheostoma flavum and E. simoterum, were observed in Whites Creek, Tennessee, in April 1995 and April 1996. Both species were present and spawning in 1995, but in 1996 the stream had experienced increased siltation and algal growth, and no E. flavum individuals were seen. Spawning behaviors of both species were similar to those known for other snubnose darters: E. flavum and E. simoterum attached their eggs to rock surfaces, and the male followed the female as she searched for appropriate substrate. Behavioral observations in 1995 suggested that these two species utilized different sites for spawning, and that interspecific aggression was not the mechanism by which habitat isolation was obtained. Since most closely related darters have allopatric distributions, studying instances of sympatry such as this may contribute to understanding of distribution patterns of darters as a group. INTRODUCTION Theoretical works on species concepts and speciation processes traditionally cite repro- ductive behavior as a significant isolating mechanism between sympatric species (Mayr 1963; Paterson 1993). Choice of spawning site is one aspect of reproductive behavior that has been proposed as an isolating mechanism in some fishes (lamprologine cichlids, Ribbink et al. 1983; Lake Baikal sculpins, Smith and Todd 1984), and it may be important in some spe- cies of darters. Darters are one of the most speciose groups of freshwater fishes in North America, and it is common to find more than 10 species in a single stream. Although most closely related darters are allopatric, some do share overlapping distributions. Several stud- ies have demonstrated the significance of mi- crohabitat segregation in competition among sympatric darters, although reproductive be- haviors were not specifically addressed (Greenburg 1988; Kessler and Thorp 1993). The snubnose darters, members of the sub- genus Nanostoma (sensu Page 1981, but see Bailey and Etnier 1988) in the genus Etheos- toma, are comprised of 23 described species, most of them restricted to the southeastern United States (Page and Burr 1991) and most distributed allopatrically. Etheostoma simoter- um, the snubnose darter, has a wide range throughout the Tennessee and Cumberland ' Present address: Illinois Natural History Survey, 607 East Peabody Drive, Champaign, IL 61820. river systems where it is often sympatric with two other species of snubnose darters: Eth- eostoma duryi, the black darter, and E. fla- vum, the saffron darter. Etheostoma flavum is sympatric with E. simoterum throughout much of its range in the lower Cumberland and Duck river systems and in some tributar- ies of the lower Tennessee River (Etnier and Stamnes 1993). Spawning behavior has been described from aquarium observations for E. simoterum (Page and Mayden 1981) and from both aquarium and field observations for E. flacum (Keevin, Page, and Johnston 1989); both species are classified as egg attachers (Page 1985). Descriptions of both species can be found in Etnier and Starnes (1993). Here I describe the spawning behavior of E. simo- terum under natural conditions, present the results of observations on spawning habitat separation between E. simoterum and E. fla- vum in Whites Creek, Tennessee, and discuss the implications for reproductive isolation be- tween the two species. MATERIALS AND METHODS I observed spawning E. simoterum and E. flavum while snorkeling in Whites Creek at Hwy 41A bridge, Davidson County, Tennes- see, on 8, 9, and 16 April 1995, and on 11 April 1996. Observations were made in all available habitat types including silted bedrock pools, gravel/cobble runs and riffles, and al- gae-covered boulders (rip rap) piled at stream edges; depths ranged from 30 cm to 1 m. In 1995, behaviors of both species were filmed Spawning Habitat in Snubnose Darters—Porterfield 5 on 9 and 16 April using a Sony CCD-FX310 8-mm video camera in a Sony MPK-TRA2 un- derwater housing. Observations were record- ed at the spawning sites and later when I viewed the video tapes. I observed E. simo- terum at the bridge for a total of 4 hours on 2 separate days, during which time four spawning pairs were followed; additionally, 2 hours of observations including one spawning pair were made over the 2 days in a bedrock pool at a site about 300 m upstream. Three hours of observations on six individuals of E. flavum were made on the same 2 days at the bridge. I observed the darters from late morn- ing to mid-afternoon each day. Water temper- ature during the observations remained around 21° C. On 11 April 1996 fish were observed in the late morning and early afternoon. The water temperature was 11° C. No darters were filmed, but observations on species present, behaviors of these species, and habitat condi- tion were noted. RESULTS Although courtship behavior prior to pair- formation was not observed in either E. sim- oterum or E. flavum, the spawning behavior after pair-formation was observed to be iden- tical in both species. The male closely fol- lowed the female as she swam over the sub- strate searching for egg deposition sites. He stayed above and a little behind her with me- dian fins semi-erect; in some cases he left to chase an approaching male. When the female found a potential site for egg-deposition, she visually examined the site, as her body formed a 30-45 degree angle with the substrate sur- face. She then made one to four sharp jabs at the spot with her snout as the male moved to position himself above her. The head jabs ob- served were similar to those described for E. davisoni by Bart (1992) and for E. zonale by Walters (1994) and were a consistent indicator of oviposition, with 100% of all spawns in both species preceded by at least one head jab. Im- mediately following the head jabs, the female dragged her body over the chosen site as the male hovered closely above her. When the female’s genital papilla reached the site, she pressed her papilla tightly against the rock and oviposited, while the male arched his body against her, pressed his caudal peduncle to- wards the rock, and vibrated, releasing sperm. This spawning behavior is identical not only between E. simoterum and E. flavum but also among the other species in the subgenus Na- nostoma that I have observed spawning in the field: E. barrenense, E. duryi, and E. rafines- uei. In 1995, E. simoterum was the most com- mon fish near the Hwy 41A bridge and the most common darter at the riffle/pool habitat upstream of the bridge. Many individuals were seen in the large cobble raceway near the cen- ter of the stream; although individuals of both sexes were observed here, there were more females, and no E. simoterum were observed forming pairs or spawning in this area. Fewer E. simoterum were present among the large algae-covered boulders piled near the bridge on the north side of the stream; however, there was a greater proportion of males seen here, and the only spawning pairs of E. sim- oterum at the bridge site were located in these piles. I observed pairs attaching eggs to the boulders, and spawning positions ranged from slightly angled near the tops of boulders, to near vertical positions on the sides of boul- ders, to upside-down under an overhanging boulder. No other darter species were ob- served in this large-boulder area, although some E. caeruleum were seen at its periphery. In 1995, E. simoterum was common at the upstream site near the sides of the stream in areas of algae-covered flat bedrock with some cracks and shelves. There were many individ- uals in the area, but only one spawning pair was observed. The pair was observed on a bedrock shelf spawning in an algae-covered crevice in a horizontal position. At both sites, individuals spawned on surfaces covered with algae; out of the total of nine spawns observed, eight occurred on large algae-covered boul- ders or stretches of bedrock in a flowing pool environment at the side of the stream. The other spawn took place on a piece of cobble also covered with algae but located closer to the raceway in faster water. The six individuals of E. flavum observed in 1995 were found only in the cobble raceway above the Hwy 41A bridge. This raceway av- eraged 60 cm deep; the substrate consisted of gravel and cobble. Two spawning pairs were observed, and two other individuals were seen. In all four spawning bouts the females 6 Transactions of the Kentucky Academy of Science 58(1) chose to attach eggs to pieces of clean rock in the fastest current of the raceway. Three spawns took place on pieces of cobble, while the fourth took place on a side of a rock ledge. Spawning positions were most often at about a 45 degree angle to the substrate surface as females oviposited on the sides of the rocks. Etheostoma simoterum and E. caeruleum were the two most common fishes in the raceway, but I did not observe individuals of either spe- cies spawning there. There were no interspecific interactions be- tween spawning pairs of E. simoterum and E. flavum individuals in Whites Creek, but this observation is affected by the demography in the stream; E. flavum was not present in the boulders where E. simoterum spawned. How- ever, spawning E. flavum pairs often encoun- tered E. simoterum individuals. Encounters are defined as fishes coming within one body length of each other. Of 11 of the interspecific encounters documented for the E. flavum pairs, two were with E. simoterum males. In one instance the E. flavum individuals ignored an E. simoterum male who swam between them; in the other instance the E. flavum male swam ahead of the female to approach an E. simoterum male who seemed to be feeding in their path. None of the interspecific encoun- ters between single individuals (i.e., not spawning pairs) involved aggression. Of two encounters between E. flavum pairs and con- specific males, both involved aggressive inter- actions; for E. simoterum all seven intraspe- cific encounters involved aggression. Observations in Apri! 1996 revealed that the habitat in Whites Creek was different from the habitat observed a year earlier. The clean cobble in the faster run where E. flavum was seen in 1995 was covered with algae, and the whole area appeared to have experienced more siltation. Both male and female E. sim- oterum were common in the cobble runs, but no E. flavum individuals were seen. Large E. simoterum males were still common among the algae-covered boulders at the side of the stream, and some E. simoterum females were present as well. No spawns were observed, but three pairs of E. simoterum were seen: one among the boulders and two in the cobble area at the head of the raceway. Fish stream associates in 1995 at the bridge site were Campostoma anomalum, Etheosto- ma caeruleum, E. crossopterum, E. flabellare, E. smithi, Lepomis cyanellus, L. megalotis, Luxilus chrysocephalus, Micropterus punctu- latus, and Percina caprodes. Associates in 1995 at the upstream site were Campostoma an- omalum, Etheostoma blennioides, Lepomis megalotis, Luxilus chrysocephalus, Lythrurus ardens, Micropterus punctulatus, and Pime- phales notatus. Observed fish associates in 1996 at the bridge site were Campostoma an- omalum, Catostomus commersoni, Etheostoma caeruleum, E. flabellare, Lepomis cyanellus, L. megalotis, Luxilus chrysocephalus, and Mi- cropterus punctulatus. DISCUSSION The spawning behaviors of E. flavum and E. simoterum observed in Whites Creek are nearly identical to previously published de- scriptions for these two species (Keevin, Page, and Johnston 1989; Page and Mayden 1981). I found the angle of the spawning position for E. flavum to be around 45 degrees; Keevin, Page, and Johnston (1989) reported a near vertical spawning position for aquarium-held fish. These differences may reflect the cobble habitat in which E. flavum was found in Whites Creek as opposed to the large flat rocks preferred by aquarium-held Warren Fork fish (Keevin, Page, and Johnston 1989). Males of neither species defended stationary territories; instead, females would roam a large area of substrate searching for oviposi- tion sites, and males defended the females with which they were paired. These observa- tions are similar to those reported in studies of the spawning behaviors of other snubnose darters (Carey and Burr 1989; Keevin, Page, and Johnston 1989; Page and Mayden 1981; Stiles 1974). Observations of E. flavum and E. simoterum in Whites Creek in 1995 suggest that in this stream these two snubnose darters utilize dif- ferent spawning sites in different areas of the stream. Previous studies have shown that un- der natural conditions, both species preferred flowing-pools with a variety of substrates; in aquaria they preferred to deposit eggs on large stones (Keevin, Page, and Johnston 1989; Page and Mayden 1981). However, Keevin, Page, and Johnston (1989) added that when large rocks are not available, E. flavum individuals spawn on a cobble substrate. I found E. fla- Spawning Habitat in Snubnose Darters—Porterfield i vum to be utilizing the clean cobble in the swifter runs, while E. simoterum spawning pairs were found among the large, algae-cov- ered boulders in slower current at the side of the stream. These data support the hypothesis that E. simoterum individuals in Whites Creek occupy the boulder pool habitat, making it un- available to E. flavum individuals. Etnier and Stames (1993) stated that E. simoterum indi- viduals in sympatry with E. flavum are more often found in the swifter riffles. My obser- vations in Whites Creek suggest that while many E. simoterum were seen in the faster raceway, this is not the area utilized for spawn- ing. Spawning pairs of E. simoterum were found only in the boulder pools, suggesting that this species may use different stream hab- itats for spawning than for other activities such as feeding. In addition, the presence of E. sim- oterum in the faster water is not a result of interactions with E. flavum, as individuals of the latter species were not seen in the pools. Behavioral observations show that members of the two snubnose darter species rarely in- teract aggressively with heterospecific individ- uals, while intraspecific interactions among males usually involve aggression. The greater number of interspecific encounters of E. fla- vum pairs with E. simoterum females (nine, versus two with E. simoterum males) reflects the demography of the E. simoterum popula- tion in Whites Creek; females were concen- trated in the faster current of the raceway where the E. flavum individuals were spawn- ing. Similarly, E. simoterum spawning pairs had no interspecific encounters because no E. flavum individuals were present in the boulder areas where they spawned. In part because of this relationship to demography, the rarity of direct aggression observed between species does not reject the hypothesis that competi- tion for spawning sites causes E. flavum to spawn in the cobble raceway. However, the encounter data do suggest that agonistic be- havior is not the means by which habitat iso- lation is maintained. Other possible mecha- nisms for maintenance of the habitat segre- gation observed should be considered, such as more subtle behavioral cues or recognition of male color patterns. In 1995 there were low numbers of E. fla- vum in this section of Whites Creek. This de- mographic factor may affect and/or be affect- ed by the patterns of habitat use seen in this species as well as in E. simoterum. The rarity of E. flavum in this stream increases the like- lihood that a given observation of spawning habitat in this species is due to chance and not to substrate choice. However, the spawning substrate used by this species may affect its relative success in this stream if, for example, there is differential success of eggs in the dif- ferent habitats. In 1996, E. flavum individuals were not observed in Whites Creek at Hwy 41A, and there were fewer species of fishes in general. These observations might be ex- plained by the difference in water tempera- ture (21° C in 1995 versus 11° C in 1996), but E. simoterum was observed forming pairs in the 11° C water in 1996, so presumably the water was not too cold for darters to be pres- ent and active. Or the absence of E. flavum might be explained by the increased levels of siltation and algal growth since April 1995; the species spawned on clean cobble in 1995, sub- strate, which was no longer available in 1996. The absence of E. flavum might also be a re- sult of interactions with E. simoterum; greater numbers of E. simoterum competing for food and lower egg survivorship for E. flavum in the faster water are possibilities. At this point, hypotheses concerning which factors might be responsible for the absence of E. flavum are speculative. Etheostoma simoterum is also sympatric with E. duryi in the middle Tennessee River system; these two species may also exhibit spawning habitat segregation when sympatric (pers. obs.). Snorkeling observations in April 1995 at Gray Branch (Cypress Creek system, Alabama) revealed that E. duryi attached eggs to clean cobble and bedrock at the head of an island, while most E. simoterum pairs were spawning on cobble in the edge of a run on one side of the island. These observations are consistent with those reported by Etnier and Starnes (1993). Etheostoma flavum and E. duryi are occasionally sympatric, particularly in the upper Duck River system as discussed by Etnier and Bailey (1989), but I have not made snorkeling observations at any of these localities. In most cases, the species of darters found in a given stream are not closely related (i.e., in the same subgenus) and do not exhibit sim- ilar reproductive habits such as choice of mi- 8 Transactions of the Kentucky Academy of Science 58(1) crohabitat or spawning substrate. The result- ing pattern is that the darter species in most streams coexist without interspecific competi- tion for spawning sites. Field data on E. sim- oterum and E. flavum in Whites Creek based on limited observations made during one spawning season suggest that there is some ev- idence for competition for spawning sites be- tween closely related sympatric darters. If cor- roborated with experimental data, these re- sults could be significant to studies of the re- productive ecology and distributional history of snubnose and other darter species. For ex- ample, if competition is found to be a signifi- cant factor in the existence and/or persistence of darter species in a given area today, then its role in the historical distribution of darters could be explored. ACKNOWLEDGMENTS This research was supported by the Univer- sity of Michigan Department of Biology, the University of Michigan Museum of Zoology, the University of Illinois Graduate College, and the Illinois Natural History Survey. I thank W.E. Browne, P.A. Ceas and A.L. Utz for field help; A.K. Peters and G.R. Smith for the use of video equipment; and P.A. Ceas and L.M. Page for helpful comments on the manu- script. LITERATURE CITED Bailey, R. M., and D. A. Etnier. 1988. Comments on the subgenera of darters (Percidae) with descriptions of two new species of Etheostoma (Ulocentra) from southeast- ern United States. Misc. Publ. Mus. Zool. Univ. Mich. 175:1-48. Bart, H. L., Jr. 1992. Spawning behavior of Etheostoma davisoni Hay. Copeia 1992:537-539. Camey, D. A., and B. M. Burr. 1989. Life histories of the bandfin darter, Etheostoma zonistium, and the fi- rebelly darter, Etheostoma pyrrhogaster, in western Kentucky. Ill. Nat. Hist. Surv. Biol. Notes 134:1-16. Etnier, D. A., and R. M. Bailey. 1989. Etheostoma (UI- ocentra) flavum, a new darter from the Tennessee and Cumberland river drainages. Occas. Pap. Mus. Zool. Univ. Mich. 717:1-24. Etnier, D. A., and W. C. Starnes. 1993. The fishes of Tennessee. University of Tennessee Press, Knoxville. Greenberg, L. A. 1988. Interactive segregation between the stream fishes Etheostoma simoterum and E. rufili- neatum. Oikos 51:193—202. Keevin, T. M., L. M. Page, and C. E. Johnston. 1989. The spawning behavior of the saffron darter (Etheos- toma flavum). Trans. Ky. Acad. Sci. 50:55-58. Kessler, R. K., and J. H. Thorp. 1993. Microhabitat seg- regation of the threatened spotted darter (Etheostoma maculatum) and closely related orangefin darter (E. bel- lum). Canad. J. Fish. Aquatic Sci. 50:1084-1091. Mayr, E. 1963. Animal species and evolution. Harvard University Press, Cambridge. Page, L. M. 1981. The genera and subgenera of darters (Percidae, Etheostomatini). Occas. Pap. Mus. Nat. Hist. Univ. Kans. 90:1-69. Page, L. M. 1985. Evolution of reproductive behaviors in percid fishes. Bull. Ill. Nat. Hist. Surv. 33:275-295. Page, L. M., and B. M. Burr. 1991. A field guide to fresh- water fishes of North America north of Mexico. Hough- ton Mifflin, Boston. Page, L. M., and R. L. Mayden. 1981. The life history of the Tennessee snubnose darter, Etheostoma simoterum, in Brush Creek, Tennessee. Ill. Nat. Hist. Surv. Biol. Notes 117:1-11. Paterson, H. E. H. 1993. Evolution and the recognition concept of species. The Johns Hopkins University Press, Baltimore. Ribbink, A. J., B. A. Marsh, A. C. Marsh, A. C. Ribbink, and B. J. Sharp. 1983. A preliminary survey of the cich- lid fishes of rocky habitats in Lake Malawi. South Afr. J. Zool. 18:149-310. Smith, G. R., and T. N. Todd. 1984. Evolution of species flocks of fishes in north temperate lakes. Pages 45-69 in A.A. Echelle and I. Kornfield (eds). Evolution of fish species flocks. University of Maine at Orono Press, Orono. Stiles, R. A. 1974. The reproductive behavior of the Green and Barren River Ulocentra (Osteichthyes: Per- cidae: Etheostoma). Assoc. Southeast. Biol. Bull. 21:86- 87. Walters, J. P. 1994. Spawning behavior of Etheostoma zonale (Pisces: Percidae). Copeia 1994:818-821. Trans. Ky. Acad. Sci. 58(1):9-19. 1997. Scientists of Kentucky Common Names of Vascular Plants Reported by C.S. Rafinesque in an 1819 Descriptive Outline of Four Vegetation Regions of Kentucky Ronald L. Stuckey Herbarium in the Museum of Biological Diversity, Department of Plant Biology, College of Biological Sciences, The Ohio State University, 1315 Kinnear Road, Columbus, Ohio 43212 and James S. Pringle Royal Botanical Gardens, Box 399, Hamilton, Ontario, Canada L8N 3H8 In the flora of North America, knowledge is scarce on the sources and origins of the com- mon names applied to vascular plants. In bo- tanical literature, however, a large number of studies have been published with the purpose of providing common names of plants for ei- ther general or specific regions of the United States. McAtee (1913-1933), in a series of five papers published over a 20-year period, listed 410 common names with the geographic areas in which they were used, and 60 sources of information. His list, consisting mostly of plants of aquatic and marsh habitats, is of names learned during field work for the Bio- logical Survey in the United States Depart- ment of Agriculture. McAtee’s list, containing an index to the plant names in each of his five papers, provides an excellent working base for obtaining local common names from literature sources published during the first third of the 20th century or earlier. Constantine Samuel Rafinesque (1783- 1840) (Figure 1), the controversial, eccentric, sometimes considered erratic European bota- nist who traveled in eastern North America during the first third of the 19th century, pub- lished articles and books totaling over 900 ti- tles in botany. Many of these publications were on taxonomic botany, including cata- logues of floras (Rafinesque 1836, 1836-1838, 1840), geographical floras (Rafinesque 1817b, 1817c, 1819a, 1824), medical floras (Rafin- esque 1828, 1830), and studies of genera (Raf- inesque 1811, 1820). In his Medical Flora, Raf- inesque (1828, 1830) provided an English, French, and German name, one or more of- ficinal names (i.e., names used in pharmacy), and one to several “vulgar names” for each species, the last being additional names in En- glish used by members of the public but not preferred for scholarly communication. Oth- erwise, in these well-known publications, he did not provide common names for the plants listed or described, many of which were des- ignated by him as new taxa. In a few of his papers among this vast output of botanical in- formation, he reported the common names of vascular plants. Rafinesque traveled and explored for plants in Kentucky during the years 1818 to 1826 and was Professor of Botany and Natural History at Transylvania University in Lexington from 1819 to 1825. During this period he wrote a short article (Rafinesque 1819c; Figure 2) con- sisting of a descriptive outline of the vascular- plant vegetation in four major regions of the state. This paper was published in 1819 in the second number of the first volume of The Western Review and Miscellaneous Magazine, edited by William Gibbs Hunt (Rafinesque 1819c). In addition to providing general re- marks, Rafinesque characterized each of the four botanical regions as to geographical lo- cation, general topography, kinds of bedrock, and unique features of the vegetation. He also provided a short list of vascular plants that he considered somewhat specific or “peculiar” to each region. Along with each binomial scien- tific plant name, he added the common name, or in his terminology, the “vulgar” name, not- ing, however, that “the vulgar names of the plants .. . cannot claim to be generally under- stood even in [Kentucky], many being merely 10 Transactions of the Kentucky Academy of Science 58(1) | | 4M ~~ Figure 1. Portrait of C.S. Rafinesque by American artist William Birch (1755-1834). After a long succession of private owners, this enamel miniature (2 4” diameter) was purchased by Transylvania University, Lexington, Kentucky, in 1938. Photo courtesy of Transylvania University Special Collections. local or personal. The botanical names are alone to be relied on.” Two years earlier, in his review of the first edition of Amos Eaton’s Manual of Botany for the Northern States . . . [1817], Rafinesque (1817a) supported the idea of providing the vulgar names in works on local taxonomic bot- any, and he noted that their inclusion in Ea- ton’s Manual was “valuable.” He commented that most of them had been taken by Eaton from other cited published sources, rather than being “taken directly from the vulgar,” and wrote further that “vulgar names are at all times a valuable appendage to classical syn- onymy, and indispensable in local botanical writings.” Rafinesque probably prepared the article on vegetation of Kentucky for a popular audi- ence, which would explain his use of the com- mon names of plants, but he reminded his readers that the botanical names were the in- tended authentic ones to be used for scientific accuracy. Botanists always have and, it is ex- pected, always will recognize that the botani- cal names should be used in authoritative flo- ristic and vegetational studies. Botanists, how- ever, must also communicate with and make their studies useful to those who work in other fields of study but who are not familiar with Latin binomials. Among these fields are geol- ogy, zoology, wildlife and fisheries studies, nat- ural resources management, endangered spe- cies preservation, conservation organizations, environmental law, ecological consultation, and other groups with environmental con- cems. The use of common names of plants is of more importance now than it was 25 or 50 Rafinesque Common Names—Stuckey and Pringle 11 MISCELLANY, =; —=+ BOTANY OF KENTUCKY. ‘On its principal features, by C. S. Rarinesque, Professor of Botany and Natural History in Transylvania Universi- ty. THE state of Kentucky being situated in the-centre of the western country, has a flora similar to the generality of the western states, and participating in their peculiar features, while’ if offers in itself a complete specimen of the western botany. The peculiarities of this botany consist principally in the total want of the maritime and mountain regions, which form sach remarkable sections in the local floras of the Atlantic states, and abound with plants peculiar’ to themselves. Another striking feature in the vegetation of Kentucky and the western states is the propensity which many plants and trees exhibit of growing in a-social state, to the almost total exclusion of every other. There are many plants which grow crowded together, allover the United States; such for instance as the grasses, ferns, the ‘Comptonia, the Studfonia, &c. but they al- low many other plants to grow with them; while, in-the western country, many extensive spaces of ground are covered with one or a few crow: ‘ed species, to the exclusion of many others, which are found in their company elsewhere. The plants which may be quoted as a striking 3 instance of this singular fact are not few, among which I shall select the following: Vernonia frrealta, Tron Weed,. Bapttisia cerulea, - Blue. Wild Indigo, . Cacalia reniformis,. ~ Kidney Weed, Stedeoma pulegivides, ’Penny-aoyal, Chenopodium SU Worm ‘Weed, Elephantofius scaber, ‘Elephant’ s Fo ts ‘ Figure 2. First page of C.S. Rafinesque’s paper on “Bot- any of Kentucky[:] on its principal features,” published in 1819 in volume 1(2) of The Western Review and Miscellaneous Magazine. which was years ago; it will become more and more prev- alent and necessary as time progresses. With increasing frequency, floristic manuals, such as that of Gleason and Cronquist (1991) and the ongoing Flora of North America North of Mex- ico, are providing vernacular names for all or most of the species described therein. The use of common names will demand more accuracy and stability to the extent that nomenclatural guidelines may need to be established in the future. These guidelines may develop in a manner similar to those parameters provided in the International Code of Botanical Nomen- clature. The correct interpretation of earlier papers in which common names were used re- quires that the authors’ applications of such names be accurately determined. Some names may have been applied in senses different from more recent usage, and some may have fallen into disuse. Efforts to standardize com- mon nomenclature require the selection of ap- propriate and otherwise acceptable names that will not perpetuate or introduce confusion. At- taining these objectives will require consider- ation of historic and regional applications of vernacular plant names. For these reasons we bring to the attention of the scientific public the common plant names in Rafinesque’s early 19th century publication on the botany of Kentucky. Rafinesque’s paper on the botany of Ken- tucky is transcribed below. His lists of plant species are annotated, as indicated in the fol- lowing paragraphs, to permit their interpreta- tion using current botanical and common names. Separate indexes to botanical and com- mon names are provided in Appendix 1. Rafinesque—owing, he said in the errata, to his absence from town—had no opportunity for proofreading until after the paper had gone to press. Consequently, the original lists of plant names are rife with typographical er- rors, even among the common names. Rafin- esque (1819d) published corrections of a num- ber of these errors in a later issue of the same journal, on page 128. Several others, however, escaped his notice. “Tris” for “Iris,” and “Pin week” for “Pin weed,” for example, are obvi- ous typographical errors. In some cases, however, minor alterations presumably do represent Rafinesque’s intent. From some of his other publications, it is ev- ident that he preferred a simplified spelling such as “cerulea” to the original “caerulea” or “coerulea” and “catesbei” for “catesbaei,” and that he frequently made such arbitrary abridg- ments. Such changes, whether required or proscribed by present rules of nomenclature, are not regarded as new names, and they re- main attributed to the original authors. Be- cause of the circumstances discussed above, it appeared best to retain Rafinesque’s original format in the lists, which omitted the attri- bution of the authorship of the botanical names. As the lists appear in the present pa- per, the botanical names used by Rafinesque are in italic type; the common names, in ro- man type. (At the left margin we have num- bered, in parentheses, each species for index- ing purposes.) Indented and in square brack- ets and smaller type below the names we have added, first, the corrected orthography of the 12 Transactions of the Kentucky Academy of Science 58(1) botanical name used by Rafinesque, in italics if it is not the currently accepted botanical name, with authorship; second, where appro- priate and in roman type, the currently ac- cepted botanical name; and third, a common name or names in current use. When the bo- tanical name used (or intended) by Rafinesque remains in use for the species, this name ap- pears in roman type, with the orthography cor- rected if necessary and the authorship indi- cated. Curly brackets have been used to in- dicate the corrections made by Rafinesque (1819d) in the errata. In Appendix 2, notes, mostly on nomenclature, have been added for 14 taxa. The moder common names we include have generally been obtained from Fernald (1950), Gleason and Cronquist (1991), and the popular field guides by Peterson and Mc- Kenny (1968) and Newcomb (1977). The orig- inal Illustrated Flora by Britton and Brown (1896-1898), which provided common names for nearly every species covered and also re- corded many local or otherwise relatively ob- scure vernacular names, was also consulted. Rafinesque’s paper (Rafinesque 1819c) on the principal features of the botany of Ken- tucky is as follows: Botany of Kentucky On its principal features, by C.S. Rafinesque, Professor of Botany and Natural History in the Transylvania University. The state of Kentucky being situated in the centre of the western country, has a flora sim- ilar to the generality of the western states and participating in their peculiar features, while it offers in itself a complete specimen of the western botany. The peculiarities of this botany consist prin- cipally in the total want of the maritime and mountains regions, which form such remark- able sections in the local floras of the Atlantic states, and abound with plants peculiar to themselves. Another striking feature in the vegetation of Kentucky and the western states is the propensity which many plants and trees exhibit of growing in a social state, to the al- most total exclusion of every other. There are many plants which grow crowded together, all over the United States; such for instance as the grasses, ferns, the Comptonia, the Stud- fonia {Hudsonia}, &c. but they allow many other plants to grow with them; while, in the western country, many extensive spaces of ground are covered with one or a few crowded species, to the exclusion of many others, which are found in their company elsewhere. The plants which may be quoted as a striking in- stance of this singular fact are not few, among which I shall select the following: (1) Vernonia prealta, Iron Weed, [Vernonia praealta Michx.; V. gigantea (Walter) Trel. ssp. gigantea; tall ironweed] (2) Baptisia cerulea, Blue Wild Indigo, [Baptisia coerulea Eaton & Wright; B. australis (L.) R.Br. ex W.T. Aiton; blue false-indigo] (3) Cacalia reniformis, Kidney Weed, [Cacalia reniformis Muhl. ex Willd.; C. muhlenber- gii (Sch. Bip.) Fern; Aroglossum muehlenbergii (Sch. Bip.) H. Rob.; great Indian-plantain; see Ap- pendix 2, entry 1, for discussion of nomenclature of this species] (4) Stedeoma pulegivides {Hedeoma_pule- gioides}, Penny-aoyal {Penny royal}, [Hedeoma pulegioides (L.) Pers.; American penny- royal, bastard pennyroyal, blue-curls] (5) Chenopodium anthelminthicum, Worm Weed, [Chenopodium anthelminticum L.; C. ambrosioides L. var. anthelminticum (L.) A. Gray; wormseed; see Appendix 2, entry 2, for discussion of nomenclature of this species] (6) Elephantopus scaber, Elephant’s Foot, [Elephantopus scaber L.; E. carolinianus Raeusch.; leafy elephant’s-foot; see Appendix 2, entry 3, for discussion of nomenclature of this species] (7) Gillenia stipulacea, Indian Physic, [Gillenia stipulata (Muhl. ex Willd.) Nutt.; Porter- anthus stipulatus (Muhl. ex Willd.) Britton; Ameri- can ipecac] (8) Miagia arupedinaria {Miegia arundinar- ia}, Cane, &c.&c. [Miegia arundinaria Raf., nom. nud., presumably intended orthography; Arundinaria gigantea (Wal- ter) Muhl.; giant cane] I consider the state of Kentucky as divided into four natural sections, or botanical regions, which are all distinguished by some peculiar- ities in their vegetation. They are: 1. THe Fuuviatite Recion. This includes all the valleys, and bottoms of the large rivers, Rafinesque Common Names—Stuckey and Pringle 13 such as the Ohio, Mississippi, Tennessee, Cumberland, Kentucky, &c. with their tribu- tary streams. The bottoms of the valleys are formed of an alluvial soil, or the washings from the hills. They are level and often overflowed: while the sides of the valleys are steep, craggy, and composed of limestone, sandstone, or sla- ty rocks. The following are some of the trees and plants peculiar to this region, and giving a decided character to its vegetation: (9) Platanus occidentalis, Sycamore or But- ton wood, [Platanus occidentalis L.; sycamore, buttonwood] (10) Hesperis pinnatifida, Ohio Wall Flower, [Hesperis pinnatifida Michx.; lodanthus pinnatifidus (Michx.) Steud.; purple rocket] (11) Jeffersonia cinata {Jeffersonia binata}, Tavin Weed {Twin leaf}, [Jeffersonia binata Barton; J. diphylla (L.) Pers.; twinleaf] (12) Capraria multifida, Sand Ragweed, [Capraria. multifida Michx., Leucospora multifida (Michx.) Nutt.; see Appendix 2, entry 4, for discus- sion of nomenclature of this species] (13) Solanum Carolinianum, Sand Briar [An orthographic variant of Solanum carolinense L., possibly preferred and originated by Rafinesque; horse-nettle] (14) Lupatorium calutinum {Eupatorium coelestinum}, Sy-weed {Sky weed}, [Eupatorium coelestinum L.; mist-flower] (15) Polanina {Polanisia} graveolens, Stink- ing weed, [Polanisia graveolens Raf.; Polanisia dodecandra (L.) DC. ssp. dodecandra; clammy-weed] (16) Heliotropium Indicum, Heliotrope, {Heliotropium indicum L.; Indian heliotrope, turn- sole; see Appendix 2, entry 5, for discussion of no- menclature of this species] (17) Catalpium cordata, Catalpa tree, [Catalpium cordifolium (J.St.-Hil.) Raf.; Catalpa speciosa (Warder ex Barney) Engelm.; northern ca- talpa; see Appendix 2, entry 6, for discussion of no- menclature of this species] (18) Populus angulata, Cotton tree, [Populus angulata Aiton; P. deltoides Marshall var. deltoides; cottonwood] (19) Porcelia tribuba {Porcelia triloba}, Pa- paw tree, [Porcelia triloba (L.) Pers.; Asimina triloba (L.) Dunal; pawpaw] (20) Synandra grandiflora, Cow mint, [Synandra grandiflora Nutt.; S. hispidula (Michx.) Baill.; see Appendix 2, entry 7, for discussion of no- menclature of this species] (21) Nelumbium pentapetalum, Swamp lily, [Nelumbium pentapetalum (Walter) Willd.; Nelum- bo lutea (Willd.) Pers.; American lotus, yellow lo- tus]; see Appendix 2, entry 8, for discussion of no- menclature of this species] (22) Pancratium liviosone {Pancratium lir- iosme}, Lily, [Pancratium liriosme Raf.; Hymenocallis caroliniana (L.) Herbert; spider-lily; see Appendix 2, entry 9, for discussion of nomenclature of this species] (23) Iris crocea, Red lily, [Iris crocea Raf., nomen nudum; presumably I. ful- va Ker Gawl.; copper iris] (24) Houstonia fruticosa, Rock weed, [Houstonia fruticosa Raf., nomen nudum; H. nigri- cans (Lam.) Fernald var. nigricans; narrow-leaved houstonia; see Appendix 2, entry 10, for discussion of nomenclature of this species] (25) Prunus pendula, Cliff plumb, &c.&c. [Prunus pendula Raf., nomen nudum, non Maxim. nec K.Koch; not definitely identified, probably P. munsoniana W. Wight & Hedrick; wild plum; see Appendix 2, entry 11, for discussion of nomencla- ture of this species] These two last are new species from the cliffs of the Kentucky river. 2. THe Centra REcion. It is formed by the limestone tract included between the val- ley of the Ohio and the hilly ridges or knobs. The ground is slightly broken, very fertile and mostly under cultivation. This section is re- markably poor in the number of botanical spe- cies growing spontaneously; I conceive that its flora hardly contains 500 species, including trees, shrubs, and naturalized plants! There are hardly any species peculiar to it; but the following ones, rare elsewhere, are here very common: (26) Eupatorium urticefolium, White nettle, [Eupatorium urticifolium Reichard, “urticaefolium”; E. rugosum Houtt.; white snakeroot] (27) Pavia muricata, Prickly Buck-eye, [Pavia muricata Raf., nomen nudum; Aesculus gla- bra Willd. var. glabra; Ohio buckeye] 14 Transactions of the Kentucky Academy of Science 58(1) (28) Isanthus ceruleus, Blue Penny-royal, [Rafinesque’s preferred orthography for Isanthus coeruleus Michx.; I. brachiatus (L.) BSP.; False pen- nyroyal | (29) Polymnia uvedalia, Scented Sun flower, [Polymnia uvedalia L.; large-flowered leafcup, yel- low-flowered leafcup] (30) Phlox glaberrima, Pink, &c.&c. [Phlox glaberrima L.; smooth phlox] It is also highly singular that in this region, the woods are open as parks, without shrubs and with very few plants, except grass or some social weeds. 3. THe Hiniy Recion. It contains the hills and ridges which divide the waters of the Ken- tucky, Green, Licking, Cumberland and Sandy rivers, &c. being spurs from the Cumberland mountains. Those hills are often called knobs, although they have not always the knobby or rounded appearance. The rocks are limestone, or sandstone, or slate. The vegetation approx- imates exceedingly to that of Virginia and Pennsylvania. On the Cumberland mountain and the highest ridges, I am told that there is a similarity with the Alleghany regions, and that the Kalmia latifolia, Common Laurel, and the Gaultheria procumbens, Mountain Tea, grow there; but having not yet visited them, I am unable to ascertain whether they ought to form another distinct region, which might be called the mountain region. The hilly region is rich in plants; I shall mention a few of those peculiar to it in Kentucky: (31) Iris cristata, Crested Tris or Flag, [Iris cristata Aiton; crested iris, crested dwarf iris] (32) Stylvianthes {Stylosanthes} elatior, Yel- low Pea-clover, [Stylosanthes elatior Raf., nomen nudum; S. biflora (L.) Britton et al.; pencil-flower] (33) Orchis ciliaris, Yellow-bunch, [Orchis ciliaris L.; Platanthera ciliaris (L.) Lindl.; orange fringed-orchid, yellow fringed-orchid] (34) Juniperus Virginiana, Red Cedar, [Juniperus virginiana L.; red cedar] (35) Vaccinium album, Wild Currant, [Vaccinium album Pursh, non L. nec Lam.; V. stam- ineum L.; deerberry] (36) Pinus rigida, Pitch Pine, [Pinus rigida Mill.; pitch pine] (37) Lechea minor, Pin week, [Lechea minor L.; small pinweed] (38) Rudbeckia fulgida, Rough Wort, [Rudbeckia fulgida Aiton, presumably var. fulgida; orange coneflower, eastern coneflower] (39) Gerardia glabrata, Yellow Wort, [Gerardia glabrata Raf., nomen nudum; not defi- nitely identified, probably Aureolaria laevigata (Raf.) Raf.; Appalachian false-foxglove; see Appendix 2, entry 12, for discussion of nomenclature of this spe- cies| (40) Asarum Virginicum, Heart-leaf, &c.&c. [Asarum virginicum L.; Hexastylis virginica (L.) Small; heart-leaf, little brown jugs; see Appendix 2, entry 13, for discussion of identity of this species] 4. THe BarrREN REGION, or rather the open region. This has an extensive range in Kentucky, particularly in the western and southern parts of the state. The numerous barrens and licks compose it, lying scattered and irregularly among the central and hilly regions. The barrens are tracts of ground des- titute of trees, or with few scattered small ones; but thickly covered with a luxuriant growth of plants; while the licks are almost destitute of them, and those that grow in their immediate neighbourhood are all small, which is owing to their poor, slaty or argilla- ceous soil. Their vegetation is however simi- lar to that of the barrens. Both have a growth of plants very similar to the vegetation of the prairies of Ohio, Indiana, and Illinois, and more different from that of the Atlantic states, than the three foregoing regions. The plants peculiar to them are very numerous; I shall mention only a few, among the most re- markable and singular. (41) Solidago rigida, Stiff Golden-rod, [Solidago rigida L.; stiff goldenrod] (42) Polygala polygama, Nimble weed, [Polygala polygama Walter; racemed milkwort, bit- ter milkwort] (43) Rudbeckia purpurea, Purple Sun-flow- eX, [Rudbeckia purpurea L.; Echinacea purpurea (L.) Moench; purple coneflower] Rafinesque Common Names—Stuckey and Pringle 15 (44) Ruellia oblongifolia, Rough Bell, [Ruellia oblongifolia Raf., nomen nudum, Raf. ex Nees, pro syn., non Michx.; R. caroliniensis (J.F. Gmel.) Steud. ssp. ciliosa (Pursh) R.W. Long; hairy ruellia] (45) Andropogon arenaceum, Barren Oats, [Andropogon avenaceus Michx.; Sorghastrum nu- tans (L.) Nash; Indian grass] (46) [Andropogon] nutans, Barren Oats, [Andropogon nutans L.; Sorghastrum nutans (L.) Nash; Indian grass] (47) Petalvitemon {Petalostemon} candidum, Nimble clover, [Petalostemon candidum (Michx. ex Willd.) Michx.; Dalea candida Michx. ex Willd.; white prairie-clo- ver| (48) [{Petalosteum}] purpureum, Nimble clover, [Petalostemon purpureum (Vent.) Rydb.; Dalea pur- purea Vent. var. purpurea; purple prairie-clover] (49) Silphium therebinthaceum, Turpentine weed, [Presumably an intentional abridgement of Sil- phium terebinthinaceum Jacq.; prairie-dock] (50) Silene catesbri {Silene catesbei}, Scarlet Pink, [Silene catesbaei Walter; S. virginica L.; fire-pink] (51) Gentiana amarellvides {Gentiana amar- elloides}, Yellow Gentian, [Gentiana amarelloides Michx.; Gentianella quin- quefolia (L.) Small var. quinquefolia; stiff gentian; see Appendix 2, entry 14, for discussion of nomen- clature of this species] (52) Buchnera Americana, Black Wort, &c.&e. [Buchnera americana L.; blue-hearts] From the above a faint, but correct idea may be formed of the display and peculiarities of the wide range of vegetation in Kentucky and throughout the western states, wherein the same peculiar divisions or regions may be traced. The vulgar names of the plants above men- tioned are such as I found used in some parts of Kentucky; but they cannot claim to be gen- erally understood even in this state, many be- ing merely local or personal. The botanical names are alone to be relied on, being uni- versal and not liable to mislead. APPENDIX 1: Indexes to Botanical and Common Names of Plants as Originally Listed and Those Added to Rafinesque’s 1819 Paper on the Botany of Kentucky. A. Botanical or Scientific Names Aesculus glabra Willd. var. glabra (27) Andropogon arenaceum (45) Andropogon avenaceus Michx. (45) Andropogon nutans L. (46) Aroglossum muehlenbergii (Sch. Bip.) H. Rob. (3) Arundinaria gigantea (Walter) Muhl. (8) Aureolaria laevigata (Raf.) Raf. (39) Asarum virginicum L. (40) Asimina triloba (L.) Dunal (19) Baptisia australis (L.) R. Br. ex W.T. Aiton (2) Baptisia cerulea (2) Baptisia coerulea Eaton & Wright (2) Buchnera americana L. (52) Cacalia muhlenbergii (Sch. Bip.) Fernald (3) Cacalia reniformis Muhl. ex Willd. (3) Capraria multifida Michx. (12) Catalpa speciosa (Warder ex Barney) Engelm. (17) Catalpium cordata (17) Catalpium cordifolium (J. St.-Hil.) Raf. (17) Chenopodium anthelminthicum L. (5) Chenopodium ambrosioides L. var. anthelminticum (L.) A. Gray (5) Dalea candida Michx. ex Willd. (47) Dalea purpurea Vent. var. purpurea (48) Echinacea purpurea (L.) Moench (43) Elephantopus carolinianus Raeusch. (6) Elephantopus scaber L. (6) Eupatorium celestinum (14) Eupatorium coelestinum L. (14) Eupatorium rugosum Houtt. (25) Eupatorium urticefolium (26) Eupatorium urticifolium Reichard (26) Gentiana amarelloides Michx. (51) Gentiana amarellvides (51) Gentiana quinquefolia (L.) Small var. quinquefolia (51) Gerardia glabrata Raf. (39) Gillenia stipulacea (7) Gillenia stipulata (Muhl. ex Willd.) Nutt. (7) Hedeoma pulegioides (L.) Pers. (4) Heliotropium indicum L. (16) Hesperis pinnatifida Michx. (10) Hexastylis virginica (L.) Small (40) Houstonia fruticosa Raf. (24) Houstonia nigricans (Lam.) Femald var. nigricans (24) Hymenocallis caroliniana (L.) Herbert (22) Iodanthus pinnatifidus (Michx.) Steud. (10) Iris cristata Aiton (31) Iris crocea Raf. (23) 16 Transactions of the Kentucky Academy of Science 58(1) Iris fulva Ker Gawl. (23) Stylvianthes elatior (32) Isanthus brachiatus (L.) BSP. (28) Synandra grandiflora Nutt. (20) Isanthus caeruleus Michx. (28) Synandra hispidula (Michx.) Baill. (20) Isanthus ceruleus (28) Vaccinium album Pursh (35) Jeffersonia binata (11) Vaccinium stamineum L. (35) Jeffersonia cinata (11) Vernonia gigantea (Walter) Trel. Jeffersonia diphylla (L.) Pers. (11) ssp. gigantea (1) Juniperus virginiana L. (34) Vernonia praealta Michx. (1) Lechea minor L. (37) Vernonia prealta (1) Leucospora multifida (Michx.) Nutt. (12) Lupatorium calutinum (14) Miagia arupedinaria (8) Miegia arundinaria Raf. (8) Nelumbium pentapetalum (Walter) Willd. (21) Ammericaublotas OD) Nelumbo lutea (Willd.) Pers. (21) American pennyroyal (4) Orchis ciliaris L. (33) Barren oats (45, 46) Pancratium liriosme Raf. (22) B. Index to Common or Vulgar Names Appalachian false-foxglove (39) American ipecac (7) Bastard pennyroyal (4) Pancratium liviosone (22) Bitter milkwort (42) Pavia muricata Raf. (27) Black wort (52) Petalostemon candidum (Michx. ex Willd.) Michx. (47) Blue-eurs(4) Petalostemon purpureum (Vent.) Rydb. (48) Blue-hearts (52) Petalvitemon candidum (47) Blue penny-royal (28) Phlox glaberrima L. (30) Blue false-indigo (2) Pinus rigida Mill. (36) Blue wild indigo (2) Platanus occidentalis L. (9) Button wood (9) Platanthera ciliaris (L.) Lindl. (33) Buttonwood (9) Polanina graveolens (15) Cane (8) Polanisia graveolens Raf. (15) Catalpa tree (17) Polanisia dodecandra (L.) DC. Clammy-weed (15) ssp. dodecandra (15) Cliff plumb (25) Polygala polygama Walter (42) Copper iris (23) Polymnia uvedalia L. (29) Cotton tree (18) Populus angulata Aiton (18) Cottonwood (18) Populus deltoides Marshall Cow mint (20) var. deltoides (18) Crested dwarf iris (31) Porcelia tribuba (19) Crested flag (31) Porcelia triloba (L.) Pers. (19) Crested iris (31) Porteranthus stipulatus (Muhl. ex Willd.) Britton (7) Crested tris (31) Prunus munsoniana W. Wight & Hedrick (25) Deerberry (35) Prunus pendula Raf. (25) Eastern coneflower (38) Ruellia caroliniensis (J.F. Gmel.) Steud. Elephant’s foot (6) ssp. ciliosa (Pursh) R.W. Long (44) False pennyroyal (28) Ruellia oblongifolia Raf. (44) Fire-pink (50) Rudbeckia fulgida Aiton (38) Flag (31) Rudbeckia purpurea L. (43) Giant cane (8) Silene catesbaei Walter (50) Great Indian-plantain (3) Silene catesbei (50) Hairy ruellia (44) Silene catesbri (50) Heart-leaf (40) Silene virginica L. (50) Heliotrope (16) Silphium terebinthaceum (49) Horse nettle (13) Silphium terebinthinaceum Jacq. (49) Indian grass (45, 46) Solanum carolinianum (13) Indian heliotrope (16) Solanum caroliniense L. (13) Indian physic (7) Solidago rigida L. (41) Iron weed (1) Sorghastrum nutans (L.) Nash (45, 46) Kidney weed (3) Stedeoma pulegioides (4) Large-flowered leafcup (29) Stylosanthes biflora (L.) BSP. (32) Leafy elephant’s-foot (6) Stylosanthes elatior Raf. (32) Lily (22) Rafinesque Common Names—Stuckey and Pringle 17 Little brown jugs (40) Mist-flower (14) Narrow-leaved houstonia (24) Nimble clover (47, 48) Nimble weed (42) Northern catalpa (17) Ohio buckeye (27) Ohio wall flower (10) Orange coneflower (38) Orange fringed-orchid (33) Pawpaw (19) Pawpaw tree (19) Pencil-flower (32) Penny-aoyal (4) Penny-royal (4) Pink (30) Pin weed (37) Pitch pine (36) Prairie-dock (49) Prickly buck-eye (27) Purple coneflower (43) Purple prairie-clover (48) Purple rocket (10) Purple sun-flower (43) Racemed milkwort (42) Red cedar (34) Red lily (23) Rock weed (24) Rough bell (44) Rough wort (38) Sand briar (13) Sand ragweed (12) Scarlet pink (50) Scented sun flower (29) Sky-weed (14) Small pinweed (37) Smooth phlox (30) Spider-lily (22) Stiff gentian (51) Stiff golden-rod (41) Stiff goldenrod (41) Stinking weed (15) Swamp lily (21) Sycamore (9) Sy-weed (14) Tall ironweed (1) Tavin weed (11) Turnsole (16) Turpentine weed (49) Twinleaf (11) Twin weed (11) White nettle (26) White prairie-clover (47) White snakeroot (26) Wild currant (35) Wild plum (25) Wormseed (5) Worm weed (5) Yellow-bunch (33) Yellow-flowered leafcup (29) Yellow fringed-orchid (33) Yellow gentian (51) Yellow lotus (21) Yellow pea-clover (32) Yellow wort (39) APPENDIX 2: Notes on the Botanical and Common Names (by J.S.P.) 1. The nomenclature of this species is unsettled at the time of this writing, not only because of differences of opinion as to appropriate generic circumscriptions, but also pending a decision on proposals to reject the name Cacalia, or, alternatively, for conserved typification. If Ca- calia is rejected or is typified otherwise than as noted be- low, this species will presumably be placed in Arnoglossum regardless of whether the genus is circumscribed so as to include Synosma Raf. ex Britton & A. Brown; if Cacalia is not rejected, but is conserved with C. atriplicifolia L. as the type, the correct name for this species will be Ca- calia muehlenbergii (Sch. Bip.) Fernald. 2. The use of “wormseed” in works by other authors from the same period indicates that “worm weed” prob- ably represents a typographical error rather than a version used in Kentucky during Rafinesque’s time. 3. The name Elephantopus scaber L. remains the cor- rect name for an accepted species, but not for a species that occurs in Kentucky. Elephantopus scaber sensu Mi- chaux has been identified as the species now called E. carolinianus. 4. No common name has been located for Leucospora multifida in recent references other than the generic name, or the now-obsolete generic name Conobea, used as such. If the use of “sand-ragweed” actually persists, it might be revived for wider application, hyphenated be- cause this species is not in the ragweed genus Ambrosia (Asteraceae). If, however, this was a coinage that expired with Rafinesque, a name that might seem to indicate a relationship to the true ragweeds would not be ideal. 5. Heliotropium indicum L. is now extensively but spo- radically adventive or naturalized in the southeastern United States, and, despite Kentucky's inland location, it is not inconceivable that Rafinesque might have found a small population there as early as 1819. However, because no other North American authors mentioned H. indicum as a naturalized species until considerably later, and be- cause to the present day H. indicum has remained spo- radic in its North American occurrences, it hardly seems credible that Rafinesque could have found this species in such abundance that it was among those “giving a decided character to [the] vegetation” of any part of Kentucky. Rafinesque’s (1838) later description of the species he had so identified, which he then called Elopia riparia Raf., appears to have been derived largely from published de- scriptions of H. indicum, but inevitably one wonders if his concept of the species was actually based on some other boraginaceous species. 18 Transactions of the Kentucky Academy of Science 58(1) 6. Rafinesque presumably referred to the Catalpa spe- cies that is native to Kentucky, viz. C. speciosa (Warder ex Barney) Engelm., which he might have seen on his trip to the mouth of the Ohio River in 1818. There seems to be an error in Index Rafinesquianus (Merrill 1949) on this point. It is probably appropriate to assume from the sim- ilarity of the epithets, as Merrill evidently did, that Ca- talpium cordatum Raf. (Rafinesque 1819c) was intended to be the same as his C. cordifolium Raf. (Rafinesque 1819b), published slightly earlier, the second being either a lapsus calami or an intentionally amended orthography for the first. According to Merrill, Catalpium cordifolium Raf. was tied nomenclaturally to Catalpa cordifolia J. St.- Hil. Merrill apparently confused C. cordifolia J. St.-Hil. of 1804 with C. cordifolia Moench of 1794, which is a synonym of C. bignonioides (Rehder 1949). The illegiti- mate homonym Catalpa cordifolia J. St.-Hil., however, is a synonym of C. speciosa, according to Rehder; this fits well with Rafinesque’s report of his Catalpium cordifolium/C. cordatum from Kentucky. 7. We have located no common name for Synandra hispidula in recent literature, except for the generic name used as such. As “cow-mint” does not appear to be used for any other species, it could be considered for this spe- cies, which is in the mint family (Lamiaceae). 8. Some recent authors have advocated “lotus-lily” or “water-lotus” for Nelumbo, presumably because Lotus is the botanical name for a genus in the Fabaceae. With this genus very generally being known simply as “lotus,” this illustrates the recurrent question as to what extent stan- dardized English-language names should vary from names actually in common usage. In this case, one might also ask whether “lotus-lily,” even if hyphenated, is all that much preferable for standardization, since Nelumbo is not in the Liliaceae. 9. The name Pancratium liriosme Raf., published 2 years earlier by Rafinesque (1817c), is the basionym of Hymenocallis liriosme (Raf.) Shinners. The species thus named is native to Arkansas, Louisiana, Oklahoma, and Texas. According to Shinners (1951), Rafinesque (1817c), in describing P. liriosme, “was relying on [Claude C.] Ro- bin’s description of plants that the latter had observed wild in Louisiana.” The only species of Hymenocallis native to Kentucky is H. caroliniana (L.) Herb., formerly known as H. occidentalis (Leconte) Kunth. 10. No satisfactory common name appears to be avail- able for this species. In popular field guides, the generic name Houstonia is used as a common name for species of this genus, except for two species of different aspect that are called “bluets.” Britton and Brown (1896-1898) called this species “narrow-leaved houstonia,” when it was known botanically H. angustifolia Michx., but in the pop- ular field guide by Newcomb (1977), “narrow-leaved houstonia” is used for H. tenuifolia Nutt. Complicating the issue, some botanists prefer to include Houstonia in Hed- yotis. The name madderwort has reportedly been applied to Houstonia, but this may have been in a context extend- ing to other genera in the Rubiaceae (madder family). “Rock weed” has little to recommend it, having been ap- plied to Asperula odorata L. and possibly other species. 11. From the habitat given by Rafinesque—‘cliffs of the Kentucky River’—an anonymous reviewer has con- cluded that Rafinesque’s Prunus pendula is probably P. munsoniana W. Wight & Hedrick, a species that was not otherwise recognized as distinct and given a scientific name until 1911. Under the current rules of botanical no- menclature, because Rafinesque provided no description of his “new species,” and because another botanist had used the name Prunus pendula for a different species 4 years earlier, this interpretation does not affect the no- menclature of P- munsoniana. 12. On the basis of Rafinesque’s phytogeographic com- ments, Aureolaria laevigata, “Appalachian false-foxglove,” seems the most likely identity of this species, but A. flava (L.) Farw., “smooth false-foxglove,” cannot be ruled out. 13. Rafinesque’s “outline of four vegetation regions of Kentucky” was published before its author visited that portion of Kentucky in which Hexastylis species occur; for this reason it has been suggested that his Asarum virgin- icum might not have been any species now referred to Hexastylis, the alternative interpretation being that it was actually A. canadense L. This seems unlikely because Raf- inesque presumably would have been familiar with A. can- adense under its correct name and because, in all of the literature available prior to 1819 in which A. virginicum is described, that species is said to have glabrous, mottled leaves. (Similarly, Rafinesque had not at that time traveled into areas of Kentucky where any species of Lechea is native, but one would hardly assume that he had misiden- tified some other genus as Lechea.) Even if Rafinesque’s concept of the “Hilly Region” included the Knobs that are east of Lexington but west of the Pottsville escarpment, it seems likely that some of his information, at least in this relatively brief section of the paper, was more or less sec- ondhand, perhaps derived from a combination of accounts from other naturalists, studies of specimens in their her- baria, and extrapolations from his firsthand knowledge of the flora of adjacent states. 14. The name Gentiana amarelloides Michx. is a het- erotypic synonym of Gentianella quinquefolia (L.) Small var. quinquefolia, but in Kentucky this species is repre- sented by var. occidentalis. From the Medical Flora, it is evident that Rafinesque (1828) considered the plants with yellow corollas to be a different species from the more widespread form with purple corollas. LITERATURE CITED Britton, N.L., and A. Brown. 1896-1898. An illustrated flora of the northern United States, Canada and the British possessions .... Charles Scribner's Sons, New York, NY. Femald, M.L. 1950. Gray's manual of botany. 8th ed. American Book Company, New York, NY. Reprinted 1970, D. Van Nostrand Co., New York, NY. Gleason, H.A., and A. Cronquist. 1991. Manual of vas- cular plants of northeastern United States and adjacent Rafinesque Common Names—Stuckey and Pringle 19 Canada. 2nd ed. New York Botanical Garden, New York, NY. McAtee, W.L. 1913-1933. Some local names of plants, I_-V. Torreya 13:225-236 (1913); 16:235—-242 (1916); 20: 17-27 (1920); 26:3-10 (1926); 33:81-86 (1933). Merrill, E.D. 1949. Index Rafinesquianus: the plant names published by C.S. Rafinesque with reductions, and a consideration of his methods, objectives, and at- tainments. Arnold Arboretum of Harvard University, Ja- maica Plain, MA. Newcomb, L. 1977. Newcomb’s wildflower guide. Little, Brown and Company, Boston, MA. Peterson, R.T., and M. McKenny. 1968. A field guide to wildflowers of northeastern and north-central North America. Houghton Mifflin Company, Boston, MA. Rafinesque, C.S. 1811. Botanical information concerning two families of plants. I. Species of the genus Callitri- che. II. North American species of the genus Potamo- geton. Med. Repos., third hexade 2(4):407—409. Rafinesque, C.S. 1817a. [Review of Amos Eaton’s] A manual of botany for the northern states, comprising generic descriptions of all phenogamous and cryptoga- mous plants to the north of Virginia. Am. Monthly Mag. & Crit. Rev. 1(6):426-430. Rafinesque, C.S. 1817b. Florula of the White Mountain of New-Hampshire. Am. Monthly Mag. & Crit. Rev. 1(6):440-442. Rafinesque, C.S. 1817c. Florula Ludoviciana: or, a flora of the state of Louisiana. C. Wiley & Co., New York, NY. Facsimile ed. 1967, with introduction by Joseph Ewan, Hafner Publishing Co., New York, NY. Rafinesque, C.S. 1819a. Prodrome des nouveaux genres de plantes observés en 1817 et 1818 dans l’intérieur des états-Unis d’Amérique. J. Phys. Chim. Hist. Nat. Arts 89(2):96-107. Rafinesque, C.S. 1819b. [Review of] The genera of North-American plants and a catalogue of the species to the year 1817. By Thomas Nuttall, F.L.S. &c. 2 vols. 12 mo. Philadelphia, PA. 1818. Am. Monthly Mag. & Crit. Rev. 1(3):184—196. Rafinesque, C.S. 1819c. Botany of Kentucky[:] on its principal features. Western Rev. & Misc. Mag. 1(2):92- 95. Rafinesque, C.S. 1819d. Errata [to botany of Kentucky]. Western Rev. & Misc. Mag. 1(3):128. Rafinesque, C.S. 1820. Prodrome d’une monographie des rosiers del Amérique septentrionale, contenant la de- scription de quinze nouvelles espéces et vingt variétés. Ann. Gén. Sci. Phys. 5(14):210-220. Rafinesque, C.S. 1824. Florula Kentuckensis. Catalogue of the principal trees, shrubs and plants of Kentucky. Pages [12]-16 in First catalogues and circulars of the Botanical Garden of Transylvania University at Lexing- ton in Kentucky, for the year 1824. Printed for the Bo- tanical Garden Company by John M. M’Calla, Lexing- ton, KY. Rafinesque, C.S. 1828. Medical flora; or, Manual of the medical botany of the United States of North America ., volume the first. Atkinson & Alexander, Philadel- phia, PA. Rafinesque, C.S. 1830. Medical flora; or Manual of the medical botany of the United States of North America ..., volume the second. Samuel C. Atkinson, Philadel- phia, PA. Rafinesque, C.S. 1836-[1838]. New flora of North Amer- ica. Published by the author, Philadelphia, PA. Facsim- ile ed. 1946, Arnold Arboretum, Jamaica Plain, MA. Rafinesque, C.S. “1836” [1837-1838]. Flora Telluriana. Published by the author, Philadelphia. Facsimile ed. 1946, Arnold Arboretum, Jamaica Plain, MA. Rafinesque, C.S. 1838. Sylva Telluriana. Published by the author, Philadelphia. 184 pp. Facsimile ed. 1943, Ar- nold Arboretum, Jamaica Plain, MA. Rafinesque, C.S. 1840. Autikon botanikon or botanical illustrations .... Published by the author, Philadelphia, PA. Facsimile ed. 1942, Armold Arboretum, Jamaica Plain, MA. Rehder, A. 1949. Bibliography of cultivated trees and shrubs hardy in the cooler temperate regions of the northern hemisphere. Arnold Arboretum of Harvard University, Jamaica Plain, MA. Shinners, L.H. 1951. The north Texas species of Hymen- ocallis (Amaryllidaceae). Field & Lab. 19:102-104. Trans. Ky. Acad. Sci. 58(1):20—22. 1997. Some Comments on Constantine Rafinesque’s 1819 Description of Botanical Regions in Kentucky William S. Bryant Department of Biology, Thomas More College, Crestview Hills, KY 41017 Perhaps of greatest ecological significance in Rafinesque’s (1819a, 1819b) Botany of Ken- tucky was his recognition of distinct botanical regions or natural sections in the state. His regions and plant associations were not so well defined as those currently recognized (e.g., Braun 1950: Kiichler 1964; Quarterman and Powell 1978), but at least they were a first at- tempt. Meijer (1973) noted that Rafinesque often had good ideas of natural relationships of vegetation as distinguished by its peculiar- ities. Rafinesque (1819a) recognized four botan- ical regions but suggested that a fifth, the Mountain Region, was probable. He had nei- ther collected in nor visited the eastern por- tions of Kentucky. His entrance into Kentucky was from Pennsylvania via the Ohio River in 1818. From 1819-1825, he was Kentucky's first professional resident botanist, teaching at Transylvania College in Lexington (Meijer 1973). It seems apparent that Rafinesque was familiar with the literature concerning some aspects of natural history on the frontier as detailed by Drake (1815), Filson (1784), Imlay (1797), and Michaux (1904). Kentucky was only 26 years old as a state when Rafinesque arrived in 1818 and only 44 years removed from the establishment of its first permanent settlement. Rafinesque’s botanical regions of Kentucky are listed and described as follows: 1. The Fluviatile Region included the val- leys and bottoms of large rivers and their trib- utaries. Rafinesque especially noted the allu- vial deposits but also included the steep-sided gorges bordering many of these streams. His inclusion of the Cumberland and Kentucky rivers might be expected, but his inclusion of the Tennessee River, which prior to 1818 formed the state’s western border, and the Mississippi River, which after the 1818 Jack- son Purchase became the western border, sug- gests that Rafinesque was abreast of happen- ings in the Commonwealth. His recognition of 20 a Fluviatile Region does not fit most modern regional designations; however, Middleton et al. (1926) stated that “the Alluvial land of the Mississippi River bottoms constitutes the fifth distinct area of the state.” The Ohio River low- lands was also designated as a distinct geo- graphical region (Burroughs 1926). Cotterill (1917) noted that early maps of Kentucky “seemed a mere network of rivers” (Figure 1). Rafinesque’s list of plants and trees peculiar to this region was limited. He included syca- more (Platanus occidentalis) and cottonwood (Populus deltoides), which are widely distrib- uted throughout the state, but his inclusion of catalpa (Catalpa speciosa) and spiderlily (Hy- menocallis caroliniana) suggests that he had some knowledge of the western third of the state where these two species are found in lowland forests. 2. The Central Region is the Bluegrass Re- gion, which is bordered by the Knobs. He not- ed that this fertile limestone area was mostly under cultivation even in 1819. He also rec- ognized a low species diversity including na- tive and naturalized plants. Meijer (1973) pointed to Rafinesque’s interest in the flora of introduced weeds and how this invasion co- incided with the arrival of white settlers. Rafinesque’s statement that “it is also highly singular that in this region, the woods are open as parks, without shrubs and with very few plants, except grass or some social weeds,” is significant. He was describing the blue ash- oak savanna woodland, which was the char- acteristic vegetation of the Bluegrass region (Bryant 1983; Bryant et al. 1980). His list of species in no way characterizes this region. 3. The Hilly Region included the Knobs as well as extensions of broken ground from the Big Sandy River in the northeast to the Green River in the west. The geologic substrate men- tioned by Rafinesque included sandstone, limestone, or slate. He found that the vege- tation approximated that of Pennsylvania and Virginia, with which he was familiar. The Comments on Rafinesque—Bryant Dil u to = SS Se a = SSS eee to — 7 ~ ‘ CORSET), ¢() Ye Wenpeky Bu SS ULRs. i hy COW ay tes, Figure 1. The Samuel Lewis 1814 map of Kentucky showing the principal features. Note the lack of information on the mountains of eastern Kentucky but the detail of the many streams and rivers. After Clark (1979). plants listed, especially redcedar (Juniperus virginiana), pitch pine (Pinus rigida), and deerberry (Vaccinium stamineum) are found on various substrates in the Knobs Region. Muller and McComb (1983) described the up- land vegetation of the Knobs as being domi- nated by oaks, none of which Rafinesque men- tioned. 4. The Mountain Region was included with the Hilly region, but Rafinesque acknowl- edged that he had not visited this part of the state. He had been told of the similarity of the mountains to the Allegheny region. This re- gion included the Cumberland Mountains and the highest ridges. The presence of mountain laurel (Kalmia latifolia) and mountain tea (Gaultheria procumbens) and a few of the plants listed for the Hilly Region are inade- quate to describe this area. Braun (1950) char- acterized this portion of eastern Kentucky as the center of the mixed mesophytic forest, the richest forest type in eastern North America. 5. The Barren Region encompassed the open country in the western and southern parts of the state. Rafinesque noted that the vegetation consisted of “a growth of plants very similar to that of the prairies of Ohio, Indiana, and Illinois.” He pointed out the sparsity of trees and the presence of licks, which tended to attract grazing animals (e.g., buffalo). Rafinesque’s list of plants of this re- gion is perhaps more representative than his lists for the other regions he described. Blue- stems (Andropogon spp.), Indian grass (Sor- ghastrum nutans), prairie-clovers (Petaloste- mon candidum and P. purpureum), prairie- dock (Silphium terebinthinaceum), and purple coneflower (Echinacea purpurea) are still found in barren remnants (Baskin and Baskin 1978; Braun 1950). Rafinesque was most in- terested in the Barrens; along with his collab- orator, Dr. Charles Short, he collected there extensively (Meijer 1973). Rafinesque’s Barren Region is part of the Pennyroyal or Mississippi Plateau. As can be determined from the above de- scriptions of Botanical Regions, Rafinesque in 1819 was more familiar with those portions of Kentucky from Lexington westward. Since he travelled to and kept an association with some of the residents of Harmonie (later New Har- mony) in southwestern Indiana (Thompson 1898), it seems probable that he botanized on his trips. In conclusion, it was indeed fortunate that Rafinesque made reference to different botan- ical regions as being present in Kentucky. He noted some of the plants, although not always 22. Transactions of the Kentucky Academy of Science 58(1) those that might best characterize a commu- nity or region. He mentioned topography, geo- logic substrate, and soils as being of impor- tance in regional differentiations. It is unfor- tunate that he did not continue his interest and study of plant associations at a time prior to their manipulation and fragmentation by the onrush of settlement. LITERATURE CITED Baskin, J.M., and C.C. Baskin. 1978. Plant ecology of cedar glades in the Big Barrens region of Kentucky. Rhodora 80:545-557. Braun, E.L. 1950. Deciduous forests of eastern North America. Blakiston Co., Philadelphia, PA. Bryant, W.S. 1983. Savanna-woodland in the Outer Blue- grass of Kentucky. Trans. Ky. Acad. Sci. 44:44—49. Bryant, W.S., M.E. Wharton, W.H. Martin, and J.E. Var- ner. 1980. The blue ash-oak savanna-woodland, a rem- nant of pre-settlement vegetation in the Inner Bluegrass of Kentucky. Castanea 45:149-165. Burroughs, W.C. 1926. Geography of the Knobs. Ken- tucky Geological Survey, Frankfort, KY. Clark, T.D. 1979. Historic maps of Kentucky. Univ. Press of Kentucky, Lexington, KY. Cotterill, R. 1917. History of pioneer Kentucky. Johnson & Hardin, Cincinnati, OH. Drake, D. 1815. Natural and statistical view, or picture of Cincinnati and the Miami Country. Looker and Wal- lace, Cincinnati, OH. Filson, J. 1784. The discovery, settlement, and present state of Kentucky. James Adams, Wilmington, DE. Imlay, G. 1797. A topographical description of the west- em territory of North America. Debrett, London. Kiichler, A. W. 1964. Potential natural vegetation of con- terminous United States. Am. Geogr. Soc. Spec. Publ. 64. Meijer, W. 1973. The contributions by Rafinesque to the early botanical exploration of Kentucky. Castanea 38: 261-265. Michaux, F.A. 1904. Travels to the west of the Allegheny Mountains in the states of Ohio, Kentucky and Ten- nessee, and back to Charleston, by the Upper Carolinas. Pages 110-306 in R.G. Thwaites. 1904. Early Western Travels 1746-1846. Arthur H. Clark Co., Cleveland, OH. Middleton, A.R., W.R. Jillson, FT. McFarland, and W.A. Anderson Jr. 1926. Kentucky. Pages 149-154 in V.E. Shelford (ed). Naturalist’s guide to the Americas. Wil- liams & Wilkins Co., Baltimore, MD. Muller, R.N., and W.C. McComb. 1983. Upland forests of the Knobs Region of Kentucky. Bull. Torrey Bot. Club 113:268-280. Quarterman, E., and R.L. Powell. 1978. Potential ecol- ogical/geological natural landmarks on the Interior Low Plateaus. U.S. Dept. of Interior, National Park Service. Rafinesque, C.S. 1819a. Botany of Kentucky. On its prin- cipal features. Western Rev. & Misc. Mag. 1(2):92-95. Rafinesque, C.S. 1819b. Errata [to botany of Kentucky]. Western Rev. & Misc. Mag. 1(3):128 Thompson, M. 1898. Stories of Indiana. American Book Co., New York, NY. Trans. Ky. Acad. Sci. 58(1):23-28. 1997. Effect of Light on Daily Emergence of Cercariae of the Trematodes Echinostoma trivolvis (Echinostomatidae) and Cephalogonimus vesicaudus (Cephalogonimidae) from Natural Infections of the Snail Helisoma trivolvis (Planorbidae) at Owsley Fork Reservoir, Kentucky Ron Rosen, Peter Blair, Jeff Ellington, and Jason Backu Department of Biology, Berea College, Berea, KY 40404 ABSTRACT Naturally infected snails, Helisoma trivolvis, releasing cercariae of Echinostoma trivolvis and Cephalogon- imus vesicaudus, were collected from Owsley Fork Reservoir in northern Jackson and southem Madison counties, Kentucky, in June or July 1990, 1992, and 1995. Emergence of cercariae from snails was assessed at 26° C for 24 h under the following photoperiod regimens: (1) 12 h light:12 h dark; (2) 12 h dark:12 h light (inverted photoperiod); (3) 24 h light; and (4) 24 h dark. In the split photoperiods, maximum cercarial emergence of E. trivolvis and C. vesicaudus was significantly greater in the 12 h of light and 12 h of dark, respectively. Photoperiod inversion resulted in a complete reversal in timing of cercarial release for both species. Continuous light or dark for 24 h induced continuous release of cercariae of both species. No significant change was observed in the timing of peak cercarial emergence of C. vesicaudus in the 1990, 1992, and 1995 studies using a 12 h light:12 h dark photoperiod, but a marked difference was noted for E. trivolvis in 1995 when peak cercarial release was shifted to the dark phase of the 24 h photoperiod. These results are discussed with regard to (1) exogenous vs. endogenous factors affecting cercarial release and (2) the use of cercarial emergence patterns for identifying parasitic strains. INTRODUCTION The timing of the emergence of trematode (subclass Digenea) cercariae from their snail host has been linked to activity of the next host in the life cycle in several studies (Lewis, Welsford, and Uglem 1989; Theron 1984), but the physiological triggers associated with cer- carial release are poorly understood. It has been suggested that in-vitro cultivation of in- tramolluscan stages in the absence of snail tis- sue will be necessary to clarify the exact me- chanisms/triggers (Glaudel and Etges 1973). In addition to a possible endogenous circadian rhythm, these triggers may be influenced by a number of environmental factors including temperature, humidity, oxygen, pH, and light (Smyth and Halton 1983). These exogenous factors either act directly on the parasite or are mediated indirectly through the snail host. That light triggers or inhibits cercarial re- lease from the snail host in many species of digenetic trematodes is well known. Reversal of the 24 h light:dark photoperiod (i.e., pho- toperiod inversion) often results in complete reversal in the pattern of cercarial release (Asch 1972; Giovannola 1936; Glaudel and Et- ges 1973; Luttermoser 1955; Oliver 1951; Wagenbach and Alldredge 1974). Such a re- 23 versal provides evidence that the pattern of release is not a true circadian rhythm but is mediated by the “rhythm of the snail which is controlled to some extent by illumination” (Asch 1972). However, a number of these studies did not assess whether cercarial emer- gence ceases to be rhythmic if the light or dark is of a constant magnitude (i.e., contin- uous light or dark). This criterion must be met to rule out the presence of an endogenous cir- cadian rhythm within the parasite itself (Wag- enbach and Alldredge 1974). Distinct strains of digenetic trematodes have been documented in several studies based on photoperiod and cercarial emer- gence patterns (Gumble et al. 1957; Riley and Uglem 1995; Theron 1984), but, to our knowl- edge, shifting patterns of cercarial emergence in a species in the same habitat over time have not been observed. Further studies of this na- ture might provide (1) insight into use of cer- carial emergence patterns as stable biological characteristics for some species of digenetic trematodes and (2) a method for identifying different strains of a species from the same habitat. Little information is available concerning the effect of light on emergence of cercariae of Echinostoma trivolvis (Echinostomatidae) 24 Transactions of the Kentucky Academy of Science 58(1) (Schmidt and Fried 1996), and no studies of this nature have been conducted with cercari- ae of Cephalogonimus vesicaudus (Cephalo- gonimidae). Both species are parasites of the snail Helisoma trivolvis (Planorbidae) at Ows- ley Fork Reservoir in northern Jackson and southern Madison counties, Kentucky (Rosen et al. 1994). Thus, the objectives of our study were to determine (1) the effect of light on the release of E. trivolvis and C. vesicaudus cercariae from natural infections of H. trivol- vis and (2) whether different strains of these species exist in the reservoir based on photo- period and cercarial emergence patterns as- sessed over several years. MATERIALS AND METHODS Specimens of H. trivolvis were collected from Owsley Fork Reservoir in June or July 1990, 1992, and 1995. A brief description of the study site is provided in Rosen et al. (1994). Within 2 h of their collection, snails were placed individually into 50 ml beakers filled with 40 ml of filtered reservoir water, incubated at 26° C with a 12 h light (0700- 1859):12 h dark (1900-0659) cycle, and screened for cercarial emergence at 1400, 2030, and 0830 for 1 day. Snails with mature infections (ie., releasing cercariae of E. tri- volvis or C. vesicaudus) were then immediate- ly acclimated for 24 h at 26° C to one of the following photoperiod regimens: (1) 12 h light (0700-1859):12 h dark (1900-0659), (2) 12 h dark (0700-1859):12 h light (1900-0659), (3) 24 h light, and (4) 24 h dark. Only the 12 h light:12 h dark cycle was used to compare emergence patterns in the 1990, 1992, and 1995 studies. Following the initial 24 h acclimation peri- od, cercarial counts were made every 2 or 4 hours for 24 h. At each time interval, snails were transferred into new beakers and im- mediately placed back in their designated pho- toperiod regimens. The number of cercariae in the original beaker was determined with the aid of a dissecting microscope by direct counts or by averaging ten 0.1 ml aliquots taken from a uniform cercarial suspension that was then adjusted to the number of cercariae/40 ml. The latter technique was necessary when cer- carial emergence was copious in a sampling interval. Cercarial release in the six 2 h light periods and six 2 h dark periods were separately pooled and compared with a Mann-Whitney test to determine if the median cercarial re- lease for the two species was significantly dif- ferent in the split 24 h photoperiods. In the 12L:12D and 12D:12L analyses, the light time interval consisted of 11.5 h of light and 0.5 h of dark, while the dark interval consisted of 11.5 h of dark and 0.5 h of light due to the times selected to evaluate cercarial emer- gence. A Kruskal-Wallis test was used to de- termine if the timing of the mean peak for cercarial release during a 24 h period varied significantly among the years 1990, 1992, and 1995 for E. trivolvis and C. vesicaudus. A probability of P < 0.05 was considered signif- icant for all statistical tests. Means are report- ed with associated standard errors. RESULTS A significant difference (Mann-Whitney test; P = 0.0233) was found in the median cercarial release for E. trivolvis between the light and dark intervals. An average of 156.7 + 65.5 and 5.0 + 2.1 cercariae were released in the light and dark periods, respectively. Similarly, a significant difference (Mann-Whit- ney test; P=0.0233) was found in the mean cercarial release for this species in the invert- ed photoperiod. An average of 5.0 + 2.1 and 272.7 + 41.3 cercariae were released in the dark and light time intervals, respectively. Maximum emergence occurred at 1430 and 0030; little or no emergence was observed during the 2 h intervals of the dark periods (Figures la and 1b). Continuous emergence was apparent under conditions of constant light (Figure lc; range = 12-187 cercariae/2 h interval) and dark (Figure 1d; range = 7— 130 cercariae/2h interval). A significant difference (Mann-Whitney test: P = .0001) was found for the median cercarial release of C. vesicaudus between the light and dark intervals. An average of 153.8 + 57.5 and 2021 + 349.7 cercariae was re- leased in the light and dark periods, respec- tively. A significant difference (Mann-Whitney test: P = .0003) was also found for the median cercarial release in the inverted photoperiod. An average of 1713.6 + 249.5 and 52.07 + 135.9 emerged in the dark and light time in- tervals, respectively. Peak emergence was ob- Cercarial Emergence—Rosen et al. 25 AVERAGE # CERCARIAE RELEASED / SNAIL Figure 1. Effect of light on the mean number + SE of Echinostoma trivolvis cercariae released from the snail Helisoma trivolvis at 2 h intervals over 24 h at 26° C under the following photoperiods: (a) 12 h light (0700-1859): 12 h dark (1900-0659), (b) 12 h dark (0700-1859):12 h light (1900-0659), (c) 24 h light, and (d) 24 h dark. An average of five infected snails was used for each photoperiod. served at 2230 and 1230 and little or no emer- gence occurred during the 2 h intervals of the light periods (Figures 2a and 2b). Continuous . cercarial emergence was observed under con- ditions of constant light (Figure 2c; range = 499-2298 cercariae/2 h interval) and dark (Figure 2d; range = 177-808 cercariae/2 h in- terval). The long-term study showed that the mean daily time for peak cercarial emergence was significantly different for E. trivolvis (Kruskal- Wallis test: P = .0002), but not for C. vesi- caudus (Kruskal-Wallis test: P=.9482). Peak — — < Zz Nn i= ie Nn < f — i] 4 <9] < — pe < iS) -4 & 1S) * io) o < 4 i) > < Figure 2. Effect of light on the mean number + SE of Cephalogonimus vesicaudus cercariae released from the snail Helisoma trivolvis at 2 h intervals over 24 h at 26° C under the following photoperiods: (a) 12 h light (0700- 1859):12 h dark (1900-0659), (b) 12 h dark (0700-1859): 12 h light (1900-0659), (c) 24 h light, and (d) 24 h dark. An average of 15 infected snails was used for each pho- toperiod. emergence time for E. trivolvis shifted to 2230 (dark period) in 1995 from 1430 (light period) in 1990 and 1992 (Figure 3). By com- parison, the peak emergence time of C. vesi- caudus remained fixed at 2230 (dark period) over the same time (Figure 4). DISCUSSION Light significantly triggered the release of E. trivolvis cercariae and inhibited the emer- gence of C. vesicaudus cercariae in our 1990 and 1992 studies. Photoperiod inversion was 26 Transactions of the Kentucky Academy of Science 58(1) AVERAGE # CERCARIAE RELEASED / SNAIL TIME Figure 3. Comparison of the effect of light on the mean number + SE of Echinostoma trivolvis cercariae released from the snail Helisoma trivolvis in 1990, 1992, and 1995. Naturally infected snails were collected in June or July from Owsley Fork Reservoir, Kentucky, and assessed at 4 h intervals over 24 h at 26° C under a 12 h light (0700- 1859):12 h dark (1900-0659) photoperiod. An average of 10 infected snails was used for each summer/experiment. accompanied by a complete reversal in the timing of peak cercarial release for both spe- cies. This suggested that cercarial emergence of the species is subject to control by the ex- ogenous photoperiod under conditions of con- stant temperature. By contrast, Schmidt and Fried (1996) found that light vs. dark did not affect the intensity of E. trivolvis cercarial emergence. Their study provided no acclima- tion period for snails to the tested conditions (i.e., snails were maintained in the dark at 12° C and then placed in conditions of light or dark at 28-29° C for an immediate 1 hour evaluation of cercarial emergence). The dif- AVERAGE # CERCARIAE RELEASED / SNAIL TIME Figure 4. Comparison of the effect of light on the mean number + SE of Cephalogonimus vesicaudus cercariae released from the snail Helisoma trivolvis in 1990, 1992, and 1995. Naturally infected snails were collected in June or July from Owsley Fork Reservoir, Kentucky, and as- sessed at 4 h intervals over 24 h at 26° C under a 12 h light (0700—1859):12 h dark (1900-0659) photoperiod. An average of 10 infected snails was used for each summer/ experiment. ference between our results and those of Schmidt and Fried (1996) may be more at- tributable to experimental design than to pos- sible strain differences of E. trivolvis. Wagenbach and Alldredge (1974) found that cercariae of the digenean Plagiorchis mi- cracanthos, which normally emerge during the dark phase of a 12 h light:12 h dark cycle, show a rhythmic emergence in continuous dark. They indicated that this latter phenom- enon provided evidence for an innate emer- gence rhythm, “which is synchronized to pho- toperiod by the inhibition of light” (Wagen- Cercarial Emergence—Rosen et al. 7 bach and Alldredge 1974). Such an “entrain- ment” of the endogenous rhythm to light:dark cycles has also been suggested for emergence of Proterometra macrostoma cercariae, which normally emerge during the dark phase of a light:dark cycle but show peaks of emergence under conditions of continuous light (Lewis, Welsford, and Uglem 1989). The lack of such a distinct rhythm under conditions of contin- uous light or dark in our study suggests that no circadian rhythm exists for E. trivolvis or C. vesicaudus in H. trivolvis, but longer term studies (i.e., in excess of 24 h) will be required to clarify this. No change was observed in timing of peak cercarial emergence of C. vesicaudus in the 1990, 1992, and 1995 studies, but a significant shift was noted for E. trivolvis. Cephalogoni- mus vesicaudus is an autogenic species that completes its life cycle as an adult in the small intestine of the spiny softshell turtle Trionyx spiniferus, which is present at Owsley Fork Reservoir. The timing of peak cercarial release is apparently a stable biological characteristic for the population of this species at this locale. By comparison, mammals (e.g., muskrats) and a variety of migratory waterfowl (e.g., Cana- dian geese, mallards, etc.) present at Owsley Fork likely serve as definitive hosts for E. tri- volvis in this habitat. The migratory nature of the avian hosts provides an opportunity for pe- riodic introduction of new strains of E. trivol- vis into the existing Owsley Fork population of this species. This may be indirectly assessed by observation of new cercarial release pat- terns appearing in the population over time. In a somewhat similar study, Theron (1984) documented chronobiological variation (i.e., one strain experienced an earlier daily cercar- ial release than the second strain) in two Schistosoma mansoni populations from the same area, but different ecological foci. He linked this variation to the amount of murine host participation in the life cycle. Further ex- periments determined that the cercarial emer- gence pattern of S. mansoni has a genetic basis that is a consequence of selective pressures exerted by different host species (Theron and Combes 1988). It has also been noted that the response of Schistosoma japonicum cercariae to light varies with the geographical strain of the species (Gumble et al. 1957). Both daily and seasonal differences in cercarial emer- gence have been recorded for eight strains of P. macrostoma (Riley and Uglem 1995). De- velopment of these strains has been linked to the species composition of sunfish definitive hosts in a specific geographical region (Riley and Uglem 1995). In the present study, since both potential mammalian and waterfowl hosts of E. trivolvis exist in the same “ecolog- ical focus,” the resulting emergence patterns for this species at Owsley Fork likely represent blending of patterns from several different strains. The changing prevalence of these dif- ferent strains over time might explain the vari- ation observed in the long-term emergence patterns for E. trivolvis cercariae at the res- ervoir. Molecular techniques recently used to differentiate and establish relationships of spe- cies in the genus Echinostoma (Morgan and Blair 1995; Petrie, Burg, and Cain 1996; Sloos et al. 1995) may be utilized in future studies to establish genetic differences between these proposed strains of E. trivolvis. ACKNOWLEDGMENTS This study was supported by a grant from Merck & Company, the Andrew Mellon Foun- dation Trust, and the Appalachian College As- sociation to R. Rosen and the Department of Biology at Berea College. We acknowledge previous Berea College student participants who contributed to this study including Mar- ichelle Asuncion, Melissa Edlin, Jose Iagan, Kiely Law, and Manuel San. LITERATURE CITED Asch, H.L. 1972. Rhythmic emergence of Schistosoma mansoni cercariae from Biomphalaria glabrata: control by illumination. Exp. Parasitol. 31:350-355. Giovannola, A. 1936. Inversion in the periodicity of emis- sion of cercariae from their snail hosts by reversal of light and darkness. J. Parasitol. 22:292-295. Glaudel, R.J., and FJ. Etges. 1973. The effect of pho- toperiod inversion upon Schistosoma mansoni cercarial emergence from Biomphalaria glabrata. Int. J. Parasi- tol. 3:619-622. Gumble, A., Y. Otori, L.S. Ritchie, and G.W. Hunter. 1957. The effect of light, temperature and pH on the emergence of Schistosoma japonicum cercariae from Oncomelania nosophora. Trans. Am. Microsc. Soc. 76: 87-92. Lewis, M.C., I.G. Welsford, and G.L. Uglem. 1989. Cer- carial emergence of Proterometra macrostoma and P. edneyi (Digenea: Azygiidae): contrasting responses to light:dark cycling. Parasitology 99:215-223. Luttermoser, G.W. 1955. Studies on the chemotherapy of 28 Transactions of the Kentucky Academy of Science 58(1) experimental schistosomiasis HI. Harvest of Schistoso- ma mansoni cercariae by forced nocturnal emergence from Australorbis glabratus. J. Parasitol. 41:201—208. Morgan, J.A.T., and D. Blair. 1995. Nuclear rDNA ITS sequence variation in the trematode genus Echinosto- ma: an aid to establishing relationships within the 37-collar-spine group. Parasitology 111:609-615. Oliver, L. 1951. The influence of light on the emergence of Schistosomatium douthitti cercariae from their snail host. J. Parasitol. 37:201-204. Petrie, J.L., E.F. Burg IH, and G.D. Cain. 1996. Molec- ular characterization of Echinostoma caproni and E. paraensei by random amplification of polymorphic DNA (RAPD) analysis. J. Parasitol. 82:360-362. Riley, M.W., and G.L. Uglem. 1995. Proterometra ma- crostoma (Digenea: Azygiidae): variations in cercarial morphology and physiology. Parasitology 110:429-436. Rosen, R.B., J.M. Ilagan, J.S. Law, M. Asuncion, M.E. Denton, and M.L. San. 1994. Seasonal prevalence of three species of digenetic trematodes in the snail Hel- isoma trivolvis at Owsley Fork Reservoir, Kentucky. Trans. Kentucky Acad. Sci. 55:32-35. Schmidt, K.A., and B. Fried. 1996. Emergence of cer- cariae of Echinostoma trivolvis from Helisoma trivolvis under different conditions. J. Parasitol. 82:674-676. Sloos, B., J. Meece, M. Romano, and P. Nollen. 1995. The genetic relationships between Echinostoma cap- roni, E. paraensei, and E. trivolvis as determined by electrophoresis. J. Helminthol. 69:243-246. Smyth, J.D., and D.W. Halton. 1983. The physiology of trematodes. 2nd ed. Cambridge University Press, Cam- bridge, U.K. Theron, A. 1984. Early and late shedding patterns of Schistosoma mansoni cercariae: ecological significance in transmission to human and murine hosts. J. Parasitol. 70:652-655. Theron, A., and C. Combes. 1988. Genetic analysis of cercarial emergence rhythms of Schistosoma mansoni. Behav. Genet. 18:201—209. Wagenbach G.E., and A.L. Alldredge. 1974. Effect of light on the emergence pattern of Plagiorchis micra- canthos cercariae from Stagnicola exilis. J. Parasitol. 60: 782-785. Trans. Ky. Acad. Sci. 58(1):29-32. 1997. Classifying Free Bieberbach Groups Raymond F. Tennant Department of Mathematics, Statistics, and Computer Science, Eastern Kentucky University, Richmond, KY 40475 ABSTRACT Let F be a free group on n letters and G be a finite group. To each epimorphism €: F — G is associated a free Bieberbach group F/N’ where N is the kernel of the epimorphism € and N’ is the commutator subgroup of N. In low dimensions, the free Bieberbach groups are described. In high dimensions, the free Bieberbach groups are described for the case when G is cyclic or dihedral. A theorem is given describing a method for factoring certain free Bieberbach groups as a semi-direct product of a lower dimensional free Bieberbach group and the integral group ring ZG. INTRODUCTION Let G be a finite group. A group S is called a crystallographic group (on the point group G) if S contains a finitely generated maximal abelian torsionfree subgroup A of finite index so that S/A=G and G acts faithfully by con- jugation on A. The rank of A is called the di- mension of the crystallographic group. A tor- sionfree crystallographic group is called a Bie- berbach group (Charlap’s definition [1986, p.74]). The dimension of the Bieberbach group is given by Schrier’s formula n=|G|(mk(F)—1)+1, where F is the free group on k generators with k=rnk(A) (Lyndon and Schupp 1977, p.16). It is well known that B is an n-dimensional Bieberbach group if and only if R’/B is an n- dimensional flat manifold. Let (X | R) be a presentation of G with finitely many genera- tors, that is, G=F/N where F is the free group on X and N is the normal closure of R in F. This may be expressed in terms of the short exaciasequence, 1 —> Nv" h > G —> Ihlet N’=[N,N] be the commutator subgroup of N, N=N/N’, and F=F/N’. The sequence above induces a short exact sequence on N and F, 0 > N->F->G>2Q 1. The action by conju- gation of G on N gives N a ZG-module struc- ture. N is referred to as the relation module of F/N (Linnell 1981). F is known to be a Bie- berbach group and will be referred to as a free Bieberbach group (Farkas’ definition [1981)). It is noted that of the flat manifolds associated to these free Bieberbach groups, some are or- ientable and some are non-orientable. 29 Two free Bieberbach groups F, and F, are isomorphic if there exists a ZG-module iso- morphism a and a group isomorphism B such that the following diagram commutes: 0>5N,>F,>°GQO>1 ene = (eu dh 0>N,>°F,>G > 1 where N, and N, are the relation modules as- sociated to F, and F, respectively. Since N is a ZG-module with free abelian rank equal to n, it is convenient to write N as Z when we are viewing F as an actual crys- tallographic group. Further, Aut(N)=GL(n,Z) so the action of G on N induces a represen- tation @: G > GL(n,Z). Since F=Z"XG (as sets), then we may define multiplication on F by (n,g)(m,h)=(n+9(g)m+f(g,h),gh), where n,meZ" (thought of as column vectors), gheG, and feH*(G,Z"). The 2-cocycle f: GXG — Z* may be constructed from i(f(g,h))=s(g)s(h)s(gh)~!, where s: G > F is a normalized set map and i: Z" > F is the in- clusion map. It is known (Tennant and Turmer 1992) that F is torsionfree. For the case, GA{1}, this is equivalent to the 2-cocycle f: GXG ~ Z being nontrivial. For a given dimension n > 0, there is a free Bieberbach group isomorphic to Z" where the point group is G={1}. Note that in this case, the compact flat manifold associated to the free Bieberbach group Z" is the n-torus, the orbit space R'/Z”. 30 Transactions of the Kentucky Academy of Science 58(1) FREE BIEBERBACH GROUPS IN LOW DIMENSIONS Let G be a finite group. In dimension n = 1, a free Bieberbach group (other than Z with trivial point group) must have the point group G=Z,. The compact flat manifold associated to this group is the circle. In dimension n = 2, a free Bieberbach group must have point group G={1} and mk(F)=2 so F=Z?. There- fore, the first case of a Bieberbach group B that is not a free Bieberbach group is defined by the sequence 0 — Z? > B > Z, > 1 where R’/B is homeomorphic to the Klein bottle. In dimension n = 3, a free Bieberbach group (other than Z? with trivial point group) must have G=Z, and mk(F)=2. A result of Linnell (1981) states that if two epimorphisms €,; F, ~ G and e,: F, ~ G, with mk(F,)=rnk(F,), are Nielsen equivalent, then their ee ed free Bieberbach groups are isomorphic. Since any two epimorphisms €,, €,: F(a,b) > Z, are Nielsen equivalent, con- sider the epimorphism derived from the pre- >. b). This pre- sentation and the action of Z, on Z° induces a monomorphism g: Z, > GL(3,Z). Since det(g(a))=—1, the flat manifold associated to this free Bieberbach group is non-orientable. The crystallographers refer to this particular manifold (crystal) as belonging to the mono- clinic class (Brown et al. 1978). In dimension n = 4, a free Bieberbach group (other than Z* with trivial point group) must have point group G=Z, and mk(F)=2 Since there is only one Nielsen equivalence class of epimorphisms €: F(a,b) > Z;, there is only one free Bieberbach group with point group G=Z,. Since 2 does not divide |G|=3, the induced monomorphism ¢: Z,; > GL(4,Z) must satisfy det(g(g))=1 for all geG and so it follows that the flat manifold R‘/F is orientable. SOME FREE BIEBERBACH GROUPS IN HIGH DIMENSIONS Let k be a positive integer. If k is odd, there is one free Bieberbach group of dimension n=k+ | where the point group is G=Z,. The associated flat manifold is orientable by the same reasoning as above. Suppose k is even, G is a point group with |G|=k, and €: F(a,b) > G is an epimorphism. If G=Z, or G=D,, the dihedral group with k elements and g: G > GL(k+1,Z) is the monomorphism induced by the action of G on Z" then there exists geG with ord(g)=2 such that det(g(g))=—1. It fol- lows that the associated flat manifold is non- orientable. For the point group, G=Z,, all epi- morphisms, €: F(a,b) > Z, are Nielsen equiv- alent so all free Bieberbach groups with point group G=Z, are isomorphic. On the other hand, there exist k such that not all epimorph- isms €: F(a,b) > D, are Nielsen equivalent, so we may not apply Linnell’s result to conclude that there is only one (k+1)-dimensional free Bieberbach group with point group G=D,. Let e: F, > G be an epimorphism where F, is free on k generators. Suppose another generator is added to F, and mapped to | under € to form a new epimorphism €*: F,,,, > G. The follow- ing theorem describes how the respective free Bieberbach groups are related. Theorem Let €: F, > G and e*: F,,, > G be epi- morphisms such that € * (eel ), Sisk and €*(x,,,)=1, then F* is isomorphic to Fx,ZG where F and F* are the free Bicherback groups associated to € and €* respectively and x, represents a semi-direct product. Proof Recall that F may be expressed in terms of the short exact sequence jo N > he coat The action by conjugation of G on N induces a monomorphism ¢: G > Aut(N). Let s: G > F be a normalized set map and define a 2-co- cycle f: GXG > N by i(f(g,h))= s(g)s (h)s(gh)~1. By viewing F as setwise equal to NXG we may define multiplication on F by (i,g)(m,h)= (n+ (g)m+f(g,h), gh). A theorem of Lyndon (1962) states that N*=N/ZG as ZG-modules. The isomorphism a: N* —> N/ZG induces an isomorphism A: Aut(N*) — Aut(N/ZG) and the projection p: N/ZG —> N induces a pro- jection P: Aut(N/ZG) > Aut(N) so that the action of G on N* induces a monomorphism o*: G — Aut(N*) that satisfies P(A(p*(g))). The free Bieberbach group may be expressed in terms of the short exact sequence Free Bieberbach Groups—Tennant 31 () CSI Sree esar A set map s*: G — F* can be chosen so that the 2-cocycle f*: GXxG — N* defined by i*(f*(g,h))=s*(g)s*(h)s*(gh)~! satisfies p(a(f* (g,h)))=f(g,h). Define the epimorphism é : N* > N by €(n*)=p(a(n*)). Then 0 — ZG > N* > N —> 1 is a short exact sequence of ZG-modules. The group F* is equal to N¥ XG (as sets) so define the epimorphism é: ha F by €(n*, g)=(E(n*),g). The ker(é)=ZG and we have the following commutative dia- gram: 0 0 dt AL 07> ZG>~ ZG 1 y L L 0 > N* 59 F*¥ 5G 1 y A L 0O2>N> Fo Gol t L L 1 It ] Define the normalized set map §: N > N* by §(n)=a—!(n/0) where 0 is the additive iden- tity in ZG. The set map §: F > F* defined by §(n,g)=(8(),g) is then a homomorphism and so it follows that the sequence 0 ZG > F* — F > 1 splits and F*=Fx,ZG. REMARK For the case where F*=Fx.ZG, if F has di- mension n then F* has dimension n+|G| and F* can be viewed as a free Bieberbach group which has a subgroup, F, that is also a free Bieberbach group. To crystallographers, F* be- longs to the class of superspace groups [Janner and Janssen 1979]. Equivalently, R'/F is an n- dimensional flat manifold sitting inside of the (n+|g|)-dimensional flat manifold R™*'C/F*. Corollary 1 If Epi(F,,,,G) contains only one Nielsen equivalence class and €: F, > G and €*: F,,, — G are epimorphisms then F*=Fx,ZG. As an illustration, the (k+1)-dimensional free Bieberbach group (k odd) with point group G=Z, can be factored as the semi-di- rect product F=Zx,Z(Z,). There are certain point groups with the fol- lowing property (known as the “swap” prop- erty [Tennant and Turner 1992)). Swap Property Given any two epimorphisms €,,€,: F, > G, there exists a finite sequence of epimorph- isms e€,,€, --, e, where e, F,,, > G for i odd and e,;: F, > G for i even, 1Six G be two epimorphisms. If G possesses the swap property then F,x,ZG=F.x,ZG where F, and F, are the free Bieberbach groups associated to €, and é€, respectively. Example The class of finitely generated abelian groups possesses the swap property (Tennant and Turner 1992). Since point groups must be finite, we are concerned only with the subclass of finite abelian groups. Corollary 3 If G is a finite abelian group and €,,¢€,: F, — G are two epimorphisms then F x.ZG= F.x.ZG where F, and F, are the free Bieber- bach groups associated to €, and €, respec- tively. When F,x,ZG=F’x.ZG is said to be one step stably equivalent to F,. LITERATURE CITED Brown, H.R., R. Bulow, J. Neubuser, H. Wondratschok, and H. Zassenhaus. 1978. Crystallographic groups of four-dimensional space. J. Wiley & Sons, New York, NY. Charlap, L.S. 1986. Bieberbach groups and flat mani- folds. Springer Verlag, New York, NY. Farkas, D.R. 1981. Crystallographic groups and their mathematics. Rocky Mt. J. Math. 11:511-551. Janner, A., and T. Janssen. 1979. Superspace groups. Physica A 99:47-—76. Linnell, P.A. 1981. Relation modules and augmentation ideals of finite groups. J. Pure Appl. Algebra 22:143—- 164. 32 Transactions of the Kentucky Academy of Science 58(1) Linnell, P.A. 1985. A cancellation theorem for lattices Lyndon, R.C., and P.E. Schupp. 1977. Combinatorial over an order. J. Lond. Math. Soc. IH. 31:450-456. group theory. Springer Verlag, New York, NY. Lyndon, R.C. 1962. Dependence and independence in Tennant, R.F., and E.C. Turner. 1992. The swap conjec- free groups, J. Reine Angew. Math. 210:148-173. ture. Rocky Mt. J. Math. 22:1083-1095. Trans. Ky. Acad. Sci. 58(1):33-34. 1997. DISTINGUISHED SCIENTIST AND OUTSTANDING TEACHER AWARDS 1996 Distinguished Scientist Award Dr. D. Allan Butterfield—Professor of Chemistry and Director of Center for Membrane Sciences, University of Kentucky, Lexington, Kentucky D. Allan Butterfield received the degree of bachelor of arts with highest distinction from the University of Maine in 1968. After a 3-year high-school teaching position in Zimbabwe (then Rhodesia) he began his doctoral work at Duke University where he received the Ph.D. in physical chemistry in 1974. After a l-year NIH _ post-doctoral fellowship in 1974, also at Duke, he joined the Department of Chemistry at the University of Kentucky in fall 1975 as an assistant professor. In 1983 he was promoted to the rank of professor. Since 1986 he has been the director of the University of Kentucky’s Center of Membrane Sciences. Dr. Butterfield has over 140 publications in refereed journals or monographs, an additional 15 publications either in press or submitted for publication, and about 80 published abstracts. He has presented papers at professional meetings throughout the United States and in England, Canada, and Australia. He serves as referee for some 20 journals and organizations. He has been the principal investigator or co-principal investigator of over 20 funded grants valued at nearly $13 million. Dr. Butterfield’s research in the field of membrane science as it pertains to Alzheimer’s disease is most impressive. His research allows for a_ unified understanding of the causes of Alzheimer’s disease, contributing to the eventual development of methods for either prevention of the disease or minimization of its effects. Dr. Butterfield has taught 18 different courses at both the undergraduate and graduate levels and has given invited seminars at universities and colleges. He has directed 17 master’s-level and 14 Ph.D.-level students; currently seven graduate students are working under his direction. He has guided 18 undergraduates in NSF sponsored summer research; eight of these students co-authored referreed scientific publications based on their research. Dr. Butterfield has been involved in the placement of about 75 students in graduate and professional positions. Dr. Butterfield has served on numerous university and department committees. He is a member of several professional organizations and honorary societies, e.g., American Chemical Society, Society of Sigma Xi, Phi Beta Kappa Honorary Society, and Phi Kappa Phi Honorary Society. He serves on the editorial board of Journal of Membrane Science. Dr. Butterfield has also been involved with several community organizations, including Habitat for Humanity. It is obvious that D. Allan Butterfield’s research 33 program and teaching are having an immediate positive impact on intellectual growth in the Commonwealth. This in itself is a great accomplishment. However, it is also quite likely that his research, which is providing scientists with a better understanding of the underlying causes of Alzheimer's disease, will lead directly to improvement in the quality of life in the Commonwealth and throughout the world. Outstanding College/University Science Teacher Award Dr. Karan Kaul—Professor, Biology Principal Investigator and Program Area Coordinator, KSUCRS, Kentucky State University, Frankfort, Kentucky Karan Kaul earned the degree of bachelor of science in botany with honors in 1966 and the degree of master of science in botany in 1968 from the University of Delhi. He received a Ph.D. in biology from the University of Kentucky (UK) in 1972. After receiving his doctorate, he remained at UK as a postdoctoral fellow in the Depart- ment of Biochemistry until 1977. Dr. Kaul spent 3 years outside Kentucky in research-associate positions before beginning his career at Kentucky State University (KSU) in 1980. Early in his career at KSU most of his time was spent in botanical research. As time passed, however, he began to devote more and more time to teaching. He pro- gressed from assistant professor to associate professor in 1990 and earned the rank of full professor in 1993. Al- though Dr. Kaul is nationally known as a prominent re- searcher in botanical and biological studies (with 14 pub- lications since 1991), he has chosen to spend over half of his time in teaching cell biology, vertebrate embryology, histology, principles of biology, botany, and several other biology courses. Dr. Kaul spends many hours outside the classroom in mentoring students, in coordinating pre-health clinics, and in assisting students in national examinations, sometimes at his own expense. He cares about the education of his students, encouraging their learning through time spent inside and outside the classroom; for this, he has the ut- most respect of students and faculty members alike. Dr. Kaul was listed in Who's Who Among America’s Teachers this year. The Kentucky Academy of Science is proud to present to Dr. Karan Kaul the 1996 Outstanding University Science Teacher Award. Outstanding Secondary School Science Teacher Awards Douglas C. Jenkins—Instructor, Warren Central High School, Bowling Green, Kentucky 34 Transactions of the Kentucky Academy of Science 58(1) Douglas C. Jenkins is co-recipient of the 1996 Kentucky Academy of Science Outstanding Secondary School Sci- ence Teacher Award. Mr. Jenkins received the degree of bachelor of science in physics and mathematics from Western Kentucky University (WKU) in 1968. For the next few years, he worked as project engineer for the U.S. Army Night Vision Laboratory in Fort Belvoir, Virginia. During this time, in 1972, he received the Department of Army Civilian Outstanding Performance Award. In 1973 he returned to Kentucky where he worked while continu- ing his education. He began his career at Warren Central High School in 1976. In 1978, he completed requirements for the degree of master of arts in education from WKU; in 1986 he obtained his Rank I certification. During his spare time he became a part-time instructor in the Department of Physics and Astronomy at WKU and, more recently, a teacher for the Verbally and Math- ematically Precocious Youth Program. At Warren Central he has served as boy’s golf and tennis coach. He is an active member in many professional organizations and has served on multiple commissions and task forces. Mr. Jen- kins was one of the original planners of the Barren River Imagination Museum of Science, a hands-on science mu- seum that serves surrounding school systems. Through his leadership, most Warren Central High School physics stu- dents were involved in preparing and constructing exhibits in 1993. He continues to involve his students in maintain- ing exhibits and participating in museum programs. “Doctor J,” as Mr. Jenkins is known by students and faculty, has increased interest in science at Warren Central through innovative teaching practices. His teaching incor- porates innovative, motivational practices that create op- portunities for development of student interest and suc- cess. His instruction focuses on learning outcomes. His classes are active and emphasize cooperative and group learning using real-life experiences. Professional and cooperative, Mr. Jenkins certainly con- tributes in many ways to the teaching profession. He has long been recognized as an exceptional teacher. In 1983 he received the Presidential Award of Excellence in Sci- ence Teaching; in 1986, the Warren County Teacher of the Year Award; and in 1991, the Sallie Mae Teacher Trib- ute Award. The Kentucky Academy of Science is proud to present to Douglas C. Jenkins the Outstanding Secondary Science Teacher Award. He is an exemplary representative of the many fine teachers of our Commonwealth. Janet Yates Ward—Instructor, Reidland High School, Pad- ucah, Kentucky Janet Yates Ward is co-recipient of the 1996 Kentucky Academy of Science Outstanding Secondary School Sci- ence Teacher Award. Ms. Ward received from Murray State University the degree of bachelor of science in 1986, the degree of master of science in biology in 1988, and Rank I certification in 1996. She graduated summa cum laude from her class. In fall 1989 she began her career at Reidland High School. Since that time, she has earned an excellent reputation from students and faculty members alike. Ms. Ward has made a positive impact on the young people at Reidland. Demanding of her students and cre- ative in her teaching approach, she has a genuine love of science, a love reflected in her attitude. She is instrumen- tal in spurring her students to excellence; many have re- ceived awards for their work. As sponsor of the biology competition team, she was instrumental in her students’ accomplishments as exhibited by high finishes in compe- tition. One of her students was awarded a full-tuition scholarship to the University of Louisville, and another was invited to present a project at the Kentucky Junior Science and Humanities Symposium sponsored by the University of Louisville. Reidland Science Club, which is sponsored by Ms. Ward, won bronze and silver medals at the State Science Olympiad. Ms. Ward has been named in Who’s Who Among Amer- ican Teachers and has been active on the McCracken County Education Association and National Education Association. She is a member of the Alpha Chi National Honor Society and Kappa Delta Pi Honorary Education Society. In 1994 she was awarded the Golden Apple Achiever Award sponsored by Ashland Inc. Her enthusi- asm, her skill at innovation, and her genuine concern for biology education are exemplary. One of her students wrote, “Through Mrs. Ward’s biology class, I began to see that science was a great collection of discoveries. I feel I owe to her my utmost gratitude and appreciation for her instruction and ability to convey her love of science, which seems to be contagious!” We hope that many more stu- dents will catch the love of science from Janet Yates Ward. Trans. Ky. Acad. Sci. 58(1):35-36. 1997. NOTES Human Myiasis in Kentucky Caused by Cuterebra sp. (Diptera: Oestridae).—Species of Cuterebra (Dip- tera: Oestridae) parasitize rodents and rabbits. The rabbit Cuterebra can be found also in young dogs and cats (1, 2). Little is known about the host range of this genus in Kentucky. The few documented infestation rates for nat- ural hosts have been quite variable. Over 70% of the trapped specimens of Peromyscus leucopus, the white- footed mouse, on Kentucky Lake islands in western Ken- tucky and Tennessee were infested with Cuterebra fonti- nella fontinella (3). In another study, 36% of P. leucopus individuals captured in eastern Tennessee harbored the same oestrid (4). Only three adult specimens of this genus, all identified as C. fontinella, are in the entomology col- lection of the University of Kentucky. No biological infor- mation is associated with the material. The case of myiasis reported herein involved a 19-year- old male. The larva was submitted to the Cooperative Ex- tension Service for identification following its removal from a lesion on the ankle. I determined it to be a late second or early third instar Cuterebra sp. The larva was sent to Dr. R. D. Hall, University of Missouri, who con- firmed the identification (pers. comm.). Accurate identi- fication of immatures of this group is best accomplished with mature third instars so that cuticular spines and cau- dal spiracles can be examined (5). Most documented cases of human oestrid myiasis have involved the rabbit-infest- ing species (6) C. buccata and C. abdominalis. These par- asitize eastern cottontail rabbits and are the two members of this group ranging into Kentucky (R.D. Hall, pers. comm.). The man answered a set of questions for me that de- scribed the general circumstances of the incident at which the infestation might have started and its course. This in- formation is summarized as follows. The most obvious opportunities for exposure appear to have occurred on four occasions between 5 Sep and 10 Sep 1995 while the man was squirrel hunting in Greenup County, Kentucky. He wore long pants, long socks, and high-topped canvas tennis shoes on all trips. While hunt- ing, he sat at the base of trees in wooded areas for | to 2 hours at a time. In most cases he cleared away fallen leaves before sitting on the ground, but occasionally he sat on a fallen tree. The first indication of a problem was noticed on 14 Sep 1995. He experienced “ warm, itching sensation, similar to the feel of a mosquito bite” and discovered a small red bump on skin of the medial surface of an ankle. Two more bumps appeared 2 days later. At that point, the irritation was described as a “gnawing sensation as if the flesh were being torn.” Ten days after the initial bump appeared, there were five bumps spaced at about 2-cm intervals along the ankle. At that time, the largest bump was about 2 cm in diameter and protruded about 1.5 cm above the surface of the skin. Anxious about the increasing size and numbers of le- 35 sions, the man visited a physician on 25 Sep 1995. By that time, the largest bump had “come to a head like a boil.” The wound was diagnosed as a spider bite; topical appli- cation of a hydrocortisone cream was recommended for the area. After returning home, the man squeezed a single bot larva from the boil-like spot. The area itched for about a week after the larva was removed but healed normally. Following the anxiety associated with the removal of a live maggot from the boil, the young man in this case was relieved to learn the identification of the insect. He re- membered that he had occasionally encountered mature bot larvae in subdermal cysts or warbles when skinning squirrels taken on previous hunting trips. Eggs of Cuterebra spp. are laid near burrows or areas frequented by the host (7). Mature eggs can hatch rapidly in response to stimulation of the host, such as an increase in air temperature and CO, concentration (8). These mag- gots can complete a portion of their development in hu- mans (9). Human infestations usually occur when an in- dividual spends some time near a host lair and comes in contact with eggs. Some human subjects of oestrid myiasis are aware of a “stinging” feeling. This probably occurs as the larvae use their mouth hooks to enter the skin. Penner (10) observed the activity of three Cuterebra larvae intentionally placed on his skin. The stinging sensation developed within 9 to 17 minutes and as the larvae entered the skin. Complete penetration took at least 18 minutes. Edema and redden- ing of the area developed and persisted for 2 weeks even though the larvae were removed immediately after pen- etration. The site of infestation, the ankle, is unusual in this Ken- tucky case. Rice and Douglas (11) reviewed five previously reported cases and documented two new ones. Four were on the face in or around the nose or eyes. The remainder were on the neck or torso. In two of the cases, the indi- vidual was outdoors at the time and felt the initial pene- tration. Baird, Podgore, and Sabrosky (6) summarized the 30 cases of human infestation by Cuterebra known to 1982; Baird, Baird, and Sabrosky (5) provided an update 7 years later. All but four infestations were on the torso or head. The infestation site for three was unknown and one was on the scrotum. In this case from Kentucky, there was a delay of 4 days between the last hunting trip and skin penetration by the maggot. These trips were the most obvious times that this individual could have come in contact with Cuterebra ovi- position sites. However, a few cases indicate no history or activity that places the person near oviposition sites. In the Kentucky case, an egg may have been picked up and remained on a shoe until development was complete or it was stimulated to hatch. LITERATURE CITED. (1) Hall, M., and R. Wall. 1995. Myiasis of humans and domestic animals. Adv. Parasitol. (20C) 35:257-334. (2) Muller, G.H., R.W. Kirk, and D.W. Scott. 1989. Small animal dermatology. 4th ed. W.B. Saunders, Phil- 36 Transactions of the Kentucky Academy of Science 58(1) adelphia, PA. (3) Kollars, T.M., Jr. 1995. Factors affecting the distribution of bot flies (Diptera: Cuterebridae) on is- lands in Lake Barkley in Kentucky and Tennessee. J. En- tomol. Sci. 30:513-518. (4) Dunaway, P.B., J.A. Payne, L.L. Lewis, and J.O. Storey. 1967. Incidence and effects of Cuterebra in Peromyscus. J. Mammol. 48:38-51. (5) Baird, J.K., C.R. Baird, and C.W. Sabrosky. 1989. North American cuterebrid myiasis. J. Am. Acad. Dermatol. 21: 763-772. (6) Baird, C.R., J.K. Podgore, and C.W. Sabros- ky. 1982. Cuterebra myiasis in humans: six new case re- ports from the United States with a summary of known cases (Diptera: Cuterebridae). J. Med. Entomol. 3:263— 267. (7) Catts, E.P. 1982. Biology of the New World bot flies: Cuterebridae. Ann. Rev. Entomol. 27:313-338. (8) Catts, E.P. 1967. Biology of a California rodent bot fly Cuterebra latifrons Coquillett. J. Med. Entomol. 4:87— 101. (9) James, M.T. 1947. The flies that cause myiasis in man. USDA Misc. Publ. 631. (10) Penner, L.R. 1958. Concerning a rabbit cuterebrid, the larvae of which may penetrate the human skin (Diptera, Cuterebridae). J. Kan- sas Entomol. Soc. 67-71. (11) Rice, P.L., and G.W. Doug- las. 1972. Myiasis in man caused by Cuterebra (Diptera: Cuterebridae). Ann. Entomol. Soc. Am. 65:514-516.— Lee H. Townsend, Department of Entomology, Univer- sity of Kentucky, Lexington, KY 40546-0091. Trans. Ky. Acad. Sci. 58(1):37-54. 1997. Abstracts of Some Papers Presented at the 1996 Annual Meeting of the Kentucky Academy of Science AGRICULTURAL SCIENCES Geographic differentiation and allozyme variation in pawpaw (Asimina triloba, Annonaceae). HONGWEN HUANG#* and DESMOND R. LAYNE, Land-Grant Pro- gram, Atwood Research Facility, Kentucky State Univer- sity, Frankfort, KY 40601. The pawpaw (Asimina triloba) is the largest tree fruit native to the United States and the only temperate mem- ber of the tropical custard apple family. In 1994, Kentucky State University was established as the USDA-ARS-Na- tional Clonal Germplasm Repository for Asimina spp. Seedling trees from 400 pawpaw accessions representing 70 distinct geographic regions from 17 states are currently being grown at our research farm. In a preliminary study, 32 pawpaw cultivars or advanced selections were assayed in 30 enzyme systems, using an isoelectric focusing poly- acrylamide slab gel system of pH 4-9. Twelve enzymes produced high resolution without tissue specificity and were further used for evaluation of allozyme diversity of geographic populations. We discuss the degree of genetic diversity within populations and differentiation between populations as evaluated by the expected heterozygosity (He), the proportion of polymorphic loci (P), the average number of alleles per locus (A), Chi-squared analysis of allele frequency heterogeneity, Nei’s standard genetic dis- tance (D), and identity (1). Dendrograms were generated by cluster analysis using the unweighted pair group meth- od to demonstrate the relationships of geographic popu- lations in the 17 states evaluated. The strategy for germ- plasm conservation and cultivar development through breeding is discussed. Issues related to establishment of a “core” collection are addressed. Influence of shade and root-zone modification on early growth and development of pawpaw (Asimina triloba, An- nonaceae) seedlings grown in the greenhouse. DES- MOND R. LAYNE, Land-Grant Program, Atwood Re- search Facility, Kentucky State University, Frankfort, KY 40601. This experiment was designed to determine the optimal light level for growing pawpaw (Asimina triloba) seedlings in the greenhouse. In addition, we wanted to determine if modifying the root-zone would impact seedling growth and development. We examined the individual and com- bined effects of shade and root-zone modification on early growth and development of seedlings. Experimental treat- ments were imposed from sowing until the plants were destructively harvested. The experimental design was a split-plot where blocking was done by position in the greenhouse. The main plot of the experiment was shade. This was accomplished by growing seedlings under a wooden frame covered with shade cloth to reduce inci- 37 dent light intensity received by the plant by 30, 55, 80, or 95%. The control treatment was 0% shade or ambient greenhouse light level. The split plot was root-zone mod- ification. Half of all growing containers were untreated (control) while the other half were painted with Spin- Out™, a commercially available product that stimulates development of a finer root system. There were 40 rep- licate seedlings per experimental treatment combination per block. Seedling shoot length and unfolded leaf num- ber was recorded 2/week from seedling emergence until destructive harvest. At harvest, leaf samples were taken for chlorophyll determination. Whole plant leaf area was also determined. Leaves, stems, and tap and lateral roots were separated and dried to determine biomass parti- tioned to the respective organs. Based on the results of this study, optimal greenhouse growing conditions for pawpaw seedlings have now been refined and are dis- cussed. Pawpaw (Asimina triloba, Annonaceae) morphological development during seed germination and seedling emer- gence. C.L.H. FINNESETH# and DESMOND R. LAYNE, Land-Grant Program, Atwood Research Facility, Kentucky State University, 40601; R.L. Geneve, Depart- ment of Horticulture and Landscape Architecture, Uni- versity of Kentucky, Lexington, KY 40546. Stratified pawpaw (Asimina triloba) seeds were germi- nated in vermiculite at 25°C to study embryo develop- ment. Seedlings were destructively harvested every 3 days prior to radicle protrusion (day 12) and at 5-day intervals thereafter. At each harvest date, 10 seedlings were ran- domly chosen for length measurement (mm) and fresh and dry weight (mg) determinations. Initial length of the underdeveloped embryo was less than 2 mm, but by day 70 seedling length increased to more than 350 mm. Twelve days after sowing, simultaneous development of the radicle and cotyledons produced lengths of 3.4 and 3.0 mm, respectively. Neither hypocotyl nor epicotyl was visible at this time. At radicle protrusion, radicle, cotyle- don, and hypocotyl lengths were 4.4, 4.0, and 3.2 mm, respectively. Endosperm was the greatest proportion of dry weight (99.1%); radicle, cotyledons, and hypocotyl represented the remainder. The first aboveground indi- cation of germination was hypocotyl hook emergence, which occurred at 27 days. At this time, average seedling measurements (mm length/% dry weight) were 41.7/17.0, 16.7/5.7, and 11.9/1.2 for the radicle, hypocotyl, and cot- yledons, respectively. Endosperm dry weight had de- creased to 76.1% by this time. Cotyledons reached maxi- mum length at 40 days. At day 45, the development of a discernible epicotyl occurred and the percentage dry weights for the radicle, hypocotyl, cotyledons, epicotyl, and endosperm were 44.4, 12.7, 3.5, 0.4, and 39.0, re- 38 Transactions of the Kentucky Academy of Science 58(1) spectively. At day 50, the seedcoat containing the cotyle- dons and residual endosperm abscised; average radicle, hypocotyl, and epicotyl measurements were 182.0/93.8, 16.0/6.0, and 7.3/0.2, respectively. Using flood control reservoirs for paddlefish (Polyodon spathula) production. R. ONDERS,* S. D. MIMS, and C. WANG, Community Research Service, Kentucky State University, Frankfort, KY 40601. Kentucky has a large number of flood-control reservoirs that may be used for paddlefish (Polyodon spathula; Po- lyodontidae) production. Juvenile paddlefish stocked into these reservoirs will feed primarily on zooplankton natu- rally present in the water. Paddlefish grow rapidly up to 4 kg per year with no feed cost and little management. Paddlefish can be harvested within 2 years for its quality white meat, which is boneless and firm in texture. Poten- tially this production system also will permit paddlefish to reach sexual maturity and the female fish can be harvested for its roe, which is processed into high value caviar. Roe production will be greatly increased by implementing all- female production technology currently being developed at the Aquaculture Research Center, Kentucky State Uni- versity. Both the meat and roe of paddlefish will provide an alternative source of income for the landowners, com- mercial fisherman, and food processors in Kentucky. BOTANY & MICROBIOLOGY Cryopreservation of seeds of endangered Ohio wetland plants. LARRY A. GIESMANN,* Department of Biolog- ical Sciences, Northern Kentucky University, Highland Heights, KY 41099; VALERIE C. PENCE and NYREE CONARD, Center for Research of Endangered Wildlife, Cincinnati Zoo and Botanical Garden, 3400 Vine Street, Cincinnati, OH 45220. Cryopreservation of seeds in liquid nitrogen (LN,) has been shown to be an effective way of preserving germ- plasm of endangered plant species. With funding from the Robert H. Reakirt Foundation and the L and L Nippert Foundation, the Plant Division of the CREW facility at the Cincinnati Zoo and Botanical Garden has been ex- ploring the feasibility of cryopreserving seeds of endan- gered Ohio wetland plants. Most wetland species have or- thodox seeds and should be candidates for cryopreserva- tion. Seeds of 14 species from nine families of flowering plants were collected from several Ohio wetland sites and tested in the CREW labs. All species are listed by the Ohio Department of Natural Resources as endangered, threatened, or potentially threatened in the state. In our study, four treatments were used for each species with at least 20 seeds in each treatment. In all treatments, seeds were desiccated over silica gel for 7 d. Two groups of seeds were then exposed to LN, for 1 hr and rehydrated for 1 d. One group was planted in pots and placed in the greenhouse immediately; seeds in the other group were stratified at 5°C for 3 mo before planting. Seeds in the remaining two treatments were not exposed to LN, and served as controls. The seeds of eight species germinated at rates ranging from 3% to 74%. Stratification increased the germination rate in most species. Work continues on 16 additional wetland species collected in the past year. Fine structure of Mycena leaiana (Tricholomataceae) basidiospores. DONALD G. RUCH* and MARK C. ROBBINS, Department of Biology, Ball State University, Muncie, IN 47306. The basidiospore wall of Mycena leaiana is composed of a thin, single, electron transparent layer, which exhibits some variations in thickness. No distinctive surface orna- mentation or germ pore is present. The protoplasm is sur- rounded by a typical membrane, which lacks distinct in- vaginations. Centrally located nonmembrane-bound lipid droplets comprise the bulk of the protoplasm. Spores are uninucleate. Mitochondria with a few well-delineated plate-like cristae are present. Endoplasmic reticulum is scant. Ribosomes occur regularly attached to the ER and outer mitochondrial membrane, as well as being densely packed throughout the cytoplasm. Various sized single membrane-bound vacuoles containing an electron dense material are present. Microbody-like organelles are readily observed. These microbodies are probably glyoxysomes, since malate synthase assays are positive indicating the presence of the glyoxylate cycle in the spore. Photomorphogenic control of flower scape elongation in dandelion (Taraxacum officinale; Asteraceae). MICH- ELLE JO LEATHERS and DAVID LOWELL ROBIN- SON,* Biology Department, Bellarmine College, Louis- ville, KY 40205. Dandelion (Taraxacum officinale) is a major weed of turfgrass. One reason for this weediness is its ability to produce a large number of seeds disseminated with the assistance of a parachute-like pappus. The flower stalk (scape) aids in this seed dispersal by carrying the seedhead far above the canopy. Elongation of the dandelion scape, therefore, is an important ecophysiological function. Three experimental approaches were taken to study the effect of light on dandelion scape growth. First, we ex- amined scape elongation at ca. 70 different turfgrass sites. Three measurements were taken at each location: the heights of the dandelion flower and seedhead scapes, and the height of the turfgrass surrounding the dandelion plants. We found a significant, positive correlation be- tween the height of the turf and that of the dandelion scapes. We hypothesized that the turf canopy altered the light microenvironment in such a way that it altered scape growth. Our second experiment was done with dandelion scape explants treated with various wavelengths of light. Light inhibited scape growth, with flower scapes being more sensitive than the seedhead scapes. In our third set of experiments we examined whole-plant scape elongation at different wavelengths. Far-red light treatment resulted in longer flower scapes than did treatment with red, blue, or clear filters, indicating a role for phytochrome. Seed- head scapes were less sensitive to light. It appears, there- Abstracts, 1996 Annual Meeting 39 fore, that dandelion scape elongation is influenced both by light microenvironment and stage of development. Taxonomic status of the varieties of Seneca snakeroot, Polygala senega (Polygalaceae). AMY E. TRAUTH* and ROBERT F. C. NACZI, Department of Biological Sci- ences, Northern Kentucky University, Highland Heights, KY 41099. Seneca snakeroot, Polygala senega (Polygalaceae), is an uncommon plant ranging throughout most of eastern North America. Two varieties have been described for this species: variety senega and variety latifolia. The taxonomic merit of these varieties is controversial. In general, both the reproductive and vegetative features of variety latifolia are much larger than those of variety senega. The goals of this research are (1) to more accurately define taxonomic differences between the varieties and (2) to test the hy- pothesis that the varieties senega and latifolia are separate species with morphologic differences not merely induced by ecology. Most of my work on the project has been in the laboratory and field. In the laboratory, univariate and multivariate statistical analysis of 13 vegetative and seven reproductive characters has been completed on 165 spec- imens from 11 herbaria. Among these morphologic char- acters are length of the shoot and inflorescence, length/ width ratio of the leaves, length of the wing and keel, and size of the seed. Field research involved the gathering of specimens of both varieties and habitat data, including soil samples. Preliminary results suggest significant differences between the measured morphologic characters. These re- sults suggest that the varieties of P. senega are better treat- ed as separate species. GEOGRAPHY Effects of weather systems on migraine headaches: a pilot study. L. MICHAEL TRAPASSO,* Department of Geography and Geology, Western Kentucky University, Bowling Green, KY 42101; KENNETH EMBRY, Embry Clinic, 1733 Campus Plaza Court, Bowling Green, KY 42101. Over a period of 9 months in 1992, 15 migraine head- ache sufferers in the Bowling Green, Kentucky, area were surveyed to document the onset, intensity, and duration of their migraine headache attacks. Eighty-two cases were analyzed with respect to the meteorological conditions of temperature, relative humidity, precipitation, and baro- metric pressure in effect during the migraine episodes. Graphic analysis consisted of superimposing the migraine attack, represented as a coded line drawn through the time period of the attack, upon the meteorological re- cording instrument charts. This analysis revealed promis- ing relationships between the simultaneous onset of head- aches from several subjects with a drop in barometric pressure, a rise in temperature, and a rise in relative hu- midity. Statistical analysis, however, failed to find these relationships to be significant. In general it was found that the absence of precipitation and the day of the week main- tained the best relationships with the onset of migraine headaches. HEALTH SCIENCES Differences in health and diet status: African-American vs. Caucasian rural southern elderly. MARTHA MAR- LETTE,* SUSAN TEMPLETON, and C. J. LEE, Hu- man Nutrition Research, Kentucky State University, Frankfort, KY 40601. We surveyed 1554 rural elderly in nine southern states. The sample was 84.2% female, 15.8% male, 37.7% Afri- can-American, and 61.7% Caucasian; average age was 72.7. Occurrences of selected diet-related diseases were examined using ANOVA (SPSS). Significant (p < .05) dif- ferences between the African-American and Caucasian groups were found in the prevalence of diabetes (22.8% vs. 12.9%), hypertension (53.0% vs. 44.1%), osteoporosis (1.8% vs. 10.2%), and atherosclerosis (2.5% vs. 6.1%) and cancer (1.6% vs. 4.7%). Two non-consecutive 24-hour food intakes were analyzed (Nutritionist IV) for average daily nutrient intakes. A Dietary Status Index (DSI), in- cluding Dietary Adequacy Status (DAS) and Dietary Mod- eration Status (DMS) factors, was computed; all dietary differences reported were significant at p < .05. African- American participants had a significantly lower DSI than Caucasian participants—a mean of 47.9 vs. 52.4. Though no significant difference for DAS was found, African- Americans did have lower intakes for vitamin E (111% vs. 136% RDA), calcium (65% vs. 73% RDA), and magne- sium (68% vs. 82% RDA). African-Americans had a sig- nificantly lower mean DMS (49.9 vs. 57.4); though their sodium intake (2100 vs. 2205 mg) was lower, African- Americans had significantly higher cholesterol intakes (327 vs. 228 mg), and more calories from fat (33.1% vs. 32.0%) and saturated fat (12.6% vs. 12.0%). These find- ings demonstrate the need for nutrition education for Af- rican-Americans, especially programs focusing on the re- lationship between fat intake and health problems. Health status and dietary knowledge and intakes of southern rural elderly. SUSAN TEMPLETON,* MAR- THA MARLETTE, and C. J. LEE, Human Nutrition Re- search, Kentucky State University, Frankfort, KY 40601. Rural elders (n = 1554) in nine southern states report- ed arthritis (63%), hypertension (47%), heart (26%), gas- trointestinal (18%), diabetes (17%), respiratory (13%), kidney (10%), osteoporosis (7%), atherosclerosis (5%), anemia (5%), and cancer (4%) problems. Smoking status, weight status, and chronic health problems were com- bined into health scores, range 9-100, mean 77.2; this correlated highly (r = .3832, p < .0001) with self-reported health status (“poor” to “excellent”). Surveys assessed re- spondents’ dietary knowledge; two non-consecutive 24-hour food intakes were analyzed for nutrient intakes (Nutritionist IV) and food group servings. ANOVA (SPSS) was used to determine significant (p < .05) differences among participants with low (LHS), medium (MHS), and high (HHS) health scores. The HHS group had signifi- 40 . Transactions of the Kentucky Academy of Science 58(1) cantly greater fiber knowledge (72.6% of responses cor- rect vs. 67.9%, LHS and 66.7%, MHS) and dietary fiber intake (15 grams daily vs. 13, LHS and MHS). The HHS group also had a significantly higher expectation about fruit servings required: 2.3 daily vs. 2.1 for both LHS and MHS. The HHS group consumed significantly more fruit (1.3 servings daily) than the MHS group (1.0) but not the LHS group (1.1); both MHS and HHS groups consumed significantly more grain products (3.9 and 4.1 servings dai- ly, respectively) than the LHS group (3.6). Fat and cho- lesterol knowledge, fat calorie intake, cholesterol intake, and sodium intake did not differ significantly among the LHS, MHS, and HHS groups. Participants with the best health appear to be those who recognize good fiber sources and include fiber-rich items in their diet. MATHEMATICS Polynomial conservation laws of the generalized Em- den-Fowler equation. B.D. VUJANOVIC, Faculty of Technical Sciences, University of Novi Sad, 21121 Novi Sad, Serbia; A.M. STRAUSS, Department of Mechanical Engineering, Vanderbilt University, Box 1612 Station B, Nashville, TN 37235; S. E. JONES, College of Engineer- ing, The University of Alabama, Tuscaloosa, AL 35487- 0276; PETER P. GILLIS,* Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506. We consider the polynomial conservation laws of fourth degrees with respect to x of the generalized Emden-Fow- ler equation x + (b/t) x + yx*t! = 0. We demonstrate that the existence of conservation laws depends upon the so- lution of a system of partial differential equations, usually termed the generalized Killing’s equations. The general form of the fourth degree conservation laws of the Em- den-Fowler equation is given; some concrete examples are discussed. Surface geometry of the Morehead radio telescope and the optimization of operating parameters. BRIAN M. LEWIS,* CHRISTOPHER L. SMITH,* and BENJA- MIN K. MALPHRUS, Morehead Astrophysical Obser- vatory, Morehead, KY 40351. The operating parameters of the Morehead Radio Tel- escope (MRT) at Morehead State University (Morehead, KY) affecting its performance are the half power band- width, the minimum detectable flux density, and the an- tenna gain. The MRT is in the process of being resurfaced to improve its radiation pattern and performance charac- teristics by optimizing the reflector geometry. The surface geometry has a direct effect on the radiation pattern, which determines the operating parameters. To determine these parameters, the precise surface area of the antenna is needed. The purpose of the current research is to de- termine the surface area and to compare the theoretical parameters before and after the telescope is resurfaced. The empirical and theoretical methods for approximation of the original surface and the new surface are discussed. The empirical method for the old surface is finite in na- ture and consists of the approximation and summation of planar areas. The empirical methods for finding the new surface is based upon the fact that a curved surface is an altered flat surface. Both theoretical methods involve in- tegral approximations and manipulations. Values from the empirical and theoretical surface areas of the old surfaces are compared to determine the improvements in perfor- mance characteristics. MOLECULAR & CELL BIOLOGY Alkylation of DNA by strain-induced electrophilic cyclic rings. ELIZABETH M. THOMAS* and ARTHUR CAM- MERS-GOODWIN, Department of Chemistry, Univer- sity of Kentucky, Lexington, KY 40502. Strain-induced electrophilic cyclic rings should alkylate the minor groove of the DNA fragments containing AAAG sites on the 5’ to 3’ strand. Alkylation of DNA would then inhibit DNA translation, thus proving to be a possible anti-tumor/anti-cancer drug agent. 3-phenylcyclobut-2-en- l-one and 2-propanone-3-(3-phenylcyclobuten-1-diyl) should exothermically alkylate DNA, releasing 2-3 kcal of ring strain in a first chemical step and potentially releasing approximately 27 kcal in a second irreversible step. The research includes the syntheses of these two compounds using zinc in an inert surrounding. DNA (15-mer) incu- bated with 3-phenylcyclobut-2-en-1-one showed no signs of alkylation by electrophoresis. This result needs to be checked by nmr. Alzheimer’s PS-1 mutation alters calcium homeostasis and sensitizes neurons to death induced by A and trophic factor withdrawal. BILL BARRETT,* QING GUO, and MARK MATTSON, Department of Anatomy and Neu- robiology, University of Kentucky, Lexington, KY 40502. Mutations in the presenilin-1 (PS-1) gene on chromo- some 14 are linked to autosomal dominant, early onset, Alzheimer’s disease. The amino acid sequence of PS-1 predicts an integral membrane protein with a structure suggesting functions as a receptor or ion channel, or in protein trafficking. PS-1 is localized to the endoplasmic reticulum (ER), and it is shown here that expression of a PS-1 mutation (L286V) in cultured neuronal cells exag- gerates Ca** responses to agonists (carbachol and brady- kinin) that induce Ca** release of the ER. Cells expressing L286V exhibit increased vulnerability to amyloid B-pep- tide toxicity and apoptosis induced by trophic factor with- drawal. The endangering action of L286V involves oxida- tive stress and disruption of calcium homeostasis, and an- tioxidants and calcium channel blockers counteract the ad- verse consequences of this PS-1 mutation. By perturbing Ca?* homeostasis, PS-1 mutations may sensitize neurons to age-related accumulation of AB and reduced trophic support. Assessment of metal impact in the Robinson Forest creek system using metalliothionein. KEVIN COMP- TON,* and CHRISTER HOGSTRAND, Department of Biology, University of Kentucky, Lexington, Ky 40502. Abstracts, 1996 Annual Meeting 41 Metallothionein (MT) was chosen and analyzed as a prospect for being a good bioindicator in the Robinson Forest creek system (RFCS), Breathitt County, KY. MT is a low-molecular-weight protein known to be induced by heavy metals such as Cd, Cu, Hg, and Zn, and to also positively correlate with hepatic concentrations of those metals. In the RFCS, recent mining has posed the prob- lem of possible stream contamination. It was, therefore, important to obtain raw data on stream conditions to assist in future monitoring efforts. All water metal concentra- tions at the six sites sampled were found to be below EPA published guidelines. Four species of fishes were sampled but one prevalent species, the stoneroller (SR) (Campo- stoma anomalum), was found to have the least amount of fluctuation in MT concentrations between members at each site. A significant variance (p < 0.05) was also found between two sites when SR MT concentrations were com- pared. The relative prevalence of the species combined with the lack in fluctuation of their MT concentrations between site members lead to the conclusion that they could be a good species from which MT could be sampled for metal exposure monitoring. Results on possible cor- relations between hepatic metal and MT concentrations of the species are forthcoming. If the concentrations are shown to positively correlate then it could be concluded that SR’s would be a good species from which to sample MT to use as a bioindicator to show increasing metal pol- lution in the RFCS. The presence of correlations could also possibly indicate a metal of most concern. Calcium homeostasis neurotoxicity hypothesis: possible evidence through L-type calcium channel density In hip- pocampal subregions. ALEXANDER COON,* ROSE- MARIE BOOZE, and DAVID WALLACE, Department of Anatomy and Neurobiology, University of Kentucky, Lexington, KY 40536. Alzheimer’s Disease (AD) has been diagnosed by mea- suring neurofibrillary tangle and senile plaque densities. In addition to these pathological markers, considerable neuronal cell death occurs in the brains of patients with AD. Considerable research has focused on determining the mechanism by which selective neuronal cell death oc- curs. An altered calcium homeostasis is believed to be an integral part of this cell death mechanism. L-type calcium channels are primarily responsible for the movement of Ca?* ions across the neuronal membrane. However, the relationship between channel density and the pathological diagnosis of AD has yet to be fully elucidated. To further examine this relationship we selected [*H]PN200-110, a calcium channel antagonist, to perform radioligand bind- ing experiments. First, rat brain homogenates were used to obtain the channel affinity and B,,,,(density). Second, in vitro receptor autoradiography was performed with [7H]PN200-110 on 25 human hippocampus samples (AD, n = 15; control, n = 10). Tissue was sectioned to 20 wm using a cryostat and then incubated with 1.0 nM [7H]PN200-110. Non-specific binding was binding in the presence of 1.0 1m Nifedipine. The autoradiograms were then analyzed with computer-aided densiometry to quan- tify the density of the channels in the hippocampal sub- regions. Preliminary data suggests that the binding of PN200-110 is specific to the L-type channels. However, the exact relationship between the channel density and the AD pathology is still unclear at this time. Characterization of AD-like pathology in normal aging. CHRISTOPHER R. BRACKNEY,* LARRY SPARKS, and STEVE SCHEFF, Department of Anatomy and Neu- robiology, University of Kentucky, Lexington, Ky 40536. Alzheimer’s disease is characterized by a progressive cognitive deterioration coupled with marked personality changes. The most notable pathological changes associat- ed with Alzheimer’s disease are Senile Plaques (SP’s) and Neurofibrillary Tangles (NFT’s). SP’s contain the B-amy- loid protein, a 39-42 amino acid peptide fragment derived from a transmembrane amyloid precursor protein (B- APP). NFT’s consist of a mass of introcellular argento- philic fibers, which are Paired Helical Filaments (PHF) resulting from abnormally phosphorylated tau protein. The present study assesses which B-APP peptide compro- mise the B-amyloid immunoreactivity in the brains of non- demented, non-heart disease, individuals 20-80 years of life. By using specific antibodies we will be able to con- clusively demonstrate which portion of the B-APP is ac- cumulating in normal aging. The accumulation of the C-100, a fragment derived from B-APP, precludes the formation and deposition of SP’s. The two immediate derivatives of the C-100 frag- ment are B-A42(43) and B-A40. It has been shown that B-A42(43) is found in greater accumulations then B-40 in Alzheimer’s disease. To examine the presence of B-A42(43) and B-A40 in normal aging we used specific antibodies to discrete regions of the B-APP peptide se- quence. We found several individuals who had the B-42(43) with lower densities then that of Alzheimer’s dis- ease. There were no accumulations of the B-40 in normal aging. It should also be noted there was no correlation between the presence of B-A42(43) and age. These results suggest that the B-APP processing in normal aging is sim- ilar to that found in Alzheimer’s disease. Chromosome 17 deletions in a subset of ovarian tu- mors. LYNN CARRICO* and MAURA PIERETTI, De- partment of Pathology and Laboratory Medicine, Univer- sity of Kentucky, Lexington KY 40506. Ongoing studies in the Pieretti laboratory indicate that different alterations in cancer-controlling genes occur in different histological types of ovarian tumors. The labo- ratory has already compiled data concerning genetic mu- tations on 145 tumors. However, in order to statistically correlate this genetic information to patients’ survival and recurrence, a larger number of cases needs to be evalu- ated. For this purpose, 45 additional cases were identified through the Kentucky Cancer Registry, which has a record of clinical, demographic, and follow-up data from Ken- tucky cancer cases after 1991. These tumors can be stud- 42 Transactions of the Kentucky Academy of Science 58(1) ied for the genetic alterations that characterize the differ- ent histological types: genetic deletions of chromosome 17, p53 mutation, and K-ras activation. Genetic deletions of chromosome 17 were studied by polymerase chain re- action of microsatellite markers and were identified as the loss or partial loss of one allelic band. The tumor tissue was removed from paraffin embedded blocks by a “punching out” method, and the DNA was phenol extracted and ethanol participated. The resultant DNA was then studied for deletions using the previously mentioned method. Preliminary results are available for 13 of the 46 total tumors. Five tumors showed no dele- tions on chromosome 17. Six tumors showed deletions on both arms of chromosome 17. Two tumors had deletions confined to the short arm of chromosome 17. All genetic deletions were observed in tumors of high grade. Further studies will be conducted with these samples and the re- sulting data will be evaluated for prognostic significance. Determination of cytokine production following OVA injection into the eyes of adoptive transfer mice. JASON E. DELONG,* JEROLD W. WOODWARD, RITA EGAN, and RICHARD BLACK, Department of Immu- nology and Microbiology, University of Kentucky, Lexing- ton, KY 40502. The eye, along with the central nervous system, has been regarded as an immune-privileged site where normal immune responses such as inflammation are suppressed and antibody production is favored. Anterior chamber-as- sociated immune deviation (ACAID) is a stereotypic sys- temic response to ocular antigens that involves dendritic cells assisting in trafficking antigens to the spleen where t-cells are activated and an immune response is elicited. The ACAID phenomenon is characterized by a shift in the T-helper 1/T-helper 2 balance of responses toward a dominant Th2 response. This alteration should be evident in the cytokine profiles of mice following immunization with antigen into the eye. Using an adoptive transfer mouse model with an immunoflourescent intracellular cy- tokine staining procedure, we have been able to detect and analyze individual cytokines secreted by antigen-spe- cific cells. The data attained through this project demon- strates successful isolation and recognition of individual IL-2 and IL-4 cytokines using the staining procedure with flow cytometry. This establishes the intracellular cytokine staining procedure as a useful tool in analyzing the im- mune response following antigen injection. Development of a sensing system for antimonite based on genetically engineered bacteria and green fluorescent protein. JANET MERCER,* SRIDHAR RAMANA- THAN, and SYLVIA DAUNERT, Department of Chem- istry, University of Kentucky, Lexington, KY 40506. A sensitive and selective sensing system for antimonite based on genetically engineered bacteria will be de- scribed. In this system, the selective binding of a bacterial protein to antimonite was coupled with fluorescence. When antimonite enters certain bacteria, it is effluxed out of the bacterial cell by a selective protein pump. The structural proteins ArsA, ArsB, and ArsC form the protein pump. A set of five genes, called the ars operon, code for the three structural proteins, in addition to two regulatory proteins, ArsR and ArsD. In the absence of antimonite, the ArsR binds to the promoter region of the ars operon inhibiting the expression of the protein pump. ArsR has a highly specific binding site for antimonite. Thus, when an- timonite reaches the cytoplasm it binds to ArsR inducing a conformational change, which releases the promoter re- gion of the ars operon. This induces the expression of the protein pump, which effluxes the antimonite out of the cell. Bacteria were genetically engineered to introduce the gene of Green Fluorescent Protein (GFP) downstream from the ArsR gene. The concentration of GFP in cells can be monitored easily by checking for fluorescence. Thus, the concentration of the GFP can be related to the concentration of the antimonite in the cell. Effect of retinioc acid on TIMP and stromelysin in HT 1080 human fibrosarcoma cancer cells. KEITH D. BRICKING* and RAYMOND E. RICHMOND, Depart- ment of Biological Sciences, Northern Kentucky Univer- sity, Nunn Drive, Highland Heights, Ky 41099. Recent research indicates that stromelysin, a matrix me- talloproteinase (MMP) expressed by a large percentage of malignant cancer cells, is capable of degrading extracel- lular matrix proteins. Matrix degradation is an important first step for cancer cell invasion, which is a precursor of metastasis. Stromelysin may be inhibited by a specific class of naturally occurring polypeptide inhibitors known as tissue inhibitors of metalloproteinases (TIMP). TIMP appears to act by non-covalently binding to stromelysin. Therefore any therapeutic agent affecting cellular expres- sion of stromelysin or TIMP may alter invasion. We ex- amined the hypothesis that retinoic acid (a vitamin A an- alog believed to have anticancer effects) will increase TIMP expression in a highly metastatic HT 1080 human fibrosarcoma cell line. These alterations will decrease the invasive properties of the cell. Stromelysin and TIMP ex- pression in 10-° M retinoic acid and vehicle control treat- ed cells were detected by polyacrylamide gel electropho- resis and immunoblot analysis. Invasion of HT 1080 can- cer cells after addition of retinoic acid and vehicle control has also been analyzed quantitatively using Matrigel In- vasion Chambers. Our preliminary data indicate that there was no change in stromelysin expression, an increase in TIMP expression, and a significant decrease in the rate of invasion after the addition of retinoic acid. These results suggest that future antimetastatic agents should focus on TIMP expression as well as matrix metalloproteinases. Effect on platelet-derived growth factor A-chain gene transcription by nm23 proteins in the WR-082-01 cell line. MATTHEW SINEX,* BIN LIU, JAMES REID, and DA- VID KAETZEL, Department of Pharmacology, Univer- sity of Kentucky Medical Center, Lexington, KY 40536. Platelet-derived growth factor (PDGF), a dimeric gly- Abstracts, 1996 Annual Meeting 43 coprotein comprised of two disulfide-linked polypeptide chains (A and B), is a powerful mitogen found in normal and malignant cells. This laboratory has previously re- ported that a 1 kb region of the PDGF A-chain promoter located between nucleotides — 1800 and —880 (relative to the transcription start site, +1) exhibits potent transcrip- tional repression. Within that negative regulatory region (NRR), a 31 bp silencer element (5’SHS) was identified at approximately — 1400. Screening of a cDNA expression library derived from HeLa cells with a probe derived from the 5’SHS sequence yielded clones encoding nm23, a pro- tein previously implicated in cancer metastasis. Subse- quent studies revealed that purified nm23 binds with high affinity to the 5’SHS sequence. The purpose of this study was to determine whether nm23 plays a role in mediating repression of the PDGF A-chain promoter. WR-082-01 cells, which are deficient in nm23 production, were co- transfected with DNA constructs directing overexpression of the H1 and H2 isoforms of nm23 and various A-chain promoter fragments. The NRR was inactive in these cells, suggesting that nm23 was important for its repression ac- tivity. Overexpression of nm23-H1 and H2 resulted in a general repression (< 2-fold) of all promoter constructs analyzed. While these nm23 isoforms exerted no addition- al inhibition upon vectors containing the 5’SHS element, an additional two-fold repression was conferred by H1 upon vectors containing the entire NRR. These data sug- gest that nm23-H1 mediates repression of A-chain gene transcription, probably via interactions with multiple si- lencer elements dispersed throughout the NRR. Effects of nucleotide excision repair mutants on mis- match repair. MARCI ADKINS* and ISABEL MEL- LON, Department of Pathology, University of Kentucky, Lexington, KY 40502. We have previously shown that mismatch repair mu- tants are also deficient in transcription coupled repair. These results are important because they imply a possible correlation between what was previously thought to be distinct DNA repair systems. Present work has been un- dertaken to further explore the connection between these two repair processes. Since microsatellite instability is a good marker for defective mismatch repair, we are spe- cifically looking for microsatellite instability in Xeroderma Pigmentosum (XP) cell lines. XP patients have deficiencies in one or more nucleotide excision repair genes, which have been named XP-A through XP-G. We have examined XP-A, which is involved in damage recognition, and XP-C, which repairs non-transcribed DNA. Clones of these cell lines have been examined using primers specific for cer- tain dinucleotide repeat sequences in a Polymerase Chain Reaction (PCR). The PCR products are then run on a 6% polyacrylamide gel and viewed by autoradiography. No microsatellite instability has been detected in either one of these cell lines. Therefore, our results deny a connec- tion between mismatch repair and transcription coupled repair. Electrophysiological studies of the SCN8a voltage-gated sodium channel. BRIAN DELISLE* and JONATHAN SATIN, Department of Physiology, University of Ken- tucky, Lexington, KY 40502. SCN8a is a voltage-gated sodium channel isoform ex- pressed in motor neurons. Loss of expression of the alpha subunit in transgenic mice results in Motor Endplate Dis- ease (MED). Transgenic mice often display a phenotype that mimics human disease. Symptoms of MED include progressive paralysis and juvenile death. The alpha sub- unit of voltage-gated sodium channels consists of four highly conserved domains. Most variability among the iso- forms occurs among the one-two linker of the alpha sub- units. This region contains functional glycosylation and protein kinase A phosphorylation sites. Utilizing RT-PCR protocol, we cloned the functional sites of SCN8a. Chi- meric channels are currently under construction. The pro- cess involves ligating digested BR2a, an extensively stud- ied isoform, and cloned SCN8a. The chimeras are ex- pressed in Xenopus laevis oocytes. We use the 2-electrode voltage clamp to study the kinetic properties of the ex- pressed ion channels and express the BR2a alpha subunit with and without the beta-1 subunit. Coexpression of the beta-1 subunit with BR2a modulates the rate of current decay and shifts voltage dependance of the channels. All experiments were then performed in the presence of 8-bromo-cAMP, promoting the phosphorylation of the so- dium channel. The alpha subunit plus and minus the beta-1 subunit both demonstrate a modest change in cur- rent amplitude. We use these experiments to validate a system to study the chimeric channels. The study is being used to extend how coexpression of the beta-1 subunit further modifies the sodium channel's kinetics. Identification of cytokines responsible for enhancing neonatal immune responses to polysaccharides. NIKOLE L. GILBERT,* RALPH CHEL VARAJAN, and SUB- BARAO BONDADA, Department of Microbiology and Immunology, University of Kentucky, Lexington, KY 40502. Polysaccharide antigens elicit a protective immune re- sponse in adults but not in neonatal individuals, thereby making neonates especially vulnerable to pathogenic, polysaccharide-bearing bacteria such as Streptococcus pneumoniae. Past research has shown that the B cell an- tibody response, which mediates immune protection in adults, is dependent on the presence of certain cytokines. It was thus hypothesized that the neonatal B cells may be competent to respond to polysaccharide antigens, but fail to do so due to a deficiency of cytokines required to stim- ulate neonatal B cells. To obtain an adult-like B cell re- sponse, murine neonatal splenocytes were cultured in vitro for 4 days with the polysaccharide antigen TNP-Fi- coll and various cytokines known to effect B cell activity. The plaque-forming cell-assay technique was used to mea- sure the number of antibody forming cells per culture. Neonatal splenocytes cultured with IL-4 and IL-5 togeth- er and IL-1, IL-5, and IL-6 separately resulted in 40-70% 44 Transactions of the Kentucky Academy of Science 58(1) of the adult response, while individual cytokines IL-2, IL-3, IL-4, IL-10, and GM-CSF provided no enhance- ment of the neonatal immune response. Neonatal splen- ocytes cultured with a combination of cytokines IL-1 and IL-5 produced 139% of the adult response to TNP-Ficoll. In conclusion, our studies have shown that the neonatal B cells can respond to polysaccharides as well as adult B cells if an adequate mixture of cytokines is provided, sup- porting the concept that the poor response of neonates to polysaccharides is likely due to a deficiency in cytokine availability. Identification of genes involved in splicing. ELIZA- BETH OTTE* and BRIAN RYMOND, Department of Biology, University of Kentucky, Lexington, KY 40506. The removal of introns from mRNA precursor, a pro- cess vital to eukaryotes, is carried out by the spliceosome. This complex enzyme consists of five small nuclear RNA molecules and 50-100 protein subunits. A genetic screen hoping to identify one or more of these spliceosomal pro- teins was performed. In this screen, a mutant intron was imbedded within the lacZ gene to produce a yeast strain in which production of the enzyme B-galactosidase was dependent upon the proper removal of this intron. Since this fusion gene’s intron was poorly excised, this strain was pale blue when grown in the presence of the chromogenic substrate, X-gal. My project was to characterize a set of recombinant plasmids which, when present in 20-50 cop- ies per cell, turned the colonies a much darker blue, ap- parently having enhanced B-galactosidase expression. It was hypothesized that such genes might suppress the in- hibitory effects of the intron mutation and increase pre- mRNA splicing efficiency. The ratio of pre-mRNA to ma- ture mRNA and the level of B-galactosidase in each pu- tative suppressor was assayed. The data obtained indicate that, under the conditions of assay, none of the suppres- sors enhanced splicing efficiency or enzyme production. I conclude that the enhanced blueness of these strains is due to either an indirect effect, such as enhanced per- meability of the X-gal substrate, or to enhancement of lacZ transcription, pre-mRNA splicing, mRNA or protein stability, or enzyme activity when cells are grown on the surface of an agar plate but not when grown (as assayed) in liquid culture. Innervation of immune tissues and the role of nerve growth factor. MARK PARRISH* and SONIA CARL- SON, Department of Anatomy and Neurobiology, Uni- versity of Kentucky Medical School, Lexington, KY 40536. The existence of a structural and functional relationship between the nervous and immune systems has been elu- cidated in detail over the last decade. Immune tissues are innervated by the sympathetic nervous system and the re- lease of neurotransmitter modulates a variety of immune responses. This modulatory relationship is well docu- mented, but the plethora of directional cues guiding these neurons to their immune tissue targets is not well under- stood. Increasing evidence suggests that a protein, Nerve Growth Factor (NGF), plays a significant role in directing the pattern of peripheral innervation resulting from the dependence of neurons on a target derived supply of this factor. Recent work with a line of transgenic mice, altered to overexpress NGF in skin, has given evidence that this growth factor may also be influential in guiding sympa- thetic innervation to immune tissues. NGF transgenics show dramatic changes in density and pattern of inner- vation to spleen and peripheral lymph nodes. To deter- mine if spleens from transgenic mice release a substance that alters neurite outgrowth, sympathetic ganglia were co-cultured with spleen from transgenic and non-trans- genic animals in the presence/absence of NGF and out- growth was measured. Cultured ganglia exhibited growth away from spleen which was reduced when cultured with transgenic tissue or in the presence of NGF suggesting that transgenic spleen retains NGF capable of stimulating outgrowth. However, ganglia paired with transgenic spleen also demonstrated changes in neurite morphology and a reduced total neurite area which we believe may be the result of cytokine production. Localization of pyrimidine biosynthesis enzymes with fusion proteins. DANA R. WEAVER,* JIANYUAN LUO, and JOHN M. RAWLS, School of Biological Sciences, University of Kentucky, Lexington, KY 40502. De novo pyrimidine biosynthesis is carried out by six enzymes contained in three proteins in animals: the CAD protein, DHOdehase, and the UMP synthase protein. These proteins are encoded by the r. dhod. and r-l genes respectively in Drosophila. Biochemical cell fractionation studies have shown that the CAD and UMP synthase pro- teins are cytosolic, whereas DHOdehase is located in mi- tochondria. We are carrying out studies to determine the distributions of these proteins within cells and among dif- ferent tissues, using fusion proteins containing the Aequo- rea victoria green fluorescent protein (GFP) and the he- magglutinin epitope (HA). A series of recombinant con- structs was created containing the GFP and HA open reading frames fused to the open reading frame of the dhod gene. Transgenic animals were produced that con- tain those fusions and genetic complementation confirmed in vivo function of the transgenes. Patterns of expression of the transgene protein were studied by UV fluorescence microscopy (GFP) and immunocytochemistry (HA epi- tope) in cells known to abundantly express CAD and DHOdehase: ovaries (i.e., nurse cells) and testes (i.e., elongated spermatids). Results of these experiments will be presented. Microsatellite instability (MIN) and TGF-B Type II re- ceptor gene mutations in human pancreatic carcinoma. CINDY HARP,* CAROL SWIDERSKI, and JAMES FREEMAN, Department of Surgery, University of Ken- tucky Chandler Medical Center, Lexington, KY 40536. Tumor progression is a multi-step process involving an increase in positive growth signals and a decreased re- sponse to negative growth regulation. In pancreas cancer Abstracts, 1996 Annual Meeting 45 this process often involves the mutational activation of the ras cellular oncogene and a concomitant loss in function of tumor suppressor genes. We found that a common characteristic of pancreatic cancers is loss in negative growth regulation by TGF-. This loss in regulation is often mediated by lack of expression of the TGF-B type II receptor gene (RII), a reported tumor suppressor gene. Microsatellite instability (MIN) has been proposed as a mechanism that targets repetitive sequences in the RII gene for mutation, resulting in loss of RII expression and function. The purpose of this study was to determine the incidence of MIN in pancreas cancer and to determine whether MIN targets the RII gene for mutation. Six loci were analyzed for the occurrence of MIN in pancreas can- cer specimens obtained from 16 different patients. The results show that MIN (9 of 16, 56%) and loss of hetero- zygosity (LOH), (7 of 16, 44%) are common in pancreatic tumors. We further analyzed prospective and retrospec- tive pancreas tumor specimens by *?’-SSCP for RIT mu- tations at regions comprising 2 potential MIN target sites. Mutations at potential MIN sites were detected in only 3 of 21 (14%) specimens all of which were in 5’ region of the gene that included a polyadenine repetitive sequence. RII mRNA was not detectable in 5 of 12 (42%) prospec- tive samples. These studies suggest that there is a greater incidence of MIN in pancreas tumors than reported for other tumor types. MIN, LOH, and loss in RII expression may play a role in the pathology of this disease. However, MIN targeted RII mutations occur in only a small sub- population of pancreas tumors. Therefore mechanisms other than MIN must account for the lack of RII expres- sion found in many pancreas tumors. Neuroprotective effects of nicotine in MPTP-treated C57B1/6 mice. ALAN NORTHINGTON+# and JAMES R. PAULY, College of Pharmacy, University of Kentucky, Lexington, KY 40536. Parkinson’s disease (PD) is a neurodegenerative disor- der characterized by a selective loss of CNS dopamine- containing neurons. Epidemiological studies have consis- tently demonstrated an apparent neuroprotective effect of cigarette smoking on the development of PD, with smok- ers having approximately half the risk of nonsmokers of developing the disease. Mice injected with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) de- velop a syndrome resembling human PD with a loss of nigrostriatal dopaminergic neurons. The purpose of the present study was to examine whether nicotine pretreat- ment reduces the neurotoxic actions of MPTP on striatal dopaminergic neurons. Mice were injected with either sa- line or nicotine (1.0 mg/kg, sc.) 10 minutes prior to ad- ministration of MPTP (40 mg/kg, sc.) or saline. Locomo- tor activity, motor coordination, and body weight were monitored for 12 days following MPTP treatment. The brains from each treatment group were then used for the evaluation of [?H]-GBR12935 binding to the dopamine transporter and binding to neuronal nicotinic receptor subtypes (using [°H]-cytisine and [!*°I]-alpha bungarotox- in). Animals treated with saline and MPTP demonstrated significant behavioral toxicity and weight loss; these effects were clearly attenuated in animals pre-treated with nico- tine. In the striatum, MPTP caused a significant reduction in the binding of [*H]-GBR12935 and [°H]-cytisine but not [!*°I]-alpha bungarotoxin. The toxic effects of MPTP on [°H]-GBR12935 and [*H]-cytisine were diminished by nicotine pretreatment. Possible mechanisms of nicotine- induced neuroprotection will be discussed. Screening for mutations causing failures in the RNA sorting system in spermiogenesis. NATHAN NOVOTNY,+* JAN URSO, and JOHN RAWLS, School of Biological Sci- ences, University of Kentucky, Lexington, KY 40506. In spermatogenesis, post-meiotic differentiation of spermatids is an extensive morphogenetic program that takes place, largely or entirely, in the absence of transcrip- tion. It utilizes RNAs synthesized during spermatocyte growth, stored in an inactive form during meiosis, then translated at various times during spermiogenesis. Sper- matocytes have an active system that sorts RNAs to be used immediately from those to store for later use; the components of this RNA sorting system are largely un- known. The objective of this research is to identify com- ponents of this system among the numerous male-sterile mutations that have been characterized in Drosophila mel- anogaster, by seeking variant expression of dhod RNA in mutant flies. A spermatocyte-specific promoter element of the dhod gene leads to synthesis of RNA that, mediated by a specific 5' RNA sequence (dTCE), is sequestered for ultimate translation in elongated spermatids. In a primary survey, spermatogenesis stages in over eighty male-sterile strains have been examined for histochemical expression of the dhod product dihydroorotate dehydrogenase (DHOdehase). Most strains displayed essentially normal expression patterns for DHOdehase, however a few strains showed aberrant expression patterns that are pre- dicted for mutations of the spermatogenesis gene control system: zero or strongly reduced expression; expression of DHOdehase in early stages (possibly precocious expres- sion). As a secondary survey, mutants showing altered ex- pression of DHOdehase are being tested for expression of an array of lacZ reporter transgenes that contain various elements that control dhod expression: the dhod promoter or the promoter of another spermatocyte-expressed gene; with or without the dTCE sequence that controls trans- lational delay. Results of these experiments will be pre- sented. Site directed mutagenesis of a conserved phenylalanine in cytochrome P450 2E1. ERIC DEDRICK* and TODD PORTER, College of Pharmacy, University of Kentucky, Lexington, KY 40536. Cytochrome P450 2E1 is an enzyme that catalyzes the oxidation of a wide array of substrates, xenobiotics, and naturally occurring chemicals. The oxidative capabilities of P450s arise from a heme group folded deep within the molecule. Although it is not fully understood how an elec- 46 Transactions of the Kentucky Academy of Science 58(1) tron donated from its reductase partner is able to make the transition from the surface of the P450 molecule to the heme group in the interior, it is thought that a highly conserved phenylalanine at position 429 facilitates this transfer. Previous mutagenesis experiments have suggest- ed that hydrophobic compounds can substitute for this phenylalanine and still provide a functional molecule. Fur- thermore, because of its structure, proline may fit well at this position, causing a minimal distortion of the P450; notably, a proline has been discovered at this position in a functional P450 enzyme from flax seed. In this experi- ment, a proline codon was substituted for the phenylala- nine codon at this position using polymerase chain reac- tion. This mutation was then ligated into the expression vector pJL2/3a and expressed in Escherichia coli. Once positive mutants have been identified by sequencing, the protein will be characterized in spectrophotometric and catalytic assays. Stage dependent induced hatching in the muskellunge, Esox masquinongy. TAMARA SAPP,* SARAH M. BLANK, and JOHN J. JUST, Department of Biological Sciences, University of Kentucky, Lexington, KY 40506. The embryos of all aquatic animals are encased in pro- teinaceous egg cases from which they escape during de- velopment. Hatching often involves digestion of the egg case by a hatching enzyme secreted from hatching glands. This study shows that hypoxic environments can induce premature hatching in Esox masquinongy 9 days post-fer- tilization, 5 days before normal hatching. Embryos 9, 10, 11, and 12 days old were subjected to environments with various oxygen pressures by aerating vials containing em- bryos with oxygen (PO, = 760 mm Hg), air (PO, = 160 mm Hg), or nitrogen (PO, = 0 mm Hg) for 1 hour. At all stages tested, eggs aerated with nitrogen (n = 461) exhibited an increased hatch rate compared to the control embryos treated with air (n = 463), while those treated with oxygen (n = 461) showed a greatly reduced hatch rate. There was a progressive increase in percent hatch with those embryos treated with nitrogen. Percent hatch of 9, 10, and 11 day old embryos after 1 hour of nitrogen treatment was 46%, 90%, and 99% respectively, while 9, 10, and 11 day old control embryos treated with air had hatch rates of 0%, 1%, and 46%. It is believed that hatch- ing can occur only after the development of hatching glands and enzymes. Hatching medium was collected from a mass hatching of E. masquinongy embryos and the macromolecules (MW > 10,000) were concentrated via ultrafiltration. Protease activity was demonstrated by di- gestion of gelatin on photographic film. Work continues to isolate the hatching enzyme and correlate its develop- mental appearance with the ability to prematurely hatch. TNF-alpha neuroprotection in opiate-mediated astrog- lial toxicity. JENN JACKSON,* MARK P. MATTSON, and KURT F. HAUSER, Department of Anatomy and Neurobiology, University of Kentucky, Lexington, KY 40536. Opiate drugs of abuse liability, such as heroin and mor- phine, inhibit fetal brain development, in part by disrupt- ing astroglial mitosis and also by inducing astroglial hy- pertrophy. Although opioids affect growth, the mecha- nisms by which opioids disrupt neurobehavioral develop- ment are not understood. The mu-opioid receptor serves as a primary target of opiate drugs of abuse. In astroglia, mu-opioid receptor stimulation causes increases in intra- cellular calcium. The resulting elevations in intracellular calcium mediate morphine-induced alterations in astro- glial growth. The present study tested the hypothesis that a naturally occurring neuroprotective factor, tumor necro- sis factor-alpha (TNF-alpha), will prevent the deleterious effects of opiate drugs by blocking opiate-induced increas- es in intracellular calcium. Astrocytes were isolated asep- tically from the cerebral hemispheres of 1-day-old ICR mice. Astroglial cultures were pretreated with TNF-alpha at least 24 hours prior to the opioid insult to serve a neu- roprotective role. Astrocytes were subsequently treated with TNF-alpha in the presence or absence of morphine (1 2M). Intracellular calcium was analyzed by ratiometric analysis of the calcium indicator, fura-2. The study showed TNF-alpha, at concentrations of 10 ng/ml and 100 ng/ml, causes a concentration-dependent reduction in morphine- induced activation of intracellular calcium. These findings suggest that mu-opioid receptor and TNF-alpha signaling pathways converge and have opposing actions in calcium homeostasis. Studies in progress are assessing whether TNF-alpha attenuates morphine-induced alterations in as- troglial growth. Translational regulation of mRNA in human breast can- cer. BRAD SEGEBARTH* and STEPHEN ZIMMER, Department of Microbiology and Immunology, University of Kentucky, Lexington, KY 40502. This proposal is based on the hypothesis that increased initiation of messenger RNA (mRNA) translation in hu- man breast cancer cells is a fundamental mechanism me- diating increased tumor cell growth and the capacity to invade and metastasize. The eukaryotic initiation factor 4E (eIF-4E) is a 25 kilodalton (kDa) phosphoprotein involved in the recruitment of messenger RNA to the polysomes. This reaction is considered to be the rate-limiting step in the synthesis of cellular proteins, with eIF-4E as the least abundant subunit. Studies show that overexpression of eIF-4E in human breast cancer cells is associated with increased translation, cellular proliferation, transforma- tion, and tumorigenesis. PHAS-1 is a 20 kDa protein found to bind to eIF-4E. The binding seems to be regu- lated by phosphorylation, with the complex dissociating upon phosphorylation of PHAS-1. About 70% of mRNAs containing extensive 5’-region secondary structure are on- cogenes or important growth regulatory genes. One of these, basic fibroblast growth factor (bFGF), is a multi- functional molecule that affects cell growth and induces angiogenesis. Due to the extensive 5’ region secondary structure, bFGF, along with other malignant characteris- tics, may be disproportionally inhibited in cells overex- Abstracts, 1996 Annual Meeting 47 pressing PHAS-1. If this is found to be the case, potential medical utility stems from being able to effectively devel- op a drug that can affect the PHAS-1/eIF-4E interaction and suppress malignant properties in oncogenic cells. PHYSICS & ASTRONOMY Celestial seasons. RICHARD HACKNEY,* KAREN HACKNEY, ROGER SCOTT, CHARLES McGRUDER, MICHAEL CARINI, RICHARD GELDERMAN, SAN- DRA CLEMENTS, JOHNNY JENNINGS, JOHN NO- BLE, and RICO TYLER, Western Kentucky University Astrophysical Observatory, Department of Physics and As- tronomy, Western Kentucky University, Bowling Green, KY 42101. Many have only a casual acquaintance with (and an in- adequate understanding of the causal connection be- tween) the seasons and related observable astronomical phenomena. In our introductory astronomy courses, we attempt to engage students’ minds through personal ob- servation and interpretation of data to make the connec- tions and to see the astronomical reasons for the seasons. We describe a number of activities suitable for students at various levels to illustrate through student participation the primary astronomical observables related to the cycle of the seasons. We acknowledge funding by the Eisenhower Math and Science Program, the Kentucky PRISM-UG Program, and the NASA Kentucky Space Grant Consor- tium. Comet Hale-Bopp: Opportunity for education and de- veloping student involvement. ROGER SCOTT,* KAR- EN HACKNEY, RICHARD HACKNEY, CHARLES McGRUDER, MICHAEL CARINI, RICHARD GELD- ERMAN, SANDRA CLEMENTS, and JOHN NOBLE, Department of Physics and Astronomy, Western Kentucky University, Bowling Green, KY 42101. Comet Hale-Bopp, already visible with binoculars in the early evening sky, promises to be an easy naked-eye object in March and April 1997. Comets are notorious for being unpredictable in brightness. However, Hale-Bopp seems to be brightening about as expected and may put on a show even better than Hyakutake. Bright comets of- fer excellent opportunities for student projects and for ed- ucational outreach to the general public. They are easily photographed using any 35-mm camera capable of taking a time exposure. We discuss simple student projects and techniques for photographing the comet. Photographs of other bright comets of recent decades are shown as ex- amples. Charts and finding information are provided. Enigmatic world of extragalactic radio jets. CASEY WATSON,* ILDEFONSO GUILARAN, RYAN CULL- ER, MONICA DECKARD, RICHARD GELDERMAN, MICHAEL CARINI, JOHN NOBLE, SANDRA CLE- MENTS, ROGER SCOTT, KAREN HACKNEY, and RICHARD HACKNEY, Western Kentucky University As- trophysical Observatory and Center for Automated Space Science, Department of Physics and Astronomy, Western Kentucky University, Bowling Green, KY 42101. Radio galaxies, quasars, Seyferts, and blazars are all des- ignated by the term Active Galactic Nuclei or AGN. Among the most striking characteristics of AGN are the collimated radio jets, up to millions of light years in length, which emanate from their cores. The radio emis- sion is due to synchrotron radiation, which arises when charged particles from the active nucleus spiral around a magnetic field. However, the precise physical nature of the jets remains undetermined. Discovering the compo- sition of the jets is crucial because that insight will lead to greater understanding of the AGN’s central engine. It is the primary goal of this project to make a contribution to current jet theory. Our theoretical work will be com- plemented by observations, including images from the CCD camera at the WKU Astrophysical Observatory and the Hubble Space Telescope. We thank the NASA Ken- tucky Space Grant Consortium and the NASA Center for Automated Space Science for support. Gravitational lensing simulation for the classroom. MA- RIA FALBO-KENKEL+# and JOE LOHRE, Department of Physics and Geology, Northern Kentucky University, Highland Heights, KY 41099. Gravitational macro-lensing is a celestial phenomenon that can be observed when two or more objects at differ- ent distances from the earth lie along the same line of sight in the sky. The observation of this effect, which pro- duces distortions and apparent displacements of the ob- ject furthest from earth in the alignment, was listed by Einstein in his famous 1915 paper as one of three impor- tant tests of general relativity. There are ways to optically simulate gravitational lensing events. We present an in- expensive and simple method to make a lens that can be used to easily simulate gravitational lensing in the class- room. We discuss two effects that can be studied with the lens: (1) Einstein-ring formation and (2) multiple image formation. This work was supported by funds from the Kentucky Space Grant Consortium. Morehead radio telescope: design and fabrication of a research instrument for undergraduate faculty and student research in radio frequency astrophysics. BENJAMIN K. MALPHRUS,* BRIAN M. LEWIS, and CHRISTO- PHER L. SMITH, Morehead Astrophysical Observatory, Morehead, KY 40351. Faculty and students of the departments of Physical Sciences and Industrial Education and Technology at Morehead State University (MSU) (Morehead, KY) have designed and assembled the Morehead Radio Telescope (MRT) to provide a research instrument for undergradu- ate astronomy and physics students and an active labora- tory for MSU undergraduate students and faculty in phys- ics, engineering, and computer science and for under- graduate students, faculty, and science teachers through- out Kentucky. Goals of the MRT program are to enhance the curricula in physics, physical science, electronics, and 48 Transactions of the Kentucky Academy of Science 58(1) science education programs by serving to provide (1) a research instrument for investigations in astronomy and astrophysics; (2) an active laboratory in astronomy, phys- ics, electrical engineering, and computer science; and (3) a research instrument and laboratory for science teacher education and inservice programs. The telescope incor- porates a modular design in which components may be easily removed for use in laboratory investigations and for student research in design projects. The performance characteristics of the telescope allow a varied and in-depth scientific program. The sensitivity and versatility of the telescope design facilitate the investigation of a wide va- riety of astrophysically interesting phenomena. The MRT provides hands-on experience in research and instrumen- tation technology in a cutting-edge science, one that is in the midst of scientific revolution. This presentation pro- vides and overview of the MRT and its current operating status. NASA Kentucky Space Grant Consortium, Center for Automated Space Science, and EPSCoR opportunities for space-related science, technology, and education. KAREN HACKNEY,* RICHARD HACKNEY, ROGER SCOTT, CHARLES McGRUDER, MICHAEL CARINI, RICH- ARD GELDERMAN, JOHN NOBLE, SANDRA CLE- MENTS, and JOHNNY JENNINGS, Kentucky Space Grant Consortium, NASA EPSCoR Program, and Center for Automated Space Science, Department of Physics and Astronomy, Western Kentucky University, Bowling Green, KY 42101; MICHAEL BUSBY, Center for Automated Space Science, Center of Excellence in Information Sys- tems Engineering and Management, Tennessee State Uni- versity, Nashville, TN 37203. NASA has funded three programs in Kentucky to in- volve university faculty and students in space-related re- search, technology, and education. We describe (1) cur- rent opportunities in the Kentucky Space Grant Consor- tium for undergraduate scholarships and graduate fellow- ships for students in mentored, space-related projects; (2) funding opportunities for development of space-related research projects and activities to support teaching of space science and related disciplines; (3) progress of ex- isting projects and new opportunities in the NASA EPSCoR Program; and (4) special opportunities for mi- nority students in the Center for Automated Space Sci- ence. Use of computers for the enhancement of introductory astronomy for non-science majors. RAYMOND C. Mc- NEIL, Department of Physics and Geology, Northern Kentucky University, Highland Heights, KY 41099. Within the sciences, introductory laboratory experienc- es allow students a better understanding of scientific laws and a better appreciation of the scientific method leading to their discovery. At the heart of a laboratory course is the assumption that students will participate in “hands- on” activities that will provide experience in gathering data, analyzing the data to find relationships, and drawing conclusions from the experimental results. Traditionally, introductory astronomy students have been limited in that they generally cannot actively conduct experiments, con- trolling one or more of the variables, as can students in other sciences. The solution at Northern Kentucky Uni- versity (NKU) and other institutions has been to utilize laboratory computers to allow students to explore, then investigate in detail, simulations of astronomical phenom- ena. Observations may be repeated, and in some cases variables changed, just as in the laboratories of other sci- entific disciplines. The active nature of such experiences can enhance learning and instill a sense of the excitement of scientific discovery. The acquisition and initial use of a dedicated set of computers for the astronomy laboratory at NKU is described. Materials developed by the author or adapted from other sources are discussed, including Contemporary Laboratory Experiences in Astronomy, with copies available upon request. A list of software found to be most promising as the basis for laboratory activities or other types of student involvement is also in- cluded, as are future directions for this work. The author gratefully acknowledges the support of the National Sci- ence Foundation (Grant #9452219) and NKU. Optical monitoring of very high redshift quasars. SAN- DRA CLEMENTS, Western Kentucky University Astro- physical Observatory and Center for Automated Space Science, Department of Physics and Astronomy, Western Kentucky University, Bowling Green, KY 42101. The observations and results of a 1-year program to monitor high redshift quasars is reported. Quasars are among the most distant and energetic objects in the uni- verse. Believed to be caused by activity in the centers of galaxies, quasar behavior has been a challenge to under- stand. After their discovery, monitoring campaigns quickly noted variability in the radiation emitted by some quasars. Subsequent variability studies have uncovered a wealth of information about their underlying physics. However, due to a lack of monitoring data, such studies have yet to be performed on the most remote quasars, those with red- shift = greater than 4. Since first discovered in the late 1980s, about 50 of these quasars have been found. Com- paring the physical characteristics and behavior of quasars in the very early universe with less remote quasars will provide insight into their evolution. The variability studies required to determine the underlying physics of these ob- jects require monitoring data. To begin to satisfy this re- quirement, a program to monitor quasars with z greater than 4 was initiated in June 1995. to date, over a dozen objects have been monitored at the University of Florida's Rosemary Hill Observatory, Bronson, FL, and the West- ern Kentucky University Astrophysical Observatory, Bowl- ing Green, KY. Photometry with CCDs: Optimizing the signal extrac- tion. RYAN CULLER,* FONSIE GUILARAN, MONI- CA DECKARD, CASEY WATSON, MICHAEL CARI- NI, RICHARD GELDERMAN, JOHN NOBLE, RICH- Abstracts, 1996 Annual Meeting 49 ARD HACKNEY, KAREN HACKNEY, ROGER SCOTT, and SANDRA CLEMENTS, Western Kentucky University Astrophysical Observatory and Center for Au- tomated Space Science, Department of Physics and As- tronomy, Western Kentucky University, Bowling Green, KY 42101. The CCD detector has revolutionized astronomical photometry. Photometric precision better than 1% has been achieved for bright sources. Faint sources, such as Active Galactic Nuclei (AGN), can also be studied with photometric precision approaching 1%. This is a signifi- cant improvement over the 10% precision achieved with photographic plates and the 5% precision usually achieved with photomultiplier tubes. However, these improvements can be reached only through the proper processing of the CCD image and appropriate extraction of the brightness information from the processed image. We discuss (1) what we have found to be optimal methods for both the processing of CCD images and the extraction of bright- ness information and (2) application to the measurement of brightness variations in AGN. We acknowledge funding from the NASA Kentucky Space Grant Consortium and the NASA funded Center for Automated Space Science at Western Kentucky University. PHYSIOLOGY & BIOCHEMISTRY Effects of ovariectomy and dietary energy restriction on the body weight of Fischer 344 rats. C. BROWN,* Y. ZHANG, C. WANG, and C. J. LEE, Community Re- search Service, Kentucky State University, Frankfort, KY 40601. Thirty-two fischer 344 rats were randomly assigned into four groups of eight rats each. Group 1 was sham-oper- ated and fed the control diet (AIN-76A) ad libitum; Group 2 was ovariectomized and fed the control diet at the av- erage intake of Group 1; Group 3 were sham-operated and fed the energy-restricted diet at 60% of the average intake of Group 1; Group 4 were ovariectomized and fed the energy-restricted diet at 60% of the average intake of Group 1. Sixty grams of the energy-restricted diet provid- ed only 60% of the energy content, but equal amounts of protein, minerals, and vitamins of 100 g of the control diet. The experimental period was 8 weeks. There were no significant changes in the body weight of Group 1, whereas the body weight of Group 2 increased by 8% of the initial body weight. Both Group 3 and 4 lost ca. 20% of the initial body weight with no differences between the two groups. These results indicate that ovariectomized rats were more likely to gain weight than the sham-op- erated rats when fed the control diet and that both ovar- iectomized and sham-operated rats will lose weight when fed energy-restricted diets. Effects of ovariectomy surgery on urinary calcium ex- cretion of Fischer 344 rats. D. HAMPTON,* Y. ZHANG, C. WANG, and C. J. LEE, Community Research Service, Kentucky State University, Frankfort, KY 40601. Sixteen female Fischer 344 rats were fed AIN-76A for 1 week before they were placed in metabolic cages and the urine excretion was completely collected for 3 days. Half of the rats went through ovariectomy with the other half through sham-operation. One week after the opera- tion, rats were placed in metabolic cages with the urine collected for 3 days. Urine samples were analyzed for cal- cium by atomic absorption spectrophotometry. Dietary in- take, thus, dietary calcium intake, was lower after surgery than before surgery. Calcium intake post surgery was low- er for ovariectomized than for sham-operated rats. Uri- nary calcium excretion (mg/day) decreased by 20% for sham-operated rats and by only 5% for ovariectomized rats. These results suggest that surgery decreased calcium absorption from the gut, leading to lower urinary calcium excretion in sham-operated rats; increased calcium mo- bilization from the bone may be responsible for the higher urinary calcium excretion in ovariectomized rats. Gentamicin-induced nephrotoxicity in the teleost fish Astronotus ocellatus (Cichlidae). W. STEWART CURT- SINGER* and HONG Y. YAN, School of Biological Sci- ences, University of Kentucky, Lexington, KY 40506. Aminoglycoside antibiotics (AGs) are effective in treat- ing bacterial infections. The AGs also inflict ototoxic and nephrotoxic effects on treated subjects. Nephrotoxicity of gentamicin sulfate on fish was first described by Yan et al.; edema caused by gentamicin was documented subse- quently. We used histological and autoradiographical methods to document temporal course of intake of gen- tamicin as well as cellular/structural damages caused by the gentamicin to the fish kidney. The experimental sub- jects, the teleost fish Astronotus ocellatus, received one, two, three, or four daily intramuscular injection(s) of 20 mg/kg or 120 mg/kg gentamicin and sacrificed 1 day after the last injection. Fourteen fish also received one injection of *H-labeled gentamicin (5 Ci/10 gm fish). Two fish as a group were sampled 1, 2, 4, 8, 24, 36, 48 hours after the injection. Kidney tissues were dissected and processed with standard histological procedures. Kodak-NTB3 emul- sion was applied to sections with *H-labeled gentamicin. Exposure was carried out in a light-tight box up to 42 days and developed. Edema was obvious after just one injec- tion. The weight gain increased as fish received more in- jections in both dosage groups. There was no significant difference in weight gain between two treatments. An in- dication that 20 mg/kg dosage was sufficient to illicit ne- phrotoxic damages. The necrosis was evident in the prox- imal tubular epithelial cells. Autoradiographical results showed the maximal uptake of gentamicin occurred be- tween 36 and 48 hours after injection. The present study shows that fish kidney is a valid model for aminoglycoside nephrotoxicity study. Supported by Deafness Research Foundation, NIH-DC01729, and Howard Hughes Medi- cal Institute. Potential gentamicin-binding protein(s) in the ear of the cichlid fish Astronotus ocellatus (Cichlidae). STEVEN M. RIGDON,* W. STEWART CURTSINGER, BIN-TAO 50 Transactions of the Kentucky Academy of Science 58(1) PAN, and HONG Y. YAN, School of Biological Sciences and Department of Neurosurgery, University of Kentucky, Lexington, KY 40506. Evidence of aminoglycoside (e.g., gentamicin sulfate) damages to the auditory sensory hair cells of the teleost fish Astronotus ocellatus was previously demonstrated. Continuation of the study further showed regeneration of damaged ciliary bundles 10 days after maximal damage. Our preliminary results using both immunocytochemical and autoradiographical methods indicate that the incor- poration of gentamicin into hair cells occurs between 36 and 48 hours after intramuscular injection. To better un- derstand the ototoxic and regeneration mechanisms in- volved, Western blotting was used to identify the presence of potential gentamicin binding protein(s). The fish were injected with gentamicin sulfate (120 mg/kg) and sacri- ficed either 24 or 48 hours later. Control fish received only saline injection. Three tissues (ear, kidney, and brain) were dissected and processed following standard immuno blot- ting protocols. Results indicated the presence of potential gentamicin-binding protein(s) in ear and kidney tissues, except tissues from control animals and brain tissues. Higher degree of binding was observed in 48-hr tissues than 24-hr samples. These results corroborated earlier im- munocytochemical and autoradiographical findings on temporal uptake sequences of gentamicin. On-going ex- periments employing both biochemical and molecular techniques are designed to characterize the nature of the potential gentamicin-binding protein(s). It is hoped fur- ther understanding of gentamicin-binding protein(s) could lead to the development of prophylactic agent(s) against both ototoxic and nephrotoxic damages caused by ami- noglycoside antibiotics. Supported by Deafness Research Foundation, NIH-DC01729 and Howard Hughes Medical Institute. Relationship between muscle lipid content and body weight of paddlefish (Polyodon spathula). X. LOU, T. L. WHITE,* C. WANG, and S. D. MIMS, Community Re- search Service, Kentucky State University, Frankfort, KY 40601. Lipid content of fish meat is important to processing and marketing of fish meat. The objective of this study was to determine the muscle composition of paddlefish (Polyodon spathula: Polyodontidae) with various body weight. Twenty-five paddlefish (body weight 4 to 20 kg) were used for this study. Fillets with the red meat trimmed off were prepared and analyzed for moisture, total lipids, protein, and ash content. With increasing body weight, muscle moisture and protein content tended to decrease; muscle lipid content increased with ash content remaining constant. Muscle lipid content increased dras- tically when the body weight was 10 kg or more. These data will have practical implications for the harvesting of paddlefish and the ultimate use of paddlefish meat. Skeletal compartmentalization and metabolism of cal- cium in maturing male and female rats. D. L. DEMOSS* and G. L. WRIGHT, Physiology Department, Marshall University School of Medicine, Huntington, WV 25704. The *H-tetracycline (*H-T) bone-labeling procedure was employed to monitor bone resorption from urinary loss curves in male and female rats of various ages. In addition whole body dry skeletal mass and the loss of 3H-T from individual bones was determined. It was found that the dry skeletal mass/body mass ratio of females was sig- nificantly greater than for males, indicating the impor- tance of dry skeletal mass in quantitative assessments. The urinary loss of *H-T was described by a double exponen- tial equation (label loss from bone fluid and calcified skel- etal compartments). The results indicate the presence of two distinct and independent exchangeable bone com- partments as determined by the *H-T method. Both com- partments decrease in size with age but their label loss activities are different. The label loss activity within the rapidly depleted compartment, which we suggest affects calcium loss from the bone fluid compartment, is “inert” in the sense that it does not change with age or between sexes. Label loss activity in the slowly depleted compart- ment, which we suggest represents resorption of calcified bone, is increased with aging. Despite a smaller skeletal mass, resorptive activity in this compartment is higher in female rats than in male rats by 24 weeks of age due to a larger compartment size and a maintained higher rate of resorptive activity within the calcified compartment. Stability of paddlefish (Polyodon spathula) meat stored in crushed ice. X. LOU,* C. WANG, Y. XIONG,! G. LIU,! and S. D. MIMS, Community Research Service, Kentucky State University, Frankfort, Kentucky 40601; Department of Animal Sciences, University of Kentucky, Lexington, KY 40546. Fillets from six paddlefish (Polyodon spathula; Poly- odontidae) were cut into seven sections for each fish. The sections were stored in sealed plastic bags covered by crushed ice. One section from each fish was taken on day 0, 1, 2, 3, 5, 7, 10. The samples were analyzed for thio- barbituric acid-reactive substances (TBARS) as an indi- cator of lipid degradation and protein solubility as an in- dicator for protein degradation. The data were analyzed with the GLM procedure of SAS and the storage time, fish, and section were the independent variables. There were no significant differences among the sections. There were no significant changes in protein solubility or TBARS during the 10 days of storage. These results indicate that paddlefish meat stored in crushed ice is stable for at least 10 days. Producing the finest community college graduates. J.G. SHIBER, Division of Biological Sciences, Prestonsburg Community College, Prestonsburg, KY 41653. At Prestonsburg Community College. (Prestonsburg, KY) 998 students completed a questionnaire on how com- munity colleges (CC) can produce the “finest” graduates for the 21st century, i.e., people well prepared for the challenges ahead. Most said that the primary mission of a Abstracts, 1996 Annual Meeting 51 CC should be teaching and learning, and that its basic principle should be to provide a broad background so stu- dents can further their education and/or enter the job market. Successful graduates, they said, should be versa- tile, willing to pursue varied career options, and able to integrate disciplines. Nearly all agreed that to achieve this end, the CC should offer courses and programs reflecting basic economy and work force changes, draw upon all parts of society's talent base to train students, and reassess long-standing educational practices, especially in the sci- ences and engineering. Are these students’ expectations being met at their CC? When asked why enrollment is down and what should be done about it, or how the CC can best use its funds or improve its educational mission, the most frequent response was always “Provide more courses/degree programs/career options.” Many said that their CC did not offer all the courses their majors re- quired for entry into a university, that better facilities and student services (e.g., daycare and tutoring) were needed, and that more sensitivity to student needs and abilities, with less emphasis on non-academic matters, is essential. Of the 81% planning to attend a university, over half said the most important thing they needed to experience at the CC was improvement of personal discipline (study, note-taking, attendance habits). The results indicate that CC's may have to pay more attention to student academic needs and concems if they are to produce the “finest” graduates for the 21st century. Student attitudes toward the study of biology. JOHN R. MORRISON* and RUTH E. BEATTIE, T. H. Morgan School of Biological Sciences, University of Kentucky, Lexington, KY 40506. A pre- and post-course attitudinal survey was adminis- tered to students enrolled in freshman-level major and non-major biology courses at the University of Kentucky (Lexington, KY). The survey included questions on (1) the perceived importance of biology in everyday life; (2) the image of biology courses; and (3) factors that influenced the student to take a biology course. The surveys were self administered and the resulting data were analyzed using the T-test and mean values. All surveys were administered to students taught by the same instructor. Pre-course data analysis indicated that non-biology-major students had a less positive attitude than biology major students towards the importance of biology in everyday life. Post-course scores showed a significant increase in agreement with the statement that “Everyone should study biology” and that an understanding of biology is important in politics and aids intelligent voting. All test groups agreed strongly with the statement that “Biology involves a lot of memoriza- tion.” An interest in the subject matter was the primary factor in influencing students to take a biology course. The majority of students surveyed had a positive experience in their high school biology courses. Student perspectives of field investigations: misconcep- tions, problems, and educational benefits. TERRY L. DERTING* and JILL H. KRUPER, Department of Bi- ological Sciences, Murray State University, Murray, KY 42071. Life is the theme of biology, yet all too often it is stud- ied only in artificial laboratory environments. Despite the inherent appeal of the outdoors, the idea of conducting field investigations of biological phenomena is often met with trepidation. We conducted a case study of a field ecology class in which groups of students initiated and completed field investigations. Many of these students had no prior field experience. Throughout the course we mon- itored the feelings, problems, and successes of each group. Student misconceptions regarding the projects included the necessity of sophisticated knowledge and equipment and the inability of peers to contribute useful input to other groups projects. Problems encountered were diffi- culty focusing on a specific testable hypothesis, difficulty observing experimental subjects, lack of experimental con- trols, difficulty drawing conclusions from data, disagree- ments among group members, and time constraints. Groups whose data did not support their hypotheses, or whose data collection proceeded more slowly, viewed their project as less good than groups whose data supported their hypotheses or who gathered data more quickly. The educational benefits realized included recognition of the importance of flexibility and patience when conducting an investigation, awareness of biases in methodology and data presentation, advantages of working in a group, and rec- ognition of students’ abilities to pose and investigate eco- logical questions. The class consensus was that field in- vestigations are frustrating yet exciting and rewarding. The students agreed unanimously that continual monitoring of each student's progress, thoughts, and feelings was of great benefit. Traditional vs. computerized education: a student sur- vey. J.G. SHIBER, Division of Biological Sciences, Pres- tonsburg Community College, Prestonsburg, KY 41653. A survey of 873 students at Prestonsburg Community College (Prestonsburg, KY) reveals their opinions about learning via computer technology (including TV, videocas- settes, Internet) versus the traditional approach of attend- ing live lectures, taking notes, using textbooks, etc. The majority said that they prefer the traditional style of learn- ing and that computer learning is more expensive, less humane, and even less efficient. Most said that computers will be essential in their careers but believe that ultimate success in career and life is best derived from a traditional education, which they define as more well rounded than a computerized one. Teacher-student feedback, the ma- jority agreed, is necessary for quality learning, and pro- fessors lecturing in class can give more information, through elaboration, than computers. Most students large- ly attributed our present social problems and inability to solve them to the fast rate of our technological develop- ment. They were almost equally split, however, as to whether or not computer use would eventually cause the demise of society as we know it, or if we can live without 52 Transactions of the Kentucky Academy of Science 58(1) computers. The majority believed that humans are not organically (physically and mentally) evolved enough to be completely dependent on computers. Nor did they see exceptional, near-future benefits from computers. Indeed, the trend towards computerization of everything bothers the majority of those surveyed. Despite their preferences and reservations, however, most students seemed resigned to the inevitability that computerized education will even- tually replace the traditional. Undergraduate course in science ethics for biology and chemistry majors: I. development and content, FRANK H. WILBUR, Department of Biology, Asbury College, Wilmore, KY 40390. In 1986 selected ethical dilemmas relating to the health professions were incorporated into PHP 201, Introduction to the Health Professions, a course offered at Asbury Col- lege (Wilmore, KY) and open to any student interested in exploring career options in health-related fields. In 1990, PHP 201 was dropped in favor of requiring a one-credit- hour course in Science Ethics (BIO 372) for all senior biology and chemistry majors. This course, which is now required of all junior science majors, has been modified each of the 6 years it has been taught. An introduction to ethics and ethical theory, as well as discussions on world views, values, character development, and the prevailing cultural moral climate, precede class exercises focusing on solving ethical situations and dilemmas in scientific re- search, medicine, and ecology. While the ethical dilemmas presented have varied somewhat from year to year, those dealing with aspects of information disclosure (truth tell- ing and informed consent), allocation of health care, pro- fessional conduct in scientific research, and environmental stewardship have been regularly included. Course evalu- ations submitted by all students enrolled in the course have played a major role in course modifications. The most recent evaluations reveal considerable student sat- isfaction and enthusiasm for course format and content. Undergraduate course in science ethics for biology and chemistry majors: I. world view and values, FRANK H. WILBUR, Department of Biology, Asbury College, Wil- more, KY 40390. In 1990 a one-credit-hour course in Science Ethics (Bio 372) required for all biology and chemistry majors was added to the science curriculum at Asbury College (Wil- more, KY). It readily became apparent that many students taking the course lacked the foundational skills necessary to properly address ethical situations and dilemmas. They had little or no knowledge of basic ethical theory, had not sufficiently formulated their world view, and had not clar- ified their personal value system. The course, as currently offered, addresses these concerns. Twenty-five percent of the course is devoted to basic ethical theory and assess- ment of the present cultural climate. Students are pre- sented with examples of different world views. These may include a Biblical world view from the Christian and the Jewish perspective, a postmodern world view, a relativist world view, and/or a humanistic world view. Students are required (1) to formulate and articulate their own world view and to use it as a foundation in the ethical decision- making process, (2) to identify the sources of their moral beliefs, and (3) to articulate and defend their personal value system. Teaching objectives include consciousness- raising and sensitizing; and values analysis, clarifications, and criticism. Student response to this approach, as de- termined by comments and student evaluations, has been most positive. Also, the ability of students to more intel- ligently apply appropriate ethical decision-making pro- cesses to solving ethical situations and resolving ethical dilemmas has shown marked improvement. ZOOLOGY & ENTOMOLOGY Analyses of size and density of Tectarius muricatus (Lit- torinidae) on San Salvador Island, Bahamas. MELINDA L. CRAWFORD,* JACQUELINE M. GRANESE, and RUDOLPH PRINS, Department of Biology, Western Kentucky University, Bowling Green, KY 42101. Samples of Tectarius muricatus collected from transects at nine sites on San Salvador Island in the Bahamas were counted and measured with regard to vertical zonation on their respective rocky shores. Analyses were conducted using a two-way ANOVA with no interaction effects. Size differences were found among the different sites. How- ever, the experimental results for T. muricatus contradict those of many other littorinids in that no significant ver- tical size gradient existed in the populations. Densities of the snails within and between the sites were analyzed to determine if vertical gradients existed. No significant dif- ferences in density were found among the sites, but a sig- nificant differential was exhibited between the lowest two zones of the transects when averaged across all sites. These results are discussed with respect to several possi- ble environmental and physical factors that may have af- fected the size and density distributions of T. muricatus. Quantification of patterns such as distribution, density, and size are necessary in preliminary studies of this vir- tually unknown littorinid so that its superior physical tol- erance may be investigated. Characterization of snapping action of the alpheid shrimp Alpheus heterochaelis. JARED FIALKOW* and HONG Y. YAN, School of Biological Sciences, University of Kentucky, Lexington, KY 40506. Alpheus heterochaelis is a alpheid shrimp with a large snapping claw. When this claw is forcibly closed, it results in a loud snap sound. Since underwater sound propagates fast over long distance, it has been hypothesized that the snapping sound may be used for underwater acoustic communication. This work is designed to test the “acous- tic communication” hypothesis. Individual shrimp were housed in a rectangular tank and a plastic rod was used to prod to initiate snapping action. A hydrophone was used to record the snapping sound. The recorded sounds were analyzed with a computer-based SIGNAL analysis system. Time, frequency signal, and spectrogram corre- Abstracts, 1996 Annual Meeting 53 lations were compared between sounds of different indi- viduals (males, females). Playback of the snapping sound was also performed on 10 shrimp to initiate snapping ac- tion. Snapping activity of two encountering individuals un- der various conditions (light, dark, amputation of anten- nular and antennal flagellum) was also videotaped and an- alyzed. Sound similarity analyses showed no significant difference in acoustic signatures made by different ani- mals. Playback of the snapping sound failed to initiate any snapping action. Contacts of antennular or antennal fla- gellum of two encountering shrimp before snapping oc- curred were observed in 256 snappings (out of a total of 268). For amputated shrimp physical contact of body parts was required before snapping actions were initiated. The results show that the snapping sound is not used for acous- tic communication function. It is likely that the strong underwater jet currents created by the snapping action are used by the shrimp to ward off intruders. The snapping sound produced is the by-product of the snapping action. Supported by Howard Hughes Medical Institute and a University of Kentucky Undergraduate Research Creative Grant. Comparison of a hybrid (Heliconia latispatha X imbri- cata; Heliconiaceae) with its parent species with respect to extrafloral nectaries. C. TONY R. HAMPTON* and THOMAS C. RAMBO, Department of Biological Sci- ences, Northern Kentucky University, Highland Heights, KY 41099. As part of on-going studies of the role of extrafloral nectaries on buds of Heliconia latispatha and of the in- sects visiting these nectaries, we studied a hybrid of H. latispatha (which has nectaries) and H. imbricata (which lacks nectaries). We studied a set of 10 H. latispatha, 10 H. imbricata, and 8 hybrids, all growing along the edge of the landing strip at Estacién Sirena in Parque Nacional de Corcovado, Costa Rica. We recorded the insect visitors to the buds hourly from 0530 to 0930 and from 1330 to 1730, the time of most insect visitation, for 5 days. Heli- conia latispatha had significantly more insects (primarily ants) on the buds; H. imbricata and the hybrid had few to no insects on the buds. The buds of H. latispatha and the hybrid are similar in color, so color is not primarily responsible for the attraction of insects. We hypothesize that the insect attraction by H. latispatha is chemical, and that the genes for this attraction must be recessive. Comparison of two species of Heliconia (Heliconiaceae) with respect to the presence of extrafloral nectaries. THO- MAS C. RAMBO and C. TONY R. HAMPTON, De- partment of Biological Sciences, Northern Kentucky Uni- versity, Highland Heights, KY 41099. In previous studies we have shown that the platanillo, Heliconia latispatha, produces nectar on the outside of bracts of terminal buds in an inflorescence. The nectar, secreted onto the bud’s surface, is utilized by a variety of insects, including ants, mosquitoes and other dipterans (Dolichopodidae, Richardidae, Neriidae), crickets, roach- es, lygaeid bugs, and wasps. To clarify the function of these extrafloral nectaries we compared the insect visitors to Heliconia latispatha with visitors to the buds of H. im- bricata, a species growing close to H. latispatha. We re- corded the visitors to 15 buds of each species hourly from 0530 to 0930 and from 1330 to 1730, the time of most insect visitation, for 5 days. Heliconia imbricata attracted significantly fewer insects than H. latispatha. In fact H. imbricata attracted so few insects that we conclude that it does not produce nectar on the buds. We will discuss possible reasons for this difference and their implications for determining the function of the extrafloral nectaries on H. latispatha. Effects of cannibalism on population structure of the marine cave isopod Bahalana geracei from Lighthouse Cave, San Salvador Island, Bahamas. JERRY H. CAR- PENTER and RONALD D. BITNER,* Department of Biological Sciences, Northern Kentucky University, High- land Heights, KY 41099. Cannibalism by Bahalana geracei was observed once in Lighthouse Cave, in July 1995: a 11.0 mm long female was eating a 6.5 mm long female. We performed experi- ments to investigate factors influencing cannibalism in this troglobitic species. From August to November 1996, two to four specimens were placed together in small plastic bowls, maintained without food in incubators at cave tem- perature (26°C), and observed daily. We never witnessed cannibalism while it was occurring in the lab; we did find remains of six cannibalized victims (some still alive)— translucent exoskeletons of cannibals revealed remains of victims inside. The following cannibal/victim combinations show the variety of sizes: (1) 3.2 mm second-instar manca (baby)/2.7 mm first-instar manca (sex of mancas could not be determined), (2) 4.2 mm fourth-instar juvenile female/ 2.7 mm first-instar manca, (3) 8.2 mm female/8.2 mm fe- male slightly damaged in another predator-prey experi- ment, (4) 14.3 mm female and 7.4 mm female/7.5 mm, (5) 14.3 mm female/7.4 mm female (same individuals as above), and (6) 15.7 mm female and 13.9 mm female/12.4 mm female. These cannibalisms occurred after specimens were together several days, except case #3, which oc- curred within 1 day. In at least 10 additional cases, two to four specimens of various sizes and sexes were together >30 days without cannibalism. We conclude that canni- balism in B. geracei is unpredictable. It is more likely when size difference is large. Cannibalism in B. geracei seems less common than in many surface-dwelling arthro- pods (e.g., spiders and other marine isopods). Cannibalism is reduced by B. geracei’s starvation resistance and slow metabolism. Nevertheless, cannibalism is probably fre- quent enough in this long-lived species to significantly af- fect the population structure. Female preferences for stimulatory male odors in the prairie vole (Microtus ochrogaster). JILL H. KRUPER* and TERRY L. DERTING, Department of Biological Sci- ences, Murray State University, Murray, KY 42071. 54 Transactions of the Kentucky Academy of Science 58(1) Physical contact with conspecific pheromones or urine of males is necessary for reproductive stimulation of fe- male prairie voles. We investigated whether female prairie voles (Microtus ochrogaster) of different ages and repro- ductive status prefer odors from males who are most likely to stimulate female reproductive activity. We compared odors from intact and castrated adult males because urine from intact males is known to be a more effective stim- ulant of female reproductive activity than is urine from castrated males. Preferences of females for male odors were determined using a Y-maze olfactometer or nesting apparatus. At 7-10 d post-surgery, young anestrus females significantly preferred the odors of intact males as com- pared to the odors of castrated males. At 2 months post- surgery, no significant preference for odors of intact or castrated males was exhibited by older anestrus females. Older females in estrus, however, tended to prefer the odor of intact males. Young and old anestrus females showed no significant preference for the odors of castrat- ed or intact males during nesting-preference tests; how- ever, young females in estrus tended to discriminate against the odors of castrated males. Our results indicated that young females, and those in estrus, were most likely to discriminate among male odors, exhibiting less prefer- ence for odors from castrated males than intact males. Male odors indicative of reproductive potential may, therefore, play a role in female mate-choice in prairie voles depending upon the age and reproductive status of females. Impacts of dam construction on densities and distri- butions of bald eagles, Haliaeetus leucocephalus, Accipit- ridae) along the Ohio River shoreline between Paducah, Kentucky, and Cairo, Illinois. VINCENT EVIN STAN- FORD* and TERRY L. DERTING, Department of Bi- ology, Murray State University, Murray, KY 42071. Shoreline densities and distributions of wintering bald eagles (Haliaeetus leucocephalus) were compared to ex- amine the impacts of an ongoing lock-and-dam construc- tion project on an Ohio River segment 65 km long be- tween Paducah, KY, and Cairo, IL. Low-altitude flights were conducted once weekly for 10 weeks from mid-De- cember through mid-March in winters of 1987/1988 (pre- dam construction) and 1994/1995 (ongoing dam construc- tion). Densities and distributions of bald eagles per river quarter-mile were recorded during the survey flights. Pro- portional densities of bald eagles were compared between the two survey periods in a 1- and a 6-mile interval with the site of dam construction centered in each interval. Proportional densities of bald eagles in the 1-mile interval, during the 1994/1995 surveys, were significantly reduced from proportional densities that occurred during the 1987/ 1988 surveys (P = 0.0024). A marginally significant re- duction in proportional densities of bald eagles occurred in the 6-mile interval during the 1994/1995 surveys when compared with the 1987/1988 surveys (P = 0.07). A mar- ginally significant shift in %-mile distributions along the entire segment also occurred between the two survey pe- riods (P = 0.06). These results indicated that bald eagles were avoiding the area of dam construction and that over- all bald eagle distributions may be shifting away from the area of dam construction. We recommend that further studies be conducted to examine future impacts on bald- eagle densities and distributions as construction proceeds and to determine long-term impacts of dam placement on bald eagles. Nest-site selection and leaf-nest composition of Sciurus carolinensis (Sciuridae) in continuous and isolated woods in western Kentucky. JAMES S. ARMSTRONG* and TERRY L. DERTING, Department of Biological Sci- ences, Murray State University, Murray KY 42071. Location and composition of grey squirrel (Sciurus car- olinensis) leaf-nests were analyzed in three different forest habitats: old (116-125 years) continuous-forest (3 sites, 2 ha each); young (85-92 years) continuous-forest (3 sites, 2 ha each); and isolated woodlot (106 years; 1 site, 3 ha). The isolated woodlot contained a significantly higher num- ber of small (DBH 3-25 cm) and total number of trees than either continuous-forest habitat. No significant dif- ferences in vegetational cover existed among habitats. The isolated woodlot had significantly higher nest density (12.5 nests/ha) than the old or young continuous-forest habitats (3.2 + 0.9 nests/ha and 3.3 + 0.3 nests/ha, respectively). The density of grey squirrels in the isolated woodlot (13.2 squirrels/ha) was also significantly greater than squirrel densities in old and young continuous-forest habitats (1.9 + 0.6 and 1.8 + 1.0 squirrels/ha, respectively). A signifi- cant positive correlation existed between leaf-nests/ha and grey squirrels/ha across the seven study sites. Grey squir- rels chose significantly fewer small size trees (DBH <20 cm) and significantly more medium size trees (DBH 20- 50 cm) for leaf-nest placement than expected based on tree availability. Grey squirrels chose large size trees (DBH > 50 cm) for leaf-nest placement in accordance with tree availability. Tree species selected and the pre- ferred DBH size for nest location were positively corre- lated in the young continuous-forest and isolated woodlot habitats. The composition of leaf-nests did not differ among habitats although the leaf-nests in the old contin- uous-forests had the greatest dry masses. Guidelines for Contributors to the Transactions 1. GENERAL . Original papers based on research/review in science will be considered for publication in the Transactions; at least the first author must be a member of the Acad- emy. Announcements, news, and notes will be includ- ed as received. . Papers (in triplicate) may be submitted at any time to the editor. John W. Thieret Biological Sciences Northern Kentucky University Highland Heights, KY 41099 Phone: (606) 572-6390 FAX: (606) 572-5639 E-mail: thieretj}@nku.edu List in the cover letter your telephone/FAX numbers, your E-mail address, and the names, addresses, and phone numbers of two persons who are potential re- viewers. , Format/style of papers must conform to practices in recent issues of the Transactions, which are, in effect, a style manual. The running head at top right of each page should give name of author(s), a short version of paper title, and page number of total. Do not staple pages together. . Papers should be submitted in hard copy and on a 3.5 inch disk, preferably in WordPerfect for Windows 6.1 or earlier version. . Indent the first line of each paragraph (but not the first line of entries in the Literature Cited). 2. FORMAT Papers should be in 10 cpi type on white paper 8.5 x 11 inches, with margins at least 1 inch all around. Double-space throughout the paper (i.e., one full line of space between each two lines of text, literature cit- ed, or tabular data). Do not justify right margins. . Except for scientific names of genera and of infra-ge- neric taxa, which should be typed in italics, the same type (roman) should be used throughout (i.e., one type size only, no bold). . Sequence of sections in papers should, where appro- priate, be as follows: title of paper, name/address of author(s), abstract, body of paper, footnotes, table cap- tions, figure captions (all the preceding on consecu- tively numbered pages), tables, and figures. . The first page should include the running head and, centered near the top of the sheet, the paper's title and the name and address of author(s). These should be followed immediately by the abstract. . The abstract, not to exceed 200 words, should be con- cise, descriptive, and complete in itself without refer- ence to the paper. . The body of the paper should, where appropriate, in- clude the following sections: Introduction, Materials and Methods, Results, Discussion, Summary, Acknowl- edgements, and Literature Cited. . No more than three levels of headings should be used: level 1, in capitals, centered; level 2, in capitals/low- ercase, flush left; level 3, in italics, a paragraph indent, with initial capital only (except proper nouns and ad- jectives), and followed by a period, the text then start- ing after one blank space. . Personal communications (avoid if possible) should be indicated in the text as follows: (name, affiliation, pers. comm., date), e.g., (O.T. Mark, Wainwright College, pers. comm., 5 Jun 1995). Notes should follow the format established in Trans- actions 57(1). 3. STYLE . In text, spell out one-digit numbers unless they are used with units of measure (four oranges, 4 cm) and use numerals for larger numbers; do not begin any sentence with a numeral. . Use no footnotes except those for title page and tables. Footnotes, identified by consecutive superscript num- bers, should be entered on a separate sheet. . Measurements should be in metric and Celsius units. Define lesser-known symbols and give the meaning of acronyms at first use. Express time of day in the 24- hour system. Dates should be written day, month (ab- breviated to three letters), year without internal punc- tuation. Units with multiple components should have individual components separated by a virgule (e.g., g/ m? or g/m?/yr). . Names of authors of binomials may be included but only at the first mention of the binomial. Cultivar names are not italicized but are enclosed in single quotes or preceded by cv. . Useful guides for contributors to the Transactions are the following: Scientific style and format: the CBE manual for authors, editors, and publishers, 6th ed., Cambridge University Press, 1994; The Chicago man- ual of style, 14th ed., University of Chicago Press, 1993; The ACS style guide, American Chemical Soci- ety, Washington, DC, 1986; and AIP style manual, American Institute of Physics, New York, 1990. 4, IN-TEXT CITATION OF LITERATURE . Cite publications in the text by author(s) and date— e.g., (Readley 1994); multiple citations should be in alphabetical order and separated by semi-colons—e.g., (Ashley 1987; Brown 1994; Foster 1975); multiple ci- tations of works by one author(s) should be in chro- nological order—e.g., (Jones 1978, 1983); publications by one author(s) in the same year should be distin- guished by a, b, c, ete—e.g., (Smith 1994a, 1994b). For in-text references to works with one to three au- thors use names of all authors—e.g., (Jones, Smith, and Williams 1991); for works with four or more au- Transactions of the Kentucky Academy of Science 58(1) thors use name of the first author followed by et al.— e.g., (Lee et al. 1985). B. Do not include any reference unless it has been pub- lished or accepted for publication (“in press”). In the latter case give the name of the accepting journal or the publisher/place of publication; use n.d. in place of a date for in-text citation of “in press” references, e.g., (Jones n.d.). 5. LITERATURE CITED A. List all authors of each entry. Do not abbreviate jour- nal titles; abbreviations for these will be supplied by the editor. B. The first line of each reference should be typed flush left; the remaining lines should be indented. C. Examples of common types of references are given below. JOURNAL ARTICLE Lacki, M.J. 1994. Metal concentrations in guano from a gray bat summer roost. Transactions of the Kentucky Academy of Science 55:124-126. BOOK Ware, M., and R.W. Tare. 1991. Plains life and love. Pi- oneer Press, Crete, WY. PART OF A BOOK Kohn, J.R. 1993. Pinaceae. Pages 32-50 in J.F. Nadel (ed). Flora of the Black Mountains. University of Northwestern South Dakota Press, Utopia, SD. WORK IN PRESS Groves, S.J., LV. Woodland, and G.H. Tobosa. n.d. De- serts of Trans-Pecos Texas. 2nd ed. Ocotillo Press, Yucca City, TX. 6. ILLUSTRATIONS FIGURES (LINE DRAWINGS, MAPS, GRAPHS, PHO- TOGRAPHS) Figures must be camera-ready, glossy, black-and-white prints of high quality or laser prints of presentation qual- ity. These should be designed to use available space ef- fectively: a full page or part of one, or a full column or part of one. They should be mounted on heavy white board and covered with a protective sheet of paper; pho- tographs to be grouped as a plate should have no space between them. Dimensions of plates must observe page proportions of the journal. Each illustration in a plate may be numbered as a separate figure or the entire plate may be treated as one figure. Include scale bars where appro- priate. Lettering should be large enough to be legible after reduction; use lowercase letters for sections of a figure. Figure captions should be self-explanatory without refer- ence to the text and should be entered on a page separate from the text. Number figures in Arabic numerals. Statis- tics presented in figures should be explained in the caption (e.g., means are presented ce Siam a) TABLES Each table and its caption must be double-spaced, num- bered in Arabic numerals, and set on a sheet separate from the text. The caption should begin with a title relat- ing the table to the paper of which it is a part; it should be informative of the table’s contents. Statistics presented in the table should be explained in the caption (e.g., means are presented + SE, n = 7). Table should be sub- mitted in hard copy only; they need not be included on the disk. 7. PROOFS Authors are responsible for correcting proofs. Extensive alterations on proofs are expensive; costs will be assessed to authors. Proofs must be returned to the editor within 3 days after the author receives them; delay in return may result in delay of publication. 8. REPRINTS Forms for ordering reprints will be sent to the author when the proofs are sent. They are to be returned directly to Allen Press, not to the editor. 9. ABSTRACTS FOR ANNUAL MEETINGS Instructions on style of abstract preparation for papers presented at annual meetings may be obtained from the editor. Copies will be available also at each annual meet- ing of the Academy. NEWS KAS Annual Meetings The 1997 annual meeting of the Kentucky Academy of Science will be held 13-15 November at Morehead State University, Morehead; the me meeting, at Jefferson Community College, Southwest Campus, Louisville. PUBLICATION Wildflowers of Mammoth Cave National Park, by Randy Seymour, has just been published. A soft-cover book of 254 pages, it illustrates by excellent color photographs about 400 wild- flowers, each with accompanying text. Appendices show flowering times, where in the park the various flowers have been seen, and the number of species noted by Seymour along each of the park’s trails. Order from bookstores or directly from The University Press of Kentucky, 663 South Limestone Street, Lexington, KY 40508-4008; ISBN 0-8131-0898-5; price $17.95 (+$3.50 shipping). CONTENTS Relationships Among Habitat, Cover, and Eastern Cottontails (Sylvila- gus floridanus) in Kentucky. William M. Giuliano, Charles L. Elliott, and: Jeffery: D: Sole) oe Se ee, ONG SS OEN Oia Nae tae Ne tn atic aa Separation of Spawning Habitat in the Sympatric Snubnose Darters Eth- eostoma flavum and E. simoterum (Teleostei, Percidae). Jean C. Por- Fer field esc cs ee a ee Sa oo ak Wade eee cata Soya R Us PAPA ay ae Common Names of Vascular Plants Reported by C.S. Rafinesque in an 1819 Descriptive Outline of Four Vegetation Regions of Kentucky. Ron- ald L. Stuckey and James S. Pringle .............0ccccccececcnceccecssecceccevece Some Comments on Constantine Rafinesque’s 1819 Description of Bo- tanical Regions in Kentucky. William S. Bryant ................0ccccseeceeeees Effect of Light on Daily Emergence of Cercariae of the Trematodes Echi- nostoma trivolvis (Echinostomatidae) and Cephalogonimus vesicau- dus (Cephalogonimidae) from Natural Infections of the Snail Helisoma trivolvis (Planorbidae) at Owsley Fork Reservoir, Kentucky. Ron Rosen, Peter Blair, Jeff Ellington, and Jason Backus .................0cceccseeseees Classifying Free Bieberbach Groups. Raymond F. Tennant ............... DISTINGUISHED SCIENTIST AND OUTSTANDING TEACHER AWARDS OG ee RSA cas caueceea ea cnn een ceee ns dere COU se oda ral oun ne ean ne Meg NOTES Human Myiasis in Kentucky Caused by Cuterebra sp. (Diptera: Oestri- dae): Lee Hs Townsend oo ee ee a ee Wea a Ree ae ABSTRACTS OF SOME PAPERS PRESENTED AT THE 1996 ANNUAL MEET- ING OF THE KENTUCKY ACADEMY OF SCIENCE ..................cecesceceeceees TRANSACTIONS. vx | THE wee NIT LICK Y ACADEMY OF toi NCE i sg Volume 58 Number 2 September 1997 Official Publication of the Academy — The Kentucky Academy of Science Founded 8 May 1914 GoverninGc Boarp For 1997 Executive COMMITTEE President: Marcus T. McEllistrem, Department of Physics and Astronomy, University of Kentucky, Lexing- ton, KY 40506-0055 President Elect: Patricia K. Doolin, Research and Development, Ashland Petroleum Company, P.O. Box 391, Ashland, KY 41114 Vice President: Gordon K. Weddle, Department of Biology, Campbellsville University, Campbellaville: KY 42718 Past President: William S. Bryant, Department of Biology, Thomas More College, Crestview Hills, KY 41017 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 Treasurer Elect: William E. Houston, 161 Morningstar Court, Bowling Green, KY 42103 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; (606) 572-6390 Editor, NEWSLETTER (ex officio): Maria K. Falbo-Kenkel, Department of Physics and Geology, Northern Kentucky University, Highland Heights, KY 41099 Chair, Junior Academy of Science (ex officio): Vincent A. DiNoto Jr., Department of Physics, Jefferson Community College SW, 1000 Community College Drive, Louisville, KY 40272 Program Coordinator (ex officio): Robert O. Creek, Department of Biological Sciences, Eastern Kentucky University, Richmond, KY 40475 MEMBERS, GOVERNING BOARD Robert J. Barney 1999 Barbara L. Rafaill 1999 Gerald L. DeMoss 1997 J. G. Rodriguez 1998 James F. Hopgood 1998 AAAS/NAAS Representative Bruce A. Mattingly 2000 Lee A. Roecker 2000 Wimberly C. Royster 1997 COMMITTEE ON PUBLICATIONS Editor and John W. Thieret, Department of Biological Sciences, Northern Kentucky University, Chair: Highland Heights, KY 41099 Associate Editor: James O. Luken, Department of Biological Sciences, Northern Kentucky University, Highland Heights, KY 41099 Index Editor: Varley Wiedeman, OPE of Biology, University of Louisville, Louisville, KY 40292 Editorial Board: William S. Bryant, Bence of Biology, Thomas More College, Crestview Hills, KY 41017 John P. Harley, Department of Biological Sciences, Eastern Kentucky Uhre Richmond, KY 40475 Marcus T. McEllistrem, Department of Physics and Astronomy, University of Kentucky, Lexington, KY 40506-0055 All manuscripts and correspondence concerning manuscripts should be addressed to the Editor. The TRANSACTIONS are indexed in BIOSIS and in State Academies of Science Abstracts. 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: $45.00 domestic; $50.00 foreign. Back issues are $30.00 per volume. The TRANSACTIONS are issued semiannually in March and September. Two numbers comprise a volume. Correspondence concerning memberships or subscriptions should be addressed to the Secretary. This paper meets the requirements of ANSI/NISO Z39.48-1992 (Permanence of Paper). 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 Bellarmine College Hazard Community College Berea College Paducah Community College Campbellsville College Somerset Community College Centre College Southeast Community College Cumberland College Associate Member Ashland Community College Maysville Community College Georgetown College Midway College Jefferson Community College | Owensboro Community College Kentucky State University Spalding University Kentucky Wesleyan College Thomas More College Transylvania University INDUSTRIAL AFFILIATES Associate Patron Ashland Oil, Inc. Member Corhart Refractories Corporation Hoechst Celanese Corporation MPD, Inc. United Catalyst, Inc. Westvaco Associate Member All-Rite Pest Control Lockheed Martin Wood Hudson Cancer Research Laboratory, Inc. 7 tne TRANSACTIONS of the KENTUCKY ACADEMY of SCIENCE September 1997 Volume 58 Number 2 Trans. Ky. Acad. Sci. 58(2):55-59. 1997. Continuing Decline in the Freshwater Unionid (Bivalvia: Unionidae) Fauna in the Cumberland River Downstream from Cumberland Falls, Kentucky Ronald R. Cicerello and Ellis L. Laudermilk Kentucky State Nature Preserves Commission, 801 Schenkel Lane, Frankfort, Kentucky 40601 ABSTRACT The freshwater unionids in the Cumberland River downstream from Cumberland Falls in southeastern Kentucky were sampled to determine the status of the fauna relative to previous studies. A comparison of collections by Wilson and Clark in 1910-1911, Neel and Allen in 1947-1949, and Stansbery in 1961 revealed the loss of 6 of the 22 species known from the site and reduced abundance in several others. We found only 10 live species, all less abundant than during previous studies, and no shells. The continuing decline in the fauna is attributable to habitat degradation and loss associated with coal, mining, general watershed devel- opment, and impoundment of the river to create Lake Cumberland. INTRODUCTION Long-term monitoring can provide status and trend information needed for conserva- tion of freshwater unionids. To determine changes in the Cumberland River fauna down- stream from Cumberland Falls in southeast- ern Kentucky following the creation of Lake Cumberland, Stansbery (1969) sampled the unionids and compared his findings to those of Wilson and Clark (1914) and Neel and Al- len (1964). Wilson and Clark (1914) found 20 species, 10 of which were common or abun- dant, at this site in 1910-1911 while evaluating the Cumberland River as a source of unionids to replace upper Mississippi River populations overharvested for button manufacturing and pearls. During 1947-1949, just prior to the closure of Wolf Creek Dam and the formation of Lake Cumberland, Neel and Allen (1964) encountered 15 species at the site, 13 of which were common or abundant. Stansbery (1969) collected below the falls in 1961 and noted that changes in faunal com- position prior to Neel and Allen (1964) were 50 attributable to almost 40 years of increased ac- ids from coal mining and washing above the falls. Changes after 1949 may have resulted from continuing impacts from coal mining op- erations and the influence of Lake Cumber- land. By 1947-1949, Cyclonaias tuberculata, Ellipsaria lineolata, Obliquaria reflexa, and Ptychobranchus subtentum were lost from the site, but Villosa iris had been found. Between 1949 and 1961, Lampsilis cardium and Trun- cilla truncata were lost, 6 of the 16 taxa found by Stansbery (1969) had decreased in abun- dance, and the remainder were stable or had increased in number. We resampled this site to determine the present status of the fauna. STUDY AREA Cumberland Falls, a 17-m-high barrier to the upstream movement of aquatic organisms (Burr and Warren 1986; McGrain 1966), is lo- cated at river km 904.9 in McCreary and Whitley counties in southeastern Kentucky (Figure 1). Land use in the 5120 km? water- shed (Bower and Jackson 1981) is ca. 84% for- 56 Transactions of the Kentucky Academy of Science 58(2) River km 898.6 Dog Slaughter Creek —> Z McCreary County Whitley County Figure 1. Cumberland River study area downstream from Cumberland Falls, McCreary and Whitley counties, Eagle Creek Cumberland Falls ~ River km 904.9 Kentucky. Inset shows study area location in the state. Shaded areas were searched for unionids. est, 13% agriculture, 2.5% mining, and 0.5% urban and developed areas (Mayes, Sudderth, and Etheredge 1975). Downstream from the falls, the river flows through a forested and boulder-lined gorge and is impounded by the backwaters of Lake Cumberland at approxi- mate river km 898.6, depending on reservoir pool elevation. The Cumberland River was impounded at river km 742 in 1950 to create Lake Cumberland. At low flow, river width ranges from ca. 5 to 24 m in riffles and pools, respectively, and depth exceeds 5 m in pools and 1 m in riffles. Pools and riffles are com- posed of large boulders, cobbles, and exten- sive deposits of coal fines and sand intermixed with lesser quantities of gravel, pebbles, coal, and trash in eddies and areas sheltered from swift current. During low flow, huge deposits of sand and coal fines are exposed in the plunge basin below the falls. Mean discharge on the sampling dates was 7.4 m*/sec, consid- erably less than the mean of 26.6 m*/sec for July-September, when streamflow is at or near the annual low, and the mean annual discharge of 89.7 m*/sec (USGS 1993). Water quality is improving, but the upper Cumberland River continues to be impacted by pollutants asso- ciated with coal mining (e.g., acid mine drain- age, silt, and metals), domestic waste, highway construction, and poor land use (Harker et al. 1980; Kentucky Division of Water 1994a, 1994b). MATERIALS AND METHODS In 1987, 1989, and twice in 1993, we ex- amined a 0.5-km-long river segment from the base of Cumberland Falls to Eagle Creek for unionids (Figure 1). We made additional col- lections 0.5 km downstream from Eagle Creek and near the mouth of Dog Slaughter Creek to further gauge the status of the fauna. Dur- ing each visit, one or two persons searched 2 or more hours for shells and live specimens by snorkeling and with waterscopes, and the shoreline was examined for shells. Nomencla- ture follows Hoeh (1990) and Turgeon et al. (1988). RESULTS AND DISCUSSION We found a total of 10 species, a loss of six taxa when compared to Stansbery’s (1969) re- sults and an overall 54% reduction in the fau- na compared to earlier studies (Table 1). With the exception of Elliptio crassidens, all taxa were less abundant than in 1961. Among the taxa lost are Villosa trabalis, a USFWS (1994) endangered species, and Lampsilis ovata, Pty- chobranchus subtentum, and Villosa lienosa, which are rare in Kentucky (KSNPC 1996). Others that have declined or have been lost (e.g., Actinonaias ligamentina, Elliptio dilata- ta, Lampsilis cardium, Potamilus alatus, Quadrula pustulosa, Tritogonia verrucosa) have broad habitat requirements (Gordon and Layzer 1989) and often are dominant mem- bers of unionid communities (Houslet 1996; Smathers 1990). Corbicula fluminea, an intro- duced exotic bivalve, was uncommon. Observations by Wilson and Clark (1914) and Neel and Allen (1964) provide anecdotal evidence of the decline in unionid abundance at the falls. Wilson and Clark (1914) reported that unionids were “usually found crowded about the base of large rocks” where they were “easily accessible to their enemies, es- pecially during low water, and many of them are killed by muskrats, raccoons, mink, and oc- casional otter.” Neel and Allen (1964) found Cumberland Unionids—Cicerello and Laudermilk aa Table 1. in four studies: 1910-1911, 1947-1949, 1961, 1987-1993. 1910-1911! Species Actinonaias ligamentina (Lamarck) 194 Actinonaias pectorosa (Conrad) We Cyclonaias tuberculata (Rafinesque) 2 Ellipsaria lineolata (Rafinesque) P Elliptio crassidens (Lamarck) 57 Elliptio dilatata (Rafinesque) 122 Lampsilis cardium (Rafinesque) 0 Lampsilis fasciola Rafinesque 16 Lampsilis ovata (Say)? 49 Lasmigona costata (Rafinesque) 4] Ligumia recta (Lamarck) 8 Medionidus conradicus (Lea) P Obliquaria reflexa Rafinesque 8 Potamilus alatus (Say) 8 Ptychobranchus fasciolaris (Rafinesque) 81 Ptychobranchus subtentum (Say)° 12 Quadrula pustulosa (Lea) 49 Tritogonia verrucosa (Rafinesque) 32 Truncilla truncata Rafinesque 16 Villosa iris (Lea) 0 Villosa lienosa (Conrad)* P Villosa trabalis (Conrad)* 4] Total species 20 Total individuals 810 Freshwater unionids collected from the Cumberland River downstream from Cumberland Falls, Kentucky, 1947-19497 1961 1987-1993" Number collected A 39 2 A 161 >50 0 0 0 0 0 0 C (2) 2 A 113 i A 0 0 C 20 0 0 10 0 0 1(?) 2) A it 1 A 154 0 0 0 0 E 28 5 A 35 5 0 0 0 A 122 10 A 75 4 R 0 0 R Oi 0 0) 9 0 A vi 0 15 16 10 a= 810 ca. 88 ' Wilson and Clark (1914); * Neel and Allen (1964); > Stansbery (1969); * maximum number observed alive on any sampling date during the present study; > USFWS (1994) and/or KSNPC (1996) listed species. A = abundant; C = common; R = rare; P = present; 0 = none taken; (2) = subfossil remains only, “great numbers of mussels, many of which were quite large and very old” in sand pockets among large stones and slabs. We found a con- centration of live unionids only in a small area near the mouth of Eagle Creek. No shells de- posited by floodwater or left by predators were found along the shoreline. In the river 0.5 km downstream from Eagle Creek and near the mouth of Dog Slaughter Creek, we found limited numbers (in order of abundance) of only Actinonaias pectorosa, Tri- togonia verrucosa, Actinonaias ligamentina, Elliptio dilatata, Potamilus alatus, Ptycho- branchus fasciolaris, and Quadrula pustulosa. These sites are within the river segment ex- tending from “Anvil Shoals,” 1.6 km below the falls, to Burnside, Kentucky, that mussel fish- erman reported was full of mussels (Wilson and Clark 1914). The fauna of this last remaining free-flow- ing segment of the Cumberland River below the falls in southeastern Kentucky is now com- prised largely of species with broad environ- mental tolerances (Dennis 1984; Gordon and Layzer 1989). The change in the fauna is at- tributable to pollutants associated with coal mining and to the impoundment of the river (Stansbery 1969), and general watershed de- velopment (e.g., road building, channelization, urbanization). Recolonization is precluded by Lake Cumberland, which isolates remnant unionid populations in Buck Creek (Schuster, Butler, and Stansbery 1989), the Rockcastle River (Cicerello unpubl. data; Thompson 1985), and the Big South Fork Cumberland River (Bakaletz 1991), and which acts as a bar- rier to the movement of host fishes. Direct tributaries to the area (e.g., Eagle and Dog Slaughter creeks) do not support unionid pop- ulations, and the Cumberland River above Cumberland Falls supports a depauperate fau- na comprised of 11 species that generally are uncommon in the basin (Cicerello unpubl. data; Cicerello, Warren, and Schuster 1991). The unionid community in the river below Cumberland Falls is the last and best remain- ing vestige of the diverse fauna that histori- cally inhabited the mainstem Cumberland River in southeastern Kentucky. Downstream from Wolf Creek Dam, fluctuating levels of 58 Transactions of the Kentucky Academy of Science 58(2) cold, turbulent water released from Lake Cumberland preclude unionid reproduction, and only Cumberlandia monodonta Say and Cyclonaias tuberculata persist (Cicerello per- sonal observation; Miller, Rhodes, and Tippit 1984). Lake Cumberland backwaters in the lower Big South Fork Cumberland and Rock- castle rivers, and the mainstem are heavily silt- ed and have yielded Leptodea fragilis Rafin- esque, Potamilus ohiensis Rafinesque, Pygan- odon grandis Say, and Utterbackia imbecillis Say. Only Potamilus alatus and P. ohiensis in- habit the lower embayed segment of Buck Creek (Schuster, Butler, and Stansbery 1989). Perhaps 20 (31%) of the 65 taxa that histori- cally inhabited the mainstem Cumberland River remain (Cicerello unpubl. data). Monitoring in the river downstream from Cumberland Falls in the future will provide additional information about the impact of Lake Cumberland and watershed land use on the unionid fauna. However, the survival of Kentucky's remnant Cumberland River union- id community, which includes members of the unique Cumberlandian fauna (Ortmann 1924) (e.g., Epioblasma brevidens, E. capsaeformis, Ptychobranchus subtentum, and Villosa tra- balis) found nowhere else in the state, is de- pendent upon the protection of Buck Creek, the Rockcastle River, and the Big South Fork Cumberland River. ACKNOWLEDGMENTS We acknowledge the field assistance of Ste- phen McMurray (Eastern Kentucky Universi- ty). G.A. Schuster (Eastern Kentucky Univer- sity) critically reviewed the manuscript and provided valuable suggestions. This study was supported in part by the Kentucky Depart- ment for Surface Mining Reclamation and En- forcement, Frankfort, Kentucky. LITERATURE CITED Bakaletz, S. 1991. Mussel survey of the Big South Fork National River Recreation Area. Master's thesis, Ten- nessee Technological University, Cookeville, TN. Bower, D.E., and W.H. Jackson. 1981. Drainage areas of streams at selected locations in Kentucky. Open File Report 81-61. Geological Survey, U. S. Department of the Interior. Burr, B.M., and M.L. Warren Jr. 1986. A distributional atlas of Kentucky fishes. Kentucky State Nature Pre- serves Comm. Sci. Tech. Ser. 4. 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: 113-129. Dennis, $.D. 1984. Distributional analysis of the fresh- water mussel fauna of the Tennessee River system, with special reference to possible limiting effects of siltation. Ph. D. dissertation, Virginia Polytechnic Institute and State University, Blacksburg, VA. Gordon, M.E., and J.B. Layzer. 1989. Mussels (Bivalvia: Unionoidea) of the Cumberland River: review of life histories and ecological relationships. U. S. Fish and Wildlife Service Biological Report 89(15). 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 Comm., Tech. Rept., Frankfort, KY. Hoeh, W.R. 1990. Phylogenetic relationships among east- ern North American Anodonta (Bivalvia: Unionidae). Malacol. Rev. 23:63-82. Houslet, B.S. 1996. Age, growth, and stability of a mussel assemblage in Horse Lick Creek, Kentucky. Master's thesis, Tennessee Technological University, Cookeville, TN. Kentucky Division of Water. 1994a. Cumberland River biological and water quality investigation. Wild River segment. Kentucky Division of Water, Frankfort, KY. Kentucky Division of Water. 1994b. 1994 Kentucky re- port to congress on water quality. Kentucky Division of Water, Frankfort, KY. [KSNPC] Kentucky State Nature Preserves Commission. 1996. Rare and extirpated plants and animals of Ken- tucky. Trans. Kentucky Acad. Sci. 57:69-91. Mayes, Sudderth, and Etheredge, Incorporated. 1975. The river basin water quality management plan for Kentucky. Upper Cumberland River. Prepared for Ken- tucky Division of Water, Frankfort, KY. McGrain, P. 1966. Geology of the Cumberland Falls State Park area. Kentucky Geological Survey Series X, Spec. Publ. IL. University of Kentucky, Lexington, KY. 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 W.R. Allen. 1964. The mussel fauna of the upper Cumberland basin before its impoundment. Malacologia 1:427-459. Ortmann, A.E. 1924. The naiad-fauna of the Duck River in Tennessee. Am. Midl. Naturalist 9:1S-62. 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. Kentucky Acad. Sci. 50:79-85. Smathers, K.L. 1990. An analysis of a bivalve (Mollusca: Bivalvia) community in the Licking River, at Moores Ferry, Kentucky. Master’s thesis, Eastern Kentucky University, Richmond, KY. Stansbery, D.H. 1969. Changes in the naiad fauna of the Cumberland Unionids—Cicerello and Laudermilk 59 Cumberland River at Cumberland Falls in eastern Ken- tucky. Bull. Am. Malacol. Union 1969:16-17. Thompson, Y.L. 1985. The mussel fauna of the Rockcastle River system, Kentucky (Bivalvia: Unionidae). Master's thesis, Eastern Kentucky University, Richmond, KY. 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. 16:1—277. [USFWS] United States Fish and Wildlife Service. 1994. Endangered and threatened wildlife and plants. De- partment of the Interior, Washington, DC. [USGS] United States Geological Survey. 1993. Water re- sources data—Kentucky. Water year 1993. Data Report KY-93-1, Louisville, KY. 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 Document 781:1-63. Trans. Ky. Acad. Sci. 58(2):60-66. 1997. Characterization of Tall Fescue Plantlets Regenerated from Cultured Panicle Segments/Anthers for Meiotic, Isozyme, and DNA Changes Georgia C. Eizenga USDA-ARS-MSA, Department of Agronomy, University of Kentucky, Lexington, KY 40546-0091! and Timothy D. Phillips Department of Agronomy, University of Kentucky, Lexington, KY 40546-0091 ABSTRACT Previous research identified four tall fescue (Festuca arundinacea Schreb.) genotypes (K8, K16, K25, K27) out of 20 in “Kentucky 31° that regenerated plantlets from inflorescence culture. The objectives of our study were to determine whether plantlets could be obtained from these same genotypes using anthers as the explant and to characterize the anther-derived plantlets for ploidy, meiotic, isozyme, and DNA changes. Panicle segments of each genotype were cultured on Schenk and Hildebrandt medium with 2.0 or 4.0 me/liter 2,4-dichlorophenoxyacetic acid in three separate studies. After 2 weeks, florets opened; then anthers were removed from the florets, equally divided among genotypes and hormone treatments, and cultured, 25 anthers per petri plate, on the same medium as the original panicle segment. Variables measured were callus growth, root growth, albino shoot regeneration, green shoot regeneration, and green plantlets transplanted into soil. Significantly more green plantlets regenerated from genotype K25 in two of the three studies. Thirty-two K25 and two K27 regenerants transplanted to the field were hexaploid (2n=6x=42) with normal chromosome pairing and morphologically like non-cultured tall fescue. Isozyme and RAPD profiles were identical to the maternal plants, indicating the regenerants were derived from somatic tissue, rather than being doubled haploids. INTRODUCTION Tall fescue (Festuca arundinacea Schreb., 2n=6x=42) is an open-pollinated, perennial, cool-season pasture and turf grass. Of the ca. 7.0 million acres of grassland in Kentucky, 5.5 million acres have been planted to “Kentucky 31° tall fescue (Pedersen and Lacefield 1989; Siegel et al. 1984). Kentucky 31 has poor for- age quality, and nearly 80% of the tall fescue fields in Kentucky are infected with an endo- phytic fungus (Neot yphodium coenophialum (Morgan-Jones & Gams) Glenn, Bacon, & Hanlin). A diet of infected tall fescue may be detrimental to animals. Efforts at the Univer- sity of Kentucky to improve the forage quality of tall fescue would be enhanced if non-in- fected tall fescue plants were identified that could regenerate haploid plantlets poets from anthers. ' To whom correspondence should be addressed. Present address: USDA-ARS-SPA, NRGEEC, P.O. Box 287, 2900 Hwy 130 East, Stuttgart, AR 72160. 60 Earlier research by Kasperbauer et al. (1980) reported that tall fescue haploids were obtained from inflorescence (panicle) culture; to our knowledge these plants are no longer extant. Using a similar technique, Eizenga and Dahleen (1990) identified four genotypes (plants) from 20 genotypes in non-infected Kentucky 31 tall fescue that regenerated plantlets from inflorescence culture. All 198 inflorescence-derived plantlets were morpho- logically normal. Meiotic and isozyme analyses of the 95 regenerants showed them to have 42 chromosomes, normal chromosome pairing, and the same isozyme banding patterns as the parent plants. The isozyme banding patterns suggested that the regenerants were derived from somatic tissue. Bohanec et al. (1995) used differences in RAPD (random amplified polymorphic DNA) profiles among regenerants from onion (Alli- um cepa L.) ovules to identify different ploidy levels. Also, RAPD profiles have been em- ployed to determine the phylogenetic relation- Tall Fescue Plantlets—Eizenga and Phillips 61 ships between species of Festuca and Lolium (Stammers et al. 1995). The objectives of our study were (1) to de- termine whether regeneration would occur from the four tall fescue genotypes identified by Eizenga and Dahleen (1990) using anthers as the explant and (2) to characterize the re- generated plantlets for possible ploidy, mei- otic, isozyme, and RAPD variants. MATERIALS AND METHODS The four Kentucky 31 tall fescue genotypes (K8, K16, K25, K27) we used were selected from 20 individual plants (genotypes) as _re- generating plantlets from cultured tall fescue inflorescences (Eizenga and Dahleen 1990). These genotypes were evaluated in three dif- ferent studies. Previously, the procedures used to harvest, decontaminate, and culture the panicles were reported (Eizenga and Dahleen 1990). Cul- tures of panicle segments were maintained at 25 + 2° C in the dark for 2 weeks, during which time the florets began to open. After 2 weeks the anthers were removed from the two proximal florets of each spikelet and placed on the same medium used to culture the panicle segments. Twenty-five anthers were placed on each plate. The cultured anthers were main- tained at 25 + 2°C in a 12-hour photoperiod using 40 W cool-white fluorescent lamps (ca. 20 wE/m?/s at the culture surface). Both the panicle segments and anthers were cultured on Schenk and Hildebrandt basal salt medium (Schenk and Hildebrandt 1976) with 75 g/liter sucrose, 6 g/liter agar, and 2 or 4 mg/liter 2,4-D (2, 4-dichlorophenoxyacetic acid). In the second study (trial II), kinetin (1 mg/liter) was included to determine its effect on plantlet regeneration. A total of 4800, 5400, and 9600 anthers were cultured in the three studies, trial I, trial HI, and trial III, respec- tively; these anthers were equally divided among the genotypes and hormone treat- ments. The cultured anthers were observed every 3 to 4 weeks for callus, root, albino shoot, and green shoot growth over the next 3 months. This growth was scored as the total number of anthers per plate showing evidence of callus, root, and/or shoot growth. The total number of albino and green plants regener- ated from the cultured anthers was recorded. All green plantlets were placed on a growth medium with no auxin and half-strength major salts prior to transfer to Jiffy-7 peat pellets (Jif- fy Products, Batavia, Illinois). Subsequently, the green plantlets were transplanted to the field for vernalization and to obtain inflores- cences for meiotic analyses. Data for cultured anthers were analyzed us- ing a3 X 4 X 2 factorial design with trial, genotype, and 2,4-D concentration (conc.) be- ing the factors. (Each petri plate was a unit of replication.) This analysis showed that the trial xX 2,4-D conc., genotype X 2,4-D conc., and trial X genotype X 2,4-D conc. interactions were non-significant. Only the trial < geno- type interaction was significant, so the data were reanalyzed with only the trial < geno- type interaction included to determine the least-square means. Significance was reported at the 0.05 level using a square root transfor- mation of the least-square means. Similarly, trial II data were analyzed using a 4 X 2 X 2 factorial design with genotype, 2,4-D conc., and kinetin concentration being the factors. Mitotic analyses followed the procedures of Kasperbauer et al. (1980). Meiotic analyses, pollen stainability, and enzyme assays for acid phosphatase (ACPH), glutamate oxaloacetate transaminase (GOT), malate dehydrogenase (MDH), 6-phosphogluconate dehydrogenase (6-PGD), and phosphoglucoisomerase (PGI) were according to Dahleen and Eizenga (1990). The alcohol dehydrogenase (ADH) as- say was modified according to Eizenga and Dahleen (1990). The chiasma frequency was calculated as follows: < 100. Genomic DNA was extracted from lyophi- lized ground leaf tissue with CTAB buffer (0.1 M Tris at pH 7.5, 0.7 M NaCl, 0.01 M EDTA at pH 8.0, 0.14 M B-mercaptoethanol, and 0.03 M mixed alkytrimethylammonium bro- mide [Saghai-Maroof et al. 1984]), then ex- tracted against 1 volume phenol and 1 volume of chloroform-isoamy] alcohol (24:1), followed by ethanol precipitation and a series of three ethanol rinses (15 mM ammonium acetate in 80%; 70%; 70%). The DNA pellet was dis- solved in TE (10 mM Tris at pH 8.0, 1 mM EDTA), proteinase K and 1%(w/v)SDS, and further purified with a phenol-chloroform ex- traction. RAPD analysis was modified from Williams et al. (1990) for use on a Perkin-Elmer DNA Thermocycler 480 (PE Applied Biosystems, 62 Transactions of the Kentucky Academy of Science 58(2) Figure 1. Tall fescue regenerants. Green plantlet regenerating from anther callus. Emerging roots also are visible. Foster City, CA). The reaction consisted of 25 ng sample DNA, 200 4M of each dNTP (Boehringer Mannheim, Indianapolis, IN), 37.5 ng decamer primer (Operon Technolo- gies, Inc., Alameda, CA), 1.0 U Taq polymer- ase (Boehringer Mannheim, Indianapolis, IN), and buffer (10 mM Tris pH 8.3, 50 mM KCI, 1.5 mM MgCl.) in 25.0 wL volume. The ther- mocycler was programmed for six cycles of 94°C for 1 mins 35°C for 1min:72°C for’ 2 min, followed by 35 cycles of 94°C for 1 min, 40°C for 1 min, 72°C for 2 min ending with a 6 min extension period at 72°C. Amplification DNA samples were separated by loading 12 wL of each sample, electrophoresed on 1.2% agarose gels stained with 0.06 pg/wL ethidium bromide, run with 1 X TBE buffer (89 mM Tris-base, 89 mM borate, 2 mL EDTA, pH 8.0) at 100 mV for 2 h, washed, and photo- graphed under UV light. The marker was a 1 Kb DNA Ladder (Gibco-BRL, Gaithersburg, MD). One hundred twenty-two random decamer primers were initially screened with the ma- ternal plants (K25, K27) to identify those primers that detected different polymor- phisms among the maternal plants. The fol- lowing nine primers identified differences and were selected to screen the entire set\ of 34 samples: OPES OPFI0. OB]15; OFI19; OPK19, OPTO1, OPT20, OPAM18, and OPAM19. Two independent runs were per- formed for each primer; photographs were scanned for band presence/absence in the ma- ternal plants but band absence/presence in the regenerants. RESULTS Both green (Figure 1) and albino plantlets were regenerated from cultured anthers. For- ty green plants (4 from K16, 33 from K25, 3 from K27) from three studies were regener- ated and transplanted to soil. Thirty-four (32 from K25: 2 from K27) were vernalized in the field and induced to flower. The panicles of the regenerants were morphologically like non-cultured tall fescue. Statistical analysis of the culture data indi- cated that the 2,4-D concentrations tested had no significant effect on the number of anthers producing callus, roots, and/or shoots. Similar- ly, analysis of trial II data indicated that kinetin did not significantly affect the expression of these same variables. The lack of response to kinetin was expected based on other research (Kasperbauer 1990). Genotypes K16 and K27 gave significantly more callus growth than K8 and K25 in trial I (Table 1). In trial UH, significant differences were noted in callus growth with K27 giving the most callus followed by K25 and K16. In trial I, genotype K16 regenerated signif- icantly more albino shoots (zero to 10 anthers produced shoots per petri plate) than other genotypes. In subsequent studies there were no significant differences among genotypes. The number of anthers per petri plate pro- ducing albino shoots ranged from zero to Tall Fescue Plantlets—Eizenga and Phillips 63 Table 1. Comparison of tall fescue anther culture response variables (callus growth, root regeneration, albino shoot regeneration, green shoot regeneration, green plantlets transplanted to soil) over three different studies, and four tall fescue genotypes. Response variable Experiment Genotype Callus growth Root growth Albino shoots Green shoots Green plants % anthers responding per 100 anthers Trial I K8 0.25 ab* 0.00 a* 0.25 be* 0.00 a* 0.00 a* K16 0.67 a 0.33 a alii! 0.33 a 0.33 a K25 0.00 b 0.00 a 0.00 c 0.00 a 0.00 a K27 0.67 a 0.00 a 0.75 b 0.00 a 0.00 a Trial IT K16 0.33 b 0.00 a 0.17 a 0.00 b 0.00 b K25 0.67 ab 0.00 a 0.00 a 0.44 a 0.67 a K27 0.83 a 0.17 a O.lla 0.11 ab 0.17 ab Trial II K8& 0.00 a 0.00 b 0.00 a 0.00 b 0.00 b K16 0.04 a 0.00 b 0.00 a 0.00 b 0.00 b K25 0.33 a 0.67 a 0.04 a lL13a 0.88 a K27 0.16 a 0.00 b 0.04 a 0.00 b 0.00 b * Significant at the 0.05 level using a square root transformation of the least-square means. € ae : a _ #* : 25 27 25&27 25 27 25 27 25 27 25 27 ACPH ADH GOT MDH 6-PGD_ PGI Figure 2. Tall fescue regenerants. Banding patterns of the maternal plants (K25, K27) for ACPH, ADH, GOT, MDH, 6-PGD, and PGI. Banding patterns of the regenerants were identical to those of the maternal plants, indicating that the regenerants were derived from somatic tissue rather than from microspores via spontaneous doubling. 64 Transactions of the Kentucky Academy of Science 58(2) K25 Regenerants =) M1234 | 1636 bp = Materal K27 plants Regenerants Sima: Figure 3. Tall fescue regenerants. RAPD profiles of maternal plants (K25, K27) and their respective regenerants. The identical patterns indicate that the regenerants were derived from somatic tissue rather than from microspores. Genomic DNA was amplified using the primer OPF10. K25 is distinguished from K27 by an additional band at the 1636 bp fragment. The size marker is a 1-Kb DNA ladder (Gibco-BRL). three. Mitotic analyses of the albino plantlets obtained from the shoots were unsuccessful. For green shoot regeneration, K16 ranked the highest in trial I, and K25 was the highest in trials IT and III. This ranking was the same for green plantlets transplanted to soil and nearly the same for root growth. In trial I, four green plants were regenerated from K16, but all four died when transplanted to soil in the greenhouse. In trial II, 14 of the 15 green plants transplanted to soil survived. Twelve were from K25 and two from KQ27. In trial III, 20 of the 21 green plantlets regenerated from K25 survived transplanting to soil. One of the 20 plantlets produced some green and white- striped tillers after being transplanted to soil. All 34 green plantlets that survived trans- planting to soil (32 from K25; 2 from \K27), had 42 chromosomes and normal chromo- some pairing at metaphase I. The overall mean chromosome pairing of the regenerants was 0.17 univalents, 3.03 rod bivalents, and 17.88 ring bivalents with a mean chiasma fre- quency of 92.4%. The mean pollen stainability of these same plants was 79.3%, which is with- in the range normally found in tall fescue. These meiotic analyses and pollen stainability were similar to those previously reported for the maternal plants and inflorescence-derived regenerants (Eizenga and Dahleen 1990). The maternal plants (K25, K27) had differ- ent banding patterns for five of the six en- zymes tested (Figure 2). The banding patterns were evidence of different alleles being pres- ent in the maternal plants for the isozymes ACPH-1, GOT-2, GOT-3, 6-PGD-1 and PGI-2. No variant patterns were identified among the regenerants. For the nine primers evaluated, differences in band presence/absence were noted in RAPD banding patterns between the maternal plants (K25, K27). Comparing these patterns to the regenerants, no variant patterns, band absence/presence, were found as illustrated with the OPF10 primer (Figure 3). Thus, pan- icle morphology and meiotic, isozyme, and Tall Fescue Plantlets—Eizenga and Phillips 65 RAPD analyses indicated there was no cul- ture-induced variation for these parameters. DISCUSSION The fact that isozyme and RAPD banding patterns of the regenerants were the same as the parents implies that regeneration was from somatic tissue, possibly anther wall, rather than from microspores via spontaneous dou- bling. If regeneration had been from micro- spores, some of the isozyme phenotypes should have differed from the parents similar to the segregation of isozyme markers ob- served for PGI-2 and GOT-2 in androgenetic progenies of Lolium perenne L. (Hayward et al. 1990). Also, if the plantlets had been dou- bled haploids (via spontaneous doubling of the chromosomes in the microspore), most likely a maximum of three different alleles would have been present because tall fescue is a hex- aploid species. The PGI-2 banding patterns of the K25 and K27 regenerants require four al- leles (Eizenga and Comelius 1991). In addi- tion, the tall fescue haploids obtained by Kas- perbauer et al. (1980) had fewer bands for ACPH-1 and PGI-2 than normally found in hexaploid tall fescue (Eizenga, unpublished data). Similarly, regenerants for onion ovules showed differences in RAPD profiles among regenerants of different ploidy levels, but identical profiles were found among regener- ants of the same ploidy (Bohanec et al. 1995). Screening of 764 beet (Beta vulgaris L.) re- generants for variant isozyme alleles and 60 regenerants for variant restriction fragments using RFLP (restriction fragment length poly- morphisms) analysis suggested a percentage of ca. 0.05% variant isozyme alleles and 0.1% variant restriction fragments (Sabir et al. 1992). Yamamoto et al. (1994) found that RAPD profiles could differentiate 35 rice (Oryza sativa L.) varieties, but detection of minor genetic alterations among somaclonal variants or mutants and their maternal varie- ties was not feasible. These studies suggest that neither isozyme nor random PCR tech- niques can differentiate among the 34 tall fes- cue regenerants. An additional limitation is the small sample size and the limited number of primers used. Two anther culture studies of perennial rye- grass (Lolium perenne) genotypes demonstrat- ed a significant genotypic effect on regenera- tion percentage from microspore-derived em- bryos (Madsen et al. 1995; Opsahl-Ferstad et al. 1994). Both studies suggested that regen- erability was under a relatively simple genetic control. Tall fescue is closely related to peren- nial ryegrass (Sleper 1985); thus, tissue culture regeneration in tall fescue probably is under a simple genetic control, similar to that de- scribed in perennial ryegrass. Genes for in- creasing regenerability from anther culture and green plant frequency have been mapped to chromosome arms in wheat-rye addition lines (Martinez et al. 1994) and to chromo- somal segments in maize (Zea mays L.) using RFLP markers (Armstrong et al. 1992). No significant correlation was found be- tween regenerability from cultured anthers and immature embryos of doubled haploid wheat (Triticum aestivum L.) lines, which sug- gested separate genetic control of regenera- tion (Agache et al. 1988). Similarly, lack of a significant correlation among regenerability from leaf discs, anthers, and protoplasts was reported in Solanum phureja Juz. & Buk. (Taylor and Veilleux 1992). Possibly in tall fes- cue there exists a separate genetic control for regenerability from somatic tissues and micro- spores. This would explain why only somatic tissues responded in our study. By comparison, anther culture of regener- ative lines of sorghum (Sorghum bicolor (L.) Moench.) and alfalfa (Medicago sativa L.) have been unsuccessful in producing haploid plants (Wen et al. 1991). Alternatively, as sug- gested in the studies of sorghum (Wen et al. 1991), it is possible that the tall fescue albino plantlets produced in this study were haploid. In conclusion, our research showed that se- lection for regenerability from cultured inflo- rescences was a good indicator of regenera- bility from cultured anthers. The regeneration of green plantlets apparently was from somatic tissues rather than from microspores, although it is possible that the albino plantlets were haploid. Future studies designed to obtain green tall fescue haploids will need to focus on regenerability from microspores and alter- nate methods of obtaining haploid plants. ACKNOWLEDGMENTS The authors acknowledge the excellent technical assistance of Mrs. Etta Mae Thacker and the statistical advice of Dr. Paul L. Cor- 66 Transactions of the Kentucky Academy of Science 58(2) nelius. This manuscript is Scientific Journal Series No. 96-06-114 of the Kentucky Agri- cultural Experiment Station. Mention of a trademark, vendor, or proprietary product does not constitute a guarantee or warranty of the product by the U.S. Department of Agri- culture or the University of Kentucky and does not imply its approval to the exclusion of other products that may also be suitable. LITERATURE CITED Agache, S., J. DeBuyser, Y. Henry, and J.W. Snape. 1988. Studies of the genetic relationship between anther cul- ture and somatic tissue culture abilities in wheat. PI. Breeding 100:26—33. Armstrong, C.L., J. Romero-Severson, and T.K. Hodges. 1992. Improved tissue culture response of an elite maize inbred through backcross breeding, and identi- fication of chromosomal regions important for regen- eration by RFLP analysis. Theor. Appl. Genet. $4:755— 762. Bohanec, B., M. Jakse, A. Ihan, and B. Javornik. 1995. Studies of gynogenesis in onion (Allium cepa L.): in- duction procedures and genetic analysis of regenerants. Pl. Sci. 104:215-224. Dahleen, L.S., 1990. Meiotic and iso- zymic characterization of plants regenerated from eu- and G.C. Eizenga. g ploid and selfed monosomic tall fescue embryos. Theor. Appl. Genet. 79:39-44. Eizenga, G.C., and P.L. Cornelius. 1991. Comparison of the isozyme variation in tall fescue parents and their SOULS. Euphytica 51:249-256. Eizenga, G.C., and L.S. Dahleen. 1990. Callus produc- tion, regeneration and evaluation of plants from cul- tured inflorescences of tall fescue (Festuca arundinacea Schreb.). Pl. Cell Tissue Organ Cult. 22:7-15. Hayward, M.D., A. Olesen, LK. Due, R. Jenkins, and P. Morris. 1990. Segregation of isozyme marker loci amongst androgenetic plants of Lolium perenne L. PI. Breeding 104:68—71. Kasperbauer, M.J. 1990. Haploids: derivation and evalu- ation. Pages 79-96 in M.J. Kasperbauer (ed). Biotech- nology in tall fescue improvement. CRC Press, Boca Raton, FL. Kasperbauer, M.J., R.C. Buckner, and W.D. Springer. 1980. Haploid plants by anther-panicle culture of tall fescue. Crop Sci. 20:103-106. S., A. Olesen, 1995. Inheritance of anther-culture response in peren- Madsen, B. Dennis, and S.B. Andersen. nial ryegrass (Lolium perenne L.). 165-186. Martinez, I., M. Bernard, P. Nicolas, and S. Bernard. 1994. Study of androgenetic performance and molec- ular characterization of a set of wheat-rye addition lines. Theor. Appl. Genet. 89:982-990. Opsahl-Ferstad, H.-G., A. Bjgrnstad, and O.A. Rognli. 1994. Genetic control of androgenetic response in Lol- ium perenne L. Theor. Appl. Genet. $9:133-138. Pedersen, J.F., and G.D. Lacefield. 1989. Tall fescue. Univ. Kentucky, College Agric. Coop. Ext. Serv. AGR- 59. Sabir, A., H.J. Newbury, G. Todd, J. Catty, and B.V. Ford- Lloyd. 1992. Determination of genetic stability using isozymes and RFLPs in beet plants regenerated in vitro. Theor. Appl. Genet. $4:113-117. Saghai-Maroof, M.A., K.M. Soliman, R.A. Jorgensen, and R.W. Allard. 1984. Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chro- mosomal location, and population dynamics. Proc. Natl. Acad. Sci. U.S.A. 81:8014—-8018. Schenk, R.U., and A.C. Hildebrandt. 1976. Medium and techniques for induction and growth of monocotyledon- ous and dicotyledonous plant cell cultures. Canad. J. Bot. 50:199-204. Siegel, M.R., M.C. Johnson, D.R. Varmey, 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. Sleper, D.A. 1985. Breeding tall fescue. Pl. Breeding Rev. 3:313-342. Stammers, M., J. Harris, G.M. Evans, H.D. Hayward, and J.W. Forster. 1995. Use of random PCR (RAPD) tech- nology to analyse phylogenetic relationships in the Lol- ium/Festuca complex. Heredity 74:19-27. Taylor, T.E., and R.E. Veilleux. 1992. Inheritance of com- petencies for leaf disc regeneration, anther culture, and protoplast culture in Solanum phureja and correlations among them. Pl. Cell Tissue Organ Cult. 31:95-103. Wen, F.S., E.L. Sorensen, F.L. Barnett, and G.H. Liang. 1991. Callus induction and plant regeneration from an- ther and inflorescence culture of sorghum, Euphytica 52:177-181. Williams, ].G.K., A-R. Kubelik, K.J. Livak, J.A. Rafalski, and S.V. Tingey. 1990. DNA polymorphism amplified by arbitrary primers are useful as genetic markers. Nu- cleic Acids Res. 18:6531-6535. Yamamoto, T., A. Nishikawa, and K. Oeda. 1994. DNA polymorphisms in Oryza sativa L. and Lactuca sativa L. amplified by arbitrary primed PCR. Euphytica 78: 143-148. Pl. Breeding 114: Trans. Ky. Acad. Sci. 58(2):67-73. 1997. Caddisflies (Insecta: Trichoptera) of the Mainstem of the Kentucky River, Kentucky Ronald E. Houp Kentucky Department for Environmental Protection, Division of Water, Ecological Support Section, 14 Reilly Road, Frankfort, KY 40601 and Guenter A. Schuster Department of Biological Sciences, Eastern Kentucky University, Richmond, KY 40475 ABSTRACT Collections of adult caddisflies from six Kentucky River sites were made from 1993 through 1996. Initially, stations 2 and 3 were sampled weekly in 1993-94 to determine phenology and differences in species com- position between open-river and lock-and-dam habitats. Notable differences in species composition of hy- dropsychid caddisflies prompted the addition of stations 1,4,5, and 6 in 1994. Stations 3 and 5 were open- river sites; the remaining stations were at locks and dams. Fifty species representing nine families and 23 genera were collected. Most species belonged to the families Leptoceridae (16), Hydroptilidae(12), and Hydropsychidae (10). Emergences ranged from late April through late October with peak emergences in late July and August. A total of 17 species was found at all stations, Cheumatopsyche campyla and Ceraclea tarsipunctata being the most common. The open-river stations in comparison to the lock-and-dam sites had distinctly different habitats; this was expressed as differences in species composition. Locks and dams, be- cause of the abundance of hard substrates, provided more habitat for filter feeders, scrapers, and predators. INTRODUCTION The collection, evaluation, and monitoring of aquatic communities over periods of time is a tool used to evaluate water quality and to protect aquatic life. However, biological eval- uations are of limited value when collections do not represent most or all available habitats. It has become clear that the long-term moni- toring (Kentucky Division of Water 1978— 1995) of invertebrates from the Kentucky Riv- er, using multiplate samplers, does not ade- quately represent the river's invertebrate com- munity. Some species were missed due to bi- ases of the sampler; other species, because of limitations in larval taxonomy, could be iden- tified only to the generic level. For that rea- son, light trapping was initiated to collect adult caddisflies, which in most cases can be iden- tified to the species level. Resh’s (1975) list of the caddisflies of Kentucky included records for 175 species; of these, only two were re- corded from the Kentucky River. Mozley (1979) pointed out the lack of eco- logical information regarding benthic macroin- 67 vertebrates of large rivers in North America. Prior to that time only a few studies had been conducted on these rivers. Mason et al. (1967) and Fullner (1971) studied and compared the efficiencies of various types of artificial sub- strate samplers for collection of invertebrates in large rivers. In the Ohio River, Beckett (1982) studied the phenology of Hydropsyche orris and Beckett and Miller (1982) investi- gated colonization rates of larval invertebrates on multiplate samplers in various current ve- locities. Hall (1982) compared colonization of invertebrates on basket and multiplate sam- plers in the upper Mississippi River. Beckett et al. (1983) studied distribution of larval ma- croinvertebrates from four habitat types in the lower Mississippi. Our study is the first to doc- ument the caddisfly fauna from one of the larger impounded rivers in Kentucky. KENTUCKY RIVER DRAINAGE The North, Middle, and South forks of the Kentucky River originate in the mountains of southeastern Kentucky. Each fork flows in a northwest direction, joining near the town of 68 Transactions of the Kentucky Academy of Science 58(2) - Kentucky River N (el Dix River — South Fork Kentucky River ~ : Middle Fork Kentucky River Sy North Fork Kentucky River ~ Z Figure 1. Outline of the Kentucky River drainage area showing sites of collection of Trichoptera from 1993 through 1966. Inset shows area location within Kentucky. Beattyville, to form the mainstem (Figure 1). From there, the river continues to flow north- west, joining the Ohio River at Carrollton, Kentucky, a distance of 408 km (255 miles) (Martin et. al. 1979). There are 14 locks and dams (L&D) on the mainstem, some of which have been in place for many years. For ex- ample, L&D 1 near Carrollton was completed in 1834, L&D 7 at High Bridge in 1895, and L&D 14 at Heidelburg in 1917. Locks 5-14 have been closed for over a decade. The river passes through the Cumberland, Allegheny, Table 1. ing stations along the river mainstem. Kentucky River caddisflies. Location of collect- Station Location ] Carroll Co., Lock and Dam (L&D) 1, at River Mile (RM) 4.0 (River Kilometer [Rkim] 6.4) upstream from Carrollton Jessamine Co., L&D 7, at High Bridge 3 Jessamine Co., at RM 118.6 (Rkm 191), private property 1.0 km upstream from confluence with Dix River 4 Jessamine Co., at RM 157.5 (Rkm 253), down- stream from Valley View 5 Estill Co., at RM 221.9 (Rkm 357), 2.4 km up- stream from Ravenna 6 Lee Co., at L&D 14, RM 255.0 (Rkm 410), at Heidelberg bo and Interior Plateau ecoregions (Omernik 1986). About midway along its course it flows through a part of the Interior Plateau known as the Inner Bluegrass. That segment, called the Palisades, reflects major distinctions in ge- ology and vegetation because of a geologic up- lift late in the Paleozoic Era (McFarlan 1943). Sheer limestone cliffs created by the down- ward cutting of the river are as high as 122 m (400 ft) above the river level, exposing the old- est Ordovician limestone rocks in the state (McFarlan 1943). The absence of other large streams in the Inner Bluegrass and a rapidly growing populace have greatly increased eco- nomic and recreational demands on the river. Published biological data from the Ken- tucky River are few and mostly relegated to studies of freshwater mussels that began with Rafinesque (1820). Danglade (1922) and Wil- liams (1974) documented the mussel fauna and reported on commercial potentials of the existing communities for freshwater pearls and buttons. Tolin and King (1986) surveyed the unionid mussels from L&D 4 at Frankfort, downstream to the confluence of the Ohio River. Most recently, Bradfield and Porter (1990) presented a summary of investigations related to surface water quality in the Ken- Kentucky River Caddisflies—Houp and Schuster 69 Table 2. Trichoptera collected along the Kentucky River Table 2. (continued). mainstem from 1993 through 1996. Station Station Taxa lie Ou awe oo aE its OR ee Neureclipsis crepuscularis xX xX Glossosomatidae (Walker) Protoptila maculata (Hagen) xX Polycentropus cinereus Hagen X X X Hydropsychidae Polycentropus confusus Hagen x Ceratopsyche sparna (Ross) xX Rhyacophilidae Cheumatopsyche campyla Ross =X X X X X X Rhyacophila lobifera Betten X X be carecmeyee minuscula X X X TOTAL NUMBER OF j SPECIES: 2 36 25 24 24 < Cheumatopsyche oxa Ross X ec: ATTEN dicate Cheumatopsyche pasella Ross XOX X XK X Hydropsyche betteni Ross xX X xX X Hydropsyche frisoni Ross X tucky River basin, which included sites in the Hydropsyche orris Ross X X mainstem Hydropsyche simulans Ross X xX X Potamyia flava (Hagen) Xe OX METHODS AND MATERIALS Hydroptilidae Hydroptila ajax Ross x x xX x x x Adult caddisflies were collected from six Hydroptila angusta Ross X X X X X _ stations (Figure 1) along the Kentucky River ee ar Uae XX XX mainstem from 1993 through 1996. Initially, ge cee ee nets Morton Xx Stations 2’ and 3'were sampled weekly dung Hydroptila hamata Morton X X 199394 : ee penis REA Hydroptila perdita Morton X X xX eae stations » &, O, an WETE a as m Hydroptila waubesiana Betten X X X X XK X 1994. Stations 3 and 5 represented open-river Ochrotrichia tarsalis (Hagen) X habitats; remaining stations were at L&D. Lo- Ochrotrichia xena (Ross) Xx _ . cations of all stations are shown in Table 1. Orthotrichia aegerfasciella Xe Ne xe XX , : se (Cen een Collections were made using a plastic ice One einaveristata Morton X X chest tray with about 500 ml of 70% ethanol, Oxyethira pallida (Banks) XE EX UIOXG SO XOX IX set at the base of a 12 volt Coleman lantern, Leptoceridae equipped with ultra-violet (UV) bulbs. At Ceraclea ancylus (Vorhies) X X —— ~_—__—s L&D sites, a lantern was turned on and placed Ceraclea cancellata (Betten) ee eon top of the downstream lock gate mooring Ceraclea flava (Banks) X X leageaagle Meee: dot i ft Ceraclea punctata (Banks) X X X wall at BUSK: it was turned om an hour a ct Ceraclea tarsipunctata (Vorhies) X X X X X X dark. Private docks were used at the open-riv- Ceraclea transversa (Hagen) X X X X X X __ er sites in the same manner. Species identifi- Mystacides sepulchralis (Walker) X cations were based mostly on males. Nectopsyche candida (Hagen) X X X K XK X Nectopsyche exquisita (Walker) XG Xe NN OX ae Xe RESULTS AND DISCUSSION Nectopsyche pavida (Hagen) 6 2G POS IKHOK Oecetis avara (Banks) xX This study began in early April 1993, with Oecetis cinerascens ie X X X X X X_ weekly collections at stations 2 and 3 (Figure Oecetis inconspicua (Walker) XC Xe Ne NX: il The purpose was to document species Oecetis persimilis (Banks) XG Xe XXX ). ae le d Hone B dif Triaenodes ignitus (Walker) Rex xxx x 2 CoMmpositen zal emer oT nee |e Cae ra Te Horbetariis Milne xt ferent habitat types (i.e., from a L&D site and Limnephilidae a typical open-river site). A year of weekly col- Ironoquia punctatissima X lections showed that species compositions be- (Walker) : tween stations 2 and 3 were similar (Table 1). a eg cepidaa Hagen) % However, the functional feeding requirements hilopotamidae : Chimarra obscura (Walker) ETE XE OX, XS Xe EX of some SPCCIe® reflected the types and amounts of different microhabitats between Phyrganeidae h ve eee diff, ‘ Agrypnia vestita (Walker) x the stations. On instance, dl erences 1m num- Ptilostomis ocellifera (Walker) X bers and species of hydropsychids were nota- Ptilostomis semifaciata (Say) X ble at the L&D site compared to the open- Polycentropodidae t : river site. So stations 1, 4, 5, and 6 were added Cyrnellus fraternus (Banks) SoS 2a in 1994 to increase the scope of the study. Sta- 70 Transactions of the Kentucky Academy of Science 58(2) Table 3. Seasonal occurance of trichoptera collected during this study from the Kentucky River Mainstem. Hydropsyche frisoni Hydropsyche orris Hydropsyche simulans Potamyia flava Hydroptilidae Hydroptila ajax Hydroptila angusta tion 5 was the other open-river site. Filter- feeding species (mostly hydropsychids) from all L&D sites ranged from 6 to 11 species in 1994, compared with | and 2 species at open- river sites (Table 1). Station 2 had eight of the 10 hydropsychid species. Hydropsyche orris and Potamyia flava occurred only at stations 1 and 2; Cheumatopsyche oxa, only at station 1; Ceratopsyche sparna, only at station 2; and Hydropsyche frisoni, only at station 6. Filterer species common to all stations were Chimarra obscura and Cyrnellus fraternus, though col- lected in fewer numbers at the open-river sites. Station 2 yielded three of the four spe- cies of polycentropodids, with Polycentropus confusus occurring only at station 2. Beckett and Miller (1982) observed that flow velocities contributed to different inver- tebrate communities becoming established in the Ohio River at locks and dams. In our study of adult caddisflies, it was also evident that dif- ferent sustained flow velocities were respon- sible at least in part for the diversity of filterer organisms at the L&D stations and organisms associated with the slower currents at open- river stations. Hall (1982) found 31 larval taxa in basket samplers and 21 taxa from multiplate samplers in the upper Mississippi River. In comparison, the Kentucky Division of Water's (KDOW), Biological Monitoring Program (BMP) multi- plate collections from four mainstem Ken- tucky River sites (KDOW 1978-1995) ranged from 6 to 31 larval taxa. In our study, adult caddisflies alone ranged from 24 to 36 species at six sites, for a total of 50 species throughout the river (Table 2). The augmented flow from Dix River (Her- rington Lake, hydroelectric turbines) (see Fig- ure 1) is thought to be the reason for the large number of taxa, and especially a large number of filter-feeding species, collected at station 2 (Figure 1). In addition, 14 of 16 leptocerid species were collected there. Because of the occurrences of leptocerid larvae at all stations and habitat types throughout the river, the mi- crohabitat requirements for them are appar- ently less specific in relation to flow velocities Kentucky River Caddisflies—Houp and Schuster Za Table 3. (continued). or other physical attributes. Although the numbers of individuals collected in open-river sites were much fewer for some species, there were also distributional differences. For ex- ample, Ceraclea ancylus and C. flava were col- lected only at stations 1 and 2, and Mystacides sepulchralis and Oecetis avara and were col- lected only at station 6. Five of the 12 hydroptilid species were col- lected from all stations; at station 2 nine of the 12 were common. Most of these species are scrapers during the larval stages, and the L&D provides attachment sites for them as well as for the periphyton (algae and diatoms) they consume. Ochrotrichia xena was collected only at station 2; O. tarsalis, only at station 6; and O. cristata, only at stations 3 and 5. From the distribution and numbers of hy- dropsychids, polycentropodids, and certain hy- droptilids collected, it is apparent that the L&D sites provided the microhabitats re- quired by these species. These habitats includ- ed a variety of flow regimens and firm sub- strates for needed attachment. In contrast, species such as Orthotrichia cristata and Neu- reclipsis crepuscularis, which require slower currents, were more abundant at open-river stations. It is likely that some of the species collected at light traps flew in from nearby tributary habitats. These included four species from sta- tion 1 (Ironoquia punctatissima, Ptilostomis semifaciata, and Ptilostomis ocellifera), and three from station 6 (Agrypnia vestita, Protop- tila maculata, and Rhyacophila lobifera). These species are often associated with head- water streams. The flight patterns of all species are shown in Table 3. Most caddisflies exhibit a univol- tine life cycle; our data indicate that this is true for all Kentucky River species. It is ap- parent that two different flight patterns are common among these caddisflies. The first is a single cohort that emerges over a long pe- riod of time such as in Cheumatopsyche cam- pyla or a short, synchronous emergence peri- od such as in Protoptila maculata. The second pattern is shown by species such as Ochrotri- chia tarsalis, which apparently has two cohorts emerging at different times. In addition, sea- 72 Transactions of the Kentucky Academy of Science 58(2) Table 3. (continued). Nectopsyche exquisita nneccanteesceecsseecscanceccnccasceececcescenerssscontenecsncsenssnscassenensccsscuaneusceccuerentensecs theo Nectopsyche pavida Oecetis avara Sedeg canevarrerensccmsctcnatesacennbsensstcpsceassscuscrcrd=srerscrcnntefigiccenaacstcetetsaderasessesnes! use Oecetis cinerascens cecucennnenpeacescacsecsoosucnnnccccsseusesesncscserccsussereccesssscestessusrecsessevascarsceseessesses Pes Oecetis inconspicua scnwunsendecccseveccecarcnessccsecuvsseessecessaccccsrecvesersuccssseusesseascsrcersnenensaessacscenresse= bps Oecetis persimilis Triaenodes tardus Limnephilidae Tronoquia punctatissima Pycnopsyche lepida Philopotamidae Table 3. (continued). Phyrganeidae be edesnacon OS CO ZOS CREE EREDROELERERn Cero ISRO SEC CE CEC CECE DISD CID EEOC REO ROE the Agrypnia vestita Cyrnellus fraternus Poverpesantueescaraspesceesasaccpeesnsdanstnsnnssasatstscsssssaeswustansesccaceceserscseteesuressaceses He Neureclipsis crepuscularis Rhyacophila lobifera Kentucky River Caddisflies—Houp and Schuster 73 sonal-occurrence data show that while the ma- jority of species emerge throughout the sum- mer some species, e.g., Rhyacophila lobifera, have a synchronous emergence period in the early spring and that other species, e.g., Iron- oquia punctatissima and Pycnopsyche lepida, have an emergence period restricted to a very short period in the fall. The family Leptocer- idae had the largest number of genera and species active at the same time throughout the river (Table 3). The most abundant species at all stations and throughout the study was Cheumatopsy- che campyla. The sex ratio of males to females is often skewed in light trap samples towards females. For example, we collected no males of Cheumatopsyche oxa. Oecetis inconspicua and O. persimilis were also collected as phar- ate adults attached to posterior ridges of fresh- water clams (Unionidae) in areas below station 2. It is believed that unionids represent an im- portant type of habitat for many large-river in- sect species in that they provide firm sub- strates for attachment. Locks and dams are the largest stationary objects in the Kentucky River. They, in con- cert with the various flow velocities created by these structures, provide unique habitats for filter feeders, scrapers, and predatory insects in the mainstem where such habitats are often scarce. ACKNOWLEDGMENTS We thank the following Kentucky Division of Water personnel: Scott Hankla, Terry An- derson, Dru Hawkins, Lajuanda Maybriar, Ka- thy Collins, and Karen Smathers. We acknowl- edge the help of Katy, Wes, and Laura Houp with travel and equipment. LITERATURE CITED Beckett, D.C. 1982. Phenology of Hydropsyche orris (Trichoptera: Hydropsychidae) in the Ohio River: changes in larval age structure and substrate coloniza- tion rates. Environm. Entomol. 11:1154—-1158. Beckett, D.C., and M.C. Miller. 1982. Macroinvertebrate colonization of multiplate samplers in the Ohio River: the effect of dams. Canad. J. Fish. Aquatic Sci. 39: 1622-1627. Beckett, D.C., C.R. Bingham, and L.G. Sanders. 1983. Benthic macroinvertebrates of selected habitats of the Lower Mississippi River. J. Freshwater Ecol. 2:247-261. Bradfield, A.D., and S.D. Porter. 1990. Summary of bi- ological investigations relating to surface-water quality in the Kentucky River Basin, Kentucky. U.S. Geol. Sury. Water-Resources Invest. Rep. 90-4051. Danglade, E. 1922. The Kentucky River and its mussel resources. U.S. Bur. Fish. Doc. 934:1-8. Fullner, R.W. 1971. A comparison of macroinvertebrates collected by basket and modified multiple-plate sam- plers. J. Water Pollut. Control Fed. 43:494-499. Hall, T.J. 1982. Colonizing macroinvertebrates in the Up- per Mississippi River with a comparison of basket and multiplate samplers. Freshwater Biol. 12:211-215. Kentucky Division of Water. 1978-1995. Biological Mon- itoring Program, PRI023, PRIO24, PRIO25, PRI026, 1978-1995. Frankfort, KY. McFarlan, A.C. 1943. Geology of Kentucky. University of Kentucky, Lexington, KY. Martin, W.H., W.S. Bryant, T.S. Lassetter, and J.B. Varner. 1979. The Kentucky River Palisades, flora and vegeta- tion. An inventory report to the Kentucky Chapter of the Nature Conservancy. Geographical Studies and Re- search Center, Eastern Kentucky University, Richmond, KY. Mason, W.T., J.B. Anderson, and G.E. Morrison. 1967. A limestone-filled, artificial substrate sampler-float unit for collecting macroinvertebrates in large streams. Prog. Fish-Cult. 29:74. Mason, W.T., P.A. Lewis, and T.B. Anderson. 1968. Mac- roinvertebrate collections and water quality monitoring in the Ohio River Basin 1963-1967. Coop. Rep. Office Techol. Prog., Ohio Basin Regions & Anal. Qual. Con- trol Lab. Water Qual. Office. Environm. Protect. Agen- cy, Cincinnati, OH. Mozley, S.C. 1979. Neglected characters in larval mor- phology as tools in taxonomy and phylogeny of Chiro- nomidae (Diptera). Entomol. Scand. Suppl. 10:27-36. Omernik, J.M. 1986. Aquatic ecoregions of the conter- minous United States. Ann. Assoc. Am. Geogr. 77(1): 118-125. Rafinesque, C.S. 1820. Monographie des coquilles bi- valves fluviatiles de la Riviére Ohio, contenant douze genres et soixante-huit especes. Ann. Gén. Sci. Phys. 5(13):287-322, 2 plates. Resh, V.H. 1975. A distributional study of the caddisflies of Kentucky. Trans. Kentucky Acad. Sci. 36:6-16. Tolin, W., and M. King. 1986. A survey of the freshwater mussel populations on the lower Kentucky River, Franklin, Owen, Henry and Carroll Counties, Kentucky. U.S. Fish and Wildl. Serv., Ecol. Serv., Cookeville, TN. Williams, J.C. 1974. Commercial fisheries investigations of the Kentucky River. Part I of IL: fish population studies and mussel bed surveys. Kentucky Dept. Fish and Wildlife Res., Frankfort, KY. Trans. Ky. Acad. Sci. 58(2):74-79. 1997. Noteworthy Vascular Plant Discoveries from Kentucky Gary W. Libby and Randy L. Mears Department of Biological Sciences, Eastern Kentucky University, Richmond, Kentucky 40475 and C. Tom Bloom 1209 Glade, College Station, Texas 77840 ABSTRACT Nineteen species of rare vascular plants are documented from Kentucky. Two species (Juncus validus Coville and Scirpus [Schoenoplectus] mucronatus L.) are reported as new to the state; $. mucronatus has serious potential as an invasive wetland weed. Three others (Calopogon tuberosus (L.) BSP. var. tuberosus, Polygala paucifolia Willd., and Rhynchospora macrostachya Torr. var. macrostachya) represent the only known extant Kentucky populations. New county distributional information for 14 other taxa of rare Kentucky plants is also presented. INTRODUCTION Kentucky is relatively poorly known floris- tically. Recent field studies have yielded the state records and other noteworthy collections reported here. Some of these uncommon spe- cies are restricted to specific habitats or eco- logical communities in Kentucky; others are recently naturalized or represent significant range extensions. The rarity of these species in Kentucky is based on the listing of endan- gered, threatened, special concern, and his- torical plants and animals of Kentucky (Ken- tucky State Nature Preserves Commission [KSNPC] 1996) and follows the scientific name where applicable. An asterisk preceding the scientific name indicates a non-indigenous taxon. NOTEWORTHY SPECIES Adiantum capillus-veneris L. KSNPC Threatened. LOGAN CO.: Dot 7.5 Quad., wet limestone seeps along Red River, adjacent to KY 102 (near state line), se of Keysburg, rare, 14 Sep 1995, Libby and Mears 1229 (BEREA, EKY, KNK). Significance. Cranfill (1980) stated that “no known stations are presently extant” in Kentucky. Southern maidenhair-fern has been collected in seven Kentucky counties (Adair, Boyd, Carter, Powell, Pulaski, Taylor, and Wayne) (Campbell n.d.; Cranfill 1980; McCoy 1938; Medley 1993). In addition, Cranfill 74 (1980) cited a formerly adventive colony fron Nelson County. This collection represents a westward range extension for this epipetric pteridophyte. Aureolaria patula (Chapman) Pennell. KSNPC Threatened. This species is also a can- didate (Category C2) for federal listing (Unit- ed States Fish and Wildlife Service [USFWS] 1993). ADAIR CO.: Cane Valley 7.5 Quad., along Green River Lake near Holmes, steep, rocky lakeshore, 26 Sep 1995, Libby 1277 (EKY). BARREN CO.: Meador 7.5 Quad., along Barren River Lake tailwaters near Gag- ing Station, open riverbank, 4 Oct 1995, Libby 1302 (EKY). CLINTON CoO.:: Frogue and Al- bany 7.5 Quads., along Dale Hollow Lake near Skull Bluff, steep, rocky lakeshore and edge of woods, 20 Sep 1995, Libby, Bloom, and Evans 1252 & 1253 (EKY). CUMBERLAND CoO.:: Frogue 7.5 Quad., along Dale Hollow Lake nw of Boys Island, rocky lakeshore, 6 Oct 1995, Libby 1312 (EKY). GREEN CO.: Sum- mersville 7.5 Quad., along Green River at KY 61/70 bridge, just s of Greensburg, rocky, open woods, 15 Sep 1995, Libby and Mears 1240 (EKY). TAYLOR CO.: Cane Valley 19 Quad., along Green River just below Green River Lake Dam, near KY 55 bridge, steep, forested streambanks, 26 Sep 1995, Libby 1282 (EKY). WARREN CO.:: Bowling Green South 7.5 Quad., along Drakes Creek, ca. 1/4 mile upstream from Old Scottsville Road Kentucky Vascular Plant Discoveries—Libby, Mears, and Bloom 75 bridge, steep, rocky riverbank, 14 Sep 1995, Libby and Mears 1235 (EKY). WAYNE CO.: Nevelsville 7.5 Quad., along Big South Fork Cumberland River just downstream of conflu- ence with Fanny Hollow, open, limestone bluff, 23 Aug 1995, Libby 1186 (EKY). Significance. This species was previously known from only five Kentucky counties (Ed- monson, Hart, Logan, McCreary, and Pulaski) (Braun 1941; Campbell n.d.; Evans 1989: Medley 1993). Cumberland false foxglove has now been documented in an additional eight counties (Adair, Barren, Clinton, Cumberland, Green, Taylor, Warren, and Wayne) through extensive field work (Libby 1996). These col- lections expand the range of Aureolaria patula in Kentucky and indicate that this species may not be as rare as previously thought (Kral 1983: Pennell 1935; USFWS 1993). Baptisia tinctoria (L.) R. Br. KSNPC Threatened. MCCREARY CO.: Sharp Place 7.5 Quad., growing on roadside and oak-pine upland forest margin, w side of KY 167, ca. 0.5 mile n of Kentucky/Tennessee line, 12 Jul 1995, Libby and Klein 1003 (EKY, KNK). WAYNE CoO.: Sharp Place 7.5 Quad., growing on roadside and oak-pine upland forest mar- gin, w side of KY 167, ca. 0.6 mile n of Ken- tucky/Tennessee line, 12 Jul 1995, Libby and Klein 1002 (KNK). Significance. Previously known from Letcher, Harlan, and Bell counties (Campbell n.d.; Medley 1993), yellow wild indigo has now been documented in two additional coun- ties. Bartonia virginica (L.) BSP. KSNPC Threatened. CLINTON CO.: Albany 7.5 Quad., Pyle’s Marsh, 1 mile sse of Abstons Comer, situated e of Old Burkesville Road and w of KY 1590, moss-covered areas around tree bases in swampy area, rare, 15 Jun 1995, Libby and Bloom 959 (EKY). Significance. Virginia screwstem had been collected in Calloway, Harlan, Laurel, Mc- Creary, and Whitley counties (Campbell n.d; Medley 1993). This collection represents an additional county and the first collection of this species from the Highland Rim of Ken- tucky. The more widespread B. paniculata (Michx.) Muhl. var. paniculata was found about 10 meters away in the adjacent wet woods. Gillett (1959) found that B. paniculata and B. virginica are sympatric over a large portion of their ranges and that occasional in- termediates occur. Calopogon tuberosus (L.) BSP. var. tubero- sus. KSNPC Endangered. LAUREL CO.: Vox 7.5 Quad., powerline right-of-way near Grove Recreation Area, ca. 0.7 road mile ne of large transmission line, diverse assemblage of prairie species, rare, 15 Jun 1992. No col- lection was made because of the rarity and low relative abundance at the site; photographs are on file at the Kentucky State Nature Preserves Commission, Frankfort, Kentucky. Significance. This species was formerly re- ported from 12 counties according to Camp- bell (n.d.) and Medley (1993). Prior to our re- port, however, grass pink had not been seen in Kentucky since the early 1980s (Campbell n.d.; Medley 1993). Carex crawei Dewey. KSNPC Special Concern. WAYNE CO.: Parnell 7.5 Quad., McClown Mountain Glade, ca. 0.5 air mile nw of US 90/KY 858 intersection, xeric limestone glade, locally common, 13 Jun 1995, Libby and Bloom 937 (EKY, KNK). Significance. Crawe’s sedge was described as “to be looked for” in Kentucky by Beal and Thieret (1986). Since that time, Carex crawei is known with certainty only from Bullitt, Grayson, Hardin, Larue, and Nelson counties (Campbell n.d.; Medley 1993). This collection represents a range extension and outlier from known localities in the Bluegrass and the Nashville Basin of Tennessee (Chester et al. 1993). This disjunction is probably due to the unusual occurrence of a flat, xeric limestone glade in this part of the Highland Rim. Carex stricta Lam. var. stricta. CLINTON CO.: Albany 7.5 Quad., Pyle’s Marsh, 1 mile sse of Abstons Corner, situated e of Old Bur- kesville Road and w of KY 1590, forming large tussocks in a wooded swamp, locally abundant, 15 Jun 1995, Libby and Bloom 960 (EKY, KNK). Significance. In Kentucky, this species had been collected only in Calloway, Harlan, Hart, McCreary, and Metcalfe counties (Campbell n.d.; Medley 1993). Our collection represents an additional county record for this marsh- adapted sedge. This taxon should be strongly considered as an addition to the endangered, threatened, special concern, and _ historical plants of Kentucky (KSNPC 1996). Castanea dentata (Marshall) Borkh. 76 Transactions of the Kentucky Academy of Science 58(2) KSNPC Endangered (reproducing trees only). HARLAN CoO.: Louellen 7.5 Quad., Hi-Lewis Pine Savanna, upper s-facing slopes of Hi- -Lewis Branch, near Hiram, 24 Jun 1995, Lib- by, Libby, Libby, and Evans 988 (BEREA, EKY, KNK). Significance. This collection of American chestnut in flower, including evidence of last years fruits (hulls only), indicates the presence of sexually reproducing chestnut trees in Ken- tucky. On a return visit in October 1995 we collected 15 nuts. Some of these were for- warded to J. Durrell (Kentucky Nut Growers Association) to test for reproductive potential. It is significant that 23 trees (usually multistem clumps) were observed in flower at this loca- tion. This multistem character suggests that these trees have survived previous chestnut blight [Cryphonectria parasitica (Murr.) Barr] (Hicks and Marks 1990). Cenchrus longispinus (Hack.) Fern. ES- TILL CO.: Palmer 7.5 Quad., growing in sandy riverbank downstream from Kentucky River Lock and Dam No. 11, rare, 25 Aug 1994, Libby and Walker 454 (BEREA, EKY, KNK). Significance. In Kentucky, this species had previously been reported from Ballard, Ful- ton, Hickman, Jefferson, and McCracken counties in the Coastal Plain and Bluegrass regions (Medley 1993). Our collection from the Knobs region represents a significant east- ward range extension. Cyperus croceus Vahl. MCCREARY CO.: Barthell 7.5 Quad., growing in wet area along railroad tracks adjacent to Roaring Paunch Creek near Barthell, several clumps, 7 Jul 1995, Mears and Klein s.n. (EKY). Significance. This species was recently re- ported as new to Kentucky (Madison County) (Mears and Libby 1995); ours is the second collection for the state. *Cyperus difformis L. SIMPSON CO.: wet ditch in front of gas station just w of 1-65 at exit 6 (KY 100) near Franklin, locally abun- dant, 14 Sep 1995, Libby and Mears 1219 (KNK, WKU). Significance. This naturalized species was recently reported as new to Kentucky (Mears and Libby 1995) from Knott, Floyd, and Pike counties. This collection represents an addi- tional county record and westward range ex- tension. In the United States Cyperus diffor- mis is known from Alabama, Florida, Louisi- ana, Mississippi, North Carolina, Pennsylva- nia, Tennessee, and Virginia, and also from Nebraska, Texas, and California (Bryson and Carter 1994; Godfrey and Wooten 1979; Lips- comb 1980). Godfrey and Wooten (1979) speculated that the species would likely be found elsewhere in the eastern United States. Holm et al. (1991) ranked Cyperus difformis as one of the world’s worst weeds. Bryson and Carter (1994) described it as particularly per- nicious and noted that it is spreading in the United States especially along major water- ways, through introduction at ports-of-entry, and in rice production in California. Epilobium ciliatum Raf. var. cilia- tum. MERCER CO.: Wilmore 7.5 Quad., directly below Kentucky River Lock and Dam No. 7, growing in gravel wash, rare, 21 Oct 1994, Libby and Walker 440 (EKY). Significance. This is the second report of this taxon from Kentucky. Medley (1993) cited a specimen from McCreary County collected by J. Campbell. We have seen no vouchers of this species from Kentucky. Gleason and Cronquist (1991) described the habitat and nature of this taxon as “wet places, often in unstable habitats, variable [morphology] and often somewhat weedy.” Juncus validus Coville. MCCREARY CO.: Barthell 7.5 Quad., coal mining property ca. 0.5 mile se of KY 791 just n of Blue Heron Scenic Railway, w of Coffee Branch, frequent, 5 Jul 1995, Mears and Klein s.n. (EKY). MCCREARY CoO.: Bell Farm 7.5 Quad., ca. 0.9 mile s of Pleasant Ridge Church, roadside of KY 1363, wet ditch, 13 Jul 1995, Libby, Mears, and Klein 1116B (EKY, KNK); same location, 8 Aug 1996, Libby and Libby 1403 (BEREA, EKY, KNK). MCCREARY CoO.:: Barthell 7.5 Quad., ca. 2 miles se of Bald Knob at USFS 575/USFS 6130 junction, USFS 6130 roadside, 19 Jul 1995, Libby and Mears s.n. (KNK). Significance. This species is not reported in Beal and Thieret (1986), Browne and Athey (1992), Campbell (n.d.), or Medley (1993). Our collections document the presence of Juncus validus in the Commonwealth. Lilium superbum L. KSNPC Threatened. LAUREL CO.: growing in edge of swamp along w side of U.S. 25 ca. 1 mile s of inter- section with KY 1006, 25 Jun 1993, Mears Kentucky Vascular Plant Discoveries—Libby, Mears, and Bloom We 93-234 (EKY). LAUREL CO.: e of KY 30 be- tween Raccoon and Atlanta, wet woods along Raccoon Creek, 26 Jun 1996, Libby and Har- towicz 1396 (EKY). MCCREARY CoO.: Bar- thell 7.5 Quad., floodplain along e side of South Fork Cumberland River just n of con- fluence with Bear Creek, locally frequent, 11 Jul 1995, Libby, Mears, and Klein 994 (EKY). Significance. Turk’s-cap lily has an odd distribution in Kentucky. According to Camp- bell (n.d.) and Medley (1993), this species has been collected from Big Black Mountain in Harlan County and also from wet wooded seeps in Calloway, Hickman, and McCracken counties on the Coastal Plain. According to Adams (1981) and Adams and Dress (1982), however, the Coastal Plain specimens must represent L. michiganense Farwell. Medley (1993) noted that the Coastal Plain specimens lack the “typical many-flowered inflorescence” of Lilium superbum, but the descriptions in Adams (1981), Adams and Dress (1982), and Gleason and Cronquist (1991) allow for vari- ation in number of flowers. Clearly, a thorough taxonomic review of this genus is needed in Kentucky. These represent the only known collections of this species from the Low Hills Section and Cliff Section of the Appalachian Plateau of Kentucky. *Linum usitatissimum L. MADISON CO.: Berea 7.5 Quad., growing in tire rut on roadside at the corner of Elm Street and Scaf- fold Cane Road, a European cultigen, escaped or adventive, 16 Jul 1993, Libby 343 (BEREA, EKY, KNK). Significance. This species has been col- lected in Fulton County from the Coastal Plain. A collection at the University of Ken- tucky cited by Browne and Athey (1992) and Wharton and Barbour (1991) from Fayette County may be from cultivation (Campbell n.d.; Medley 1993). Our collection represents the second report of cultivated flax naturalized in Kentucky. Grown in southeastern United States until the late 1800s (Strausbaugh and Core 1978), this European cultigen is adven- tive in fields and roadsides in the United States and southern Canada (Gleason and Cronquist 1991). Muhlenbergia cuspidata (Torr.) Rydb. KSNPC Threatened. LOGAN CO.: Dot 7.5 Quad., limestone outcrops along Red River, adjacent to KY 102 (near state line), se of Keysburg, several clumps, 14 Sep 1995, Libby and Mears 1230 (EKY). WAYNE CoO.: Mill Springs 7.5 Quad., steep, rocky cliffs at Dry Branch/Lake Cumberland confluence (w side), several large clumps on rock face, 24 Jul 1995, Libby and Bloom 1136 (EKY). Significance. In Kentucky, plains muhly had been collected from four counties (An- derson, Estill, Garrard, and Pulaski) (Bryant 1973; Campbell n.d.; Medley 1993). Our col- lections document its occurrence in two ad- ditional counties. Polygala paucifolia Willd. LAUREL CoO.: London SW 7.5 Quad., growing along bluffs on w side of Willie Green Road ca. 2.5 miles n of its intersection with KY 192, 1 May 1993, Mears 93-181 (BEREA, EKY). Significance. The report of flowering win- tergreen in Gibson (1961) was probably based on McFarland (1942): the voucher for that re- port was destroyed in the 1948 Norwood Hall fire at the University of Kentucky (Medley 1993). Our collection represents the only known extant population of this taxon in Ken- tucky. This is another taxon that should be added to the endangered, threatened, special concern, and historical plants of Kentucky (KSNPC 1996). Rhynchospora macrostachya Torr. var. ma- crostachya. KSNPC Endangered. WAYNE CO.: Parnell 7.5 Quad., Cooley’s Meadow, ca. 0.8 air mile ssw of Murl, wet meadow ne of Cooley’s tobacco barn and e of Hack Denny's comfield, rare, 27 Jul 1995, Libby and Bloom 1160 (BEREA). Significance. This is the second report of this taxon from Kentucky. The species may be extirpated from Hundred Acre Pond, Hart County (Medley 1993). Our collection may represent the only known extant population of tall beak-rush in the state. *Scirpus mucronatus L. [|=Schoenoplectus mucronatus (L.) Pallas} PULASKI CO.: Ano 7.5 Quad., pond at the end of USFS 807, w of “The Narrows” of the Rockcastle River, 26 Jul 1995, Mears and Klein s.n. (BEREA, MICH). PULASKI CO.: same location but found in two additional ponds, possibly origi- nally established from contaminated ornamen- tal water-lilies (Nymphaea sp.) planted in larg- est pond, 10 Oct 1995, Libby and Mears 1317 (BEREA, EKY, KNK, PH, WIS). Significance. Scirpus mucronatus has not 78 Transactions of the Kentucky Academy of Science 58(2) been previously reported from Kentucky (Ar- nold and Beal 1981; Beal and Thieret 1986; Browne and Athey 1992; Campbell n.d.; Med- ley 1993). This European bulrush is known from very few locations in the eastern United States (S. G. Smith, University of Wisconsin, pers. comm., 21 Oct 1996). Britton and Brown (1970) reported only one location; “a swamp in Delaware County, Pennsylvania” and fur- ther stated that S. mucronatus is widely dis- tributed in the Old World. Gleason (1952) de- scribed the distribution of S. mucronatus as “waif on ballast at Camden, New Jersey,” but Gleason and Cronguist (1991) did not list this species as occurring within northeastern Unit- ed States. This species is morphologically very similar to the native Scirpus purshianus Fern. (=Schoenoplectus purshianus (Fern.) M. T. Strong). Scirpus mucronatus can be distin- guished from S. purshianus by its sharply tri- gonal culms, trifid styles, and absence of leaf blades. Scirpus mucronatus may be estab- lished at other ponds and lakes in Pulaski County; based on the relative abundance (dominant or codominant emergent aquatic) at the known sites this bulrush should be con- sidered a pernicious weed. This taxon has been recently placed in the genus Schoeno- plectus (Bruhl 1995). ACKNOWLEDGMENTS Funds to support this research were par- tially provided by the Kentucky State Nature Preserves Commission, Kentucky River Au- thority, National Biological Service, and U.S. Fish and Wildlife Service. We thank R. Klein, S.A. Walker, E. Hartowicz, and T. Libby for valuable field assistance; Dr. R.L. Jones (Jun- cus validus), Dr. R.F.C. Naczi (Carex crawei, C. stricta var. stricta), Dr. A.A. Reznicek (Scir- pus mucronatus), Dr. A.E. Schuyler (Scirpus mucronatus), Dr. S.G. Smith (Scirpus mucron- atus), and Dr. J.W. Thieret (Baptisia tinctoria) for checking determinations; Dr. J.J.N. Camp- bell for sharing his unpublished Atlas of the Flora of Kentucky; and Dr. R.L. Thompson and the anonymous reviewers of this manu- script for critical comments. LITERATURE CITED Adams, R.M., II. graph of the turk’s-cap lilies of eastern North America 1981. A systematic study and mono- (with introduction of some new cultivars). Ph.D. dis- sertation. Cornell University, Ithaca, NY. Adams, R.M., IH, and W.J. Dress. 1982. Nodding Lilium species of eastern North America (Liliaceae). Baileya 21:165-188. Amold, S.C., and E.O. Beal. 1981. A survey of Scirpus in Kentucky with problem species-complex analyses. Trans. Kentucky Acad. Sci. 42:16-28. Beal, E.O., and J.W. Thieret. 1986. Aquatic and wetland plants of Kentucky. Kentucky Nature Preserves Com- mission, Scientific and Technical Series Number 5, Frankfort, KY. Braun, E.L. 1941. Notes on Kentucky plants V. Castanea 6:137-140. Britton, N.L., and A. Brown. 1970. An illustrated flora of the northern United States and Canada. 3 volumes. Do- ver Publications, Mineola, NY. [Reprint of a work first published in 1913] Browne, E.T., Jr., and R. Athey. 1992. Vascular plants of Kentucky: an annotated checklist. University Press of Kentucky, Lexington, KY. Bruhl, J. 1995. Sedge genera of the world: relationships and a new classification of the Cyperaceae. Austral. Syst. Bot. §:125-305. Bryant, WS. 1973. An ecological investigation of Panther Rock, Anderson County, Kentucky. Ph.D. dissertation. Southern Illinois University, Carbondale, IL. Bryson, C.T., and R. Carter. 1994. Notes on Carex, Cy- perus, and Kyllinga (Cyperaceae) in Mississippi with re- cords of eight species previously unreported to the state. Sida 16:171-182. Campbell, J.J.N. n.d. Atlas of the flora of Kentucky. Lex- ington, KY. [Unpublished draft] Chester, E.W., B.E. Wofford, R. Kral, H.R. Deselm, and A.M. Evans. 1993. Atlas of Tennessee vascular plants. Volume 1: Pteridophytes, gymnosperms, and monocots. Center for Field Biology, Austin Peay State University, Clarksville, TN. Cranfill, R. 1980. Ferns and fern allies of Kentucky. Ken- tucky Nature Preserves Commission Scientific and Technical Series Number 1, Frankfort, KY. Evans, M. 1989. A natural areas inventory of Logan, Simpson, and Warren counties, Kentucky. Tech. Rept., Kentucky State Nature Preserves Commission, Frank- fort, KY. Gibson, D. 1961. Life-forms of Kentucky flowering plants. Am. Midl. Nat. 66:1—60. Gillett, J.M. 1959. A revision of Bartonia and Obolaria (Gentianaceae). Rhodora 61:43-62. Gleason, H.A. 1952. The new Britton and Brown illus- trated flora of the northeastern United States and ad- jacent Canada. 3 volumes. Hafner Publishing Co., New York, NY. Gleason, H.A., and A. Cronquist. 1991. Manual of vas- cular plants of northeastern United States and adjacent Canada. 2nd ed. New York Botanical Garden, Bronx, NY. Godfrey, R.K., and J.W. Wooten. 1979. Aquatic and wet- Kentucky Vascular Plant Discoveries—Libby, Mears, and Bloom 79 land plants of the southeastern United States: mono- cotyledons. University of Georgia, Athens, GA. Hicks, G.F., Jr., and G.C. Marks. 1990. A survey: Amer- ican chestnut (Castanea dentata) trees in northwest In- diana. Castanea 55:291—293. Holm, L.G., D.L. Plucknett, J.V. Pancho, and J.P. Her- berger. 1991. The world’s worst weeds: distribution and biology. Krieger Publishing Company, Malabar, FL. [KSNPC] Kentucky State Nature Preserves Commission. 1996. Rare and extirpated plants and animals of Ken- tucky. Trans. Kentucky Acad. Sci. 57:69-91. Kral, R. 1983. A report on some rare, threatened, or en- dangered forest-related vascular plants of the south. Volume II. Aquifoliaceae through Asteraceae and glos- sary. Technical Publication RS-TP2. United States De- partment of Agriculture, Forest Service, Southeastern Region, Atlanta, GA. Libby, G.W. 1996. Inventory for Aureolaria patula (Chapman) Pennell (Cumberland false foxglove) in Kentucky. Pages 16-43 in D.L. White (project coordi- nator). Inventory for five candidate (Category 2) plants for federal listing. Tech. Rept., Kentucky State Nature Preserves Commission, Frankfort, KY. Lipscomb, B.L. 1980. Cyperus difformis L. (Cyperaceae) in North America. Sida §:320-327. McCoy, T.N. 1938. The ferns and fern allies of Kentucky. Am. Fern. J. 28:41-46, 101-110. McFarland, F.T. 1942. A catalogue of the vascular plants of Kentucky. Castanea 7:77—108. Mears, R.L., and G.W. Libby. 1995. New records of Cy- perus (Cyperaceae) from Kentucky. Castanea 60:79-82. Medley, M.E. 1993. An annotated catalog of the known or reported vascular flora of Kentucky. Ph.D. disserta- tion. University of Louisville, Louisville, KY. Pennell, FW. 1935. The Scrophulariaceae of eastern temperate North America. Acad. Nat. Sci. Philadelphia Monogr. 1:1—650. Strausbaugh, P.D., and E.L. Core. 1978. Flora of West Virginia. 2nd ed. Seneca Books, Morgantown, WV. [USFWS] United States Fish and Wildlife Service. 1993. Plant taxa for listing as endangered or threatened spe- cies: notice of review. Fed. Reg. 58:51144-51190. Wharton, M.E., and R.W. Barbour. 1991. Bluegrass land and life: land character, plants, and animals of the Inner Bluegrass region of Kentucky; past present and future. University Press of Kentucky, Lexington, KY. Trans. Ky. Acad. Sci. 58(2):80-84. 1997. Effects of Acidic Minesoil on Nodulation of the Legume Sericea Lespedeza (Lespedeza cuneata; Fabaceae) by Bradyrhizobium Nitrogen-fixing Bacteria Gary R. Cline and Anthony F. Silvernail Community Research Service, Kentucky State University, Frankfort, KY 40601 ABSTRACT Sericea lespedeza (Lespedeza cuneata (Dumont) G. Don; Fabaceae) is a N,-fixing legume used to revegetate minespoils in Kentucky and elsewhere. A greenhouse experiment was conducted (1) to examine the ability of sericea lespedeza to grow and form nodules in acidic minesoil and (2) to compare commercial Bradyrhizobium sp. strains with strains isolated from minespoils regarding their abilities to form nodules with lespedeza in acidic minesoil. Minesoil was adjusted to various pH levels with Al,(SO,), and CaCO,. Although growth of sericea lespedeza was significantly reduced (P = 05) by soil acidification for some inoculation treatments, the plants were able to grow relatively well under acid conditions even at a pH level of 4.2. Nodulation was significantly (P = 0.05) suppressed by increasing acidity, particularly in Al,(SO,),;-amended minesoil at pH levels of 4.2 and 4.5, which tended to affect nodulation similarly. Bradyrhizobium sp. strains responded similarly to acidified minesoil with the exception that the com- mercial mixture of strains tended to produce more numerous but smaller nodules than local strains isolated from acidic minesoil. INTRODUCTION More than 30% of the total energy and over half of the electricity produced in the United States are generated by coal burning plants (U.S. Bureau of the Census 1988). Tradition- ally, Kentucky has been one of the three lead- ing states in coal mining with more than 700,000 acres of surface-mined land having been disturbed by 4900 mining operations in the Appalachian and Eastern Interior Coal Regions of the state (Office of Surface Mining 1988a, 1988b; U.S. Bureau of the Census 1988). Revegetation of surface-mined land is necessary to reduce stream pollution, control erosion and runoff, restore minesoil produc- tivity, and improve wildlife habitats and for es- thetic reasons (Roberts et al. 1988; Vogel 1981). Minesoil acidity has been considered to be the most important soil fertility factor limiting revegetation of surface-mined land (Carpenter and Hensley 1979; Vogel 1981). Minesoils be- come acidic due to the oxidation of pyrite, a process occurring when these geologic mate- rials from lower depths are left exposed at the surface after mining. Under acidic conditions high levels of Al, Mn, and H-ions are consid- ered to be the primary sources of toxicity to plants, with Al toxicity generally considered the most important (Foy 1984). Calcium and 80 phosphorus can become deficient in soils with low pH levels and high concentrations of sol- uble Al. Nitrogen is normally deficient in newly-de- posited minesoils (Reeder and Berg 1977; Ebelhar et al. 1982), and effects of initial N fertilization are short-lived (Carpenter and Hensley 1979). Because continued fertiliza- tion is often economically and physically im- practical (Bender et al. 1985), plant produc- tion becomes dependent on N, fixation by leg- umes (Roberts et al. 1988). One of the major deterrents to achieving desirable revegetation rates on minesites is the slow establishment and growth of N,-fixing legumes, which are generally less tolerant of acidity than are non- leguminous plants (Bender et al. 1988; Vogel and Berg 1968). Sericea lespedeza, Lespedeza cuneata (Du- mont) G. Don (Fabaceae), is a warm-season, perennial, forb-legume that has been used to reclaim surface-mined lands in Kentucky. This species is not planted so frequently as it has been in the past, but it still has a place in rec- lamation as wildlife cover. Although sericea lespedeza is considered tolerant of soils with pH levels as low as 4.5 (Cline and Senwo 1993; Vogel 1981), the tolerance of its N,-fix- ing symbiosis to this pH range is unclear based upon a limited number of qualitative obser- Servicea Lespedeza and Minesoil Acidity—Cline and Silvernail 81 vations (Hyland 1938; Lyle and Evans 1979; Plass and Vogel 1973; Vogel and Berg 1968). Bacterial strains commonly used to inocu- late lespedeza were developed to fix maximum amounts of N, for a number of legume species under more favorable growing conditions than acidic minespoils. Regarding symbiotic N, fix- ation and nodulation, it was reasoned that strains isolated from sericea lespedeza growing in acidic minesoil might be more tolerant of such conditions than commercial strains. This generally proved to be true of Bradyrhizo- bium in pure culture (Cline and Senwo 1994). The objectives of our study were (1) to ex- amine the ability of sericea lespedeza to grow and form nodules in acidic minesoil and (2) to compare commercial strains with strains iso- lated from acidic minesoils regarding their abilities to form nodules in acidic minesoil. MATERIALS AND METHODS We conducted a pot experiment with four acidity treatments and four inoculation treat- ments using a completely randomized fectorial design with eight replications. Inoc ilation treatments consisted of (1) a noninoculated control, which received inorganic N, (2) a commercially available mixture of Bradyrhi- zobium sp. strains (strain numbers 4122, 3223, 32H1, 150B1, and 176A22 from Liphatech Inc., Milwaukee, WI, USA), and (3, 4) strains 41A and CRI, which were isolated from seri- cea lespedeza growing on acidic minesoils (Cline and Senwo 1994). Acidity treatments consisted of minesoil collected from an aban- doned minesite in Muhlenberg County, Ken- tucky, with a natural pH level of 5.2 (untreat- ed), minesoil adjusted to pH 6.4 with 1.6 g/kg CaCO, (Ca-msoil), and minesoil adjusted to pH 4.5 (1Al-msoil) or 4.2 (2Al-msoil) with 4.0 or 5.3 g/kg Al,(SO,);(H,O);, respectively. Minesoil was collected to a depth of 15 cm, sieved to a particle size <13 mm, and _air- dried. A previous study indicated that minesoil collected from this site was N-deficient (Cline and Senwo 1993). The minesoil was supplied with 20, 64, 32, and 5 mg/kg Ca, P, K, and Mo, respectively. Following amendment with CaCO, or Al,(SO,),, minesoil treatments were equilibrated for 8 weeks at a moisture content of 15 ml H,O/g soil. The equilibrated minesoil was mixed with river sand (50:50 by volume), and 160 g of the mixture were added to 165 ml conical containers (4 cm d X 21 cm Ray Leach Supercells, Stuewe and Sons, Inc., Cor- valis, OR). The mixture was used because les- pedeza requires well-aerated soil, and prelim- inary experiments indicated that the plant did not grow well in pure minesoil due to aeration problems associated with containers. Lespe- deza grows well in minesoils under field con- ditions. ‘Serala’ sericea lespedeza was grown from seed in a greenhouse for 20 weeks under full sunlight. Additional supplemental light (300 mol photon/m?/s photosynthetically active ra- diation) was provided with halide lamps to provide 16-hour photoperiods. Noninoculated plants received NH,NO, at rates of 50 mg N/kg minesoil after 4 weeks and 25 mg N/kg minesoil after 10 and 15 weeks. All soils were supplied with 5 mg K/kg minesoil and 1 mg Mo/kg minesoil after 10 and 15 weeks. Three milliliters of solution containing a total of 10° colony-forming units of Bradyrhizobium sp. were added to inoculated plants 2 and 4 weeks after planting. Quantification of colony-form- ing units was based upon relationships be- tween optical density (600 nm) and colony plate-counts determined for each bacterial strain including individual strains in the com- mercial mixture. At harvest, minesoil was removed from roots, and nodules were counted. Nodules and shoots were dried separately at 65° C for 48 hours prior to weighing. Minesoil from each pot was equilibrated with distilled H,O (0.6 ml H,O g/soil) for 16 hours and centrifuged at 3000 X g. Supernatants were recentrifuged at 75,000 X g and filtered using Whatman No. 42 filter paper. Final pH values were mea- sured prior to centrifugation. Toxic inorganic Al in the supernatants was determined by the pyrocatechol violet colorimetric method of Kerven et al. (1988a, 1988b); phosphorus was measured in the filtered extracts by the meth- od of Murphy and Riley (1962). Atomic ab- sorption was used to measure Mn, Mg, and Ca. Significance of main effects and interactions regarding inoculation and acidification treat- ments were determined by two-way analysis of variance. Significant differences among treat- ment means were defined by the Least Sig- nificance Difference. 82 Table 1. Mean values of acidity-related parameters (+SE, Transactions of the Kentucky Academy of Science 58(2) n = §) measured in extracts of minesoil acidity treatments used to monitor growth of sericea lespedeza and nodulation by Bradyrhizobium nitrogen-fixing bacteria. pH Acidity treatment Initial Ca-msoil 6.41 + 0.06 5.96 Untreated 5.33 + 0.03 5.15 1 Al-msoil 4.46 + 0.04 4.54 2, Al-msoil 4.18 = 0.05 4.24 RESULTS AND DISCUSSION Soil Parameters General characteristics of the minesoil have been described by Cline and Senwo (1993); parameters relevant to acidity treatments of the present study are described in Table 1. Values of initial and final pH remained rela- tively stable throughout the experiment except for Ca-msoil in which pH decreased by 0.4 units but remained near 6.0, an optimal pH for plant and bacterial growth. Concentrations of Mn and toxic Al increased with increasing minesoil acidity. Extract concentrations of P and Ca were ca. 2 uM and 5 mM, respectively, in all treatments except that extracts of un- treated minesoil contained only 1 mM Ca. Thus, negative responses of lespedeza or Bra- dyrhizobium to acidified treatments should not be related to P or Ca deficiencies. Shoot Dry Weights Shoot dry weights (above-ground plant por- tions) were generally greatest in Ca-msoil and tended to decrease with increasing acidifica- tion (Table 2). However, no chlorosis was ob- served, and shoot dry weight differences be- tween the 2Al-msoil (most acidic) and Ca- msoil (least acidic) treatments were only sig- nificant (P < 0.05) for strain 41A and the con- 2: Table Al Mn Final (uM) (uM) + 0.06 <5 6+ 0 + 0.04 Pha 1(0) 25) BE] + 0.06 85 + 3 186 + 10 + 0.06 178 + 10 SOME 7 trol. The fact that sericea lespedeza was able to produce significant yields in 2A]-msoil (pH 4.2) agrees with the findings of Cline and Sen- wo (1993) but contrasts with those of Bengt- son et al. (1973), who reported negligible les- pedeza growth at pH 4.6. In Ca-msoil, dry weight production in the N-fertilized control treatment was highest and significantly (P = 0.05) greater than production in the commer- cial and CR1 inoculation treatments. This sug- gested that sericea lespedeza inoculated with Bradyrhizobium probably did not receive suf- ficient fixed N, for maximum growth, even at the most optimal pH level (i.e., Ca-msoil). Nodulation Although large numbers of nodules were formed (Table 3), individual nodules tended to be relatively small, as indicated by their low total dry weights (Table 4). No nodules were observed on noninoculated control plants re- ceiving inorganic N. Generally nodule num- bers and dry weights from plants grown in Ca-msoil were significantly (P = 0.05) greater than numbers and weights obtained from the three more acidic soil treatments (Tables 3 and 4). Strain CR1 formed similar numbers of nodules in both untreated minesoil and Ca- msoil, but nodule weight production was sig- Mean values of sericea lespedeza mean shoot dry weight production (g + SE, n = 8) obtained in minesoil acidity treatments using different strains of Bradyrhizobium nitrogen-fixing bacteria. Values in columns followed by different letters were significantly (P < 0.05) different. Acidity treatment Commercial 41A Ca-msoil 9.94 + 0.17a 3.26 + 0. Untreated 2.96 + 0.14a 9.92 + 0. 1 Al-msoil 2.69 + 0.14a eres ax: (0), 2. Al-msoil 2.67 + 0.17a Depa a (0), Inoculation CRI Control + N 18a 2.98 + 0.16ab Soon ana! 14ab 3.30 + 0.24a 2.98 + 0.40b 17be 2.63 + 0.08b 2.26 + 0.24c 14c 2.87 + 0.16ab 2.41 + 0.1lc Servicea Lespedeza and Minesoil Acidity—Cline and Silvernail Table 3. 83 Mean values of nodule numbers (+SE, n = 8) obtained for sericea lespedeza grown in minesoil acidity treatments using different strains of Bradyrhizobium nitrogen-fixing bacteria. Values in columns followed by different letters were significantly (P = 0.05) different. OIE Sa NRE DFE OR Sep Ca UM ESA A 0 er a Inoculation Acidity treatment Commercial 41A CRI Control + N Ca-msoil 53.0 + 6.9a 71.9 + 18.3a 52.4 83a 0.0a Untreated PASO a Ht 49.3 + 6.5b 5210) Saliba 0.0a 1 Al-msoil Bont == 5.9b 27.9 + 7.0be 24.2 + 4.0b 0.0a 36.7 + 4.0b 26.5 + 4.6c 29.4 + 3.5b 0.0a 2 Al-msoil 9 ‘ nificantly (P = 0.05) higher in Ca-msoil (Table 3). Conversely, the commercial mixture of strains and strain 41A produced similar nodule dry weights in untreated minesoil and Ca- msoil but formed significantly (P = 0.05) greater numbers of nodules in Ca-msoil (Table 4). Thus, although nodulation was generally decreased in untreated minesoil compared to Ca-msoil, the response among strains differed regarding nodule numbers and size. In the more acidic 1Al-msoil and 2Al-msoil treatments, nodule dry weight production was significantly (P = 0.05) suppressed compared to other treatments (Table 4). However, no- dulation in these acidic treatments did occur, agreeing with Hyland (1938) who observed significant nodulation from pH 4.1 to 4.8. For each inoculation treatment there were no sig- nificant differences in nodule dry weights be- tween 1Al-msoil and 2Al-msoil acidification treatments, indicating that nodule dry weight production was suppressed similarly by holt treatments (pH 4.5 and 4.2, respectively). Compared to nodulation at the more optimal pH levels, nodulation in the more acidic treat- ments was suppressed less than that reported by Cline and Senwo (1993). For both 1Al- msoil and 2Al-msoil treatments, no significant differences were detected among nodule dry Table 4. Mean nodule dry weight production (mg + SE, weights obtained from the three inoculated treatments, suggesting that the strains were equally tolerant of the acid conditions. How- ever, in these treatments nodule numbers for the commercial mixture of strains tended to be significantly (P = 0.05) greater than num- bers obtained for strains 41A and CRI, indi- cating that the commercial strains tended to form more but smaller nodules under these acidic conditions. This could be important since larger nodules would be expected to contain greater amounts of nitrogenase per gram of nodule due to surface area/volume re- lationships. In previous pure culture experi- ments, strains 41A and CRI appeared to be more tolerant of acidic soil factors than com- mercial strains (Cline and Senwo 1994), whereas here they responded similarly in terms of nodule dry weight production. SUMMARY Although growth of sericea lespedeza was significantly reduced (P = 0.05) by acidifica- tion in some inoculation treatments, the plants were able to grow relatively well under the acidic conditions even in 2Al-msoil at a pH level of 4.2. Compared to Ca-msoil, nodula- tion was significantly (P = 0.05) suppressed by the more acidic treatments, particularly 1AI- n = 8) obtained for sericea lespedeza grown in minesoil acidity treatments using different strains of Bradyrhizobium nitrogen-fixing bacteria. Values in columns followed by different letters were significantly (P = 0.05) different. Inoculation Acidity treatment Commercial 41A CR1 Control + N Ca-msoil 13.9 + 2.0a 26.1 + 5.6a DAV Gy cuss Ae 0.0a Untreated 13'5y221.9a 93.3, sei 3ella 14.2 + 2.3b 0.0a 1 Al-msoil e2) eb 8.0 + 1.4b 9.4 + l.le 0.0a 2, Al-msoil 8.9 + 1.3b 10.0 = 1.2b 9.3 + 1.0c 0.0a 84 Transactions of the Kentucky Academy of Science 58(2) msoil and 2Al-msoil, which appeared to sup- press nodulation similarly for individual inoc- ulation treatments. In acidified minesoil, nod- ulation was similar among Bradyrhizobium in- oculation treatments with the exception that the commercial mixture of strains tended to produce more numerous but smaller nodules than local strains isolated from acidic mine spoils. ACKNOWLEDGMENT This research was funded by a USDA/ CSREES grant to Kentucky State University under agreement KYX-10-89-11P. LITERATURE CITED Bender, D.A., R.D. Morse, J.L. Neal, and D.D. Wolf. 1985. Delayed inoculation and starter nitrogen for en- hancing early growth and nitrogen status of Lespedeza cuneata. Pl. Soil 84:311-321. Bender, D.A., R.D. Morse, J.L. Neal, and D.D. Wolf. 1988. Field evaluation of starter N and delayed inoc- ulation of Lespedeza cuneata grown in minesoil. Pl. Soil 109: 109-113. Bengtson, G.W., S.E. Allen, D.A. Maya, and T.G. Zarger. 1973. Use of fertilizers to speed pine establishment of reclaimed coal-mine spoil in northeastern Alabama: I. Greenhouse experiments. Pages 199-225 in R.J. Hutnik and G. Davis (eds). Ecology and reclamation of dev- astated lands, Vol. 2. Pennsylvania State University Press, University Park, PA. Carpenter, P.L., and D.L. Hensley. 1979. Utilizing N,- fixing woody plant species for distressed soils and the effect of lime on survival. Bot. Gaz. 140:S76-S81. Cline, G.R., and Z. Ngewoh Senwo. 1993. Inhibitory ef- fects of acidic minesoil on the sericea lespedeza/Bra- dyrhizobium symbiotic relationship. J. Pl. Nutr. 16: 1867-1880. Cline, G.R., and Z. Ngewoh Senwo. 1994. Tolerance of lespedeza Bradyrhizobium to acidity, aluminum, and manganese in culture media containing glutamate or ammonium. Soil Biol. Biochem. 26:1067—1072. Ebelhar, M.W., R.I. Barnhisel, G.W. Akin, and J.L. Powell. 1982. Effect of lime N, P, and K amendments to sur- face-mined coal spoils. Reclamation Rev. 1:327-336. Foy, C.D. 1984. Physiological effects of hydrogen, alu- minium and manganese toxicities in acid soil. Pages 57— 97 in F. Adams (ed). Soil acidity and liming. American Society of Agronomy, Madison, WI. Hyland, H.L. 1938. Comparison of legume growth in dif- ferent soil types at varying acidity levels. J. Am. Soc. Agron. 30:111-121. Kerven, G.L., D.G. Edwards, C.J. Asher, P.S. Hallman, and S. Kokot. 1989a. Aluminum determination in soil solution. I. Evaluation of existing colorimetric and sep- aration methods for the determination of inorganic mo- nomeric aluminium in the presence of organic acid li- gands. Austral. J. Soil Res. 27:79-90. Kerven, G.L., D.G. Edwards, C.J. Asher, P.S. Hallman, and S. Kokot. 1989b. Aluminum determination in soil solution. II. short-term colorimetric procedures for the measurement of inorganic monomeric aluminium in the presence of organic acid ligands. Austral. J. Soil Res. 27:91-102. Lyle, E.S., and E.M. Evans. 1979. Revegetation of Ala- bama coal surface mines for soil cover and forage pro- duction. Reclamation Rey. 2:55-61. Murphy, J., and J.P. Riley. 1962. A modified single solu- tion method for determination of phosphate in natural waters. Anal. Chim. Acta 27:31-36. Office of Surface Mining. 198Sa. Kentucky profile. U.S. Dept. of the Interior. Washington, DC. Office of Surface Mining. 1988b. O.S.M. reclamation en- forcement annual evaluation fiscal year 1987. U.S. Dept. of the Interior. Washington, DC. Plass, W.T., and W.G. Vogel. 1973. Chemical properties and particle distribution of 39 surface-mined soils in southern West Virginia. USDA Forest Serv. Res. Paper NE-276. Reeder, J.D., and W.A. Berg. 1977. Plant uptake of in- digenous fertilizer from a Cretaceous shale and coal mine spoils. Soil Sci. Soc. Am. J. 41:919-921. Roberts, J.A., W.L. Daniels, J.C. Bell, and D.C. Martens. 1988. Tall fescue production and nutrient status on southwest Virginia mine soils. J. Environm. Qual. 17: Do=O02: U.S. Bureau of the Census. 1988. Statistical abstract of the United States. U.S. Dept. of Commerce, Washing- ton, DC. Vogel, W.G. 1981. A guide for revegetating coal mine- spoils. USDA Forest Serv. Gen. Techn. Rep. NE-68. Vogel, W.G., and W.A. Berg. 1968. Grasses and legumes for cover on acid strip-mine spoils. J. Soil Water Con- servation 23:89-91. Trans. Ky. Acad. Sci. 58(2):85-91. 1997. Structure and Composition of Three Swamp Forests on the Mississippi Alluvial Plain of Kentucky’s Jackson Purchase Region William S. Bryant Department of Biology, Thomas More College Crestview Hills, KY 41017 ABSTRACT Three swamp forests on the Mississippi alluvial plain of the Jackson Purchase Region of Kentucky are described. One forest was dominated by Taxodium distichum, one by Nyssa aquatica, and one by T. distichum and N. aquatica. In terms of basal areas and densities, these forests compared favorably to other little disturbed swamps in the southeastern United States. Basal areas of 56.5, 72.8, 84.6 m*/ha were two to three times greater than most mature upland forests of Kentucky. These swamps are remnants of a once more abundant wetland system on the Mississippi alluvial plain. INTRODUCTION Bottomland forests, like other wetland types in Kentucky and the southeastern United States, have undergone great reductions in area and changes in composition (Sharitz and Mitsch 1993). Today, only 20% of Kentucky’s originally estimated 650,000 ha of wetlands re- main; only 20% of those remnants are forested (KEQC 1992). The majority of forest rem- nants are found in the Jackson Purchase Re- gion, especially on the Mississippi alluvial plain in Ballard, Carlisle, Fulton, and Hick- man counties (KNPC 1986). Other remnants occur eastward along the Ohio, Green, and Tradewater rivers of the Western Kentucky Coalfields Region (Braun 1950; Mitsch et al. 1983). Bottomland forests, hardwoods, and deep- water swamps were first reported from the Jackson Purchase in William O. Henderson's 1820 General Land Office Survey (Bryant and Martin 1988): however, since that time little basic information on these vegetation types has been added. Aside from a few early timber surveys (DeFriese 1880, 1884; Loughridge 1888); county floras (e.g., Grubbs and Fuller 1991); a floristic list for Murphy’s Pond, a swamp in Hickman County (Adams et al. 1968); and a report on nutrient dynamics for that same swamp (King et al. 1989), the lit- erature is limited. Here and elsewhere on the Mississippi alluvial plain, few quantitative studies of bottomland forests have been re- ported (Mitsch et al. 1979). Specifically, infor- mation is scarce on the relative abundance of 85 species, structural features, gradient analysis, and diversity relations (Robertson et al. 1978). In an attempt to fill in some of the information gaps, Bryant (1991) analyzed and compared several bottomland remnants in the Jackson Purchase. Additionally, forest vegetation of the loess bluffs, which form an eastern border along the Mississippi alluvial plain, was re- ported (Bryant 1993). Forested wetlands of the Western Kentucky Coalfield have recently been the subjects of ecological analysis (Ben- son 1986; Cardamone 1985; Hill 1983; Mitsch et al. 1983, 1991; Taylor 1985) and may be better known. The purpose of this paper is to describe and quantitatively compare stand characteristics for some deepwater swamp communities in which baldcypress (Taxodium distichum) and/ or water tupelo (Nyssa aquatica) are major contributors. Comparisons to other swamp and bottomland forests are made, especially as they relate to providing answers to the forest structure and community composition. THE ENVIRONMENT The Mississippi alluvial plain is a division of the larger Coastal Plain Province (Fenneman 1938). In western Kentucky, it is the broad historic and recent floodplain of the Mississip- pi and Ohio river systems. Most of the allu- vium that covers the floodplain has been de- posited since the Pleistocene. The Mississippi alluvial plain’s aggraded surface is marked by river meanders and oxbow lakes, natural lev- ees and backswamp areas, and shallow sloughs 86 Transactions of the Kentucky Academy of Science 58(2) and marshes (Powers 1966). In their natural state the low gradient streams that flowed through the bottomlands were sluggish and widely meandering, carrying a large sediment load. As a result of microtopographical differ- ences on the floodplain, a mosaic of forested swamps and bottomland hardwoods occurs across the bottomlands. Flooding frequency, hydroperiod, and depth of flooding are factors that work with the microtopography to sort out the communities. The forested swamps occur in long, narrow, often curving sloughs, oxbow lakes, bayous, and stream margins. Lev- ees, channelization, and drainage may have modified flood plains and altered modem flooding regimes. Stands of baldcypress and water tupelo usu- ally have a long hydroperiod; however, a fall drydown of water often occurs (Wharton et al. 1982). Soils are generally saturated for most of the year. METHODS Efforts were made to locate swamp forests that showed few signs of recent human dis- turbance (e.g., stumps). I sampled three swamps during the fall drydown period in Oc- tober 1989. At that time it was possible to walk or wade into the interior of each swamp. Two of the swamps were in Ballard County (Oscar Bottoms area and Fish Lake area) and one was in Carlisle County (Back Slough area). All were in recognized wetland priority areas (KNPC 1986). Circular plots (0.04 ha), spaced at 30 m intervals, were located through the interior of each swamp. No samples were taken near the swamp edge. The number of plots sampled per swamp (or the size of the area sampled) was to equal or exceed that sampled by Anderson and White (1970) in their study of a baldcypress swamp in southern Illinois. Based on species area curves the num- ber of plots taken per stand was sufficient. Six plots were taken at Fish Lake, 10 at Back Slough, and 12 at Oscar Bottoms. All trees =10 cm at 1.4 m above the ground (dbh) were measured in each plot. Because of the swollen buttresses of baldcypress, Ander- son and White (1970) proposed measuring these trees at 2.1 m to make the basal areas more comparable to those in other forest com- munities. They found a linear relationship be- tween baldcypress diameters at 1.4 m (normal dbh) and 2.1 m. The 2.1 m dbh was about 90% (Y = .90X — 2.9) of the 1.4 m dbh. All 1.4 m dbh values for baldcypress were con- verted to the 2.1 m values based on the re- eression formula of Anderson and White (1970), and basal areas for both heights were determined. Water tupelo has swollen bases also, but no diameter conversions were. at- tempted for that species. Tree data were analyzed to relative density (RD), relative dominance (RDo), importance value (IV), and importance percentage (IP). IP, determined as IV/2, was used for compar- isons to 100. Density (trees/ha), basal area (m?/ha), species diversity (H’), and evenness (J) were determined for each swamp forest. Species diversity was calculated using the Shannon-Weaver function: H' = > (p,)(log,p;) where s is the number of species, and p, is the proportion of the total sample belonging to the ith species (Shannon and Weaver 1949). The similarity coefficient (C; Bray and Curtis 1957) was used to compare the swamps of this sample and to compare these swamps to oth- ers in the literature. The equation for calcu- lating similarity is C = (2w)/(a + b), where a equals the sum of the IVs or IPs for Swamp One, b equals the sum of the IVs or IPs for Swamp Two, and w is the sum of the lower values for the species that occur in both sam- ples. The equitability (evenness) of the stand (J) was calculated using a method based on the Shannon-Wiener function (H'): J = H’/Hmax, where Hmax is the diversity under maximum equitability (H’ = log,S), and S is the number of species in the sample. RESULTS According to the dominant species, the swamps sampled were classified as baldcypress (Back Slough), water tupelo (Fish Lake), and baldcypress-water tupelo (Oscar Bottoms). In terms of composition and stand structure (IP), the three swamps showed low degrees of sim- ilarity, C = <2% to >42% range. Similarity was lowest between the baldcypress and the Kentucky Swamp Forests—Bryant Table 1. 87 Number (N), relative density (RD), relative dominance (RDo), importance value (IV), and importance percentage (IP) for Black Slough, Carlisle County, Kentucky. N RD Taxodium distichum 17] 92.93 Planera aquatica 9 4.89 Fraxinus profunda 2 1.09 Salix nigra ] 0.54 Forestiera acuminata 1 0.54 Total 184 99.99 water tupelo communities, which were nearly monospecific stands. Of the five tree species in the baldcypress swamp (Table 1), baldcypress accounted for 93% of the density and >99% of the basal area. There were 454.5 trees/ha with a basal area of 116.5 m?/ha. The converted basal area (Anderson and White 1970) was 72.8 m?2/ha. Because of the near monospecific nature of this stand, diversity (H’ = 0.46) and evenness (J = 19.99) were low. Baldcypress knees were abundant, and fallen logs were occasional on the floor of this swamp, but no standing dead trees were observed. Cephalanthus occidentalis (buttonbush) was the most abundant shrub in the swamp. Herbs that occurred on baldcypress buttresses, fallen logs, and drydown soils included Bidens dis- coidea, Boehmeria cylindrica, H ypericum wal- teri, Lycopus rubellus, Cuscuta sp., and Sau- rurus cernuus. Anderson and White (1970) re- ported a similar but more extensive list of herbs in southern Illinois. Because the sam- pling timeframe was limited for my study, little effort was made to thoroughly characterize the herb layer of this and the other swamps. Hei- neke (1987) presented a more complete listing of plants from the middle portion of the Mis- sissippi alluvial plain. The water tupelo swamp contained only three tree species (Table 2). Water tupelo con- tributed 95% of the density and >94% of the stand basal area. There were 526.9 trees/ha. Table 2. RDo IV IP 99.05 191.98 95.99 0.30 5.19 2.60 0.07 1.16 0.58 0.54 1.08 0.54 0.03 0.57 0.29 99.99 199.98 100.00 Basal area was 85.3 m2/ha; converted basal area was 84.6 m?/ha. Species diversity (H’ = 0.36) and evenness (J = 22.71) were low in this nearly pure stand. Buttonbush was the only commonly encountered shrub. Twelve tree species were present in the baldcypress-water tupelo swamp (Table 3). Baldcypress and water tupelo accounted for 70% of the 69.9 m?/ha basal area. The con- version value was 56.5 m?/ha. There were 403.4 tress/ha; however, baldcypress and water tupelo collectively accounted for only 35% of the total density. Other species, Quercus lyrata (overcup oak), Fraxinus pennsylvanica (green ash), and Acer saccharinum (silver maple) were major associates, especially in regard to their com- bined density contributions. The other seven tree species reflected the hydric nature of the site; their presence assured a higher diversity (H’ = 2.84) relative to the other two swamps. The contributions of the several species were reflected by a greater evenness (J = 79.3). The hydroperiod may differ from that of the other two swamps sampled. Barry (1980) found that, with decreases in flooding depth and duration, more codominants appear. Average density for the three swamps sam- pled was 461.6 trees/ha; average basal area was 90.6 m?/ha; and average converted basal area was 71.3 m?/ha. Those values are com- parable to other little disturbed swamp forests in the southeastern United States (e.g., An- Number (N), relative density (RD), relative dominance (RDo), importance value (IV) and importance percentage (IP) for the Fish Lake Swamp Forest, Ballard County, Kentucky. N RD Nyssa aquatica 121 94.53 Taxodium distichum 3 2.34 Quercus lyrata 4 3.14 Total 128 100.00 RDo IV IP 94.30 188.83 94.42 1.56 3.90 1.95 4.13 7.26 3.63 99.99 199.99 100.00 88 Transactions of the Kentucky Academy of Science 58(2) Table 3. Number (N), relative density (RD), relative dominance (RDo), importance value (IV), and importance percentage (IP) for the Oscar Bottoms Swamp Forest, Ballard County, Kentucky. N RD RDo IV IP Taxodium distichum 4] 20.92 40.39 61.31 30.66 Nyssa aquatica 28 14.29 29.61 43.90 21.95 Quercus lyrata 45 22.96 11.19 34.15 17.08 Acer saccharinum Bho 16.84 5.79 22.63 RES?) Fraxinus pennsylvanica 25 12.76 6.43 19.19 9.60 Ulmus americana 8 4.08 0.40 4.48 2.24 Populus heterophylla 6 3.06 1.32 4.38 2.19 Populus deltoides 2 1.02 3.22. 4.24 2.12 Planera aquatica 4 2.04 0.12 2.16 1.08 Platanus occidentalis 2 1.02 0.92 1.94 0.97 Salix nigra ] 0.51 0.57 1.08 0.54 Forestiera acuminata ] 0.51 0.03 0.54 0.27 Total 196 100.01 99.99 200.00 100.02 derson and White 1970; Penfound 1952; Rob- ertson 1987; Schlesinger 1978). Size class distributions for baldcypress and water tupelo followed bell-curves, suggesting even-aged stands (Figures 1, 2). No trees were cored for age determinations; however, rough Number of Stems 0 30 60 90 120 Diameter Size Classes (cm.) Figure 1. Size-class distributions for baldcypress (Taxo- dium distichum) at Back Slough and for water tupelo (Nyssa aquatica) at Fish Lake in the Jackson Purchase Region of Kentucky. Gray data points = baldcypress; black data points = water tupelo. age estimates were obtained by applying av- erage diameter growth rate (3.25 mm/yr) for baldcypress at Hovey Lake, Indiana (Kozel et al. 1989) to baldcypress trees at Oscar Bot- toms and Back Slough. Maximum ages were 322 yr and 416 yr, respectively. Age estimates using Mattoon’s (1915) “rule of thumb” that 12 inches (30.5 cm) in diameter equals 100 years gave similar, but somewhat greater ages. Results of either estimate add support that these are long-established stands. DISCUSSION Baldcypress and water tupelo, either singly or in association, are clearly the dominant tree species at the hydric end of the moisture gra- dient on the Mississippi alluvial plain in Ken- tucky’s Jackson Purchase Region. Deepwater Number of Stems Diameter Size Classes (cm.) Figure 2. Size-class distribution for all trees of baldcy- press (Taxodium distichum) and water tupelo (Nyssa aquatica) in the Oscar Bottoms Swamp, Ballard County, Kentucky. Gray data points = baldcypress; black data points = water tupelo. Kentucky Swamp Forests—Bryant 89 swamps are covered by water throughout most of the year (Penfound 1952). Basal areas (m?/ ha) and densities (trees/ha) for the three swamps sampled compared favorably to other southeastern United States swamp forests (e.g., Schlesinger 1978), but basal areas greatly exceeded those reported for the Western Coalfield Region (Mitsch et al. 1991). Swamp forests tend to have significantly greater stand densities (Nelson 1997) and basal areas than most regional upland forests including such highly productive forests as the coves of the Smoky Mountains (Whittaker 1966). Basal ar- eas, converted or not, are two to three times greater than those of mature upland forests in eastern North America (Held and Winstead 1975; Martin 1992) (Table 4). Tree species di- versities are low (Robertson 1987). Mitsch et al. (1991) noted that complexity indices of cy- press-dominated systems were low. Braun (1950) observed that bottomlands from Louisiana northward to southern Illinois show a great floristic uniformity. An ecological comparison of the swamps in my study was made to a swamp in Louisiana (Conner and Day 1970) and one in southern Illinois (An- derson and White 1970). Maximum similarity values were only 58% among those locations. Bryant (1991) reported a 51% similarity be- tween a bottomland hardwood stand in Ken- tucky and one in Louisiana. Those similarities are rather low and indicate some differences in composition but especially in the contribu- tions (IVs or IPs) of species in common be- tween sampling sites along this north-south gradient. Other differences may reflect the lo- cation of sampling plots. Anderson and White (1970) reported red maple (Acer rubrum) from a southern Illinois swamp, but in western Kentucky red maple was a tree of swamp mar- gins, not of their interiors (Bryant, unpub- lished data). Guthrie (1989) also noted the ap- parent absence of water tupelo from Reelfoot Lake, Tennessee, a swamp just south of the Jackson Purchase sites. In regard to hard- woods, Bryant (1991) reported higher impor- tances for pecan (Carya illinoinensis), other hickories, and oaks in Kentucky than in bot- tomlands further south. Braun (1950) may be correct in regard to floristic similarity, but ve- getationally there appears to be a greater vari- ability in the bottomlands. Even-aged, densely stocked monospecific Table 4. Basal areas (m?/ha) for selected upland and lowland forests in Kentucky for comparison to swamps of the Jackson Purchase Region of Kentucky. Lilley Cornett Woods (Martin 1975) Sugar maple-Basswood-Tulip poplar Hemlock Chestnut oak Beech-White oak Lilley Cornett Woods (Muller 1982) Old growth composite 30.0 m?/ha 42.4 m*/ha 29.1 m?/ha 28.8 m?/ha 27.8 m2/ha Bonayer Forest (Bougher and Winstead 1974) Oak forest 31.9 m2/ha Boone County Cliffs Nature Preserve (Bryant 1978) Total forest 27.2 m/ha Lloyd Wildlife Preserve Forest (Bryant 1985) Mixed forest 35.5 m2/ha Dinsmore Woods Nature Preserve (Held and Winstead 1976) Oak-ash-maple forest Myers Woods (Held 1980) Beech forest 28.1 m?/ha 37.1 m?/ha Greenwood Forest (Chester et al. 1995) Wet woods 37.8 m?/ha Upper Bottoms (Bryant 1991) Sugarberry forest Martin Creek (Martin 1983) Southern red oak 35.6 m2/ha 29.2 m2/ha Cypress Swamp (Mitsch et al. 1991) Stagnant swamp Semi-permanent flooded 35.9 m?/ha 32.7 m2/ha stands of baldcypress or water tupelo appear to be most common in deep water (Anderson and White 1970; Johnson and Shropshire 1983). Monospecific stands of water tupelo have been attributed to past logging and re- moval of baldcypress (Robertson 1987). That was not confirmed in my study, although stumps were present in areas adjacent to sam- pling sites. Water depth and hydroperiod play extremely important roles in determining spe- cies composition on the alluvial plain. Bald- cypress-water tupelo forests may support re- cruitment of mixed bottomland species follow- ing drought or draining (Sharitz and Mitsch 1993). Swamp sites with shallower and more 90 Transactions of the Kentucky Academy of Science 58(2) variable water depths may reflect this but ar- eas where flooding is relatively constant are usually dominated by baldcypress (e.g., Back Slough) or water tupelo (e.g., Fish Lake). The low numbers of standing dead trees at the three swamps suggests that their compositions are relatively stable and not undergoing com- positional shifts as was found in the Western Kentucky Coalfield (Mitsch et al. 1991). Estimated ages of selected trees in the swamps indicate that they are remnants of long-established swamp systems. Baldcypress trees with >1.m dbh were not unusual, but none as large as the 2.3 m dbh reported by DeFriese (1880) were found. In their study of bottomland forests at Horseshoe Lake, southern Illinois, Robertson et al. (1978) found increased dominance, low- er equitability, and decrease in number of spe- cies of intermediate importance from the me- sic to the wet segment of the moisture gradi- ent. They attributed that reduction in diversity to increased flooding and poorer drainage. Wharton et al. (1982) referred to this as an anaerobic gradient. Robertson et al. (1978) stated, “The complex gradient acts as a filter which effectively screens a number of species. This filtering action continues as flooding in- creases, until swamp forests dominated by T. distichum and N. aquatica prevail.” That ap- parently is also the situation on the Mississippi alluvial plain in the Jackson Purchase Region. Ewell and Odum (1984) stated, “Under- standing wetland ecosystems is a major goal of contemporary ecological sciences.” Wetlands continue to be lost in Kentucky. It has taken over 175 years for baldcypress and water tu- pelo swamp forests to be described for the Jackson Purchase Region. More research on Kentucky's diminishing wetlands is needed so that the roles these valuable ecosystems play in the land-water interface may be better un- derstood. LITERATURE CITED Adams, B., G.E. Hunter, D.F. Austin, and K.H. Kernick. 1968. The flora of Murphy's Pond. Trans. Kentucky Acad. Sci. 29:25-28. Anderson, R.C., and J. White. 1970. A cypress swamp outlier in southern Illinois. Trans. Illinois State Acad. Sci. 63:6-13. Barry, J.M. 1980. Natural vegetation of South Carolina. Univ. South Carolina Press, Columbia, SC. Benson, K.B. 1986. Hydrologic patterns and phosphorus dynamics in forested wetlands of western Kentucky. Ph.D. dissertation, Univ. Louisville, Louisville, KY. Bougher, C.K., and J.E. Winstead. 1974. A phytosocio- logical study of a relict hardwood forest in Barren and Edmondson counties. Trans. Kentucky Acad. Sci. 35: 44-54. Braun, E.L. 1950. Deciduous forests of eastern North America. The Blakiston Company, Philadelphia, PA. Bray, J.R., and J.T. Curtis. 1957. An ordination of the upland forest communities of southern Wisconsin. Ecol. Monogr. 27:325-339. Bryant, W.S. 1978. Vegetation of the Boone County Cliffs Nature Preserve, a forest on a Kansan outwash deposit in northern Kentucky. Trans. Kentucky Acad. Sci. 39: O28 Bryant, W.S. 1985. An analysis of the Lloyd Wildlife Pre- serve Forest, Grant County, Kentucky. Trans. Kentucky Acad. Sci. 46:116—-120. Bryant, W.S. 1991. Sugarberry (Celtis laevigata) and its associates in the bottomland forests of the Jackson Pur- chase of Kentucky. Pages 93-100 in $.W. Hamilton and M.T. Finley (eds). Proceedings of the third annual sym- posium on the natural history of lower Tennessee and Cumberland river valleys. Center for Field Biology, Austin Peay State University, Clarksville, TN. Bryant, W.S. 1993. Vegetation of loess bluff ravines in the Jackson Purchase Region of Kentucky. Pages 281— 288 in A.R. Gillespie, G.R. Parker, and P.E. Pope (eds). Proceedings 9th central hardwood forest conference. USDA Forest Serv. Gen. Techn. Rep. NC-161. Bryant, W.S., and W.H. Martin. 1988. Vegetation of the Jackson Purchase of Kentucky based on the 1820 Gen- eral Land Office Survey. Pages 264-276 in D.H. Snyder (ed). Proceedings of the first annual symposium on the natural history of Lower Tennessee and Cumberland river valleys. Center for Field Biology, Austin Peay State University, Clarksville, TN. Cardamone, M.A. 1985. Wetlands and the surface mining of coal—protection and values assessment. M.S. thesis, Univ. Louisville, Louisville, KY. Chester, E.W., S.M. Noel, J.M. Baskin, C.C. Baskin, and M.L. McReynolds. 1995. A phytosociological analysis of an old-growth upland wet woods on the Pennyroyal Plain, southcentral Kentucky, USA. Nat. Areas J. 15: 297-307. Conner, W.H., and J.W. Day Jr. 1976. Productivity and composition of a baldcypress—water tupelo site and a bottomland hardwood site in a Louisiana swamp. Am. ]. Bot, 63:1354-1364. DeFriese, L.H. 1880. Report on the timbers of the dis- trict west of the Tennessee River, commonly known as the Purchase District. Geol. Surv. Kentucky n.s. 5:125— 158. DeFriese, L.H. 1884. Report on a belt of Kentucky tim- bers, extending east and west along the south-central part of the state, from Columbus to Pound Gap. Geol. Surv. Kentucky I. 5:171-232. Kentucky Swamp Forests—Bryant wall Ewell, K.C., and H.T. Odum. 1984. Introduction. Pages 1-5 in K.C. Ewel and H.T. Odum (eds). Cypress swamps. Univ. Press of Florida, Gainesville, FL. Fenneman, N.M. 1938. Physiography of eastern United States. McGraw-Hill, New York, NY. Guthrie, M. 1989. A floristic and vegetational overview of Reelfoot Lake. J. Tennessee Acad. Sci. 64:113-116. Grubbs, J.T., and M.J. Fuller. 1991. Vascular flora of Hickman County, Kentucky. Castanea 56:193-116. Heineke, T.E. 1987. The flora and plant communities of the Middle Mississippi River Valley. Ph.D. dissertation, Southern Illinois Univ., Carbondale, IL. Held, M.E. 1980. An analysis of factors related to sprout- ing and seeding in the occurrence of Fagus grandifolia Ehrh. (American beech) in the eastern deciduous forest of North America. Ph.D. dissertation, Ohio University, Athens, OH. Held, M.E., and J.E. Winstead. 1975. Basal area and cli- max status in mesic forest systems. Ann. Bot. 39:1147— 1148. Held, M.E., and J.E. Winstead. 1976. Structure and com- position of a climax forest system in Boone County, Kentucky. Trans. Kentucky Acad. Sci. 37:57-67. Hill, P.-L. 1983. Wetland-stream ecosystems of the west- em Kentucky coalfield: environmental disturbance and the shaping of aquatic community structure. Ph.D. dis- sertation, Univ. Louisville, Louisville, KY. Johnson, R.L., and FW. Shropshire. 1983. Bottomland hardwoods. Pages 175-179 in R.M. Burns (compiler). Silvicultural systems for the major forest types of the United States. USDA Forest Serv. Agric. Handb. 445. [KEQC] Kentucky Environmental Quality Commission. 1992. State of Kentucky's environment: a report of progress and problems. Frankfort, KY. King, J.M., C.D. Wilder, and J. McCandless. 1989. Sea- sonal variation in orthophosphate and inorganic nitro- gen in a western Kentucky cypress swamp (Murphy’s Pond). Trans. Kentucky Acad. Sci. 50:51—54. [KNPC] Kentucky Nature Preserves Commission. 1986. Wetland protection strategies for Kentucky. Frankfort, KY. Kozel, T.R., J. Dailey, J. Craig, and K. Welbron. 1989. Some stand characteristics of bald cypress, Taxodium distichum (L.) [sic], in an oxbow lake in extreme south- western Indiana. Trans. Kentucky Acad. Sci. 49:74-79. Loughridge, R.N. 1888. Report on the geological and economic features of the Jackson Purchase region. Ken- tucky Geol. Surv., Frankfort, KY. Martin, W.H. 1975. The Lilley Comett Woods: a stable mixed mesophytic forest in Kentucky. Bot. Gaz. 136: 171-183. Martin, W.H. 1983. Forest communities of the SRC-1 demonstration plant site: final technical report. National Technical Information Service, U.S. Dept. of Com- merce, Springfield, VA. Martin, W.H. 1992. Characteristics of old-growth mixed mesophytic forests. Nat. Areas J. 12:127-135. Mattoon, W.R. 1915. The southern cypress. USDA Bull. 972:1-74 Mitsch, W,J., J.R. Taylor, K.B. Benson, and P.L. Hill Jr. 1983. Atlas of wetlands in the principal coal surface mine region of western Kentucky. U.S. Fish and Wild- life Serv. Rep. FWS/OBS 82/72. Mitsch, W.J., J.R. Taylor, and K.B. Benson. 1991. Esti- mating primary productivity of forested wetland com- munities in different hydrologic landscapes. Landscape Ecol. 5:75-92. Muller, R.N. 1982. Vegetation patterns in the mixed mes- ophytic forest of eastern Kentucky. Ecology 63:1901— 1917. Nelson, J.C. 1997. Presettlement vegetation patterns along the 5th Principal Meridian, Missouri Territory, 1815. Am. Midl. Naturalist 137:79-94. Penfound, W.T. 1952. Souther swamps and marshes. Bot. Rev. 18:413-446. Powers, W.E. 1966. Physical geography. Appleton-Cen- tury-Crofts, New York, NY. Robertson, P.A. 1987. The woody vegetation of Little Black Slough: an undisturbed upland-swamp forest in southern Illinois. Pages 353-367 in R.L. Hay, FW. Woods, and H. DeSelm (eds). Proceedings sixth central hardwood conference, Knoxville, TN. Robertson, P.A., G.T. Weaver, and J.A. Cavanaugh. 1978. Vegetation and tree species patterns near the northern terminus of the southern floodplain forest. Ecol. Mon- ogr. 48:249-267. Schlesinger, W.H. 1978. Community structure, dynamics and nutrient cycling in the Okefenokee Cypress Swamp Forest. Ecol. Monogr. 48:43-65. Shannon, C.E., and W. Weaver. 1949. The mathematical theory of communication. Univ. Illinois Press, Urbana, IL. Sharitz, R.R., and W.J. Mitsch. 1993. Southern floodplain forests. Pages 331-372 in W.H. Martin, S.G. Boyce, and A.C. Echternacht (eds). Biodiversity of the southeastern United States: lowland forested communities. J.C. Wi- ley, New York, NY. Taylor, J.R. 1985. Community structure and primary pro- ductivity of forested wetlands in western Kentucky. Ph.D. dissertation, Univ. Louisville, Louisville, KY. Wharton, C.H., W.M. Kitchens, E.C. Pendleton, and T.W. Sipe. 1982. The ecology of bottomland hardwood swamps of the southeast: a community profile. U.S. Fish and Wildlife Serv., Biol. Serv. Prog. FWS/OBS-S81/ 37, Slidell, LA. Whittaker, R.H. 1966. Forest dimensions and production in the Great Smoky Mountains. Ecology 47:103-121. Trans. Ky. Acad. Sci. 58(2):92-95. 1997. Planning a Trip John S. Spraker, Daniel C. Biles, Mark P. Robinson Department of Mathematics Western Kentucky University Bowling Green, KY 42101 ABSTRACT The problem of minimizing the driving time for a trip is considered. A hypothetical trip from Louisville to Memphis is used as an illustration. A mathematical model is constructed and a solution technique is devised and implemented. DESCRIPTION OF THE PROBLEM We consider a hypothetical trip from Lou- isville to Memphis. We suppose the driver wishes to depart no earlier than 1100 and de- sires to arrive between 1745 and 1945, say, to attend a meeting or presentation scheduled for 2000. The driver wishes to select a depar- ture time that minimizes total travel time. We assume the driver must travel 20 miles through Louisville at a speed of 55 mph, ex- cept for lunch hour from 1200 to 1300, when speed is restricted to 45 mph, and for “rush hour” from 1600 to 1800, when speed is held to 30 mph. The motorist can then drive at 65 mph over the next 150 miles to Nashville, ex- cept for a 5-mile stretch near Elizabethtown, Kentucky, on which the speed is limited to 40 mph from 1600 to 1700 due, say, to traffic from a plant closing. The driver then travels 25 miles through the Nashville area, with the same restrictions as the Louisville area. There follows a 155-mile stretch of highway on which the motorist can proceed at 65 mph, except for a 15-mile long construction area beginning 40 miles outside of Nashville. In this area, the speed is restricted to 20 mph until 1700 after which speeds are held to 55 mph. Finally, the driver must proceed 20 miles through the Memphis area, with the same speed limits as in Louisville and Nashville. Suppose we establish a departure time of 1200. Consider Figure 1. At that departure time we are restricted to 45 mph because of lunch traffic. At 1227 we have travelled for 20 miles and reached the outskirts of Louisville, at which point we can travel at 65 mph for a 150-mile stretch. We then hit the Nashville area at 1445 where we must slow to 55 mph. Continuing in this fashion, we obtain the re- sults in Table 1. 92 RESULTS Wishing to obtain the starting time resulting in the shortest travel time, we select possible starting times spaced at regular intervals and calculate the travel time for each. Proceeding as above for each of these times would be pro- hibitively time-consuming. Therefore, we have written a computer program that calculates the arrival time and travel time for each of the possible starting times. (See Section 3 for de- tails.) We thus generate the results in Table 2. The output is restricted to those trips for which the arrival times satisfy the given con- straints. Thus we see that the optimal depar- ture time is 1300. THEORY AND IMPLEMENTATION We now consider the general problem of minimizing the driving time for a trip. One has a set of acceptable departure times S = {t,, f,, t,,...}, wheret, R is a solution to ct) =f <@) ete fe, Tl (E)) = 4a Table 1. Planning a trip. Calculation of the trip time for the trip starting at 1200. Time of day Distance travelled 1227 20 1445 170 1512 195 1549 935 1634 250 1806 350 1828 370 if and only if i) x is continuous on [f,T], ii) x’ (¢) = f(t,x(t)) for all but a finite subset of [67], iti) x(£) = Xp. Actually the definition of solution that is standard for discontinuous differential equa- tions involves Lebesgue measure theory; the interested reader can consult Chapter 2 in (Coddington and Levinson 1955). Our velocity constraint function f is assumed to satisfy the following. Definition 2: f : [t, T,) X [0,D) > R is blockwise-constant if and only if there exists a finite collection of disjoint sets A, = {(t, x) : a, =¢ =D) c= x R defined by il 15 foe ee fit, x) — 4 2 35 elsewhere This f could model a 30-mile stretch of high- way on which the speed limit is 35, except for Table 2. Planning a trip. Output from travel problem code. Starting time Arrival time Travel time 1UT2 1754 6 hr 42 min 1124 1803 6 hr 39 min 1136 1810 6 hr 34 min 1148 1817 6 hr 29 min 1200 1828 6 hr 28 min 1212 1840 6 hr 28 min 1224 1852 6 hr 28 min 1236 1858 6 hr 22 min 1248 1901 6 hr 13 min 1300 1907 6 hr 7 min 1312 1923 6 hr 11 min 1324 1942 6 hr 18 min OPTIMAL TRIP LENGTH = 6 hr 7 min 94 Transactions of the Kentucky Academy of Science 58(2) a school zone from mile 5 to mile 6 on which the speed limit is 15 from t = 4 tot = %. Before we consider numerical approaches, we must ensure that our model is well-posed. Theorem 1: Let f : [t,, T,) X [0,D) > R be blockwise-constant and positive-valued. Let f € S, t < T,. Then, one (and only one) of the following must hold: i) There exists T € (,.T,|,.x: [& T] 3-R such that x is a solution to x’(t) = f(t, x(t)) on [é,T], x) = 0 and. x(T) = D; or ii) there exists x : (&T,] — R such that x is a solution to x’(t) = f(t,x(t)) on [£T,], x(t) =.0-and x(F,)< D. The proof of Theorem | is straightforward and is left to the reader. Theorem 2: Let f : [t), T,) X [0,D) > R be blockwise-constant and positive-valued. Let t e S,t < T,. Suppose x is a function satisfying one of the two conclusions of Theorem 1. Let x be continuous, <'(f) = f(éx(t)) for all but a finite subset of the domain of < and assume there exists some f such that x(f ) = £(f). Then, x(t) = x(t) for all t in the intersection of the domains of x and x. Proof. Claim 1: x and % agree to the right of -¢, that is, 1f-¢ e.[ET)swhere © represents the right-hand endpoint of the intersection of the domains of x and «, then x(t) = x(t) for all Pelen: Proof of Claim 1: Suppose not. Then, with- out loss of generality, there exists some T € (£,T] such that <(t) > x(t). Let t* = sup{t € [t, t) | x(t) = <(t)}. Since x and & are contin- uous, we have x(t*) = x(t*) and x(t) > x(t) for all t € (t*,t]. There exists some 7 € (t*,T) such that (t,x(t)) and (t,x(t)) € interior of some A, for all t € (¢*,#). Let k, represent the constant value of f on A,. Then, for t € (t*,+), we have nee") = tk ee ei ee = x(t), a contradiction. Claim 2: x and ¥ agree to the left of t, that is, if f € (ET], where T represents the right-hand endpoint of the intersection of the domains of x and «, then x(t) = x(t) for all t € [& f]. Proof of Claim 2: Analogous to the proof of Claim 1. Corollary 1: The function x guaranteed by Theorem 1 is unique. As a result of Theorems 1 and 2, for each fe S,t < T,, the numerical procedure has one and only one solution to approximate. For each of the starting times, the numer- ical method employed in this study is an adap- tive version of Euler’s method. Given an initial value problem (IVP) Ke = fit, x), @a=t=b, x(a) =O; Euler's method, discussed in any elementary numerical analysis textbook (see, for example, (Burden and Faires 1997)) generates an ap- proximate solution to the IVP at a set of dis- erete.points T,, where @ = t).— 7 =... Ty = b, by approximating the solution x(t) locally by the first two terms in its Taylor series. Eu- ler’s method produces an approximation u, to the exact solution x(t,) at the mesh points ac- | cording to the iteration Uo ai Q, We Wat Dg rae Wn) PHIM where h, = 1, — 1;_;. Euler's method is in fact exact if the true solution x(t) is linear through- out the interval [a,b]. It is an immediate ex- tension of this fact that Euler’s method will also produce the exact answer if the exact so- lution is merely continuous and piecewise lin- ear with respect to the mesh {1,},‘,(that is, the only changes in slope occur on some subset of the set of mesh points {7},%,). In our travel problem, the function f(t,x) is blockwise con- stant, and therefore the exact solution x(t) is continuous and piecewise linear. As a conse- quence, it is possible to obtain the exact so- lution (aside from roundoff errors) from the use of Euler’s method, so long as suitable ad- justments are made. For example, a given time step may need to be modified to avoid overshooting a block. SUMMARY We have considered the problem of mini- mizing the driving time of a trip by modeling the problem with a discontinuous differential equation. Although it is straightforward (yet tedious) to solve the differential equation by hand, efficient solution of the minimization problem may be accomplished through re- peated application of Euler’s method on a Planning a Trip—Spraker, Biles, and Robinson 95 computer. We finally consider a few ideas for further work. One could include the fact that a car has maximum acceleration and deceler- ation, say, by adding the constraint x"(t) € {—d,0,c} or, x"(t) € [—d,c], where c,d are pos- itive constants. Thus, one would need to con- sider differential inclusions rather than differ- ential equations. One would now require x'(t) € [0,f(¢,x(t))] rather than x’(t) = f(t,x(t)), since with acceleration constraints one may be un- able to always drive at the speed limit. The subject of differential inclusions is not an easy one to get into; a well-known resource is (Au- bin and Cellina 1984). Also, one may wish to consider problems in which the driver may have multiple routes from which to choose. An application of graph theory may be necessary. This problem can be extended further by at- tempting to choose the ideal route and depar- ture time given certain driver preferences. For example, the driver may be interested in the best route and departure time to get from A to B, but may wish to pass through C on the way. The reader is invited to use his or her imagination to add even more twists along these lines. Finally, we mention that it would be worthwhile to consider shapes other than rectangles to subdivide the domain of f. For example, consider a school zone that drops the speed limit from 35 mph to 15 mph at 1530. In the blockwise-constant model, the driver who enters the school zone at 1529 at 35 mph will immediately realize at 1530 that he or she should slow to 15 mph as long as he or she remains in the school zone. Realistically, the warning that the speed limit has changed from 35 mph to 15 mph may only be given at the start of the school zone. In such a case, the driver mentioned above would continue at 35 mph rather than 15 mph, having no warning of the speed limit change. The reader may convince himself or herself that the proper shape in the tx-plane for this f is a trapezoid or a triangle. LITERATURE CITED Aubin, J.P., and A. Cellina. 1984. Differential inclusions. Springer-Verlag, Berlin. Burden, R.L., and J.D. Faires. 1997. Numerical analysis. 6th ed. Brooks/Cole, Pacific Grove, CA. Coddington, E.A., and N. Levinson. 1955. Theory of or- dinary differential equations. McGraw-Hill, New York, NY. Trans. Ky. Acad. Sci. 58(2):96-100. 1997. NOTES Blue Catfish (Ictalurus furcatus; Ictaluridae) Predation on the Zebra Mussel in the Ohio River near Paducah, Kentucky.—Blue catfish (Jctalurus fur- catus) were examined to determine whether they are predators of the exotic zebra mussel (Dreissena polymor- pha) in the Ohio River. Predation on zebra mussels by diving ducks, crayfish, and several fish species other than blue catfish has been described (1, 2, 3, 4). Cyprinids such as the roach (Rutilis rutilis), bull chub (Nocomis raneyi), and common carp (Cyprinus carpio) are predators of ze- bra mussels (5, 6, 7). Other fishes with molariform pha- ryngeal teeth or chewing pads—e.g., eed ear Sch (Lepomis micro- freshwater drum (Aplodinotus , grunniens ) lophus), and ealilnoutls buffalo (Ictiobus bubalus )—are considered predators of the mussels in North America (4). A common species in large rivers of southeastern Unit- ed States is the blue catfish. Blue catfish lack pharyngeal teeth but do feed on molluscs (8S). Members of the Ictal- uridae (catfish family) can feed on bivalves by swallowing them whole. French (1993) noted a zebra mussel with val On 1 Oct 1995 a commercial fisherman on the Ohio River ves intact in a brown bullhead (Ameiurus nebulosus). told us that he observed zebra mussel shells in the stom- achs of blue catfish. To confirm this observation we chose to examine blue catfish from a fish market that processed fish from the Ohio River. Blue catfish were sampled on 4, 6, and 7 1995 from Three Rivers Fish Market in Paducah, The sample consisted of 150 individuals with lengths ranging Nov Kentucky. from 25 to 50 cm. Esophagus, stomach, and intestine were removed from each fish and examined for the presence of zebra mussels. The fish had been caught in the Ohio River near Paducah between Smithland Dam and Dam 52; exact site information was not available from the commercial fishermen. A total of 1473 bivalves was recovered from 27 of the 150 catfish. Asian clam Three species of mollusks were present: (Corbicula fluminea), fawnsfoot (Truncilla donaciformis), and zebra mussel. Blue catfish with lengths of 28 to 33 cm were found to contain the majority of bivalves and consisted of 58% (87) of the sample. Of the 87 stomachs, only 31.0% (27) of individ- Eighteen stomachs 2.2% 24.4 uals of this size had fed on bivalves. ) contained zebra mussels. One stomach that had zebra mussels also had (66.7%) contained only Asian clams; six (2 three fawnsfoot mussels with lengths of 7, 11, and 12 mm. Three (11.1%) blue catfish with lengths of 29, 30, and 32 em had both Asian clams and zebra mussels. This size grouping of blue catfish had 72.6% (1040) of the total number of Asian clams and 94.6% (35) of the total number of zebra mussels. In the three species of bivalves the hinge was intact, the shells were whole, and some visceral tissue was present. A total of 1433 Asian clams was found with lengths between 1 and 12 mm. Asian clam lengths of 3 to 7 mm comprised 92.9% (1331) of the total number of Asian clams found in the stom- achs. Thirty-seven zebra mussels were collected with lengths between 3 and 15 mm. Zebra mussel lengths of 3 to 7 mm 96 comprised 78.4% (29) of the total number of zebra mussels found in stomachs. Our observations on zebra mussels in the Ohio River at Dam 52 provide a length range of 3 to 24 mm with the most common lengths of 12 to 20 mm. Our study confirms that blue catfish do feed on zebra mussels. The degree of predation and effects of this pre- dation on the Ohio River zebra mussel population are un- known. However, our findings suggest that there may be a size selection (3-7 mm) occurring when blue catfish feed on zebra mussels and Asian clams. Also, there appears to be a size range for blue catfish that feed on bivalves. Con- sidering the high numbers of blue catfish in large rivers and reservoirs in southeastern United States stad the im- pact of the zebra mussel invasion, we recommend more extensive research toward assessing predation by blue cat- fish on zebra mussels and efforts toward distinguishing whether predation is incidental or selective. We thank Hancock Biological Station of Murray State University for laboratory facilities and equipment. LITERATURE CITED. (1) Wormington, A., and J. H. Leach. 1992. Concentrations of migrant diving ducks at Point Pelee National Park, Ontario, in response to invasion of zebra mussels, Dreissena polymorpha. Canad. Field-Naturalist 106: 376-380. (2) De Leeuw, J., and M. R. Van Eerden. 1992. Size selection in diving tufted ducks Aythya fuligula explained by differential handling of small and large mussels Dreissena polymorpha. Ardea 80:353-362. (3) Love, J., and J. F. Savino. 1993. Crayfish (Orconectes virilis) predation on zebra mussels (Dreissena polymorpha). J. Freshwater Ecol. 8:253-259. (4) French, J. R. P., HI. 1993. How well can fishes prey on zebra mussels in eastern North America? Fisheries 18(6):13—19. (5) Prejs, A., K. Lewandowski, and A. Stanczykowska-Poitrowska. 1990. Size-selective predation by roach (Rutilis rutilis) on ze- bra mussel (Dreissena polymorpha): field studies. Oecologia 83:378-384. (6) Cloe, W. W,, II, G. C. Garman, and S. A. Stranko. 1993. The potential of the bull chub (Nocomis ra- neyi) as a predator of the zebra mussel (Dreissena polymor- pha) in mid-Atlantic coastal rivers. Am. Midl. Naturalist 133: 170-176. (7) Stein, R. A., J. F. Kitchell, and B. Knezevic. 1992. Selective predation by carp (Cyprinus carpio L.) on benthic molluscs in Skadar Lake, Yugoslavia. Fish Biol. 7(3): 391-399. (8) Robison, H. W., and T. M. Buchanan. 1984. Fishes of Arkansas. Univ. Arkansas Press, Fayetteville, AR — Jeffrey J. Herod, Tricia L. Frye, and James B. Sickel, Department of Biological Sciences, Murray State University, Murray, KY 42071. Rare and Extirpated Plants and Animals of Kentucky: 1997 Update.—The Kentucky State Nature Preserves Com- mission (KSNPC) published a list of rare and extirpated plants and animals of Kentucky in 1996 (1). The list, devel- oped with assistance from many scientific authorities, was based on distributional and ecological data available as of 1 Mar 1995. KSNPC (1) committed to update the list annually Notes 97 so that decision makers would have current information on rare species in Kentucky. Herein we update the 1996 list based on data available through 31 Dec 1996. The methods and status categories used herein follow KSNPC (1), with the following exceptions. The United States Fish and Wildlife Service (USFWS) (2) discontin- ued use of status review Category 2 (C2) and Category 3 (3A) to reduce confusion and to clarify that these species were not candidates for listing as endangered or threat- ened. The USFWS (2) now maintains a list of only those taxa that are candidates for listing as endangered or threatened (C, which supersedes category C1), and taxa proposed for listing as endangered or threatened (PE and PT). Those species, including three taxa subsequently list- ed as endangered by the USFWS (3), are listed in Table 1 with other species whose conservation status is being changed. Changes in nomenclature and additions to the list are presented in Tables 2 and 3, respectively. Common names are used only when there is a nomenclature change Table 1. Old Plants Aureolaria patula Berchemia scandens Elodea nuttallii Halesia tetraptera Lonicera dioica var. orientalis Trepocarpus aethusae Triplasis purpurea Viburnum lentago DAAC Des Snails Anguispira rugoderma S Mussels Alasmidonta atropurpurea = Epioblasma brevidens — Epioblasma capsaeformis — Crustaceans Cambarus ornatus S Orconectes inermis AR Insects Pseudanophthalmus catoryctos Pseudanophthalmus pholeter Fishes Ammocrypta clara X Etheostoma cinereum T Etheostoma tippecanoe S Macrhybopsis gelida = Macrhybopsis meeki Percina evides Birds Aquila chrysaetos X from KSNPC (1), to identify undescribed species, and when a species is added to the list. Sources for plant names are Gleason and Cronquist (4), McAtee (5), and Wilbur (6). Sources consulted for animal names are as follows: gastropods—Hubricht (7) and Turgeon et al. (8); insects—Barr (9), Garrison (10), and The Dragonfly So- ciety of the Americas (11); fishes—Robbins et al. (12) and Warren, Burr, and Grady (13); and birds—American Or- nithologists’ Union (14). We welcome any questions or comments about this update or KSNPC (1). We thank the following individuals for sharing infor- mation and for their assistance: W.M. Andrews, Kentucky Division of Environmental Services; T.C. Barr Jr, Uni- versity of Kentucky; D.L. Batch, J. Kiser, G. Libby, G.A. Schuster, and T. Weckman, Eastern Kentucky University; H.D. Bryan, Eco-Tech, Inc.; B.M. Burr, Southern Illinois University at Carbondale; S.M. Call and K.L. Smathers, Kentucky Division of Water; C. Cook, Florida State Col- lection of Arthropods; C.V. Covell Jr., and W.D. Pearson, Conservation status changes for rare and extirpated Kentucky plants and animals. KSNPC! US? New Old New S oes a T wt soo T sans wa T pL ae E ee ales T Lins atk, H = ae delist — — T es et = PE E — PE E — PE E delist — — S ben a E es pis E ne ce E we aay S can iss delist — — = GI GC = Cl C delist — -— delist —_ — '§ = Special Concem, T = Threatened, E = Endangered, H = Historic, X = Extirpated or Extinct. 2 PE = Proposed Endangered, E = Endangered, C = Candidate for Listing, Cl = Candidate for Listing. 98 Transactions of the Kentucky Academy of Science 58(2) Table 2. Nomenclature changes for rare and extirpated Kentucky plants and animals. Old name New name Plants Carex atlantica ssp. capillacea Howe sedge Gentiana alba Halesia tetrapera var. tetraptera Heracleum maximum Insects Pseudanophthalmus abditus A cave beetle Pseudanophthalmus hypolithos Stone-dwelling cave beetle Pseudanophthalmus inexpectatus A cave beetle Pseudanophthalmus scholasticus Schoolhouse cave beetle Fishes Etheostoma clarum Etheostoma pellucidum Etheostoma vivax Notropis sp. Palezone shiner Birds Elanoides forficatus forficatus American swallow-tailed kite Casmerodius albus Prickly bog sedge Gentiana flavida Halesia tetraptera Heracleum lanatum Concealed cave beetle Ashcamp cave beetle Surprising cave beetle Scholarly cave beetle Ammocrypta clara Ammocrypta pellucida Ammocrypta vivax Notropis albizonatus Swallow-tailed kite Ardea alba University of Louisville; D. Dourson and J.R. MacGregor, United States Forest Service; J.J. Lewis; L.E. Kornman, B.D. Laflin, and K.W. Prather, Kentucky Department of Fish and Wildlife Resources; T.L. Poulson, University of Illinois-Chicago; S.P. Rice, Kentucky Transportation Cab- Table 3. tucky plants and animals. Additions to the list of rare and extirpated Ken- KSNPC'! US? status status Plants Viburnum molle Missouri arrow-wood IE — Snails Helicodiscus notius specus A snail 0 att Helicodiscus punctatellus Punctate coil S — Insects Celithemis verna Double-ringed pennant S — Fishes Etheostoma percnurum Duskytail darter E E Special Concern, T = Threatened, E = Endangered. IS=S§ 2>E = Endangered. inet; J. Slapcinsky, Field Museum of Natural History; C.A. Taylor, Illinois Natural History Survey; and K.J. Tennes- sen. LITERATURE CITED. (1) Kentucky State Nature Preserves Commission. 1996. Rare and extirpated plants and animals of Kentucky. Trans. Kentucky Acad. Sci. 57: 69-91. (2) U.S. Fish and Wildlife Service. 1996. Endan- gered and threatened wildlife and plants; review of plant and animal taxa that are candidates for listing as endan- gered or threatened species; notice of review. Fed. Reg. 61:7596-7613. (3) United States Fish and Wildlife Ser- vice. 1997. Endangered and threatened wildlife and plants; determination of endangered status for the Cum- berland elktoe, oyster mussel, Cumberlandian combshell, purple bean, and rough rabbitsfoot. Fed. Reg. 62:1647— 1658. (4) Gleason, H.A., and A. Cronquist. 1991. Manual of vascular plants of northeastern United States and ad- jacent Canada. 2nd ed. The New York Botanical Garden, Bronx, NY. (5) McAtee, W.L. 1956. A review of the nearc- tic Viburnum. W.L. McAtee, Chapel Hill, NC. (6) Wilbur, R.L. 1988. The correct scientific name of the pale, yellow or white gentian of the eastern United States. Sida 13: 161-165. (7) Hubricht, L. 1985. The distributions of the native land mollusks of the eastern United States. Field- iana Zool. 24:1-191. (8) 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 inverte- Notes 99 brates from the United States and Canada: mollusks. Am. Fish. Soc. Spec. Publ. 16. (9) Barr, T.C., Jr. 1996. Cave beetle status survey and prelisting recovery project. Re- port prepared for the U.S. Fish and Wildlife Service and the Kentucky Dept Fish Wildl. Resources, Frankfort, KY. (10) Garrison, R.W. 1991. A synonymic list of the New World Odonata. Argia 3(2):1-30. (11) The Dragonfly So- ciety of the Americas. 1996. Common names of North American dragonflies and damselflies. Argia 8(2). (12) Robbins, C.R., R.M. Bailey, C.E. Bond, J.R. Brooker, E.A. Lachner, R.N. Lea, and W.B. Scott. 1991. Common and scientific names of fishes from the United States and Can- ada. 5th ed. Am. Fish. Soc. Spec. Publ. 20. (13) Warren, M.L., Jr, B.M. Burr, and J.M. Grady. 1994. Notropis al- bizonatus, a new cyprinid fish endemic to the Tennessee and Cumberland River drainages, with a phylogeny of the Notropis procne species group. Copeia 1994:S68—886. (14) American Omithologists’ Union. 1995. Fortieth supple- ment to the American Ornithologists’ Union check-list of North American birds. Auk 112:819-830.—Kentucky State Nature Preserves Commission, 801 Schenkel Lane, Frankfort, KY 40601. Epilobium brachycarpum (Onagraceae) in Ken- tucky.—While collecting plants in a railroad yard in northern Kentucky, we noted, among Hypericum perfor- atum and Potentilla recta, a number of non-flowering in- dividuals of a plant we did not recognize. We eventually concluded that it was probably Epilobium brachycarpum C. Pres] (E. paniculatum Nutt.) (Figure 1), a xerophytic annual new to the flora of Kentucky. Our conclusion was proved correct when, later, the plants came into flower. In the conterminous U.S., the plant was known east of the Mississippi only from Minnesota and Wisconsin. Further west, it occurs in the Dakotas and from Montana south to New Mexico and west to the Pacific; in Canada it is known from Quebec to British Columbia (International Vascular Plant Database, Biota of North America Pro- gram, North Carolina Botanical Garden, University of North Carolina, Chapel Hill, NC 27599). The species is distinctive even in non-flowering condi- tion, with (1) its elongate leaves that often have fascicles of smaller leaves in their axils and are folded up along the midrib (V-shaped), sharply acute, and entire to obscurely toothed, the teeth low, gland-tipped, and mostly 1 to 4 mm apart; and (2) its cracking and exfoliating epidermis at the base of the stem in older plants. Its flowers have pink petals about 6 mm long that are lobed to about the middle; the pollen is shed as monads, not as tetrads as in most species of Epilobium (1). Our plants are glabrous. Although infra- specific taxa have been described for this polymorphic spe- cies, none of these was recognized by Raven (1). We cite here a voucher specimen for the presence of E. brachycarpum in Kentucky: KENTUCKY. Kenton Co., in gravelly soil along tracks in railroad yard along Licking River ca. % mile s of 1-275 bridge, ca. 2 miles s of Latonia, 23 Jul 1997, Thieret and Allen 60239 (KNK, MO). Figure 1. Epilobium brachycarpum. Photocopy of pressed specimens, X ¥%. LITERATURE CITED. (1) Raven, P.H. 1976. Generic and sectional delimitation in Onagraceae, tribe Epilo- bieae. Ann. Missouri Bot. Gard. 63:326-340.—John T. Kartesz, Biota of North America Program, North Caro- lina Botanical Garden, University of North Carolina, Chapel Hill, NC 27599; Peter Allen and John W. Thier- et, Department of Biological Sciences, Norther Ken- tucky University, Highland Heights, KY 41099. Horned-Pondweed, Zannichellia palustris (Zanni- chelliaceae) from Northern and Central Kentucky.— Zannichellia, a genus of aquatic monocots of the family Zannichelliaceae, is represented in North America by a sin- gle species, Z. palustris L. (Figure 1). Beal and Thieret (1) located no Kentucky voucher specimens for the species but indicated that it is “possibly to be expected” in the state. They also noted that a population of this species formerly occurred just across the Ohio River from northem Ken- tucky in Hamilton County, Ohio, but was destroyed by ex- pansion of the city of Cincinnati. In 1987 Patricia Haragan found the species in Fayette County but did not report the find. Two years later Webb and Chester (2) published Ken- tucky records of Zannichellia for Calloway, Lyon, Marshall, and Trigg counties (Kentucky Lake). In May 1994, while looking for plants along a proposed 100 Figure 1. achenes (X6). Drawing by John Myers; used by permis- sion of Flora of North America. Zannichellia palustris. Plant (<%); lower right, right-of-way for the reconstruction of KY 17, near Inde- pendence, Kenton County, Kentucky, we found horned- pondweed in a pond with Eleocharis obtusa and Lemna minor; in May 1995 we found Zannichellia at two sites in Bourbon County (in a roadside pool along US 68, ca. 0.5 miles northeast of Paris, and in a pond north of Millers- burg); and in July 1996 we discovered a population in a pond off KY 356, ca. 2 miles east of Cynthiana, Harrison County. The documented Kentucky distribution of this Transactions of the Kentucky Academy of Science 58(2) Figure 2. Zanichellia palustris. Documented county dis- tribution in Kentucky. species now includes the northern and central parts of the state (Figure 2). Additional field work will probably yield more Kentucky records. Our voucher specimens are as follows: KENTUCKY. Bourbon Co., US 68, ca. 0.5 miles ne of Paris, roadside pool, 22 May 1995, Meade 2169 (MOKY, Morehead State University Herbarium); Bourbon Co., n end of Millers- burg, farm pond, 23 May 1995, Meade 2229 (MOKY, Morehead State University Herbarium); Fayette Co., 8 miles e of Lexington, plants covering spring-fed farm pond, 11 Jun 1987, Haragan 1528 (Univ. Ky. Coll. Agric. Herbarium: KNK): Harrison Co., off KY 356, ca. 2 miles e of Cynthiana, farm pond, 3 Jul 1996, Meade 2284 (MOKY, Morehead State University Herbarium); Kenton Co., nr. Independence, farm pond, 23 May 1994, Meade 1511 (MOKY, Morehead State University Herbarium). We acknowledge aid from Kathy Stewart and Nancy Hopkins. LITERATURE CITED. (1) Beal, E.O., and TW. Thier- et. 1986. Aquatic and wetland plants of Kentucky. Kentucky Nature Preserves Comm., Sci. Tech. Ser., 5:1-314. (2) Webb, D.H., and E.W. Chester. 1989. Distribution of the homed pondweed, Zannichellia palustris L. (Zannichelli- aceae), in Tennessee and Kentucky. Trans. Kentucky Acad. Sci., 50:158—160.—Les Meade and Brian Binion, De- partment of Biological and Environmental Sciences, More- head State University, Morehead, KY 40351; Peggy Mea- sel and Brenda Hamm, T.H.E. Engineers Inc., Lexington, KY 40509; and Patricia Dalton Haragan, Herbarium, University of Louisville, Louisville, KY 40294. Trans. Ky. Acad. Sci. 58(2):101. 1997. List of Reviewers for Volume 58 of Transactions of the Kentucky Academy of Science John R. Baird Richard I. Barnhisel David M. Brandenburg Jerry Carpenter Ronald R. Cicerello Michael A. Flannery Michael A. Floyd Larry A. Giesmann Vernon Hicks Ellis L. Laudermilk James O. Luken Max E. Medley Robert F.C. Naczi Debra K. Pearce 101 Thomas C. Rambo Arthur Raymond James Sickel David Sleper Jerry W. Warner Stephen White Steven Wilkinson Trans. Ky. Acad. Sci. 58(2):103-110. 1997. INDEX TO VOLUME 58 Compiled by Varley Wiedeman A-chain gene transcription, 42-43 Aesculus glabra var. glabra, 15 Abstracts, 37-54 Accipitridae, 54 Acer saccharinum, 87, 88 rubrum, 89 Acidic mines, oil effects on nodula- tion, 80-84 Actinonaias ligamentina, 56, 57 spectorosa, 57 AD-like pathology, in normal aging, 4l Adiantum capillus-veneris, 74 ADKINS, MARCI, 43 Aesculus glabra var. glabra, 13 African-American health and diet status, 39 rural southern elderly, 39 Agricultural sciences, abstracts, 37- 38 Agrypnia vestita, 69, 71, 72 Alasmidonta atropurpurea, 97 Alkylaton of DNA, 40 ALLEN, PETER, 99 Allium cepa, 60 Alpheus heterochaelis, snapping ac- tion of, 52-53 Alzheimer’s PS-1 mutation, 40 Ambrosia, 17 Ameiurus nebulosus, 96 American ipecac, 12, 16 American lotus, 13, 16 American pennyroyal, 12, 16 American swallow-tailed kite, 98 Ammocrypta clara, 97, 98 pellucida, 98 vivax, 98 Andropogon arenaceum, 15 avenaceus, 15 nutans, 15 spp., 21 Anguispira rugoderma, 97 Animals of Kentucky, rare and extir- pated, 96-99 Annonaceae, 37 Antimonite, sensing system for, 42 Aplodinotus grunniens, 96 Appalachian false-foxglove, 14, 16 Aquila chrysaetos, 97 Ardea alba, 98 ARMSTRONG, JAMES S., 54 Arnoglossum muehlenbergii, 12, 15 Arrow-wood, Missouri, 98 Arundinaria gigantea, 12, 15 Asarum canadense, 18 virginicum, 14, 15, 18 Ashcamp cave beetle, 98 Asian clam, 96 Asimina triloba, 13, 15, 37 Asperula odorata, 18 Asteraceae, 17, 38 Astroglial toxicity opiate-mediated, 46 TNF-alpha neuroprotection in, 46 Astronomy for non-science majors, 48 use of computers for, 48 Astronotus ocellatus, gentamicin-in- duced toxicity in, 49, 50 Aureolaria flava,18 laevigata, 14, 15, 18 patula, 74, 97 BACKUS, JASON, 23 Bahalana ‘eracei, 53 effects of cannibalism on, 53 from san Salvador Island, Baha- mas, 53 population structure of, 53 Bald eagles densities, 54 distributions, 54 impacts of dam construction on, 54 Baldcypress, 85, 88 Baptisia australis, 12, 15 cerulea, 12, 15 coerulea, 12, 15 tinctoria, 75 Barren oats, 15, 16, BARRETT, BILL, 40 Bartonia paniculata var. paniculata, 75 virginica, 75 Bastard pennyroyal, 12, 16 BEATTIE, RUTH E., 51 Beetle cave, 98 concealed cave, 98 scholarly cave, 98 schoolhouse cave, 98 stone-dwelling cave, 98 surprising cave, 98 Berchemia scandens, 97 Beta vulgaris, 65 Bidens discoidea, 87 Bieberbach groups, classifying free, 29—32. BILES, DANIEL C., 92 BINION, BRIAN, 100 103 Biology, student attitudes toward, 51 Birds, 97, 98 BITNER, RONALD D., 53 Bitter milkwort, 14, 16 Bivalva, 55-59 in the Cumberland River, 55-59 Black darter, 4 Black wort, 15, 16 BLACK, RICHARD, 42 BLAIR, PETER, 23 BLANK, SARAH M., 46 BLOOM, C. TOM, 74 Blue catfish, 96 predation on zebra mussel, 96 Blue false-indigo, 12, 16 Blue penny-royal, 14, 16 Blue wild indigo, 12, 16 Blue-curls, 12, 16 Blue-hearts, 15, 16 Bluestems, 21 Body weight in rats effect of dietary energy restric- tion, 49 effect of ovariectomy on, 49 Boehmeria cylindrica, 87 BONDADA, SUBBARAO, 43 BOOZE, ROSE-MARIE, 41 Botanical regions in Kentucky, 20- 22 Constantine Rafinesque’s 1819 description of, 20-22 Botany and microbology abstracts, 38-39 Botany of Kentucky, 12 BRACKNEY, CHRISTOPHER, 41 Bradyrhizobium, 80-84 Breast cancer, human, 46-47 Briar, sand, 17 BRICKING, KEITH D., 42 Brown bullhead, 96 BROWN, C., 49 BRYANT, WILLIAM S., 20, 85 Buchnera americana, 15 Buck-eye, prickly, 13, 17 Buckeye, Ohio, 13, 17 Buffalo, smallmouth, 96 Bull chub, 96 Bullhead, brown, 96 Button wood, 13, 16 Buttonbush, 87 Buttonwood, 16 Cacalia, 17 atriplicifolia, 17 muehlenbergii, 12, 15, 17 reniformis, 12, 15 Caddisflies, of the Kentucky River, 67-73 Calcium excretion, urinary, 49 104 Transactions of the Kentucky Academy of Science 58(2) Calcium homeostasis, 40 neurotaxicity, 4] metabolism, in rats, 50 Calopogon tuberosus, 75 tuberosus var. tuberosus, 74 Cambarus ornatus, 97 CAMMERS-GOODWIN, ARTHUR, 40 Campostoma anomalum, 6 Cane, 12, 16 giant, 12, 16 Cannibalism, 53 effects on population structure, 53 Capraria multifida, 13, 15 Carex atlantica ssp. capillacea, 98 crawei, 75 stricta var. stricta, 75 CARINI, MICHAEL, 48 CARLSON, SONIA, 44 Carp, common, 96 CARPENTER, JERRY H., 53 CARRICO, LYNN, 41 Carya illinoinensis, 89 Casmerodius albus, 98 Castanea dentata, 75 Catalpa, 18, 20 bignonioides, 18 cordifolia, 18 northern, 13, 17 speciosa, 13, 15, 18, 20 tree, 13, 16 Catalpium cordata, 13, 15 cordatum, 18 cordifolium, 13, 15, 18 Catfish, blue, 96 predation on zebra mussel, 96 Catostomus commersoni, 6 Caucasian health and diet status, 39 rural southern elderly, 39 Cave beetle, 98 ashcamp, 98 concealed, 98 scholarly, 98 schoolhouse, 98 stone-dwelling, 98 surprising, 98 CCDs, photometry with, 48-49 Cedar, red, 14, 17 Celestial seasons, 47 Celithemis verna, 98 Cenchrus longispinus, 76 Center for automated space science, 48 Cephalanthus occidentalis, 87 Cephalogonimidae, 23-28 Cephalogonimus vesicaudus, 23-28 Ceraclea ancylus, 69, 71 cancellata, 69, 71 flava, 69, 71 punctata, 69, 71 tarsipunctata, 69, 71 transversa, 69, 72 Ceratopsyche sparna, 69, 70 Chenopodium ambrosioidaes var. anthelminti- cum, 12, 15 anthelminthicum, 12, 15 Cheumatopsyche campyla, 69—71, 73 minuscula, 69, 70 oxa, 69, 70, 73 pasella, 69, 70 Chimarra obscura, 69, 70, 72 Chromosome 17 deletions, in ovar- ian tumors, 41—42 Chub, bull, 96 CICERELLO, RONALD R., 55 Cichlidae, 49, 50 Clam, Asian, 96 Clammy-weed, 13, 16 CLEMENTS, SANDRA, 47-49 Cliff plumb, 16 CLINE, GARY R., 80 Clover, nimble, 15 Common carp, 96 Common laurel, 14 Common names, of vascular plants, 9-19 Community college graduates, 50- 51 COMPTON, KEVIN, 40 Computerized education, a student survey, 51-52 Computers, use for astronomy, 48 CONARD, NYREE, 38 Concealed cave beetle, 98 Coneflower eastern, 14 orange, 14, 17 purple, 14, 17, 21 Conobea, 17 COON, ALEXANDER, 41 Copper iris, 16 Corbicula fluminea, 56, 96 Cotton tree, 13 Cottontails, eastern, 1-3 cover, 1-3 habitat, 1-3 in Kentucky, 1-3 Cottonwood, 13, 16, 20 Cow mint, 13, 16 CRAWFORD, MELINDA L., 52 Crested dwarf iris, 14, 16 Crested flag, 16 Crested iris, 14, 16 Crested tris, 14, 16 Crustaceans, 97 Cryopreservation of seeds, of Ohio wetland plants, 38 CUILARAN, FONSIE, 48 CULLER, RYAN, 47, 48 Cultured panicle segments, 60-66 Cumberlandia monodonta, 58 Currant, wild, 14, 17 CURTSINGER, W. STEWART, 49 Cuscuta sp., 87 Cuterebra abdominalis, 35 buccata, 35 fontinella, 35 sp., 35-36 Cyclonaias tuberculata, 55, 57-58 Cyperus croceus, 76 difformis, 76 Cyprinus carpio, 96 Cyrnellus fraternus, 69, 70 Cytochrome P450 2E1, 45-46 Cytokine production, 42 Cytokines, and neonatal immune re- sponses, 43-44 Dalea candida, 15 purpurea var. purpurea, 15 Ddarter black, 4 saffron, 4 duskytail, 98 Ddarters, snubnose, 4-8 separation of spawning habit, 4-8 spawning habit in, 4-8 DAUNERT, SYLVIA, 42 DEARTING, TERRY L., 53 DECKARD, MONICA, 47, 48 DEDRICK, ERIC, 45 Deerberry, 14, 16, 21 DELISLE, BRIAN, 43 DELONG, JASON E., 42 DEMOSS, D.L., 50 DERTING, TERRY L., 51, 54 Dietary energy restriction, effects of on rats, 49 Diptera, 35-36 Distinguished scientist award, 33 Double-ringed pennant, 95 Drum, freshwater, 96 Duskytail darter, 98 Dwarf iris, crested, 16 Eastern coneflower, 14, 16 Echinacea purpurea, 14, 15, 21 Echinostoma trivolvis, 23-28 Echinostomatidae, 23-28 EGAN, RITA, 42 EIZENGA, GEORGIA C., 60 Elanoides forficatus forficatus, 98 Electrophysiological studies, of the SCN8a_ voltage-gated _ sodi- um channel, 43 Elephant'’s foot, 12, 16 leafy, 12, 16 Elephantopus caber, 12, 15, 17 carolinianus, 12, 15, 17 ELLINGTON, JEFF, 23 ELLIOTT, CHARLES L., 1 Ellipsaria lineolata, 55, 57 Elliptio crassidens, 56, 57 dilatata, 56, 57 Elodea nuttallii, 97 EMBRY, KENNETH, 39 Emden-Fowler equation, polyno- mial conservation laws of, 40 Endangered Ohio wetland plants, cryoperservation of seeds of, 38 Epilobium brachycarpum in Kentucky, 99 ciliatum var. ciliatum, 76 paniculatum, 99 Epioblasma brevidens, 58, 97 capsaeformis, 58, 97 EPSCoR opportunities for space-re- lated science, technology, 48 Esox masquinongy, 46 Etheostoma barrenense, 5 blennioides, 6 caeruleum, 5, 6 cinereum, 97 clarum, 98 crossopterum, 6 davisoni, 5 duryi, 4, 5, 7 flabellare, 6 flavum, 4—S flavum, spawning in, 4—8 pellucidum, 98 percnurum, 98 rafinesquei, 5 simoterum, 4-8 simoterum, spawning in, 4—S smithi, 6 tippecanoe, 97 vivax, 98 zonale, 5 Ethics development and content, 51 science, 51 world view and values, 51 Eupatorium celestinum, 15 coelestinum, 13, 15 rugosum, 13, 15 urticefolium, 13, 15 Extirpated animals of Kentucky, 96- 99 Extirpated plants of Kentucky, 96— 99 Extragalactic radio jets, 47 Fabaceae, 80-84 FALBO-KENKEL, MARIA, 47 False pennyroyal, 14, 16 False-foxglove, Appalachian, 14, 16 False-indigo, blue, 16 Fawnsfoot, 96 Fescue, tall, 60-66 Index to Volume 57 characterization of for DNA changes, 60-66 characterization of for isozyme changes, 60-66 characterization of for meiotic changes, 60-66 Festuca arundinacea, 60 FIALKOW, JARED, 52 Field investigations, 51 educational benefits, 51 misconceptions, 51 problems, 51 student perspectives of, 51 FINNESETH, C.L.H., 37 Fire-pink, 15, 16 Fishes, 97, 98 Flag, 14, 16 crested, 16 Forestiera acuminata, 87, 88 Fraxinus pennsylvanica, 87, 88 profunda, 87 Free Bieberbach groups, classifying, 29-32 FREEMAN, JAMES, 44 Freshwater drum, 96 Fringed-orchid orange, 14, 17 yellow, 14, 17 FRYE, TRICIA L., 96 Gaultheria procumbens, 14, 21 GELDERMAN, MICHAEL CAR- INI, 47 GELDERMAN, RICHARD, 47, 48 Genes involved in splicing, 44 Gentamicin-binding protein(s), 49— 51 in ear of cichlid fish, 49-50 Gentian stiff, 15, 17 yellow, 15, 17 Gentiana alba, 98 amarelloides, 15, 18 amerellvides, 15 flavida, 98 quinquefolia var. occidentalis, 18 quinquefolia var. quinquefolia, 15, 18 Geography abstracts, 39 Gerardia glabrata, 14, 15 Giant cane, 12, 16 GIESMANN, LARRY A., 38 GILBERT, NIKOLE L., 43 Gillenia stipulacea, 12, 15 stipulata, 12, 15 GILLIS, PETER P., 40 GIULIANO, WILLIAM M., 1 Glossosomatidae, 69, 70 Golden-rod, stiff, 14, 17 Goldenrod, stiff, 17 105 Graduates, of community colleges, 50-51 GRANESE, JACQUELINE M., 52 Grass, Indian, 15, 16, 21 Gravitational lensing simulation, 47 Great Indian-plantain, 12, 16 Green ash, 87 GUILARAN, ILDEFONSO, 47 GUO, QING, 40 HACKNEY, KAREN, 47-49 HACKNEY, RICHARD, 47-49 Hairy ruellia, 15, 16 Halesia tetrapera, 97, 98 Haliaeetus leucocephalus, 54 densities, 54 distributions, 54 impacts of dam construction on, 54 HAMPTON, C. TONY R., 53 HAMPTON, D., 49 HARAGAN, PATRICIA DALTON, 100 HARP, CINDY, 44 Hatching, in the muskellunge, 46 stage dependent induced, 46 HAUSER, KURT F., 46 Health and diet status, differences in, 39 Health sciences abstracts, 39-40 Heart-leaf, 14, 16 Hedeoma pulegioides, 12, 15 Helicodiscus notiusspecus, 98 punctatellus, 98 Heliconia, 53 latispatha imbricata, 53 extrafloral nectaries, 53 Heliconiaceae, 53 Heliotrope, 13, 16 Indian, 13, 16 Heliotropium indicum, 13, 15, 17 Helisoma trivolvis, 23-28 Heracleum lanatum, 98 maximum, 98 HEROD, JEFFREY J., 96 Hesperis pinnatifida, 13, 15 Hexastylis, 18 virginica, 14, 15 Hippocampal subregions, 41 HOGSTRAND, CHRISTER, 40 Horned-pondweed, 99-100 Horse nettle, 16 HOUP, RONALD E., 67 Houstonia, 18 angustifolia, 18 fruticosa, 13, 15 nigricans var. nigricans, 13, 15 tenuifolia, 18 Houstonia, narrow-leaved, 13, 17 Howe sedge, 98 HT 1080 human fibrosarcoma can- cer cells, 42 106 Transactions of the Kentucky Academy of Science 58(2) HUANG, HONGWEN, 37 Human breast cancer, mRNA in, 46-A7 Human myiasis in Kentucky, 35-36 Human pancreatic carcinoma, 44— 45 Hydropsyche betteni, 69 frisoni, 69, 70 hetteni, 70 orris, 67, 69, 70 simulans, 69, 70 Hydropsychidae, 69, 70 Hydroptila ajax, 69, 70 angusta, 69, 70 armata, 69, 71 consimilis, 69, 71 hamata, 69, 71 perdita, 69, 71 waubesiana, 69, 7 Hydroptilidae, 69, 7 Hymenocallis caroliniana, 13, 15, 18, 20 liriosme, 18 occidentalis, 18 Hypericum perforatum, 99 walteri, 87 oe Ictaluridae, 96 predation on zebra mussel, 96 Ictalurus furcatus, 96 predation on zebra mussel, 96 Ictiobus bubalus, 96 Immune responses, neonatal, 43-44 Immune tissues, innervation of, 44 role of nerve growth factor, 44 Indian grass, 15, 16, 21 Indian heliotrope, 13, 16 Indian physic, 12, 16 Indian-plantain, great, 16 Indigo, blue wild, 16 Induced hatching, stage dependent, 46 Innervation of immune tissues, 44 role of nerve growth factor, 44 Insecta of the Kentucky River, 67— 13 Insects, 97, 98 lodanthus pinnatifidus, 13, 15 Ipecac, American, 12, 16 Iris cristata, 14, 15 crocea, 13, 15 fulva, 13, 16 Iris copper, 16 crested, 14, 16 crested dwarf, 14, 16 Iron weed, 16 Ironoquia punctatissima, 69, 71-73 Ironweed, 12 tall, 17 Isanthus brachiatus, 14, 16 caeruleus, 16 ceruleus, 14, 16 coeruleus, 14 Isopod, marine cave, 53 JACKSON, JANN, 46 Jackson Purchase Region, of Ken- tucky, 85-91 Jeffersonia binata, 13, 16 cinata, 13, 16 diphylla, 16 JENNINGS, JOHNNY, 48 JONES, S.E., 40 Juncus validus, 74, 76 Juniperus virginiana, 14, 16, 21 JUST, JOHN J., 46 KAETZEL, DAVID, 42 Kalmia latifolia, 14, 21 KENTUCKY STATE NATURE PRESERVES COMMISSION, 99 Kentucky vascular plants, 74-79 Kentucky's Jackson Purchase Re- gion, 85-91 Kidney weed, 12, 16 Kite American swallow-tailed, 98 swallow-tailed, 98 KRUPER, JILL H., 51, 53 L-type calcium channel density, 41 Lampsilis cardium, 56, 57 fasciola, 57 ovata, 56, 57 Large-flowered leafcup, 14, 16 Lasmigona costata, 57 LAUDERMILK, ELLIS L., 55 Laurel common, 14 mountain, 2] LAYNE, DESMOND R., 37 Leafcup large-flowered, 14, 16 yellow-flowered, 14, 17 Leafy elephant’s-foot, 12, 16 LEATHERS, MICHELLE JO, 38 Lechea, 18 minor, 14, 16 LEE, C.J., 39, 49 Lepomis cyanellus, 6 megalotis, 6 microlophus, 96 Leptoceridae, 69, 71 Leptodea fragilis, 58 Lespedeza cuneata, 80-84 sericea, 80-84 Leucospora multifida, 13, 16, 17 LEWIS, BRIAN M.., 40, 47 LIBBY, GARY W., 74 Ligumiarecta, 57 Lilium michiganese, 77 superbum, 76 Lily, 13, 16 red, 13, 17 swamp, 17 Limnephilidae, 69, 72 Linum usitatissimum, 77 List of reviewers, 101 Little brown jugs, 14, 17 Littorinidae, 52 LIU, BIN, 42 LOHRE, JOE, 47 Lolium perenne, 65 Lonicera dioica var. orientalis, 97 Lotus, 18 American, 13, 16 yellow, 13, 17 LOU, X., 50 LUO, JIANYUAN, 44 Lupatorium calutinum, 13, 16 Luxilus chrysocephalus, 6 Lycopus rubellus, 87 Lythrurus ardens, 6 Macrhybopsis gelida, 97 meeki, 97 MALPHRUS, BENJAMIN K., 40, 47 Maple, red, 89 MARLETTE, MARTHA, 39 Mathematics abstracts, 40 MATTSON, MARK P., 40, 46 McGRUDER, CHARLES, 48 McGRUDER, MICHAEL CARI- NI, 47 McNEIL, RAYMOND C., 48 MEADE, LES, 100 MEARS, RANDY L., 74 MEASEL, PEGGY, 100 Medicago sativa, 65 Medionidus conradicus, 57 MELLON, ISABEL, 43 MERCER, JANET, 42 Metal impact in the Robinson For- est, using metalliothionein, 40— 4] Metalliothionein, assessment of metal impact, 40-41 Miagia arupedinaria, 12 arupendinaria, 16 Micropterus punctulatus, 6 Microsatellite instability and TGF-b Type II receptor gene muta- tion, 44-45 Miegia arundinaria, 12, 16 Migraine headaches, effects of weather systems on, 39 Milkwort bitter, 14, 16 racemed, 14, 17 MIMS, S.D., 38, 50 Mink, 56 Mint, cow, 16 Mismatch repair, 43 Mississippi alluvial plain, swamp for- ests of, 85-91 Missouri arrow-wood, 98 Mist-flower, 13, 17 Molecular and cell biology abstracts, 40-47 Morehead radio telescope, 47-48 optimization of operating param- eters, 40 surface geometry of, 40 MORRISON, JOHN R., 51 Mountain laurel, 21 Mountain tea, 14, 21 MPTP-treated C57B1/6 mice, 45 mRNA in human breast cancer, translational regulation of, 46— 47 Muhlenbergia cuspidata, 77 Muskrats, 56 Mussel, zebra, 96 in the Ohio River, 96 Mussels, 97 Mutagenesis of a conserved phenylalanine, 45— 46 site directed, 45-46 Mutations, in the RNA sorting sys- tem, 45 Mycena leaiana, fine structure of bacidiospores, 38 Mystacides sepulchralis, 69, 71, 72 NACZI, ROBERT F.C., 39 Nanostoma, 4 Narrow-leaved houstonia, 13, 17 NASA Kentucky space grant consor- tium, 48 Nectopsyche candida, 69, 72 exquisita, 69, 72 pavida, 69, 72 Nelumbium pentapetalum, 13, 16 Nelumbo, 18 lutea, 13, 16 Neotyphodium coenophialum, 60 Nephrotoxicity, gentamicin-induced, Nettle horse, 16 white, 13, 17 Neureclipsis crepuscularis, 69, 71, 1 Nicotinein MPTP-treated C57B1/6 mice, 45 Nicotine, neuroprotective effects of, 45 Nimble clover, 15 Nimble weed, 14 Index to Volume 57 Nimbleweed, 17 Nitrogen-fixing bacteria, 80-84 nm23 proteins, in the WR-082-01 cell line, 42-43 NOBLE, JOHN, 47, 48 Nocomis raneyi, 96 Nodulation, effects of acidic mine soil, 80-84 Northern catalpa, 13, 17 NORTHINGTON, ALAN, 45 Notes, 35-36, 96-100 Notropis albizonatus, 98 sp., 98 NOVOTNY, NATHAN, 45 Nucleotide excision repair mutants, 43 Nymphaea sp., 77 Nyssa aquatica, 85, 87, 88, 90 = Oaecetis avara, 72 cinerascens, 72 inconspicua, 72 persimilis, 72 Oats, barren, 16 Obliquaria reflexa, 55, 57 Ochrotrichia aergerfasciella, 69, 71 cristata, 69, 71 tarsalis, 69, 71 xena, 69, 71 Oecetis avara, 69, 71 cinerascens, 69 inconspicua, 69, 73 persimilis, 69, 73 Oestridae, 35-36 Ohio buckeye, 13, 17 Ohio wall flower, 13, 17 Onagraceae, 99 ONDERS, R., 38 Optical monitoring, of quasars, 48 Orange coneflower, 14, 17 Orange fringed-orchid, 14, 17 Orchis ciliaris, 14, 16 Orconectes inermis, 97 Oryza sativa, 65 OTTE, ELIZABETH, 44 Otter, 56 Outstanding teacher award college/university, 33 secondary school, 33-34 Ovariectomy, effects of on rats, 49 Ovariectomy surgery effects of on rats, 49 effects on urinary calcium excre- tion, 49 Overcup oak, 87 Owsley Fork Reservoir, 23-28 Oxyethira pallida, 69, 71 Pa-paw tree, 13 Paddlefish, 50 body weight, 50 107 muscle lipid content, 50 stability of meat stored in crushed ice, 50 using flood control reservoirs for, 38 Palezone shiner, 98 PAN, BIN-TAO, 49 Pancratium liriosme, 13, 16, 18 liviosone, 13, 16 Pancreatic carcinoma, 44—45 Panicle segments, cultured, 60-66 PARRISH, MARK, 44 PAULY, JAMES R., 45 Pavia muricata, 13, 16 Pawpaw, 13, 17 allozyme variation in, 37 geographic differentiation in, 37 influence of shade on, 37-38 morphological development, 37— 38 root-zone modification, effect on, 37-38 seed germination, 37-38 seedling emergence, 37-38 Pawpaw tree, 17 Pea-clover, yellow, 14, 17 Pecan, 89 PENCE, VALERIE C., 38 Pencil flower, 14, 17 Pennant, double-ringed, 98 Penny royal, 12 Penny-aoyal, 12, 17 Penny-royal, 17 blue, 14, 16 Pennyroyal American, 12, 16 bastard, 12, 16 false, 14, 16 Percidae, 4—8 Percina caprodes, 6 evides, 97 Peromyscus leucopus, 35 Petalostemon candidum, 16, 21 moncandidum, 15 purpureum, 15, 16, 21 Petalvitemon candidum, 15, 16 Phenylalaninein cytochrome P450 2E1, 45-46 PHILLIPS, TIMOTHY D., 60 Philopotamidae, 69, 72 Phlox glaberrima, 14, 16 Phlox, smooth, 14, 17 Photometry with CCDs, 48-49 optimizing signal extraction, 45— 49 Photomorphogenic control, in dan- delion, 38 of flower scape elongation, 38 Phyrganeidae, 69, 72 Physic, Indian, 12, 16 108 Transactions of the Kentucky Academy of Science 58(2) Physics and astronomy abstracts, 47-49 Physiology and biochemistry ab- stracts, 49-52 PIERETTI, MAURA, 41 Pimephales notatus, 6 Pin weed, 14, 17 Pine, pitch, 14, 17, 21 Pink, 14, 17 scarlet, 15, 17 Pinus rigida, 14, 16, 21 Pinweed, small, 14, 17 Pitch pine, 14, 17, 21 Plagiorchis micracanthos, 26 Planera aquatica, 87, SS Planning a trip, 92-95 Planorbidae, 23-28 Plants, 98 Plants of Kentucky, 96-99 rare and extirpated, 96-99 Platanthera ciliaris, 14, 16 Platanus occidentalis, 13, 16, 20, 88 Platelet-derived growth factor, 42— 43 Plum, wild, 13, 17 Polanina graveolens, 13, 16 Polanisia dodecandra ssp. dodecandra, 13, 16 graveolens, 13, 16 Polycentropodidae, 69, 72 Polycentropus cinereus, 69, 72 confusus, 69, 70, 72 Polygala paucifolia, 74, 77 polygama, 14, 16 senega, 39 senega var. latifolia, 39 senega var. senega, 39 Polymnia uvedalia, 14, 16 Polynomial conservation laws, of the generalized Emden-Fowlser- equation, 40 Polyodon spathula, 38, 50 Polysaccharides, responses to, 43-— 44 Populus angulata, 13, 16 deltoides, 13, 20, 88 deltoides var. deltoides, 16 Porcelia tribuba, 13, 16 triloba, 13, 16 PORTER, TODD, 45 Porteranthus stipulatus, 12, 16 PORTERFIELD, JEAN C,, 4 Potamilus alatus, 56-58 ohiensis, 58 Potamyia flava, 69, 70 Potentilla recta, 99 Prairie vole, 53-54 female preferences, 53-54 male odors in, 53-54 Prairie-clover purple, 15, 17 white, 15, 17 Prairie-clovers, 21 Prairie-dock, 15, 17, 21 Prickly bog sedge, 98 Prickly buck-eye, 13, 17 PRINGLE, JAMES S., 9 PRINS, RUDOLPH, 52 Proterometra macrostoma, 27 Protoptila maculata, 69-71 Prunus munsoniana, 13, 16, 18 pendula, 13, 16, 18 Pseudanophthalmus abditus, 98 catoryctos, 97 hypolithos, 98 inexpectatus, 98 pholeter, 97 scholasticus, 98 Ptilostomis ocellifera, 69-72 semifaciata, 69-72 Ptychobranchus fasciolaris, 57 subtentum, 55-58 Punctate coil, 98 Purple coneflower, 14, 17, 21 Purple prairie-clover, 15, 17 Purple rocket, 13, 17 Purple sun-flower, 14, 17 Pycnopsyche lepida, 69, 72, 73 Pyganodon grandis, 58 Pyrimidine biosynthesis enzymes localization of, 44 with fusion proteins, 44 Quadrula pustulosa, 56, 57 Quasars optical monitoring of, 48 very high red shift, 48 Quercus lyrata, 87, 88 Raccoons, 56 Racemed milkwort, 14, 17 Radio frequency astrophysics, 47-48 Radio jets, extragalactic, 47 Radio telescope, 47-48 Rafinesque’s 1819 description of bo- tanical regions, 20-22 Ragweed, sand, 13, 17 RAMANATHAN, SRIDHAR, 42 RAMBO, THOMAS C., 53 Rare animals of Kentucky, 96-99 Rare plants of Kentucky, 96-99 Rats effect of dietary energy restric- tion, 49 effect of ovariectomy on, 49 effects on urinary calcium excre- tion, 49 RAWLS, JOHN M., 44, 45 Red cedar, 14, 17 Red lily, 13, 17 Red maple, 89 Red-eared sunfish, 96 Redcedar, 21 REID, JAMES, 42 Retinioc acid effect on stromelysin, 42 effect on TIMP, 42 Rhyacophila lobifera, 69, 71-73 Rhyacophilidae, 69, 72 Rhynchospora macrostachya_ var. macrostachya, 74, 77 RICHMOND, RAYMOND E., 42 RIGDON, STEVEN M., 49 RNA sorting system, in spermiogen- esis, 45 Roach, 96 ROBBINS, MARK C., 38 ROBINSON, DAVID LOWELL, 38 ROBINSON, MARK P.,, 92 Rock weed, 13, 17 ROSEN, RON, 23 Rough bell, 15, 17 Rough wort, 14, 17 Rubiaceae, 18 RUCH, DONALD G., 38 Rudbeckia fulgida, 14, 16 fulgida var. fulgida, 14 purpurea, 14, 16 Ruellia caroliniensis ssp. ciliosa, 15, 16 oblongifolia, 15, 16 Ruellia, hairy, 15, 16 Rutilis rutilis, 96 RYMOND, BRIAN, 44 Saffron darter, 4 Salix nigra, 87, 88 San Salvador Island, Bahamas, Tec- tarius muricatus on, 52 Sand briar, 13, 17 Sand ragweed, 13, 17 SAPP, TAMARA, 46 SATIN, JONATHAN, 43 Saururus cernuus, 87 Scarlet pink, 15, 17 Scented sun flower, 14, 17 SCHEFF, STEVE, 41 Schistosoma japonicum, 27 mansoni, 27 Schoenoplectus mucronatus, 74, 77 purshianus, 78 Scholarly cave beetle, 98 Schoolhouse cave beetle, 98 SCHUSTER, GUENTER A., 67 Science ethics development and content, 51 for biology and chemistry majors, Sl world view and values, 51 Scirpus mucronatus, 74, 77, 78 purshianus, 78 Sciuridae, 54 Sciurus carolinensis, 54 in woods of western Kentucky, 54 leaf-nest composition, 54 nest-site selection, 54 SCN8a voltage-gated sodium chan- nel, electrophysiological studies of, 43 SCOTT, ROGER, 47-49 Sedge Howe, 98 prickly bog, 98 SEGEBARTH, BRAD, 46 Seneca snakeroot, taxonomic status of, 39 Sensing system, for antimonite, 42 Sericea lespedeza, 50-84 SHIBER, J.G., 50, 51 Shiner, palezone, 98 Shrimp alpheid, snapping action of, 52 Silene catesbaei, 16 catesbei, 15, 16 catesbri, 15, 16 virginica, 15, 16 Silphium terebinthaceum, 16 terebinthinaceum, 16, 21 therebinthaceum, 15 Silver maple, 87 SILVERNAIL, ANTHONY F., 80 SINEX, MATTHEW, 42 SISKEL, JAMES B., 96 Skeletal compartmentalization, in rats, 50 Sky weed, 13 Sky-weed, 17 Small pinweed, 14, 17 Smallmouth buffalo, 96 SMITH, CHRISTOPHER L., 40, 47 Smooth phlox, 14, 17 Snailnatural infection of trematodes, 23-28 Snails, 97, 98 Snakeroot, white, 13, 17 Sodium channel, SCN8a_voltage- gated, 43 Solanum carolinianum, 13, 16 caroliniense, 13, 16 phureja, 65 SOLE, JEFFERY D., | Solidago rigida, 14, 16 Sorghastrum nutans, 15, 16, 21 Sorghum bicolor, 65 Southern rural elderly dietary knowledge, 39 health status, 39 intakes of, 39 Index to Volume 57 SPARKS, LARRY, 41 Spearmiogenesis, RNA sorting sys- tem in, 45 Spider-lily, 13, 17 Spiderlily, 20 Splicing, genes involved in, 44 SPRAKER, JOHN S., 92 STANFORD, VINCENT EVIN, 54 Stedeoma pulegioides, 16 pulegivides, 12 Stiff gentian, 15, 17 Stiff golden-rod, 14, 17 Stiff goldenrod, 17 Stinking weed, 13, 17 Stone-dwelling cave beetle, 98 Strain-induced electrophilic cyclic rings, 40 STRAUSS, A.M., 40 Streptococcus pheumoniae, 43 STUCKEY, RONALD L., 9 Student attitudes, toward study of biology, 51 Stylosanthes biflora, 14, 16 elatior, 14, 16 Stylvianthes elatior, 14, 16 Sun flower scented, 14, 17 purple, 14, 17 Sunfish, red-eared, 96 Surprising cave beetle, 98 Swallow-tailed kite, 98 American, 98 Swamp forests composition of, 85-91 of Mississippi alluvial plain, 85-91 structure of, 85-91 Swamp lily, 13, 17 SWIDERSKI, CAROL, 44 Sy-weed, 13, 17 Sycamore, 13, 17 Sylvilagus floridanus, 1-3 cover, 1-3 habitat, 1-3 in Kentucky, 1-3 Synandra grandiflora, 13, 16 hispidula, 13, 16, 18 Tall fescue, 60-66 characterization of for DNA changes, 60-66 characterization of for isozyme changes, 60-66 characterization of for meiotic changes, 60-66 Tall ironweed, 12, 17 Taraxacum officinale, 38 Tavin weed, 13, 17 Taxodium distichum, 85, 87, 88, 90 Taxonomic status, of varieties of Seneca snakeroot, 39 Tea, mountain, 14, 21 109 Tectarius muricatus on San Salvador Island, Bahamas, 52 size and density of, 52 Teleostei, 4—S TEMPLETON, SUSAN, 39 TENNANT, RAYMOND F., 29 TGF-b Type II receptor gene mu- tations, 44-45 THIERET, JOHN W., 99 THOMAS, ELIZABETH M.., 40 TNF-alpha neuroprotectionin opi- ate-mediated astroglial toxicity, 46 Traditional education, a student sur- vey, 51-52 TRAPASSO, L. MICHAEL, 39 TRAUTH, AMY E., 39 Trematodes daily emergence, 23-28 effect of light on, 23-28 Trepocarpus aethusae, 97 Triaenodes ignitus, 69, 72 tardus, 69, 72 Tricholomataceae, 38 Trichoptera, of the Kentucky River, 67-73 Trip planning, 92-95 Triplasis purpurea, 97 Tris, crested, 14, 16 Triticum aestivum, 65 Tritogonia verrucosa, 56, 57 Truncilla donaciformis, 96 truncata, 55, 57 Tupelo, water, 85, 88 Turnsole, 13, 17 Turpentine weed, 15, 17 Twin leaf, 13 Twin weed, 17 Twinleaf, 17 Ulmus americana, 88 Unionid, freshwater, 55-59 in the Cumberland River, 55-59 Unionidae, 55-59 in the Cumberland River, 55-59 URSO, JAN, 45 Utterbackia imbecillis, 58 Vaccinium album, 14, 16 stamineum, 14, 16, 21 VARAJAN, RALPH CHEL, 43 Vascular plant discoveries, from Kentucky, 74-79 Vascular plants common names of, 9-19 in Kentucky, 9-19 reported by C.S. Rafinesque, 9- 19 Vernonia gigantea ssp. gigantea, 12, 16 praealta, 16 110 Transactions of the Kentucky Academy of Science 58(2) prealta, 12, 16 Viburnum lentago, 97 molle, 98 Villosa iris, 57 lienosa, 57 trabalis, 56-58 Vole, prairie, 53-54 female preferences, 53-54 male odors in, 53-54 VUJANOVIC, B.D., 40 Wall flower, Ohio, 13, 17 WALLACE, DAVID, 41 WANG, C., 38, 49, 50 Water tupelo, 85, 88 WATSON, CASEY, 47, 48 Weather systems, effects on mi- graine headaches, 39 WEAVER, DANA R., 44 White nettle, 13, 17 White prairie-clover, 15, 17 White snakeroot, 13, 17 WHITE, T.L., 50 WILBUR, FRANK H., 51 Wild currant, 14, 17 Wild indigo, blue, 16 Wild plum, 13, 17 WOODWARD, JEROLD W,, 42 Worm weed, 12, 17 Wormseed, 12, 17 WR-082-01 cell line, 42-43 WRIGHT, G.L., 50 YAN, HONG Y., 49, 50, 52 Yellow fringed-orchid, 14, 17 Yellow gentian, 15, 17 Yellow lotus, 13, 17 Yellow pea-clover, 14, 17 Yellow wort, 14, 17 Yellow-bunch, 14, 17 Yellow-flowered leafcup, 14, 17 Zannichellia palustris, 99-100 Zannichelliaceae, 99—100 Zebra mussel, 96 in the Ohio River, 96 ZHANG, Y., 49 ZIMMER, STEPHEN, 46 Zoology and entomology abstracts, 52-54 ‘ie Wy 0 ily nda aa dhe sua HECKMAN [= BINDERY INC. ll OCT 99 n...1 7. pit..@ N. MANCHESTER. 4] wu oc < ig 4] aT r 4 fe) = =) = — 2) = Zz < Zz fe) a” xr E = o” 3 9088 01304 3369 pemeninne se cretyees AMOR AES aE ae vn wgeienies napa ren ah peers bere Bm PAPA Tams Nery Pet Aare Mt =" a BRERA MEYRIN AS ee ARK Syren Banner Oi poresy stil ee rer etee eases Coaalbetes FOX PLEAS tid ony 82 OP EN etry ‘ a Mim ( Aaa e ee LOY z SN et ar lal aba a RCE Tae DEM ae eg PAVE a ty SA suPneyite SY E AEM UU HIM RS SN sans ROASAUE RAY ABR ALE PANU Mea Wwe hieae Samar AIN US oS Wasi Pees tire amine UWS At ot, s Um ata PAIR WBRATE SOWA NN Me 8) MAMAS > aneiew Uren, SPAR Veta errno PNPRE EE VERRY PRN VTA WAP Moma RSE LA Bist ply vinegar “Sve SUN ets Vege iA PUP arAAAP We Fae : Lay Adar MONI EYE. ere aiaetr tates st ry APNeneten fataen Pat SA A tieminte Baimy Wy an Cra laate is Uryyauses WEEE MBP RALEA Don ay Aub drt ge ee pate a y ° BLRSBAP AER rN ARENA EAP SENN NPL by ARSE UA LPVS VIA BEEN or MENA SARWAN ER EAT OPA DRT EMPE OAT EAP BOAT 3 Ue APAPAE CABS ENDER TPs APYANE Eee tr UA eAD EE SAE trar teaar SAA APE ADAG GSP AP 4 Caras LPabas ASTER NP STAT VERE NVA Pees hie4 ay AP OASAG VATE PLEALA PN UNLESS HELAA TH pet rune atts VEMPAT UP UR UR AU) af OL Stet Cage SU Vest UEDA sare leis as Vkae PAN CR VEDP ED ag ASE AE Nt \ AAG A ach tient VA Meats SEAR vim) 17 sare RES Ruta ic tl NERV ESEAT Ey barns ben ait op ob ALS PAT EAE PVE eAR SA athe PAP PURE NASD DE LR 1 DAPTEN RET ENE SENPSIN OSPR AES Pe pase PURE ENS RUA PST Dupe ene gare ceayacwmas a2 WPM ES O55 ANE OT ened NA RG ER DEE AG NUP VTA Ly NAPPA REMAOAES OU ppc ecties SRESPATSDSESEAD OPT ETERD ENERO ENE A x SeAaRE SREP RESP NEARER Mee ee arty TRAPP MIAU APA t re eats PREMIER TS NSRP IR Borers eget Pyke peep ker ae: setts: FINE NE MOR NO Oger Narn bees utesanenrrenets ore ary pt tet TA LST Maes whem ee teereny! asaya. Der nyo wre en winrar ta au We een os OER A pecniremgmy vin ate “s +6 M SEAMEN AEN SPAN Bs Mey Re MRAP Se AeA RE Ne NPenarSRAENO tenes TAPNE iP SEN ENLALN ES PS ES NE DSO ES LSPS PATS, renee MIRE EMED IN TN EREY, MPRA EASE PS SAM ERPS IN SAPARD EN PP IAS NS A Aust Ne Parra NPE A Prop UAE SUE Ny re ers ie SORE SPAIRND NEE Ry SPUN SRY NU eh tap PM Urs Dar nensmeriereom pat} : POs ny ey li aE A Se DRE NETL ND iit odd ad PARA PEED EAS PUP APT EEN PYERENKA TALS ONIBEL SNPS ERE RD A SPN LAD Ale NEND Of EAE CA CR RCL URU STE STL Trot SEU ADAa gE 8, rsa ea vane ennngs 5 AONE INESP ERY PapAr aad TEA DAN or DSPE EEN EREAN RAPS EDEN ESTE AN Rh teay MT NIAH DREN LEA NESE Up Ni sy spans MEENA E REREAD EP EF, ABR OD SA NMUEND ED to wg? Leese. PAN Pale beset & AP DP NDB AEA rier ter SOREN P ERD BLEND Agape SEA SAT MS METS LEB RRA MERE be per el RENAL EE AVEND REE j Fe Iau) Der Pete ESE wpahe 1 DRAB ALUN ERASE MIND a IPN re Read Yee te eee DESIR rae VE MEAP PRIA Pe AE TEA SORE TERY Pura TENE RIVE MAA Vierrnysy ENP RE SES Soars : arate re BARRE A Br DECRERAPRE ries SA Sa oy e eh baa ee one re! PAE sath ese 2 r= AG ih MSL pee hg apy MRRP HS APE TNT MAPIRMEM Pmaneyemian po je got ge SABRENT RPI FR Bh RRA Pho Mir Nev Ot erate aw ait OD dT ee x nied earong tye essed enreeecsittn nye eee ard Siidde tite ores APRON A SAP PRB ere ety: iets Un Keay xp Dies tira iar ade en eae Eras Aone Sete ants " 4 VA Ssk's SHON Yt cea AEE chide eg rd eae 2259 Sens enna raid att eit Pac toe PY DUES OE EEY *Blmenemar rege $' take bees 158 Se NbRis CL Ree ORY ree ev One Sy puree rayne) PAR Harte a LUNAP MR ER IDA Abreartens