Historic, Archive Document Do not assume content reflects current scientific knowledge, policies, or practices. & >»««£ >vcuir<^ ■V \^nn5«st 2»iiiw*£j S*&nnjflf£ 5*22?, >jm*<£ ^tutr^ 5*»®»S S^niOrf^, ytiMCUi;^ >'Ciar^ ^inirt^V. ^>75 'ffllS* W** 5«®iss gowns' ^unri^L fyj fltt'' &»»«£ £t»lM% ^cuir^ ^vd sr^ $#1M£ Hr $$mHr y5MCiiir«j£5 as mi'v yWWir .Am** v^\cmr^ iSJtt «l* ,-wcuir^ V>*'culty2 S»«n3V^j hs q^ SjMMf#/ SI© it4ff ^LX aVCU (tuff yw»»«S fetyniw^V ii©mh*a ^VJwwiJ^s ^vcutr^ OFFICIAL PUBLICATION OF THE VIRGINIA ACADEMY OF SCIENCE Virginia Journal of Science Volume 34, Number 1 Spring 1983 / J Dynamic Analysis of Vein Calcite in an Area of Non-coaxial Deformation S. Andrew Orrell and Charles M. Onasch Department of Geology University of Maryland College Park, MD 20742 Abstract - Stress orientations determined from twinned vein calcite in three generations of shear and extension fractures, now veins, were compared to those predicted on the basis of vein orientation and movement sense. The determined and predicted stress orientations compare closely for extension veins, but not for shear veins. Dilation of exten¬ sion fractures, filling with calcite, and twinning of the calcite all occur under the same uniform stress that produced the extension fractures. Twinning of calcite in shear fractures is caused by stresses unrelated to those present during shear fracturing. Subsequent stresses have only minor effects on calcite in extension veins, but may greatly alter calcite fabrics in shear veins. Introduction An understanding of the relationship between stress and the resulting structural features is one of the basic goals of structural geology. To this end, several techniques for the determination of stress orientations and magnitudes have been developed, largely through the experimental deformation of rocks and minerals. Of these, the most widely used is the analysis of twin lamellae in calcite. Calcite has been used successfully to determine stress orientations in a variety of geologic settings ranging from slightly deformed sedimentary rocks to highly strained metamorphic rocks (for review, see Friedman and Sowers, 1969 ; Carter and Raleigh, 1969). Studies utilizing primary and recrystal 1 ized calcite in sedi¬ mentary rocks or recrystallized calcite in metamorphic rocks are limited to areas of relatively simple deformational history. In areas of non¬ coaxial deformation, the calcite will have been subjected to several different stress systems. Resolution of these superimposed stresses from a single generation of calcite generally is not possible. Calcite in syn-tectonic veins has been largely overlooked for dynamic analysis. Veins offer great promise for dynamic analysis because they are nearly ubiquitous in deformed rocks. Most originate as syn- tectonic fractures that are subsequently dilated and filled with minerals such as calcite. In an area of non-coaxial deformation, it is common for several generations of veins to develop, each reflecting a different stage in the stress history. Twinning of calcite in each vein should record the incremental stress orientations present during the formation of that vein. By analyzing a number of veins formed during different increments of deformation, reconstruction of the superimposed stress systems should be possible. This study investigates this possibility by comparing stress orientations derived from twinned vein calcite to those predicted inde¬ pendently on the basis of vein orientation and movement sense in rocks that have undergone non-coaxial deformation. 4 THE VIRGINIA JOURNAL OF SCIENCE Sample Descriptions Samples for this study are from the Ordovician Lincolnshire Formation exposed in the Shenandoah Valley near Strasburg, Virginia (Fig.l). Structural ly , the area is on the western limb of the Massanutten synclino- rium, a doubly-plunging asymmetric fold. Bedding in this location is relatively planar and dips uniformly to the southeast at approximately 40 degrees. Here, the Lincolnshire Formation is a fine-grained biomicrite with abundant chert nodules. Several generations of calcite-filled shear and extension veins are present. Two sets of extension veins, both occurring in an en echelon fashion were found. Most individual extension veins are planar, but some are slightly sigmoidal. Shear veins can be differentiated from extension veins by their offset of fossils, chert nodules, and other veins. Veins vary in thickness from 0.3 to 1.5 cm with extension veins as a group being somewhat thicker. Sparry cal cite fills both vein types and increases in grain size toward the center of the vein. Fibrous cal cite, typical of many syn-tectonic veins (Ramsay, 1980) was not observed in these samples. Some of the cal cite in the oldest shear veins is brecciated and cemented with a second generation of cal cite. No evidence of a replacement origin was observed and we believe that all veins originated by the filling of dilated fractures. Based on cross-cutting relations, three generations of veins were identified. These are shown schematically in Figure 2. The first, or oldest, generation consists of extension veins only. Both shear and extension veins formed during the second generation. Contemporaniety of the shear and extension veins in this generation is established by mutual offset and examples where one type passes uninterupted into the other (see Fig. 2). The third, or youngest, generation consists of shear veins only. Outcrop relief was sufficient to allow accurate measurement of vein orientations. Relative movement directions were determined from slickenside orientations on the vein walls and from offset of older veins or other features. Mutually perpendicular thin sections were prepared from oriented samples collected from each vein type in each of the three generations. The orientation of cal cite c-axes and twin lamellae were measured with a universal stage, plotted on an equal area stereonet, and rotated back to true geographic coordinates. Fig. 1. lOO km Index map of Virginia showing Shenandoah Valley (ruled pattern) and study area near Strasburg. DYNAMIC ANALYSIS OF VEIN CALCITE 5 Fig. 2. Schematic diagram showing relative ages and orientations of veins. Diagram would correspond approximately to a vertical east-west cross section with west to the left. Stress Analysis Stress orientations were determined for each set of veins from the dynamic analysis of the twinned vein calcite. In order to test the hypothesis that these orientations represent incremental stress orienta¬ tions related to the formation of the vein, an independent and accepted technique for the determination of the stresses is required. For this purpose, stresses were predicted on the basis of the vein orientation and movement sense. With these data and assuming that all veins origi¬ nated as fractures, stress orientations can be predicted from brittle yield theory. Predi cted Stress Orientations According to brittle yield theory, shear and extension fractures form at certain angles to the principal stress axes (Price, 1966; Anderson, 1951). In a general compressive stress field (aHXjpXJo), such as would be expected in an area undergoing folding, extension fractures form normal to the least principal stress, a~. Shear fractures form in a conjugate set that intersects along the intermediate stress axis, c^. The maximum principal stress, a, , bisects the acute angle between the shear fractures which is typically about 60°. Where only one of the conjugate shear fractures is observed, as is the case for the third generation of shear fractures, a, will lie at an angle of 30° to the fracture in a plane defined by the pole to the fracture and the slip direction (slickensides) in a direction consistent with the relative movement sense of the fracture, cu will be normal to a-, in this same plane and will be normal to both a-jand a^. Using these relationships, principal stress orientations were predicted for each generation of veins (Fig. 3). For the first generation, consisting of extension veins only, a unique determination of a, and is not possible. Both will lie in the great circle shown in Figure 3a. The relative orientations of shear and extension veins in the second generation is consistent with brittle yield theory, further supporting thei r synchronous development. 6 THE VIRGINIA JOURNAL OF SCIENCE Dynami c Analysis of Vei n Cal ci te Stress orientations were determined from twinned cal cite in each vein according to the method of Turner (1953). Twinning on the e plane (01 T2) in cal cite is known to be a mechanical ly- induced phenomenon. For any grain, the compression axis (a,) most favorable for twinning is oriented at an angle of 45° to the twin plane and 71° to the c-axis, in the plane containing the pole to the twin plane and the c-axis. The tension axis (a~) is normal to the compression axis in the same plane. Compression ana tension axes are determined for a number of grains yielding a statis¬ tical orientation of a, and a~, respectively. The point of the highest concentration on a contoured diagram is taken as the orientation of the axis for that diagram. It should be noted that this method actually determines the axes of strain (shortening and extension) and that the axes of stress are derived by assuming a coaxial relationship between stress and strain. Rocks deformed in the laboratory typically exhibit a coaxial stress-strain relationship (Friedman, 1963). For Turner's method to be valid, the cal cite grains must be randomly oriented in order that all twin lattice directions have equal chance to twin. The lack of any preferred orientation in the c-axis fabrics of the veins studied indicates that this condition is met. Figure 4 shows the compression and tension axis diagrams for each vein. Several observations can be made from these plots. (1) The compression and tension axis fabrics of older veins are less well defined than those of younger veins. (2) For each vein generation, tension axis fabrics are generally better defined than compression axis fabrics. (3) For the second generation, the compression and tension axis fabrics of shear veins are less well defined than those of extension veins. (4) For the second generation, compression and tension axes determined from the extension vein do not agree with those from the shear vein. By comparing the determined to the predicted stress orientations (Fig. 5), it is apparent that stresses determined from twinned cal cite in extension veins agree well with those predicted from vein orientation (Fig. 5a & b). In contrast, stress orientations determined from cal cite in shear veins show no correlation to those predicted from vein orientation or to those determined from extension veins of the same age (Fig. 5c & d). Discussion The close correlation between stress orientations determined from cal cite in extension veins and the lack of correlation in shear veins can be explained by differences in the evolution of the two vein types. Extension veins formed from extension fractures that were imme¬ diately dilated and filled with cal cite. The cal cite was twinned before the stresses responsible for the fracture had changed orientation. Hence, the stresses recorded by the twinning of the cal cite are the same as those responsible for the fracture. Subsequent stress systems had relatively minor effects on the cal cite in extension veins, dispersing the compression and tension axis fabrics somewhat. Rotation of existing twin lamellae and formation of a limited number of new twins are responsible for the broader compression and tension axis maxima in the older extension veins. The original fabric, however, remains largely intact. Apparently, once the cal cite is twinned during the initial increment of stress, twinning by additional stress increments becomes less likely. In shear veins, the lack of agreement between stress orienta¬ tions determined from cal cite twins and those predicted from vein orien¬ tation is due to a more complex dilation and filling history. At the time of shear fracturing, the normal stress acting across the fracture was sufficient to prevent dilation and filling. Only when the stresses had changed orientation with respect to the fracture, could the fracture dilate and fill with cal cite. Twinning of the cal cite occurred at a later time by stresses unrelated to those responsible for the initial GENERATION 2 GENERATION 3 GENERATION Predicted stress orientations for the three generations of veins. Vein orientations plotted as great circles. Slickensides on shear veins indicate relative movement directions. In the first generation (a), O] and o2 both lie in the plane of the vein. Fig. 4. Contoured compression and tension axis diagrams for the three generations of veins. Contours at 1.5, 6, and 9% per 1% area. Veins plotted as great circles. The number of lamellae sets measured are 68, 62, 72, and 61 for diagrams a, b, c, and d, respectively. 8 THE VIRGINIA JOURNAL OF SCIENCE 1** GENERATION 2nd GENERATION 3rd GENERATION 1st GENERATION 2"° GENERATION 3,u GENERATION extension extension shear shear DYNAMIC ANALYSIS OF VEIN CALCITE 9 Comparison of stress orientations determined from calcite twin lamellae to those predicted from vein orientation and movement sense for the three generations of veins. Note agreement between determined and predicted stresses in extension veins (a and b) and lack of agreement in shear veins (c and d). 10 THE VIRGINIA JOURNAL OF SCIENCE shear fracture. Therefore, the stress orientations derived from cal cite in shear veins bear no relationship to those predicted from vein orien¬ tation or to those determined from extension veins of the same fracture generation. The cal cite in shear veins records a later dilation and filling event. It is possible that the orientation of the shear veins is not in accord with brittle yield theory. Preexisting anisotropies could have caused the shear fractures to form in orientations different from those predicted by brittle yield theory. However, the homogeneous nature of the beds sampled would argue against this. It also appears that shear veins were affected more than ex¬ tension veins by subsequent stress systems. Dispersal of compression and tension axes occurred not only by rotation of twin lamellae and new twin¬ ning, but also by renewed shearing and brecciation of the original vein filling. Twinning of the cal cite that cements the vein breccia fragments occurred at an even later time; hence, it is unrelated not only to stresses responsible for the shear fractures, but also to those associated with twinning of the initial generation of vein cal cite. Shearing and brec¬ ciation were not observed in extension veins. This could be a result of their shape and extent. Long, continuous shear veins may have been more susceptible to subsequent shearing than the short, discontinuous, en echelon extension veins. In summary, this study indicates that dynamic analysis of twinned cal cite in extension veins may be used to determine the incre¬ mental stress oreintations present at the time of vein formation. Dila¬ tion of the extension fractures, filling with calcite, and twinning of this calcite occur in a geologically short period of time, before the stresses responsible for the fractures change orientation. Because di¬ lation and filling of shear fractures are not immediate, twinned calcite in shear veins does not record stress orientations present during forma¬ tion of the shear fractures. Dilation, filling, and twinning of the calcite in shear veins occur after the stresses responsible for the fractures have changed orientation. Subsequent stress systems may alter, but do not destroy, the original compression and tension axis fabrics of extension veins. Each vein records the stress orientations present during its formation. In an area where several generations of extension veins are present, a detailed record of the stress history is preserved. Interpretation of this history requires only that the relative age of each vein be known. Literature Cited Anderson, E. M. 1951. The Dynamics of Faulting. Oliver & Boyd, Edinburg. 206 pp. Carter, N. L. , and Raleigh, C. B. 1969. Principal stress directions from plastic flow in crystals. Geol . Soc. America Bull. 80: 1231-1264. Friedman, M. 1963. Petrofabric analysis of experimental ly deformed calcite-cemented sandstones. Jour. Geol. 71: 12-37. Friedman, M. , and Sowers, G. M. 1969. Petrofabri cs : A critical review. Canadian Jour. Earth Sci . 7: 477-497. Price, N. J. 1966. Fault and Joint Development in Brittle and Semi- brittle Rock. Pergamon Press, New York, 176 pp. Ramsay, J. G. 1980. The crack-seal mechanism of rock deformation. Nature 284: 135-139. Turner, F. J. 1953. Nature and dynamic interpretation of deformation lamellae in calcite of three marbles. Am. Jour. Sci. 251: 276-298. Virginia Journal of Science Volume 34, Number 1 Spring 1983 /2.P Microorganisms Isolated From Jthinocyllus conicus1, A Weevil Introduced into Virginia for Thistle Control. P. I . I)«»d; and L. T Department of Entomology Virginia Polytechnic Institute and State University Blacksburg, Virginia 24061 ABSTRACT Flower heads of musk thistle, Carduus thoermeri Weinmann, infested with Rhino cyll us conicus Froelich were collected to study the prevalence of potentially pathogenic microorganisms in R. conicus in Virginia. In 1978 and 1979, weevil-infested flower heads from three field sites located in Pulaski, Frederick and Warren Counties were caged in the laboratory until adult weevils emerged. Dead larvae and adults of R. conicus remaining in the thistle heads were screened for microorganisms associated with each insect stage. Several species of bacteria were isolated from the larvae, the most common being Bacillus cereus Fr. & Fr. and 13. megatherium DeBary. A fungus Beauveria bassiana (Bals.) was one of the few pathogens isolated from adult weevils. Rate of infection was low in both years. Hemolymph and gut tissues of healthy weevils, as well as host plant tissue did not harbor any pathogenic organisms. INTRODUCTION Rhinocyllus conicus Froelich was imported from Europe into the U.S.A. and released in 1969 for the control of musk thistle (Carduus thoermeri Weinmann) and plumeless thistle (Carduus acanthoides L.). The weevil is presently well-established in Virginia, and in several other states (Surles et al. 1974, Rees 1977, Goeden and Ricker 1977, Puttier et al. 1978). Impact of predation and parasitism on R. conicus has been reported (Dowd and Kok 1981, 1982), but no documentation of the incidence of pathogenic microorganisms in the various populations of the weevil in the U.S.A. is available. Because we have occasionally encountered evidence of diseased weevils during field surveys, the prevalence of potential pathogens in R. conicus was investigated in 1978 and 1979 as part of our study of mortality factors affecting this introduced weevil. METHODS AND MATERIALS One hundred musk thistle flower heads containing large numbers of developing larvae of R. conicus were collected from each of 3 field sites Coleoptera: Curculionidae ► 'Present Address: Dept, of Entomology, LSU, Baton Rouge, LA. 12 THE VIRGINIA JOURNAL OF SCIENCE Table 1. Pathogenic microorganisms isolated from dead larvae and adult Rhinocyllus conicus in Virginia. Microorganism Type1 Host life No. of stage sites detected 2 % R. conicus infested 1978 1979 Bacillus megatherium B larva 2 12.5 0 Bacillus cereus B larva 2 20.3 0 adult 1 3.1 0 Serratia sp. B larva 1 6.3 0 Streptococcus sp . B larva 1 3.1 0 Micrococcus sp . B larva 1 3.1 0 adult 2 6.3 0 Beauveria bassiana F adult 2 0 5.9 Fusarium F larva 2 4.7 0 = Bacteria, F = Fungus 2 Based on specimens that were dead and dissected from thistle heads: 96 larvae and 96 adults in 1978, and 60 larvae and 60 adults in 1979 located in Pulaski, Frederick and Warren counties, VA during mid-June, 1978. The thistle heads were individually held in 0.9£ clear plastic containers in the laboratory and dissected in August after weevil adults emerged. Dead specimens totaling 192 (32 larvae and 32 adults/site) that remained within the heads were randomly selected for screening of associated microorganisms. Whole specimens were surface sterilized by the technique of Poinar and Thomas (1978), ground in distilled water, and inoculated onto plates of nutrient agar. The resulting bacterial colonies were identified by colony morphology, Gram stains, endospore stains, catalase, Voges-Proskauer , and cytochrome oxidase tests. Fungal colonies were slide mounted in Guegen's solution and examined. Representative colonies of microorganisms were sent to the diagnostic laboratories at the University of California, Berkeley, and to the U.S.D.A. Agriculture Research Center, Beltsville, MD, for confirmation. In 1979, another 60 specimens of dead larvae obtained from musk thistle flower heads from the same field sites were screened for associated microorganisms. Over 500 adults field collected in June, 1978, and held in equal numbers (100/treatment) at 5 different temperatures (15°, 20°, 25°, 30°, 35°C) in the laboratory, and 500 adults overwintered in field cages were examined for signs of fungi at time of death. Samples of 60 specimens were sent for screening of pathogenic microorganisms. In addition, hemolymph and gut tissue were removed from over 50 healthy adult weevils dissected from thistle heads, plated, and examined. Resulting colonies from a random sample of 12 weevils were sent to diagnostic laboratories for identification. Sections of bud tissue from 12 thistles randomly selected from over 50 buds examined, were treated in the same manner to determine the presence of pathogenic microorganisms in host plant tissue. Only 12 samples of healthy weevils and bud tissue were sent for identification because our initial examinations showed a lack of potential pathogens in them. RESULTS AND DISCUSSION No pathogenic microorganisms were isolated from the gut or hemolymph of healthy weevils collected from thistle flower heads or from the thistle bud tissue. Five bacteria and 2 fungi were found in dead larvae and adults. Of the potential pathogens isolated from the larvae in 1978 (Table 1), two bacteria, Bacillus cereus Fr. & Fr. and _B. megatherium DeBary, were encountered most frequently. Both species, however, are also known to occur widely in soil and other material (Buchanan and Gibbons 1974) and their invasion may have been secondary. Two bacteria isolated from the adult weevils dissected from thistle flower heads in 1978 were Micrococcus sp. and Bacillus cereus. A fungus, Beauveria bassiana (Bals.), was isolated from 5.9% of the adults held in the laboratory from 1978 into 1979 and MICROORGANISMS FROM AN INTRODUCED WEEVIL 13 from adults overwintered in field cages. No pathogenic organisms were isolated from larvae in 1979. The absence of pathogens in thistle bud tissue, and the hemolymph and gut tissues of healthy weevils, suggests that the isolated microorganisms are invaders and are not a normal component of healthy weevils or thistle tissue. Some of the isolated pathogens (Serratia sp . , Streptococcus sp., Beauveria bassiana and Bacillus cereus) have been previously reported infecting other Curculionidae (Weiser 1969, Buchanan and Gibbons 1974, Cantwell 1974, Thomas and Poinar 1973, and Poinar and Thomas 1978) . Due to the widespread occurrence of several of the pathogens and their known ability to invade as secondary agents, their importance in relation to R. conicus appears to be more as secondary rather than primary invaders. Although adults do not seem to be frequently attacked, about one-third of the samples of dead larvae in 1978 contained microorganisms which were potentially pathogenic. This suggests that disease microorganisms may play an important role in contributing to mortality in the larval stage, even though the pathogens are not always the primary cause of death. The absence of pathogens in larvae in 1979, however, indicates that incidence of disease in,R. conicus varies greatly between years, and is probably influenced by the prevailing predisposing conditions for infection. The low infection rate and absence of disease epidemics in the various field populations of R. conicus which have been established for over ten years confirm that pathogenic microorganisms are not major mortality factors of the weevil. ACKNOWLEDGEMENTS Research was supported in part by USDA/SEA Cooperative Grant #12-14- 1001-1204 and EPA Grant #R8059 3 1-02-0 . We thank Dr. J. L. Vaughn of the Plant Protection Institute Insect Pathology Laboratory, USDA/ESA, Beltsville, Md., for identification of the microorganisms. LITERATURE CITED Buchanan, R. E. and N. E. Gibbons (eds.). 1974. Bergey’s manual of determinative bacteriology. Williams and Wilkins Co., Baltimore, MD. 1246 pp. Cantwell, G. E. (ed.). 1974. Insect diseases. Vol. I. Marcel Dekker, Inc. New York. 300 pp. Dowd, P. F. and L. T. Kok. 1981. Predators of Rhinocyllus conicus (Coleoptera: Curculionidae) in Virginia. Environ. Entomol. 10: 136-8. Dowd, P. F. and L. T. Kok. 1982. Parasitism of Rhinocyllus conicus (Col.: Curculionidae) in Virginia. Environ. Entomol. 11: 71-7. Goeden, R. D. and D. W. Ricker. 1977. Establishment of Rhinocyllus conicus on milk thistle in southern California. Weed Sci. 25: 288-92. Poinar, G. 0., Jr. and G. M. Thomas. 1978. Diagnostic manual for the identification of insect pathogens. Plenum Press. New York. 218 pp. Puttier, B., S. H. Long, and E. J. Peters. 1978. Establishment in Missouri of Rhinocyllus conicus for the biological control of musk thistle (Carduus nutans) . Weed Sci. 26: 188-90. Rees, N. E. 1977. Impact of Rhinocyllus conicus on thistles in southwestern Montana. Environ. Entomol. 6: 839-42. Surles , W. W. , L. T. Kok, and R. L. Pienkowski. 1974. Rhinocyllus conicus establishment for biocontrol of thistles in Virginia. Weed Sci. 22: 1-3. Thomas, G. M. and G. 0. Poinar, Jr. 1973. Report of diagnoses of diseased insects, 1962-1972. Hilgardia 42: 261-352. Weiser, J. 1969. An atlas of insect diseases. Irish University Press. Shannon, Ireland. 282 pp. Virginia Journal of Science Volume 34, Number 1 Spring 1983 An Evaluation of Small Rodents in Four Dismal Swamp Plant Communities F. Elizabeth Breidling, Frank P. Day, Jr., and Robert K. Rose Department of Biological Sciences Old Dominion University Norfolk, Virginia 23508 ABSTRACT Diversity and density of small rodents in the Dismal Swamp are believed to be low. Reasons for this may be excessive predation, heavy interspecific pressure from large rodents, lack of suitable habitat, low food availability or flooding. Rodent populations were evaluated using live-traps and pitfall traps in four different Dismal Swamp plant communities. Habitat was compared on the basis of phytomass studies previously reported. Flood levels were recorded during 1 i ve - 1 ra ppi ng sessions. Mast from trees was collected in modified mast collectors, and fed to P e r o m^s c u s leuco^us in the laboratory. Only two small rodent species were captured: Ochrotora^s and Pe r o m^_s c u s leucopus. The low density of these animals is believed to be limited by lack of food combined with unsuitable habitat. Flooding appears to have a great effect on small rodent diversity by preventing the establishment of species that are not at least mediocre swimmers and somewhat arboreal. INTRODUCTION Rodent population studies conducted in the Dismal Swamp have historically yielded low densities and diversity (Handley 1 979)- There are at least five factors which may limit the rarity and density of small rodents in a swamp habitat. These are excessive predation, pressure from interspecific competition, sparse undergrowth (or cover), extensive flooding, and insufficient food. Population studies of Dismal Swamp predators are practically non-existent, but it is generally believed that the predator numbers are low. This also appears to be the case with larger rodents, such as rats and squirrels, so these animals are unlikely to impose a severe interspecific competitive pressure on the small rodents . The importance of dense undergrowth to small rodents has been demonstrated by several workers (e.g., Miller and Getz 1977, Goodpaster and Hoffmeister 1954, M’Closkey and Lajoie 1975). However, a paucity of comparisons exists between rodent species diversity in mesic versus hydric habitats. A few rodents can survive well in hydric habitats, but the number of different species which can survive is obviously restricted by the necessity of being scansorial and natatorial. The role of food in SMALL RODENTS IN THE DISMAL SWAMP 15 controlling the abundance of many vertebrates has been well documented in the literature; Lack (1954), working with birds, was the first researcher to emphasize this regulatory role of food . The objectives of our study were to (1) qualitatively and quantitatively evaluate the small rodent populations of four vegetatively different Dismal Swamp stands, and (2) investigate three factors which have the potential to influence these populations, that is: flooding, undergrowth density, and food availability . Description of the Study Area The Great Dismal Swamp National Wildlife Refuge was created in 1974 when the Union Camp Corporation deeded a large tract of land to the Nature Conservancy, who in turn donated it to the U.S. Department of Interior (Berkely and Berkely 1976). Parcels of land continue to be added to the Refuge; at present, the Refuge comprises some 40,000 ha. However, the Dismal Swamp proper extends beyond the current Refuge boundaries and has been estimated by Oaks and Whitehead (1979) to be about 85,000 ha. Most of this area is predominantly a peat swamp; peat formation is believed to have commenced about 11,000 BP (Whitehead and Oaks 1979) . The biota of the Dismal Swamp has been considerably disturbed by man, starting in the mid-l8th Century, when canals were dug to promote drainage and to transport lumber. Swamp lowlands were later drained for agricultural purposes, considerably reducing the area of the Swamp (Stewart 1979) and altering the water table. The reduction of virgin swamp forest is directly attributable to these human disturbances. Most plant communities consist of second or third growth forest, and include several different serai stages (Levy and Walker 1979). The area is consequently losing many of its swamp characteristics. Four plant stands, chosen as representative examples of the heterogeneity of the Swamp, have had their vegetation structure quantified by Dabel and Day (1977). These stands were the study areas for this research on small rodents, and the undergrowth density data of Dabel and Day's study were used in determining the effect of habitat upon small rodent populations. The composition of the four forest stands is summarized in Table 1 (from Dabel and Day 1977). The mixed hardwood stand was in a mesic area with mineral soils (Histic Humaquepts). Its shrub layer was moderately dense, as was its herbaceous layer (due more to the presence of tall canebrakes than to a dense low ground cover). The maple-gum stand had a shrub layer which was less dense than that of the mixed hardwood stand, and an extremely sparse herbaceous layer. This stand previously contained cypress (Taxodium dist ichum ) trees, but these have been unable to reproduce because of competition from maple trees. Similarly, poor germinating conditions prevail in the cypress stand. The lack of cypress seedlings here may prevent replacement of this species; the hardwoods, whose seeds are germinating successfully, are increasing in importance. The cedar stand had a dense shrub stratum made possible by canopy openings caused by the death of the Atlantic white cedar trees, and red maple and blackgum were replacing the cedar trees. Cedar's failure to reproduce may be due in part to the exclusion of fire by man. The mixed hardwood stand is inundated sporadically and unevenly during the late winter and early spring. During this same period the other stands are usually flooded too. Cypress is the most frequently flooded, maple-gum is next, and cedar, though least flooded, occasionally is covered to 30 cm depths. 16 THE VIRGINIA JOURNAL OF SCIENCE Table 1. Dominant tree species in four Dismal Swamp plant communities (Dabel and Day 1977). Data represent relative total biomass, expressed as a percentage. Mixed Maple- Cedar Cypress Hardwood gum Tree Species Quercus laurifolia 32.8 Quer cus alba 17.1 Liquidambar stryaciflua 13*1 Nyssa sylvatica 11.8 Acer rubrum 9.5 Nyssa aquatica Taxodium distichum Chamaecyparis thyoides 16.6 30.9 14.5 32.9 19.5 18.0 46.7 46.8 50.1 Field Procedures MATERIALS AND METHODS In order to evaluate the food supply, mast collections were begun during the spring of 1979, using 15 litter collectors that had been modified to make them bird- and m a m m a 1 - pr oo f . These collectors consisted of wire mesh baskets 10 cm deep, with a basal area of 0.25 m2, placed on wooden legs 50 cm tall. The modifications included the addition of wire mesh strips (7.6 cm wide) around the lower periphery of the basket, preventing rodents from climbing into the basket, and a square of chicken wire (with openings of 2.5 cm) placed on top of the mesh basket, to prevent deer and birds from eating the mast. The collectors, previously used for other studies, had been placed along two parallel lines, at random distances from each other. Bimonthly mast collections were made from September 1 979 to July 1 980 on all four stands. Population studies of the small rodents were conducted in May and October 1979 and September 1980. The 1979 studies used live traps, the latter used pitfall traps. Two lines, 125 m long and 40 m apart, were measured in each stand, and 100 Sherman live traps (10 x 10 x 25 cm) were placed along these lines at 5 m intervals, with two traps at each station. The mixture of peanut butter and oats used as bait in May decomposed rapidly, so sunflower seeds were used in October. The traps were checked daily, and captured mice were marked with an indelible pen, for estimating population density. Water depth was measured at each stand during both live- trapping sessions in 1979. For the pitfall trapping, grids were set up, using four lines 20 m apart, with seven traps 10 m apart on each line. Holes were dug at these grid points and cylindrical 10 tin cans were inserted into the holes. Water was placed in these cans to a depth of 10 cm. Traps were checked every fourth day for 10 days. Laboratory Procedures Mast collected at the four stands was oven-dried at 70° C for 48 hours, and then separated by species and weighed. In order to determine which species of mast are edible by SMALL RODENTS IN THE DISMAL SWAMP 17 small rodents, "cafeteria" tests were performed in the spring and summer of 1980, using wild-caught P e r o m^s£u s leucopus , white¬ footed mice, which had been acclimated to the laboratory, then weighed prior to each use. One specimen, used as a control, was maintained on sunflower seeds. Mast species collected in the Dismal Swamp were fed to two mice. After two days on a diet consisting exclusively of a tree seed, the mice were re-weighed and, if no inter pre table results were obtained, the experiment was continued for another two days, or was repeated later. The mast being tested was also weighed at the beginning and end of the experiment; thus, the food consumed could be quantified, as well as the weight losses or gains of the animals. If part of an edible fruit was rejected by the mice, or caused weight loss, it was considered indigestible. In order to determine relative digestibility of each edible fruit, the digestible portion of several samples of each species was recovered and weighed. In order to determine whether collection time and site differences had a significant effect on mast production, one-way analysis of variance (ANOVA) tests were run on the biomass of the total mast collected. A priori tests were also run, testing for significant differences between groups. The mast biomass data from the mixed hardwood stand were compared with the other stands, as the mixed hardwood stand is a mesic area and the other stands are periodically inundated. In addition, the mast data from the cedar stand were compared with the remaining plant communities, because the cedar stand is the one stand with a true peat soil. In order to look at summer and winter production, July and September biomass estimates were averaged to obtain a summer value for comparison to the mean of January and March; in addition, January’s production was compared to July’s. Autumn and spring production were compared using November and May values. RESULTS Small Rodents Only in the cedar community were both species of rodents, white-footed mice and golden mice (Ochro tomys nuttalli ) , trapped (Table 2). All four communities had low numbers of rodents. Once marked, none was recaptured, so no population estimates could be calculated. The highest yield of white-footed mouse specimens was in the maple-gum community. Production of mast The mixed hardwood community produced the greatest variety, but the smallest quantity of mast (Table 3). Six species produced between 10 and 25 percent of the annual total. More acorns were found here than at the other communities but they were concentrated in a few baskets. The maple-gum stand produced the largest amount of mast for the year. The primary contributor, with 81.5 percent, was tupelo gum. Few acorns were found. The distribution of mast species was rather uneven, as indicated by the standard errors. Maple had the most even distribution, whereas oak mast was only found at one basket . Five different species produced mast in the cypress stand, with tupelo gum (68.3$) the most important. Blackgum, maple, and sweetgum fruits were found in small quantities. Cypress cones were unique to this stand, but were only found in a few collectors. The cypress stand’s mast was the least evenly distributed of all the communities. The cedar stand produced the second most abundant mast, but 18 THE VIRGINIA JOURNAL OF SCIENCE this represented only three species. The majority was produced by Atlantic white cedar, but fruits were found in greater quantity at the cedar site than at other sites. Distribution of cedar cones and maple samaras was noticeably even, more so than mast at the other communities. Table 4 shows the amount of mast, by species, collected at bimonthly intervals during the year. The small amount o'f mast produced in late spring and summer is notable (Table 4); red maple contributed most then. Red maples produced fruits only Trom March through May. Tupelo, however, was found mainly in the winter months. Acorns were found at the mixed hardwood stand in every collection period, with most in January, and least in July. However, acorns were never found in abundance. Table 2. Flood data and results of live trapping in May and October 1979. Each plot was trapped for 400 trap nights during each season. Mixed Maple- Cypress Cedar hardwood gum Flood levels ( cm ) May 0-2 10 - 40 10 - 20 10 - 40 October 0 0 0 0 Number of Animals Trapped Species Peromyscus leucopus May 0 7 1 3 October 1 1 0 2 Ochro t omys May 0 0 0 3 nuttalli October 0 0 0 1 ANO VA tests revealed an overall significant (p 0.05) difference in total mast production by community and by collection date. The a priori t-tests indicated (1) no significant difference between the spring and autumn collections, and (2) a significant difference between mast production in July and September compared to January and March, and between that of January compared to July. A significant difference was found between mast production at the mixed hardwood stand and the other stands, but no significant difference was found between the cedar stand and the other combined study sites. Apparently flooding may be important in increasing site differences in mast production, but soil type is not. SMALL RODENTS IN THE DISMAL SWAMP 19 Table 3. Mean annual total dry weight of mast collected in 15 collectors, in g/m2, in four Dismal Swamp communities, from September 1979 through July 1980. Values in parentheses are one S.E. Dashes indicate that the species is not found at the community. Species Mixed Hardwood % Maple- gum % Cypress % Cedar % Tupelo 1.60 (1.60) 17.5 28.68 (10.04)- 81.5 7.36 (4.04) 68.3 - Cedar - - - - - - 13.76 (1.17) 73.9 Blackgum 1.56 (0.60) 17.1 2.64 (0.92) 7.4 0.64 (0.20) 5.8 1.40 (0.84) 7.5 Maple 1.16 (0.32) 12.7 1.72 (0.40) 4.8 0.64 (0.32) 5.8 3.48 (0.44) 18.6 Sweetgum 0.92 (0.74) 10.1 2.12 (1.24) 5.8 0.39 (0.39) 3.9 - - Tulip Poplar 2.32 25.4 - - - - -- - Cypress 0.04 (0.04) 0.4 - - 1.75 (0.60) 16.2 - - Oak 1.32 (0.56) 14.5 0.17 (0.17) 0.5 0 0 - - Beech 0.20 (0.19) 2.3 ' ' TOTAL 9.12 (2.44) 100.0 35.16 (9.64) 100.0 10.77 (3.96) 100.0 18.64 (1.56) 100.0 Table 4. Mean dry weight of mast collected in 15 mast collectors from September 1979 through July 1980, in g/m% at four Dismal Swamp communities, by collection date. Values in parentheses are one S.E. Sites are mixed hardwood (MH) , maple-gum (MG), cypress (CY) and cedar (CE) . Dashes indicate that the species is not found at the community. 20 THE VIRGINIA JOURNAL OF SCIENCE continued/ SMALL RODENTS IN THE DISMAL SWAMP 22 THE VIRGINIA JOURNAL OF SCIENCE "Caf eteri an Tests The only fruits consumed during the cafeteria tests were those of tupelo gumi blackgumi oak and maple {Table 5>- Not enough beech nuts were collected for analysis- Negligible amounts of cypressi cedari tulip poplar-i and sweetgum fruits and seeds were consumed! and the mice steadily lost weighti while the control mouse remained close to its original weight- Tupelo and blackgum fruitsi although consumed in quantity^ failed to sustain the normal body weight in the experimental mice- Ir.itial cafeteria testsi using rehydrated tupelo gum fruits-, resulted in large weight losses after two days- There was also a considerable weight loss after two days on a blackgum fruit diet-. Table 5- Cafeteria test data for Peromyscus leucopus- Weight change data represent mouse 1 2 and 3 (control}-, respectively- Weight changes of mice and food are in grams- Control mice were fed sunflower seeds- MAST SPECIES WEIGHT CHANGE (g) PERCENT CHANGE FOOD CONSUMED (g) Tupelo -5.24,-5.76,-2.12 -26. 8%, -29.0%, -9. 6% 12.14, 12.48, 2.57 Cedar -3. 45, -3. 04 ,+0.52 -18.2%, -19.2%, +2.4% 0.00, 0.00, 6.80 Blackgum -3.27,-3.06, -0.20 -17.6%, -16.3%, +1.1% 13.59, 13.83, 4.01 Maple -0.25, -0.09, +0.29 -1.3%, -0.5%, +1.3% 4.80, 3,89, 5.21 Sweetgum -2.72, -2.63, +0.09 -12.3%, -14.4%, +0.5% 0.02, 0.01, 3.07 Tulip poplar -2.26, -2.54, +0.67 -10.2%, -13.6%, +4.1% 0.01, 0.08, 3.27 Cypress -2.05, *, -0.20 -10.8%, *, -0.9% 0.05, * , 2.90 Oak -0.06, -0.50, -0.11 -0.3%, -2.1%, -0.6% 3.02, 3.04, 3.08 * Only enough cypress for two mice. despite the consumption of large portions of these fruits* These results were unexpected because there was evidence that rodents had eaten both fruits in nature- Specifically-, at the maple-gum standi tupelo and blackgum fruits were found which had been gnawed to expose the cotyledon within the hard seed coat- The tupelo and blackgum tests were repeated-, using fresh fruitsi and it was observed that one mouse pierced through the tupelo gum fruit and seed coat to extract the cotyledon-, but the other mouse did not- Both mice ate the fleshy part of the fruit- Each mouse lost over ID percent of body weight after two days- The same pattern occurred with the blackgum fruits when a considerable weight loss was observed after two days- A comparison of the digestible SMALL RODENTS IN THE DISMAL SWAMP 23 portions of the four edible fruits revealed the following: maple samaras are 75 percent digestible! acorns 43*b percent! tupelo gum fruits 13 percenti and blackgum fruits only percent digestible mater i al • Flooding In October ITVT the ground was dry at all four plant communities {Table 2}. In flay! the maple-gum and cedar communities were the most deeply flooded! followed by the cypress stand* In the mixed hardwood stand only an occasional shallow puddle was seen near the periphery of the community* DISCUSSION Information obtained from three separate trapping sessions in the Dismal Swamp trends to support studies which have concluded that species diversity and density of small rodents in woodlands are among the lowest of all North American habitats {Dueser and Shugart 1^75}* Data from Dismal Swamp rodent collections are available from Ifl^S to the present day! and have been tabulated and evaluated by Handley -C 1 ^ 7 T > - One of the more recent collections was his own made in February and June 1^53* During a total of eight nights of trapping! Handley collected 34 Pj_ leucopus! 14 0* nuttal 1 i ! one Rei throdontomys humul is ! one Hicrotus pennsy 1 vamcus-. and three Hus musculus* Comparisons with the present study are difficult! because Handley's methods were not comparable! and he gave no information as to habitat or the size of the area he trapped- However! a similar hardwood swamp was trapped by Getz {l^bll-! who captured six leucopus per 550 fall & winter trap nights* This figure is somewhat higher than my fall and winter findings* Getz also trapped an oak-hickory swamp! and trapped 32 P- leucopus in the same time frame* Peromyscus leucous is the most common and widespread mammal in the Dismal Swamp* It is equally at home in dry or wet situations! in forest and in brush* Populations of this species appear to fluctuate* Handley {1^74} and others have found it in relatively high numbers* However! Dice {1CI40> found compartively few when he snap-trapped in two upland brush areas! five areas in cypress swamp! and two near Lake Drummond in 1^33 and 1^35* The populations of white-footed mice then were evidently as low as in lem-nA0. Populations of white-footed mice also fluctuate by season* In Illinois! population density was highest during the autumn! but alternately declined and increased in the subsequent seasons {Batzli n77}. However! Batzli's floodplain population was more erratic than the upland population- In the Dismal Swamp! more mice were taken in the spring of nTT than in the autumn! especially at the maple-gum stand* In the summer of ITAD! no mice were found! which is in keeping with the tendency of white-footed mouse populations to decline in the summer* Ochrotomys nut t a 1 1 i is similar to the white-footed mouse in many aspects of its ecology and behavior. The golden mouse only occurs in the southern one-third of Virginia! and is much less common in the Dismal Swamp than is the white-footed mouse* Dueser and Shugart {ITTfi} also found it to be c o m p a r a t i v e 1 y rare* In a mesic forest at Oak Ridge! they captured a total of lt>7 P* leucopus in TUTU summer trap nights compared to 12 0^_ nuttalli* Habi tat/flam mal Relationships Vegetation Both mouse species found are nocturnal! territorial! and 24 THE VIRGINIA JOURNAL OF SCIENCE arboreal {Baker One of the major differences between them lies in the importance of microhabitat details- These details appear to be less vital to the white-footed mouse than to the golden mousei Dueser and S h u g a r t have called them microhabitat generalists and specialists! respectively. Both ground cover and shrubs should be dense to support large numbers of these mice- H'Closkey and Lajoie {X^TS}-. who examined the relationship between habitat structure and abundance of P. leucopus i found that mouse populations increased as the vegetation density of the □ — 7 - E. cm layer increased- This was attributed to a combination of factors such as food distribution! nesting sites! and complexity of escape routes. The herbaceous layer at all four Dismal Swamp communities was quite sparse {Dabel and Day 1T77>! its biomass was highest at the mixed hardwood stand {167. T kg/ha>! and lowest at the maple-gum stand {25-7 kg/ha>- Cane {Arundinaria giganteaX is the principal herbaceous component at the mixed hard wood standi but is probably neither tangled nor thick enough to provide an ideal habitat for either species of mouse. Ideal golden mouse sites have a dense ground cover and undergrowth of briers and vines {Goodpaster and Hoffmeister XTSMi Blus ITbbX. The estimated biomass of the shrub layer at the cedar stand was highi IBSl.T kg/hai compared to a low of L>2-7 kg/ha at the cypress stand {Dabel and Day 1^77}. The cedar stand contains a large number of fallen cedar treesi which provide excellent runways for the mice- H'Closkey {nTS} and Getz {XTblX have found a greater number of white-footed mice where there are fallen logs- In addition to runwaysi thick shrubs are essential as protection to small rodents against predatorsi especially raptors {Hiller and Getz 1T77X- The cedar stand contains appropriate shrubs and debrisi which may in part explain why a few mice were found there in Hay and October n?1! {Table 2}. Howeveri because of the larger numbers of white-footed mice found in the maple-gum stand in Hay 1^7^! it is clear that habitat is not the sole factor which influences P. leucopus density. Habitat does appear to exert a greater influence on the 0^ nuttalli populations! which is compatible with Dueser and Shugart {n76}! Goodpaster and Hoffmeister {1HS43-! and Linzey {XHbBX- F looding Although low numbers of mice were found! there is no indication that flooding directly decreases their densities- During Hay! the maple-gum and cedar communities were inhabited by a few mice! but in the dry! mixed hardwood standi none was caught- Batzli {1^77}! who compared populations of the white-footed mouse in floodplain and upland forests! found them to be similar in density at these locations- He observed no detrimental effects of flooding. Swimming tests using white-footed mice {Getz ITb?! Sheppe lUSi Carter 1H7H} indicate that this mouse swims well! especially when it can clearly see its destination- Paschal et al- {ITT^X concluded that inundation was not detrimental to nesting because they found larger numbers of white¬ footed mice in inundated areas of their Dismal Swamp study sites than in dry areas- In addition! they observed mice climbing and nesting in trees- Stah {1H60X studied the vertical nesting distribution of white-footed mice under experimental conditions and found 17 of 2b mice elected to nest at heights of at least 77 cm! compared to 36-5 cm and ground level choices- The nearly prehensile tail of the golden mouse is of great adaptive value to its arboreal existence- In addition! this mouse has a smaller hind foot than the white-footed mouse! which may make it an even better climber {Harper 1H27X- Golden mice are SMALL RODENTS IN THE DISMAL SWAMP 25 known to store seeds in their feeding platforms for future use {Goodpaster and Hof f meister IHSM}. It seems feasible that they could survive flooded swamp conditions i in this study! they swam well when released- Handley -CIHHA} reported that this species is rarely found farther than 3D m from a body of water- Furthermore! the survival rate of this rodent during an east Texas flood! which lasted several weeks! was found to be 64 percent {Packard and Carner 1HL.4}- All other criteria being equal! perhaps the mice choose flooded areas as a defense strategy against predators. Whatever the case! it seems that these rodents are well adapted to flooded conditions! and there is no evidence that flooding reduces their population density! within the Swamp or elsewhere- Food A vai labi 1 i ty The bulk of the food of white-footed mice is the mast of trees and shrubs! but they also consume grains and insects* They need to eat about 30 percent of their body weight each day {Hamilton 1H41>* Food resources are known to influence numbers of white-footed micei and several studies indicate a direct relationship between low mast production and low numbers of mice {e*g*i Jamesoni ITSB}* Similarly! in the Dismal Swamp plant standsi mast production and numbers of white-footed mice both are low* In a study of red oak mast production! Hansen and Batzli {1^7^} considered 173*3g m“5 yr-1 to be a high level! and 14*Dg m” ^ yr”1 a low level of mast production* In this study l*32g m -c! yr of acorns fell! which is only T percent of Hansen and Batzli's low figure* Secher Jensen {1^75} found that winter reproduction in the bank vole {Clethrionomys glareolusl and the yellow-necked mouse {Apodemus flavicollisl failed to occur in Danish forests with low mast production! but did occur where mast was produced* This finding indicates the vital role of mast to forest-dwelling rodents* Getz {IHbll found that white-footed mice were more numerous in dry upland {o ak -h i ck or y> forests than in a low-lying swamp* However! Batzli {1H77> found comparable densities in floodplain and upland forests* Batzli's study area comprised larger numbers of oaks and hickories! in both wet and dry sites! whereas these trees were only found in the drier upland areas of Getz' study sites* Both oak and hickory mast constituted a large proportion of the diet of this species {Getz ntli Batzli 1H7?! fletzgar 1HS5}* Hast stored for future and emergency use is of obvious importance during flooded seasons in a swamp* By contrast! fruits such as those of the red maple {also present in Getz' swamp areal are less likely to be stored! due to their softer tissue andi their potential use is only seasonal {Getz l^bl}* Blus {IHLib} found the three major food items of the golden mouse to be small acorns! poison ivy seeds {Rhus radi cans} and blackberries {Rubus spp*>* Goodpaster and Hoffmeister {l^SM} determined sumac {Rhus spp*>! wild cherry {Prunus spp*>! dogwood {Cornus spp*>! and greenbrier {S m i 1 ax spp*} seeds to be important food items* Poison ivy and greenbrier are found at all four stands in the Dismal Swamp* Blackberries are found along the periphery of the cedar stand* As with the white-footed mouse! the lack of food which can be stored above flood levels for long periods of time appears to be a possible density-limiting factor for golden mice* The literature on golden mice implies that this rodent is not such a generalist in its feeding habits as is the white-footed mouse* There isi for example! no indication that the golden mouse eats insects! as does the white-footed mouse* In this study! maple fruits fell at all four standsi but only 26 THE VIRGINIA JOURNAL OF SCIENCE fell at the cedar and maple-gum stands! which may have contributed to the higher number of white-footed mice found at these stands in May! and to the presence of golden mice at the cedar stand in flay. Acorns were only found at the mixed hardwood standi and in small quantities. The size of an acorn crop varies considerably every yeari perhaps our collections were made in a year of low acorn production. Beech nuts are also ediblei but too few were collected for the cafeteria tests. Like acorn production! the amount of beech mast varies annually. Secher Jensen {ITTSli who measured beech nut production in a Danish forest! characterized low mast production as less than S g m yr” • With regard to mast production quantified in our study! £ pr i ori contrasting revealed significantly less mast at the mesic mixed hardwood stand than at the other stands. Howeveri due to the enormous fluctuations in acorn and beech nut production! a ten year studyi for example! might well show a reversal of the situation! with mast production of the mixed hardwood stand becoming significantly greater than that of the other stands. A comparison of Tables 5 and 3 shows a potential correlation between the slightly larger numbers of white-footed mice at the maple-gum and cedar sites and the large total amounts of mast produced there. However! closer examination reveals little cause- and-effect relationship between mast production and mouse numbers. Maple fruits have already been mentioned as a possible! but temporary! cause for the higher numbers of mice at the cedar and maple-gum stands. The large amount of tupelo gum mast at the maple-gum stand is only a potential food source. A slight weight loss on an exclusive diet of a new food is not totally unexpected! but the weight loss of the mice fed gum fruits was cons i der ab 1 e- Because of these weight losses! it seems that the energy expended by gnawing the tough seed coat to obtain the cotyledon outweighs the energy derived from that cotyledon- The mouse that did not attempt to gnaw the seed lost no more weight than those that did- The fleshy portions of the fruits are evidently indigestible! possibly because of the waxy component to the covering. In addition! if the gum fruits were used as food! one might have expected to find a greater density of mice in October than in May! especially at the maple-gum stand! which was not the case- It is worth noting the comparative amounts of digestible material in the four edible mast species- The amount in the gum species is negligible when compared to oak and maple! given the lack of digestible value of fleshy parts of the gum fruits- In summary! there does not appear to be one single factor which limits the diversity and density of small rodents in the four Dismal Swamp plant communities- Diversity is predom inant ly affected by the flooded conditions which prevail throughout most of the Swamp. However! if flooding were the only factor limiting diversity! a greater variety of species would be expected at the mixed, hardwood stand. This was not the case! so other factors appear to be involved. ACKNOWLEDGEMENTS This paper is based on a Master's thesis submitted to the graduate faculty of Old Dominion University by the first author- The research was supported in part by a grant from the Virginia Academy of Science- The assistance of the following individuals is gratefully acknowledged: Marta Gomezi Allen Plocheri George Farrowi Joe Atchue! Becky Cintron! Jerry Sourbeer! John Breidlingi Paul Homsher! Rachel Breidling! James Mattai Lytton Musselmani and Gerald Levy! and Susan Bonner. Appreciation is also given to the staff of the Great Dismal Swamp National Wildlife Refuge- SMALL RODENTS IN THE DISMAL SWAMP 27 LITERATURE CITED Bakeri R- H- lUfl. Habitats and distribution- pp- T8 - 15b- In J- A- King led-} Biology of Peromyscus {rodentia}- Spec- Publ- Amer- Soc- flamm- 2: 1 - ST3- Batzlin G- 0- 1177- Population dynamics of the white-footed mouse in floodplain and upland forests- Am- Midi- Nat- T7: 18-32- Berkelyn E-! and D- Berkely- l^b- flan and the Great Dismal- Va- J- Sci- 27: 141 - 171- Blusi L- J- ITbb- Some aspects of golden mouse ecology in Southern Illinois. Trans. Illinois St- Acad- Sci- ST: 334 - 341- Cartern J- L- n7T Swimming as a determinate to immigration for small mammals in Coastal Virginia- -CUnpubl- Plaster's thesis! Old Dominion University}- S8 p- Dabeli C- V-! and F - P- Day! 1T77- Structural comparisons of four plant communities in the Great Dismal Swamp! Virginia- Bull- Torrey Bot- Club 104: 352 - 3b0- Dice! L- R- 1^40- Relationships between the wood-mouse and the cotton mouse in eastern Virginia- J- flamm- 21: 14 -23- Dueseri R- D-! and H- H- Shugarti Jr- 1^78- fl i cr oh a b i t a t s in a forest floor small mammal fauna- Ecology ST: 8 ^ - 1&- Getz i L- L- 1 1 b 1 - Notes on the local distribution of Peromyscus leucopus and Zapus hudsoni us- Am- Midi- Nat- b S : 48b - 4eH- _ _ 11b7- Responses of selected small mammals to water. Conn- Univ- Occas- Papers Biol- Ser- 1: 71 - 81- Goodpaster! Id- bJ-! and D- F. Hoffmeisteri 1T54- Life history of the golden mouse! Peromyscus nuttal 1 i ! in Kentucky. J- flamm- 35: lb- 27- Hamilton! Id- J- Jr- 1T41- The food of small forest mammals in the eastern United States- J- Hamm- 22: 250 - 2b3- Handley! C- 0- Jr- 1^48- Habitat of the golden mouse in Virginia- J- flamm- 2T: 2^8 - 2^- _ _ _ l^T flammals of the Dismal Swamp - a historical account- pp- 2^7 - 357- I_n P- Id- Kirk! Jr- ■Ced} The Great Dismal Swamp. Univ- Press of Virginia! Charlottesvi 1 le- 427 p- Hanseni L- P-! and G- 0- Batzli- 1T7T Influence of supplemental food on local populations of peromyscus leucopus- J- flamm- b0: 335 -342- Harper! F- 1T27- The mammals of the Okefenokee Swamp region of Georgia. Proc- Boston Soc- Nat- Hist- 38: m - 3Tb- Jameson! E- Id- Jr- 1 ^ 5 3 - Reproduction of deer-mice -CP eromyscus maniculatus and P- boyleil in the Sierra Nevada! C a 1 i f o r n 1 a • J- flamm- 34: 44 - 58- Lack! D- lc154. The natural Regulation of Animal Numbers- Oxford Univ- Press London- pp- 343- Levy! G- F-! and S- Id- Idalker- IT?1!. Forest dynamics in the Dismal Swamp of Virginia- pp- 101 - 12b- In P- Id- Kirkr Jr- led-} The Great Dismal Swamp- U nTv - Press of Virginia! Charlottesville- 427 p- 28 THE VIRGINIA JOURNAL OF SCIENCE Linzeyn D. lil - nbfl- An ecological study of the golden mouse! Ochrotomys nuttallin in the Great Smokey Mountains National Park. Am. Midi- Nat- 71: 32 D - MS. M'Closkeyn R. T. 1T75. Habitat dimensions of white-footed micei Peromyscus leucopus. Am. Midi. Nat- 13: 15fl - lb7- M'Closkeyn R- T- and D- T. Lajoie- 1^75. Determinants of local distribution and abundance in white-footed mice. Ecology 5b: 4b7 - 472. Metzgari B. ITS 5. Notes on mammals of Perry Co.'i Ohio- J- Mamm- 3b: 101 -IDS. Millern D. H.n and L- L- Getz- 1177. Comparison of population dynamics of Peromyscus and Clethrionomys in New England. J. Mamm. 5fl: 1 - lb- Oaksi R. <3. Jr.i and D- R. Uhitehead- 1T7T. Geologic setting and origin of the Dismal Swampi southeastern Virginia and northeastern North Carolina. pp. 1 - 24. I_n P - UJ - Kirkn Jr. -Ced-J The Great Dismal Swamp. Univ. Press of Virginian Charlottesville. 427 p. Packard! R- L-n and H- Garner. Iclb4. Arboreal nests of the golden mouse in East Texas- J. Mamm- 45: 3\?1 - 374. PaschaL J. E. Jr.i D- E. Sonenshinen and J- H. Richardson. n7T A simulation model of a Peromyscus leucopus population in an area of the Dismal Swamp. pp. 277 - 2cib. I_n P- U- Kirkn Jr. -Ced-J The Great Dismal Swamp. Univ- Press of Virginian Char lottesvi 1 le- 427 p. Secher Jensenn T. 1^75- Population estimations and population dynamics of two Danish forest rodent species- Vidensk. Meddr- dansk naturh- Foren- 13fl: b5 - fib- Sheppen Id. nbS. Dispersal by swimming in Peromyscus leucopus- J- Mamm- 4b: 33b - 337- Stahn C-D- nflU- Vertical nesting distribution of two species of Peromyscus under experimental conditions- J- Mamm- bl: 141 - 143- Stewartn P-C- l^T Man and the Swamp: the Historical Dimensions- pp. 57 - 73- I_n P. Id- Kirkn Jr- -Ced-J The Great Dismal Swamp- Univ- Press of Virginian Char lottesvi 1 le- 427 p- Idhiteheadn D- R-n and R. (3- Oaksi Jr- lci7c|. Developmental History of the Dismal Swamp- pp- 25 - 43- I_n P- Id- Kirkn Jr- -Ced-J The Great Dismal Swamp- Univ- Press of Virginian Charlottesvi 1 le- 427 p- Virginia Journal of Science Volume 34, Number 1 Spring 1983 Determination of the Smallest Size of a Sample Selected From A Finite Population for an Attribute with a Given Population Proportion N. N. Mikhail, T. L. Weaver and L. P. Lester Liberty Baptist College Lynchburg, Virginia 24506 ABSTRACT Mikhail, Lester and Weaver (1982ajhave shewn that the distribution of the sample percentage is very accurately approximated by a beta distribution, B (a,8)> using a two-moment graduation. We have found that the parameters of the beta distribution, a and 8, depend only on the finite population size N, on the sample size n, and on a fixed value of population proportion. In this paper, we have used the results illustrated in Tables 1.1,..., 1.7 to determine the smallest size of a sample drawn without replacement from a finite population for an attribute with a given population proportion. A comparison of Wolde-Tsadik* s (1980, The American Statistician) results with our results is included. 1 . INTRODUCTION Mikhail, Lester and Weaver (1982a) have shown that in the case of a given finite population, the sampling distribution of a sample proportion is very accurately approximated by a beta distribution using a two-moment graduation. Furthermore, we have found that the beta model depends only on the finite popu¬ lation size N, and the sample size n, but not on the value of the population proportion P. It is noticed that the value of P is always constant and equal to 3/10, and the computation for 8^ (skewness) and 82 (kurtosis) for the actual distribution of sample proportion and for the beta distribution are depicted in Table 1, for different values of N and n, and P = 0.3. The chi-square model is found to fit very poorly. 2 . THEORY Given a sample of size n drawn without replacement from a finite population of size N, the first two moments (Mikhail, Lester and Weaver, 1982a) of the sample proportion are given by: E(p) ■= P Vac(b) P(l-P) /N-n, n Vr 2.1 The Beta Model For a sample proportion of p of a sample size n drawn without replacement 30 THE VIRGINIA JOURNAL OF SCIENCE from a finite population of size N, the beta distribution with parameters a, and 6 is given by: P~ B(a, 0) (2.1.1) with the probability density function (p.d.f.) , r(a+0) o-l „ .,0-1 f(p) ‘ r (a)7(i3) p <1_p) Where T(a) s° a-1 -x , / x e dx a > 0. The first two moments of p are then (2.1.2) E(p) ■ jj ■ o/(a+0) Var(p> ■ wfapir ' >‘(l-W)/(a+e+l) . (2.1.3) Using the first two moments of section 2, we have a H(n-D P (N-n) . W(n-l) (N- (1-P). (2.1.4) For most of the values of N, n, and p, a and 0 are integers, a fact which makes the usage of the beta model very practical for sample size n. The values of 01 and 02 for the beta model are given in column II of Tables 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, and 1.7. 2.2 Chi-Square Model Given the sample percentage p for a sample size n drawn without replacement from a finite population of size N, the scalar multiple of chi-square distribu¬ tion with parameters X and C, is given by, P ~ C)U or p/C - xjj where X stands for the degrees of freedom (d.f.) The p.d.f. for this model is , 4 - 1 .-xl/2 f(x2) * -X7T7M fe'J* i Oix i r'1 r(|) with p/C as a chi-square variable. (2.2.1) (2.2.2) The first two moments of p are E(p) ■ CX Var(p) - 2C^X, (2.2.3) Using the first two moments of p in Section 2, we have C X (2.2.4) The values of of Tables 1.1, 1.2, (skewness) , and 0_ ; 1.7, it is clear th Si and 02 for the chi-square model are given in Column III 1.3, 1.4, 1.5, 1.6 and 1.7. From the values of 0. = h^/y« = y^/l^ (kurtosis) in Tables 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, at the population proportion P = CK3, gives a very good fit 3/2 DETERMINATION OF SMALLEST SAMPLE SIZE 31 of beta distribution to the actual distribution of sample percentage. The values of (3. and in Column III for chi-square model is obviously showing a poor graduation to the actual distribution of sample proportion. Accordingly, the estimated values of a and $ are reduced to the form of a - 0.3 N(n-l) / (N-n) g - 0.7 N(n-l) / (N-n) . 3. DETERMINATION OF THE SAMPLE SIZE Consider the situation in which sampling units in the finite popula¬ tion of size N are divided into two mutually exclusive classes: class 1 con¬ sisting of units possessing the attribute under consideration with a given pop¬ ulation proportion P, and class 2, consisting of those not possessing it. Here we select a sample of size n from the finite population without re¬ placement. From the beta model in section 2, we have: a - 0 . 3N (n-1) / (N-n) 0 - 0. 7N(n-l)/ (N-n) , for which p.d.f. of beta distribution is determined. From this beta distribution, we select the value of the sample proportion p^ ^ corresponding to the upper 5 percent level as an estimator of the populationproportion P of the units possess¬ ing the attribute of class 1. This estimated value P = pQ , will be regarded FOR ACTUAL TABLE 1.1 DISTRIBUTION AND DIFFERENT MODELS, N = 50 VALUES OF 0 AND 0 1 2 1 II III s ■ s EH SS 1 . 15E-01 4 . U0E--01 1 . 7 1 E + 0 0 p = .3 B2 S3 2.75E+00 2.71E+00 5.57E+00 s * 10 B1 ss 3 . 65E-02 2. 13E--01 7. A2E-01 p * . 3 B2 ss 2 . 87E+00 2 , 88E + 00 4 . 14E+00 s = 15 B1 ss 1 .23E-02 1 . 32E-0 1 4.44E-01 p * .3 B2 ss 2.91E+Q0 2 . 93E + 0 0 3.67E+00 s - 20 B 1 ss 2.70E-03 8 . 78E-02 2.86E-01 p * . 3 B2 ss 2.92E+00 2 . 95E + 0 0 3 . 43E-*-0 0 s = 25 B1 ss 0 . 00E+00 5.97E-02 1 .90E-01 p * .3 B2 ss 2 . 93E + 0 0 2.97E+00 3 . 29E + 0 0 s = 30 B1 ss 2 . 70E-03 4 . 04E-02 1 . 27E-0 1 p * .3 B2 ss 2.92E+00 2.98E+Q0 3. 19E+00 TABLE 1.2: VALUES OF 0 AND 02 FOR ACTUAL DISTRIBUTION AND DIFFERENT MODELS, N = 75 I 11 III S - 5 B 1 SS 1 .28E-01 4 . 08E-01 1 . 7 7E + 0 0 P * .3 B2 ss 2 . 75E+00 2 . 70E+0 0 5.65E+00 S = 10 EH ss 4 , 92E-02 2 . 26E-0 1 8.20E-01 P = . 3 B2 = 2.88E+00 2.87E+00 4.23E+00 s * 15 B1 ss 2.38E-02 1 . 48E~ 0 1 5. 05E-01 p ■ . 3 B2 ss 2 . 92E + 0 0 2.92E+00 3.76E+00 s » 20 B 1 ss 1 . 18E-02 1 . 05E-01 3 . 47E-01 p ■ . 3 B2 ss 2.93E+00 2.94E+00 3.52E+00 s = 25 B1 ss 5.29E-03 7.81E-02 2.52E-01 p * .3 B2 ss 2. 94E+00 2 . 9AE+00 3 . 38E + 0 0 s * 30 B1 ss 1 . 76E-03 5.93E-02 1 . 89E-0 1 p * .3 B2 ss 2.95E+00 2 , 97E+0 0 3.28E+00 s = 35 B 1 SS 1 .89E-04 4 . 5AE-02 1 . 44E-01 p « . 3 B2 SS 2 . 95E+00 2.98E+00 3.22E+00 Cj » 40 B 1 ss 1 . 89E--04 3 . 52E-02 1 . 10E-01 p a .3 B2 ss 2 . 95E + 0 0 2.98E+00 3. 17E+00 s a 1+5 B 1 ss 1 . 7AE-03 2 . 70E-02 8.41E-02 p a . 3 B2 ss 2 . 95E+00 2 . 99E+00 3 . 13E+-0 0 s a 50 B1 ss 5 . 29E-03 2 . 03E-02 6 . 3 1 E - 0 2 p a . 3 B2 S3 2.94E+00 2.99E+00 3 . 09E + 0 0 * s = sample size (n) , and 1.29E+01 = 1.29 x 101. 32 THE VIRGINIA JOURNAL OF SCIENCE "C cn 93 cd T! w 93 a o ■c co -c CO -= CO — CO ■o CO ~ CO *D CO TJ CO •c CO -0 CO ~c co n Si y I! H ii t: H ): ii ii ii ii H H !? ii Ii ii jj H « ii H !! 1! B ii o — 1 > - -si -si CS O' - cn - CJ\ - -P - P • CO CO SO ro r-4 CO —1 03 co C* = co cn CO ° co (Jz C*i G CO CB CO o CO cn CO o CO cn CO G CO f n CO O co cn DO m a? 23 23 23 23 rr s w 5? * 5? 2? 2? 2? rr- 2? 2? 2? 2? OC 2? 2? 23 23 2? a? 5C 23 2? 23 oa N5 *-* ro ’-*■ ro ro ro i— ro '■-1 ro ro *— 1 ro ro H1 ro !“*■ ro ro i-+ ro i-1 ro 5 CO ii IS ii li ii ii ii fi ii H Ii ii u ii ii ii B ii ii ii ii ii n B ii ii u ii Si Ii o ro ro Cm ro w ro J-4 ro to ro -P ro CO ro ►* ro CO ro cn ro ro ro W ro cn ro ^ 1» < > 'Co>C G >0 _l >0 o* g H* >0 1— ' 'G i _* >C CO >0 F-4 G G G o G -si G o oo cn -si CO z o (— O' !-*■ O' -si O' >0 O' -0 O' *-• O' F-4 O' CD cn cn f -si ro ro oo 00 cn p »— ! m p p p p p p p m rn p m fT! rr PS Pi PI rn m p p p pi p p p p p p p p o co + | + i + i + J + i + i + I + ! + i -f ! + i + I + i + i + i o G o o © o o o G O G ro G G G O G G © O o o o G o o o o G G ~n o G ro G CO G W G U" G -P G G G p G CO G « G U: G ro o ro g ro G wen i j m 30 03 m CO *-*• ro »-*• N3 H4 ro ro 2, N3 ro CO ro CO ro -p ro cn ro CK ro oo ro 9 1 , 2 , ro ro p z —l > o o g co g o >c o >c P >C G sQ O' G P G cn G GC G -si G i— *■ g cn oo l pi -j z (31 g fO >0 M- G p >0 ro >0 oc >c ro CD “sj CD CD -si -si O' O' o P p H O' O' CO c= ro o p !Tj P m m p rn p PI m rr in PI P! p p p P p p P PI P p p p p p p p HH o 03 + j 4- Y V i i + i + i + 1 + i + I 4- i + i + i + i + i + i m ro G G G c o G © G G O G G G G G O G G G G G G o G o o G o G O co ° G ro G ro ° ro ° ro o ro ro g ro G ro G ro G ro c 1-1 G !-*■ G *■* G 1-^ O 30 co co co p CO c* c* O' CO -si CO 'C u - CO H» CO i— » w ro CO ro CO CO co cn P CD cn t— *■ z ii o f— *■ © © © G © ro H* “Si *■* -p H4 r o -p ro -sj ro -p CD cn ■vj CD CO ro P O' -si —i cz cn JT ■F oc cc NO >6 ro H* -F to i-* t-1 cn Csj G ro CO -si © p -si 0D G G F— f o J3 rn rn m JT! rr JT: m rr m JT: JT: p m ?rj p p JT rr rn m p m m m rr p P P P P H o 1— + i -f i V ! + i V i -f i V i + i V V i + Y + i V 1 + 1 + + G o g o © G © o G G © o G G G G © G G G G G G o G o G G G G G ro g ?0 g ro G ro G fO G ro G F* O 1— *■ © J-4 G r-4 G G G )-*• G I-4 G G — - CO ~C CO "O CD 93 CD 93 CD “C CD 9C CD 93 CD 93 CD 93 CD ~ CD 93 CD¬ 93 CD 93 CD 93 CD “C CD 93 CD 93 CD 93 CD ii li B ii fi u ii n b B li H ii ii ii ii ii B ii S'. S i! H ii R i! ii ii B B n ii H ii G - G - CD OD - -sj - -si O' O' cn cn - P . P . CO . co - ro - ro - . i-i . CO cn co g co cn CO ® CO cn co g CO cn CO G CO cn CO G co cn CO o CO cn CO o co cn co o co cn CO G CO -n- 23 rr 2? 2? 2? a? 2? 23 23 2? 23 2? 23 23 23 tr 23 23 23 23 23 23 23 23 23 23 23 23 23 23 23 23 2D w 23 rr 23 23 ro h* ro m- ro •“* ro h* ro h ro h ro !-» ro !-*■ ro i-*- ro ►-* ro )-* ro ro ro HMH ro »-* ro H4 ro ro »-* n 1! ii Ii ii B ii R H 0 R U ii ii ii li ii B R ii S R H B B li 11 H R B ii H B H R ii li ii ro 2 8, 9 1 , CO ro g ro nroo ro CO ro CD ro h 1 ro ro ro CO ro cn ro CO ro »— 1 ro ‘ ro r o ro C*i ro O' ro * G cn G O' G -si G ro g p g ro G -J G O' G c.n G cn g G G -si G ro G G O' G P G O' CD P.J -si P -J GOOPCDcncDcncDcocDcn 00 cn od P -si > c G -si cn -j -si O' P O' -j cn O' p ^ ro G 03 O' cn o H P P P P P P p p m p nn p p p p P P P P p p p p p P p p p p p p p p p p p p P + f + i + 1 •f i + I + 1 + i + 1 + i + i + 1 + 1 + i + ! + i + i + 1 + i + 1 o o o o o o G G G t— * o o G G o o G o o o o o o o G o o GOO GOG o G o G o o CO o p G P G cn g g o cr. G p G P O G CO G CO o CO G CO G ro o ro g ro g ro G ro G '■* ro h rj h ro h ro m- ro ro ro ro ro ro ro CO ro CO ro co ro p ro cn ro O' ro -si ro g 1 , p 1 , 2 ro ro ro p G Hr G CO G cn G -si G g G CO G O' G o G P G G G O' G p G PG-sIGCnGfOGO' CD CO -si G cd g cn G cn G -si G ro G G G G t— * CD C" CD CD CD CO -si ro -j CO O' O' cn -sj p ro h P O' G c; CS H p p p p p p P p p p p p p m P P P ni P p p p p p p pppppppp p p rn P m J— 1 + i + i + i + i + i + i + i + i + t + i + i + i + 1 + i + i + i + i + i ■f i G G G O O G G o o GOG o © G G G G G G o G o G o o o o o c G G G G G © G c G ro ® ro g ro O ro g ro g ro G fO G ro G ro G ro g ro G ro o ro o ro g ro G 1— G F- 4 G j-4 G CO CO CO P co p CO cn co O' CO -si G CO >-* CO H* CO CO CO ro CO ro co co CO P CO U: P oc cn c; O' G I-*- G -si C; P G ro h h i_s -F H4 G !_* ro ro P ro CO G CO cn p h* O' o 00 O' CO -sj -s! •X cn CO O' OD —J G SO 00 G O' h1 0- ro G P G OD G cn ro O' O' ro O' 00 H*- -si CO !-*• -si cn -c- ro -si ro ro p P P P P P p P P p p p p P P m m m P rr m p nr, p m p p m rn m m rr rr m p fT: rr m V- ! + i + i + 1 + i + i + i + i + i ’+ i + Y V 1 ’+ Y + I + Y + i + 1 + 1 + 1 + + o O G O G G c O G O G G G G G G o G G O G G o o G G o O G O G G G G o o G © G ro g ro o ro G ro o ro g ro G ro G rc G f"* G (-* o H* G *-» G G 1-*- G I-* G 1— * G G H4 ® G TABLE 1.4: VALUES OF 6 AND 8 FOR ACTUAL DISTRIBUTION AND DIFFERENT MODELS, N * 150 TABLE 1.5: VALUES OF 3 AND 3 FOR ACTUAL TABLE 1.6: VALUES OF 3x AND 3z FOR ACTUAL DISTRIBUTION AND DIFFERENT MODELS, N *= 250 DISTRIBUTION AND DIFFERENT MODELS, N * 500 DETERMINATION OF SMALLEST SAMPLE SIZE 33 WOrtOHOHOnOHOHOJOJ^ OQ030 oooooooooooooooooo=> = ^--' + + !2R555S2SSSSS2tS2a2 5 = 3 = S£H5«»5.»» > d- d- M CJ W r-J w M 1-t co coo^roooro^-roMJCODCODco^ro^co ^OT- r-t O' CM CM i H t-i CM A A o CO CO - a a to a 1 + 1 + UI UJ Ui UJ r- o ® ± O' o O' t-i CM »-• CM it a « h HCMHCM A A A A o CO o CO * -ID - HUBS CO CL CO A I + Ui UJ O- CD CO o © CM a a t-i CM A A UJ UJ o GO -© O' >0 CM 8 B t-i CM A A O CO -o - a h co a O CO c- - a a cn a UJ Ui Ui UJ c- 00 in ^ CM O' CM O' HO CM d* CM H « H H tH CM t-i CM A A A A o CO o CO © - O' - B n H H co a, CO a. Ui UJ Ui ID O' o ± O' CO CO CM CM B H H cm t-i A A A * CO * * - St 11 a h CO a CO o CO o CO 0000 + 1 + 1 Ui UJ UJ UJ O' C- O' .+ O' CM O' ® CM CM CM t-i ii a a a CM wCMrl A A A A co st co * - * - st H B ii Ii o_ co a cn o co o o + i UJ UJ O' Is- 0' * CM *•« 11 a CM ’-1 A A CO * - * ii B A CO UJ UJ O' Is* O' H CM t-i CM a b a CM t-i CM AAA CO Sc CO - * - HUB A CO A 0 T-I O *-* O t-4 O T-* O T-I 0 CM 0 CM 0 CM 0 CM 0 CM 0 CM 0 CM 0 CM 0 CM C3 o 0 O O 000 0 0 0 9 0 0 0 0 O 0 0 000 0 0 0 O 0 O 0 0 1 + 1 + I + I + 1 + 1 + 1 + 1 + 1 + i + i + 1 + 1 + 1 + 1 + j— i Ui Ui Ui UJ Ui UI UJ Ui Ui UJ Ui i-Ls Ui Ui UJ Ui Ui Ui Ui Ui Ui UI UJ Ui UI Ui Ui Ui Ui Ui r-H 0 ID H ill iiJ rt C" 0 0 CM O' CD d- d- r- CM M3 0 CM A ^ r^. M3 M3 >c D D dr 0 ± r—i 0 CO CO >0 -> d O' CO D CM t-i T-i M3 1—1 0' H M3 r-i >0 0 c- 0 0 0 d- O' 0 D O' d d CO 2 . 3. 1 . 3 . 1 . 3 . 1 . CO O' CO c- CO 6. 3. D CO * CO Jr co *r\ «« CM CO CM CO i-C o T~* O CM 0 CM 0 CM 0 CM 0 CM 0 CM 0 CM O CM 0 CM 0 CM O 0*4 0 CO O CO 0 0 ■ — ■ 0 0 O ® o OOOO 0 0 0 0 O O 0 000 0 0 0 O 0 0 0 0 0 1 + 1 + 1 + 1 + 1 + 1 + 1 + 1 + 1 + 1 + 1 + 1 + I + 1 + I + j— i Ui Ui UI UJ Ui Ui Ui Ui Ui Ui Ui Ui Ui Ui UJ Ui Ui Ui Ui Ui Ui Ui Ui Ui ui Ui Ui Ui Ui Ui h-4 ± iiOO'WN'OM)Nlffl®CDa) O' MJ O' D O' r-i O' d- O' tM O' CM O' 'OSI'J 0 ± ,8 2 9 ± O' t-i O' r- O' r» O' 0 O' ID O' A O' D O' CO O' O' D O T-I 0 2 , 2 , tH CM CD CM 'O CM Jt CM CO CM CO CM CM CM 2 2 , 1 , '3 1 , CM T-I CM CM O' CO A CO CM 0 CM 0 2 0 2 0 CO 0 CO 0 CO 0 CO 0 CO 0 ± 0 d- 0 D 0 D 0 di¬ 0 ± 0 O 0 0 O 000 0000 0 0 0 0 0 0 0 000 0 0 0 0 0 es 0 0 0 i + I + 1 + i + 1 + 1 + 1 + 1 + 1 + 1 + 1 + I + ; + 1 + 1 + UJ Ui UJ UJ UJ Ui Ui UiUiUiUlUIUiUiUJ Ui Ui Ui Ui UJ Ui Ui U Ui Ui Ui Ui UI Ui Ui 0 CD >0 ± O' M3 M3 C- ;+ C- Is- © M3 A :+ A ID CD d- O' 0 O' A O' A O' 0 O' Jt O' 00 CD O' O' M3 O' 0 O'O'O'DO'O'O'QO O' 0 O' t-i O' A O' O' O' O' O' A O' t-i O' M3 CM CM CM t-i CM T-i CM M3 CM J- CM CM CM H CM H CM ID CM T-» CM H CM t* CM H CM ID CM If B s B B B H y B B H U II U B II B fl I! ii n R a fl H a a fl II a CM T-I CM T~< CM T-I CM tH CM t-I CM ■H CM vH CM H CM ■H CM T-I CM iH CM T-I CM 'T—t CM _i CM A A A A A A A A AAA A A A A A A A A A A A A A A A A A A A 0 ro 0 CO 0 CO 0 CO 0 ro 0 CO 0 CO 0 CO 0 ro * CO * ro St ro St CO St CO St CO H CM CO - d- ID - M3 - I'¬ - A * O' * * * - SC St ■ St St ii H II il B (1 R H U fl B ii ll B B fl fl 1! Ii II H H fl fl fl ii il a II a A A A A S P A A AAA A A A A A A A A A A A A A A A A A A A . 99E + 0 0 2.99E+0Q 300/89 700/89 0.533 34 THE VIRGINIA JOURNAL OF SCIENCE X cn x ■Si -c cn X cn X cn X cn X cn X cn X cn X cn X cn X cn X cn X cn X cn o 6 ii li li 1! li li ii ii H 1! R li H If H H B i B i B B B B B H B R 8 CO * * * . * * . * >0 . CD . -si « o . cn . F . w w , h4 3D W * W * Cm * Cm * Cm * Cm * CO © w © w © w © Cm © W o W © w © Cm © i — i CD 3? w 0? w oc a? c 'C >0 w >0 -O >0 h* 'C -si >0 F >c w >0 Cm 'C O' >0 o 'O W >c O' o Cm >0 cn 00 F ° >0 F c >c >0 o >c CD >c tfl >0 U 'C -si CD U CD CD CD cn CD © -sj CD O' W F >0 CD t-4 hi aj m rn m m m m m rn m m m m m m m m m m m m m m m m m rn m m m X + 1 + 1 + 1 oooooooooooo® WcUicUioUoUOUoU I + I o W © © © W 1 + © ro i + i o o © WON) W H* W »-» W W W W W W W W W U W Cm W 04 W F W U5 W O' W >0 W ►* W W >c “Si >c CD >0 o >c .4 .9 .2 >0 -si >0 © >0 F sQ sQ 'O O' >o cn >o >o >o w >0 W GD cn >c >c cn vC t-4 >0 © >0 Cm c © >0 3 8 2 ODCnCDW-sICDO'CDCn O' Cm O' cn © ►H m rn m m m m m m m m m m m m m rnrnrnmmmmmmm m m m m m + i + i + i + i + i + i ■T i + i + I + i + i + i + i + i + i © © © © © © © © G © © © © o © © o © © © © © © © © © © o © © ~ w © w © ro © ro O w © ro © 0 2 w © w © w © W © W © ro © H4 © H4 w cn Cm cn w O' co O' C*i -si w CD CO >c w H4 3 1 3 H w H w W W Cm Cm F F nC © w o -si o ro OD cn H4 F H4 F H4 © «-*• w ro F ro -si Cm ro F G CK cn Cm W CD >c vC F C" © IS\ M4 O' Cm h4 F cn cn -s! >0 F W O' -sj GD F F cn f*0 'G CD -c cn m m m m m m m |T. m m m m m m m m m rn m n rn m m m m m rr m rr — r— + i + i + i + \ + i + i + i + 1 + i + 1 + 1 + i + i + i + 1 © © © © © © © O © © ©■ © © © © © © © © © © © © © © o © © © © © w © w o ro © ro © w © ro © w © H4 OH4© I-4 © H4 O H4 © 1 © H4 © H* ui ro vo 00 K> O' "J VO © VO Ui O' © O o t— * o © 3 © --4 O' Ln l>> 3 DETERMINATION OF SMALLEST SAMPLE SIZE 35 as the given population proportion for the selected sample units from the given finite population. These sample units will constitute the smallest size n of^ the sample drawn without replacement for a given attribute with a proportion P. In Tables 2.1 and 2.2, we give the values of sample size n selected from a finite populations of sizes N * 20 and N = 100, for P > 0.5, for units possess¬ ing the attribute of class 1. For any values P < 0.5, we will consider the units not possessing the attribute of class 2 with proportion Q = 1 - P. 4. COMPARISON OF RESULTS Wolde-Tsadik (1980) has determined the sample size based on the maximum length of the interval estimator of the correlation coefficient p when the parent population is normal. He used David’s Tables (1938) for this purpose. He also compared this method with two approximations. The first is that of Abramowitz and Stegun (1965), which is an approximation for the universe F function; the second is the Z-transf ormation of Fisher (1921). The results of Wolde-Tsadik indicate that the first approximation based on the maximum length of the interval estimator of the correlation p is as good as the second approximation and both of them are superior to Fisher's Z-transf ormation. Mikhail, Prescott and Lester (1981), have shown that under normal assumption, Fisher’s Z-transf ormation departs from normality very significantly, even for | p | ^ 0.3. This will explain why Fisher's Z-approximation in Wolde-Tsadik' s (1980) results is inferior to the other methods. Our method for the determination of the smallest size of a sample selected from a finite population of size N for an attribute with a given population pro¬ portion is based on the fact that distribution of the sample percentage is very accurately approximated by a beta model with two parameters, a and 6, which de¬ pend only on the population size (N) and the sample size (n) . Tables 2.1 and 2.2 illustrate the usefulness of this by listing a, 8, P and n for two population sizes, showing the interdependence of n and P. Our method is clearly superior for the following reasons: first, Wolde- Tsadik used an infinite population while we are using finite populations which is more practical especially in biomedical experiments. Secondly, tables for beta distribution can be calculated easily for any a and 8 to provide tables similar to 2.1 and 2.2 with adequate information about the sample sizes selected from a given finite population. In the case of Wolde-Tsadik, more extensive calculation is involved to reach the same results. 5. CONCLUSION The values of 8^ and Tables 1.1 - 1.7 show that the distribution of the sample percentage is very accurately graduated by a beta model. The fact is used to determine the smallest size of a sample drawn without replacement from a finite population. The reliability of the sample sizes given in Tables 2.1 and 2.2 depend on the prior knowledge of the population proportion of the attribute under consideration. Tables similar to 2.1 and 2.2 can be easily calculated for different values of N and n using a beta distribution. The more knowledge one has of the population proportion of that attri¬ bute, the more dependable the values of n given in Tables 2.1 and 2.2 or other calculated tables will be. This makes our method relatively well suited to quality control analyses where the theoretical population proportion is arbitrarily fixed. References Abramowitz, M. , and Stegun, I. A., editors, (1965), Handbook of Mathematical Functions, New York: Dover Publications. David, F. N. , (1938), Tables for the Correlation Coefficient, London: Cambridge University Press. Fisher, R. A. (1921), On the Probable Error of a Coefficient of Correlation Deduced from A Small Sample. Metron 1, 1-32. 36 THE VIRGINIA JOURNAL OF SCIENCE Mikhail, N. N. , Lester, L. P., and Weaver, T. L. , (1982a), Sampling Distribution of Proportions and Percentages with Applications for Finite and Infinite Populations. Will appear in Virginia J. Sc., Vol. 33, No. 4. Mikhail, N. N. , Prescott, Beverly A., and Lester, L. P., (1981), Further Approx¬ imation to the Distributions of Some Transformations to the Sample Correla¬ tion Coefficient. Computer Science and Statistics: 13th Symposium on the Interface, Dept, of Stat. and Comp. Sc., Carnegie Mellon University, Pittsburgh, Pa. Wolde-Tsadik, Girma, (1980), Sample Sizes in the Interval Estimation of the Correlation Coefficient, The American Statistician, 34, 227-28. ANNOUNCING MOUNTAIN LAKE BIOLOGICAL STATION SUMMER COURSES 1983 First Term (June 13 - July 16) Biology of Birds David W. Johnston, George Mason University Invertebrate Life: Pattern and Diversity James L. Gooch, Juniata College Population Biology of Plants Janis AntonovJ.cs, Duke University Leslie Real, North Carolina State University Taxonomy of Vascular Plants James W. Hardin, North Carolina State University Techniques & Materials for Scientific Illustration Esta L. Johnston George Mason University Second Term (July 18 - August 20) Animal Behavior Steven P. Martindale, University of Virginia Ecological Genetics David A. West, Virginia Polytechnic Institute & State University Field Botany of the Southern Appalachians Charles R. Werth, Indiana University Insect Ecology Earl D. McCoy, University of South Florida Vertebrates of the Southern Appalachians Arthur J. Bulger, University of Virginia Joseph Travis, Florida State University Jerry 0. Wolff, University of Virginia Workshop in Alio zyme Techniques Charles R. Werth, Indiana University Scholarships Available: Service Awards for students covering room and board costs. Post-doctoral Research Awards (10 weeks) For further information and application write to: Dr. J. J. Murray, Jr., Director Mountain Lake Biological Station Gilmer Hall University of Virginia Charlottesville, VA 22901 VIRGINIA JOURNAL OF SCIENCE OFFICIAL PUBLICATION OF THE VIRGINIA ACADEMY OF SCIENCE THE VIRGINIA JOURNAL OF SCIENCE EDITOR: Stewart Ware Dept, of Biology College of William and Mary Williamsburg, Va. 23185 BUSINESS MANAGER: N. J. Fashing Dept, of Biology College of William and Mary Williamsburg, Va. 23185 ©Copyright, 1983 by the Virginia Academy of Science, The Virginia Journal of Science (ISSN 0Q42-658X) is published four times a year (Spring, Summer, Fall, Winter) by the Virginia Academy of Science, Department of Biology, University of Richmond, Richmond, Va. 23173. Second Class Postage paid at Richmond, Virginia and additional entry at Harrisonburg, Virginia 22801. The Virginia Academy of Science and the Editors of the Virginia Journal of Science assume no responsibility for statements or opinions advanced by contributors. Subscription rates for 1983: $27.50 per year, U.S.A.; $35.00 per year, other countries. All foreign remittances must be made at par U.S. dollars or their foreign equivalent. Back issues are available for $7.50 per issue postpaid. Changes of address, including both new and old zip codes, should be send promptly upon moving to the following address. POSTMASTER: Send address changes to Blanton M. Bruner, Executive Secretary- Treasurer, Virginia Academy of Science, Department of Biology, University of Richmond, Richmond, Va. 23173. All correspondence relating to remittances, advertising (rates will be sent upon request), subscriptions, missing issues, and other business affairs should be addressed to Business Manager. Instructions to Authors All manuscripts and correspondence about them should be addressed to the Editor. The Virginia Journal of Science welcomes for consideration original articles and short notes in the various disciplines of engineering and science. Cross-discriplinary papers dealing with advancements in science and technology and the impact of these on man and society are particularly welcome. Submission of an article implies that the article has not been published elsewhere while under consideration by the Journal. The original and two copies of each manuscript and of all figures therein are required. Authors should submit names of three potential reviewers. All manuscripts must be double-spaced through¬ out. The title, author’s name, affiliation and address should be placed on a covering page. An abstract (on a separate sheet) summarizing the text, particularly the results and conclusions, is required. After revision and final acceptance of an article, the author will be asked to furnish an error-free, camera-ready copy of the manuscript typewritten single-spaced with a black carbon ribbon on white bond paper. The text should follow the general format used by professional journals in the author’s disci¬ pline. Literature cited in the text should follow the name-and-year format: Fujishima and Honda (1972), or Spry ( 1969), or Guliday (1971). On the Literature Cited pageattheend of the article each reference should include author(s), year, title of article, title of journal (using standard abbrevia¬ tions), volume number, and first and last page of the article. For a book, include author(s), year title, pages or number of pages, and publisher and city of publication. Examples: Fujishima, A., and Honda, K. 1972. Electrochemical Photolysis of Water at a Semiconductor Electrode. Nature 238: 37-38. Spry, A. 1969. Metamorphic Textures. Pergamon Press, New York. 350 pp. Guliday, John E. 1971. Pleistocene History of the Appalachian Mammal Fauna. In Distributional History of the Southern Appalachians, Part III. Vertebrates (Perry C. Holt, ed.). pp. 223-262. VP1 & SU, Blacksburg, Va. Each figure and table should be mentioned specifically in the text, with all figure numbers and legends typed consecutively on separate sheet, and the figures identified by author’s name and figure number in pencil on the back. Table numbers and legends should be included as part of the table. Authors will be allowed 15 printed pages (including figures) free, but payment of $25 per page will be charged for the 16th and subsequent pages. VIRGINIA JOURNAL OF SCIENCE OFFICIAL PUBLICATION OF THE VIRGINIA ACADEMY OF SCIENCE Vol. 34 No. 2 Summer 1983 TABLE OF CONTENTS ARTICLES Virginia’s Eminent Deceased Chemists. Charles H. Fisher, Roanoke College. 37 Degree of Aflatoxin B1 Sensitivity in Virginia Natural Populations of Drosophila melanogaster . Sharon Delawder and Joseph P. Chinnici, Virginia Commonwealth University. 48 Callixylon Wood from the Greenland Gap Group (Upper Devonian) of Southwestern Virginia. Judith E. Skog, George Mason University. 58 Prevalence and Distribution of Lungworms of Swine Raised in Small Farm Units of Southside Virginia. Dilip K. Sen, Euzebio B. Damazio, and Peter J. McNerney, Virginia State University. 65 A Computer Data Acquisition and Analysis Program for the Labora¬ tory. Russell W. Vaughan and David A. Jenkins, V.P.I. & S.U. 70 Distribution of Pinus virginiana & P. taeda in the Peninsula of Virgin¬ ia. Cindy Larson Rice and Stewart Ware, College of William & Mary. 80 A Rapid and Convenient Procedure for the Synthesis of Radio-labeled Di- and Tri-peptides. Lloyd Wolfinbarger and John Castellano, Old Dominion University. 87 Errata Bell, Bovard, & Stratton 93 Announcement Research Grants 94 Actual date of publication: May 24, 1983 Virginia Journal of Science Volume 34, Number 2 Summer 1983 Virginia’s Eminent Deceased Chemists7 Charles H. Fisher Chemistry Department Roanoke College Salem, V A 24153 ABSTRACT The lives and achievements of sixty-five eminent deceased Virginia chemists are discussed. Thirty-four of the sixty-five were born or educated in Virginia, whereas the other thirty-one contributed to chemistry and chemical education while living in Virginia. Seventeen of the sixty-five died during the 19th Century, fifteen during 1901-1950, and thirty-three during 1951-1980. INTRODUCTION It is appropriate in any discussion of Virginia's eminent deceased chemists and chemical engineers to start with the reminder that Virginia was the home of the first permanent English settlement in the Western Hemisphere, which was esta¬ blished at Jamestown, Va., in 1607. In early colonial days, Virginia's vast territory included much of today's eastern United States. Hence Virginia has a longer history than its sister states, and at one time was much larger in square miles than any other state. Virginia's long history and early terri¬ torial size might be partly responsible for the fact that seven of the first twelve presidents of the United States were Virginians . Many of the presidents and other leaders from Virginia were knowledgeable or appreciative of science and engineering. Washington was a surveyor and engineer with a deep interest in scientific agriculture. Jefferson, a graduate of the College of William and Mary, had wide scientific interests. William Henry Harrison, the ninth president of the United States, attended Hampden-Sydney College and studied medicine. George Rogers Clark, a prominent Virginian, was a surveyor. Joseph R. Wilson, father of Woodrow Wilson, taught chemistry at Hampden- Sydney College. The climate was healthy for science in ^Based upon a talk given by the author before a meeting of the Division of History of the American Chemical Society in Atlanta, Ga. , March, 1981. 38 THE VIRGINIA JOURNAL OF SCIENCE Virginia very early in its history. It seems appropriate now to look back and to pay tribute, however brief, to Virginia's eminent chemists who lived and died during the approximately 370 years ‘since the Jamestown settlement. A Virginia chemist is herein defined as one who was born or educated in Virginia, or who contributed importantly to chemistry while living in Virginia. Deceased Virginia chemists were considered eminent if they were included in "American Chemists and Chemical Engineers," (Miles, 1976); designated "distinguished" in American Men of Science, 5th Ed. , 1933; awarded important prizes and honors; and/or noted for outstanding performance as educators, researchers, inventors, authors, administrators, or business executives. The challenge of presenting effectively information on sixty-five eminent deceased chemists is formidable. To facil¬ itate the operation, the following plan was developed: First, the individuals were categorized by periods: (1) Those individuals deceased during the 19th Century, or before the year 1901; (17) (2) Those deceased during the period 1901-1950; (15) (3) Those deceased during the period 1951-1980; (33) Within these periods, the individuals were categorized as those who were (1) born in Virginia, (2) educated in Virginia, or (3) who lived in Virginia during a considerable part of their careers . EMINENT DECEASED VIRGINIA CHEMISTS IN THE 19th CENTURY Although there seems to be no record of eminent Virginia chemists in the 18th century, there were a considerable number in the 19th century. Wyndham D. Miles's book entitled, "American Chemists & Chemical Engineers," (American Chemical Society, 1976) describes the careers of 17 eminent Virginia chemists who lived and died in the 19th century (Table 1) . Several of these were particu¬ larly outstanding: J. P. Cushing, President, Hampden-Sydney College, 1821-35; H. Draper, M.D., recipient. Congressional Medal of Honor; J. W. Draper, M.D., President, Medical School, New York University, 1850-73; recipient, Rumford Medal, 1875; and first President of the American Chemical Society; R. M. Patterson, M.D., Director, Philadelphia Mint, 1835-51; W. B. Rogers, Founder and first President, Massachusetts Institute of Technology, 1865-82; J. L. Smith, President, American Chemical Society, 1877. Four of the 17 eminent Va. chemists of the 19th century were born in Virginia's small towns and rural areas; four were born in Philadelphia; and five were born in Europe. Ten of the 17 had M.D. degrees (Table 1). P.K. Rogers, R.E. Rogers^ Mitchell, and Patterson got their M.D.'s from the University of Pennsylvania; J. C. Draper and H. Draper from New York University; Smith from the University of South Carolina; J.B. Rogers from the University of Maryland; and J. W. Draper from Jefferson Medical College. De Chalmot had a Ph.D. from Gottingen. Four of the 17 (Patterson, 1828-35; W. B. Rogers, Table 1. Eminent Chemists Born, Educated or "Careering" in Virginia in the 19th Century a, b, c. Those with M.D. degrees are marked with an asterisk. EMINENT VIRGINIA CHEMISTS 39 g 0 0 5-1 0 U 0 > . > > •g -H X g g g D D 2 o 0 cm x X 2 X g g o 0 0 • >H >H M-i rH 2 •H 3= CD 0 0 0 2 CO 2 2 0 • > -I • >i D 0 . g 2 - C • • 0 >1 0 0 0 2 u cm > > fd • 2 • • • X > > > 0 • 2 2 -H id s g g g 5 s d o D • t t ( u • >i >i >i fd >i >i Q) fd g g g > co co x > fd fd fd > a cm x £ . . • -h g £ w £ ggggtdfdfdo i — 1 > > i — ! rH >i •H •H >H >1 0 g g 0 0 g D 2 g g g g g ggd g g o 0 >i X X >i >i >1 >1 0 0 0 -P 0 0 0 •H cm g g g g g g g > cm cm g 2 -p cn X 0 0 0 0 0 0 0 2 o 0 g 0 • 2 >H >i 2 2 2 2 • n. • g • £ • u -h o > > > o > -p 4-i g -p g •H • 5 . ■rH -H in .g g 2 2 X 2 g £ 0 0 £ £ £ £ ' g g x g 0 0 g «o w D 2 2 £ S D 2 o D a 2 PM o 0 0 > 0 > • > 0 • > 0 V 6 . u • u u - 0 • . • . g 0 U 2 TJ CO 2 2 5 cn g 2 2 2 0 g cn 0 - x X 0 g 0 0 0 - g 0 *■ x 0 0 g r0 CM PM PM 0 0 pm g 0 g 2 u £1 w g -p 0 g •H g g V •» *. 0 0 0 0 0 ' M 0 0 0 cq 0 0 •rH 0 . . • £ 0 g g g • 0 g £ g a o i — 1 o 0 0 0 2 i—i 000000 2 0 cm g X g i — I i — 1 2 -P g 1— 1 r—i 1 — 1 1 — 1 x 1— 1 -p 1 — 1 0 •H o •pH •H 2 •rH r- 1 0 Cn 0 Cn 2 0 1— 1 . — 1 x g 0 g x! x X 0 X cggxog0O CO CM 2 CM PM PM PM CQ u HHWCMPiWCQK * He — * * — O'! OO 2 1 i m ^ 00 , — „ , — . r— 00 1 1 CO CO CN 1 CD 2 ro 00 r- ro cn 00 1 CO 0 1— 1 I— 1 i — 1 1—1 1 l—l 1 0 g rH 0 2 0 rH x 1 PQ g £ 0 g 2 +J g rH Td rH x 2 ~ — g 1 — 1 0 r— 1 0 W g 0 • g g 0 rH g rH x X! — ' o X E a 0 pm x g 0 a -h 0 -g g g 2 u g 0 -P 0 g 0 0 a 4-> s x: g ■H X! g >1 rd 0 2 >i i g PI 0 0 -g 2 0 0 00 CQ 0 g g g w (D r—l g 0 u 0 g pi g g -g 2 £ x g £ 2 g g £ +J - 0 X Pi - > 2 0 0 0 -H 0 0 X >i 0 0 g - 2 0 -p cq - w 2 to CQ 2 5 2 pi 0 PQ x cm 0 g 2-0 i— i 2 2 x 0 - X E i— i - «. - - V v *. - tji 0 Cn »■ 0 1 — 1 0 g g g 0 0 0 0 ^0ggggrHd3 X J •H 0 0 g g g g 0 g -g 0 -g 0x2 o 2 CM Cm 0 0 0 0 -P 002XX ago -p 2 0 0 Cn tji i Cn i Cr * -H 0 tJI rH x 0 0 u •H g g g 0 0 0 0 £ oo-H0gg00 2 Pi a a Pi Pi Pi Pi CO 2Pi2CnUa2TS ID r- • • cn 3= X g 0 — 0 -g 0 > 0 g 0 cn 0 g 2 0 tT1 2 g g 00 •H 0 -g 2 0 0 S X CO 0 •• rH v X . 1 — 1 ^ 2 £ 0 ", 0 0 2 r- X cn| X u 0 0 X • 0 . X 2 £ £ 0 < £ co X 0 g - g • 0 ID 2 2 CM 0 cn 2 • 1 — l 0 0 0 g > £ E 2 0 - 0 - 0 0 X 0 g X 0 X 0 0 0^ cn 0 1 — 1 g g 0 1 0 ■g X o> X Cn 0 X g >1 *D rH w 0 1 — 1 0 £ 2 0* . 0 0 • 0 > 0 x 0 2 2 •g 0 > g g £ •H 2 D 0 £ g cn X 0 2 u X u X 2 g 0 0 > 0 1-3 0 X 0 0 g 0 0 g 2 ^ X 0 0 0 0 x 2 x 2 •g x 0 g £ CO 0 X 0 2 a, 0 X 0 2 0 U 0 g 0 0 0 0 g £ 2 0 CM 0 0 0 S 0 0 0 >H g •H X g a g X PM 0 0 g 0 • £ 0 0 g 5 < X X 0 • = 0 £< 0 2 g - 2 - 0 0 • 0 • 0 2 g 0 rH x 2 x . •H 0 g 2 2 £ *■ 0 g x CM • 0 g 0 a 2 0 x g . g g 0 0 2 eh 0 X 0 £ £ 0 • 1 — l 0 0 g w 0 g 0 • X 2 Cm 2 5 rt 0 i—* 3- P „ _ „ G 0 0 0 3- P- p G 3 0 O O O 3 3- g 00 3 3 G 0 G 0 G 0 rt G 0 0 CD CD P CD O Cl G 00 n G G 31 P 00 3 0 3- 0 Pj CD CD ft '3 3 U1 N) G 00 0 00 0 P ft 3 Pj Pj 3- 3- i— 1 1 G NO 0 G Cl 3 G p 3 0 O 3- 3- 0 O 0 00 G G 00 1 3 00 G M 3 1 G P £ 3 3 3 P 3 -J VO 00 U> G 0 -J 00 1 k; G 00 vQ G 3 00 U) Cl M VO P O G VO G G s; 01 01 Hi O 1 O 4^ 1 O 0 1 O VO 4^ 4^ 00 00 G ' " 1-5 0(1) ro s: g £ s: 3- 0 p p 3- D 0 (0 H H iQ 3 It H 01 G 0 G 0 O P ' (D ft 3 3 - - - - C_| cr c_, c-i c_i 0 0 0 * tsi td 0 P 3- 01 01 G * * > SO D 1-5 0 p cr ro 3 C 01 3 G 0 O (D 0 3 G ^ 3 3 < G vQ vQ 3-0 O r+ > G ' 3 O P 3 c-i S £ D < O 0 Pi P 0 3 pi vQ G 3 0 CL 3 3 Pi 01 (D 3 (D G ' ' pj fc< G CD - CD S 50 1-5 • 3- •» O O 3* 31 G I 4^ G G | CT) U1 00 G 0 G 3 V£> |0 G uo VO G O 00 G oo vo un 0 G G 3 0 0 4* UO G |Hi G N) VO on | 0 vo l 1 Ci 0 KD >X) 3 0 Ul G N> 00 K) ICG -• G G G 1 P 4^ G 3- 3 ^ Cl ' — 00 ^ ui 1 |0 O VO VO G - Cl • 0 0 1 tr — .is 00 |G— G ' — 4Jj 4^ VO 1 1 ft is 1 IS VO G"J ui LO 4^ s; 0 3- G 4^> P- un • P VO | o 1 0^ 0 U> 1 — IS £ ft 00 |G K 31 31 3- O 0 • • Pj P* 0 IG P k: 0 3 U) 4^ 0 3 0 0 rt 3- 3 n ro g H cn G D G CEJ td H O D> ffi S * • rt 0 0 P 0 3 rt 0 0 0 P P 3 G G P n i_ j • 0 0 p 3 3 G X 0 0 Hi 0 £ G G 0 P G 3 i 3 Hi >• G 0 P rt 3- G P Hi 3 3- rt ct P 3 0 t 0 3 3 s: 3- 3 P cr 0 0 3- 3- G 3 Pj fl> G M 0 P 3 G P 0 3 W G 3 3 O 0 P 0 3 3 P G 0 ft Pj p 0 3 P 0 0 P rt 3 P C) 0 • G 0 G 3 0 < 0 0 3 3 3 ft ft G 1 31 3- rt ' 0 p 3- P 0 G 0 0 0 0 0 cn QJ P P P > <« G G ' ' - 0 3: a 3 0 rt < G O ■» < O Pj P 0 0 ft £ * 0 3- S £ £ < H- 0 P rt 3 0 • Pj •« ft £ • Pj Pj P G • 0 8 3- 0 • cn < • • • G - k 0 - o • P 0 •» P - G • ■* < '■ p < s3 G 0 < • p 3 P • 3 'J • 3- Cl P Q - GOG Q G G p G G < G gr 6? G 0 t? 0 G 0 01 P D| 3 P 3) P § S P ft 3 G P P* rt 3- 3-13 3- 3- HI 3- 3- 0 • P 3 P ft •o < < * < < < • • < P n p* 0 G 0 3- • * cn • e> • in in • G ffi K P G < < Pj < < cn ^ ‘S.’S. < P 0 0 0 0 P P * P P G p P 3- 3 33 33 P • * ^ » • * G G P- P- < < < < P (D 3 G n G G ffi Pj G D Pj G n < n w G 0 P 0 P P P P • P P P 0 P 0 3 • 0 3- 3- 3- 3 3) 2 < 3J 3- 3 • 1 o 2 o 0 < p < < tl 0 • 3- 0 < 3 3 3 • P ■* ’ P Pj P • • H • K 3 rt n 0 rt P o 0 G SO 0 < < cn p 0 < 0 Pj 0 G p 0 VO 3- G P P 3 P P Hi P G 0 3 3 O . p. 0 • 0 P 0 00 G < G o < ft < 3 G • 3 P 3- 0 P 3 p k: 3- 0 P G • • P P P • P 0, Table 2. Eminent Chemists Born, Educated, or "Careering" in Virginia and Deceased 1901-1950— An asterisk means the chemist was called "distinguished" by "American Men of Science"1933 EMINENT VIRGINIA CHEMISTS 43 Virginia: Baskerville, Chairman of the Chemistry Department, University of North Carolina, 1900-04; and Bingham, Head of the Department of Chemistry & Metallurgy, Lafayette College, 1916- 39. Two of the 15 became college presidents: Dabney, President of both the University of Tennessee and the University of Cin¬ cinnati, 1904-20; and Venable, President of the University of North Carolina, 1900-14. Four of the 15 were chief officers of state scientific societies: Baskerville, President, North Carolina Academy of Science; Jones, President, Virginia Academy of Science; Venable, Founder and first President, Elisha Mitchell Scienti¬ fic Society, 1833; and Zeisberg, Chairman, Delaware Section, American Chemical Society. Seven of the 15 occupied high positions in national or international scientific societies: Baskerville, Vice-President, American Association for the Advancement of Science, 1903; Dunnington, Secretary, Chemistry Section, Am. Assoc. Adv. Science; Mallet, President, American Chemical Society, 1882; Meade, Vice- President, American Institute of Chemical Engineers; Reese, Vice- President, International Union of Pure and Applied Chemistry; President, Manufacturing Chemists' Association, 1920-23; Presi¬ dent, American Institute of Chemical Engineers, 1923-25; and President, American Chemical Society, 1934; Venable, President, American Chemical Society, 1905; and Zeisberg, President, American Institute of Chemical Engineers, 1938. Four of the 15 advanced to high positions in industry: Magruder, Chief Chemist , F. S. Royster Guano Co., 1915-45; Meade, Chief Chemist, Edison Portland Cement Co., and General Manager, Tidewater Portland Cement Co.; Reese, Chief Chemist, New Jersey Zinc Co., and Chemist & Manager, E.I. duPont de Nemours & Co., 1902-31; and Zeisberg, Chemist & Manager, E.I. duPont de Nemours & Co. Seven of the 15 held important positions in State organiza¬ tions: Baskerville, Chemist, North Carolina Geological Survey; Dabney, Director, North Carolina Experiment Station; Jones, Chemist, Virginia Department of Agriculture, 1913-31, and Chief Chemist, Commonwealth of Virginia, 1931-44; Kastle, Director Kentucky Agricultural Experiment Station, and Dean, College of Agriculture, 1912-16; Magruder, Chief Chemist, Virginia Department of Agriculture, 1900-15; Mallet, Superintendent, Confederate States Ordinance Laboratory, 1862, and Weems, Chief Chemist, Virginia Department of Agriculture, 1915-30. Three held positions in the following Federal establish¬ ments: Bingham, Physicist, U.S. Bureau of Standards, 1916; Dabney, Assistant Secretary, U.S. Department of Agriculture; and Kastle, Chief Chemist, U.S. Hygienic Laboratory, 1905-09. EMINENT VIRGINIA CHEMISTS DECEASED 1951-80 A total of thirty-three eminent Virginia chemists are listed (Table 3) as having died during the period 1951-80. Eleven of these were born in Virginia and four others received at least some of their higher education in Virginia (Table 3) . Eighteen others are categorized as Virginia chemists because substantial portions of their careers were spent in Virginia. Most of the thirty-three chemists listed in Table 3 received 44 THE VIRGINIA JOURNAL OF SCIENCE Table 3. Eminent Chemists Born, Educated or Careering in Virginia and Deceased 19 51-19802. By Birth Career Allison, Fred (1882-1974)- Cox, Edwin (1902-1977)2. Gwathmey, Allan Talbott (1903-1963)— Jones, J. Claggett (1907-1965) Potter, Thomas Willoughby, Jr. (1902-1972) Reid, E. Emmet (1872-1973)®. Sherman, Henry Clapp (1875-1955)— Smith, Foley Foster (1905-1969) ^ Trout, William E. , Jr. (1903-1979)- Veazey, William Reed (1883-1958) f Watson, John Wilbur (1888-1962) d Glade Spring, Va. Richmond, Va. Richmond, Va. Roanoke, Va. Floyd Co. , Va. Fincastle, Va. Ash Grove, Va. Danville, Va. Clifton Forge, Va. Chase City, Va. Chatham, Va. U.Va. , Emory Henry VMI VMI, U.Va. VPI & SU Roanoke College U. Richmond, John Hopk. Columbia U. Va. Johns Hopk. Johns Hopk. U.Va. Auburn V. Carol. Ch< U.Va. Comm, of Va. JBSC Johns Hopk. Columbia Comm, of Va. U. Richmond Case School VPI & SU By Education Bull, Fred Warren (1912-1979)— Desha, Lucius Junius (1883-1967)— Lind, Samuel Colville (1879-1965)— Sherrill, Mary Lura (1888-1968) f By Career Benton, Arthur Ferguson (1895-1979)— Chase, Harold Mason (1872-1960) Detwiler, Samuel Bertolet,.-Jr.(1909-1978) — Edgar, Graham (1887-1955)1 German, Leslie (1909-1980)- Gilreath, Esmark Senn (1904-1979)— Guy, William George (1899-1969) Hitchcock, Lauren Blakely (1900-1972) — "Howe, James Lewis (1859-1955) f Kupchan, S. Morris (19 22-19 76)2. Lutz, Robert Elliott (1900-1976)— Major, Randolph Thomas (1901-1976)— Robey, Ashley (1903-1965)^3. Scherer, Philip Carl (1893-1960)- Scribner, Allison Kenneth (1898-1976) Small, Lyndon Frederick (1897-1957)— Vilbrandt, Frank Carl (1893-1960)^ Yoe, John Howe (189 2-19 78)2.'®. Erwin, Tn. VPI & SU VPI & SU Cynthiana, Ky . Wash. Lee Wash. Lee McMinnville, Tn. Wash. Lee U. Minnes. Salisbury, N.C. Randolph Macon Mt. Holyoke Cranford, N.J. Princeton U. Va. Lowell, Mass. M.I.T. Dan River Inc. Wabasha, Minn. Geo. Wash. USDA Fayetteville, Ark. Yale . U. Va. Dayton, Ky . U. Cincin. VMI N. Wilkesboro, N.C. U. No. Carolina Wash. Lee Carbonear, Newfoun. U. Chicago Wm. Mary Paris, France M.I.T. U.Va. Newburyport, Mass. Gottingen Wash. Lee New York, N.Y. Columbia U.Va. Boston, Mass. Harvard U.Va. Columbus, Oh. Princeton U.Va. Fort Worth, Tx. Cornell Roanoke Col. Brooklyn, N.Y. Brown VPI & SU Boston, Mass. Harvard Va. Chemicals Allston, Mass. Harvard U.Va. Fostoria, Oh. Ohio State VPI & SU Oxford, Ala. Princeton U.Va. l two or more states; an asterisk means the chemist was called "distinguished" by American Men of Science, 1933; JBSC is Journal Box Servicing Corp. b. American Men & Women of Science, 1977. c. Who’s Who in America 1975. d. American Men of Science, 1949. e. World's Who's Who in Science, 1968. f_. W. D. Miles, "American Chemists & Chemical Engineers", 1976 , Am. Chem. Soc. g. Chemical Who's Who, 1951. earned Ph.D. degrees. Some of the schools that awarded the degrees are the University of Virginia (4) , Harvard (3), Johns Hopkins (3) , and Princeton (3) . Most of the 33 chemists achieved eminence in academic institutions. Three chemists (Cox, Potter, and Scribner) were unusually successful in industrial careers. Six chemists (Reid, Sherman, Veazey, Lind, Hitchcock and Small) had careers both in academia and industry or in academia and government. Detwiler and Jones pursued careers in Federal and State government, respectively. Smith worked both for industry and the Common¬ wealth of Virginia. Several of the 1951-80 deceased chemists were elected to high offices in national or state technical societies: S.C. Lind, President, American Chemical Society, 1940; R. T. Major, Board of Directors, American Chemical Society, 1955-60; W. R. Veazey, President, Electrochemical Society, 1946; Leslie German, Chairman, Virginia Blue Ridge Section, Am. Chem. Soc., 1950; A. T . Gwathmey, President, Virginia Academy of Science, 1954; and F. F. Smith, Chairman, Virginia Section, Am. Chem. Soc., 1946, EMINENT VIRGINIA CHEMISTS 45 and President, Va Academy of Science. The following reached high positions in academia: Fred Allison, Dean, Graduate School, Alabama Polytechnic Institute, and Director, Auburn Research Foundation, 1949; F. W. Bull, Department Head, VPI & SU, 1965-78; Dean, Graduate School, VPI & SU, 1965-78; L. J. Desha, Dean, Washington & Lee, 1946; S. C. Lind, Chief Chemist, U.S. Bureau of Mines, 1923-25; Dean, Insti- tue of Technology, Univ. of Minnesota, 1935-47; A.F. Benton, Department Chairman, University of Virginia, 1928; Leslie German, Department Head, Va. Military Institute, 1942; E. S. Gilreath, Department Head, Washington & Lee, 1946-70; W. G. Guy, Depart¬ ment Head, College of William & Mary, 1946-68; R. E. Lutz, Department Head, U. of Va. , 1951-53; Ashley Robey, Department Head, Roanoke College, 1941-65; Mary L. Sherrill, Department Head, Mt. Holyoke, 1946-54; W. R. Veazey, Department Head, Case School of Applied Sciences; Board of Trustees, Westminister College; Director, Dow Chemical Co., Vice President, Dowell, Inc., Director, Dow Corning Corp; F.C. Vilbrandt, Department Head, U. of No. Carolina, 1927-30; Department Head, VPI & SU, 1935; J. W. Watson, Department Head, VPI & SU, 1925; and J. H. Yoe , Department Head, U. of Va. , 1953-57. These reached high positions in industry: H. M. Chase, Director of Research, Dan River, Inc. ; Edwin Cox, Vice Presi¬ dent, Va. Carolina Chemical Corp., 1949-57; President, Tobacco Byproducts and Chemical Corp; Graham Edgar, Research Director, Ethyl Corp., 1924-32; Vice President, Ethyl Corp., 1932; Vice President, Ethyl Dow Chemical Co., 1934; L. B. Hitchcock, President, Los Angeles Air Pollution Foundation, 1954-57; R. T. Major, Vice President and Scientific Director, Merck & Co. , 1947-53; T. W. Potter, Vice President, Journal Box Servicing Corp., 1940-68; A. K. Scribner, Vice President, 1950, President, 1957, Va. Chemicals; Director, Norfolk, Franklin & Danville Railroad. The following were important in State and Federal government: S. B. Detwiler, Jr., Technical Program Specialist, Agricultural Research Service, U.S. Department of Agriculture, 1953-72; H. C. Sherman, Chief, Bureau of Human Nutrition, U.S. Department of Agriculture; L. F. Small, Research Specialist, National Insti¬ tutes of Health, and L. F. Small, Director, Drug Addiction Laboratory, U. of Va. From the foregoing, it is clear Virginians have reason to be proud of its chemists beginning in its early history and continuing today. The large volume and high quality of current chemical research in Virginia give assurance there will be ample justification for pride in the future. ACKNOWLEDGMENT Appreciation is expressed for valuable assistance received from: Stewart Ware, C. W. Bondurant, Jr. , L. C. Bostian, O.C. Cleveland, R. D. Cool, Lois Carlin Fisher, L. Aubrey Goodson, Jr., Paul D. Henson, G. W. Irving, Jr., Henry A. McGee, Jr., Marshall Mead, J. W. Midyette, Jr., Mrs. T. W. (Jean) Potter, Raymond B. Seymour, Jesse Thompson, Gene Wise, and J. H. Wise. 46 THE VIRGINIA JOURNAL OF SCIENCE LITERATURE CITED Bohning, J. J. , 1982, Prestige vs. Practicality in Selecting the First President of the ACS, Chemical & Engineering News, Mar. 8, pp. 31-34. Brown, C.A. , 1943, Thomas Jefferson's Relation to Chemistry, J. Chem. Ed. 20_, 574, Browne, C.A., and Weeks, M.E., 1952, History of the American Chemical Society, American Chemical Society, Washington, D.C Catell, Jaques, Editor, 1933, "American Men of Science," 5th Ed. Catell, Jaques, Editor, 1949, "American Men of Science," 8th Ed. The Science Press, Lancaster, Pa. Jacques Catell Press, 1971-73, American Men & Women of Science, 12th Ed. Jaques Cattell Press, 1977, R. R. Bowker Co., N.Y., American Men & Women of Science, Chemistry. Cook, R. C. , Editor, 1966-67, Leaders in American Science, Vol. VII, Who's Who in American Education, Inc., Hattiesburg Ms. 39401. Debus, A.G., Editor, World's Who's Who in Science, 1968, A.N. Marquis Co., Chicago, Ill. Gillispie, C. C. , Editor, Dictionary of Scientific Biography, Charles Scribner's Sons, New York, 1972. Guy, W. G. , 1950, Early Chemistry, The James River Basin, Acad. Sci., Richmond, pp. 565, 570-71, 575. Haynes, William & Downs, W. D., 1951, Chemical Who's Who, Third Edition, Lewis Historical Publishing Co., Inc., New York. Irving, G. W. , Jr., 1979, Samuel Bertolet Detwiler, Cosmos Club Vignette No. 178. Marquis Who's Who, Chicago, Ill., 1974-75, Who's Who in America, 38th Ed. Miles, Wyndham, D. , 1976, "American Chemists and Chemical Engineers," American Chemical Society, Washington, D.C. Mulder, J.M. , 1978, Woodrow Wilson, The Years of Preparation, Princeton University Press. Sabine, D.B., 1978, John William Draper, Chemistry 51, (4): 12-14. Skolnick, H. , and Reese, K. M. , 1976, A Century of Chemistry, American Chemical Society, Washington, D.C. EMINENT VIRGINIA CHEMISTS 47 Thompson, A.R., 1948, Harold Mason Chase: 1948 Olney Medalist, Am. Dyestuff Reporter, Oct. 18, pp. P684-P685. Trout, W. E. , Jr., 1976, The Physical Sciences in Virginia, 1609-1900 , Va, J. Sci., 27_(4): 181-190. Virginia Journal of Science Volume 34, Number 2 Summer 1983 AM. Degree of Aflatoxin JJi Sensitivity in Virginia Natural Populations 3 Sharon Delawder and Joseph P.|chinnici Department of Biology Virginia Commonwealth University Richmond, VA 23284 ABSTRACT Natural populations of Drosophila melanogaster were sampled from counties in Virginia reporting aflatoxin contamination of corn ^ an# crops and from Virginia counties reporting no aflatoxin contamination, Fggs from each population sample were placed on media containing 0.0, 0.5, and 1.0 ppm aflatoxin B-^ (AFB^). Data on egg-pupal and egg-adult develop¬ ment times, egg-pupal and egg-adult viability, and the sex and body lengths of adults were collected to determine the relative degrees of resis¬ tance of these samples to AFB-^ induced toxicity. Flies sensitive to AFB-^ show decreased viability, smaller adult body sizes, and significant elon¬ gation of development times. There appears to be significant variation in resistance to AFB^ among the populations correlating with the areas from which the flies were collected. In general, flies from aflatoxin contaminated areas are more resistant to AFB^ toxicity than flies from aflatoxin free ar¬ eas. Natural selection may be responsible for this correlation. Chromosome substitution analysis indicated that factors on chromosomes two and three provide increased resistance to AFBj, while X-linked factors enhance sen¬ sitivity . INTRODUCTION Aflatoxins are a group of secondary metabolites produced by certain strains of the fungi Aspergillus flavus and _A. parasiticus. Chemically, the aflatoxins are highly oxytenated heterocyclic compounds that contain the bisfuran moiety, either as the dihydrofuran system, as in aflatoxins B-^ and (AFB^, AFG-^) or as the tetrahydrofuran system, as in AFB2 and AFG2 [Ciegler and Bennett, 1980]. The prolific _A . flavus is ubiquitous in the soil and air in many parts of the world [Detroy et al. , 1971; Raper and Fennell, 1973]. Not all strains of A_. flavus are capable of toxin production; however, reliable evi¬ dence indicates that a major proportion of isolates encountered are aflatoxin producers,, with the most common toxin produced being AFB^ [Stoloff, 1977]. Although A., flavus is usually classified as a storage mold of agricultural products, it also can contaminate crops in the field or or¬ chard, particularly in the warmer, more humid growing regions of the AFLATOXIN SENSITIVITY IN DROSOPHILA 49 world [Anderson et al. , 1975; Hesseltine et al. , 1976], Scrr.e of the ag¬ ricultural products known to be contaminated with aflatoxin in significant amounts and incidence include peanuts, corn, grain sorghum, millet, cottonseed, copra, and figs [Stoloff, 1977], Plant stress caused by drought appears to be a contributing factor to the invasion of corn and peanut crops by A_. flavus and the subsequent production of aflatoxin [Dickens, 1977; Lillehoj and Hesseltine, 1977]. A drought-wet sequence in climatic conditions in 1977 was believed to con¬ tribute to aflatoxin contamination of corn crops in certain areas of Virginia [Llewellyn and Katzen, 1981]. Following another severe drought in Virginia during the 1980 growing season, abnormally high levels of aflatoxin were reported in corn crops, especially in the southeastern portion of the state; and a large percentage of the peanuts harvested in drought-stricken areas of Virginia also was contaminated with aflatoxin. Well distributed and sea¬ sonally abundant rainfall and moderate temperatures were believed to be factors in the low infestation of peanut kernels and shells in three peanut growing states in 1964 [Diener et al. , 1965]. Attack of plants by insects, which often increases during periods of drought [Lillehoj and Hesseltine, 1977], may also contribute to aflatoxin contamination. A 1977 survey of corn samples from North Carolina showed that, in general, the higher the percentage of insect-damaged corn, the higher the level of aflatoxin [Hesseltine et al. , 1981]. Anderson et al. [1- 975] observed insect damage in 90% of the corn samples that were contam¬ inated with aflatoxin. Aflatoxin (AFBj) is highly mutagenic in a wide variety of orga¬ nisms [Ong, 1975; Wong and Hsieh, 1976] and highly carcinogenic among vertebrates of many classes [Newberne and Butler, 1969; Patterson, 1973; Wogan, 1977]. Among invertebrates, especially insects, dietary AFB^ has insecticidal, larvicidal, and chemosterilizing properties [Lalor et al. , 1976]. Various wild-type strains of the fruit fly Drosophila melanogaster dif¬ fer in their responses to AFB-^ placed in the culture medium in which the larvae develop from the egg stage. Toxicity is demonstrated by reduced larval and pupal viability, a reduction in the length of the pupal cases and the adult flies, and an increase in the egg-to-pupal and egg-to-adult de¬ velopment times [Lalor et al. , 1976; Chinnici et al. , 1976]. On the basis of these criteria, several wild-type laboratory strains of D. melanogaster are quite sensitive, and others are relatively resistant [Llewellyn and Chinnici, 1978], Strains relatively resistant to AFB^ damage must be exposed to double or triple the dose given to a sensitive strain before similar levels of toxicity are expressed. The only detailed genetic analysis of the control of resistance to AFB^ toxicity has been performed using D. melanogaster. Chromosome substi¬ tution analysis of laboratory strains has shown that the toxic variation is due to autosomal gene differences [Chinnici, 1980]. The present study was undertaken to determine whether the unusually high levels of aflatoxin in the natural environment of parts of Virginia in 1980 had any effect on the AFB^-resistance levels of natural populations of D. melanogaster in these areas. Population samples taken both from counties with aflatoxin-contaminated corn and/or peanut crops and from counties without aflatoxin contamination were tested in the laboratory for degree of resis¬ tance to dietary AFB^. Chromosome substitution analysis was then used to investigate the genetic basis for variation in level of resistance to AFB^. 50 THE VIRGINIA JOURNAL OF SCIENCE MATERIALS AND METHODS Collecting Flies Samples of natural Drosophila melancgaster populations were collected in the Fall of 1980 from seven counties (25 samples) in Virginia reported by the Virginia Department of Agriculture to have high aflatoxin con¬ tamination of corn and/or peanut crops. Collections were also made in sev¬ en counties (15 samples) reported to have little or no aflatoxin contamina¬ tion. Table 1 lists and briefly describes all 40 sites. Quart size mason jars with a nylon net covering to keep out larger insects were used as traps to collect the flies. A banana and yeast mixture was placed in the bottom of the jars as bait. Traps were placed on the ground in locations where fruit flies were most likely to be exposed to A. flavus and/or aflatoxin (i.e. peanut warehouses; peanut, corn, soybean fields) and, where possible, orchards and dumpsters adjacent to corn or peanut fields. The traps were left in place for several hours after which the jars containing flies were sealed with a piece of cloth and transported to the laboratory. Flies were allowed to lay eggs in the traps and the par¬ ents and offspring were transferred to half-pint culture bottles containing standard culture medium. They were allowed to reproduce until a sufficient number of flies were available for experimentation. Media The culture medium used was a mixture of yeast, dextrose, and agar with several inorganic salts. Tegosept was added to inhibit mold growth. Aflatoxin (Grade A. Calbiochem, La Jolla, Calif.) dissolved in 10 ml of acetone was used to make a stock solution of 10 ppm AFB^. Equivalent amounts of acetone were added to the control medium (0.0 ppm AFB-^). The stock solution and control medium were used to make dilutions of 0.5 and 1.0 ppm AFB^. The control and AFB^ supplemented media were poured in¬ to 8 dram glass shell vials, stoppered with foam plugs, and refrigerated until used. Experimental Procedures 1. Testing for AFB^ sensitivity. Flies from each of the 40 samples were allowed to lay eggs for 12 hours in half-pint culture bottles contain¬ ing control medium. The eggs were then collected and groups of 25 were placed on small squares of blotting paper moistened with distilled water. Each square with 25 eggs was placed in a separate 8 dram glass shell vial containing medium supplemented with either 0.0, 0.5, or 1.0 ppm AFB^. The vials were incubated at 25 ± 1°C. Each treatment was replicated six times. As the cultures developed, data were collected daily on egg-pupal and egg-adult development times, egg-pupal and egg-adult viability, and the sex and body length of adults. 2. Chromosome substitution analysis. The results of the sensitivity tests were used to select a natural Drosophila population that was relative¬ ly insensitive (resistant) to the toxic effects of AFB-^ (*in Table 1) and a relatively sensitive population (**in Table 1). The chromosome substitution technique of Seiger, [1966], was used to mate these flies with Complete Multiple Inversion (CMI) stock flies generously provided by Dr. T. R. F. Wright of the University of Virginia. The CMI stock contains multiply in¬ verted X (FM6), second (SM5) and third (TM3) chromosomes, and nonin- verted second (Sp bw^) and third (Sb) chromosomes so that a CMI female has the balanced karyotype FM6/FM6; SM5/Spbw^; TM3/Sb. All second and AFLATOXIN SENSITIVITY IN DROSOPHILA 51 Table 1. Virginia counties (contaminated or uncontamined with _A. flavus and aflatoxin) and collecting sites, and the effect of larval growth in aflatoxin B-^ supplemented media on viability to the adult stage. Counties and Collecting Sites number of adults (x ± SEM)a _ 0.0 ppm*3 0.5 ppm 1.0 ppm Southampton (contaminated) 1. Apple orchard near peanut 22.33 + 0.51 19.17 + 1.09 13.00 + 1.51 and corn fields 2. Peanut warehouse trash can 19.50 + 0.39 18.33 ± 1.22 11.67 + 1.10 3. Pear tree near peanut and 20.17 + 0.76 12.33 ± 1.31 5.50 + 1.39 corn fields 4 . Peanut sheller shell heap 19.17 + 0.72 13.83 ± 1.07 13.67 + 1.12 5. In soybean field 19.83 + 0.44 11.83 ± 1.23 5.83 + 1.52 6. Nursery tomato plants 21.33 + 0.69 18.00 ± 0.67 12.67 + 1.12 7 . Peanut sheller near 16.83 + 0.72 11.00 ± 2.03 5.75 + 1.43 pokeberries 8. Apple trees near corn 21.83 + 1.21 19.00 ± 1.29 12.17 + 1.69 fields 9. Public dumpsters 9.50 + 0.99 9.17 + 0.93 8.33 + 0.31 10. In soybean field 23.00 + 0.33 16.00 + 0.62 8.17 + 0.86 11. In soybean field Prince George (contaminated) 21.33 + 0.39 18.33 + 0.84 5.17 + 0.80 12. Apple tree in corn field 22.50 + 0.87 15.50 + 0.74 9.67 + 1.52 13. Peanut and soybean field 22.00 + 0.47 18.83 + 1.04 8.50 + 1.12 14. Nursery near pumpkins 22.83 + 1.12 17.50 + 1.35 11.50 + 1.12 15 . Pear tree near peanut 20.17 + 0.90 17.80 + 1.34 5.17 + 0.80 and corn fields 16. Pear tree near peanut 22.67 + 0.73 17.67 + 1.07 9.33 + 0.90 field Isle of Wight (contaminated) 17. Pumpkin patch 20.17 + 1.40 15.33 + 1.02 11.00 + 0.94 18. In soybean field 21.83 + 0.49 10.67 + 0.77 5.17 + 0.76 19. In soybean field Surry (contaminated) 21.67 + 0.77 12.33 + 1.45 5.50 + 0.74 20. Public dumpsters 20.17 + 0.83 13.67 + 1.12 7.00 + 0.74 21. In soybean field 19.50 + 0.39 1.50 + 0.46 0 Sussex (contaminated) 22. In soybean field 23.17 + 0.90 17.00 + 0.47 8.83 + 1.23 23. Apple tree and soybean field 20.50 + 0.51 16.33 + 1.05 14.83 + 0.95 field near peanut processor* Nottoway (contaminated) 24. Apple and pear orchard 17.17 + 0.37 9.83 + 1.14 5.33 + 0.73 Franklin (contaminated) 25. Apple orchard 20.50 + 0.84 16.83 + 1.04 12.17 + 0.86 Chesterfield (uncontaminated) 26. Apple orchard 23.00 + 0.78 19.33 + 0.84 9.00 + 1.11 27 . Apple orchard 15.67 + 0.73 7.00 + 0.88 0.50 + 0.46 28. Apple tree 22.67 + 0.39 14.83 + 0.76 4.17 + 0.80 29. Apple orchard 20.50 + 1.41 13.83 + 1.16 5.50 + 0.94 52 THE VIRGINIA JOURNAL OF SCIENCE Table 1. (continued) Counties and number of adults (x ± SEM)a _ Collecting Sites 0.0 ppmb 0.5 ppm 1.0 ppm Rockingham (uncontaminated) 30. Apple orchard ** 31. Apple orchard 32. Apple orchard 33. Apple orchard Albermarle (uncontaminated) 34. Apple orchard 35. Apple orchard Roanoke (uncontaminated) 36. Apple trees 37 . Apple trees Augusta (uncontaminated) 38. Apple orchard Botetourt (uncontaminated) 39. Apple orchard Prince Edward (uncontaminated) 40. Pear orchard 15.67 + 0.31 8.33 + 0.45 1.67 + 0.39 16.83 + 0.98 5.00 + 1.05 1.67 + 0.39 10.83 + 0.72 6.33 + 1.02 2.33 + 0.39 18.17 + 1.19 17.00 + 1.39 15.17 + 1.30 18.67 + 0.90 15.67 + 1.48 7.00 + 1.20 22.17 + 0.28 16.33 + 1.28 10.17 + 1.32 19.50 + 0.99 6.67 + 0.61 0.83 + 0.44 19.67 + 0.81 12.50 + 1.31 7.17 + 1.36 20.00 + 0.41 9.67 + 0.90 2.33 + 0.56 15.17 + 0.80 9.00 + 1.05 4.50 + 0.51 14.50 + 1.31 14.83 + 1.35 13.67 + 2.18 a. Each treatment was replicated 6 times. Each vial initially contained 25 eggs. b. ppm AFB^ in the culture medium. *RX23, resistant strain used in the chromosome substitution analysis. **SX23, sensitive strain used in the chromosome substitution analysis. third chromosomes are recessive lethals. Mating schemes similar to those described previously [Chinnici 1971, 1980] were used to produce strains with known combinations of chromosomes from a resistant (RX23) and a sensitive (SX23) natural population. Figure 1 gives the chromosome consti¬ tutions of the substituted strains thus produced. 3. To test the hypothesis that presence or absence of aflatoxin in the environment has an effect on the sensitivity of natural populations to AFB^ induced toxicity, the 40 fruit fly populations were categorized into two groups: those collected from aflatoxin-contaminated areas and those collected from aflatoxin-free areas. Variables (adult viability, development times, etc) from each group were compared and tested for significant dif¬ ferences using analysis of variance. RESULTS Sensitivity levels of the natural populations Analysis of the data showed that for each level of AFB^ treatments (0.0, 0.5, and 1.0 ppm), considerable variation existed among the 40 nat¬ ural populations in egg-pupal and egg-adult viabilities, egg-pupal and egg-adult development times and body lengths of adult males and females. Data for egg-adult viabilities are given in Table 1 and graphed in Figure 2. AFLATOXIN SENSITIVITY IN DROSOPHILA 53 RX 2 3 | mm'm RXS23 | m — 3 RX2S3 | mm — 1 RX3S2 | \ k tttm 177777 vjTfn R23SX | H l ttttAW R3SX2 | [777/7 bzJL_J Vfffn R2SX3 QESaEEl SX23 FH E3FE] X 2 3 STRAINS CHROMOSOMES □ O.S AFB| H 1.0 AFB, i — — i — — T“ — r - r O 5 !0 IS 20 NUMBER OF ADULTS Fig. 1. Chromosome substitution study. Chromosome content of each substituted strain and the mean number of adults (± SEM) for 6 replications starting with 25 eggs each are given for 0.5 ppm AFB1 and 1.0 AFB1 treatments. Chromosomes derived from the resistant strain (RX23) are shown crosshatched, and those from the sensitive strain (SX23) as open rectangles. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 IS 16 17 18 19 20 21 22 23 24 25 U. o Fig. 2. Mean ± SEM values for six replications of 0.5 ppm AFB1 and 1.0 ppm AFB1 treatments of the 40 natural populations sampled. Collecting sites, numbered under the bars, are the same as in Table 1. A. The 25 population samples from Virginia counties contaminated with aflatoxin; B. The 15 population samples in Virginia counties not contaminated with aflatoxin; C. Combined values for the contaminated (C) and the uncontaminated (U) populations. 54 THE VIRGINIA JOURNAL OF SCIENCE Figure 2C illustrates that at each of the concentrations of AFB^ , overall egg-adult viability was significantly greater for populations from aflatoxin-contaminated areas (x ± SEM for 0.5 ppm = 14.71 ± 0.66, for 1.0 ppm = 8.64 ± 0.70) than for populations from uncontaminated areas (x ± SEM for 0.5 ppm = 11.70 ± 1.14, for 1.0 ppm = 5.71 ± 1.15; ANOVA con¬ taminated vs uncontaminated for 0.5 ppm: F=4 .41 , P=<.05 , and for 1.0 ppm: F=5 .02 , P=<.05). At 0.5 ppm AFB-^ , fruit fly populations from aflatoxin-contaminated areas had significantly shorter egg-adult develop¬ ment times than those from uncontaminated areas (ANOVA F=17 .24 , P=<.001). Fly populations from contaminated areas also had significantly greater egg-pupal viability (ANOVA for 0.5 ppm: F=52.44, P=<.001, and for 1.0 ppm: F=37 . 17 , P=<.001) and shorter egg-pupal development times (ANOVA for 0.5 ppm: F=21.27, P=<.001, and for 1.0 ppm: F=22.02, P= <.Q01) than fly populations from uncontaminated areas when tested at 0.5 and 1.0 ppm AFB^. No significant differences were observed in the body lengths of males and females from the two areas. Chromosome substitution analysis Data for egg-adult viability among the substituted strains and the controls are given in Table 2 and illustrated in Figure 1. Analysis of vari¬ ance showed that other than the control strains RX23 and SX23, strains RX3-S2 , R3-SX2 , R2-SX3 and R23-SX had significantly greater egg-adult viabilities at 0.5 ppm AFB^ than the remaining two strains tested. At 1.0 ppm AFB^ , strains R3-SX2, R2-SX3 and RX3-S2 had the greatest egg-adult viabilities. The most sensitive strains at 0.5 and 1.0 ppm AFB^ were RX-S23 and RX2-S3. R23-SX also was among those with the lowest viabilities at 1.0 ppm AFB^. The strains ranked in the same order for egg-pupal viabilities as egg-adult viabilities; however, the differences be¬ tween the strains was not as pronounced. There was no significant differ¬ ence in egg-pupal and egg-adult development times at 0.5 and 1.0 ppm AFB^ in regards to resistance/ sensitivity and chromosomal variation. The body lengths of adult males and females generally decreased with increas¬ ing concentrations of AFB-^. DISCUSSION Data presented in this study show that natural populations of Drosophila melanogaster vary significantly in response to dietary aflatoxin B-j^ and that resistant populations are found in aflatoxin contaminated ar¬ eas. Flies from contaminated areas had not only a greater number of surviving adults when exposed to dietary aflatoxin but also had develop¬ ment times that were significantly shorter than flies from uncontaminated areas. This suggests that natural selection may be a factor contributing the levels of resistance of D. melanogaster to the toxic effects of AFB^. Substituting chromosomes of the sensitive strain (SX23) and the resistant strain (RX23) shows that autosomal factors provide increased resistance to AFBj with chromosome 3 conferring the highest degree of resistance (R3-SX2) . The RX chromosome appears to have a deleterious effect on re¬ sistance to AFB^ toxicity since strains RX3-S2 and RX2-S3 had less resis¬ tance to AFBj than R3-SX2 and R2-SX3. Harmful effects of the RX chro¬ mosome may also account for the observation that strain SX23 had greater egg-adult viability at 0.5 ppm AFB-j, than RX3-S2, RX2-S3, and RX-S23. Strain RX23 did not indicate the presence of a deleterious X chromo¬ some possibly because the RX23 combination may epistatically mask any ill effect produced by the X; or perhaps a resistant X chromosome can en¬ hance resistance to AFB-^ when in combination with R23 and reduce resis- AFLATOXIN SENSITIVITY IN DROSOPHILA 55 Table 2. Effect of larval growth in aflatoxin supplemented media on the number of adults produced for the various chromosomally sub¬ stituted strains. Strain M umber of adults per vial (x ± SE) 0.0 ppm 0.5 ppm 1.0 ppm RX-S23 14.10 + 1.85 1.89 + 1.27 1.00 + 0.0 RX2-S3 13.70 + 2.54 2.66 + 1.22 1.25 + 0.50 RX3-S2 14.20 + 3.55 6.40 + 1.65 4.11 + 1.27 R23-SX 18.44 + 3.00 10.45 + 2.65 2.22 + 1.72 R3-SX2 14.00 + 0.94 12.30 + 3.16 7.90 + 2.85 R2-SX3 16.89 + 2.71 10.20 + 2.14 5.30 + 1.95 RX23 17.30 + 3.20 15.20 + 2.90 12.50 + 4.22 SX23 17.10 + 2.42 9.90 + 2.13 6.50 + 3.17 a. Each treatment was replicated 6 times; each vial initially contained 25 eggs. b. ppm AFB^ in the culture medium. tance when present with either R2-S3 or R3-S2, since R23-SX is less re¬ sistant than RX23 at 1.0 ppm AFB-^. The presence of genes conferring resistance to AFB-^ may cause an overall increase in viability. Under control conditions (0.0 ppm AFBj), strains R2-SX3 and R23-SX had significantly greater egg-adult viability than did the other strains in which chromosomes were substituted; also, under the same control conditions, flies from aflatoxin-contaminated areas had significantly greater egg-adult viability than flies from uncontaminated areas. If selective pressures in the environment lead to an increase in resis¬ tance to AFB-p the question of how Drosophila melanogaster is exposed to the fungus and its toxic metabolites arises. Drought conditions in Virginia during the summer of 1980 contributed to unusually high levels of A. . flavis/ aflatoxin in corn and peanut crops. Many corn samples tested had greater than 400 ppb AFB^ [Personal communication from T. Eadie, Virginia Department of Agriculture] , and A_. flavus was detected in sam¬ ples of peanuts from all the major peanut growing counties [Personal com¬ munication from S. Webb, Virginia Department of Agriculture and consumer Services, Suffolk, VA]. prolonged contact with these crops is not a prima¬ ry mode of exposure of D. melanogaster to aflatoxin. However, a variety of substrates, including fruits in the diet of the flies, may support growth of the fungus. Decaying fruit and other rotting materials, especially when located near contaminated corn and peanut fields, may become infested with 56 THE VIRGINIA JOURNAL OF SCIENCE A. . flavus by airborne spores or insect activity providing a likely route of exposure of D . melanogaster to the toxin. Some fruit flies were trapped in corn, peanut and soybean fields indicating that they can directly become infested with flavus propagules. Pettit et al. [1973] studies the sur¬ vival of A. flavus propagules in peanut soils and showed that the inci¬ dence of _A. flavus remains high in soils from fields on which peanuts are grown continuously and that corn or peanut residues left undisturbed on the soil results in a buildup of A. . flavus populations. D. melanogaster may be exposed to aflatoxin via airborne dust con¬ tamination. Downwind dust samples collected when contaminated corn was being transferred by augers from a storage bin into a wagon and back into the storage bin were analyzed by Burg et al. [1980] and found to have an aflatoxin content range of 12.5 to 204.3 ppb, with an average of 138 ppb. Such dust could settle on surfaces where D. melanogaster rests. Flies tested in this study often were trapped in orchards near a peanut or corn field where harvesting was underway or had been completed. Although this study offers no direct evidence that any of the afore¬ mentioned potential sources of aflatoxin contamination of D. melanogaster actually tranmist the fungus, the fact that resistance of fruit flies to tliic toxin is higher in populations from contaminated areas indirectly indicates that the fungus and its metabolites may become fairly widespread in the environment. Testing the contamination levels of a variety of substrates which may be exposed to natural infestation where and when A,, flavus contamination is high seems highly advisable. ACKNOWLEDGEMENTS We wish to thank Susan Gidley, Pamela PJelone, David Bettinger and Michael Chinnici for helping collect and test the natural population sam¬ ples. This study was supported by a VCU-Grant-in-Aid-to-Faculty awared to J. P. C. LITERATURE CITED Anderson, H. W. , E. W. Nehring, and W. R. Wichser, 1975. Aflatoxin contamination of com in the field. J. Agric. Food Chem. 23(4): 775-782. Burg, W. R. , 0. L. Shotwell, and B. E. Saltzman. 1980. Measurements of airborne aflatoxins during the handling of contaminated com. American Industrial Hygiene Association 42: 1-11. Chinnici, J. P. 1971. Modification of recombination frequency in Drosophila. II. The polygenic control of crossing over. Genetics 69: 85-96. Chinnici, J. P. 1980. Genetic resistance to aflatoxin B^ toxicity in Drosophila melanogaster: chromosome substitution analysis. J. Heredity 71: 275-278. Chinnici, J. P., M. A. Booker, and G. C. Llewellyn. 1976. Effect of Aflatoxin B^ on viabil¬ ity, growth, fertility and crossing over in Drosophila melanogaster (Diptera). J. In¬ verter. Patho . 27: 255-258. Ciegler, A. and J. W. Bennett. 1980. Mycotoxins and myocotoxicoses. Bio Science 30(8): 512-515. Detroy, R. W. , E. B. Lillehoj, and A. Ciegler. 1971. Aflatoxin and related compounds. In A. Ciegler, S. Kadis, and S. Ajl, eds. Microbial Toxins. Vol. VI: Fungal Toxins, Academic Press, New York, 4-154. Dickens, J. W. 1977. Aflatoxin occurence and control during growth, harvest, and storage of peanuts. In J. V. Rodricks, C. W. Hesseltine, and M. A. Mehlman, eds. Mycotoxins in Hu¬ man and Animal Health. Pathotox Publisher, Park Forest, IL. , 99-105. Diener, U. L. , C. R. Jackson, W. E. Cooper, R. J. Stipes, and N. D. Davis. 1965. Invasion of peanut pods in the soil by Aspergillus flavus. Plant Disease Reporter 49(11): 931-935. AFLATOXIN SENSITIVITY IN DROSOPHILA 57 Hesseltine, C. W. , 0. L. Shotwell, W. F. Kwolek, E. B. Lillhoj , W. K. Jackson, and R. J. Bothast. 1976. Aflatoxin occurrence in 1973 corn at harvest. II. Mycological studies. Mycologla 68: 341-345. Hesseltine, C. W. , R. F. Rogers, and 0. L. Shotwell. 1981. Aflatoxin and mold flora in North Carolina in 1977 com crop. Mycologla 73: 216-228. Lalor, J. H., J. P. Chinnici, and G. C. Llewellyn. 1976. Effects of a fungal metabolite, aflatoxin B^ on larval viability and gross morphology in Drosophila melagaster. Develop. Ind. Microbiol. 17: 443-449. Lillehoj, E. B. and C. W. Hesseltine. 1977. Aflatoxin control during plant growth and harvest of com. In J. V. Rodricks, C. W. Hesseltine, and M. A. Mehlman , eds. Mycotoxins in Hu¬ man and Animal Health. Pathotox Publishers, Park Forest, IL, 107-119. Llewellyn, G. C. and J. P. Chinnici. 1978. Variation in sensitivity to aflatoxin B^ among several strains of Drosophila melanogaster (Diptera). J. Inver tebr. Pathol. 31: 37-40. Llewellyn, G. C. and J. S. Katzen. 1981. Aflotoxin occurrence In 1977 com in Virginia. Virginia Journal of Science 32(1): 4-10. Newbeme, P. M. and W. H. Butler, 1969. Acute and chronic effects of aflatoxin on the liver of domestic and laboratory animals: A review. Cancer Res. 29: 236-250. Ong, T. M. 1975. Aflatoxin mutagenesis. Mu tat. Res. 32: 35-54. Patterson, D. S. P. 1973. Metabolism as a factor in determining the toxic action of the aflatoxins in different animal species. Food Cosmet . Toxicol. 11: 287-294. Pettit, R. E., R. A. Taber, and H. W. Schroeder. 1973. Prevalence of Aspergillus flavus in peanut soils. Abstr. American Peanut Research and Education Association Journal 5: 195. Raper, K. B. and D. I. Fennell. 1973. The genus Aspergillus . Robert E. Krieger Publising Co., Inc., Huntington, NY, 686 p. Seiger, M. B. 1966. The effects of chromosome substitution on male body weight in Drosophila melanogaster. Genetics 53: 237-248. Stoloff, L. 1977. Aflatoxins - an overview. In J. V. Rodricks, C. W. Hesseltine, and M. A. Mehlman, eds. Mycotoxins in Human and Animal Health. Pathotox Publishers, Park Forest, IL, 7-28. Wogan, G. N. 1977. Mode of action of aflatoxins In J. V. Rodricks, C. W. Hesseltine, and M. A. Mehlman, eds. Mycotoxins in Human and Animal Health. Pathotox Publishers, Park For¬ est, IL, 24-36. Wong, J. J. and D. P. H. Hsieh. 1976. Mutagenicity of aflatoxins related to their metabolsim and carcinogenic potential. Proc. Natl. Acad. Sci. USA 73: 2241-2244. Virginia Journal of Science Volume 34, Number 2 Summer 1983 Callixylon Wood From The Greenland Gap Group . =4 (Upper Devonian) of Southwestern Virginia Judith E.jSkog Biology Department George Mason University Fairfax, VA 22030 Abstract: A specimen of permineralized fossil wood belonging to the genus Callixylon is reported from the Greenland Gap Group of Upper Devonian age in southwestern Virginia. Study with the scanning electron microscope provided information resulting in a detailed description of the wood and characteristics useful in generic determination. This occurrence extends the geographic range of the genus and documents the presence of fossil axes preserved in pyrite from the area. The genus Callixylon is a form genus for anatomically preserved wood having progymnospermous characteristics and is important as a possible indicator of the presence of the progymnosperm Archaeopteri s . In the United States the genus has been reported from many Upper Devonian localities (Arnold 1931; Beck, 1 962, 1 979; Beck et al . , 1 982; Gillespie et al . , 1981). The Devonian localities range through New York, Michigan, Kentucky, Ohio, Oklahoma, and West Virginia and by reason of this distribution it is not really surprising to find Cal 1 i xyl on in southwestern Virginia. Some Mississi ppian reports of the genus occur as well (Matten, 1972, from Tennessee). The occurrence in Virginia is now documented and the geographic range of the genus is extended. This area represents the southern tip of the eastern arm of the Old Red Continent which lay close to the paleoequator during late Devonian time (Bambach et al . , 1981). Materials and Methods: Three axes preserved by pyrite were found in a sandstone. The locality was discovered and material first collected by Richard K. Bambach at Virginia Polytechnic Institute and State University, 2 km north of locality 11 of Bambach & Kreisa (1973). This new locality is about 5 km north of Blacksburg, VA along highway 460 on the south slope of Brush Mountain. The Upper Devonian beds in this region have tradi tional 1 y been treated as the Chemung Formation (Englund et al . , 1 979). However, Dennison (1 979) has proposed new nomenclature and according to his classification the beds discussed here lie within the Foreknobs Formation of the Greenland Gap Group. The axes were poorly preserved and they fractured very easily. Prepara¬ tion was difficult because of this character. The axes tended to crumble into many pieces if they were not first embedded in Ward's Bioplastic before cutting thin sections on a diamond saw; subsequent coating of each section with epoxy glue and grinding proved the most successful treatment for 1 i gh t microscopy. Axis number 3 (USNM 326722) was prepared in this manner resulting in twenty transverse sections and three longitudinal sections. A portion of axis number 2 (USNM 326721) was also embedded and sectioned resulting in six longitudinal sections. However, the sections still could not be ground per- CALLIXYLON WOOD IN VIRGINIA 59 fectly flat in all areas without some fracturing of the material. The remain¬ ing portion of axis number 2 was split into ten sections for study by light microscopy and by scanning electron microscopy without carbon or metallic coatings. Axis number 1 (USNM 326720) was prepared for scanning electron microscopy by a method described by Grierson (1974): a technique that avoids problems associated with thin sectioning. Some sections were split; some were gold coated and others were left uncoated. The best results were obtained from those split sections that were oxidized in Schulze's solution for 30 to 60 seconds, washed with water, then air dried. The material was mounted on 12 mm round cover glasses with a thin layer of white glue and dried. The cover slips were mounted on stubs with Electrodag 191 SEM glue, coated with carbon and then gold. This double coating eliminated almost all of the charging problems. The sections were observed under one of three microscopes at the National Museum of Natural History, Smithsonian Institution, Washington, D.C.: a Cambridge S4-10, A Cambridge Mark II A, and a Coates and Welter Field Emission Microscope. Speci¬ mens are now deposited at the Museum of Natural History, Paleobotany Collection: USNM 326720 (axis 1), 326721 (axis 2), and 326722 (axis 3). All photographs figured in this paper are from USNM 326720. Descri ption : The fossil material is all secondary xylem with no obvious growth layers as can be seen in the transverse section (fig. 1). The wood is composed of tracheids, 40-50 ym in diameter, and uniseriate or biseriate rays (fig. 1 & 2). The bi- seriate rays may be uniform in width for their entire length except at the ends (fig. 2) or biseriate at the mid-portion only. All rays range from 6 to 20 cells high and are elliptical in tangential section. The ray cells, which are mostly all tracheids, are rectangular (25-40 ym vertically). They are tightly grouped and only an occasional i ntercel 1 ul ar space was observed (fig. 2 & 3) . A variety of pits may occur on the ray cells from bordered to almost simple (fig. 5 & 6) . Cross-field pitting be¬ tween the tracheids and ray cells may be the most variable (fig. 4 & 5). Pits between the ray cells have the least amount of border present. The tracheids have groups of 2 - 10 bordered pits which are arranged in 1 - 2 (rarely 3) rows on the radial walls (fig. 4 & 5) . The pits are circular to slightly oval, 4 - 8 ym in diameter. The apertures appear to be usually aligned; however, debris and pyrite often obscured observation of the orienta¬ tion. Occasionally there is a small pit on the tangential wall of a tracheid, but then only near the side that curves toward the radial wall (fig. 2). 1 The pits in the walls of the tracheids are always bordered to some extent. The borders may be very distinct (fig. 7), or they may be shared by coalescence between adjoining pits (fig. 8). Along the length of a tracheid the pits are always in groups separated by areas of the wall wholly free of pits. The pit groups in adjoining tracheids are aligned radially in uniform rows which is characteristic for the genus Call ixyl on (fig. 4 & 5) . Even though this material was quite fractured and crumbled easily, as can be seen by the extensive debris in the photographs, there are several character¬ istics worth noting particularly. The pits are variable in size and shape on the tracheids. While mainly oval, some are round with extensive borders . Others have a much narrower border surrounding them. The extensive broader borders sometime coalesce resulting in shared pit cavities within the pit groups. Un¬ fortunately no pit membranes could be positively identified in the material from Virginia. Often these shared borders occur in areas of cross-field pitting (fig. 5) but the shared borders also occur between tracheids. Thus no definite correlation of position could be made. Di scussion : The characteris tics of these fossil wood fragments conform with those of the genus Call ixyl on , with particular reference to the consistent grouping of 60 THE VIRGINIA JOURNAL OF SCIENCE Fig. 1. Transverse section of wood showing tracheids and vascular rays (arrows). The factured nature of the material can also be seen with debris from the coating procedure. Photographed at 100 * Fig. 2. Tangential section of wood. Note uniseriate and biseriate rays, pitting of tracheids with occasional pit on the tangential wall (arrow). Photographed at 120 x Fig. 3. Radial section of wood showing ray and grouped pitting of all cells (arrows). Photographed at 200 CALLIXYLON WOOD IN VIRGINIA 61 Fig. 4. Radial view of the interior of tracheids enlarged to show grouped pitting and narrow borders of pits. Photographed at 360 * Fig. 5. Radial view of tracheids with ray immediately below them to show cross field pitting. Shape of pits varies from circular to oval. Borders within some groups raised (center of photo). Photographed at 600 * 62 THE VIRGINIA JOURNAL OF SCIENCE Fig. 6. Ray tracheid showing pits with narrow borders. Photographed at 1000 x Fig. 7. Circular bordered pit of tracheid with wide border. Photographed at 3000 * Fig. 8. Bordered pits of tracheid with coalesced borders. Photographed at 2600 * CALLIXYLON WOOD IN VIRGINIA bordered pits in the secondary walls of the tracheids. The common occurrence of the genus as isolated axes in Upper Devonian sediments also lends some additional credence to the positive identification of the fragments described herein. All three axes seem to represent the same taxon. While the genus Cal 1 ixyl on is better identified when primary growth is also preserved the generic name has been unhesitatingly applied to fragments of secondary wood (Beck, 1962; Matten, 1972). Other form genera, such as Desmoporoxyl on are in need of better descriptive detail before comparison can be made with the specimens described here. The species included in Cal 1 ixyl on are difficult to distinguish because of considerable variability and resultant overlapping of characters. Arnold (1931) in cautioning against establishing new species of Cal 1 ixyl on noted that variations in tracheid diameter, the number of pits per grouping, the contents of ray cells, and presence or absence of growth rings are not re¬ liable species characters. Beck et al . (1982) also describe the difficulty of identifying Call ixyl on to species. Thus the specimens described here are simply reported as Cal 1 ixyl on sp. As more of the Cal 1 ixyl on material from the locality cited here is studied with the scanning electron microscope better specific assignments may be made. The technique of preparation for the scanning electron microscope proved to be advantageous. Coating of the specimens with carbon followed by gold practically eliminated problems of charging even though more debris developed from and around the specimens as a result of the longer period under vacuum. As a result of the disintegration of the specimens more cell detail could be examined than when no coating at all was applied or only a coating of gold was used. This material described and illustrated here is significant because of the extension of the geographic range of Call ixyl on, the documentation of the occurrence of pyritized plant material in the Upper Devonian of southwestern Virginia, and the additional study of specimens of Call ixyl on with the scan¬ ning electron microscope. Acknowl edgements : The author thanks Dr. Francis M. Hueber for the use of his laboratory facilities at the National Museum of Natural History and Mr. Jan Endlich for aid in preparing the figures. Literature Cited Arnold, C. A. 1931. On Call ixyl on newberryi (Dawson) Elkins et Wieland. Contr. Mus. Paleont. Univ. Michigan III: 207-239. Bambach, R. K., C. R. Scotese and A. M. Zeigler. 1980. Before Pangea: the geographies of the Paleozoic world. Amer. Sci . 68: 26-38. Beck, C. B. 1 962. Plants of the New Albany Shale. III. Cal 1 ixyl on . Rev. Palaeobot. Palynol . 28: 103-115. Beck, C. B., K. Coy and R. Schmid. 1982. Observations on the structure of Cal 1 ixyl on wood . Amer. J. Bot . 69: 54-76. Dennison, J. M. 1970. Stratigraphic division of Upper Devonian Greenland Gap Group ("Chemung Formation") along Allegheny front of West Virginia, Maryland, and Highland County, Virginia. Southeastern Geol . 1 2: 53-82. Englund, K. J., H. H. Arndt, and T. W. Henry (eds.). 1979. Proposed Pennsylvanian System Stratotype Virginia and West Virginia. Amer. Geol. Inst. Selected Guidebook Series no. 1. 138pp. 64 THE VIRGINIA JOURNAL OF SCIENCE Gillespie, W., G. Rothwell and S. Scheckler. 1981. The earliest seeds. Nature 293: 462-464. Grierson, J. D. 1976. Leclercqia complexa (Lycopsida, Middle Devonian): its anatomy, and the interpretation of pyrite petrifactions. Amer. J. Bot. 63: 1184-1202. Kreisa, R. D. & R. K. Bambach. 1973. Environments of deposition of the Price Formation (Lower Mississippian) in its type area, southwestern Virginia. Amer. J. Sci., Cooper Vol . 273-A: 326-342. Matten, L. C. 1 972. Cal 1 ixyl on from the Maury Formation (Lower Mississippian of Tennessee. J. Paleontol. 46: 711-713. Virginia Journal of Science Volume 34, Number 2 Summer 1983 Prevalence and Distribution of Lungworms of Swine Raised in Small Farm Units of Southside Virginia Dilip K. < Sen, Euzebio B.jDamazio and Peter J. McN^rney Parasitology Laboratory Department of Life Sciences Virginia State University Petersburg, VA 23803 ABSTRACT The prevalence of helminths was determined frcm swine fecal materials collected frcm 136 small farm units of Southside Virginia. The survey was conducted during the suirmers of 1980 and 1981. Out of the twenty-seven counties surveyed, fecal samples from seven revealed the presence of lungworm (M tno ng ytuA 4pp. ) infection in pigs. The high¬ est percentage prevalence of lungworm of swine was recorded as 35.0% of the twenty samples analyzed for Charles City, followed by 16.6% of the twelve samples examined frcm Mecklenburg County and 10.8% of the thirty- seven fecal samples analyzed from Powhatan County. The overall lungworm egg counts as revealed by fecal analysis frcm all the twenty-seven count¬ ies were low (3%) and sporadic. The observation of such low incidence of swine lung-worm infection is not uncommon as other researchers frcm the Southern states, especially frcm the mid-atlantic region, reported similar findings. The prevalence of helminths appeared to be related to the overall quality of management. INTRODUCTION | I Lungworms [Mztcut/iongy-tLU 4pp.) are most frequently present among hogs raised in open hog lots (Alicata, 1964) . Most metastrongyles are called 'lungworms' and they usually inhabit the respiratory or circula¬ tory systems of carnivores, ungulates, rodents and primates throughout the world. Numerous reports indicate the presence of lungworm infection in the United States. When the species have been identified, both MzAtaAt/iongylui zZongatuA and M. Pude.ndotectiL6 have usually been found together in mixed infestations, the former species usually being seen more frequently and in greater numbers than the latter, although Ledet and Greve (1966) found both species present in almost equal numbers in swine. Ewing and Todd (1961) also found M. 4 cUmi. present in mixed infestations, but only one percent of the infected pigs carried this species (Rose, 1973) . Light to moderate infections cause no marked clinical symptoms in pigs, but heavy infections can cause pneumonia and other respiratory disease and may cause death as mentioned by Sullivan and Shaw (1953) . 66 THE VIRGINIA JOURNAL OF SCIENCE Several workers have described the pathology of both natural and experi¬ mental swine lungworm infections in varying degrees of detail (Hollow, 1966; Tiunov, 1966; and Rose, 1973) . The value of swine marketed in North Carolina alone has increased from 96 million dollars in 1965 to 247 million dollars in 1974 and it was projected that by the eighties, 10 million heads of swine will be produc¬ ed in the coastal plains regions of Virginia, North Carolina and South Carolina. According to the USDA (1965) , swine lungworms cause an annual loss of $3,584,000 in the U.S.A. , of which $171,000 is due to death and the remainder to illness. These facts coupled with (1) lack of data of these worms in definitive hosts from small farm units in Southside Virginia and (2) the changes in this region's population, trade and socio-economy indicated a need for up to date information on this impor¬ tant parasite of pigs. MATERIALS AND METHODS Trips were made to small farm units in twenty-seven counties and three feeder pig sales in Southside Virginia. A total of 136 farms were visited during the summer months of 1980 and 1981. Freshly discharged pig feces from the identified animals were used for analysis. Disposable boots and gloves were used to prevent the transfer of parasite ova from one farm to another. Fecal samples were placed in polyethylene specimen bags and refrigerated at 7°C for not more than 3 days prior to analysis. The method found to be reliable and most satisfactory for analysis of helminth ova is Sheater's Sugar Flotation Technique. The solution has a specific gravity of 1.2 and is most useful when looking for nematode eggs (Palmier i and Anderson, 1979) . A concentrated solution was made by dissolving 390 g of sugar in 250 ml of hot (90OC) water. Once cooled, 4.6 g of phenol was added to the solution to deter mold growth. The concentrated stock solution was thoroughly mixed 1:1 with tap water at room temperature just before use. Two g of fecal sample was placed in large crucible with 20 ml of the 1:1 sugar /flotation solution and thoroughly mixed until suspended. The mixture was poured through cheesecloth and the remaining debris was discarded. The suspension was centrifuged at 200g for 5 minutes in a clinical centrifuge. The centrifuged material was allowed to stand for 5 minutes before analysis. A special glass rod with a flat head was dipped into a centri¬ fuge tube until it just touched the upper layer of the mixture. The head of the glass rod containing the specimen was then touched to a drop of water placed in a 3x1 inch glass slide and covered carefully with a coverslip to avoid trapping of air bubbles. Finally, the parasite ova were identified and measured under the microscope equipped with a micro¬ meter. The identification of lungworm ova was based on the description outlined by Morgan and Hawkins (1960) . RESULTS AND DISCUSSION Three percent of the total 568 swine fecal samples collected from pigs of all ages from small farm units of Southside Virginia were positive for lungworm infection. This survey showed that the incidence of lung¬ worm infection was 35% of the twenty samples examined from Charles City County, followed by 17% of the eleven samples examined from Mecklenburg County and 11% of the thirty-seven samples analyzed from Pcwhatan County. Other counties with 3 to 10 percent prevalence of lungworm were Appomatt¬ ox, Dinwiddie, Sussex and Greenville counties (Table 1). Sixty five swine fecal samples also collected from three other feeder pig sales at Farmville, Southampton and Tappahanock of Southside Virginia were found to be negative for lungworm infection. Fecal egg counts were determined. The lungworm egg counts were low and sporadic. This was not considered unusual, as there is a scarcity of LUNGWORMS IN VIRGINIA SWINE 67 scientific reports linking metastrongyle egg counts to clinical disease (Dunne, 1959) . The incidence of lungworm infection as recorded by different work¬ ers varies considerably. Andrews and Connelly (1945) in Georgia, Batte and Moncol (1966) in North Carolina, and Riddle and Forrester (1972) in South Carolina found a low incidence of lungworm infection (Table 2) . Kelly and Sen (1959) in Nebraska and Nevenic and Sibalic (1953) in Yugoslavia also found a low incidence of swine lungworm infection. Swine lungworms are pathogenic both in their own right and because they may transmit or exacerbate certain diseases. Loss of condition and retarded growth are common results of infection, and young pigs may be killed by heavy infections (Levine, 1980) . The United States Department | of Agriculture has estimated the annual loss due to harmful effects caused by internal parasites of swine to be more than 275 million dollars (Stewart and Southwell, 1960). A large portion of this loss occurs in the southern states. In North Carolina, for example, internal parasites reduced the value of swine by at least one dollar for each pig marketed (Batte and Moncol, 1966) . The control of swine parasites, therefore, is a major problem, particularly since intensification and expansion of the swine industry has not been accompanied by the reduction of worm popul¬ ations (Batte et al., 1965). The pigs raised on small farm units of Southside Virginia, in this survey, were subjected to the most primitive forms of swine management. Animals were kept in dirt pens that were at best irregularly cleaned and remained shaded and moist throughout the year. However, a few farms had access to open pasture. Most of the farmers were primarily of the lower socioeconomic category and were part-time swine producers. They placed little or no emphasis on housing, nutrition or health of their pigs and were unaware of modem swine management practices. Swine lungworms ( MeXcui&iong ytii6 4pp.) were detected in Charles City, Mecklenburg, Powhatan, Appomattox, Dinwiddie, Sussex and Greenville counties of the Southside Virginia. The majority of farmers in these counties kept their animals in open hoglots where the intermediate (earth¬ worm) hosts were present in great numbers. Since the earthworms are the Table 1. Percentage prevalence of MeXaAt/iongyZuA 4pp. of swine in Southside Virginia as determined by fecal analysis during 1980-81. County Locality No. Infected/ No. Examined Percentage Infected Charles City County Southside Virginia 7/20 35.0 Mecklenburg County II 2/12 16.6 Powhatan County II 4/37 10.8 Appomattox County II 2/20 10.0 Dinwiddie County II 2/24 8.3 Sussex County II 1/22 4.5 Greenville County II 1/33 3.0 Remaining 20 Counties 91 0/400 0.0 Total 19/568 3.3 68 THE VIRGINIA JOURNAL OF SCIENCE Table 2. Comparison of percentage prevalence of Mutcu&iongyZuLA 6 pp. of swine in Georgia, North Carolina and South Carolina with the present study (Southside Virginia) Investigators Percentage Pre Valence Area (years of analysis) Andrews and Connelly 1945 14 Georgia (1943-44) Batte and Moncol 1966 0 North Carolina (1965-66) Riddle and Forrester 1972 5 South Carolina (1968-69) Sen et al. , 1983 3 Southside Virginia (1980-81) intermediate hosts of swine lungworms , measures which prevent pigs frcm rooting in the ground and eating earthworms will prevent lungworm infect¬ ions. If possible, hogs could be kept on concrete and their feces should not be spread on pastures where pigs will later run (This ideal situation prevailed in 20 counties where lungworm infections were not prevalent) . Good sanitation and adequate nutrition will go a long way to protect pigs, raised in Southside Virginia, frcm parasites. No drug will remove all the internal parasites (although specific chemotherapeutic drugs are available for swine lungworm infection) , and certainly, the damage done by the migrating worms cannot be cured (Stewart and Hale, 1978) . A good system of management will incorporate practices aimed at the prevention of infection and will not use treatment as the sole method of controlling parasites . ACKNOWLEDGEMENTS The authors wish to express their deep appreciation to the County Extension Agents of Southside Virginia for their cooperation in making this investigation a success. Thanks are extended to Mr. J. Okoye and Mr. A. Tsige of Virginia State University for their active help during the investigatory process. This study was supported by a grant #801-15-10D frcm the United States Department of Agriculture (CSRS/USDA) and a grant #S06-RR-08090-10 frcm the National Institutes of Health (MBRS/DRR) which are acknowledged with pleasure. Special thanks are due to Dr. H. Hamlett and Dr. V. Purohit for revision of the manuscript. LITERATURE CITED Alicata, J. E. 1964. Parasitic infection of man and animals. Hawaii Agri. Exp. Sta. Univ. of Hawaii. Tech. Bull. No. 61. Andrews, J. S. and J. W. Connelly. 1945. Worn in swine raised under a moderate degree of sanitation. Proc. Helm. Soc. Wash. 12: 6-12 LUNGWORMS IN VIRGINIA SWINE 69 Batte, E. G. and D. J. Moncol. 1966. Internal parasites of swine. North Carolina State Univ. Anim. Sci. Rep. 171. Vet. Ser. 7: 1-15 Batte, E. G. , D. J. Moncol., A. C. Todd and R. S. Isenten. 1965. Critical evaluation of an anthelmintic for swine. Vet. Med. Small Ani. Clin. 60: 539-545. Dunne, H. W. 1959. Diseases of swine. Iowa State Univ. Press. Ames. Ewing, S. A. and A. C. Todd. 1961. Metastrongylosis in the field, species and sex ratio of the parasites, preferential location in respiratory apparatus of the host, and concomitant lesion. Am. J. Vet. Res. 22: 606-609. Hollow, R. 1966. Investigation on metastrongylosis in swine. III. The pathogenicity of lungworms in artificially infected animals. Acta. Vet. Hung. 16: 413-428. Kelley, G. W. and H. G. Sen, 1959. The incidence of swine lungworm in Nebraska. J. Parasit. 45 (4 sect. 2) : 41. Ledet, A. E. and J. K. Greve. 1966. Lungworm infection in Iowa swine, J. Am. Vet. Assoc. 148: 547-549. Levine, N. D. 1980. Nematode Parasites of Domestic Animals and of man. Burgess Publishing Company. Minneapolis, Minn. 246-247 pp. Morgan, B. B. and P. A. Hawkins . 1960. Veterinary Helminthology. Burgess Publishing Company, Minneapolis, Minn. 177-180 pp. Nevenic, V. and S. Sibalic. 1953. Contribution to knowledge of the spread of pulmonary parasites of swine. Vet. Glasnik . 7: 176-181. Plamieri, J. R. and F. L. Andersen. 1979. Parasites in dog feces. Carolina Tips. No. 42. Burlington, North Carolina. Riddle, D. and D. J. Forrester, 1972. Prevalence and distribution of helminths of swine in South Carolina. Proc. Helm, Soc. Wash. 39: 55-59. Rose, J. H. 1973. Lungworms of the domesticated pig and sheep. Adv. Parasitol. 11: 559-599. Stewart, T. B. and O. M. Hale. 1978. Management and treatment of wormy pigs. ARS/USDA and Dept. Anim. Sci. Resch. Bull No. 214. 1965. Stewart, T. B. and B. L. Southwell. 1960. The economic importance of internal parasites of swine and their control by management and treatment. Ga. Coastal Plain Sta. Mimeo. 102: 1-13. Sullivan, J. R. and J. N. Shaw. 1953. Incidence and effect of lung¬ worm in Oregon swine. Oregon Agri. Exp. Sta. Tech. Bull. 23:11. Tiunov, I. 1966. Inmunity in porcine metastrongylosis. Vses. Inst. Gel'mint. 12: 205-211. U. S. Dept, of Agriculture. 1965. Losses in Agriculture. Ag. Handbook #291. USDA, Washington, D. C. 120 pp. Virginia Journal of Science Volume 34, Number 2 Summer 1983 (A Computer Data Acquisition and Analysis Program for the Laboratory Russell W. Vaughan and David A. Jenkins Department of Physics Virginia Polytechnic Institute and State University Blacksburg, VA 24061 ABSTRACT An available software system for laboratory data collection is described having broad application in computer assisted experimentation. Program features are described for data analysis, and hardware considerations are outlined with recommendations for program utilization with or without program code modification. The intent is to acquaint potential experimenters with the fundamental considerations of such a project by example. 1. The Expansion of Computer Utility Electronic computational devices continue to have increasing influence on daily life. Commonly these devices appear in specialized products such as calculators, toys, clocks, stereo equipment and televisions. In these cases the computational function is fixed or the user has little control over modifying the dedicated nature of the computer. In a more limited extent, the truly all purpose, totally programmable computer is also finding wider acceptance until ultimately domestic computers may be commonplace. The computer has become an indi spensible tool in many aspects of data collection and analysis. The increased accuracy and productivity afforded is no longer a matter of convenience, but of necessity in conducting ever more complex experimental procedures. The alleviation of complexity in performing the experiment by introduction of a computer actually only shifts the complexity into the design stages. But, once the design is finalized, an experiment may be conducted with a high degree of precision and repeatability . In the design one must consider the interfacing of computer to experiment and establishment of a means for manual and automataic control. Decisions must be made as to which functions will be provided by hardware or by software MULTI FOR LAD DATA ANALYSIS 71 Fig. 1. Components of typical computer assisted experimentation. SHOU DISTRIBUTION - RUN 0 Fig. 2. The graph called ‘show’ exemplifies the MULTI histogram support feature for data analysis. 72 THE VIRGINIA JOURNAL OF SCIENCE (programs) and extensive testing of both together and separately must be done. One will inevitably find that while hardward is diverse in selection and reliable in operation, implementation of software is primarily a user's responsibility. Software design often becomes the most expensive phase of the project and, if not implemented properly, can easily hinder progress. This paper has been prepared to acquaint a potential experimenter with the overall problems and advantages of using computers and to describe a program called MULTI for use directly or as a model in software pre paration. 2. Applicability The Fermilab National Accelerator research facility at Batavia, Illinois has created a computer program named MULTI functioning as a data acquisition and analysis tool for laboratory experimentation. MULTI ' s intentioned application is within particle physics, however the program is of sufficient generality to recommend itself to other unrelated applications . We present here a description of the general role a computer may fill in experimental situations, the functions MULTI provides and a suggestion for using MULTI in the general laboratory situation. a. Experiment Hardware The fundamental aspects of a computer assisted experiment are summarized in figure 1. Three main components comprise the minimum design. One component, the experiment itself, produces data to be communicated into the computer, the second component. In more sophisticated systems the flow of data can be reversed from computer to experiment. This provides for automatic control through feedback paths. A third component, the computer to experiment interface, is responsible for this uni- or bi-directional data path. It provides for translation of data and control signals into analog or digital forms appropriate to the hardware. The computer alone is made up of several modular parts to function as the human to machine interface. A terminal upon which an operator enters commands and receives responses is necessary for interactive control. This same terminal or a separate device should be capable of displaying graphical data. Some form of storage such as a magnetic tape drive is a necessity for recording voluminous data. Typically, a spinning magnetic disk known as a direct access storage device is used to store various programs and small data files . b. Experiment Software An integral part of the system is the program running it. An insufficiently designed software system can destroy the performance of the best hardware. Programs can be cryptic. MULTI FOR LAB DATA ANALYSIS 73 inconvenient and unreliable. Too often programs are thrown together for immediate results and the cost of such practice later becomes evident. Software design is best when it assumes simplicity, has a logical organization, and assumes an analytic role during execution. The analytic function is exemplified by such practices as coding into programs a dialogue to explain commands, verify data, and confirm user intentions at critical experiment stages. This feature can be helpful in preventing potentially destructive command sequences from being inadvertently entered. The complementary function that software fills in experimentation might include the following attributes: - can incoming data be easily manipulated through keyboard commands? - can data be automatically monitored and appropriate action taken? - can data be displayed graphically as they occur? - are the data recorded and catalogued properly with easy access? - is the system expandable on the user and programmer levels? - is the documentation describing the system adequate for novice users? The above mentioned qualities are general enough to apply in most computer assisted experiments. Indeed they are the major aspects of Fermilab's MULTI software which is specifically discussed below. 3. MULTI Operation Human to machine communication in interactive systems is commonly accomplished through commands entered on a keyboard and responses displayed on a television screen device. These commands usually take the form of a keyword and parameter list where the keyword invokes a basic function and parameters determine specific actions within it. A sample of the communication scheme taken from MULTI will illustrate both the programming technique and a facility of MULTI. a. A Sample MULTI Session MULTI is useful over the full range of an experiment's duration from design, to control, to indepth analysis. If experiment hardware is not yet available, the software development may proceed with a simulation method of data collection provided by MULTI. When data have been recorded it is possible to replay the collected data and analyse them extensively. The following command/response sequence illustrates several MULTI capabilities. It is responsible 74 THE VIRGINIA JOURNAL OF SCIENCE for producing the graph photographically from a terminal. The > sign is MULTI will accept operator of figure 2 which was taken Tektronix 4010 vector drawing a prompting character indicating commands . >SET DATA=MT The user indicates that an experiment playback from magnetic tape is to occur. >HI ST DA6 , SHOW, 1024,0,1023 The keyword HIST indicates that a histogram is to be constructed from entering data. The following parameters respectively establish where data are to be found in memory (location DA6), a histogram name for operator reference (SHOW), bin count (1024), originating and terminating bin values ( 0 , 1023 ) . >D I SPLAY SHOW The previously defined histogram is displayed on a graphics device . >C0NTINUE This initiates reading of a tape. As data enter MULTI, the histogram is updated internally and because it is under active display these updates are immediately conveyed to the graphics device. A number of histograms may be defined simultaneously and called up for individual display during data acquisition. b. The Practical MULTI Consideration must now be given to the practical aspects of implementing MULTI on real hardware. MULTI ' s requirements vary depending on the desired function of MULTI itself, however a minimum system would include a Digital Equipment Corporation' s LSI-11 computer (Digital Equipment Corp oration, Maynard, Mass., 01754 with 28K words of memory and enough external disk storage (about two megabytes) to generate MULTI. The user generates MULTI by selecting various subroutines then compiles and links them into a runnable program. A graphics device, printer and magnetic tape unit are highly desirable but not necessary to run limited versions of MULTI . MULTI FOR LAB DATA ANALYSIS 75 Fig. 3. The CAMAC crate functions as an interface of experiment to computer. It is a device on the common external device control channel as are other hardware components such as disk and tape drives. 1.0 MULTI 0.5 MULTI Fig. 4. The relative functional differences between MULTI versions 1.0 and 0.5 are only those of data acquisition capability. User versions of MULTI are possible by modifying the 0.5 system. 76 THE VIRGINIA JOURNAL OF SCIENCE c. Different MULTIs The user puts together a MULTI configuration according to the hardware available. The most capable MULTI is also the most specific in hardware requirements. The user may feel compelled to attempt code modification for personalized versions of MULTI utilizing hardware unique to the installation. This would be done by selecting the standardized version of MULTI most appropriate to the application and then building upon it. There exist two documented versions of MULTI called the full and the half systems designated 1.0 and 0.5 respectively. A full system provides the capabilities as described above for general laboratory setups using very specific hardware. Since the partial system, as a subset of the full one, provides only the analysis function, hardware requirements are less stringent. We first consider intricacies of MULTI 1.0 in the laboratory environment. d. The CAMAC Crate To fulfill the function of the computer to experiment interface, the level 1.0 MULTI is written utilizing a CAMAC crate as seen in figure 3. CAMAC (for Computer Automated Measurement and Control) is a standardized interface of computer to a wide range of modular plug-in devices useful in experiments to perform such functions as analog to digital conversion, digital to analog conversion, pulsing, timing, counting, stepper motor control and interrupts. It was developed and is widely used by high energy physicists. The software necessary to utilize the device is available within MULTI 1.0 in two forms. First, there is a convenient sub- language supported by MULTI in which the user programs CAMAC module servicing code. When an event of interest occurs, the computer is directed to the CAMAC code which typically would read out data and reset the experiment in preparation for another event. Secondly, a well documented set of subroutines exist for controlling CAMAC for the FORTRAN level programmer independent of MULTI. It must be indicated that MULTI 1.0 requires a special crate control module known as the JY411(Jorway Corp., 27 Bond St., Westbury, N.Y., 11540). Obviously the full system is quite hardware dependent in the computer to experiment interface aspect. Unless you have access to the proper CAMAC hardware, MULTI 1.0 is of no use. Additionally, MULTI 1.0 expects to find special FERMILAB designed hardware on line. Although a laboratory may possess the specified CAMAC equipment, it is still necessary to disable that part of MULTI making reference to unique Fermilab hardware. Figure 4 shows the relationship of MULTI 0.5 to 1.0 in terms of specific hardware requirements. It is seen that the basic difference is simply that of data gathering ability. This makes MULTI 0.5 a simpler analysis only system. It also suggests that MULTI 0.5 is the best candidate for adaptation to particular laboratory needs. MULTI FOR LAB DATA ANALYSIS 77 e. MULTI 0.5 with Magnetic Tape The simplest implementation of MULTI would be to use it in unmodified form. Supposing you have already developed data acquisition and recording programs, it would be a relatively easy task to take the recorded data and reformat them into a MULTI style tape. This would amount to simulating the acquisition of data as if MULTI had originally collected and recorded them. The resulting data are then available to a 0.5 MULTI for full analytical study on playback. The structure of such tapes is fully documented in Fermilab publications accompanying MULTI. A programmer familiar with writing for tape devices should be able to produce a successful reformatting program. If such off-line analysis is acceptable, this is the route to follow. f. MULTI 0.5 with an Experimental Interface If it is desired to have MULTI ' s analytic capabilities running in parallel with the experiment, a slightly more complex procedure of code modification is necessary. The basic strategy we followed was to exploit the tape reading interface of MULTI in the playback (tape reading) mode. The tape record, as explained in the documentation, is simply a string of binary values grouped into computer words and preceded by a few values of a descriptive nature as to how to use these data. Certainly, a real time sequence of experimental data could be made to look like a magnetic tape record. This would be accomplished by buffering the incoming data into storage and adding onto the front of them the tape record leader information. When a record is complete MULTI can be signaled that data are available. The task to be confronted, then, is grafting into MULTI one's own acquisition code and presenting the accumulated data to MULTI in the form of a simulated tape record. Specifically, the tape read instruction of MULTI code is replaced with code to transfer the collected data to a specific buffer area of memory for MULTI processing. 4. The Structure of MULTI Software The logical structure of MULTI may be likened to walking a circular hallway lined with doors. As one walks the floor he tries each door. If the door opens he enters to investigate and later returns, closing the door behind him. A locked door is ignored and his walk continues. Each door represents a condition of the system such as a command entered, or a tape record available. The door lock is a flag variable tested by the program whose flow is altered according to the test. A door unlocked is a flag variable set and attention is provided accordingly. This design persuades and suggests to a programmer that, in the adaptation of MULTI to a unique setting, he investigate the methods of data transfer through a convenient channel such as in the tape reading part of MULTI . MULTI 1.0 is an interrupt drive program. An interrupt is a non-scheduled, random occurence of some event external to the computer which demands immediate attention from a special segment of the software. When an interrupt occurs 78 THE VIRGINIA JOURNAL OF SCIENCE this special code is called up to perform a task as specified by the programmer. In the hallway analogy an interrupt occurs when a door pops open and the walker rushes to enter, ignoring all others. When through, he resumes the walk at the point of interruption. The acquisition code to be grafted onto MULTI 0.5 can also be interrupt driven. It is in this interrupt code that the simulated tape record is constructed from experimental data and a flag variable is set (a door is unlocked) to initiate an artificial tape read. Alternatively, if the computer to experiment interface is not interrupt driven, a device polling strategy of programming style could be accomodated. In the walker analogy this amounts to adding another door. The programming style adopted depends on what kind of computer to experiment interface is available. The important aspect of the problem is to study the original tape reading MULTI code and devise a method for substituting real time data for tape records. 5. Extensions to MULTI In our own versions we have made numerous extensions to MULTl's capabilities in keeping with our own hardware configuration. Certain areas of memory have been added or extended to make esoteric data accessible through special keyboard commands. We have MULTI s to work with three kinds of CAMAC crate configurations on three different DEC computers. We have added to MULTI a means of producing selected color graphics on a Ramtek 6000A terminal so that up to 8 histograms may be viewed simultaneously or 4 individually as they are updated. This extended MULTI is implemented on a PDP 11/34 with two model RL01 disk drives using a JY411 controller in a CAMAC crate. Currently the system is used at our experimental site of the Indiana University Cyclotron Facility at Bloomington, Indiana. Acknowledgements We thank T. Lagerlund and L. Taff for their assistance in adaptation of MULTI to our specific requirements. This material is based upon work supported by the National Science Foundation under grant PHY-7828080. Literature Cited IEEE Transactions on Nuclear Science, CAMAC Tutorial Issue, April 1971, Vol. NS-18, Number 2., 105 pp. L. Taff, D. Ritchie, T. Lagerlund., 1979., RTMULTI Data Acquisition System, Fermilab Publication PN-110.1K, 46 pp. L. Taff, D. Ritchie, E. Quigg, T. Lagerlund, 1980., Fermilab MULTI User's Guide, Fermilab Publication PN-97.5, 65 pp. E. Quigg, T. Lagerlund, D. Ritchie, L. Taff, 1980., MULTI MULTI FOR LAB DATA ANALYSIS 79 Programming Reference Notes, Fermilab Publication PN-106, 85 pp. L. Taff, D. Ritchie, T. Lagerlund, 1979., RTMULTI System Generation, Fermilab Publication PN-98.1K, 41 pp. Virginia Journal of Science Volume 34, Number 2 Summer 1983 u Distribution of Pinus virginiana and Pinus taeda in the Peninsula of Virginia ^ f Cindy Larson Rice and StewartjWare Department of Biology College of William & Mary Williamsburg, V A 23185 Abstract Virginia (Pinus virginiana) and loblolly (P. taeda) pines are both abundant between the James and York Rivers in Virginia. Studies by R. Monet te and S. Ware found loblolly pine to be overwhelmingly dominant in old field succession, with Virginia pine absent or rare even when seed trees of that species were nearby. In our survey we found the same results, and found Virginia pine to be abundant in areas that had been heavily (selectively) timbered (perhaps by cutting out an earlier generation of pine) , had been clearcut but not further managed, or had been bulldozed off so that the red topsoil was exposed (sloping road cuts, for instance). Loblolly pine ranged from almost as abundant (usually) to absent (rarely) in these same sites. Despite the clear association with past land use, neither pH, Ca, Mg, K, P soil textures or growing season moisture (measured at 2-wk intervals with Bouyoucos moisture meter) correlated with abundance of the two species. Introduction Virginia pine (Pinus virginiana) and loblolly pine (Pinus taeda) are both abundant trees in the forests of the central Coastal Plain of Virginia. However , Monette & Ware (1983) found in a study of old field succession in the area that the early forest stage was overwhelmingly dominated by loblolly pine, with Virginia pine absent or insignificant. In a survey of over 40 pine stands, we likewise found that old field pine stands had very little Virginia or shortleaf (P. echinata) pine, even when older seed trees of these species were nearby. On the other hand, stands in which Virginia pine attained dominance or co-dominance with loblolly pine were either bulldozed road cuts with the red lateritic subsoil exposed, or more often, cut-over areas with varying degrees of previous timbering. Areas which had been selectively cut (for a previous generation of pine) had more relict hardwoods mixed in with the pines than did areas that had been clearcut, but the ratio of the two major species of pine did not seem to be affected by the degree of previous timbering. McQuilkin (1940) has described the importance of seed source in establishment of pine stands, rating this factor higher than effects of normal variations in site, soil type and plant cover. However, if the DISTRIBUTION OF VIRGINIA PINE 81 Table I. Species Composition (density %) of seedling stands and probable seed source stands. Seedling Source Stand Stand Site 1 P. taeda 15.5 11.1 P. virginiana 84.5 88.9 Site 2 P. taeda 19.5 8.3 P. virginiana 69.6 75.0 P . echinata 11.0 16.7 assumption is made that adequate seed supply of both species was available at the time the fields were abandoned or the woods cut, seme other answer must be sought to explain why Virginia pine is important only on cut-over areas, usually in co-dominance with loblolly pine; and is unimportant in loblolly- dominated abandoned fields. This study was undertaken to examine several factors that might be involved in this differential distribution of the two pines. Methods This study was made in the area of Williamsburg, Virginia beginning in May 1976. The number of pine stands available for study was limited by local farming or construction activities and the resultant destruction of pine woods; in all, eight stands were selected for their lack of recent (25-30 yrs) disturbance. Stands 1 and 2 are separated from each other by a small hardwood valley; stands 3 and 4 are located on opposite sides of a two- lane roadway and are sufficiently different from each other to be considered separate stands on the basis of species conposition. None of the other stands were near each other. All stands were approximately equal in size (2-3 ha) and age (25-30 yr) based on the larger pine trees. Sampling was done with a Spiegel Relaskop (Bitterlich method) and circular quadrats or by the point-centered quarter method. Four of the stands obviously originated on abandoned fields, as traces of crop rews were still evident. The remaining four stands had come in on cut-over areas, indicated by numerous rotted stumps. Stands 3, 4, 7, and 8 were judged to be old field while stands 1, 2, 5, and 6 were cut-over in origin. In addition to these 8 stands two 10-yr old pine stands were studied in an attempt to determine whether seed source controlled species composition in non-old field sites. Each of these stands was long (50 m and 70 m) and narrow (- 5m) and located on fairly steep road banks Which had originally been bulldozed down to the red lateritic subsoil. Each community bordered a mixed species cut-over woods (larger pines 5CH- yrs). In each strip community all pine seedlings were tallied according to species. All cone-bearing pines in the adjoining forest were tallied as probable seed sources if their crown extended out over part of the young pine stand or if their trunk was within 5 m of the edge of the young pine stand. In the 8 sampled stands, analysis was made of several edaphic factors which could be responsible for favoring the occurrence of either species on 82 THE VIRGINIA JOURNAL OF SCIENCE a site. Soil moisture as recorded at two-week intervals beginning in May 1976 and continuing through early November 1976, using a Bouyoucos moisture meter and gypsum blocks buried at 15 cm depths at scattered locations within a stand (but always within 0.5 m of a loblolly pine in the loblolly stands and within 0.5 of a Virginia pine in "mixed" s tands ) . Several soil samples were taken at each site. Litter and duff were removed and a cylindrical sample was taken to a depth 15 cm. All samples from a stand were blended and divided into two portions. Soil texture analysis was performed on one portion according to the method described by Bowers (1976) . Results were recorded in percent values of sand, silt, and clay. The other portion was sent to the USDA Cooperative Extension Service Lab at Virginia Polytechnic Institute and State University for analysis for calcium (CaO) , magnesium (MgO) , phosphate (P2O5) , and potassium (K2O ) and pH. For mineral analy¬ sis, samples were taken two different times and the results averaged. Results Seedling — Seed Source Study: It can be seen from Table I that the pro¬ portion of the parent seed sources is strongly reflected in the seedling population. Apparently in this non-old field situation, Virginia pine is as likely to successfully invade as loblolly pine. Vegetation Analysis: Sampling data are presented in Table II. Relative dominance for all trees > 1" dbh, relative density of overstory trees (> 6" dbh), relative density for understory (> 1" dbh, < 6" dbh) trees, and stems per hectare for each stratum are recorded for each stand. Stands 3, 4, 7, and 8 are clearly dominated by loblolly pine. Stands 1, 2, 5, and 6, how¬ ever, are a fairly even mixture of the two species. This verifies the reported history/ compos ition relationship: abandoned fields allowed develop¬ ment of nearly pure loblolly stands, whereas only cut-over areas allowed mixed stands to develop. The mixed (cut-over) stands have a greater diver¬ sity of hardwood species present than the loblolly (old- field) stands. This is because, stump sprouts, and relict seedlings and saplings of hardwoods would be left in cut-over areas, but be absent from abandoned fields. Despite greater abundance in cut-over areas, none of the hardwood species seemed associated more with one type of pine stand than another. Soil Moisture: Table III shews results of average percent soil moisture readings for spring (May- June) , summer (Aug. -Sept. ) , and fall (Oct. -Nov. ) for each of the 8 stands. Three of the lob lolly- dominated stands are consistently more moist than the four mixed stands, a difference which might be critical in very dry summers like that of 1976. However, cne of the loblolly stands is just as dry as the mixed stands. Soil Texture, pH, and Mineral Content: Table III gives results of the soil analysis . All 4 loblolly stands have slightly higher pH than all mixed stands and two loblolly stands have higher K^O than any mixed stands, but the other two did not. No other patterns are discernible between the two groups. Surprisingly, the driest stands in each group are also the least sandy in their group, which is the reverse of the conventional expectation. Discussion Soil fertility seemed a reasonable possibility as a controller of distribution in these two pines, even though it was previous land use that correlated with their abundances. The cut-over areas obviously had been allowed to grow into pine forest that was old enough to be harvested before the current generation of pines was established. As agricultural land use was declined, less fertile areas would probably have been abandoned first, so natural fertility might be higher in the most recently abandoned (old field) stands. Further, artificial fertilization should DISTRIBUTION OF VIRGINIA PINE 83 Table II. Vegetational composition of studied stands. Only major species are included. Stand Type Cut-Over Stand No. 1 (Longhill Rel. Rel. Dorn. Dens. > 6" 1) Rel. Dens, dbh 1-6" 2 (Longhill 2) Rel. Rel. Dorn. Dens. > 6" dbh Rel. Dens. 1-6" 6 (Hiden Estate) Rel. Rel. Rel. Dan. Dens . Dens . > 6" dbh 1-6" 5 (Jolly Pend) Rel. Rel. Dorn. Dens. > 6"dbh Rel. Dens. 1-6" Pinus taeda 41.6 45.2 8.5 27.3 41.5 5.1 15.7 50.0 3.3 20.4 25.0 18.9 Pinus virginiana 34.5 46.8 3.4 31.8 31.0 - 37.3 33.1 29.7 68.5 25.0 75.4 Liquidanbar styraciflua 3. 11 - 1.7 - 3.51 - 17.6 16.6 19.8 - - 1.9 Liriodendron tulipifera 7.7 4.8 5.1 - - 2.6 2.0 - 3.3 - - - Nyssa sylvatica 1.5 - 20.3 4.5 - 41.0 - - - 1.8 25.0 - Comus florida - - 10.1 - - - - - 5.5 - - - Prunus serotina - - 1.7 - - - - - - - - - Quercus falcata 1.5 1.6 - 2.3 3.5 2.6 11.8 - 11.0 3.9 25.0 - Ilex opaca 3.1 - 13.6 - - - - - 2.2 - - - Oxydendron arboreum 1.5 - 10.3 - - - 3.9 - - - - - Quercus alba - - 5.1 25.0 20.6 18.0 - - 1.1 1.8 - - Carya tccnentosa 1.5 1.6 8.5 4.5 - 12.8 - - - - - - Carya pallida - - - 4.5 - 12.8 - - - - - - Diospyros virginiana " - - " - - - - - " " - Stems /'ha 684 2611 480 2583 296 4483 312 4141 Stand Type Old Field Stand No. 3 (Centerville L) Rel. Rel. Rel. Dan. Dens. Dens. > 6" dbh 1-6" 8 (College Woods) Rel. Rel. Rel. Dan. Dens. Dens. > 6"dbh 1-6" 4 (Centerville R. ) Rel. Rel. Rel. Dan. Dens. Dens. > 6"dbh 1-6" 7 (Waller Mill) Rel. Rel. Rel. Dan. Dens. Dens. > 6" dbh 1-6" Pinus taeda 100 100 75.0 99.3 98.9 42.4 94.5 95.9 34.4 93.9 93.8 20.8 Pinus virginiana - - 3.6 - - - 5.5 4.2 2.1 0.8 2.1 2.1 Liquidambar styraciflua - - 10.7 - - 7.6 - - 18.8 - - 20.8 Liriodendron tulipifera - - 3.6 0.7 1.1 28.3 - - 6.3 - - 4.2 Nyssa sylvatica - - - - - 2.2 - - - - - 2.1 Comus florida - - - - - 8.7 - - 12.5 - - 4.2 Prunus serotina - - - - - 1.1 - - 4.2 1.5 2.1 12.5 Quercias falcata - - - - - - - - 2.1 - - - Ilex opaca - - - - - - - - - - - 8.3 Oxydendron arboreum - - - - - - - - 6.3 - - - Quercus alba - - - - - - - - - - - - Carya tomentosa - - - - - - - - - - - - Carya pallida - - - - - - - - 2.1 - - - Diospyros virginiana - - - - - - - - - - - 14.6 Stems /ha 824 1647 1328 793 776 1069 781 518 1. Because dominance and density are measured separately in the conbined Bitter lich/circular quadrat method, species may be encountered only in one column or the other. 84 THE VIRGINIA JOURNAL OF SCIENCE LO CO CO CO LO oo CO LO O' O' !— 1 CO 1 — 1 CO 1— 1 O' NO LO co CO -cf LO 00 o on <1" ON CO rH CNl CO nO - 4-1 CO p*_. T3 - i-i cd -h 60 4-1 4J co 0 o 6-5 CO £ •§ T— 1 s r—l •H cd i — 1 •H CO CO CJ o CD 6-5 S'S 6-5 DISTRIBUTION OF VIRGINIA PINE 85 affect the mineral content of old fields so that at least in the earlier years after abandonment, when pine seedlings are being established, they ! might be more fertile than a cut-over forest stand long removed from i cultivation. While Oosting (1942) noted that "chance of seeding" determined to a large extent the local distribution of species in pine ! stands, with both Virginia and loblolly pine exhibiting equal vigor wherever ! they start, Oosting (1942) did note a certain degree of differential distribu¬ tion: loblolly is most often found on the "best" sites, and Virginia pine on the "poorest" sites. According to Collingwood (1938) Virginia pine will indeed invade abandoned fields, but persists only on the "more sterile", sand and clay areas of its range". Fowells (1965) also mentions the superior agressiveness of Virginia pine on poor sites. The species would thrive on moist, fertile soils, but as Collingwood (1938) stated, competition from other species here prevented survival of pure stands, leaving only scattered Virginia pines. It can be inferred that, where seed source is adequate at a given site, Virginia pine will be most likely to successfully invade under poor conditions, where it is a relatively better competitor than species less tolerant of poor soil. Not all data support this scenario. Not only did McQuilken (1940) assess site factors as minor in their influence on local distribution of these twD species, but Fowells and Kraus s (1959) reported no difference between these two species in nitrogen and phosphorus requirements. The results of our study offer no evidence to refute these latter findings; our data show that soil nutrients and soil texture are similar in both stand types. McQuilkin (1940) indicated that moisture was the major factor other than seed source in establishment, and Hosner & Graney (1970) suggested that further work should be done to determine if the twro species have different responses to soil moisture levels. It might be expected that low moisture sites would be among the first to be abandoned by farmers, and therefore the cut-over sites would be drier sites. Three of the old field stands were indeed moister than the four cut-over sites, and this might be interpreted as consistent with Fowells' description of Virginia pine as occurring on moderately well-drained to well-drained soils, with loblolly more important on poorly drained sites. It would also fit well with the idea that loblolly has a competitive advantage on wetter sites, whereas on drier sites it is less aggressive and Virginia pine is able to assume co-dominance. However, the one drier loblolly-dominated old field stand casts doubt on the significance of moisture differences in this study, since it is as dry as the cut-over sites. Hosner and Graney (1970) have shown that no significant difference of growth of seedlings occurs between Virginia and loblolly pine on either abandoned field sites or climax oak -hickory stands, but they worked only whth transplanted seedlings and their conclusion is not necessarily pertinent to our study. Their study did not encompass germination, establishment, or older stages of the two species, which in our case have all taken place under natural conditions. Further work needs to be done to determine whether loblolly gains an advantage in old fields at the time of germination or later. An explanation is still needed as to why Virginia pine is better able to compete on cut-over areas. Soil fertility cannot be completely be ruled out as the factor that tips the competitive balance, since fertility of old field just after abandonment might indeed be higher than 20-30 yrs after abandonment. While there is no convincing evidence in our data that soil moisture is crucial to the competitive balance, some interaction of moisture and other factors may be involved. Is it possible that litter on the timbered sites (which would not be on an old field) diminishes the success of loblolly reproduction but not that of Virginia pine, allowing the latter to gain a foothold? Even if it is past land use, rather than seme intrinsic factor that determines the competitive balance, the physical or biotic differences caused by the land use and altering the competitive balance need to be discerned. Further work is needed to find answers to these questions. 86 THE VIRGINIA JOURNAL OF SCIENCE Literature Cited Bowers, Lillian. 1976. Hew to judge your soil's texture. Flower and Garden, Septerrber 1976: 35-36. Collingwood, G. H. 1938. Virginia pine (Pinus virginiana Miller) . Amer. For. 44: 416-420. Fowells, H. A. 1965. Silvics of Forest Trees of the United States. Ag. Handbook No. 271. U.S.D.A. _ . and R. W. Krauss. 1959. The inorganic nutrition of loblolly pine and Virginia pine with special reference to nitrogen and phosphorous. For. Science 5: 95-112. Hosner, John F. , and David L. Graney. 1970. The relative growth of three forest tree species on soils associated with different successional stages in Virginia. Amer. Midland Naturalist 84 (2) : 418-427. McQuilkin, W.E. 1940. Natural establishment of pine in abandoned fields in the Piedmont Plateau region. Ecology 21: 135-47. Monette, R. , and S. Ware, 1983. Early Forest Succession in the southeastern Virginia Coastal Plain. Bull. Torrey Botan, Club 110: 80-86. Oosting, H. J. 1942. An ecological analysis of the plant communities of Piedmont, North Carolina. Amer. Midland Naturalist 28: 1-126. Virginia Journal of Science Volume 34, Number 2 Summer 1983 A Rapid and Convenient Procedure for the Synthesis of Radiolabeled Di- and Tripeptides Lloyd Wolfinbarger and John Castellano Department of Biological Sciences Old Dominion University Norfolk, Virginia 23508-8503 Abstract This paper describes a procedure for the rapid synthesis of radiolabeled di-and tri-peptides. The technique makes use of a water soluble coupling reagent and catalytic dehydrogenation for deblocking the synthetic product. The radiolabeled peptide may be obtained in high yield and purity. INTRODUCTION The field of peptide synthesis has expanded to such an extent over the past decade that the novice who needs only to synthesize the occasional peptide quickly becomes lost in the deluge of possible meth¬ odologies to be followed. Fortunately, the availability of a wide assort¬ ment of small peptides from commercial sources such as Sigma Chemical Co. (St. Louis, Mo.) has made it possible to buy that occasional peptide unless the researcher needs it to be radiolabeled or modified in some way. Several years ago the authors found it necessary to use small radiolabeled peptides in a research project and ware faced with the necessity to synthesize a small number of peptides on a limited budget. This paper is the result of several months of searching for a simple and reliable synthetic procedure for preparing radiolabeled peptides. Much of the individual methodologies have been published elsewhere by others and the value of the procedure presented here lies in the appli¬ cation of such methodologies in the combination described. The readers are referred to the book "Peptides: Structure and Biological Function" by E. Gross and J. Meinenhofer (Pierce Chemical Company, Rockford, Ill.) for an extensive review of peptide synthetic procedures. The synthetic procedure described in this manuscript may be perform¬ ed in less than one day and will give yields (up to 80%) of a product which may be used without further purification. The procedure utilizes a water soluble coupling reagent and protecting groups which may be simul¬ taneously removed within 2 to 5 minutes. Materials and Methods For convenience we have chosen to use synthesis of glycl-L-leucine and L-leucyl-L-pheny lalan ine for description of the procedure. The use of amino acids with additional reactive groups would necessitate the use of alternate methodologies. The carbobenzoxy (CBZ) protected amino acids (or peptides) and amino acid benzyl esters may be purchased from Sigma. Chemical Company. Radiolabeled amino acids my be purchased from Amersham Radiochemicals (Arlington Heights, IL) or a similar radiochemical company. If you purchase a radiolabeled amino acid you my find it necessary to con- This paper is reprinted here because through editorial error portions of the manuscript were omitted when it was published previously. 88 THE VIRGINIA JOURNAL OF SCIENCE struct the benzyl ester derivative yourself (see later methods) . The coupling reagent EDC (l-ethyl-3- (3-dimethylaminopropyl car- bodiimide) and deblocking reagent, palladium black, may be purchased from Sigma Chemical Company. SYNTHESIS OF BENZYL ESTER DERIVATIVES: The procedure used for the synthesis of the benzyl ester is essen¬ tially that of Zervas, Winitz and Green stein (1957) . The benzyl ester derivatives of selected amino acids are synthesized by mixing the labeled amino acid (50 yCi, 165irCi/imole) with a known amount of non¬ radioactive amino acid (20 nmoles) and adding this mixture to an equal amount (20 nmoles) of p-toluensulfonic acid. Benzyl alcohol (80 mis) and benzene (40 mis) are added and the entire solution refluxed for 8 hours in a Dean & Stark distillation apparatus with the temperature of the reaction never exceeding 80°C. Upon corrpletion of the reaction the light yellow solution is cooled to room teirperature , 500 mis of diethy- lether added and the entire solution cooled to 0° - 2°C for 15 hours to precipitate the p-toluensulfonate amino acid benzyl ester. The melt¬ ing point of the amino acid benzyl ester p-toluensulfonate is checked against the published value (158. 5-16 0°C for leucine benzyl ester) (Zervas, Winitz, Greenstein, 1957). The solution is filtered and the precipitate added to chloroform (30 mis) at 0°C with constant stirring. Triethylamine is then added dropwise until a clear solution results. Ether (400 mis) is added to precipitate the triethyl ammonium p-toluen¬ sulfonate and the etheral solution supernatant concentrated in vacuo to approximately 2 mis and saturated with HCL gas to precipitate the amino acid benzyl ester salt. The purity of the product is determined by melting point analysis, using a Hoover capillary point melting apparatus, and thin layer chro¬ matography. SYNTHESIS OF THE PEPTIDE: ri4 1 The /_ Cj - amino acid benzyl ester was used without further puri¬ fication by dissolving (10 nmoles) in methylene chloride or tetrahy- drofuran (5 mis) containing freshly distilled triethylamine (20 yl) which is used to neutralized the associated HC1. The amount of trie¬ thylamine used in any synthesis is dependent upon the amount of benzyl ester used, such that equivalents of each are reacted. An equimolar amount of carbobenzoxy-amino acid (10 mmoles) and l-ethyl-3- (3 dime- thylamino propyl) carbodiimide (10 mmoles) are added and the reaction mixture stirred overnight at room temperature. The reaction mixture is then washed three times with 25 mis of water to remove the water soluble N-acyl urea which is a by-product of the reaction and unre¬ acted CBZ amino acid, and three times with 0.1 N HC1 (25 mis) to re¬ move the unreacted benzyl ester. Evaporation of the imythylene chloride leaves the yellow oil residue of the carbobenzoxy and benzyl ester blocked peptide (CBZ-L-amino acid - L - [14cJ amino acid benzyl ester. ) REMOVAL OF PROTECTING GROUPS: The simultaneous removal of both the carbobenzoxyl and benzyl ester protecting groups is accomplished by dissolving the product in 3-5 mis methanol/ formic acid (4.4%) and passing the solution over palladium black (50-100 mg) packed in a Pasteur pipette. ElAmin, et al., (1979) reported the rapid removal of such protecting groups by catalytic transfer hydrogenation. Formic acid was shewn to be a very effective hydrogen donor for the rapid removal of these peptide pro¬ tecting groups and a good solvent component, providing complete sol¬ ubilization of most peptides and peptide derivatives. RADIOLABELING OF DI- AND TRIPEPIDES 89 1000- 900- 800- LLJ ^ 700- 1 600- 500- 400- 300- 200- 100- 1 5 (LEU-BENZYL ESTER) Q (LEU) /' To 15 20 25 30 35 40 TLC SECTION Fig. 1. Thin layer chromatography of synthesized L-[ 14C] leucine benzyl ester ( 0 - 0 ) and the product formed following passage of the benzyl ester over the palladium black column ( 0 - 0 ). The oval symbols above each peak of radioactivity indicates the position on the TLC sheet where authentic L-leucine and L-leucine benzyl ester migrated. See Materials and Methods for run conditions. 1200-1 1000- 800- £ 600- 400- 200- cd cr 1 5 -i - 1 - — i - 1 - 1 - 1 - n 10 15 20 25 30 35 40 TLC SECTION Fig. 2. Thin layer chromatography of synthesized [,4C]-glycyl-L-leucine ( 0 - 0 ) and the radiolabeled product remaining after acid hydrolysis ( 0 - 0 ). The oval symbols above each peak or along the top of the figure indicates the position on the TLC sheet where authentic glycine, glycyl-L-leucine, and L-leucine migrated. 90 THE VIRGINIA JOURNAL OF SCIENCE THIN LAYER CHROMATOGRAPHY: Purity of all benzyl esters and peptides synthesized was confirmed by thin layer chrorratography . To test the identity of all synthetic products, cellulose thin layer chromatography sheets (Eastman) were spotted with 10 yl aliquots of the appropriate solutions as well as known peptides and amino acids, developed in an ethanol: acetic acid: water (65:1:34; v/v/v) solvent, dried and sprayed with ninhydrin (0.4% solution in acetone) . In addition the peptides were hydrolyzed under vacuum in 6 N HC1 at 100°C for 18 hours in sealed tubes. The hydrolysis products were dried under vacuum over KOH, 0.1 ml of water added, 10 yl spotted on the TDC sheet, and thin layer chromatography performed using the above procedure. TNBS ASSAY OF FREE AMINO GROUPS: Deblocking of CBZ -protected amino groups on peptides was followed by use of the trinitrobenzenesulfonic acid (USES) assay. Aliquots of formic acid/methanol solutions of CBZ - blocked peptides and amino acids prior to and following passage over the palladium black column were taken to dryness, redissolved in 1 ml of buffer (0.1 M 2 -dime thy lamino- ethanol, pH 9.0) and 1 ml of 0.1% TNBS (in water) added. The TNBS was permitted to react for 30 minutes at roam temperature prior to addition of 1 ml of 10% sodium dodecyl sulphate (SDS) and 0.5 ml of 1 N HC1. Absorbance was measured at 340 ran using a known concentration of pep¬ tide for calculation of concentration of deblocked peptide. RESULTS Synthesis of benzyl ester: r 14I Synthesis of the L- [C J - leucine benzyl ester was essentially as described in the materials and methods. Purity of the product was determined using thin layer chroim tography and figure 1 reveals that the benzylation reaction was essentially 100% complete with no radio¬ activity remaining at the position (Rf value of 0.84) of leucine. Catalytic deblocking of L (14Cj leucine benzyl ester using palladium black and methanol/formic acid resulted in complete conversion to L- [14J - leucine. The extent of the reaction was determined by thin layer chromatography and scintillation counting and essentially 100% of the L- p-^Cj - leucine benzyl ester was converted to L- ^ cj - leucine with¬ in 5 minutes (fig. 1) . Peptide synthesis: Tl4 1 Once purity of the L- L Cj - leucine benzyl ester had been estab¬ lished it could be used, without further purification. Synthesis of the peptide and deblocking as described in the materials and methods yields peptide in 70% to 85% yields as calculated from the recoverable radio¬ activity. Thin layer chromatographic analysis of an aliquot of the pro¬ duct revealed a single peak of radioactivity at an R^ corresponding to the peptide synthesized (figs. 2 and 3) . Acid hydrolysis (overnight at 100°C in 6 N HC1 in a vacuum sealed tube) of each product revealed a single spot of radioactivity on TIC plates which corresponded to free leucine (figs. 2 & 3) . Spraying identical plates with ninhydrin revealed two ninhydrin positive spots corresponding to either leucine and phenylala¬ nine or leucine and glycine depending on the peptide hydrolyzed (data not shown) . Time Course of Deblocking: Although we routinely deblock the synthesized peptides by dissolving them in methanol/formic acid and passing them over a palladium black column we felt it might be informative to examine the rate at which de- RADIOLABELING OF DI- AND TRIPEPIDES 91 TLC SECTION I Fig. 3. Thin layer chromatography of synthesized [14C]-L-leucyl-L-phenylalanine ( 0 - 0 ) and the radiolabeled product remaining after acid hydrolysis ( 0 - 0 ). The oval symbols along the top of the figure indicates the position on the TLC sheet where authentic L-phenylalanine, L-leucyl — L-phenylalanine and L-phenylalanine migrated. TIME IN MINUTES Fig. 4. Time course of deblocking of CBZ-L-leucine by palladium black and 4.4% formic acid in methanol. 92 THE VIRGINIA JOURNAL OF SCIENCE blocking occurs. TO do this, we mixed 50 mg of palladium black in 5 mis of methanol/formic acid, added the blocked peptides (or blocked sub¬ strates, i.e. CBZ - glycine, etc) and removed aliquots with time. Each aliquot was taken to dryness and analyzed for free amino groups using the TUBS assay described in the materials and methods. Figure 4 shows the appearance of free amino groups (increased optical density at 340 ran) with time. The deblocking of CBZ-L-leucine was essentially 100% com¬ plete within 2 minutes (fig. 4) . DISCUSSION The synthetic procedure described in this paper is not the only or even the best procedure for the synthesis of small radiolabeled peptides. Howsver, the various techniques and reagents employed should facilitate the synthesis of such peptides by researchers who do not wish to become extensively involved in peptide synthesis. Ml reagents are commercially available and each step in the procedure may be undertaken using equip¬ ment available in most laboratories. The reagent DCCD (dicyclohexylcarbodiimide) typically employed in this reaction yields a water insoluble dicyclohexylurea product which is difficult to remove from the reaction mixture. The EDC urea deriv¬ ative is soluble in water and is easily removed along with unreacted substrates by the initial washings of the hydrophobic peptide product. The use of carbobenzoxy and benzyl ester derivatives provides end group blocked peptide products which may be rapidly and completely de¬ blocked by palladium black. ElAmin, et al. (1979) first reported the rapid removal of these protecting groups by catalytic transfer hydro¬ genation. The formic acid was shown to be a very effective transfer hydro- donor as wsll a good solvent for most peptides and peptide derivatives. The concentration of formic acid may be varied, but 4.4% proved to be most effective in deblocking a broad spectrum of protected substrates. ACKNOWLEDGEMENTS Portions of this research were supported through a grant to L. Vfolfinbarger by the Virginia Academy of Sciences. LITERATURE CITED 1. ElAmin, B. , G. M. Anan tharamaiah , G. P. Royer, and G. E. Means. 1979. Removal of Benzyl-type protecting groups from peptides by catalytic transfer hydrogenation with formic acid. J. Org. Chem. 44: 3442 - 3444. 2. Zervas, L. , M. Winitz, and J. P. Greenstein, 1957. Studies on arginine peptides I. Intermediates in the synthesis N- terminal and C-terminal arginine peptides. J. Org. Chem. 22:1515-1521. Errata: Bell, Bovard and Stratton The paper "Changes in Balance and Hand-Eye Coordination in I Educable Mentally Retarded AdulLs as a Result of an Eight-Week | Swimming Program" by G. H. Bell, Ken Bovard, and R. K, Stratton (V.J.S. 33 (2): 7-10; August 1982), contains an error in the sta- | tiscal analyses shown in table 2. The proper error term for sex I is subject within sex, not the error mean square, as originally E presented. Following this change in the analysis, the probability I values, P>F, for the statistical significance of sex, both corrected I and former, are: for OFFBAL, reduced to .07 from .0001; for TOB, I reduced to 0.62 from 0.27; and, for TTC, reduced to 0.27 from 0.046. I This has the net effect of reducing the relative importance (statis- i; tical significance) of sex from very large to borderline, from pos- | sible to doubtful, and from borderline to possible, respectively. I Clearly, sex differences cannot be presumed to be real. Repetition 1 and refinement of this work are needed to clarify this question. The authors wish to acknowledge Dr. Klaus Hinkelmann, Head, Statistics Department, Virginia Tech for identifying and correcting the error cited above. 94 Announcement: Research Grants Small Project Grants Members of the Virginia Academy of Science May apply for small project grants to support their research. These grants are usually in the amount of $500 or less and are available to support any field of research* There are two deadlines for receipt of applica¬ tions, March 15 and October 15, each year. For application forms and further information, contact: Dr. William L. Banks, Jr. Co-Director MCV-VCU Cancer Center Box 37, MCV Richmond, VA 23298 OFFICIAL PI BLICATION OF THE VIRGINIA ACADEMY OF SCIENCE ... - ' ‘ ■ u : ?. ,r> : THE VIRGINIA JOURNAL OF SCIENCE EDITOR: Stewart Ware Dept, of Biology College of William and Mary Williamsburg, Va. 23185 BUSINESS MANAGER: N. J. Fashing Dept, of Biology College of William and Mary Williamsburg, Va. 23185 ©Copyright, 1983 by the Virginia Academy of Science, The Virginia Journal of Science (ISSN 0042-658X) is published four times a year (Spring, Summer, Fall, Winter) by the Virginia Academy of Science, Department of Biology, University of Richmond, Richmond, Va. 23173. Second Class Postage paid at Richmond, Virginia and additional entry at Harrisonburg, Virginia 22801. The Virginia Academy of Science and the Editors of the Virginia Journal of Science assume no responsibility for statements or opinions advanced by contributors. Subscription rates for 1983: $27.50 per year, U.S.A.; $35.00 per year, other countries. All foreign remittances must be made at par U.S. dollars or their foreign equivalent. Back issues are available for $7.50 per issue postpaid. Changes of address, including both new and old zip codes, should be send promptly upon moving to the following address. POSTMASTER: Send address changes to Blanton M. Bruner, Executive Secretary- Treasurer, Virginia Academy of Science, Department of Biology, University of Richmond, Richmond, Va. 23173. All correspondence relating to remittances, advertising (rates will be sent upon request), subscriptions, missing issues, and other business affairs should be addressed to Business Manager. Instructions to Authors All manuscripts and correspondence about them should be addressed to the Editor. The Virginia Journal of Science welcomes for consideration original articles and short notes in the various disciplines of engineering and science. Cross-discriplinary papers dealing with advancements in science and technology and the impact of these on man and society are particularly welcome. Submission of an article implies that the article has not been published elsewhere while under consideration by the Journal. The original and two copies of each manuscript and of all figures therein are required. Authors should submit names of three potential reviewers. All manuscripts must be double-spaced through¬ out. The title, author’s name, affiliation and address should be placed on a covering page. An abstract (on a separate sheet) summarizing the text, particularly the results and conclusions, is required. After revision and final acceptance of an article, the author will be asked to furnish an error-free, camera-ready copy of the manuscript typewritten single-spaced with a black carbon ribbon on white bond paper. The text should follow the general format used by professional journals in the author’s disci¬ pline. Literature cited in the text should follow the name-and-year format: Fujishima and Honda (1972), or Spry (1969), or Guliday (1971). On the Literature Cited page at the end of the article each reference should include author(s), year, title of article, title of journal (using standard abbrevia¬ tions), volume number, and first and last page of the article. For a book, include author(s), year title, pages or number of pages, and publisher and city of publication. Examples: Fujishima, A., and Honda, K. 1972. Electrochemical Photolysis of Water at a Semiconductor Electrode. Nature 238: 37-38. Spry, A. 1969. Metamorphic Textures. Pergamon Press, New York. 350 pp. Guliday, John E. 1971. Pleistocene History of the Appalachian Mammal Fauna. In Distributional History of the Southern Appalachians, Part III. Vertebrates (Perry C. Holt, ed.). pp. 223-262. VPI & SU, Blacksburg, Va. Each figure and table should be mentioned specifically in the text, with all figure numbers and legends typed consecutively on separate sheet, and the figures identified by author’s name and figure number in pencil on the back. Table numbers and legends should be included as part of the table. Authors will be allowed 15 printed pages (including figures) free, but payment of $25 per page will be charged for the 16th and subsequent pages. Survey Form YES PLEASE REPLY! For the past five years the Virginia Journal of Science has been required by the Council of the Virginia Academy of Science to run itself on an allotment from the Academy of no more than $10,000 per year. Since these five years have included some of the worst inflation in American history, keeping within this budget has been possible only by drastic changes in format which reduced printing costs. The Editor has received numerous complaints about the much decreased attractiveness of the Journal and about the abandonment of typesetting. This survey is an attempt to gauge the degree of unhappiness about the changes, and explore possible future courses. Please tear out this page and send it to the Journal's Business Manager, Dr. Norman Fashing, Department of Biology, College of William and Mary, Williamsburg, Va . 23185, with your answers and comments marked as requested below. SURVEY OF JOURNAL FORMAT PREFERENCES 1. Have you ever submitted a manuscript to the Virginia Journal of Science? 2. Have you submitted a manuscript to the Journal since it switched to photographic reproduction of the typed manuscript? 3. Would you consider submitting a manuscript to the Journal given its present (photographed manuscript) format? 4. Would you consider submitting a manuscript to the Journal if it returned to typeset style format? 5. Even if you would not submit manuscripts, would you favor the Academy's funding a return to typeset style printing of the Journal? 6. Would you favor such a return if it meant an increase in annual dues? 7. If the Journal cannot return to typeset style printing, would you favor elimination of all issues except the Proceedings issue? 8. If the Journal were reduced to only one (Proceedings) issue a year, would you continue your membership in the Academy? 9. Have you ever served on the Council of the Virginia Academy of Science? 10. If you have comments on this problem, please write them below, or on the back of this page. VIRGINIA JOURNAL OF SCIENCE OFFICIAL PUBLICATION OF THE VIRGINIA ACADEMY OF SCIENCE Vol.34 No. 3 Fall 1983 TABLE OF CONTENTS SURVEY FORM . ABSTRACTS OF PAPERS, Sixty-first Annual Meeting of the Virginia Academy of Science, May 17-20, 1983, George Mason University, Fairfax Aeronautical and Aerospace Science . 95 Agricultural Sciences . 98 Astronomy, Mathematics, and Physics . 110 Biology . 115 Botany . 138 Chemistry . 145 Education . 158 Engineering . 162 Environmental Science . 162 Geology . 168 Materials Science . 177 Medical Sciences . 182 Microbiology . 193 Psychology . 196 Statistics . 201 Actual date of publication: November 1 1, 1983 Abstracts of Papers Presented at the Sixty-first Annual Meeting, Virginia Academy of Science May 17-20, 1983, George Mason University, Fairfax Aeronautical and Aerospace Science Section STEADY HYPERSONIC ROTATING FLOW PAST A SLENDER BODY. R. J. Barbarsky* and Houston G. Wood, III, Dept, of Mech. and Aerosp. Engrg., Univ. of Va. , Charlottesville, VA 22901. The steady, hypersonic, rotating flow of a perfect gas past a slender, stationary body represents a problem of consider¬ able interest from a basic fluid mechanics viewpoint. The object of the proposed research is to use hypersonic flow theory to develop a semianalytical model of the flow by considering the inviscid solution matched to a boundary- layer type viscous solution. The Newtonian approximation for inviscid flow is made in the course of the development. The inviscid problem appears to be amenable to similarity methods developed for nonlinear partial differential equations, while an asymptotic solution of the Prandtl boundary layer equations will be sought for the regions of "strong" and "weak" interaction of the shock and the viscous layer. Finally, conditions may be derived which permit the details of the flow in the neighborhood of the body to be replaced by a boundary condition to be imposed on the external flow field as predicted by Onsager’s Pancake model. VIDEO MODEL DEFORMATION SYSTEM FOR THE NATIONAL TRANSONIC FACILITY. A. W. Burner* , W. L. Snow*, and W. K. Goad*, NASA, LaRC, Hampton, Va. 23665. The National Transonic Facility through a combination of cryogenic temperatures and high pressure will subject models to heavier aerodynamic loads than are usually experienced in existing transonic facilities. Furthermore, the unique characteristics of this tunnel will allow independent variation of dynamic pressure while holding other critical aerodynamic variables fixed. Non-intrusive methods to measure model deformation thus become an important instrumentation consideration. Photogrammetry has been chosen as a suitable technique to determine deformation over an entire wing without scanning or requiring multiple single-point systems. Limited tunnel access dictated the selection of high resolution video instead of conventional film cameras to record data. The CCTV based measurement system to be used in the NTF in the Fall of this year will be described. AERODYNAMIC INTERFERENCE CALCULATIONS FOR AN AIRCRAFT WITH A PROPFAN. B. Chandrasekaran* and Sudhir C. Mehrotra, Vigyan Research Associates, Inc., 28 Research Drive, Hampton, VA 23666. A subsonic-flow panel code has been modified to handle the effects of a propeller wake. The effects of the propeller were modelled by a system of ring vortices of constant strength. Principles based on the blade element theory and the momentum theory were used to evaluate the swirl velocity and the pressure increase, across the propeller. Experiments are defined and model details are given. The experimental results are compared with the theory. The discrepancies between the calculated and measured quantities are analysed. Improvements to enhance the accuracy of the theoretical prediction are indicated. 96 THE VIRGINIA JOURNAL OF SCIENCE DETERMINING REYNOLDS NUMBER EFFECTS ON FLUTTER. Stanley R. Cole,* NASA Langley Research Center, Hampton, V A 23665. The effects of high Reynolds number on aeroelastic phenomenon are being studied to determine the need for aeroelastic wind-tunnel testing in the National Transonic Facility at NASA Langley Research Center when it becomes operational. The present study is an attempt to determine the effects of Reynolds number on flutter. A model has been designed for testing in the 0.3-m Transonic Cryogenic Tunnel (TCT). The 0.3-m TCT is also capable of obtaining high Reynolds numbers. The model consists of a relatively rigid wing supported by a flexible shaft. A prelimi¬ nary test of a scaled model has been conducted in the Transonic Dynamics Tunnel (TDT) to determine the flutter characteristics of the model before attempting the 0.3-m TCT test. Analytical design of the model, results of the TDT flutter experiment, and plans for the 0.3-m TCT test will be discussed. Cryogenic flutter testing procedures, including subcritical response tech¬ niques for predicting flutter onset, will also be discussed. APPLICATION OF COATED FIBER STUDIES TO SILICON-CARBIDE/TITANIUM COMPOSITES. L. L. Drewry, W. D. Brewer, and J. Unnam. NASA, Langley Research Center, Hampton, Va. 23665 A relatively new silicon carbide fiber (designated SCS-2) has shown potential as reinforcement in titanium matrix composites. As with previous titanium composites, however, the SCS-2/T i system is subject to property degrad¬ ation caused by fiber-matrix interactions. An SCS-2/T i composite system, simu¬ lated by sputter-coating individual fibers with titanium, was investigated to characterize these microstructural and chemical interactions that adversely affect material performance. Because aluminum has been shown to be an effective interfacial barrier, an SCS-2/ A1 /Ti system was also investigated. The coated fibers were exposed to time-temperature environments typical of composite fabri¬ cation, tested for tensile property changes, and analyzed by scanning electron microscopy, Auger spectroscopy, and electron microprobe analysis. Measured fiber tensile properties were used to predict composite properties to compare with measured composite properties. ENVIRONMENTAL CONTROL AND LIFE SUPPORT SYSTEMS (ECLSS) TECHNOLOGY ASSESSMENT FOR MANNED SPACE STATIONS. John B. Hall, Jr. and Shelby J. Pickett*, NASA Langley Research Center, Hampton, Va. 23665. The synthesis of an Earth¬ orbiting space station to enhance man's permanent presence in space requires the assessment of ECLSS technology options from which to select the most promising subsystem candidates for space station development. This presentation gives a description of a computer-aided technology assessment program developed at the NASA Langley Research Center to aid in selecting those options which significantly reduce the crew resupply requirements for metabolic oxygen and potable water. The data base and methodology are presented which provide the basis for the assessments as functions of crew size, mission duration, resupply interval, and life cycle costs. APPLICATION OF A LASER RESONANCE DOPPLER VELOCIMETER AT THE NASA LANGLEY RESEARCH CENTER 3-INCH HYPERSONIC HELIUM TUNNEL. John C. Hoppe, Instrument Res. Div., Langley Res. Ctr., Hampton, Va. 23665. Laser-induced fluorescence can be used to provide measurements of mean velocity high speed flowing gases. A system to accomplish such measurements has been implemented at the Langley Research Center's 3-inch hypersonic helium tunnel. Free stream velocities in that facility are approximately 1.5 to 2.0 km/sec. The method, developed at Princeton Univ.vG, includes seeding the flow with sodium atoms. The sodium absorbs incident visible radiation from a tunable dye laser, and emits D-line resonance fluorescence. The optical frequency of that emitted fluorescence is compared to a similar induced emission from an atomic beam source, to permit the velocity-dependent Doppler shift to be measured. Preliminary data are presented. R. B. Miles and M. Zimmerman, Final report for NASA Grant NSG 1070, Princeton Univ. Dept. Mech. Aerospace Eng., Report No. T-1465, March 1980. THE VIRGINIA JOURNAL OF SCIENCE 97 INVESTIGATION OF FAST INITIALIZATION OF SPACECRAFT BUBBLE MEMORY SYSTEMS. Karen T . Looney* and Paul J. Hayes,* NASA Langley Research Center, Hampton, VA 23665, Mail Stop 470. Bubble domain technology offers significant improvement in reliability and functionality for spacecraft onboard memory applications. In considering potential memory system organizations, minimization of power in high capacity bubble memory systems necessitates the activation of only the desired portions of the memory. In power strobing arbitrary memory segments, a capa¬ bility of fast turn-on is required. Bubble device architectures, which provide redundant loop coding in the bubble device, limit the initialization speed. Alternate initialization techniques have been investigated to overcome this design limitation. An initialization technique using a small amount of external storage has been demonstrated. This technique provides several orders of magni¬ tude improvement over the normal initialization time. QUASI-VORTEX-LATTICE METHOD FOR WINGS WITH EDGE VORTEX SEPARATION. Jenn-Louh Pao? Vigyan Research Associates, Inc., 28 Research Drive, Hampton, VA 23666. An improved vortex filament- vortex core method for predicting aerodynamic characteristics of slender wing with edge vortex separation has been developed. Semi-empirical but simple methods are used to determine the initial position of the free sheet and vortex core. Comparison with available data indicates that: (1) the present method is generally accurate in predicting the lift and induced drag coefficients but the predicted pitching moment is too positive; (2) the spanwise lifting pressure distributions estimated by the vortex core solution of the present method are significantly better than the results of Mehrotra's method to the pressure peak values for the flat delta; (3) the two core system applied to the double delta and strake wing produce overall aerodynamic characteristics which have good agreement with data except for the pitching moment; (4) the computer time for the present method is about two third of that of Mehrotra's method. RESIDUAL STRESS MEASUREMENT BY HOLE DRILLING METHOD IN ORTHOTROPIC MATERIALS USING PHOTOELASTIC COATINGS. R. Prabhakaran and C. B. Prasad , Mechanical Engineering and Mechanics, Old Dominion Univer¬ sity, Norfolk, YA 23508. The applicability of photoelastic coat¬ ings, in conjunction with the hole-drilling method, to the determi¬ nation of residual stresses in relatively thin orthotropic materials was investigated. The birefringent response on the boundary of a small hole drilled in an orthotropic material was calculated as a function of the principal stress-ratio for the particular case when the principal stress directions coincided with the material symmetry axes; the stress distribution is known from Lekhnitskii ' s solution. It is shown that the residual stresses can be determined by measuring the birefringent response at selected points on the hole-boundary and using calibration constants determined for simple specimens. STABILITY ANALYSIS OF THE COMPRESSIBLE EKMAN LAYER. John R. Spall* and Houston C. Wood, III, Dept- of Mech. and Aerosp. Engrg., Univ. of Va,, Charlottesville, VA 22901. A linear stability analysis of a compressible Ekman boundary layer on a rotating disc is presented. The equations of motion are reduced to solving an eigenvalue problem for the growth rates. A numerical solution is obtained using the shooting method. The rotation speed at the periphery of the disc is used to define a Mach number for the gas of interest. A Reynolds number is defined using the local boundary layer thickness and geo- stmphic flow velocity. For low Mach numbers, two wave types, called A and B waves, appear at Reynolds numbers of approximately 55 and 115 which is similar to the incompressible case. As the Mach number is increased above 7 for air, a new type of wave emerges at a Reynolds number of approximately 62. Numerical studies have shown that prescribing an adiabatic boundary condition on the rotating surface tends to be destabilizing with the critical Reynolds number reduced by a factor of three in some cases. 98 THE VIRGINIA JOURNAL OF SCIENCE OXIDATION OF COMMERCIALLY PURE TITANIUM. J. Unnam and R. N. Shenoy* , Vigyan Research Associates, Inc., Hampton, VA 23666. Titanium is an excellent candi¬ date material for thermal protection systems in hyper- velocity vehicles. Oxi¬ dation and oxygen embrittlement are the primary considerations in its poten¬ tial use at temperatures above 1000°F. Air contamination studies have there¬ fore been carried out on a commercially pure titanium in the temperature range of 1100-1400°F. Total oxidation kinetics were determined by thermogravimetric method, oxide thickness was estimated by X-ray diffraction and microscopy, and oxygen depth profiles were deduced from microhardness measurements on the spe¬ cimen cross-section. Based on these data, a model for the oxidation of titan¬ ium is proposed. IMPACT PROBE RAREFACTION EFFECT MEASUREMENTS IN C02 FLOWS. David J. Yurkanin* and Sam S. Fisher, Dept, of Mech. and Aerosp. Engrg. , Univ. of Va., Charlottes¬ ville, VA 22901. The effect of flow rarefaction upon the pressure sensed by an open-ended, tube-type impact pressure sensing probe immersed in supersonic C02 flow is examined experimentally. Experiments are carried out in free- jet gas expansions into a vacuum chamber. Measurements are obtained as a function of probe Reynolds number, free-stream Mach number, probe length-to-diameter ratio, and ratio of probe to flow stagnation temperature. Results are con¬ sidered in terms of the ratio of measured-to-ideal impact pressure, p.. At sufficiently high Reynolds numbers, p. approaches unity. At sufficiently low Reynolds numbers, p. approaches its predicted free molecule flow limit (usually large compared to unity) . Observed behaviors at intermediate Reynolds numbers* are explained. Because CO,, is vibrationally frozen in these flows, the data obtained are found to be m good agreement with previous results for air and nitrogen. Flow diagnostics applications for these probes and ex¬ tensions of observed behaviors to other flow conditions are discussed. Agricultural Sciences Section POPULATION STRUCTURE AND ANGLER HARVEST OF SMALLMOUTH BASS UNDER DIFFERENT REGULATIONS IN THE NEW RIVER. Douglas J. Austen* and Donald J. Orth. Dept, of Fisheries & Wildlife Sci., Va. Polytechnic Inst. & State Univ., Blacksburg, Va. 24061. A 305 mm (12-inch) minimum length limit exists for the angler harvest of smallmouth bass (Micropterus dolomieui) in the New River from Claytor dam, Va., to the West Virginia border. West Virginia has no minimum length limit and both states have a daily creel limit of 8 smallmouth . bass per angler. Angler harvest and catch and release of smallmouth bass was investi¬ gated as well as age structure, growth, food habits, proportional stock den¬ sity, and condition. A total of 357 anglers were interviewed who had fished an average of 4.1 hours (VA) and 3.3 hours (W V) when interviewed. Virginia anglers harvested an average of 0.11 smallmouth bass per hour and released 1.54 bass per hour whereas West Virginia anglers harvest 0.61 smallmouth bass per hour and released 1.05 bass per hour. Average length of smallmouth bass harvested was 317 mm for Virginia and 242 for West Virginia. THE VIRGINIA JOURNAL OF SCIENCE 99 OVIPOSITION TIME IN JAPANESE QUAIL AND RELATED PRODUCTION TRAITS. W. L. Beane, C. L. Bish* and F. E. Robinson*, Poultry Science Dept., VPI&SU, Blacksburg, Virginia 24061. In the Japanese quail populations used in this study, eggs from the white egg producers tend to have weaker shells than those from pigmented egg producers. To determine if time of lay or time interval between ovipositions is associated with this difference, eggs from 144 birds of each egg color group were observed for a 15-day period for time of lay, time interval between consecutive eggs, egg weight, specific gravity, and weight of shell with membranes. Analyses of data, with intervals over 36 hours deleted, showed no difference in average time intervals between the white and pigmented egg groups (24.7 vs 24.6 hr) or the average time of lay (4:07 vs 4:02 pm). The following measurements were found to be significantly different between the j white and pigmented egg groups, respectively: egg weight (11.80 vs_ 11.89 g) ; J specific gravity (1.0715 vs 1.0766); shell weight (.832 vs .918 g) ; and percent i shell (7.07 vs 7.74). Interval between ovipositions was positively correlated ! with egg weight, shell weight and percent shell, whereas time of lay was j negatively correlated with these traits. However, since no differences between the two egg color groups were noted for time of lay or interval between eggs, the difference in shell quality was not due to these factors. ! USING WILLHAM'S COMPUTER PROGRAM TO TEACH GENETIC SELECTION PRACTICES IN BEEF CATTLE: TWENTY-FIVE YEARS' RESULTS IN THREE WEEKS' CLASS TIME. K. FU Bovard, An. Sci . ; G. L. Nunn, Computing Ctr. ; M. L. McGilliard, Dairy Sci.; and, G. L. Minish, An. Sci., Virginia Tech, Blacksburg 24061. In 1970, Prof. R. L. Wi Ilham, An. Sci. Dept., Iowa State Univ., Ames developed a computer program that simulated a beef cattle breeding operation. For the 1983 Winter qtr. 115 ' students in the beef prod'n class, AnSc 4040 at Va Tech, used the Will ham pro- 1 gram as a class exercise. Thirty-five herds of 50 cows and 5 sires, each, were I simulated, and assigned to teams of three students, each. Both genetic and environmental variation were simulated in five traits, including a random 5% I mortality. For each calf crop computer output included calves' pedigrees and I; performance; progeny performance ave's of sires, and dams; estimated breeding values (EBV's) and accuracy; and actual BV's. Final grades for the exercise i? were determined partly on the basis of change in herd performance levels effect- ed by selection. Computer costs were less than one dollar per student. | MODELING OF AGRICULTURAL OPERATIONS. Nelson L. Buck,* David H. Vaughan, and j Harold A. Hughes.* Dept, of Agricultural Engineering, Va. Polytechnic Inst. & I State Univ., Blacksburg, Va. 24061. The simulation language SLAM allows the physical layout of a farm and the sequence of operations in a forage harvesting jj| operation to be combined in a graphical representation called a network. Enti¬ ties representing equipment move through the network. The arrival of equipment 1 at certain points calls FORTRAN programs that calculate the transfer of materials ij and consumption of resources. The model allows different harvesting procedures j to be compared as to resource requirements and value of the harvested crop. The | model makes use of established formulas for its calculations and is verified by ! comparison of its predictions to data collected during an actual harvest. Where ';i there may be doubt about the accuracy or validity of formulas used, the sensiti- i vity of the model to those formulas is analyzed. Applications of the model in |j research include the evaluation of new techniques before expensive field trials | are run. Applications in farm management include aiding in the selection of i! machinery and suggesting improvements to operating procedures. 100 THE VIRGINIA JOURNAL OF SCIENCE USE OF OVARIAN 20-ALPHA HYDROXY STEROID DEHYDROGENASE ACTIVITY AS A MARKER FOR THE ONSET OF PARTURITION IN RATS. George E. Bunce, Eric A. Gordon, Dept, of Biochemistry, Va. Polytechnic Inst, and State Univ. , Blacksburg, Va. 24061. Ovarian 20-alpha hydroxy steroid dehydrogenase transforms progesterone, which during pregnancy maintains the uterine lining and prevents uterine contractions into an inactive hydroxylated form. Following the withdrawal of the protective influence of progesterone, other hormones can act on the uterus to induce par¬ turition. Evaluation of this ovarian enzyme shows a rise in enzymatic activity from non detectable levels about 24 hours before parturition to maximum levels at parturition. Pregnant rats fed a low Zn containing diet show a delay in delivery time as compared with control (normal Zn) diet populations. We have also seen a concomitant delay in the time at which ovarian enzymatic activity first becomes detectable in the low Zn population as compared to the control populations. Thus it appears that quantification of 20-alpha hydroxy steroid dehydrogenase activity is a reliable marker for impending parturition in rats. COMPATIBLE CUBIC VOLUME AND BASAL AREA PROJECTION EQUATIONS FOR THINNED OLD-FIELD LOBLOLLY PINE PLANTATIONS. Harold E. Burkhart and Peter T. Sprinz, Dept, of Forestry, Va. Polytechnic Inst. & State Univ., Blacksburg, VA. 24061. Compatible equations for predicting cubic volume and basal area growth were developed by simultaneously estimating the coefficients in both models. The simultaneous estimation procedure gave stable estimates for the basal area projection equation regardless of the merchantability definitions imposed in the volume projection equation. Prediction equations were fitted using the simultaneous estimation procedure with growth data from thinned loblolly pine plantations established on old-field sites in the Piedmont and Coastal Plain of Vi rginia. PREDICTION OF FOREST TYPES IN SOUTHWEST VIRGINIA USING TOPOGRAPHIC INFORMATION. Michael L. Fies*, Dept, of Fish. & Wildl. Sci. , Va. Poly tech. Inst. & State Univ., Blacksburg, Va. 24061, & R. H. Giles, Jr.*, Dept, of Fish. & Wildl. Sci., Va. Polytech. Inst. & State Univ., Blacksburg, Va. 24061. Vegetation mapping has traditionally been a tedious yet necessary task for wildlife managers. Aerial photography with subsequent ground truthing has been used most often in large forested areas. In this study, a technique was developed to predict forest cover types using topographic information. Elevations, slopes, aspects, and topographic shapes were incorporated into a dichotomous key to determine the forest type most likely to occur in a 3-acre map cell. Classification accuracy was between 75 and 80 percent. Results suggest that a computerized geographic informa¬ tion system could be used to map large areas at a low cost. MAXIMUM YIELD RESEARCH ON CORN. D. L. Hallock, F. S. Wright*, P. H. Reid and N. L. Powell*. Tidewater Res. and C.E.C., USDA ARS, VPI&SU, Suffolk, VA 23437 Machine planted corn yields averaged as high as 244 bu/a in irrigated corn grown in 15-in wide rows having a population of 35,138 plants/a in tilled soil fertilized as indicated below. Corn emerged after planting more quickly and was 8 in taller 60 days after planting in tilled than no-till plots. Yields were 21 bu/a higher in 15-in than in 30-in wide rows. Row width variance did not affect average ear weight. Average yields were similar in plots fertilized at 75% as in those fertilized at 150% of the following rate in lb/a: 508 N, 128 P, 270 K, 141 S, 67 Ca, 33 Mg, 1.5 B, 3 Mn, 2 Zn, and 0.6 Cu. Total dry matter production varied among plant populations as follows: 37,099 > 42,906 or 32 ,093 > 19 ,965. Stalk barrenness was much higher at 42,906 plants/a than for the three lower populations. Individual plots which yielded more than 250 bu/a had more uniform plant emergence and within row spacing than plots which yield¬ ed less than 200 bu/a. Highest plot yield was 286 bu/a. THE VIRGINIA JOURNAL OF SCIENCE 101 ABSOLUTE DENSITY SAMPLING FOR OVERWINTERING ADULT ALFALFA WEEVIL, HYPERA POSTICA (COLEOPTERA: CURCULIONIDAE) . Daniel J. Hilburn, Dept, of Entomol., Va. Polytechnic Inst., Blacksburg, \a. 24061, & W. A. Allen, Dept, of Entomol., Va. Polytechnic Inst., Blacksburg, Va. 24061. A new sampling device has been developed for measuring absolute densities of overwintering alfalfa weevil populations. The hand-operated device collects stubble, litter, and soil to a depth of 3 cm in a .05 m^ area. Adult alfalfa weevils and a variety of other insects can be extracted from the samples with Berlese funnels. EFFECTS OF ELECTRICAL STIMULATION ON BROILED BEEF FLAVOR AS MEASURED BY THE RBU SYSTEM. C. L. Hanson, R. F. Kelly, R. W. Young, K. P. Bovard, and P. P. Graham. Depts. of Food Science § Technology, Biochem. 6 Nutr. , and Animal Sci., Va Polytechnic Inst., Blacksburg, Va. 24061. One side of five beef carcasses of similar quality was subjected to 18 electrical pulses of 550 volts for 30 seconds. The opposite sides served as controls (NES) . Thirteenth rib steaks from both sides were cooked in a research broiler unit (RBU) and evaluated by a sensory panel. During cooking, headspace volatiles and drippings were collected, prepared and analyzed by GLC for fatty acids and aromatic compounds. Differences due to animal, treatment (ES vs NES) and test samples (volatile vs drip) were small and non-significant (P>.05). However, the stearic acid value for ES samples was 3.8% as opposed to 2% for NES samples (P>.09). Differences in sensory panel results due to ES were non¬ significant but the effect of ES on aroma and overall acceptability approached significance (P>.13). ECOLOGICAL INSIGHTS INTO EASTERN RED CEDAR (JUNIPERUS V I RG INI ANA L.) AS AN AGRICULTURAL PEST IN VIRGINIA. A.M.A. Hoi thui jzen* and T. L. Sharik, Dept. Fish, and Wild!. Sci. and Dept. Biol., VPI & SU, Blacksburg, VA 24061. Red cedar is one of the first woody species invading abandoned pastures in Virginia and is considered a serious pest by farmers. Red cedar stands can be fully stocked in six years on undisturbed, abandoned pastures. Rapid invasion of pastures is induced by endogenous dispersal of seeds by avian dispersers. Flock feeding bird species play an important role in red cedar seed dispersal and several of these species (e.g, starling) forage in active pastures. Seeds passed through birds are viable and possess a higher total germination as well as a higher germination rate than untreated depulped seeds. Red cedar seeds apparently do not accumulate in the soil as was established by sampling red cedar stands of increasing age. Burial of red cedar seeds showed that only 16% of the seeds were viable after 3 months. Thus, red cedar regeneration mainly originates from annual seed rains. EFFECT OF BREED OF SIRE AND GRANDSIRE ON THE PERFORMANCE OF THREE-BREED-CROSS COWS. Gretchen Kirsch* and Thomas J. Marlowe, Virginia Tech., Blacksburg 24061. Data were available on 315 matings from a larger study of cow types. Sire breeds of these cows were Angus (A), Shorthorn (Sh) and Simmental (Sm). Sire differences were significant for weaning wt (ww) , weaning rate and conformation and condition scores and approached significance for calf mortality (P <.07), ad j . 205d wt (P <.09) and kg calf wn/cow exposed (P < . 06 ) . Cows by Sh sires weaned more calves and more total calf weight, whereas differences between AA and Sm sired cows were small and nonsignificant. Grandsires of these cows were Angus (A) , Shorthorn (Sh) , Brown Swiss (BS) , Holstein (F) , Charolais (C) and Simmental (Sm) . Effect of breed of grandsire of cow was significant only for condition of calf at weaning. Calves with C and F grandsires were lowest and Sh calves highest with others intermediate. Differences in kg of calf weaned were nonsignificant but favored F (177 kg) and Sm (181 kg) grandsires with C (157 kg) and BS (158 kg) grandsires the lowest. 102 THE VIRGINIA JOURNAL OF SCIENCE EFFECTS OF PLANT GROWTH HORMONES ON OVIPOSITION SITE SELECTION OF LIRIOMYZA TRIFOLII (BURGESS) (DIPTERA: AGROMYZIDAE) J. J. Knodel-Montz*, S. L. Poe, and R. E. Lyons*, Dept, of Entomol., Va. Polytechnic Inst. & State University, Blacksburg, VA. 24061. The effects of three plant growth hormones (GA NAA, B9) and leaf position on oviposition preference of Liriomyza trifolii (Burgess) were measured. All hormones and water (control) were applied as foliar sprays to White Iceberg chrysanthemums. GA^ ^ (gibberellin aggravator) caused significantly greater (P=0.05) mean mine density than controls indicating that GA treatments enhanced oviposition. By contrast, B9 (gibberelin inhibitor) resulted in significantly (P=0.05) lower mean puncture densities when compared with controls. On chrysanthemums, lower leaves had significantly higher mine densities and lower punctures densities than upper leaves. Therefore, older leaves are preferred for oviposition while younger leaves are preferred for feeding. WITHIN PLANT DISTRIBUTION OF THE IMPORTED CABBAGEWORM, ARTOGEIA RAPAE (L.), ON CABBAGE. J_. A. Lasota and L. T. Kok, Dept, of Entomology, Va. Polytechnic Inst, and State Univ., Blacksburg, Va. 24061. The imported cabbageworm, Artogeia rapae (L.), is a major pest of cruciferous crops in southwest Virginia. Fre¬ quent applications of chemicals are employed to decrease economic losses to fresh market cabbage plantings. Knowledge of the spatial distribution of the imported cabbageworm within plants is useful in establishing damage threshold levels for reductions in the use of chemicals. Sampling to determine threshold levels can be facilitated by an understanding of the insect life stages and their distribution on the cabbage plant. Recognition of the niche requirements of the imported cabbageworm also aids in studies of parasitism by natural enemies, and population suppression by a naturally occurring granulosis virus. Our investigation shows that variations in oviposition, larval feeding, and pupation sites of the imported cabbageworm are related to changes in season and host phenology. Comparisons of pest population distribution on foliage of prehead versus headed plants exhibited more larvae and pupae on heads than on individual leaves. CONTINUED EVALUATION OF SALINOMYCIN AS A GROWTH PR0M0TANT FOR SWINE. M. D. Lindemann & E. T. Kornegay, Dept, of An. Sci . , VPI & SU, Blacksburg, Va. 24061. Four trials were conducted at three locations to evaluate the effects of graded (0, 27.6, 55.1, 82.7 and 110 ppm) levels of salinomycin on the rate and efficiency of growth of swine fed corn-soybean meal based diets. Data were broken into three phases: an initial period of approximately four weeks, a second four week period and, then, the remaining trial period. Pigs fed diets to which salinomycin had been added gained weight faster than control pigs during the first (P<.08), second (P<.01), third (Pc.10) and total trial ( P< . 01 ) periods. The pigs fed added salinomycin diets also gained weight more effi¬ ciently during each of the first two periods ( P< . 001 and P<.04, respectively) as well as for the total trial period ( P< . 01 ) . For the total trial period (8.8 to 96.1 kg) the average daily gain was 719, 731, 740, 749 and 743 g and feed/gain was 2.94, 2.89, 2.89, 2.82 and 2.87 for pigs fed diets with increas¬ ing levels of salinomycin. For the total trial period pigs fed the most efficacious level (82.7 ppm) had a 4.2% improvement in average daily gain and a 4% improvement in feed/gain compared to pigs fed the control diet. THE VIRGINIA JOURNAL OF SCIENCE 103 EFFECT OF THE ADDITION OF PEANUT HULLS TO DIETS FOR GROWING-FINISHING SWINE. M. D. Lindemann & E. T. Kornegay, Dept, of An. Sci . , VPI & SU, Blacksburg, V a. 24061. Two trials were conducted to evaluate peanut hulls (IFN 1-03-629) as a feedstuff for swine. Dietary treatments were 0, 7.5, 15 or 22.5% peanut hulls in a corn-soybean meal based diet. The effects on performance were similar for the growing (28.7 to 60.3 kg), finishing (60.3 to 98.7 kg) and overall trial periods. No differences were observed with respect to average daily gain (P>.40). Feed intake (P<.05) and feed/gain ( P< . 00 1 ) increased linearly as the level of peanut hulls increased in the diet. For the total trial period, average daily gain was 779, 777, 770, and 769 g, average daily feed intake was 2283, 2450, 2483 and 2578 g, while feed/gain was 2.93, 3.15, 3.23 and 3.35, respectively, for pigs fed diets containing 0, 7.5, 15 and 22.5% peanut hulls. Estimates of the contributions of gastrointestinal contents to the final weight were measured by subjecting animals to a 48 hr fast upon completion of the study. Weight loss was greater (linear, P<.004) with the incremental additions of peanut hulls to the diet (5.68, 6.73, 6.71 and 7.22 kg, respectively). ge in THE COMBINED TOXICITY OF CARBARYL, CHLOROTHALCNIL AND MALATHION TO THE COLORADO POTATO BEETLE, SQUASH SEEDS, AND TCMATO SEEDS. Louis P. Lue, C.C. Lewis, and V. E. Melchor, Dept, of Agric. and Chem. , Va. State Univ. , Petersburg, Va. 23803. We have kept 5 morphs of the Colorado potato beetle in our lab since we dis - covered them in 1979. Their first instars responsed to chemicals differently. The LD^q's of carbaryl were 22, 34, 50, 59, 68 ppm for morphs V, C, J, L, and B respectively. Malathion had LD^q's: 87. 50. 55. 40. and 105 ppm to V, C, J, L and B insects. Chlorothalonil was less toxic to this insect, and it had LD^q's: 1,500, 2,100, 1,400, 935 and 3,200 ppm to types V, C, J, L, and B. Morph C had the highest population of the five, therefore, this morph was selected for tests of combinations of these three pesticides at LD q's when carbaryl and chlorothal¬ onil were applied together concentration of their mortality increased 20-40% or up to 90%. While chlorothalonil and malathion increased mortality up to 80%, carbaryl and malathion increased killing up to 100%, and all three together performed 100% kill. Squash and tcmato seeds were grown in peat pots of soil mixture containing carbaryl, chlorthalonil and malathion at recommended rates, one tenth and a hundredth of the recommended. We found that carbaryl inhibited germination of both seeds most and malathion the next, however chlorothalonil seemed to enhance the germination. THE STRUCTURE AND FUNCTION OF THE OVIPOSITOR OF HYLOTRUPES BAJULUS (L.) (COLEOPTERA: CERAMBYCIDAE) . J. T. Mares and W. H Robinson, Dept, of Entomol., Va. Polytechnic Inst. & State Univ., Blacksburg, VA 24061. The Old House Borer, Hylotrupes bajulus (L.) (Coleoptera : Cerambycidae) (OHB) , is a serious pest of seasoned softwoods. Adult OHB females select cracks and crevices in the various softwoods, especially Pinus spp, for oviposition sites. Existing research shows that the female OHB uses her ovipositor to test the suitability of the substrate for oviposition. Examination by scanning electron and light microscopy indicates that the ovipositor consists of the modified abdominal segments VI-X. Various types of sensory receptors can be found on the ovipositor especially on the terminal segment and cerci. Mechanoreceptors are comprised of long tactile hairs and campaniform receptors. Chemoreceptors consist of multiple-dendrite hairs and pegs. Using fasted body weight, avera 703, 695. and 689 g) and feed/ga and 3.74) with increasing level daily gain decreased (P<.08) linearly (717, increased ( P< . 001 ) linearly (3.19, 3.49, 3.57 peanut hulls. 104 THE VIRGINIA JOURNAL OF SCIENCE COMPARATIVE PERFORMANCE OF THREE COW TYPES WHEN MATED TO TWO KINDS OF CROSSBRED BULLS. T. J. Marlowe, Gretchen Kirsch* and K. Nadarajah. Va Tech, Blacksburg 24061. Records on fertility, calf survival, and growth performance to weaning on 234 straightbred (SB), 487 single cross (2W) and 317 three-breed cross (3W) cow exposures at 3 locations to Angus (AA) , Limousin x Shorthorn (LS) and Maine- Anjou x Shorthorn (MS) bulls were analyzed by least squares (LS) procedures. Crossbred cows weaned more kg of calf than SB cows when mated to the same bulls. Calf losses were greatest (16.6%) and weaning rate lowest (74.7%) for the 2W cows. Adj . 205d wts were lowest for SB cows (170 vs 193 and 190 kg). Cow rank, based on kg calf wn/cow exposed, was SB (132 kg), 2W (147 kg) and 3W (162 kg). Other differences were small and generally nonsignificant. Calves sired by MS bulls were superior to LS sired calves by 31 kg/cow due solely to higher weaning rate. AA sired calves were intermediate. Because AA bulls were used only on first calf heifers they had an advantage in conception rate but the major dif¬ ference was in calf losses (1.1 vs 18.0 and 14.1%). EFFECT OF MATING TYPE ON COW AND CALF PERFORMANCE. T, J. Marlowe and K. Nadara¬ jah, Virginia Tech, Blacksburg 24061. Limousin x Shorthorn (L3) and Maine Anjou x Shorthorn (MS) bulls were mated to several F]_ cow types at 4 locations (loc) for 2 calf crops (1979, 1980). Cow types compared were Angus (A) and Hereford (H) straightbreds (PB) . Crossbred types were AH, B Swiss x H (BH) , Charolais x A (CA) , CH, Holstein x A (FA), FH, Shorthorn x H (SH) and Simmental x H (SH) . Be¬ cause cow type differed among loc, data were analyzed within loc. Number of mat¬ ings by loc were Bland (Bl) 340, Hanover (Ha) 260, Southampton (So) 293, and State Farm (SF) 558, with total matings of 621 to LS and 627 to MS bulls and 203 PB controls. Sire breed was signif only at So favoring MS by 23 kg/cow. Highly signif diff among cow type at all loc. All XB cows were superior (sup) to all PB cows at all loc except for Sm cows at Bl. FA and FH cows sup at all loc except at Bl where they tied with SH. Other XB cows were intermediate with Con¬ tinental and dairy Xs generally above British Xs. CH and SH were similar in wn wt but CH lower in weaning rate and total wt weaned. RELOCATION OF TWO IMPORTED THISTLE FEEDING INSECTS ALONG THE INTERSTATE HIGHWAY IN VIRGINIA. T. J. McAvoy* , W. Mays and L. T. Kok, Dept, of Entomology, Va. Polytechnic Inst, and State Univ. , Blacksburg, Va. 24061. Two thistle feeding weevils, Rhinocyllus conicus and Trichosirocalus horridus, were imported from Southern Europe and released in Western Virginia from 1969-76 as biological control agents of Carduus spp. thistles. jT. horridus and R. conicus are now established in 7 and 22 counties west of the Blue Ridge, respectively. In June of 1981 and 1982, releases of 500 R.. conicus and 125-150 T_. horridus were made at 21 sites from Charlottesville north to Loudon County and at 6 sites from Winchester south to Lexington along the primary highways. In 1982, _R. conicus was observed at all 27 sites except for one site in Greene County. Establish¬ ment of T. horridus was observed at 2 sites in Albermarle County and 1 site in Fauquier County. DAMAGE POTENTIAL OF CEREAL LEAF BEETLES IN VIRGINIA SMALL GRAINS. Robert M. McPherson, Dept, of Entomology, Va. Polytechnic Inst, and State Univ., Blacks¬ burg, Va. 24061. Cereal leaf beetle, Oulema melanopus (L.), population densi¬ ties were evaluated in controlled field plots in soft red winter wheat, winter barley, spring oats, and field corn during 1980-82. Populations of 1.3-1. 6 beetles/culm significantly reduced wheat yields 5.8 bu/a (17.2%). Populations of 0.6/culm reduced barley yields 2.3 bu/a below the treated plots. Densities of 7.7-11.6/culm reduced oat yields an average of 29.5 bu/a (60.9%). Field corn yields were not significantly affected by beetle feeding even though defoliation approached 40-50% during late June. More beetle eggs were observed in oats treated in early spring with 1.12 kg ai/ha of disulfoton to control aphids than in untreated plots. Similar treatments in wheat and barley did not . influence egg numbers. THE VIRGINIA JOURNAL OF SCIENCE 105 EFFECTS OF CULTIVAR, ROW PATTERN AND SEEDING RATES ON THE YIELD, VALUE AND GRADE OF VIRGINIA TYPE PEANUTS. R. Walton Mozingo and T. A. Coffel t*, Tidewater Res. and Continuing Education Ctr. , VPI&SU and USDA ARS, Suffolk, VA 23437. The effects of cultivars, row patterns and seeding rates on the yield, value and grade of Virginia- type peanuts were studied for four years at the Tidewater Research and Continuing Education Center in Suffolk, Virginia. The cultivars Flori giant and Virginia 81 Bunch were seeded at the rate of 58,080 and 87,120 seed per acre in a single and double row pattern. Virginia 81 Bunch had a higher percentage of extra large kernels, sound mature kernels, total meat and value per acre than did Florigiant. No differences were found for yield between the cultivars; however, Florigiant produced a higher percentage of fancy size pods. The only effect of row pattern was on the percentage of fancy pods and total meat where the single row pattern was higher than the double row pattern. Seeding rates had no effects on the grades. However, significantly higher yield and value per acre was recorded for the higher seeding rate. { COMPARATIVE GROWTH PATTERN OF COWS SIRED BY BRITISH AND CONTINENTAL BEEF AND | AMERICAN DAIRY BULLS AND OUT OF HEREFORD DAMS. K, Nadar a.j ah, T. J. Marlowe and D. R. Hotter, Virginia Tech, Blacksburg 24061. Growth patterns and condition (cond) scores of 92 Herefords (H) and 299 crossbred (XB) female progeny of H dams sired by two British (Br) (Angus, A, and Shorthorn, S) , two Continental il (Cont) (Charolais, C, and Simmental, S) and two American dairy (Da) breed (Brown Swiss, B, and Holstein, F) bulls were compared from birth to maturity (M) . Breed types differed signif. Both straightbred (SB) H and Br crosses (BrX) were lighter than Cont and Da Xs. BrX and SB H diff only slightly. Among XB, SH wt jl most (34 kg) at birth and AH wt least (30 kg). Wn wt of Cont X were 2.6 kg more i| than BrX and 1.7 kg more than DaX. At 30 mo Cont and Da Xs wt 41 and 23 kg more ' than BrX. DaX were intermediate. Breed type means at each age were used to fit ! Brody and Richards growth models. Asymptotic wts of ContX were highest, fol¬ lowed by DaX. BrX and HH avg less at M (487 kg) . CH cows M more slowly but grew to heavier M wt (582 kg) than SH cows (563 kg). Among the BrX, SH reached M wt well before AHXs. DaXs had almost ident M wt (538 kg) that were interme¬ diate to Br and Cont Xs. Adj wt for cond did not change rank of breed types but reduced phenotypic diff among breed type slightly. ! LEAFSP0T BACTERIUM TRANSMITTED BY Liriomyza trifolii. Sidney L. Poe, Dept, of .! Entomol . , Virginia Polytechnic Institute and State University, Blacksburg, VA : 24061. Number of bacterial leaf spots (BLS) (Pseudomonas ) on leaves of Chrysan- !| themum stock plants was positively correlated to the number of foliage damage | sites caused by the leafminer, Liriomyza trifolii. Damage sites by caterpillars ! also provided a court through which the bacterium could penetrate plant tissues. Insecticide treatments, fertilizer source and cultivar influenced the number of |l mines and stipples of leafminer and thus BLS. Miner damage was greater on I plants fed NH4 NO3 than on plants fed Ca NO3. Iceberg, Indianapolis White, Improved Rivalry and Nob Hill showed decreasing levels of leafminer attack. An increase by 10 in the number of leafminer and caterpillar damage sites, result- I ed in a corresponding increase of six and seven BLS respectively. Bacterial leaf spot appears to require insect injury for entry into the plant. I | EFFECT OF METHODOLOGIES ON THE COMPOSITION AND PROPERTIES OF DIETARY FIBER I FROM WINGED BEANS. CL Ravindran § J. K. Palmer, Dept, of Food Science, VPI^SU, I Blacksburg, Va. 24061. Winged beans (WB) contain about 25% dietary fiber (DF) , defined as non-starch polysaccharides, and are exceptionally high (Ca 35%) in protein. Dietary fiber has been extracted from winged bean by 5 methods If designed for use on the more typical high-starch, low-protein foods. The DF (' of WB is virtually all "insoluble" fiber, soluble fractions only being obtained j1 when extractions is at elevated temperatures. Galactans predominate in all 1 cases, but the DF also contains substantial glucose (as cellulose), arabinose, xylose, uronic acid, protein, lignin and ash. Considerable differences in the composition and properties of DF fractions resulted from use of the different | methods . 106 THE VIRGINIA JOURNAL OF SCIENCE MAXIMUM YIELD RESEARCH WITH SOYBEANS. Preston H. Reid, D. L. Hallock, Tidewater Research and Continuing Education Center, Suffolk, VA 23437, F. S. Wright*, USDA, ARS, Suffolk, VA 23437 & Norris Powell*, Dept, of Agron., VPI & SU, Blacksburg, VA 24061. Mul tidi sci pi i ne research was conducted to determine maximum returns from economic inputs. Ring Around 604 cultivar yielded 6 bu/A more than Coker 156. Reducing row width from 30 inches to 10 inches increased yields from 46.5 to 61.4 bu/A. Yields increased with plant population to approximately 70,000 plants per acre. No response was obtained by maintaining the soil moisture at 50% plus of the soil available water. In 1981 there was a decrease from irriga¬ tion despite the dry weather. Maximum yields of 68.5 bu/A were obtained with Ring Around 604 cultivar grown in 10 inch rows at population of 102,000 plants per acre. Lodging was severe in all plots and may have limited yields. Soy-, beans planted in killed small grain yielded 64.0 bu/A compared to 58.7 bu/A for those planted in a conventional seed bed. Erosion was less severe in the no¬ till plots. ROW WIDTH, PLANT POPULATION STUDIES WITH SOYBEANS. Preston H. Reid, David A. Morris*, John W. Murphy, Jr.* and John C. Smith, Tidewater Research and Contin¬ uing Education Center, VPI & SU, Suffolk, VA 23437. Soybeans were planted at seeding rates from 33,000 to 300,000 seed per acre and at row widths from 10 inches to 36 inches apart. Significant yield increases attained when row widths were reduced to 20 or 24 inches from 30 to 36 inches. Responses from further reductions in row width beyond 20" were less consistent but did occur on occa¬ sion.' Lodging was increased by both very low and very high plant populations. Yields were not increased by plant population greater than 100,000 but height of first pod bearing node and plant height were increased up to 300,000 plants per acre. Summary of all tests indicates that optimum row width for soybean production is between 10 and 20 inches and optimum plant populations are 100,000 to 150,000 plants per acre. COMMON STALK BORER, PAPAIPEMA NEBRIS (GUENEE) (LEPIDOPTERA : NOCTUIDAE) , CONTROL IN FIELD CORN, ZEA MAYS (LINNAEUS), WITH VARIOUS INSECTICIDES AND A CULTURAL METHOD. James E. Roberts, Sr. and Michael Saluta*, Dept, of Entomol¬ ogy, VPI&SU, Blacksburg, VA 24061. At-planting and rescue treatments and a semi-conventional cultural method were applied in 1982 to no-till field corn in Blacksburg, VA. The test site was arranged in a randomized complete block design, measuring 4 rows (38 in. spacing) X 50 ft. X 4 replicates. The preplanting cultural treatment consisted of mowing the rye cover crop followed by tillage to a depth of 3-4 in. The granular treatments (Lorsban 15G) were applied at-planting. May 17. The fo¬ liar rescue treatments (Ammo® 2.5 EC, Dyfonate® 4E, Imidan® 50W, and Pounce® 3.2 EC) were applied Jun. 4 to the 3 leaf stage corn with a compressed air sprayer delivering 20 gal. /A. Stalk borer damage was evaluated for all treat¬ ments Jun. 13 by using a random visual plant damage rating scale. A moderate to heavy stalk borer infestation prevailed throughout the test area at the time of rescue treatments. Analysis indicated that feeding damage in the untreated control was significantly greater than the treated plots (P< 0.05), however yield differences were found to be nonsignificant (P>0.05). THE VIRGINIA JOURNAL OF SCIENCE 107 CONTROL OF THE HOG LOUSE, HAEMATOPINUS SUIS (LINNAEUS) (ANOPLURA : HAEMATOPINADAE) , ON BROOD SOWS, SUS SCROFA (LINNAEUS), WITH ECTRIN™ 8% EAR TAGS. James E. Roberts, Sr. and Michael Saluta*, Dept, of Entomology, VPI&SU, Blacksburg, VA 24061. Four tests were conducted in Virginia from Dec. 1981 through Feb. 1982 for control of hog lice on brood sows with Ectrin" 8% ear tags. All treated animals received 2 ear tags, one in each ear. Efficacy of treatments was ascertained by recording in weekly intervals the number of lice present on each animal. All animals were either confined individually in farrowing houses or pastured in an uncontrolled environment. A comparative control group was used in each test. In all tests hog lice populations were significantly reduced after 2-3 weeks. Total lice control was achieved by the 4th week in all but one test. Where tests were conducted in farrowing houses, it was noted that the treated sows' pigs were free of lice. All untreated control animals continued to be infested. A total of 4 tags were lost from 3 animals. No other adverse reactions to the ear tags were noted. EVALUATION OF A SYNERGIST WHEN ADDED TO SYNTHETIC PYRETHROIDS FOR CONTROL OF THE HOUSE FLY, MUSCA DOMESTICA (LINNAEUS) (DIPTERA : MUSCIDAE) , IN DAIRY PREMISES. James E. Roberts, Sr. and Michael Saluta* Dept, of Entomology, VPI&SU, Blacksburg, VA 24061. Prentox® PBO 8™ (91.3% Piperonyl Butoxide Technical) was applied as a 5:1 tank mix with one half recommended rates of Ectiban® 5.7% EC and Ectrin™ 10% WDL, Jul. through Sep., 1982, to dairy premises in Montgomery Co., VA. Stand¬ ard rates of both insecticides were also applied to dairies for comparative efficacy. Treatments were applied at separate dairies with a high pressure sprayer (100 psi) . Weekly fly counts were made at all locations, using two methods to evaluate populations. Enumeration of flies lighting on a 'Scudder grill' after 1 min. and an Aeroxon® 'Fly Catchers' sticky tape after 45 min. were recorded at specific sites in a premise. Data were averaged on each day in each treatment. The posttreatment analyses indicated that generally there were no signifi¬ cant differences (P>0.05) among those treatments with PBO 8 and those without. In terms of % reduction, the Ectiban & PBO 8 treatment resulted in the most effective control (sticky tape, >59% and Scudder grill, >83%). FACE FLY, MUSCA AUTUMNAL IS DE GEER, AND HORN FLY, HAEMATOBIA IRRITANS (LINNAEUS) (DIPTERA : MUSCIDAE), CONTROL ON BEEF CATTLE, BOS TAURUS (LINNAEUS). James E. Roberts, Sr. and Michael Saluta*, Dept, of Entomology, VPI&SU, Blacksburg, VA 24061. Several insecticides and application methods were evaluated Jun. through Sep., 1982, on 5 separate herds of beef cattle located in Montgomery Co. and Blacksburg, VA. Treatments consisted of forced usage backrubbers and face- rubbers charged with Ravap® EC and Ectiban® 5.7% EC, Ectrin™ 8% ear tags (2 per animal), and a Dustacator® charged with Ectiban® 0.25% dust. All herds were within ca. 2 miles of each other. Weekly fly counts were made on 10 animals per herd for the duration of the fly season. Comparative efficacy was based on an untreated control herd. The Ectiban EC treatment was the most effective in controlling face flies (>70%). The Ectrin ear tags provided good control of face flies for the first 7 weeks (>63%), but their effectiveness varied for the remaining weeks. The Ravap EC and Ectiban dust treatments resulted in poor control of face flies. All treatments provided excellent control of horn flies. 108 THE VIRGINIA JOURNAL OF SCIENCE TRUE ARMYWORM, PSEUDALETIA UNIPUNCTA (HAWORTH) (LEPIDOPTERA : NOCTUIDAE) , CONTROL IN FIELD CORN, ZEA MAYS (LINNAEUS), WITH POUNCE® 3.2 EC AND AMMO® 2.5 EC. James E. Roberts, Sr. and Michael Saluta*, Dept, of Entomology, VPI&SU, Blacksburg, VA 24061. Pounce® 3.2 EC and Ammo® 2.5 EC were applied as rescue treatments to no-till seedling field corn in Montgomery Co., VA. The test site was arranged in a randomized complete block design, measuring 4 rows (36 in. spacing) X 50 ft. X 4 replicates. The treatments were applied Jun. 15, 1982, with a compressed air sprayer delivering 20 gal. /A. Pretreatment data were recorded Jun. 15. Dam¬ age levels were calculated as the mean percentage of plants per treatment show¬ ing evidence of true armyworm feeding. Post treatment data were recorded Jun. 18, 21, and 24 from the identical plants in each plot and averaged. True armyworm feeding damage was moderate to heavy throughout the test area at the time of treatment. Posttreatment analysis indicated a significantly greater mean damage increase of 23% in the untreated control (P<0.05). The rescue treatments were effective and did not vary significantly in curtailing further armyworm feeding (<5% mean damage increase). FEEDING PREFERENCE OF HAWTHORN LACE BUG. P. B. Schultz. VA Truck & Ornamentals Res. Stn., Va. Beach, VA 23455- Five species/cultivars each of Cotoneaster and Pyracantha were caged in 1981 and 1982 with adults of the hawthorn lace bug, Corythucha cydoniae (Fitch), to evaluate feeding preference. Both terminal and non-terminal cuttings were used in laboratory studies while Cotoneaster were evaluated in outdoor cage studies in 1982. C_. horizontalis and .P. atalantiodes 'Aurea' were least prefered while C. dammeri 'Royal Beauty' and P. koidzumii 'Ingleside Crimson' were the most prefered cultivars. There was no difference between terminal and non-terminal cuttings. C. horizontalis developed lower lace bug numbers than the other species/cultivars in the field study. In a related laboratory study using leaf disks, feeding preference between Cotoneaster, Crataegus, Pyracantha and Sorbus, hawthorn lace bug prefered Sorbus regardless of prior feeding. 1982 TILLAGE FIELD TESTS IN SOYBEANS. Easley S. Smith*, David H. Vaughan, & John V. Perumpral*, Dept, of Agri. Engr., Va. Tech, Blacksburg, VA 24061 & Preston H. Reid, Tidwater Res. & Continuing Education Ctr., Holland Station, Suffolk, VA 23437. Soybean response to various tillage practices was evaluated when double-cropping in a straw mulch following wheat harvest. Under-row ripping to 6, 10, and 14-inch depths and no-till planting treatments were replicated at three locations in different soils. A once-over operation with a modified three- row toolbar mounted ripper-planter was used. Fuel requirements increased con¬ siderably at the deeper tillage depths. Seed germination and stand counts were slightly higher in the deep tilled plots. Harvest yields were similar for all treatments in Turbeville and Woodstown fine sandy loam soils of Goochland and Middlesex Counties, respectively. Increases of 2-7 bu. per acre were noted in deep ripped as compared to no ripped plots in Emporia loamy sand soil of Greens¬ ville County. Disking prior to ripping and planting gave no desirable yield response. (Supported by funds from the Va. Agri. Fndn. and Va. Soybean Commission.) SOUTHERN CORN R00TW0RM CONTROL IN PEANUTS IN SIX SOUTHEAST VIRGINIA COUNTIES. J. C. Smith, Tidewater Res. Ctr., VPI&SU, Suffolk, VA 23437. Research/demonstra¬ tion plots in six southeastern Virginia counties were established in July 1982 for control of the southern corn rootworm, Diabrotica undecimpunctata howardi Barber. Efficacy ratings from best to worst were: Furadan 15G @ 2.0 lb ai/acre, Dyfonate 10G @ 2.0 lb ai/acre, Thimet 15G @ 2.0 lb ai/acre, Furadan 4F @ 2.0 lb ai/acre, Mocap 10G @ 2.5 lb ai/acre, Lorsban 4E @ 2.0 lb ai/acre. Untreated Control. Average % injured pods ranged from 3.4 to 21.2. Yields of peanuts from treatments ranged from 3875 (Untreated Control) to 2879 lb/acre (Furadan 15G). The order of decending yields was: Untreated Control, Lorsban 4E, Furadan 4F, Lorsban 15G, Thimet 20G, Mocap 10G, Dyfonate 10G, Furadan 15G. Values (grade X yield) were in the same sequence as yield. Yield and value parameters did not parallel efficacy results. THE VIRGINIA JOURNAL OF SCIENCE 109 IA MICROELECTROPHORETIC TECHNIQUE FOR DETERMINING SPECIES AND POPULATION AFFIL¬ IATIONS OF SINGLE INSECT EGGS. M.W. Varn, D.G. Pfeiffer, and F.W. Ravlin. Dept, of Entomol. VPISU. Blacksburg, Virginia 24061. An SDS-PAGE (sodium dode- cyl sulfate microslab linear gradient polyacrylamide gel electrophoresis) technique adapted from Matsudiara and Burgess (1978) was used to separate the proteins of single eggs of several species of apple aphids and several popu- [ lations of gypsy moths. This was accomplished by using a .45- .75 mm gel followed by either a coloring (Sammons et al. 1981) or a non- coloring silver : stain (Merril et al. 1982). This technique allows the identification of pest aphid species previously indistinguishable prior to egg hatch; and thus permits pest managers to anticipate control measures. The technique also allows the i mapping of patterns of genetic variability of gypsy moth populations of various densities. BIOLOGY OF MEGACERUS DISCOIDEUS (SAY) ON HEDGE BINDWEED AND ITS SYNCHRONIZATION ! WITH THE HOST PHENOLOGY IN SOUTHWESTERN VIRGINIA. R. Wang* and L. T. Kok, Dept, i of Entomology, Va. Polytechnic Inst, and State Univ. , Blacksburg, Va. 24061. Megacerus discoideus (Say) (Coleoptera: Bruchidae) , a seed feeder, is considered ! one of the most promising biocontrol agents of the hedge bindweed, Convolvulus J sepium L. Studies of this bruchid conducted in southwestern Virginia in 1982 revealed good synchronization between the life cycle of the insect and the phenology of hedge bindweed. M. discoideus has 4 larval instars and one genera¬ tion a year. It overwinters in the fourth instar. The adults emerged from late June to late July. Oviposition peaked during formation of the bindweed seedpods. The larvae burrowed into the young seedpods within 24 hours after eclosion, consuming about 40-50% of the seed material by the time the host plant I becomes senescent in early Sept. Larvae diapaused in September, and fed on the remaining seed material after diapause in spring. Although several larvae were found within a seed, only one could complete its life cycle in each seed. Cannibalism was most common among 2nd instars. HORMONAL REGULATION OF PROGESTERONE SECRETION .BY THE OVINE PLACENTA. S.M. Zirkle, S.J. Philpott and W.E. Beal, Dept, of Animal Science, Va. Polytechnic Inst, and State Univ., Blacksburg, Va. 24061. This experiment was designed to determine the factors controlling placental synthesis of progesterone (P4) . On day (d) 65 of gestation, ewes in Group I were fitted with an implant containing P4 (N=4) or a synthetic P4 (N=8) . Each ewe was ovariectomized (OVX) on d 75 and implants were removed on d 120. Eight ewes were assigned as either intact or OVX controls. Blood samples were collected throughout gestation with frequent sampling on d 65-85 and d 119-125. Ten ewes in Group II were OVX on d 75 and were either injected with saline or 1000 I.U. of human chorionic gonadotrophin (HCG) on d 80 and 115. Group II ewes were intensively bled pre- and post-injec¬ tion of HCG. Concentrations of P4 were determined by radioimmunoassay. Plasma P4 in intact ewes averaged 3 ng/ml on d 65-85 with an increase to 8 ng/ml from d 90-105. Between 105-115 the P4 levels in intact ewes increased to 15 ng/ml. Plasma P4 in OVX ewes implanted with P4 decreased to <2 ng/ml post-OVX, however, the subsequent temporal profile of P4 in the plasma was similar to that of I control ewes. Ewes injected with HCG did not exhibit an increase in the plasma concentration of P4 over that of control ewes. Neither negative feedback nor s gonadotrophin administration affected ovine placental P4 secretion. 1 10 THE VIRGINIA JOURNAL OF SCIENCE Astronomy, Mathematics, and Physics Section THE DESIGN, CONSTRUCTION, AND DIAGNOSTICS OF A Nz PUMPED OSCILLATOR- AMPLIFIER DYE LASER FOR VISIBLE AND U.V. SPECTROSCOPY. Aron Bacs * and G.E . Copeland, Dept, of Physics, Old Dominion University, Norfolk, Va. 23508. Details for the construction of a grazing incidence pulsed oscillator-amplifier dye laser are presented. It is constructed on an optical breadboard for simplicity. The major components of oscillator and amplifier are kept to a minimum, and thus easy to align. A 337.1 nm, 300 KW beam from a Nz laser is beamsplit to pump directly the oscillator and time delayed to pump the amplifier. The dye laser produces a pulsed output ( ~ 12 nsec) from 570 to 600 nm with a line width of 0.002 nm and power up to 50 KW using R6G in ethanol. A novel scheme for reducing amplifier superradiant feedback into the oscillator is described, liming is accomplished via a Z-80 microcomputer system controlling a stepping motor that turns the tuning mirror. The dye laser system has been used to in¬ vestigate the opto-galvanic spectrum of neon which can be used for calibration. (Partially supported by NASA grants NAG-1-1 and NCC1-32) MAGNETOSPHERIC ION EROSION OF THE ICY SATELLITES OF SATURN. Lynn A. Barton, Dept, of Nuclear Engineering and Engineering Physics, Univ. of VA. , Thornton Hall, Charlottesville, VA. 22901. The five inner satellites of Saturn are known to be covered with water ice. Data from Pioneer and Voyager indicate that these icy surfaces are exposed to bombardment by energetic ions. In addition, the presence of heavy ions in the vicinity of these satellites implies the existence of a cloud of heavy neutral molecules, presumably water. In this paper I show that sputtering of ice by the plasma ions is a source of heavy ions in the magnetosphere. A LASER/OPTICAL FIBER ROTOR TIMING SYSTEM, Bruce E. Bernard,* Dept, of Physics, University of Virginia, Charlottesville, VA 22901. A precision timing system which utilizes an optical fiber to guide laser light to a reflecting surface on a spinning rotor and trigger an optical sensor which sends a signal to a computerized data acquisition system is described. The timing system is used to measure the period of rotation of a magnetically suspended rotor spinning at ~ 1 Hz which is part of a gravitational experiment. The effects of noise and non- rotational motion of the rotor which limit the resolution of the period measurements are discussed. (This research is supported by NSF Grant PHY80-07948 and NBS Grant G8-9025). INTRODUCING LOG-LOG PLOTS VIA EXPERIMENT. D. Rae Carpenter , Jr. , Dept, of Physics, Va. Military Inst., Lexington, VA 24450. Data taken on AC voltage vs current in incandescent lamps of power ratings from 15 to 200 watts will produce a straight line when plotted as l n V vs Tn I. This shows the mathematical relationship to be V = V0Im. Values of V0 depend markedly upon the power rating. The slope of the plots, m, is reasonably constant over a wide range of power ratings. There are some variations among bulbs produced by different manufacturers. The experiment involves simple apparatus and the data can be taken quickly enough to warrant measuring several bulbs so as to produce a family of curves. Discussion of the meaning of "inter¬ cept" on a log-log plot may be necessary, i.e., the value of V when 1=1. THE VIRGINIA JOURNAL OF SCIENCE 1 SOFT X-RAY EMISSION STUDIES OF SILICON MATERIALS. R. D. Carson* and S.E. Schnatterly, Department of Physics, University of Virginia, Charlottesville, VA | 22901. The silicon L23 x-ray emission spectrum has been used to study the I valence band density of states for several silicon materials. The amorphous silicon spectrum shows broadened features but with a steeper high energy tail compared to that of crystalline silicon. Samples of amorphous silicon, heavily doped crystalline silicon and silicon nitride show a feature near the top of the valence band which is believed to be from conduction band states, j This feature is particularly prominent in silicon nitride which has a 4 volt band gap and emission from states which appear to be in the band gap is evident. The spectra also show dependence on the energy of the electron beam used to excite the x-rays. These results will be presented and discussed. MOVING MATERIAL INTO SPACE WITHOUT ROCKETS. Raymond S. Cheng. * and James S. Trefil,* Dept, of Physics, Univ. of Va., Charlottesville, Va, 22901. Early steps in the humanization of space include transporting large amounts of metal and other building materials to orbital altitudes. As in other applications, conventional rockets would be burdened with carrying their fuel as they ascend. This inherent inefficiency is avoided through the use of electromagnetic launch¬ ers. Fired from these ground-based devices, the payloads drift to desired heights where they are intercepted. Energy profiles and trajectories of i launched bodies are computed using successively more complex models of the earth i and atmosphere. The results indicate that energy savings are significant when I launchers are used. NUCLEAR WAR - IS IT SURVIVABLE? Robert Ehrlich, Dept, of Physics, George Mason Univ., Fairfax, VA 22030. Any answer to this question clearly depends greatly on the definition of "survivable" . However, within very wide limits imposed by the many uncertainties, certain projections based on scientific facts can be made. It is important to try to be objective on this question, since there are often tendencies to either overstate or understate the probable consequences of a nuclear war to support a particular political position. The material presented in this talk is part of an interdisciplinary course on nuclear war which I have been offering at GMU for the past two years. AIR TABLE EXPERIMENTS WITHOUT AN AIR TABLE. Robert Ehrlich, Dept, of Physics, ; George Mason Univ., Fairfax, VA 22030. A new method has been developed for conducting a wide range of mechanics experiments suitable for introductory physics courses. The method utilizes a new device which can be used for many I of the same kinds of experiments that can now be done on an air table. The simple construction of the device will be described, and its advantages and disadvantages compared to the air table will be discussed. Results for ten different experiments including: uniformly accelerated motion, collisions, l Newton's Second Law, the pendulum, simple harmonic motion, conservation of angular motion, and Lissajous figures will be displayed and discussed. 112 THE VIRGINIA JOURNAL OF SCIENCE SOME QUANTITATIVE ASPECTS OF EMISSION COMPUTED TOMOGRAPHY OF RADIOISOTOPES. Zia R. Hashmi*, 9256 Ashland Woods Lane, Lorton, Va . 22079. Computerized tomography has become an important tool in medical radiography for displaying the internal structural patterns of a complex object. Of particular interest is the tech¬ nique of single photon emission computed tomographyl because of its value in nuclear medicine wherein viable physiologic radiopharmaceutical tracers are used for quantifiable radioactive distributions and their emissions. An import¬ ant parameter involved in such techniques is the depth resolution. The concept of specificity was introduced earlier2 for quantification of depth resolution of tomographic devices. The application os such quantitative concepts to single photon emission computed tomography is discussed in this paper. 1. "Emission Computed Tomography: The Single Photon Approach", Bureau of Radio¬ logical Health, HHS Publ. FDA81-8177 (1981). 2. G. Muehllehner & Z. Hashmi, Phys. Med. Biol. 17, 251 (1972). ON THE COULOMB SCATTERING PROBLEM IN THE PRESENCE OF AN INTENSE LASER FIELD. J.C. Huang*, Phys. Dept., Univ. of Missouri, Columbia, MO 65211, and W. Majewski*, Phys. Dept., Univ. of Maryland, College Park, MD 20742. A comparison to the lowest order in an intensity dependent dimensionless coupling constant is made between the semiclassical Volkov and QED approaches to the Coulomb scattering problem in the presence of a high intensity external E & M field. In the nonrelativistic infrared limit, the elastic, one photon emission and absorption cross sections are essentially the same. However, the relativistic comparison indicates differences which are more pronounced. EXCITATION SPECTRUM OF A ONE-DIMENSIONAL ANTIFERROMAGNET AT FINITE MAGNETIZA¬ TION. Michael D. Johnsonf Dept, of Physics, U.Va. , Charlottesville, VA 22901. In developng a quantum theory of ferro- and antiferromagnetism, the intrac¬ tability of the three-dimensional theory spurred the study of simplified one¬ dimensional models. More recently, these idealized systems have become in¬ teresting in their own right, as a result of two developments: the manufacture of quasi-one-dimensional magnetic materials, and the discovery that the one¬ dimensional magnetic model lies in a larger class of exactly soluble one¬ dimensional many-body systems. The state of a system is described quantum mechanically as a set of allowed energy levels; at low temperatures, only the lowest energy states are of physical interest. We have computed these elemen¬ tary excitations (at zero temperature) for an anisotropic (xxz) one-dimensional antif erromagnet in the presence of an external magnetic field. EPR SATURATION RECOVERY IN MYOGLOBIN. Paul D. Levin* and A.S. Brill*, Biophysics Program and Dept, of Physics, Univ. of Va. , Charlottesville, Va. 22901. Between 2K and 4K the recovery from power saturation of the EPR signal of myoglobin is mostly due to the two phonon Orbach process with some contribu¬ tion from the single phonon direct process. The recovery curve is not a single exponential function but arises from a distribution of single exponential re¬ covery signals based on a distribution of characteristic relaxation times, T-^. The experimental recovery curve may be fitted to a single exponential function which provides an effective relaxation time, T]_, the temperature dependence of which can be analyzed in terms of a four-level model of the heme iron based on distributions in state energies. Experimental and simulated data have been compared by means of a Marquardt non-linear least squares fitting routine, and, currently, a linear polynomial expansion in temperature. The energy fluctua¬ tions obtained in this way are in agreement with those determined from the analysis of independent measurements. THE VIRGINIA JOURNAL OF SCIENCE 1 ON A NEW PARITY-VIOLATING EFFECT IN ATOMS. Walerian Majewski*, Phys . Dept., Univ. of Maryland, College Park, MD 20742. We show that the electric part of the nuclear current produces a short-range magnetic field inside the nucleus. This field is usually either neglected, or is considered to be vanishing automatically. It can be parametrized in terms of nuclear static transverse electric multi pole moments E, . The moments with even 1 are forbidden by T-invariance, and those with Aad 1 are forbidden by P-invariance. The influence of the nuclear parity impurity on the parity impurity of atomic levels is estimated. SUPERCONDUCTIVITY OF EXTREMELY SMALL TUNGSTEN CARBIDE PARTICLES. P.M. Miller, M.T. Lough and B.S. Deaver, Jr., Dept, of Physics, Univ. of Va. , Charlottesville VA, 22901. The superconductivity of cubic WC particles of mean diameters 10, 6 and 3 nm is being studied by measurements of the magnetization as a function of temperature and magnetic field to search for possible size effects. Measure¬ ments are being made with a SQUID susceptometer and with an ac technique on powder samples with connectivity among the particles and on particles dispersed in an insulating matrix. Fits to theoretical curves provide values for the tran¬ sition temperature Tc, critical field and gap parameter. For all the samples Tc - 6 . 5 K. A FORMULATION OF RIGID-BODY ROTATIONAL DYNAMICS BASED ON EULER PARAMETERS AND CONJUGATE GENERALIZED ANGULAR MOMENTA. Harold S. Morton Jr. t Dept, of Mech. and Aero. Engineering, University of Virginia, Charlottesville, Va. 22901. The orientation of an arbi¬ trary rigid body is specified in terms of a set of four Euler parameters. A correspond ing set of generalized angular momentum variables is derived and then used to replace the usual angular velocity vector in specifying the time-rate-of-change of orienta¬ tion relative to inertial space. This leads naturally to a formu¬ lation of rigid-body rotational dynamics in terms of a system of eight first-order coupled differential equations involving the four Euler parameters and the four conjugate angular momenta. The equations are expressed in matrix form and exhibit a remark¬ able symmetry, which represents the key result. Euler's equations are used in the derivation, and the results are, of course, con¬ sistent with Hamilton's equations. THE PHYSICS OF A RELATIVISTIC OSCILLATOR. W. Barlow Newbolt, Dept, of Physics, Washington and Lee University, Lexington, Va., 24450. Because of its usefulness in teaching the special theory of relativity, the properties of a relativistic oscillator are described in this paper. Two features of the oscillator are emphasized: its mechanics and its usefulness as a device to measure time intervals. INTERACTION OF NEPTUNIUM-239 CD WITH DOG SERUM IN VITRO. David A. Petersen, i Dept, of Chemistry and Geology, Mary Washington Col., Fredericksburg, Va. 22401, Dr. R. A, Guilmette and Dr, M. A. Medinsky, Inhalation Toxicology Research In¬ stitute (ITRI) , Albuquerque, N. M. Serum Protein Binding or association of I Np-239 (pentavalent state) was clearly indicated by gamma and U.V. spectroscopy I of various Np incubated serum protein samples. Protein bound Neptunium Frac¬ tions were separated by TCA precipitation, dialysis, ultrafiltration, and gel chromatography methods. Ratios of activity present in the protein bound frac¬ tions to non-bound fractions were used for this determination. (This work was completed at the ITRI in Albuquerque, N. M. under an A.W.U. grant.) 114 THE VIRGINIA JOURNAL OF SCIENCE A SENSITIVE TEST OF THE SECOND LAW OF THERMODYNAMICS. C. B. Satterthwaite, Dept, of Physics, Va. Commonwealth Univ. , Rich¬ mond, Va. 23284. The accompanying sketch is a prototype of a familiar perpetual motion machine of the second kind. The column of water in the reentrant tube, which is filled with tiny glass beads, is held up by surface tension. The water surfaces in the pores of the sintered glass disc at the top are, therefore, concave and have a lower vapor pressure than a flat surface. Naively, one might think that water would evaporate from the flat surface and condense in the pores causing a con¬ tinuous flow of water. This complete conversion of heat energy to mechanical energy, of course, violates the second law of thermodynamics and can be shown not to work if one considers the gradient of vapor density in a gravitational field. It is, nonetheless, a sensitive test of the second law. Li nH AND LiisH CLUSTER MODELS OF MUON PROBES IN BCC METALS. D. D. Shill ady and P. Jena, Depts. of Chem. and Physics, Virginia Commonwealth Univ., Richmond, Va. 23284. Hartree-Fock-Roothaan self-consistent-field calculations have been car¬ ried out for LinH and Li 1 5 H using a (3G1S/1G2S/1G2P) basis for Li and (4G1S/ 2G2S/1G2P) for H as lobe-mimics of spherical harmonics. The energy difference between Oh and the favored Td sites for H depend on the size of the cluster studied; in Lii5H this difference was found to be 0.136 ev. The barrier to diffusion along the (1,1,1) direction is prohibitive relative to Oh-Td hopping. In a vacancy model (Lii5H) H moves away from the vacancy and Td sites are still lowest in energy, but the Oh-Td barrier is less, allowing enhanced mobility around the vacancy. In the chosen basis a symmetric Li 14 vacancy cluster con¬ tracted 11.4% in spite of the fact thatcthe same basis predicts an 8.0% larger lattice constant than experiment (3.772A calc. vs. 3 . 491 A exp.). These results predict a local lattice contraction about a vacancy with increased H mobility around the relatively repulsive vacancy site. CHARACTERIZATION OF CaF? FILMS DEPOSITED ON SINGLE CRYSTAL GE SURFACES. BILLY W. SLOOPE, Dept, of Physics, Va . Commonwealth Univ., Richmond, Va. 23284. CaF« films were deposited by vacuum evaporation onto (100), (110), (Ill), and (112) Ge surfaces at various deposition temperatures and rates and to various thick¬ nesses. Structure of the films was predominantly polycrystalline with an increasing (111) texture at deposition temperatures above 330°C. Continuity, grain size, and index of refraction also showed dependence on deposition temperature. Current-voltage measurements were made by depositing a small Ag electrode on top the CaF2 film. The I-V characteristics were generally non-symmetrical with respect to current direction, mostly non-linear, yielding no single power relationship, and at higher currents exhibited time variations in voltage. These characterizations will be summarized and some potential future directions of research discussed. THE VIRGINIA JOURNAL OF SCIENCE 115 A RADIATIVE TRANSFER MODEL FOR REMOTE SENSING OF LASER INDUCED FLUORESCENCE IN STRATIFIED MEDIA. D. D. Venable, A. R. Punjabi*, Dept, of Physics and 1 Engineering Studies, Hampton Institute, Hampton, Va. 23668. A semi-analytic Monte Carlo radiative transfer model (SALMON) for oceanographic lidar systems |j has been applied to inhomogeneous media. The values of the ratio of the fluorescent wavelength signal (H ) to the Raman wavelength signal (HR) are I determined for ambient chlorophyll concentrations ranging from 0.01 ug/f to 20 yg/A. The concentrations vary linearly as a function of depth in the medium with gradients of ± 1% per meter to ± 20% per meter. Comparison of the j; ratio Hp/H for homogeneous (zero gradient) and inhomogeneous cases show significantly large deviations at low concentrations and high gradients. The ji deviations decrease as concentrations increase or as gradients decrease. In I our model, the surface of the medium is flat and no lateral inhomogeneities are allowed. (Supported by the National Aeronautics and Space Administration) X-RAY TOPOGRAPHY OF NEARLY PERFECT CRYSTALS USING A MONOLITHIC DIFFRACTION SOURCE. R. F. Zi 1 haver* and W. C. Sauder, Dept, of Physics, Va. Military Inst., Lexington, VA 24450. A new method of x-ray reflection topography is proposed. The Berg-Barrett method, double crystal technique of Bonse and Kappler, and the mismatched crystal arrangement of Deslattes, Torgeson, Paretzkin and Horton are all compared with the ideal topographic experiment. It is shown that the use of a monolithic double crystal spectrometer i (MDCS) to condition the x-ray beam could provide a closer approach to the ideal. The new technique will permit the examination of as much as one square centimeter of the surface of a high per- f ecti on crys tal . Biology Section A COMPARATIVE STUDY OF THE EFFECTS OF TEMPERATURE ON RATES OF DEVELOPMENT IN ! SEA URCHINS. W. Bruce Adams* and Carolyn M. Conway, Dept, of Biol., Va. Commonwealth Univ., Richmond, Va. 23284. The rate of embryogenesi s in sea i urchins is temperature dependent. We examined the influence of temperature on development of Arbacia punctulata. Rates of development from fertilization I through the pluteus larva stage were monitored at three temperatures (20°, 25°, 1 and 30° C) in order to determine the exact relationship between these tempera¬ tures and rates of embryogenesi s. Our studies indicated that at 25° C develop¬ ment proceeded faster than at either 20° C or 30° C. The rate of development 1 at 30° C was somewhat faster than the rate of development at 20° C. The j occurrence of a maximum rate of development at 25° C indicated that an optimum i temperature for Arbacia punctulata embryogenesi s may occur within the tempera- ( ture range studied. THE EFFECTS OF DEXATRIM DIET CAPSULES ON MICE. D.D.Alvis*, A.F. Conway, and P.L. Dementi, Dept, of Biol., Randolph-Macon Col., Ashland VA 23005. Forty male white mice were grouped by age and then paired by weight. Five mice in each age group received 1.5 mg of phenylpropanolamine hydrochloride per kg of body weight in their water bottles; five received 3.0 mg/kg of body weight; and ten were controls. Body weight and water consumption were monitored daily for one month. At the end of this time period, the animals were sacrificed and their hearts, livers, and kidneys were removed and weighed. All animals gained weight during the experiment. A paired t-test revealed no significant differences in body weight gain or organ weights between experimental and control animals. Water intake by experimental mice was significantly greater than in controls. 116 THE VIRGINIA JOURNAL OF SCIENCE THE ROLE OF THE ADRENAL CORTEX IN FAT METABOLISM OF THE MONGOLIAN GERBIL, (Meriones unguiculatus) . Safia Baggia* and Dr. Frank Leftwich, Dept, of Biology, Univ. of Richmond, Richmond, Va. , 23173. Sixty four male Mongolian gerbils, Meriones unguiculatus were adrenalectomized and treated with daily injections of cortisol (.lmg) and aldosterone (50yg) . Upon necropsy, depot fat was found to be significantly lower in the hormone treated groups. Blood free fatty acid was found to be significantly higher in the controls than in treated gerbils. Significant differences were also found in survival time and in the daily percent change in body weight of hormone treated groups. PHENOBARBITAL TREATMENT DURING THE EARLY NEONATAL PERIOD RESULTS IN INCREASED HEPATIC P-450-DEPENDENT MONOOXYGENASE ACTIVITY IN ADULT MALE AND FEMALE SPRAGUE- DAWLEY RATS. Daniel M. Bagley* and Johnnie R. Hayes*, Dept, of Pharmacology and Toxicology, Medical College of Virginia, Richmond, VA 23298. Phefiobarbital (PB) was injected subcutaneously to neonatal male and female Sprague-Dawley rats once on each of the first five days after birth to determine the long term consequences of this treatment on the cytochrome P-450-dependent monooxygenase system. At 20 weeks of age there were no differences between treat ment and control groups of either sex in liver weight as a percent of body weight and in mg. protein/g liver. However, both male and female PB treated groups showed significantly higher P-450 content, P-450 reductase activity, ethoxycou- marin-O-deethylase activity and 4-methylumbilliferone glucuronidation relative to ‘controls . These findings suggest that xenobiotics which typically have only transi¬ tory inductive effects in mature animals may cause permanent effects if the exposure occurs during early neonatal life. ANDROLOGICAL EVALUATION OF MEN INFECTED WITH UREAPLASMA UREALYTICUM. William Ballard and Steve Ackerman, Andrology Laboratory, Dept, of Biol., Old Dominion Univ. , Norfolk, Va. 23508. Several recent investigations have implicated reproductive tract infections of Ureaplasma urealyticum with abnormal semen parameters. In this study, semen samples from 134 males undergoing fertility evaluation from June 1 to October 31, 1982 were examined to determine if alterations in specific semen characteristics were associated with the presence or absence of IJ. urealyticum. Fifty-four (40.3%) of the specimens cultured positive for U. urealyticum. No significant differences were observed between infected and uninfected specimens for semen volume, sperm count, motility or morphology using the Student's t-test or nonparametric statistical analyses. These data confirm a previous investigation by our laboratory (Fertility and Sterility 36:648, 1981), but contradict two earlier reports implicating an adverse effect of U. urealyticum infection upon sperm motility. Investigations on the effects of the titre of U. urealyticum infection on semen parameters are presently being undertaken. EFFECTS OF FEEDING AND DIAPAUSE ON OVARY DEVELOPMENT IN TIGER BEETLES. Betty E. Berry & C. Barry Knisley. Dept, of Biology, Randolph-Macon College, Ashland, Ba . 23005. A laboratory study of the effects of prediapause and postdiapause feeding on winter survival and ovary development was conducted with Cicindela tran- quebarica and Cincindela repanda . Beetles fed at low levels prior to diapause had a significantly higher mortality during diapause than beetles fed ad libidum. A period of diapause was found necessary for females to reach sexual maturity. Feeding during postdiapause must remain at high levels to insure proper ovary development. Beetles fed at low levels before dia¬ pause gained a higher percentage of weight in postdiapause than beetles at other feeding levels, and they also showed ovary maturation and egg production equal to beetles fed ad libidum during prediapause. Thus low feeding levels in the fall can apparently be compensated for by high feeding in the spring. THE VIRGINIA JOURNAL OF SCIENCE 117 FECAL COL IF ORM AND FECAL STREPTOCOCCI ENUMERATION IN THE GREAT DISMAL SWAMP. Nancy W. Bourgeois* , Dept of Biol., Old Dominion University, Norfolk, Va . 23508. Fecal coliform and fecal streptococci enumeration was done in the Great Dismal Swamp in Lake Drummond and Feeder Ditch during the months of June, July, and August. It was found that Lake Drummond is free from fecal coliforms. Fecal coliforms and fecal streptococci in Feeder Ditch increased from Lake Drum¬ mond to the Inland Waterway, suggesting that their source is land run-off through drainage pipes which were sampled. Increased rainfall corresponded to increased bacterial counts, supporting this idea. Fecal coliform to fecal streptococci ratios suggest that the land run-off is primarily from non-human storm-water run-off. Counts obtained for this study may be lower than counts obtained in a spring study due to higher water temperature and decreased rainfall and current flow in the summer. A STUDY OF VARIATION IN EASTERN TIMBER RATTLESNAKES, CROTALUS HORRIDUS. Christopher W. Brown*, Dept, of Biol., Geroge Mason Univ., Fairfax, Va. 22124. Variation was examined in specimens of Crotalus horridus from the eastern United States in an attempt to substantiate the status of its two described subspecies as defined by Gloyd (1940). A particular effort was made to dupli¬ cate the results of a study by Pisani, et al. (1973), who concluded that no valid subspecies exist. A maximum likelihood factor analysis and stepwise discriminant analysis were performed on the same morphological characters used by Pisani, et al. (1973). Several additional characters relating to size and pattern were also examined. DISPERSAL AND DISPERSION IN A POPULATION OF SOLDIER BEETLES, CHAULIOGNATHUS PENNSYLVANICUS . James R. Brown* and Luther Brown, Dept, of Biol., George Mason Univ., Fairfax, Va . 22030. Mark-recapture methods were used to examine (movement patterns in a population of soldier beetles (Chau! iognathus pennsylvanicus) . Eighty-one percent of 776 marked beetles were recaptured 24 hours after release. Frequency distributions for distances moved were simi¬ lar and leptokurtic for both sexes. Abundance of both males and females with¬ in the study site was highly correlated with presence of breeding sites and food (Sol idago sp. ) . Partial correlation analyses suggest that males and females responded to Sol idago plants in different ways: male density was posi¬ tively correlated with plant abundance, female density was positively cor¬ related with male abundance, and uncorrelated with plant abundance. Pro¬ portions of each sex remaining at a site were correlated with the density of the opposite sex, but uncorrelated with Sol idago . VARIABILITY IN DAPHNIA ACUTE TOXICITY DATA. Arthur L. Buikema, Jr., University Center for Environmental Studies and Department of Biology, Va . Polytechnic Institute and State University, Blacksburg, Va. 24061. Three independent labs acutely exposed Daphnia magna to 5 industrial effluents using a draft test protocol. Comparisons were made of percent survivors after 24 hr exposure to 100% effluent and 48-hr toxicity data. There was considerable intra- and inter¬ lab after 24 hr. Little intralab variability in 48-hr data was noted; however, there were significant interlab differences in 48-hr LC50s and dose-response slopes. Variation in results may be due to differences in test and culture conditions and genetic differences among animal stocks. Subcultures of the daphnid stocks were obtained and cultured under identical conditions for 50+ generations. Chromium exposed animals did not exhibit similar dose-response slopes suggesting that genetic differences may be a major factor in contributing to the variability in organism sensitivity to pollutants. 118 THE VIRGINIA JOURNAL OF SCIENCE SIZE -RELATED PATTERNS OF OVARIOLS AND ECG NUMBERS IN A COMMUNITY OF TIGER BEETLES (COLEOPTERA: CICINDELIDAE) . MARK C. CARTER AND C. BARRY KNISLEY. Dept, of Biology, Randolph-Macon College, Ashland, Va. 23005. Body size was strongly correlated with ovariole number and mature egg size in fifteen species of Cicindela collected in southeast Arizona. Ovariole number was strongly correlated with reproductive number. These relationships suggest greater reproductive potential in larger beetles. No relationship between adult population size and reproductive potential was found, but that adult populations were dependent on habitat type. This indicates reproductive cell number is strongly environmentally influ¬ enced while ovariole number is constant within species. THE EFFECTS OF PH ON SURVIVORSHIP AND GROWTH RATE IN THE FINGERNAIL GLAM, MUSGULIUM PARTUMEIUM (SAY) (PIVALVIA: PISIDTIPAE). Daniel L. Childers* & Daniel J. Hornbach, Dept. Piol., Univ. of Va., Charlottesville, Va. 22901. The effects of pH on survivorship, growth and natality of a stream population of M. partumeium were examined in the laboratory by maintaining cultures of clams at pH 3,4,5, and 6 with H2SO4 and a control (no acid; pH = 7). For indivi¬ duals collected in October, the median survival times were 10.5, 24.0, 41.1, 47.1 and 62.1 days for pH 3,4, 5,6, and 7 (control), respectively, indicating that acidification had a great effect on survivorship. Clams maintained at pH 3 or 4 showed no growth (based on measures of shell length), whereas, clams at pH 5 displayed higher growth rates than those at pH 6 or 7 (control) (0.11, 0.07 & 0.06 mm/wk for pH 5,6 & 7, respectively). Pirth rates averaged 11 & 1 hirths/individual for 94 & 64 days for clams maintained at pH 5 and 7, respectively; no births were recorded for other treatments. This indicates that both rates of growth and reproduction are affected hy acidification and that maximum rates are seen at a pH near that of the environment from which these individuals were collected (river approx. pH 5.2). Preliminary data on animals collected in January show the same general trends, but with variations in the specific survivorship, growth, and reproductive rates. LIFE HISTORY TRAITS OF TWO POPULATIONS OF THE FRESHWATER CLAM, PISIDIUM CASERTANUM (POLI) (PIVALVIA: PISIDTIPAE) IN TWO SOUTHWEST VIRGINIA PONDS. J\ Oox* and D.J„ Hornbach. Dept. Piol., Univ. Va., Charlottesville, Va. 22901. The 2 populations (FP & RP) studied inhabit ponds with different H2O chemis¬ try (e.g., conductivity 240 & 15 ymho for EP & RP respectively) and display different life history characteristics. Shell lengths (SL) range from 0.7 to 3.3 & 4.8 mm for RP and EP respectively. Transfers were conducted to test for environmental influences on traits. For clams 1.3^SL<2.0 birth rates were eaual in controls (RP-*RP, EP-*EP) while for clams >2.0 birth rates EP->EP>RP->RP indicating greater overall fecundity in EP. Pirth rates in EP+EP>EP-»RP and RP->EP>RP-»RP. Mortality was greater in EP-*RP than in other transfers for all sizes of clams. These results indicate environment influences life history traits. Clams were cultured in waters of various hardness to test if increased Ca availahilty was responsible for observed life history difference. Mortality was greatest in softer waters, with RP>EP except in very soft where mortality was high in both EP & RP. Growth rates for clams SL<1.2 mm was greater in hard and control treatments. Thus, H2O hardness affects life history traits. Electrophoresis indicates that there could he a genetic component to observed differences. RP has 1 genotype; EP has 2, with one being the RP type. THE VIRGINIA JOURNAL OF SCIENCE 119 EFFECTS OF 6-METH0XYBENZ0XAZ0LIN0NE (6-MBOA) ON SEXUAL MATURATION IN SEVERAL RODENT SPECIES UNDER DIFFERENT PHOTOPERIODI C CONDITIONS. J. A. Cranford , Biol ogy Dept., V.P.I. & S.U., Blacksburg, VA 24061. Photoperiod and the influence of 6-MBOA on sexual maturation was evaluated on several small rodent species (Pero- myscus leujopus, P_. maniculatus, Microtus pennsyl vanicus and M. pi netorum) . All species were tested under long (LD 16:8) and short (LD 8:16) photoperiods at 45 days of age. Under long photoperiod, 3 species were additionally evaluated at 30 and 35 days of age to examine age dependent effects. Minimum number of ani¬ mals per group was 20 controls and experimental s of each sex and each photo- periodic condition. Animals were intraperi toneal ly injected with 0.5cc of con¬ trol of experimental (5 pg 6-MBOA) solution for 3 days with body weight, adrenal glands and reproductive organs assayed on the fourth day. Results showed a positive response for all species on long photoperiod while short photoperiod results varied. Those species tested at different ages indicate a strong age dependency in the response. Food consumption of long photoperiod groups was not different between controls and experimental s nor was body or adrenal weights. Experimental animals had lower body fat levels (Soxhlet extracted) than controls indicating that energy for sexual maturation was derived from existing fat energy reserves. LOW TEMPERATURE EFFECTS ON GROWTH AND BODY WEIGHT IN MICROTUS PINETORUM. J. A. Cranford and N. Thumser, Biology Dept., V.P.I. & S.U., Blacksburg, VA 24061. The effect of cold ambient temperatures on growth, body weight dynamics and food consumption was determined for juvenile and adult Microtus pi netorum. Each age group was monitored through a warm temperature phase (20°C) followed by a short low temperature phase (1-5°C) and then returned to warm temperature. Animals were either housed singularly or multiply by sex to determine if huddling behav¬ ior would reduce the effect of low temperature exposure. Juveniles and adults differed in their response to cold period exposure, with juveniles showing con¬ tinuous growth while adults lost weight. Singly and multiply housed pine voles differed in body weight maintenance but not in a consistent manner. Food con¬ sumption was higher in singly housed pine voles compared to multiply housed and during cold exposure both groups showed significant increases in food consump¬ tion. Singly housed cold exposed voles significantly increased food consumption relative to multiply housed voles. Other microtine species have shown body weight responses to photoperiod length changes similar to those observed in re¬ sponse to these temperature changes. Field data and controlled natural environ¬ ment experiments indicate that body weight dynamics are more responsive to thermal changes than photoperiodic changes. BREEDING BIOLOGY OF THE PURPLE MARTIN, PROGNE SUBIS SUBIS, IN WILLIAMSBURG AND JAMES CITY COUNTY, VIRGINIA. Robert R. Cross, Dept, of Biology, Christopher Newport Col., Newport News, Virginia. 23606. The purple martin is a member of the swallow family and breeds colonially in artificial nesting boxes throughout North America. In Williamsburg and James City County, the breeding success of this species was investigated via a banding project. Results indicate that purple martin populations are limited by strong inter¬ specific and intraspecific competition for nest sites. Juvenile purple martins are often harrassed on their initial flight from the nest of subadult martins. Two hypotheses have been proposed to explain this behavior. Implications of the strategy and motivation behind "juvenile harassment" are discussed. 120 THE VIRGINIA JOURNAL OF SCIENCE EFFECTS OF HIGH AND LOW DIETARY PROTEIN LEVELS ON THE DIAMETER OF MUSCLE FIBERS OF THE MOUSE BICEPS BRACHII. L.M.DiClemente*, A. F. Conway, and P.L. Dementi, Dept. of Biol., Randolph-Macon Col., Ashland VA 23005. The level of dietary protein received by mice was varied and the effect on muscle fiber size within the biceps brachii was measured. Eighteen mice were grouped by weight and fed control, high protein, or low protein diets. One group was weighed and sacrificed each day for 6 days. The muscle fibers of the biceps brachii were measured histologically. The mice on control and high protein diets showed moderate fluctuations in body weight while mice on low protein diets progressively lost weight. The mean fiber diameter in mice fed control or high protein diets was 19 microns but was reduced to 16 microns in mice on low protein diets. SUBLETHAL EFFECTS OF THE WATER SOLUBLE FRACTION OF JP-4 JET FUEL ON THE BLUEGILL (LEPOMIS MACROCHIRUS) . Thomas R. Doane *, A. L. Buikema, Jr. & J. Cairns, Jr., Univ. Ctr. for Environ. Stud., Va. Polytechnic Inst. & State Univ. , Blacksburg, Va. 24061. Equipment was developed to continuously deliver a water soluble fraction (WSF) of JP-4 jet fuel for exposure of bluegills. Analytical proce¬ dures were developed to determine the concentration of the WSF. The 96hr-LC50 of the WSF, tested in a flow through bioassay dilutor system, was determined to be 26% of the saturated WSF. A concentration of the WSF which caused a statistically significant shift in the ventilation rate, was 17% of the saturated WSF. Normal levels of ions and enzymes in the blood of the bluegills were determined. Changes in blood ion and enzyme concentrations were noted when fish were exposed to sublethal levels of WSF. Transmission electron micrographs were also taken of the gill and liver tissue from exposed fish. (Supported by a grant from the Air Force Office of Scientific Research.) PARASITES OF THE OPOSSUM, DIDELPHIS VIRGINIANA, IN NORTHERN VIRGINIA. Ralph P. Eckerlin, Div. of Natural Sciences, Northern Va. Comnty. Col., Annandale, Va. 22003 Eleven species of metazoan parasites were recovered from 25 opossums collected in Fairfax Co. in northern Virginia. These included: Trematoda- Brachylaima virginianum; Cestoda- Mesocestoides corti; Nematoda- Capillaria sp., Cruzia americana, Longistriata didelphis , Physaloptera turgida; Acanthocephala- two unidentified immature specimens; Siphonaptera- Ctenocephalides felis, Orchopeas howardii; Acarina- Dermacentor variabilis , and unidentified immature mites. The immature acanthocephalans and mites may be accidental parasites of the opossum. The adult parasite species found are common parasites of the opossum in North America; however, several are new records for Virginia. EVALUATION OF KODAK TECHNICAL PAN AND ILFORD XP-1 400 FILMS FOR ROUTINE BIOLOGICAL PHOTOGRAPHY. J.M. Emmett III* and A. F. Conway, Dept. of Biol., Randolph-Macon Col., Ashland, VA 23005. Two new black and white negative films were evaluated under conditions routinely encountered in biological photography. Kodak Technical Pan was developed in diluted Edwal FG-7 or Kodak Technidol. Ilford XP-1 was developed in the Ilford XP-1 kit. These films were compared with Kodak Panatomic-X and Kodak Tri-X Pan developed in Kodak D-76. Technical Pan developed in Edwal FG-7 or Technidol produced a wider range of gray tones and much finer grain than Panatomic-X. Varying development time in Edwal FG-7 allowed adjustment of Technical Pan contrast to match the tonal range of the subject. Ilford XP-1 displayed more grain and less resolution than Technical Pan or Panatomic-X, but less grain than Tri-X. Resolution and sharpness were equal in XP-1 and Tri-X. THE VIRGINIA JOURNAL OF SCIENCE 121 FINE STRUCTURE OF AN OSMOREGULATORY ORGAN FOUND IN THE MITE FAMILY ALGOPHAGIDAE . Norman J. Fashing , Dept, of Biol., Col. of William & Mary, , Williamsburg, Va. 23185. Mites of the astigmatid family Algophagidae are characterized by an elevated sclerotized band of cuticle on each side of the propodosoma between legs I and II ("axillary organs"). Scanning and transmission electron microscopy of 'these organs in Algophagus pennsylvanicus , a species found in water-filled treeholes, reveals a porous plate covering specialized epidermal cells which contain plasma membrane plications in close contact with numerous mitochondria. Such cells are typically involved in active transport. Morphologically similar structures have been found to function in osmoregulation in aquatic insects, and the same is probably true for the axillary organs of this family of aquatic mites. EFFECTS OF INTRAFOLLICULAR INJECTION OF A VOLUME EXPANDER ON OVULATION IN THE RABBIT. Kathryn P. Flagge and R. J. Swanson, Dept, of Biological Sciences, Old Dominion Univ . , Norfolk, VA 23508. Intraf ollicular injection of solutions into rabbit ovaries V'LVO was accomplished with sharpened, beveled, and smoothed glass microneedles having bore diameters ranging from 5-25 micrometers. Needles were fitted with plastic tubing, which was attached to a 5 microliter Hamilton syringe loaded with the appropriate solution for injection. Surgery was performed on mature female New Zealand rabbits two to three hours post coitus. With an ovary exposed, randomly selected Graafian follicles were either: (1) injected with physiological saline, (2) injected with 2% Blue Dextran (MW 2,000,000) in physiological saline, or (3) left uninjected. All follicles visible at the time of ovarian exposure and any administrations to these follicles were indicated on a reference map used for orientation upon histological examination of the ovary following its removal 18 1/2 to 23 hours postovulation. Evaluation of follicular status (ovulated or non-ovulated) at the time of ovarian excision was performed within and among the test and control groups. Intraf ollicular pressure was not recorded. AN OSTEOLOGICAL OVERVIEW OF THE SEVEN PERCINA SPECIES IN THE SUBGENUS ALVORDIUS (PISCES : PERCIDAE) . Ann S . Fulcher* and W. S. Woolcott, Dept, of Biology, Univ. of Richmond, VA. , 23173. An osteological investigation was conducted to deter¬ mine the phylogenetic relationships among seven Percina species, subgenus Alvordius. Specimens were examined using X rays and an enzymatic clearing and staining technique. Diagnostic skeletal characters included the coracoid, premaxilla, dentary, and hypurals. Preliminary findings indicated that (1) IT maculata is representative of an earlier ancestral stock and (2) IT macrocephala and IT pantherina, which occur west of the Appalachian Mountains, are similar and differ from the eastern species IT crassa, IT no to gramma, IT peltata, and ?_, roanoka which appear morphologically closer to IT maculata. (Supported in part by a University of Richmond Undergraduate Grant) 122 THE VIRGINIA JOURNAL OF SCIENCE DIURNAL AND SEASONAL INFLUENCES ON THE METAPOLIC RATE OF THE ASIATIC FRESHWATER CLAM, CORPICULA FLUMINEA (MULLER) (BIVALVI A: COPPICULIPAE) . S.E. Gardner* & D.J. Hornbach, Dept, of Piol. , Univ. of Va., Charlottesville, Va . 72901. Metabolic rates (Vp2) were measured on a daily basis at 3 periods throughout the sprinq (groups if II, & III collected 31 March, 22 April, 10 May 1982, respectively;. Significant van' aMnnc in Vr, n^nr <-,n a Haiiw hocig (as much as 76 % for 2 mg periods of high and 2 clams of group periods of low activity at approximately 1300 & 2000 h and 0900 & 1700 h, respectively. Daily changes in Vp2 may he associated with energy reouiring osmoregulation since variations and rhythmic; ty of reported rates coincide. It is also possible that the diel patterns reported are representative of a portion of the circadian rhymicity in physiological functions of Corhicula. Greatest Vo2 and the most pronounced diel rhythms were seen in group II, while lowest Vpu (as much as 79% below rates for group II clams) and smallest variations were recorded for group III. Shell length-ash free dry weight relations changed during this study with group III showing more tissue weight per unit shell length than group I. It is suggested that this might he the result of this population becoming reproductively active. The changes in Vq2 from group to group are probably the result of the clam's changing reproductive status. THE USE OF OTOLITHS FOR DETERMINING DAILY AGES OF YOUNG-OF-THE-YEAR SMALLMOUTH BASS. Robert J. Graham*, Dept, of Fish, and Wild!. Sci., Va. Polytech. Inst, and State Univ., Blacksburg, Va. 24061. Viable smallmouth bass eggs were collected from a wild nest on 26 May, 1982 and hatched under laboratory con¬ ditions to determine if the growth rings on the saggitae of young-of-the-year smallmouth bass were daily rings as suggested by previous otolith research. Ten smallmouth bass were removed from the laboratory culture every other day from the time of swim-up until 14 days after swim-up, frozen, and the saggitae later examined for growth rings. Results indicate that saggital growth rings are laid down on a daily basis with the first daily growth ring laid down within one day of larval swim-up. Techniques for the preparation and reading of smallmouth bass saggitae are discussed. Examination of the saggitae of young-of-the-year smallmouth bass collected from 5 study sites within the New River drainage disclosed simultaneous spawning occurred during 1982, with a minor spawn of short duration occurring in late May and a major spawn of longer duration occurring during the last three weeks of June. ESTUARINF DEMERSAL FISH: A STUDY OF SPECIES DIVERSITY TRENDS IN A NEW ENGLAND FISH rOMMMUNITY. Mary G. Haoer* & A. J. Pulper, Jr.*, Dept, of Piol., Univ. of Va., Charlottesville, Va. 72901. Species diversity indices are frequently used to describe community structure. Classical studies comparing diversity trends have concluded that estuaries are characteristically species poor, a result of their exposure to extreme physiological stresses. These studies have been based primarily on sessile invertebrates. An examination of fish, however, has produced diversity indices that are quite high. Otter trawl samples of demersal fish in Long Island Sound and Narragansett Pay were taken during a five-day period in June 1982. Comparison of these samples with those of previous year-round studies of the re g. too suun;J- f'nt too estuarine fish community does not exhibit the predicted trend of low species diversity. Explanations for this phenomenon include the physiological and migratory capabilities of these species. THE VIRGINIA JOURNAL OF SCIENCE 123 EFFECTS OF HEXAVALENT CHROMIUM ON GROWTH AND REPRODUCTION OF CHIRONOMUS RIPARIUS (DIPTERA) . _C. L. Hax and A. L. Buikema , Jr., Biol. Dept., Virginia Polytechnic Institute and State Univ., Blacksburg, VA 24061. The effects of chromium (VI) on the complete life cycle of Chironomus riparius were examined in the labora¬ tory, using both a sublethal (0.5 mg/1) and a 50% lethal (2.0 mg/1) concentra¬ tion. Chironomus egg masses were introduced into both control and chromium treated 5-gal. aquaria. Larvae were subsampled at 5-day intervals, measured for length, and weighed to generate a growth curve. Emerging adults were allowed to mate and oviposit. Growth rates of larvae exposed to 0.5 mg/1 and 2.0 mg/1 were significantly higher than controls. Reproductive success, as measured by the number of offspring produced per adult female, declined with increasing chromium concentration. Adult weights and chromium body burdens in females did not dif¬ fer significantly between treatments and did not explain the observed decrease in reproductive success. EVALUATION OF SEAFOOD PROCESSING WASTES AS A PROTEIN SOURCE IN RAINBOW TROUT DIETS. L. A. Hel frich, J. C. Dean, D. L. Weigmann, and L. A. Nielsen. Dept, of Fisheries & Wildl. Sci., Virginia Polytech. Inst. & State Univ., Blacksburg, Va. 24061. Disposal of seafood processing wastes is a significant 1 problem for the seafood industry. In this study, we compared the growth response, survival, and feed conversion efficiency of cage-cultured rainbow trout (Salmo gairdneri) fed a control diet (Purina Trout Chow) with two experimental diets containing either herring (Clupea harengus) cannery residue or blue crab (Callinectes sapidus) scrap to evaluate the feasibility of using seafood wastes as dietary protein supplements in pelleted trout rations. One hundred rainbow tro^t finger! ings (40 g average) were stocked into each of nine i floating cages (1 nr) and fed one of the three diets of 3 % of live body weight per day for 179 days. Growth, production, survival, and feed conversion efficiency were significantly greater (p<0.05) for trout reared on the control diet than either of the experimental diets. Growth, production, and feed [conversion were significantly greater for trout reared on herring scrap than on the crab waste diet. Survival rates were similar for both experimental diets. AVIAN DISPERSERS OF EASTERN RED CEDAR ( JUNIPERUS V I RG INI ANA L.) AND THEIR INFLUENCE ON THE SPATIAL DISTRIBUTION OF THE RED CEDAR SEED SHADOW. A . M . A . Holthui j zen*and T.L.Sharik, Dept. Fish, and Wildl. Sci. and Dept. Biol., VPI & SU, Blacksburg, VA 24061. Eastern red cedar is mainly dispersed by birds, by passage of seeds through the gastro-intestinal tract. Eight species of birds were observed feeding on red cedar cones; yellow- rumped warblers, cedar waxwings, robins and starlings were the most commonly recorded species. Size of the red cedar fruitcrop was positively correlated with disperser abundance (no. dispersers/hour) . Disperser abundance was positively correlated with the numbers of seeds voided by birds and collected in the field, but not with red cedar fruitcrop depletion. Flock feeding ; species (e.g., cedar waxwings) caused rapid depletion of cedar fruitcrops. Red cedar dispersers can be divided into two groups: (1) consistent single foragers, providing local dissemination, and (2) flock feeders, occurring irregularly over time, which provide widespread long-distance dispersal. 124 THE VIRGINIA JOURNAL OF SCIENCE EASTERN RED CEDAR ( JUNIPERUS VIRGINIANA L.): AN EXAMPLE OF A PIONEER SPECIES WITH A HIGHLY EFFICIENT DISSEMINATION SYSTEM. A . M . A . Holthui j zen*and T.L.Sharik, Dept. Fish, and Wildl. Sci . and Dept. Biol., VPI & SU, Blacksburg, VA 24061. Seed dispersal theory predicts that pioneer woody species, producing fruits relatively high in carbohydrates and water, are mainly dispersed by opportunistic frugivores, which provide low quality dispersal. Low quality refers to (1) damage to the seeds by dispersers during ingestion or during passage through the gastro-intestinal tract, and (2) a large proportion of the fruitcrop wasted by dispersers. Red cedar produces large crops of small cones eaten by a variety of avian frugivores. Opportunistic frugivores, like the yellow-rumped , warbler, provided reliable dispersal and greatly enhanced germination of the seeds. The cedar waxwing, the only frugivores specialist, enhanced germination only slightly and provided unreliable dissemination. A large proportion of the fruitcrop of pioneer species is in general wasted. However, 73% of the red cedar fruitcrop was deposited beyond the canopy projection of the red cedar seed sources. EFFECTS OF ACCLIMATION TEMPERATURE AND ROPY SIZE ON THE METABOLIC PATE OF THE FRESHWATER CLAM, MUSCULIUM PARTUMEIUM (SAY) (PIVALVIA: PISIDJIDAE). P.J. Hornbach, Dept. Biol., Univ. Va., Charlottesville, Va. 22901. Metabolic rates (Vp?) were determined for 8 size-classes of clams for each of 16 treatments; Vp~ I measured at each of 4 run temperatures (RT = 5, 10, 15 & 20°C) for animals ac- i climated to 4 temperatures (AT = 5, 10, 15 &■ 20°C). For all treatments, except AT=5-PT=5, h-values from Vg_=aAEDW^ (AEDW=tissue weight) were not significantly different from 1, thus 0q IVg^/AFDW) is a constant for all sizes of clams. 0q varied from 0.40 -2.46 PlOp/mg/hr for treatments AT=10-PT=5 and AT=15-RT=2u respectively. AN0VA on these 15 treatments (all except AT=5-PT=5) indicated that both AT and RT, and their Interaction significantly affect Og^. Po«, when RT = 10 or 15, was higher for AT = 20 than AT = 5, 10, or 15 (reverse compensation); AT had no effect when PT=5 (no compensation). hlo's=1.0 over the interval 10-15°, regardless of AT, while Ojps were >1.0 over other intervals. The h-value, from Vg^aAEDW^1, for AT=5-PT=5 was 0.504, indicating that small clams had higher Og^s than large clams (e.g. 0g? = 2.19 and 0.22 for 0.03 & 3.00 mg clams respectively). Thus at low temperatures, size influences accli¬ mation pattern (small to large clams display the entire range of compensa¬ tory patterns; over to reverse compensation). BLOOD GLUCOSE LEVELS, DEPOT EAT, AND SURVIVAL TIME IN THE BILATERALLY ADRENALECTOMIZED MONGOLIAN GERBIL(MERI0NES UNGUICULATUS) . A. B. Jefferson* and F. B. Leftwich. Dept, of Biology, Univ. of Richmond, VA. , 23173 Forty Mongolian gerbils were bilaterally adrenalectomized and treated with injections of cortisol(l mg/2 days) and/or 5% sucrose drinking solution. Cortisol was found to cause a significant(p<0.05) increase in survival time of ADX gerbils(5.57 days vs. 3.57 days) and a significantly greater percentage loss of body weight over the course of the experiment, although the average percentage of weight lost per day was not significantly different for any group. Plasma glucose levels, measured on the third postoperative day, were significantly higher in ADX gerbils treated with cortisol than in untreated gerbils. No difference was found in either the weight of depot fat or the lipid content of depot fat under any treatment. The gerbils are not believed to have utilized any of the 5% sucrose solution, and sucrose was not found to have an effect on any parameter measured. THE VIRGINIA JOURNAL OF SCIENCE 125 ELECTROPHORETIC ANALYSIS OF HUMAN SERUM FOR CELL CULTURES. Evelyn W. Jemison and Angela D. Smith. Cytogenetics Laboratory, Department of Life Sciences, i Virginia State University, Petersburg, Virginia 23803. This investigation is designed to determine specifically the protein compo¬ sition of human serum fractions by electrophoretic analysis. Human serum was substituted as a growth factor for fetal calf serum (Jemison, Goad, and Robinson 1981) and was reported ineffective for the maintenance of lymphocyte growth cultures, thus, protocols were developed to identify the inhibitory proteins. The human serum fractions were produced by DEAE column chroma¬ tography and identified by the colored bands absorbed. Eluted fractions were electrophoresed on cellulose acetate and acrylamide gel for identification of proteins and protein subunits. We will report the electrophretic patterns of the human serum fractions. This research was supported by NIH/MBRS GRANT Number RR-08090-11 an NIH/MARC GRANT Number GM-07678. HUMAN SERUM FRACTIONS AS FETAL CALF SERUM SUBSTITUTE IN CELL CULTURE. Evelyn Jemison and Thomas Wright. Cytogenetics Laboratory, Department of Life Sciences, Virginis State University, Petersburg, Virginia 23803. Diethylaminoethyl (DEAE) cellulose extracted chromatographic fractions are being used as a fetal calf serum substitute in the lymphocyte test system. These experiments are designed to test for consistent effects wharf- human serum fractions of variable amounts are employed as growth factors. In order to determine the iri vitro effect, parameters being measured are growth rate, morphology and ploidy. Preliminary results suggest definitive cytotoxic effects when cells are exposed to human serum fractions. Exper¬ iments are in progress to more completely assess* the cytogenetic effects of the eluted serum fractions. This research was supported by NIH/MBRS GRANT Number RR-08090-11. MICRO-TITER ANALYSIS OF DNA SYNTHESIS OF HUMAN LYMPHOCYTES EXPOSED TO CALCIUM CYCLAMATE. Jemison, E. W., and Little, D. M., Department of Life Sciences, Virginia State University, Petersburg, Virginia 23803. Micro-titer analysis of DNA synthesis was used as a quantitative measurement along with the pro! i feration of PHA stimulated human T-lymphocytes in vi tro. After 24 hours (T?.) lymphocyte_cul tures were exposed to a calcium i cyclamate concentration of 1.0 X 10 D gm/rnl . Nine hours prior to cell harvest lymphocyte cultures were inoculated with tritiated thymidine (14.47 m ci/ml). The T7? lymphocytes were harvested on glass filter pads (mini -MASH) . Radiotracer activity of each miroculture was determined by 'scintillation counting. Data collected on lymphocyte cultures that were exposed to calcium cyclamate indicated a lower activity than the control. These results are consistent with those obtained in previous studies which used a macro system of determining the rate of DNA incorporation in cyclamate exposed cells. This research was supported by NIH/MBRS GRANT Number RR-08090-11. I 126 THE VIRGINIA JOURNAL OF SCIENCE SISTER CHROMATID EXCHANGE PATTERNS AS PRODUCED BY HUMAN SERUM FRACTIONS. Evelyn W. Jemison and Cornell H. Jones. Cytogenetics Laboratory, Depart¬ ment of Life Sciences, Virginia State University, Petersburg, Virginia 23803. This report is of an investigation designed to moniter cytological, chromosomal and growth rate changes in cells cultivated in human serum fractions. Fractions were eluted during DEAE. cellulose chromatography and substituted for fetal calf serum in the tissue culture growth medium. The sister chromatid exchange method is employed as a sensitive means of assessing chromosomal exchanges in vitro, which may be indicative of mut¬ agenic effects or toxic effects. Preliminary results have shown chroma¬ tid exchange in the presence of fraction one serum fraction. This report will also present data regarding proliferative rates and cell morphology. This research was supported by NIH/ MBRS GRANT Numer RR- 08090-11. EVALUATION OF OLIGOSPERMIC HUMAN SPERM WITH ZONA-FREE HAMSTER OVA. Keith Jones and R. James Swanson, Dept, of Biol. Sciences, Old Dominion Univ., Norfolk, VA 23508. Hamster ova, stripped of cumulus oopherous and zona pellucida by hyaluronidase and trypsin, respectively, have been used by a number of laboratories to evaluate the ability of human sperm to penetrate the oolemma. In this experiment, a group of suspected infertile, oligospermic sperm samples was compared to a group of proven fertile sperm samples. Nine specimens having a count below 5 x 10^ sperm/ml had a 4.7% mean penetration. Four samples falling between 5 to 20 x 10^ sperm/ml had an 18.8% mean penetration. Fifteen specimens having a count above 20 x 10^ sperm/ml had a 40.4% mean penetration. Severe oligospermia may have an adverse effect on sperm penetrability of the oolemma. EFFECTS OF WATERSHED URBANIZATION ON STREAM INSECT COMMUNITIES. R. Christian Jones and Christopher C. Clark*, Dept, of Biol., George Mason Univ., Fairfax, V a. 22030. The effect of watershed urbanization on stream insect communities was quantified by sampling streams draining thrity watersheds in northern Virginia ranging from highly urbanized Tripps and Holmes Runs to entirely forest Quantico Creek. Forest streams contained diverse fauna including species of Ephemeroptera , Trichoptera, Odonata, Diptera, Coleoptera, and Plecoptera. The most highly urbanized streams contained only Diptera, Trichoptera, and Ephemeroptera . Insect abundance was usually lowest in the most urbanized streams, except below reservoirs where Trichoptera were ex¬ tremely abundant. These results suggest that stresses resulting from water¬ shed urbanization deplete the stream insect fauna. EFFECT OF MILK REMOVAL ON TRANSFER OF PROLACTIN FROM BLOOD INTO MILK IN DAIRY COWS. Ray M. Kaplan* and R. Michael Akers, Dept, of Dairy Science, VPI&SU, Blacksburg, VA 24061. Three experiments were conducted to study the effect of milk secretion rate on prolactin (Prl) transport into milk. Foremilk, primary and residual milk samples (N=16) had similar Prl concentrations. Four cows were milked 2, 4, 8, and 16 hours after an initial milking. Milk¬ ing interval had no effect on Prl concentrations, however, total Prl trans¬ ported into milk (1.1 yg:2 h to 18.4 yg:16 h) increased with milk production. Prl was also infused IV (N=3) for 10 h to determine the effect of increased serum Prl on transport. Serum levels increased from 32 + 5 to 69 + 1 ng/ml . Concentrations of Prl in the milk, of active glands increased 82% by the end of infusion compared with 21% in unmilked mammary glands. Total Prl trans¬ ported during infusion was also greater in actively secreting glands (109 + 23 vs 45 + 10 yg). Results indicate that increased serum Prl has little effect on milk Prl in the absence of active milk secretion. THE VIRGINIA JOURNAL OF SCIENCE 127 THE ROLE OF DIFFERENTIAL VISIBILITY IN SIZE-SELECTIVE ZOOPLANKTIVORY BY BLUEGILL SUNFISH. William E. Kelso* and John J. Ney, Dept. Fish. Wild! . Sci . , VPI & SU, Blacksburg, V a. 24061. Size-selectivity by zooplankti vorous fish has fre¬ quently been reported and may exert a controlling influence on the composition of lentic zooplankton communities. Laboratory studies have shown that size- selection may be attributed to differential visibility of zooplankton to their predators, but this explanation has not been subjected to field verification. We collected juvenile specimens of the bluegill, a faculative planktivore, and zooplankton concurrently from Claytor Lake, Virginia, and compared length fre¬ quency distribution of zooplankton in gut contents and water column samples. Bluegills demonstrated size selection. Application of a differential visibility foraging model to the water column zooplankton size distribution failed to account for the observed prey selection. Other factors (zooplankton behavior and diversity, habitat complexity) appear to override visual discrimination in the selection of zooplankton by lentic bluegill. CHARACTERIZATION OF A SMALL ACCESSORY CHROMOSOME FOUND IN A NORMAL HUMAN FEMALE. Alice J. Kutterof f *and Stanton F. Hoegerman , Dept, of Biology, Col. of William and Mary, Williamsburg, Va. 23185. A small accessory chromosome was first found in a college student and later in her only sib (a sister) and in their mother. All are phenotypically normal and of high intelligence. The accessory appears to be monosatellited but silver staining revealed the presence of nucleolus organizer regions (NORs) on both arms. One NOR, which is probably located on the obviously satellited arm, is conspicuously larger than the second NOR. The accessory is probably monocentric as it contains very little, if any-, C-band positive chromatin. These observations argue for origin of the accessory chromosome by Robertsonian translocation. SOURCES OF VARIABILITY IN THE ACCUMULATION OF HEAVY METALS BY BLUEGILL SUNFISH AND CHANNEL CATFISH IN A VIRGINIA RESERVOIR. Mallory G. Martin* and John J. Ney, Dept, of Fish. Wild! . Sci., VPI & SU, Blacksburg, Va. 24061. Variability in reported trace metals concentrations of fishes is. high, but factors affecting bioaccumulation are poorly understood. We examined the relation of duration of exposure (age) to whole-body concentrations of Cd, Pb, and Zn in fishes from a metal -contaminated area of Claytor Lake, Virginia. We further reasoned that concentrations at age should be higher in benthic catfish than the more pelagic sunfish. Significant (P<0. 05) positive correlations were obtained between age and metals concentration (yg/g, D.W.) for Pb in both species; age-concentration correlations were significantly negative for Zn. Cd differed significantly but inconsistently among catfish age groups. Between species, catfish were signi¬ ficantly higher in Pb and lower in Zn than bluegills; no interspecific diff¬ erences were evident for Cd. EFFECT OF LEAF LEACHATES ON COLONIZATION OF ARTIFICIAL SUBSTRATES. Paul McCormick*, J.R. Pratt*, P.M. Stewart, and J. Cairns, Jr., Dept of Biol, and Ctr. for Environmental Studies, Va. Polytechnic Inst. & State Univ. , Blacksburg, Va. 24061. Secretions from aquatic macrophytes have inhibitory effects on phytoplankton, invertebrates, and macrophyte seedlings. Protozoan colonization of artificial substrates in Pandapas Pond, Montgomery Co., Va. was depressed near monospecific Typha latif olia stands relative to other areas of the pond. Microbial colonization was tested in laboratory microcosms filled with pasteur¬ ized leachates of Typha stems and Quercus leaves and a control of pasteurized water. Results of colonization trials indicated no significant differences in protozoan colonization; however, diatom colonization rates differed in each of the three treatments. Diatom colonization was depressed in Quercus leachates and elevated in Typha leachates. Fluorometric and spec trophotome trie analyses showed different levels of ring and double-bonded carbon compounds in the leach¬ ates. Results suggest that complex leachates may inhibit periphytic microbial species . 128 THE VIRGINIA JOURNAL OF SCIENCE GREAT HORNED OWL RESPONSE IN YOUNG VERSUS OLD-GROWTH FOREST STANDS. Kevin McGarigal & James Fraser, Dept, of Fisheries and Wildlife Sciences, V a. Poly¬ technic Inst. & State Univ., Blacksburg, VA 24061. To assess the impact of stand age and associated vegetation structure on great horned owl (Bubo virginianus) populations, we compared the frequency of owl response to tape recorded vocalizations in 24 young-growth (<80 years old) forest stands with the response frequency in 25 old-growth (>80 years old) forest stands. Stands were randomly selected from a list of all stands on the Blacksburg Ranger District of the Jefferson Nat. Forest. Owls responded at 5 of 25 old-growth stands and 1 of 24 young-growth stands (y2 = 2.86, 1 df, p = .09). The results indicated that more responses were obtained at stands adjacent to farmland. Therefore, we fit a model to the data using response and adjacency to farmland as variables. Responses were obtained at 5 of 17 stands within 1 k^i of farm¬ land compared to only 1 of 32 stands 'not within 1 km of farmland (y = 7.14, 1 df, p = .008). A 3-dimenstional model including both stand age and farmland variables, which assumed2all 2-way interactions but no 3-way interaction, did not fit the data well (Gz = 7.25, 1 df, p = .008) suggesting that owls occur more frequently where old-growth and farmland are juxtaposed than in other areas. INDUCTION OF ESTRUS IN WHITE-TAILED DEER. Kevin McGarigal & P. F. Scanlon, Dept, of Fisheries and Wildlife Sciences, Virginia Polytechnic Institute and State University, Blacksburg, V a. 24061. Attempts were made to induce estrus in adult female white-tailed deer (Odocoileus virginianus) prior to the normal breeding season (late November). Progestagen treatments were given for 10 days starting 11 October by i ntra-vaginal sponge-pessaries (medroxyprogesterone acetate, MAP, Upjohn, 60 mg) in 5 does or by subcutaneous silastic implants containing 75 mg melengestrol acetate (MGA, Upjohn) in 5 does. At progestagen withdrawal FSH (FSH-P Armour-Baldwin) was given i/m, 10 Armour units (AU) or 15 AU to does. All treatments were randomly allocated. Estrus was checked using 2 intact adult male deer to which the does were exposed every 12 hours from 20 to 80 hours after progestagen withdrawal. One pessary was lost by a doe. All 4 remaining pessary-treated deer elicited teasing responses by the males (2 were teased, 1 was the subject of persistent teasing, and one was bred by the male). Of the 5 implant-treated does only 1 elicited persistent teasing by the males--all others provoked no response by the males. Effectiveness of treatment in inducing estrus may have been limited by the timing and single-dose nature of the FSH injection and by the relative lack of interest of the male deer. NOTES ON THE BIOLOGY OF ALGOPHAGUS PENNSYLVANICUS, A MITE INHABITANT OF WATER- FILLED TREEHOLES. Donna L. Middleton & Norman J. Fashing, Dept, of Biol., Col. of William & Mary, Williamsburg, Va. 23185. A survey of water-filled treeholes in Cook Forest State Park, Pa., revealed that 54% contained Algophagus pennsylvanicus . A similar survey in southwestern and eastern Virginia, however, indicated a patchy distribution in this region with few treeholes containing this species. Investigations on feeding behavior were carried out by rearing mites on leaves as well as on various microbial isolates from treeholes (fungi, actinomycetes , bacteria). The ability to develop to adulthood and to reproduce was used as a measure of success. Mites were reared most successfully on fungi. Scanning electron microscopy of leaf surfaces on which mites had grazed revealed only small amounts of fungi when compared to controls, and SEM micrographs of mouthparts revealed adaptations for grazing. Therefore, A^_ pennsylvanicus feeds by grazing on fungi, in contrast to Naiadacarus arboricola , another mite inhabitant that feeds on decomposing leaves within the treeholes. THE VIRGINIA JOURNAL OF SCIENCE 129 ISDN-LETHAL PREDATION IN TEREERA DISLOCATA (GASTROPODA) . Ronald S. Mollick, Dept, of Biol, and Environ. Sci., Christopher Newport College, Newport News, Va. 23606. T. dislocata has been ccmnonly reported as a deposit feeder. In nature it was found concentrated in the casting piles of the hemichordate Balanoglossus aurantiacus . Laboratory studies (ATP, organic carbon) on casting piles and deposit feeding habits ruled out that mode of life. Food prefer¬ ence studies demonstrated that T. dislocat is a specialized predator feeding on the posterior sections of the hemichordate which typically loses only a fraction of it's body. Since the hemichordate can regenerate lost body parts it therefore is capable of providing a continuous yield of food. This rela¬ tionship was termed non-lethal predation which may play a significant role in the trophic dynamics of T. dislocata and other organisms. ULTRASTRUCTURE OF OCTOPUS MYOCARDIUM. Laura L. Monfalcone & C. P. Mangum, Dept, of Biol., Col. of William and Mary, Williamsburg, Va. 23185. Transmission elec¬ tron micrographs of the systemic ventricle of Octopus bimaculus reveal a cross- striated organization of the myocardial fiber. Sarcomeres are delineated by Z- bodies and filament bands, although A and I bands are not discernible. Mito¬ chondria are abundant and they contain many cristae. The extensive reticulum includes a t-tubul e-like system connecting subsarcolemmal vesicles to the inte¬ rior of the myofibrils along the level of the Z-bodies. No cell junctions were observed. An unusual component, a membrane bound muscle bundle, was found in sections from both the periphery and the interior of the tissue. This filament- dense bundle and its associated mitochondria stain much darker than the sur¬ rounding myocardial tissue, which is of the general type found throughout the ventricle. These results indicate that the fundamental features of the octopod myofiber resemble those found in other advanced cephalopods, which differ con¬ spicuously from the primitive Nautilus . SMALL MAMMAL TRAPPING SUCCESS IN "NO-TILL" SOYBEAN FIELDS. W. Edward Montz, Jr., Patrick F. Scanlon, Jack A. Cranford and Roy L. Kirkpatrick, Dept, of Fisheries & Wildlife Sciences, and DeDt. of Biol., Va. Polvtech. Inst, and State Univ., Blacksburg, VA 24061. Live-trapping was conducted in and around two soybean fields employing "no-till" farming practices in which soybeans were planted as a second crop in wheat stubble remaining from the previous crop. A total of 32 small mammals (30 Mus musculus, 1 Rattus norvegicus, 1 Peromyscus leucopus) were trapped over 574 trap-nights (5.6% overall trapping success) . Fourteen animals were trapped in 3 transects placed on field edges over 427 trap-nights effectively sampling 11,250 nr resulting in a mean estimated density of 11.8 animals/ha. Seventeen animals were trapped in two grids placed in field in¬ teriors over 144 trap-nights sampling 3,375 m^ producing an estimated mean den¬ sity of 50.5 animals/ha. Trapping success was significantly (P<0.0005, Chi- square = 42.4, 1 d.f) greater in field interiors than around edges. Differences in trapping success indicate that "no-tilled" fields may create favorable micro¬ habitats for small mammals by creating shade, by lowering ambient temperatures during warm summer growing seasons, by increasing food supply (insects, weed seeds, vegetation) and by enhancing escape cover. EFFECTS OF THE 0RGAN0PH0SPHATE INSECTICIDE, PARATHI0N, ON REPRODUCTIVE CHAR¬ ACTERISTICS IN Sylvilagus floridanus. W. Edward Montz, Jr. , Roy L. Kirkpatrick & Patrick F. Scanlon, Dept, of Fisheries & Wildlife Sci., Va. Polytech. Inst, and State Univ., Blacksburg, VA 24061. Cottontail rabbits, Sylvilagus floridanus , were live-trapped and placed in 0.10 ha enclosures in a deciduous woodlot. Treated female cottontail rabbits were given two 8 mg doses of ethyl parathion per kg body weight orally. No significant differences (P>0.05) were detected in fertility rate, litter size or weight, or weights of any of the internal organs measured when treatment groups were compared with controls. Peri-renal fat weights were significantly lower (P<0.05) in treated animals compared with controls. A subsequent laboratory experiment revealed that para¬ thion treatment caused animals to cease eating for 2 da following treatment. After this period of depressed food consumption , intake increased above that of controls suggesting compensatory feeding. 130 THE VIRGINIA JOURNAL OF SCIENCE EFFECTS OF FIELD APPLICATION OF THE CARBAMATE INSECTICIDE, METHOMYL, ON BRAIN ACETYLCHOLINESTERASE ACTIVITIES IN Mus musculus. W. Edward Montz, Jr., Patrick F. Scanlon, & Roy L. Kirkpatrick, Dept, of Fisheries and Wildlife Sci., V a. Polytech. Inst, and State Univ., Blacksburg, VA 24061. Free-ranging Mus musculus were live-trapped to determine the effect of Lannate (methomyl , I. 8EC, 2.3 1/ha) on brain acetylcholinesterase activity of field collected mice and to evaluate live-trapping as a technique for collecting insecticide-in¬ toxicated small mammals from soybean fields. Mean brain acetylcholinesterase activities over the 3 da collection period were significantly reduced (P<0.10) II. 2% below mean control activity. Maximal depression of 12.3% brain acetyl - cholinesterase activity occurred on the third day after spraying. Live-trapping is effective in collecting animals exhibiting brain acetylcholinesterase inhibition although a conservative sampling bias is probable. We conclude that measurable intoxication of field populations of small mammals may occur even when anticholinesterase insecticides are applied at recommended label rates and that M. musculus is a useful representative animal for investigations involving warm-blooded animals. (This study was supported, in part, by the Virginia Commission of Game and Inland Fisheries). FAT-SOLUBLE CHEMICAL RESIDUES IN BOBWHITE QUAIL FROM VIRGINIA. W. Edward Montz, Jr., Patrick F. Scanlon, Richard G. Oderwald, Roderick W. Young, Roy L. Kirkpatrick, and Jack V. Gwynn, Dept, of Fisheries & Wildlife Sciences, V a. Polytech. Inst. & State Univ., Blacksburg, V a. 24061. Bobwhite quail (Colinus virginianus) wings (N = 160, 80 adults, 80 juveniles from a total of 5 counties) recovered from hunters during the 1981/1982 hunting season in Virginia were tested for fat soluble organic chemicals. Fat was extracted from wings after feather removal and analyzed by gas chromatography procedures. Measurable quantities (range, ppm) of chemicals were found as follows: PCB's in 46 of 160 (0.1-2.55); total DDTs in 58 of 160 (0.05-6.12); Heptachlor in 98 of 160 (0.08-9.50); Heptachlor epoxide in 57 of 160 (0.02-3.59); Dieldrin in 75 of 160 (0.03-7.04); Dursban in 58 of 160 (0.12-16.32); HCB in 41 of 160 (0.01-3.38); Diazinon in 24 of 160 (0.01 - 2.93); BHC in 16 of 160 (0.05 - 4.13). Others found were Paarlon in 3, Lindane in 3, Chlordane in 5, Aldrin in 7, Mocap in 2, Toxaphene in 1 and Methoxychlor in 3. ANATOMICAL EVIDENCE FOR SYNCHRONIZATION OF FAST TOADFISH SWIMBLADDER MUSCLE. Paul J. Mosca* & Michael L. Fine4, Department of Biology, Virginia Commonwealth University, Richmond, Virginia 23284. Communicatory sounds are produced by rapid contraction of fast sonic muscles on the swimbladder of the oyster toadfish Ops anus tau. The muscle contracts in almost perfect synchrony, i.e. there is an 0.3-0. 5 msec difference in onset of an action potential between the cranial and caudal ends of the muscle over a length of 40-50 mm (Skoglund, 196l). We looked for anatomical evidence of this synchronization by examining the position of labeled neurons within the sonic motor nucleus (SMN) after retrograde transport of horseradish peroxidase injected into rostral, mid, and caudal regions of sonic muscle. Regardless of injection site, motor neurons throughout the rostral-caudal extent of the SMN were filled with reaction product. Differences between regional labeling pattern within the SMN are minor and inconsistent. We conclude that there is no obvious somatotopic projection of SMN neurons upon sonic muscle fibers, and therefore, that all the fibers in the muscle will be activated at approximately the same time. Supported by Virginia Commonwealth University biomedical-grant-in-aid. THE VIRGINIA JOURNAL OF SCIENCE 131 DEVELOPMENT OF THIN SECTIONING AND ACETATE PEEL TECHNIQUES FOR DETERMINING AGE AND GROWTH PARAMETERS OF FRESHWATER MUSSEL POPULATIONS. Steven N. Moyer & Richard J. Neves,* Dept, of Fish. & Wildl. Sci., V a. Polytech. Inst. & State Univ., Blacksburg, Va. 24061. Age and growth characteristics of freshwater mussels are determined almost exclusively from external growth checks on shells. However, shell erosion, environmentally-induced checks, and the obscurity of growth lines near shell margins usually result in only approximate ages for this faunal group. Two internal shell aging methods, thin sectioning and acetate peel techniques, were developed to obtain greater accuracy and precision in determining specimen ages. Thin sectioning makes use of a low speed, diamond-blade saw to cut 250 ym cross-sectional slices of shell which are mounted on slides and aged. The acetate peel method consists of 1) etching a cross-sectioned shell in HCL, 2) applying acetone and an acetate sheet to the cut surface, and 3) removing the acetate after the acetone has dried. Significant differences in age estimates between internal and external aging methods were noted in older age classes of specimens. A comparative evaluation of the various aging techniques is presented. EFFECTS OF PREGNANCY ON FOOD AND WATER CONSUMPTION OF NOHMOTENSIVE(WKY) AND SPON¬ TANEOUSLY HYPERTENSIVE(SHR) RATS. V.A.Mutchler and J.L.Hart, Dept, of Biol., George Mason U. SHR rats have a higher food-induced water intake, water/food ra- tio(w/F), than do WKY rats, which may contribute to the elevated blood pressure (BP) of the SHR rats0 During pregnancy, BP of SHR rats declines to normotensive levels. These studies determined if this BP drop in pregnancy was associated with a decrease in W/F. Food and water intake and systolic HP were measured in 6 WKY and 6 SHR rats before and at 1 wk intervals during pregnancy. The W/F(2.23+.l)of the NPG SHR rats and BP(274+8mmHg) were higher than those of the NPG WKY rats(l. 98+.1) (223+llmmHg) . During pregnancy, the BP of WKY rats remained at NPG levels until the 3rd wk wheiit was 183+10mmHg, while the W/F did not change. However, BP of SHR rats dropped to normotensive levels during the 1st wk and remained at this level; the W/F did not change during the 1st wk, but increased to 2.82+. 2 during the 2nd wk. These results suggest that factors other than a return to a noimoten- sive pattern of food-induced water intake contribute to the SHR BP drop in preg. ZOOGEOGRAPHY OF THE MUSSEL FAUNA IN THE NEW-KANAWHA RIVER. Richard J. Neves, Va. Coop. Fish. Res. Unit, Va. Polytech. Inst, and State Univ., Blacksburg, Va. 24061. The New-Kanawha River is considered to be the oldest river system in North America, and yet the aquatic fauna in the upper drainage (New) is noticeably depauperate. Two kilometers below the Gauley River and New River confluence in West Virginia is Kanawha Falls, one of the highest falls (7.3 m) in the eastern U.S. During the Pliocene and Pleistocene, the Teays River, precursor to the New and Kanawha Rivers, apparently provided a major corridor for mussel dispersal both across the Atlantic Divide and to adjacent rivers. Below Kanawha Falls, 34 mussel species have been reported, whereas only 12 species occur above the Falls. The inception of Kanawha Falls apparently had a significant impact on the dispersal of many aquatic groups. This paper reviews available data on the mussel fauna and discusses the probable effects of glaciation and biological barriers on mussel distribution. THE ROLE OF VISION AND OLFACTION IN PREY SELECTION BY THE SKINK SCINCELLA LATERALIS. Paul Nicoletto. Dept, of Biol., Va. Polytechnic Inst., Blacksburg, Va. 24061. The relative roles of vision and olfaction in prey selection by the lizard Scincella lateralis were investigated. The hypothesis tested was that S. lateralis would respond to prey (cockroach) using olfactory cues alone. Lizards were tested with four stimuli: visual, olfactory, concentrated olfactory, visual and olfactory, and a control. Re¬ sponsiveness to these stimuli was measured by the rate of tongue flicking during a five minute test period. The rate of tongue flicking to the olfactory stimulus alone was not significantly different from the control, but the concentrated olfactory, visual, and visual and olfactory were. Thus, response was primarily moti¬ vated by visual cues, except when the olfactory stimulus was strong. 132 THE VIRGINIA JOURNAL OF SCIENCE FOOD HABITS OF BROOK TROUT (SALVELINUS FONTINALIS) AND RAINBOW TROUT (SALMO GAIRDNERI ) IN ALLOPATRIC AND SYMPATRIC POPULATIONS. W. H. Nil sen* and L. A. Helfrich, Dept, of Fisheries & Wildl. Sci., Virginia Tech, Blacksburg, V a. 24061. Food habits of brook and rainbow trout in sympatric and allopatric populations were investigated to assess the extent to which trout species segregated into dietary niches. Collections of invertebrate drift, benthic invertebrates, and stomach contents of trout (110-210 mm) were made quarterly from August 1981 to June 1982 in three 200 m sections at Little Stony Creek, Giles County, V a. A total of 283 trout stomachs were examined. Stomach content analysis revealed that both species of trout were feeding more consistently on drifting than benthic invertebrates. Diet breadth of sympatric brook trout was generally narrower and exhibited greater seasonal variation than sympatric rainbow or allopatric brook and rainbow trout populations. Diet breadth of sympatric brook trout was narrowest when invertebrate resources were most abundant. Overlap values indicated that brook and rainbow trout utilized similar food resources. The lowest diet overlap values occurred when the density of benthic invertebrates was maximum and density of drifting invertebrates was high. These results suggest that diet segregation increases as food resources increase. AN EVALUATION OF ROCK BASS HABITAT IN VIRGINIA STREAMS. Paul Pajak*, Dept, of Fish, and Wildl. Sci., V a. Polytech. Inst. & State Univ., Blacksburg, V a. 24061. Reliable evaluations of fish habitat are critical for impact assessment and mitigative management. However, no evaluation techniques have escaped the operational constraints imposed by the practical limits of sampling, variable correlation/interaction, and consideration of unique on-site features. The Fish and Wildlife Service has recently developed a Habitat Evaluation Procedure (HEP) which attempts to describe habitat suitability for a particular species using Habitat Suitability Index models (HSI's) for essential habitat parameters. This procedure incorporates both habitat quality and quantity into "Habitat Units" (HU's) for the species being evaluated. HSI's have not been field validated for most species. This investigation sought to test the assumption that rock bass HU's and standing stocks were positively correlated. Results of fish marking studies suggest that temporal variation in habitat utilization may significantly affect model predictions based on a single "critical" evaluation period. INTERACTION OF THE ESTROUS CYCLE AND ADRENOCORTICAL ACTIVITY. Kathy Phillips* and Margaret Till, Dept, of Biol., Old Dominion Univiersity, Norfolk, VA 23508. Contradictory results have been found in changes in adrenocortical function dur¬ ing the estrous cycle. A glucocorticoid binding protein of adrenal origin has been established and its activity appeared to be correlated with adrenocortical activity. The present study was undertaken to further delineate the role of the estrous cycle in adrenal function and its relationship to glucocorticoid binding activity (GBA) . Female Sprague-Dawley rats (85-105 da) with normal estrous cycles were sacrificed at one of five designated stages: proestrus, early estrus, estrus, metestrus, and diestrus, and blood was collected. Adrenal glands were excised and incubated for 60 minutes. Media and plasma were analy¬ zed for corticosterone content and media were assayed for GBA. Plasma corti¬ costerone and secretion levels significantly greater during early estrus. GBA was significantly and negatively correlated with corticosterone secretion iri vi tro . THE VIRGINIA JOURNAL OF SCIENCE 133 CHROMATOGRAPHIC ANALYSES OF POLYCYCLIC AROMATIC HYDROCARBONS IN TISSUE EXTRACTS OF CONTAMINATED OYSTER POPULATIONS. Charles A. Pittinger and Arthur L. Buikema, Jr., Ctr. for Environmental Studies, Va. Tech, Blacksburg, Va. 24061. Indigen¬ ous oysters from contaminated locations in the lower James and Elizabeth Rivers, Va., were analyzed for the presence of nonsubs tituted polycyclic aromatic hydro¬ carbons (PAHs) . Extracts of freeze-dried samples were prepared by soxhlet re¬ fluxing in methylene chloride. PAH fractions isolated by gel permeation chro¬ matography (SX-8 Biobeads , 5 ml methylene chloride/min, 5 psi) and concentrated by rotary evaporation were analyzed by gas chromatography (6 ft. SP-2100 column, FID, 30 ml N2/min, 100-285° C. at 5°/min) . Concentrations of individual PAHs, including phenanthrene , fluoranthene, pyrene, chrysene, and benzo(a)pyrene , ranged from 0.027 to 2.9 ppm dry weight. Elizabeth River oysters contained greater numbers and higher concentrations of PAHs than did James River samples, presumably due to higher levels of fossil fuel contamination in the Elizabeth River . THE REDISCOVERY OF CAPILLARIA SERPENTINA (NEMATODA: TRICHUROIDEA) FROM FRESH¬ WATER TURTLES IN VIRGINIA. Thomas R. Platt, Dept, of Biol., Univ. of Richmond, Richmond, Va. 23173. Capillaria serpentina Harwood, 1932, was found in the mucosa of the small and large intestine of the snapping turtle, Chelydra serpen¬ tina, and the mucosa of the small and large intestine of Chrysemys j>. picta and Sternotherus odoratus , respectively, from Westhampton Lake in Richmond. Speci¬ mens compared favorably with previous descriptions of this nematode. Compari¬ sons of worms from different hosts and locations within hosts, in the present study, revealed no differences that would warrant establishment of new taxa. Cap . serpentina is, therefore, the only species of the genus reported from chelonians. This is the third report of Cap, serpentina and is a new state record for Virginia. CL picta and JS. odoratus are new host records. (Supported by a grant from the Univ. of Richmond Faculty Research Committee.) A HISTOLOGICAL EVALUATION OF THE EFFECTS OF A NON-PROTEIN DIET ON THE MOUSE THYMUS . F . V. Shef f er * , A. F . Conway , and P.L. Dementi, Dept. of Biol. ,Randolph-Macon Col., Ashland VA- 23005. The effect of a protein deficient diet on the thymus was investigated by quantitative histology. Mice were maintained on normal or protein deficient diets for durations of 1,2, 3, 4, or 5 days prior to age 35 days. At 35 days of age, mice were analyzed for thymus weight/gram body weight, % of thymus classified as cortex or medulla, and cells per field in the cortex or medulla. Control and experimental mice were compared using Student's t-test. Thymus weight was significantly reduced in experimental animals at all treatment times. Percent cortex was significantly reduced after 3,4, and 5 days of, treatment. Medullary cell density increased and cortical cell density decreased significantly over the treatment period. TISSUE EXPRESSION OF SELECTED ENZYMES IN THE AMERICAN SHAD, Alosa sapidissima. Edward N. Sismour, Steve Ackerman, and Ray S. Birdsong, Dept, of Biol., Old Dominion Univ., Norfolk, Va. 23508. The American shad (Alosa sapidissima), an important commercial and sport fish, has been declining in abundance. Mature adults segregate into populations at the time of spawning when they migrate from the ocean into natal rivers. It is during this spawning migration that adults, especially females, are subjected to intense fishing pressure. Knowl¬ edge of the genetics of natural populations is recognized as an important foundation for effective species management; however, little is known about the genetics of shad. A study in our laboratory is currently examining the allelic distribution of two enzymes, malate dehydrogenase and non-specific esterase, in several tissues from freshwater and mixed fishery stocks. Results of the study may elucidate means of differentiating populations of the American shad, which may be used for examining influences of various selective pressures on population structure. 134 THE VIRGINIA JOURNAL OF SCIENCE EARLY COLONIZATION PATTERNS OF PROTOZOANS AND DIATOMS. P.M. Stewart1, J.R. Pratt"*-', R.L. Lowe^*, and J. Cairns, Jr.-*-, -*-Dept. of Biol, and Ctr. for Environ¬ mental Studies, Va. Polytechnic Inst. & St. Univ., Blacksburg, VA 24061 and 2Dept. of Biol. Sci., Bowling Green St. Univ., Bowling Green, OH 43403. Coloniza¬ tion of artificial substrates by protozoans and diatoms was examined in 14 Michigan lakes and Pandapas Pond, Montgomery, Co., VA. Substrates were suspended in the littoral zone of all study sites and were harvested after 1, 3, 6, 15, and 21 days of exposure. The number of protozoan and diatom species were deter¬ mined. Results were fitted to the MacAr thur-Wilson model, S=Seq (l-e“Gt) , Esti¬ mates of colonization parameters were obtained by non-linear least squares regression. Equilibrium species numbers were similar, but not correlated, for both groups. The rate of species accrual was much greater for diatoms with equi¬ librium being attained in less than one day. Protozoan species equilibrium typically occured in about 7 days. Although these organisms are of similar size and share similar habitats, colonization dynamics appear vastly different. EFFECTS OF SPERM MOTILITY STIMULATORS ON CULTURED MOUSE EMBRYOS. Gordon K. Stokes , Steve Ackerman, and R. James Swanson, Andrology Laboratory, Dept, of Biol., Old Dominion Univ. , Norfolk, Va. 23508. Infertility, especially in oligospermic males, may be due to defective sperm motility. Recently, several substances have been studied for their abilities to stimulate sperm motility and/or ova penetration. The present investigation analyzed the effects of the following sperm stimulators on the growth of cultured mouse preimplantation embryos: arginine, caffeine, cysteine, dibutyryl cAMP, and taurine. Kinetics of embryo development in vitro were analyzed from the two-cell stage to the blastocyst stage over 72 hours. Preliminary evidence indicates that all sub¬ stances except arginine inhibit blastocyst formation. These studies may have implications for treatment of sperm used for either artificial insemination or in vitro fertilization. POPULATION DYNAMICS OF BLACK AND TURKEY VULTURES. T. M. Sweeney and J. D. Fraser. Dept, of Fisheries and Wildlife Sciences, VPI & SU, Blacksburg, VA 24061. The number of roosting black vultures (Coragyps atratus) and tur¬ key vultures (Cathartes aura) at a large roost near Radford, Virginia were counted as the birds left the roost at sunrise from 17 September 1981 to 31 March 1983. Counts were made at least three times monthly. Peak counts of roosting vultures were 498, 793 and 1107 on 21 September 1981, 5 December 1981 and 8 December 1982. The ratio of black to turkey vultures ranged from 1.6:1 on 10 December 1981 to 4.2:1 on 9 January 1983. Preliminary results suggest that annual fluctuations in numbers of roosting vultures are due to migration, changes in local food availability and use of seasonal roosts during the summer months. STUDIES OF GLYC0SAMIN0GLYCAN POPULATIONS IN CHICK LIMB DEVELOPMENT. Suzanne M. Thi em and Carolyn M. Conway, Dept, of Biol., Va. Commonwealth Univ., Richmond, Va. 23284. White Leghorn chick embryos were harvested at various stages during early limb development and fixed in 4% paraformaldehyde-O. 5% cetylpyri di urn chloride buffered at pH 7.2 Six micron paraffin sections were stained accord¬ ing to Alcian blue, PAS, or combined Alcian blue-PAS techniques in order to localize glycosaminoglycans (GAGs). PAS positive cell surfaces were found throughout the embryo. The extracel lular material between the ectodermal epi¬ thelium and underlying mesenchyme was alcianophi 1 ic. Heavier deposits of alcianophilic material were observed beneath the apical ectodermal ridge and in the region of the zone of polarizing activity of the developing limb. Bio¬ chemical characterization of the GAGs isolated from limb buds at various stages of development are in progress. THE VIRGINIA JOURNAL OF SCIENCE 135 CHARACTERIZATION OF A CORTICOSTERONE BINDING PROTEIN OF ADRENAL ORIGIN. Margaret Till, Dept, of Biol., Old Dominion Univ. , Norfolk, VA 23508. Adrenal incubation media possess glucocorticoid binding activity (GBA) . To investigate this phe¬ nomenon, adrenal glands were incubated for 15 minutes and subsequently for 60 minutes under basal or ACTH ( 200mU/ml) -stimulated conditions. Corticosterone secretion and GBA were significantly increased by ACTH. Gel electrophoresis of media showed the presence of seven proteins; three of which had GBA. Finally, adrenals were incubated as follows: basal conditions, ACTH (200mU/ml) stimula¬ tion, non-corticoid media components (NCMC) treatment, or ACTH and NCMC in com¬ bination. The NCMC was prepared by stripping media in which adrenals had been incubated of corticoids and adding this back to fresh media in concentrations equal to those following incubation. ACTH significantly increased corticoster¬ one secretion; while the NCMC treatment did not significantly change corticos¬ terone. In combination, ACTH failed to stimulate steroidogenesis, it appears that some non-steroid component blocked ACTH expression. 5PARTINA DECOMPOSITION BY INDIVIDUAL AND MIXED CULTURES OF SALT MARSH FUNGI. Albert P. Torzilli and George Andrykovitch, Dept, of Biol., George Mason Univ., Fairfax, Va . 22030. Cultures containing sterilized S parti na al terni flora tissue were inoculated with single or mixed inocula of three salt marsh fungi, Buergenerula spartinae, Phaeosphaeria typharum and Pleospora pelagica. After 42 days of incubation at 25°C, S parti na decom¬ position was compared in experimental treatments vs. uninoculated controls by determining the amount of residual total fiber, hemicell ul ose, cellulose and lignin. A one-way analysis of variance of these results indicated that all individual and mixed fungal combinations significantly degraded the total fiber, hemicel 1 ul osic and cellulosic fractions of Spartina tissue with the exception that, by itself, _P. typharum was unable to degrade cellulose. None of the individual or mixed inocula resulted in any lignin degradation. In most cases, mixed inocula resulted in a decreased ability of the competing fungal species to degrade the available S parti na substrate. COMPARISON OF PROTOZOAN COLONIZATION OF POLYURETHANE AND POLYESTER UNITS IN TWO LENTIC ECOSYSTEMS. Michael R. Van Brunt and William H. Yongue, Jr., Dept, of Biol., Va. Polytechnic Inst., Blacksburg, Va. 24061. Artificial substrate units of polyurethane foam(PUF) and of polyester fabric(PEF) were suspended in the littoral zone of oligotrophic Mountain Lake and low pH, soft water Pandapas Pond during summer 1982. Hexahedral PUF units (5x6 . 3x7 . 6 cm) and cylindrical rolled PEF units (h=10. 5 , d=5.5 cm) were used. Periodically, 3 units of each type were collected and the number of protozoan species present was determined, also water quality tests were performed on selected samples. Tests for nitrates, chlorides, pH, hardness, alkalinity and dissolved oxygen indicate the water in¬ side each substrate type is similar to the surrounding water in both systems. Of 16 possible fits (total species, flagellates, ciliates, sarcodines/per substrate/ per system) to the MacArthur-Wilson colonization equation (S (t)=Seq (l-e-^) ) lack of fit(°<=.05) was indicated in two cases, for flagellates on each substrate in Pandapas Pond. Between substrate types no significant differences in g-values were found, but two significant (©< =. 05) differences for Seq-values were found, for ciliates and sarcodines in Mt. Lake substrates. This data suggests that be¬ tween these substrate types colonization parameters are similar and that in some situations the species composition may be different. 136 THE VIRGINIA JOURNAL OF SCIENCE AN EVALUATION OF TECHNIQUES FOR KARYOTYPING MOUSE EMBRYOS. Allyson A. Van Steenbergen* and Carolyn M. Conway, Dept, of Biol., V a. Commonwealth Univ., Richmond, V a. 23284. Metaphase-arrested murine embryonic/fetal cells may be obtained following in utero exposure to Colcemid. Under conditions employed in our laboratory this method has not consistently produced adequate numbers of metaphase-arres ted cells for subsequent chromosomal analysis. Histological analysis was performed to determine if 9-11 day embryos had adequate numbers of mitotically active cells. Mitotic cells were observed in the majority of devel¬ oping organs at all three gestational ages0 Embryonic cells were cultured in vi tro in Eagle's Essential Medium supplemented with fetal calf serum and Colce¬ mid for two to 24 hours to determine if sufficient numbers of metaphase- arrested cells could be obtained. Shorter culture periods (2 hrs) produced the largest number of metaphase-arrested cells from 9 and 10 day embryos whereas longer culture periods were better for 11 day embryos. We have concluded that the in vitro culture technique is the appropriate technique for future studies. LIFE HISTORY ASPECTS OF THE REDLINE DARTER, ETHEOSTOMA RUFILINEATUM, IN THE NORTH FORK H0LST0N RIVER, VIRGINIA. James C. Widlak & Richard J. Neves, Va. Coop. Fish. Res. Unit, Va. Polytech. Inst, and State Univ., Blacksburg, Va. 24061. Life history aspects of the redline darter were investigated between May 1981 and May 1982 in the North Fork Holston River, southwestern Virginia. Analysis of scale samples indicated that males grew slightly faster and reached a greater maximum length than famales. Estimated annual survival for males and females was 0.44 and 0.33, respectively. Enumeration of stomach contents revealed that aquatic insect larvae were the major food items, dipterans predominating (67%-87%) at all times of the year. Peak feeding over a 24 hour period occurred from early to late afternoon (1600h-2000h) . Sex ratios favored males throughout the year, attributable to differential survival and distri¬ bution. Age I individuals of both sexes greater than 42 mm TL were sexually mature. Ripe males were collected in March although spawning colors first appeared in December. Ova development began in late February, and spawning occurred in riffle areas from mid-May to mid-August. AGGRESSIVE BEHAVIOR IN PEROMYSCUS LEUCOPUS AND P. MANICULATUS. Jerry 0. Wolff & Siobhan M. O'Connor, Dept, of Biol., U.Va., Charlottesville, VA, 22901, Raymond D. Dueser, Dept, of Envi. Sciences, U.Va., Ch'ville, VA, 22901, & Mark H. Freeberg, Dept, of Fisheries and Wildlife, Michigan State University, East Lansing, Michigan 48824. Aggressive behavior was studied in two sympatric species of mice, Peromyscus leucopus and P. maniculatus, to test for intra- and interspecific territoriality. Same sex mice were paired in intra- and interspecific behavioral trials conducted in the home range of one of the animals. Resident animals won from 53% to 93% of the encounters, regardless of the species of their opponents. In intersexual trials conducted within and between species, aggression occurred in less than 10% of the trials, and the behaviors of residents and intruders did not differ significantly within or between species. Since males and females show site-specific dominance against same sex intruders of either species and tolerate those of the opposite sex, there is no evidence of species recognition. THE VIRGINIA JOURNAL OF SCIENCE 137 COMPETITION BETWEEN PER0MY50US LEUCOPUS and P. MANICULATUS. Jerry 0. Wolff, Dept, of Piol., Lhiv. of Virginia, Charlottesville, Va, 22901 & Raymond 0. Dueser, Dept, of Environ. Sciences, Univ. of Virginia, Charlottesville, Va, 22901. A study was conducted to determine the ecological relationships between Peromyscus leucopus novehoracensis and P. maniculatus nuhiterrae which co-occur in the montane forests of southwestern Virginia. These two species are morphologically, ecologically, and hehaviorally similar and exhibit roughly synchronous population fluctuations. A stomach contents analysis showed that the two species have similar food hahits and a vegetation analysis revealed no significant microhahitat differences. In reciprocal removal experiments, the remaining species colonized the habitat vacated by the removed species. In both cases, the remaining species exhibited higher population densities on treatment grids than on controls. Patterns of space use showed that males and females of both species maintain exclusive home ranges within each sex and appear to exhibit intra- and interspecific territoriality. SPECIES RECOGNITION AND BEHAVIORAL ISOLATION IN PEROMYSCUS LEUCOPUS and P. MANICULATUS. Jerry 0. Wolff and Cynthia Van Cleef, Dept of Biology, Univ. of Virginia, Charlottesville, VA. 22901. Wolff et al. (1983) have shown that Peromyscus leucopus and P. maniculatus are intra- and interspecifically territorial within each sex. Wolff et al. (in prep) have shown that males and females are amicable toward each other regardless of species, exhibiting no discernible differences in agonistic behavior. The obj'ective of this research was to test for behavioral isolation and species recognition by conducting discrimination trials in the laboratory. In intersexual trials, males and females showed a preference for their own species, however, this was more pronounced in females than males. In intrasexual trials, males and females did not discriminate between species. We conclude that P. leucopus and P. maniculatus can distinguish between members of the opposite sex and are thus reproductively isolated. We were not however, able to demonstrate species recognition within sexes. The implications of this will be discussed. IDENTIFICATION OF AGING RELATED CHANGES IN THE REPRODUCTIVE TRACTS OF CD-I MICE. Monette W. Wood* and Carolyn M. Conway, Dept, of Biol., Va. Commonwealth Univ., Richmond, Va. 23284. Normally cycling female CD-I mice were grouped according to age (3, 6, 9, 12+ months) and breeding history (nul 1 iparous , multiparous). Within each age group females were sacrificed at different stages of the estrus cycle and their reproductive tracts (ovaries, oviducts, uterus, vagina) removed and weighed. Uterine tubes were flushed with physiological saline and the resulting irrigates used to determine the protein content of the intra-uterine tubal fluid (IUTF). Younger (3 month) nulliparous females showed the most distinctive weight change pattern (pre-estrus increase, post-estrus decline). In older nulliparous and multiparous females the relative reproductive tract weights did not show distinct changes during the estrus cycle. In recently bred females the relative reproductive tract weights continued to increase following estrus. The protein content of the IUTF in most age groups generally increased prior to estrus and then declined. 138 Botany Section THE VIRGINIA JOURNAL OF SCIENCE COMPOSITION, STRUCTURE, AND DYNAMICS OF THE SPRUCE-FIR FOREST ON THE SUMMIT OF MOUNT ROGERS. H . S . Adams . D. S. Lancaster Cmnty. Col., Clifton Forge, VA 24422 & S. L. Stephenson, Dept, of Biology, Fairmont State Col., Fairmont, WV 26554. The red spruce-Fraser fir (Picea rubens-Abies fraseri) forest on the summit of Mount Rogers in southwestern Virginia was intensively sampled to obtain data on composition, structure, and dynamics of vegetation of the north¬ ernmost extension of this southern Appalachian forest type. In addition, data were obtained on soil chemical and physical characteristics. Five tree species were present, but fir dominated all size classes with spruce the only important associate. Oxalis acetosella and Dryopteris campyloptera made up more than 95% of the total herb cover. The shrub stratum was poorly developed with just three species represented. This community is ecologically similar to other southern Appalachian spruce-fir forests. NATURE TRAIL DEVELOPMENT IN THESE TIMES OF BUDGET CUTS AND VANDALISM. Marion C. Blois , Div. of Natl, and Appl. Sci., Northern Va. Cmnty. Col . -Manassas Campus, Manassas, Va. 22110. Budget cuts, higher rates of vandalism, and an increasing need for community support for higher education are ail problems to be dealt with in all aspects of higher education. In developing and maintaining a Nature Trail, these issues must all be considered. In this presentation, measures to economize in trail development such as making one’s own signs for identification of plant species, use of student help, and use of locally available materials will t>e discussed. Suggestions for minimizing vandalism such as placing iden¬ tification signs on larger trees out of reach of teenagers and embedding cedar posts in the ground will be a second point of consideration. Finally, encour¬ agement of community use of a nature trail to build community support and inter¬ est in college activities will be briefly discussed. Appropriate slides will illustrate implementation of these ideas on the Manassas Campus Nature Trail. FLOWERING PLANT RECORDS FROM NORTHERN VIRGINIA. Ted R. Bradley, Dept, of Biol., George Mason Univ., Fairfax, V a. 22030. Recent collections of flower¬ ing plants from northern Virginia have revealed several unusual records. Species spreading from cultivation and apparently new to the state are Anagallis 1 ini folia ( Primulaceae) , Linaria maroccana (Scrophulariaceae) , Euonymus fortunei (Cel as traceae) , and Hosta ventricosa (Liliaceae). Ulmus pumila (Ulmaceae) is reported for five new counties. Dipsacus laciniatus ( Di psacaceae) is reported from Loudoun Co. and Kochia scoparia ( Chenopodiceae) from four new counties. Polygonum perfoliatum ( Polygonaceae) has reached Fairfax Co. from Maryland. WOOD ANATOMY AND RELATIONSHIPS OF THE HAWAIIAN ENDEMIC GENUS NEOWAWRAEA (EU- PHORBIACEAE) . Dorthe Brandt* & W. John Hayden, Dept, of Biology, Univ. of Rich¬ mond, Richmond, VA. 23173. Neowawraea was originally described as a distinct monotypic genus of the euphorbiaceous subfamily Phyllanthoideae . However, it has been generally treated as a species of the pantropical genus Drypetes . Three wood specimens of Neowawraea were studied for comparison with diverse genera of Phyllanthoideae. Anatomically, Neowawraea possesses simple perfora¬ tions, thin-walled septate fibers, and poorly developed axial xylem parenchyma, whereas Drypetes has scalariform perforations, thick-walled non-septate fibers, and abundant diffuse and diffuse- in-aggregates axial xylem parenchyma. Thus, wood of Neowawraea is inconsistent with a close relationship with Drypetes . On the other hand, wood of Neowawraea conforms closely with the "Glochidion-type" wood structure of Phyllanthoideae; its closest relatives are thus to be found among genera such as Glochidion, Breynia , and Phyllanthus , rather than .Drypetes . THE VIRGINIA JOURNAL OF SCIENCE 139 Host Range and Germination of Branched Brommrape , Orphan che ramos a L . , Texas Strain. Robert D . Coffin * and Lytton J. Musselman *, Dept, of Biol., Old Dominion Univ., Norfolk, Va. 23508 & Robert E. Eplee *, USDA, APHIS-PPQ, Whiteville Methods Development Center, Whiteville , N.C. 28472. In this green¬ house test the Texas strain of Orob anche ramos a parasitized tomato, tobacco, eggplant, bell pepper, sunflower, celery and bell bean. Tomato and tobacco supported emergent growth of the parasite; only the underground stages were observed on the other hosts. Optimal germination in vitro was obtained by conditioning and germinating seed at 18° C . The effectiveness of strigol and four strigol analogs, GR-7, GR-24, GR-42 and GR-60 to induce germination was tested in vitro . Highest germination was obtained with GR-24, GR-7 and strigol. Of the three compounds, GR-24 induced greater germination. Concentrations of 0.01 ppm GR-24 and 1.0 ppm GR-7 induced 75.8 and 73.8 % germination respect¬ ively. Further increases in concentration did not significantly change the germination response. Due to regulatory considerations, field evaluations were not included in this study. PRELIMINARY REPORT ON A LONG-TERM STUDY OF LOG DECAY IN THE GREAT DISMAL SWAMP. Frank P. Day, Jr. , Dept, of Biol. Sci. , Old Dominion Univ., Norfolk, Va. 23508 The progress of decay, weight loss, and nutrient dynamics are being monitored in preweighed red maple logs which were set out fresh on four sites with differ¬ ent hydroperiods. Early results show that after one year logs on the wet maple-gum site decayed more rapidly (21% weight loss) than on the other sites (<11% weight loss). Possible explanations for the site differences are dis¬ cussed. A preliminary conclusion is that under certain conditions, log de¬ composition may be much faster than previously estimated in the literature. WOOD ANATOMY OF BETULA UBER, THE VIRGINIA ROUND-LEAF BIRCH. W. John Hayden, Dept, of Biology, Univ. of Richmond, Richmond, Va. 23173, & Sheila M. Hayden.* Wood structure of Betula uber is described as a contribution towards an under¬ standing of the relationships of this rare and endangered birch. Salient features of Betula uber wood include: diffuse distribution of mostly solitary pores; vessel elements with scalariforxn perforation plates; minute, alternate intervascular pits; fiber- tracheids , often with gelatinous walls in the speci¬ men examined; numerous, non-aggregate, homocell ular procumbent rays; and pre¬ dominantly diffuse apotracheal axial xylem parenchyma. Wood of Betula uber is typical of birches in general and matches in detail many of the diagnostic characteristics of birch woods belonging to series Costatae ; however, it diverges strongly from woods of series Humiles. This conclusion is further supported by quantitative features of vessel element length, diameter, and frequency per cross-sectional area of wood surface. Relationship between Betula uber and species of series Costatae is thus favored over series Humiles . RIPARIAN VEGETATION DISTRIBUTION RELATIVE TO FLUVIAL LANDFORMS AND FLOOD FREQUENCY ALONG THREE NORTHERN VIRGINIA STREAMS. Cliff R. Hupp*, U.S. Geological [Survey, 431 National Center, Reston, Va. 22092. Persistent patterns of woody [vegetation distribution within the bottomland forest of three northern Virginia ;str earns were identified and related to fluvial land forms above the stream chan¬ nel. Vegetation patterns were investigated in valley bottoms of widely different sizes and related to channel geometry, streamflow characteristics, and sediment- size characteristics. Distinct vegetation patterns were found on four common fluvial geomorphic features: deposit ional bar, active-channel shelf, flood plain, and terrace. Vegetation data from these features were analyzed by binary [discriminant and principal components analyses. Results and related field obser¬ vations suggest that certain species are significantly associated with certain geomorphic features and that vegetation patterns develop as a result of particu¬ lar hydrologic processes that shape each geomorphic feature rather than from pediment-size characteristics or depth to water table. Flood disturbance appears jto be an important factor in maintaining the species patterns, which potentially plight be used as indicators for particular hydrogeomorphic conditions. 140 THE VIRGINIA JOURNAL OF SCIENCE VASCULAR FLORA OF EASTERN PRINCE WILLIAM COUNTY, VIRGINIA. Richard L. Keyser*, Dept, of Biol., Col. of William & Mary, Williamsburg, Va. 23135. Prince William County east of Interstate 95 was visited 21 times in 1981-82 with 526 species of vascular plants recorded. Seventeen of these, including Celosia argentea. Cannabis sativa and Datura met elo ides were strays or long-persistent from cult¬ ivation? Of the remaining 509 species, 119 (23$) were introduced but establish¬ ed. Despite much previous collecting, 96 of the 509 were new county records and Diervilla lonicera also a new Coastal Plain record. However 54 of the new spec- ies are introduced, reflecting the increasing disturbance of the area. The study area is twelve miles long and two to four wide, entirely within the Inn¬ er Coastal Plain, but dissected by six watersheds of the Potomac River, with a maximum relief of 222 feet. Tidal marshes had extensive emergent beds of Nuphar luteum . Rhus vemlx and Dryopteris cristata were rare members t>f the bottomland community. Distinct spring, summer and ±all weedy floras dominated disturbed areas. Deciduous woods and Pinus virginiana stands were other major habitats. INTRASPECIFIC VARIATIONS IN PROTEIN COMPLEMENTS IN ISOLATES OF ENDOTHIA RADICALIS . J. A. Micales and R. J. Stipes, Dept. Plant Pathology and Physiol¬ ogy, Va. Polytech. Inst, and State Univ. , Blacksburg, VA 24061. The identification of isolates of Endothia radicalis (=E. fluens) is difficult because of the morphological similarity and shared host range of it with E. parasitica. Vertical slab gel polyacrylamide electrophoresis was used to distinguish six monoconidial isolates of EL_ radicalis from two representative monoconidial isolates of JE^ parasitica. The protein profiles of the E. radicalis isolates differed from those of EL_ parasitica both in the number and position of the protein bands. The banding patterns of Greek isolates of E. radicalis were quite similar to that of a European isolate of this fungus (CBS 240.54), while the protein profile of a New Zealand isolate was dissimilar to those of the other E. radicalis and' E. parasitica isolates; this New Zealand isolate may represent a third species of Endothia. Vertical slab gel electrophoresis can be used to differentiate EL_ radicalis from EL_ parasitica, and to confirm or deny questionable identifications of isolates of these species . TAXONOMIC IMPLICATIONS OF COMPARATIVE SENSITIVITIES OF ENDOTHIA. PARASITICA AND EL GYROSA TO CAPTAN, CHLORTETRACYCLINE, DICHLORAN, FUNGISOL® 'AND VINCLOZOLIN. J. A. MICALES and R. J. Stipes, Dept. Plant Pathology and Physiology, Virginia Polytechnic Inst, and State Univ., Blacksburg, VA 24061. The differentiation of Endothia parasitica from EL_ gyrosa based on their ^elective sensitivities to the fungitoxicants cycloheximide and Arbotect 20-S has been previously demonstrated. This work has been expanded to test for further differential sensitivities of these species to the following compounds: Fungisol , chlor- tetracycline, vinclozolin, captan and dichloran. These compounds are represent¬ ative of the benzimidazole, tetracycline, substituted aromatic and dicarboxi- mide classes of fungitoxicants, respectively. The fungitoxicants were incor¬ porated into molten Difco potato-dextrose agar at increasing concentrations between 0.1 and 100 yg/ml. Mycelial plugs of five to ten monoconidial isolates each of EL_ parasitica and EL_ gyrosa were used as inoculum. After 8 days at 25 C, the radial growth of each colony and the percent growth inhibition based on the radial growth of nonamended control plates were determined. No significant differences in sensitivities to these compounds were observed. Endothia parasitica and EL_ gyrosa could not be differentiated by their growth responses to these compounds. THE VIRGINIA JOURNAL OF SCIENCE 141 APPLICATIONS OF REMOTE SENSING DATA TO EOTANICAL AND GEOBOTANICAL STUDIES IN THE VIRGINIA PIEDMONT. _N.M. Milton and M.D. Krohn, U.S. Geological Survey, Res ton, Va 22092. The Virginia Piedmont constitutes an ideal site for testing remotely sensed data from a variety of sources for botanical and geobotanical work. Remote identification of plant species has already proved useful as an aid for lithologic mapping. Data sets currently available include Landsat MSS and TM, SMIRR data from the second Columbia flight, and several radar data sets of differing wavelengths. Computer processing is essential for enhancing subtle differences in spectral reflectance or backscatter. So far we have 1) enhanced winter Landsat MSS data to identify chestnut-oak communities, which are known to occur on certain lithologies; 2) identified two pine communities on a density-sliced and color-coded radar image; 3) distinguished three pine communities on Landsat TM data; 4) used SMIRR data to extract spectral curves for plant communities of known composition for use in the interpretation of data from other sources, particularly from future Landsat TM data. POLLEN VECTORS OF WEST AFRICAN STRIGA HERMONTHICA . L.J. Musselman, Dept, of Biol. Sci. , Old. Dominion Univ., Norfolk, Va. 23507* E.C. Matteson,* Consortium for International Crop Protection, Berkeley, Ca. 94704, & S.J. Fortune, * Dept, of Biol. Sci., Old Dominion Univ,, Norfolk, Va. 23507. Striga hermonthica is a root parasite that causes serious yield reduction in subsistence grain crops in sub-Saharan Africa. Ibis parasite is strongly allogamous and reproduces ex¬ clusively from seed, but little information is available on factors involved in seed production. Floral visitors of S. hermonthica were collected from various sites in Upper Volta. These were examined for the presence of pollen using acetolysis and SEM microphotography. The results of this study indicate that S. hermonthica is visited by diverse insects and is probably not dependent on any single group for pollination, although considerations of floral morphology and length of mouthparts of insect visitors point to long tongued lepidopterans as likely being the most effective pollinators. TREE RINGS IN THREE DIMENSIONS. Richard L. Phipps. U.S. Geological Survey, Reston, VA. 22092. Standard techniques of dendrochronology utilize measure¬ ments from one dimension (radius at breast height) of a three-dimensional subject (tree trunk). The shape of Individual rings is near-paraboloidal, displaying a similarity in growth time trend between heights. Certain devia¬ tions In the expected growth trend appear explicable as functions of species and environmental conditions. The bulk of the annual growth increment in the upper part of the trunk takes place earlier in the growth season, and thus responds to different environmental conditions, than growth in the lower trunk. Measurements of rings at more than one height allow estimates of year- to-year height growth, and to seasonal variations in environmental conditions. Understanding i nterrel ationshi ps between heights is of potential value to Interpreting the growth trend at breast height. DISCRIMINATION OF VEGETATION PATTERNS IN WESTERN RANGELANDS BY MEANS OF REMOTE SENSING DATA. M.S_. Power*, N.M. Milton, M.D. Krohn*, and M.H. Podwysocki*, U.S. Geological Survey, Reston, VA 22092. Vegetation patterns, visible in remote sensing images, may provide information about soils, geomorphology, water availability, and mineralized areas. Remote sensing data acquired from radar and Landsat yield different geobotanical information. Radar images of the Escalante Desert in Utah enhance tonal features associated with discrete shrub communities, each growing on a different soil type. The tonal differences are related to shrub heights and canopy structure. Rangeland vegetation in the Pioneer Creek and Carpenter Creek drainages in Montana is closely tied to geomorphology. A Landsat compound ratio image enhances differences in spectral response that represent boundaries between vegetation communities growing on Tertiary lakebed deposits, lower slopes of alluvial fans, and upper slopes of alluvial fans. The Tertiary lakebed deposits may prove to be the best source for placer gold deposits. 142 THE VIRGINIA JOURNAL OF SCIENCE SPRING TO SUMMER TRANSITIONS IN PHYTOPLANKTON POPULATIONS OF THE U.S. NORTH¬ EASTERN CONTINENTAL SHELF. D. A. Randolph*, P. Zimba*, L. Jugan* , and H. G. Marshall. Department of Biological Sciences, Old Dominion University, Norfolk, Va. 23508. Phytoplankton results from 292 surface water samples, obtained during four 1982 cruises, indicated specific composition changes occurred from spring through fall across the northeastern shelf. Regional divisions of the shelf into north and south coastal, mid-shelf, and outer shelf stations showed each section to have a seasonally distinct assemblage, with a progressive succes- sional pattern. Late spring dinof lagellates and small diatom dominants were generally replaced by large sized dinof lagellates and diatoms in summer, with a mixed group common in fall. Several ubiquitous species were also found over the entire shelf. Regionally, dinof lagellates and small sized diatoms pre¬ dominated near shore and along the outer shelf, with larger species abundant at mid-shelf. Cryptomonads, cyanophyceans , euglenoids, and an ultraplankton component were also common over the shelf. Supported in part by NOAA/NEMP Ocean Pulse. STEM ONTOGENY IN CHAMAESYCE HIRTA (EUPHORBIACEAE) . Marilyn Rosengarten* & W. John Hayden, Dept, of Biology, Univ. of Richmond, Richmond, Va. 23173. Chamaesyce hirta is a weedy tropical species with a growth habit somewhat inter¬ mediate between erect and densely matted prostrate species in the genus. Stem ontogeny was studied from serial paraffin sections of early developmental stages of seedlings. As in all species of Chamaesyce, the epicotyl produces only a single pair of leaves and develops no further; growth resumes through initiation of lateral branch axes, the first pair of which are axillary to the cotyledons. The course of leaf traces, sites of subsequent lateral axis initiation, and vascularization of lateral axes are described in detail through various develop¬ mental stages. These ontogenetic data are significant in assessing Roeper's hypothesis concerning the phylogenetic derivation of Chamaesyce from Euphorbia. THE CURRENT STATUS OF VIRGINIA ROUND-LEAF BIRCH, BETULA UBER. T. L. Sharik, D. M. Porter, P. P. Feret, R. JE. Kreh, Depts. of Biology and Forestry, Va. Polytechnic Inst., Blacksburg, Va. 24061 and R. H. Ford, Southwest Virginia Community College, Richlands, Va. 24641. From 1975 to early 1982 the only known wild population of Betula uber , located in southwest Virginia, declined from 40 to 12 individuals. Clearing of forests, removal of litter, and scarification of soil in the winter of 1981-82 adjacent to the remaining reproductively mature trees resulted in the germination and establishment of 80 seedlings in the summer of 1982. This treatment, designed to simulate natural disturbance, indicated that 13. uber is capable of reproducing itself in the wild given the proper environmental conditions. The concurrent production of approximately 100 seedlings in the greenhouse from open- pollinated seed of EL uber will be used to establish several experimental populations on public lands within the next year. HOLOCENE (8000-6000 B.P.) FRUITS AND SEEDS FROM MACON COUNTY, ALABAMA. Donna j_. Shi pp, Dept, of Biol., George Mason Univ., Fairfax, Va . 22030. Holocene clays along Uphapee Creek were sampled for macroflora and pollen analyses on a U. S. Geological Survey sponsored field trip. One deposit was particularly rich in seed and fruit remains. Radiocarbon dating indicates an age of 6360 + 110 y B.P. (U.S. Geological Survey Radiocarbon-41, Reston, Va . ) . Seeds and fruits were isolated and examined with light and scanning electron microscopy. Seed and fruit genera found include Betul a , Rhynchospora , Fagus , Panicum, Ceratophyl 1 urn, Vi ti s , Parthenoci ssus , Quercus, Ca rya , Pi nus , Strophostyl es , Fraxinus , Liriodendron and Xanthium . Among these genera are aquatics and understory plants that have not been found in leaf and pollen remains. These genera indicate a flora similar to the flora present in the region today. THE VIRGINIA JOURNAL OF SCIENCE 143 MICROENVIRONMENTAL PARAMETERS OF FIVE UPLAND FOREST TYPES IN THE MOUNTAIN LAKE AREA OF SOUTHWESTERN VIRGINIA. S. L. Stephenson, Dept, of Biol., Fairmont State College, Fairmont, W. Va. 26554, and A. A. Normandy*, Dept, of Biol., George Mason Univ. , Fairfax, Va. 22030. Microenvironmental parameters of five upland forest types in the Mountain Lake area of southwestern Virginia were examined in conjunction with an investigation of the distribution and ecology of Myxomycetes in these same communities. Topographic parameters were determined and quanti¬ tative data on the structure and composition of the vegetation were obtained for a 0.1 ha study area at each site. Soil temperature, air temperature, and soil moisture were monitored during the period of June 21 to August 14, 1982. Soils associated with each site were analyzed for texture, organic matter, pH, Ca, P, K, Mg, Zn , Mn , NO3-N, and soluble salts. Other microenvironmental parameters measured or determined included light intensity, bark pH, and pH of the litter layer. The results of this study are described and discussed. (Supported by a Univ. of Va. post-doctoral fellowship and a undergraduate research scholarship.) DIATOM COMMUNITY ANALYSIS OF GRAVEL PIT PONDS. P.M. Stewart, Dept, of Biol., Va. Polytechnic Inst. & St. Univ., Blacksburg, VA 24061 and M.C. Miller5'*, Dept, of Biol. Sci., Univ. Cincinati, Cincinati, OH 45221. This study was designed to determine diatom community structure of gravel pit ponds and to examine environ¬ mental parameters that affect diatom assemblages. Samplers were placed in 33 gravel pit ponds near Cincinati, OH. After a three week colonization period, 250 diatom taxa were recorded, with 20-73 species/pond. Various null hypotheses were tested for possible explanations of diatom community composition. The number of species in a pond, the number of ponds in which a species was found, and the number of species shared between ponds were compared to neutral models and found to not be random. The MacArthur-Wilson model was rejected as a possible explana¬ tion of the number of species found in a pond. No distance effect on community similarity was observed. Correlational procedures utilized suggested a possible relationship between several diversity indices and environmental parameters, including substrate and productivity. THE MIRAGE OF PLANT HEALTH. R. Jay Stipes and Jessie A . Micales , Dept, of Plant Pathology & Physiology, Va. Polytech. Inst. & State Univ., Blacksburg, VA 24061 After the query, "What is this plant?," probably comes the next most often asked question, "What is wrong with my plant?" The charge given to plant path¬ ologists is to address the latter query. To understand the parameters of "dis¬ ease," one must have delimited those of "ease" or "health," since disease is considered to be an unhealthy state. To understand health, then, one must have a comprehensive grasp of the "normal" or "perfect" physiology , anatomy, morph¬ ology, ecology and other aspects of the patient in question. Since there is no plant for which all biochemical constituents and physiological reactions and their normal parameters are comprehensively known, then we must be using arbitrary absolutes and flexible parameters when we indicate that a plant is healthy or diseased . Perhaps in our experimentation and other considerations , we should refer to healthy plants as "apparently healthy" or simply as "control" plants. These concepts are especially germane to our diagnoses and measure¬ ments, for example, of abiotic stresses and disease complexes. In summary, absolute health and disease are on a continuum, with nebulous lines of demarc- action from a physiological perspective, but not from an economic one! 144 THE VIRGINIA JOURNAL OF SCIENCE RADIAL GROWTJ RESPONSES OF TWO TREE SPECIES TO CLIMATE IN NORTHERN VIRGINIA. J. C. Wh I ton , U. S. Geol. Survey, Reston, Va. 22092. Annual ring widths of Quercus a I ba and L Ir lodendron tulip! fera were used to analyze monthly response of radial tree growth to temperature and precipitation. Each species was sampled from two meslc sites: on soil underlain by limestone conglomerate, and on soil derived from metamorphosed schist. Monthly responses to climate were derived by multiple regression of standardized tree ring widths with principal components of climate. LI r lodendron from the limestone site seems more responsive to temperature than to precipitation. Radial growth was correlated with low temperatures In May and July, and with high precipitation In July. Thus, for both sites, Quercus a I ba and L I r I odendron may be most valuable for reconstruction of late spring and early summer climate. LICHEN SECONDARY COMPOUND INHIBITION OF LICHEN ASC0SP0RE GERMINATION. John C. Whiton* and James D. Lawrey, Dept, of Biol., George Mason Univ., Fairfax, V a. 22030. Germination responses of ascospores of three lichenized fungi, Cladonia cristatella , Graphis scripta and Caloplaca citrinia, and the nonl ichenized ascomycete Sordaria fimicola, were observed in the presence of pure lichen acids. Acetone solutions of lichen acid at a concentration of 2.7 X 10"3 M were poured onto buffered agar ranging in pH from 4 to 7 , the acetone was eva¬ porated and spores deposited on the agar. Treatment plates received either evernic or stictic or vulpinic acid or atranorin. Control spores were not ex¬ posed to lichen acid. Evernic and vulpinic acid usually reduced or completely inhibited germination. Stictic acid and atranorin were usually nontoxic. Lichen acid toxicity was independent of pH. There was no apparent differen¬ tial roxicity toward the nonl ichenized Sordaria or toward crustose species. Lichen acids have allelopathic potential, thus, they may affect the structure and development of natural plant communities. RESPONSES OF TERRESTRIAL MOSSES TO S02 AND DESICCATION. William E. Winner, Dept, of Plant Pathol. & Physiol., Va. Polytechnic Inst. & State Univ., Blacks¬ burg, VA 2 406 1 . Gas exchange experiments were used to define the photosynthe¬ tic responses of 3 common feather moss species to both desiccation and SO2. P 1 euroz i urn schreber i , Hylocomi urn splendens , and Pt i I i urn cr i sta-castrens i s were studied. Experiments included simultaneous measurements of desiccation, net photosynthesis, and SO2 uptake. P_. schreber i seemed the most sensitive species to desiccation and SO2, both alone and in combination. The immediate effects of SO2 upon photosynthesis of fully hydrated and partially dried mosses were simi¬ lar. Only mosses fumigated in the fully hydrated condition retained photosyn¬ thetic impairment following a 24 h recovery period. This probably is related to the larger amounts of SO2 absorbed by fully hydrated plants. Thus SO2 resis¬ tance of mosses changes as they desiccate. CHANGES IN THE VESSEL ANATOMY OF FLOOD-DAMAGED TREES. Thomas Ik Yanosky*, U. S. Geol. Survey, Reston, Va. 22092. Concentric bands of abnormally large vessels were detected in the latewood of ash trees (Fraxi nus americana L. and F. pennsy I van ica Marsh.) sampled along the Potomac River near Washington, D.C. Tangential diameters of vessels ranged from 60-160 >*m, compared to 30-60 for typical latewood vessels. Tyloses were generally apparent. Within a particular annual ring, the largest and most closely spaced vessels formed near the apex, and became smaller and diffuse basipetally. In some trees, vessels developed only near apices. It is hypothesized that abnormal vessels develop when flood-stripped crowns produce a second crop of leaves. Supporting evidence is that vessels formed only during years of known flooding, and only if trees had been totally submerged. Additionally, trees experimentally defoliated formed similar growth patterns upon refoliation. Variations in the size and spacing of vessels at different stem heights suggest that development is controlled in large part by the polar movement of growth regulators along a diminishing gradient from expanding buds and leaves. THE VIRGINIA JOURNAL OF SCIENCE 145 EVIDENCE OF HIGH STREAMFLOW LEVELS FROM THE LATE WOOD FIBERS OF FLOOD-PLAIN TREES. Thomas M. Yanosky , U. S. Geo I . Survey, Reston, Va. 22092. Changes in the transverse dimensions of latewood fibers were studied in growth rings of ash trees (Fraxinus americana L. and £. pennsy I van ica Marsh.) collected along the Potomac River near Washington, D. C. Concentric bands of fibers with atypical ly large lumens and thin walls were commonly detected in trees growing near the low-water channel. Streamf I ow records indicate that roots of most trees with these "white rings" were flooded during the I atewood-grow th interval. Trees apparently were not damaged and a concomitant reduction in water stress seems to have accelerated the rate of radial growth. The intra¬ annual position of anomalous fibers general ly corresponded to the time of increased streamf low within the estimated interval of latewood growth. For example, white rings in the outer latewood coincided with high flows in late July or August. Trees occasionally formed anomalous fibers even though roots were not flooded, although associated discharges were significantly greater than the corresponding value of the 50-year monthly average. These studies may provide insights into the rate and timing of radial growth and might have applications for streamf I ow-reconstruct ion techniques. A COMPARISON OF CHLOROPHYLL A EXTRACTION EFFICIENCY FOR PHYTOPLANKTON. P. V. Zimba* , W. Dunstan, and H. G. Marshall. Departments of Biological Sciences and Oceanography, Old Dominion University, Norfolk, Va. 23508. Chlorophyll a_ concentration was determined using two extractants - DMSO : Acetone (4:6 v/v) and aqueous acetone (90%) . The method of Burnison (1980) was modified, eliminating the necessity of two f iltrations . A Turner Designs f luorometer was used for the chlorophyll measurement on six phytoplankton cultures that included both fresh water and marine species . The cultures con¬ sisted of a cyanophycean, three chlorophyceans , a diatom, and a diatom mixture. In all cases the DMSO: Acetone extraction procedure was at least as efficient as the use of aqueous acetone . When DMSO : Acetone was used to extract Skeletonema costatum and two chlorophyceans (Volvox aureus and Carteria sp. ) , higher chlorophyll a_ values were obtained (p=0.05) . The use of the DMSO : Acetone extraction on marine cultures has not been reported previously in the literature . COMPARISON OF LEAF CHLOROPLAST ULTRASTRUCTURE IN C3 and C4 MONOCOTS . P. V. Zimba* , V. Wang* , and K. K. Nesius . Department of Biological Sciences, Old Dominion University, Norfolk, Va. 23508 . Avena sativa (C3) and Zea mays (C4) chloroplast thin sectio ns were examined by transmission electron microscopy for proportion of constituent stroma and lamellae . Avena mesophyll chloroplasts consisted of 65% lamella and 35% stroma . Zea mesophyll chloroplasts located 7 cells towards the leaf margin consisted of 86% lamella and 14% stroma ; whereas, mesophyll chloroplasts adjacent to the bundle sheath consisted of 78% lamella and 22% stroma . The bundle sheath chloroplasts contained 13% lamella and 87% stroma . Chemistry Section THE DEPENDENCE OF LEACHING OF CATIONS FROM SEDIMENTARY ROCKS ON pH; AN ACID RAIN ANALYSIS . Daniel Armstrong, Robert T. Grauer Jr. , David S. Knitter , Michael A. Pleva, Scott S. Prysi, James K. Shiilington, Richard W. Young, Dept . of Chem. , Washington and Lee Univ. , Lexington, VA 24450 . For the purposes of our research both natural and synthetic acid rain. Samples of various sed imentary rocks of known mass were introduced into a predetermined volume of the acid rain. Both the cation leaching (measured in terms of dissolved solids) and the pH variation were measured over time. Current data suggests that the amount of cation leaching is determined by the pH of the acid rain medium and on the type of rock introduced. 146 THE VIRGINIA JOURNAL OF SCIENCE INVESTIGATION OF SOLVENT PAIR RATIOS DURING PROGRESSION ALONG REVERSED-PHASE THIN LAYER CHROMATOGRAPHIC PLATES FOR THE PURPOSE OF FRACTIONATING HOMOPOLYMERS Daniel W. Armstrong, D_^ James Bailey III , John W. Miller III, James K. Shillington, Robert B. Tucker, Jr., Dept, of Chem. , Washington and Lee Univ. , Lexington, VA 24450. The composition change of the mobile phase along the length of the chromatogram allows one to fractionate polystyrene homopolymers by reversed-phase thin layer chromatography. The adsorption of the more hydrophobic component of the solvent pair to the hydrophobic stationary phase causes this gradient change. If the composition change can be calculated as the solvent pair travels along the chromatogram, the best solvent ratio for the fractionation of polymers of particular molecular weights can be found. THE STRUCTURAL HYPOTHESIS OF CANCER: A ROLE FOR THE CELL MEMBRANE IN BIO¬ CHEMICAL EQUILIBRIUM. I\_ GL_ Barber , Department of Natural Sciences, Long- wood College, Farmville, Virginia 23901. A hypothesis is presented and current literature cited to suggest that cell membranes have a major role in the regulation of cell function and the maintenance of cellular equilibrium, which complements the role of the genetic material. Small, active molecules such as superoxide ion have an important function in this regulation by modi¬ fying membrane behavior. The production of proteins completes the feed-back loop by providing a further means of communication between the cell wall, the cell nucleus, and other organelles. 2-ALKYLIDENE-l , 3-CYCL0PENTANEDI0NES : VERSATILE SYNTHETIC INTERMEDIATES. William H. Bunnelle*, Dept, of Chem., Col. of William and Mary, Williamsburg, Va . 23185. Sulfenylat ion-oxidative elimination of 2-alkyl-l , 3-cyclopentane- diones provides a mild and simple route to 2-alkyl idene-1 , 3-cyclopentanediones a heretofore elusive class of compounds. Their high reactivity towards carbon carbon bond-forming reactions, notably Diels-Alder additions, coupled with the synthetic flexibility of the cyclopen tanedione moiety fortell a promise of extraordinary synthetic utility for these compounds. The utility of 2-phenyl- thio-1 , 3-cyclopen tanedione as a synthon for this system further extends the adaptability of this method. STUDIES ON EXTRACTABLE CONSTITUENTS OF FOMES ROBINIAE. Peyton Frost Burnett* and James B. Patrick, John Baker Daffin Dept, of Chemistry, Mary Baldwin Coll. Staunton, Va. 24401. Fomes robiniae (Murr.) Sacc. et D. Sacc. ( Polyp oraceae) is a woody bracket fungus occurring abundantly in the central Shenandoah Valley. Extraction of the dried and pulverized fungus with several solvents has yielded a variety of, products after pH fractionation of the extracts. The acetone extract yields a significant quantity of a neutral fraction composed predominantly of two components separable by flash chromatography. We report herein the results of preliminary characterization of these two components by nuclear magnetic resonance, infrared, and ultraviolet spectro¬ metry and wet chemical tests. POLYMERIC ANTIBIOTICS OF GRAMICIDIN-S. Susan L^_ Carstens and Patrick G. Barber, Department of Natural Sciences, Longwood College, Farmville, Virginia 23901. Gramicidin-S is a cyclic decapeptide antibiotic that contains two amine groups on the ornithine amino acid residues. These were used to prepare a series of polymers by condensation with several diacyl chlorides of in¬ creasing chain length. The resulting polymers retained antibiotic behavior against gram positive bacteria, and this retention was in proportion to the amount of gramicidin-S incorporated into the polymer. These results are in conflict with those reported in the literature which indicate that the anti¬ biotic activity of gramicidin-S is lost upon acylating the ornithine amine groups. THE VIRGINIA JOURNAL OF SCIENCE 147 pH-INDUCED CONFORMATIONAL TRANSITION OF P0LY( ITACONIC ACID-CO-STYRENE). C. Y. Chung and R. M. Ottenbrite, Dept, of Chem., Virginia Commonwealth Univ., Rich¬ mond, Va. 23284. A highly regular structure of itaconic acid-styrene copoly¬ mer was prepared by free radical polymerization and separated into two molecu¬ lar weight fractions 1,000-10,000 and 10,000-30,000. This copolymer was char¬ acterized by FT-IR, elemental analysis and FT-C13 NMR. These analysis indicate that the polymer has an alternating head-to-tail structure in both molecular weight fractions. The effects of counterion binding and ionic strength of lithium, sodium and TMA ions on the pH-induced conformational transition was evaluated by potentiometric titrations and viscometric titrations. The order or ionic binding was found to be Li+>Na+>TMA+ in both aqueous and salt solu¬ tions. The effect of ionic strength of the counterion (salt effect) has a predominent influence in the case of lithium and sodium ions, but only a small role in the case of TMA ions. The free energy change (AG°) was evaluated and correlated with the conformational change for each transition. The low mole¬ cular weight fraction appears to be more sensitive to site binding on the poly¬ mer and to ionic strength of the solution than the higher molecular weight frac¬ tion. Viscosity data indicates that molecular expansion during the titration process for the high MW fraction is much greater than in the low MW fraction. AN EASY, GENERAL SYNTHESIS OF ARYL SULFONES. Donald W. Clary,* Herbert J. Sipe, Jr., and Samuel B. White,* Department of Chemistry, Hampden-Sydney College, Hampden-Sydney, VA 23943. An improved synthesis of aryl sulfones is reported. This improved synthesis, a modification of that reported by Graybill [£f. J_. Org. Chem. , 1967 , 32 , 29311 , consists of adding phosphorous pentoxide to the polyphosphoric acid reaction medium and to the appropriate sulfonic acid reagent. Syntheses of phenyl-tolyl sulfone, diphenyl sulfone, biphenyl-phenyl sulfone, and ditolyl sulfone by this modified procedure resulted in substantial increases in yield over those reported by Graybill. SELECTIVITY ENHANCEMENT STUDIES OF POTENTIOMETRIC BACTERIAL ELECTRODES. Carolyn A. Corcoran* and Robert K. Kobos , Dept, of Chem., Va. Commonwealth Univ., Rich¬ mond, Va. 23284. The selectivity of a previously reported bacterial arginine electrode has been greatly enhanced, with no loss of response to L-arginine. The arginine electrode is a potentiometric sensor which consists of intact bacterial cells (Streptococcus faecium) coupled with an ammonia gas-sensing electrode. This electrode has been shown to exhibit a near-Nernst i an response to L-arginine in the concentration range of 4.0 x 10”^ M to 1.5 x 1 0“ 3 m. However, glutamine is a serious interference if it is present in the sample solution, thereby lim¬ iting the application of the sensor in biological fluids. In this study, it was found that the response to glutamine was negligible initially but increased daily, possibly due to the presence of a contaminant in the bacterial layer of the electrode. It was shown that using 1.0 x 10“3 M sodium azide in the working buffer and storing the electrode at 0°C effectively maintains the initial low response to glutamine in the concentration range of 4.0 x 10“5 M to 1.5 x 10“3 M, without reducing the response to L-arginine. When this procedure is followed, the sensor can be used to measure L-arginine in the presence of up to 1 x 1 0_ 3 M glutamine. AN APPLICATION OF THE BENEDETTI -PICKLER EQUATION IN THE SAMPLING OF COMPLEX MIXTURES FOR CHEMICAL ANALYSIS. John J. Delany, III, M. A. Pleva, Dept, of Chem., Washington and Lee Univ., Lexington, VA 24450. An area of analytical chemistry of considerable importance is sampling and sampling errors. Sampling is usually not considered in analytical chemistry courses because of its complexity. This work demonstrates the problems of sampling and their solution using the Benedetti-Pickler and hopefully facilitates the understand¬ ing of the sampling process. 148 THE VIRGINIA JOURNAL OF SCIENCE MECHANISM FOR THE AQUEOUS DECOMPOSITION OF 1-PIPERIDINOCYCLOHEXANE CARBONITRILE. M. P. DiFazio* and W. H. Soine, Dept, of Pharm. Chem. , MCV/VCU, Richmond, VA 23298. The presence of piperidinocyclohexanecarbonitrile (PCC) in illicit phen¬ cyclidine is of toxicological importance because it decomposes to release cya¬ nide. This study was directed toward determining the mechanism by which cyanide was released. PCC was observed to undergo pseudo first order decomposition in buffered solution and the rate of decomposition was found to be dependent on the pH of the buffer. Initially, the rate of decomposition increased from pH 2 to 5.6 where a maxima was reached. The rate then decreased until it reached a min¬ ima at pH 8.3 followed by a second increase in rate of decomposition to pH 13. These results indicate there is both an acid catalyzed and base catalyzed mecha¬ nism for the release of cyanide and suggests that PCC decomposes through both an imine and enamine intermediate (Supported by the Grants-in-Aid Program for Faculty at Virginia Commonwealth University). THE EFFECTS OF TRANS FATTY ACIDS ON A5 FATTY ACYL DESATURASE ACTIVITY IN HUMAN SKIN FIBROBLASTS. Mark A. Doloresco* , Dept, of Chem., Old Dominion Univ., Norfolk, VA 23508, and Miriam D. Rosenthal, Dept, of Biochem. , Eastern Va. Med. School, Norfolk, VA 23501. Fatty acyl A5 desaturase converts eicosatrienoate (20:3) to arachidonate. Dietary trans fatty acids alter essential fatty acid metabolism in vivo. We investigated the effects of trans fatty acids on A5 desaturation. Human skin fibroblasts were preincubated for 2h with trans-v accenate (lit 18:1), elaidate (9t 18:1) or linoelaidate (9t 12t 1 8 : 2 ) . 2. 5yM[ 1 4C] Eicosatrienoate was then added. After 6h the cells were harvested and their lipids extracted. [14C] Fatty acid methyl esters were prepared, and analyzed by radio-gas chromato¬ graphy. Control cells desaturated 58? of the incorporated [14C]20:3. Desatura¬ tion was 20?, 33? » and 46? for cells given 40pM elaidate, linoelaidate and trans- vaccenate respectively. Elaidate was as inhibitory as arachidonate. Thus inhibition by trans 18:1 isomers is influenced by the position of the double bond. (Supported in part by NIH Grant HL-23658.) SYNTHESIS OF 5-3H-l,2,3-BENZENETRIOL (PYROGALLOL) . D. M. Dulik* and W. H. Soine, Dept, of Pharm. Chem., MCV/VCU, Richmond, VA 23298. In order to facilitate me¬ tabolic studies of pyrogallol (1,2,3-benzenetriol, PY) , it was necessary to ra¬ diolabel PY with tritium in a chemically stable position. Model reactions using deuterium resulted in the formation of 4,6-2H2“PY under both acid and base cata¬ lyzed conditions. However, under simulated physiological conditions, it was ob¬ served that the label underwent facile exchange. Alternatively, 5-2H-PY was prepared via a two step synthesis. Transmetallation of 3,4,5-trimethoxybromo- benzene (I) with n-butyllithium in dry hexanes at -70 C to -30 C followed by quenching with excess D2O gave 5-2H-l , 2 , 3-trimethoxybenzene (II). 0-demethyla- tion of (II) with BBr3 and HBr in CH3COOH gave 5-2H-PY (III) in 30% overall yield. 5-2H-PY was found to be stable to exchange under physiological condi¬ tions. This method was employed to prepare 5-^H-PY with specific activity of 7.6 mCi/mmole and of suitable purity for use in in vivo metabolism studies. CHEMISTRY AND THE CONSERVATION OF ARTIFACTS IN THE SOVIET UNION. Vera B. Espinola, Ethnographic Objects Conservator, 3^34 Camelot Drive, Annandale , Va. 22003. A report is given of research and analytical chemistry as it is utilized in the service of conser¬ vation at the All Union Scientific and Research Institute for Restoration (VNIIR) in Moscow. Solvents and adhesives used in the major restoration centers of Moscow and Leningrad are fairly standardized with a preference for natural organic adhesives in the conservation of icons and textiles. Synthetic polymers are approached cautiously and observed in accelerated aging tests to determine if they conform to the principle of reversibility. The use of ftorlon and ftoroplast , a recently developed polytetra- flouroethylene product is discussed. A brief account is given of health hazards in the handling of some conservation materials. (IREX grant, USSR.1980) THE VIRGINIA JOURNAL OF SCIENCE 149 PREPARATION AND STUDY OF ELECTRICAL CONDUCTIVITY OF A HETERO AROMATIC LADDER POLYMER: BBL. Patricia Etienne* and George W. Mushrush, George Mason Univ., Fairfax, VA 22030 and David C. Weber,* Naval Research Laboratory, Washington, DC 20375. Synthetic organic metals have recently been the subject of extensive research efforts. Polyacetylene, (CH) , is the simplest conjugated polymer with a planar backbone. Doping enhances the electrical conductivity of (CH) by twelve (12) order of magnitude. Graphite, also a planar material, exhibits intrinsic metal-like conductivity that can be increased by doping (inter¬ calation) . Ladder polymers such as benzimidazobenzophenanthroline (BBL) have struc¬ turally rigid, planar geometry. Because BBL incorporates the conjugated planar backbone structure of polyacetylene with an extended ring network similar to graphite, we are presently studying the effects of chemical doping on the elec¬ trical conductivity of BBL. The BBL ladder polymer was synthesized by condensation polymerization of 1 ,4,5,8-Naphthalenetetracarboxylic acid (NTCA) and 1 , 2 , 4 , 5-Tetraaminobenzene tetrahydrochloride (TAB-4HC1). Films were prepared by precipitation from methanesulfonic acid and vacuum filtration on a glass frit. Chemical doping procedures and electrical conductivity measurements will be discussed. A HALF-CENTURY OF CHEMISTRY : PERSONAL EXPERIENCES AND VIEWPOINTS. Hyman I. Felna teln*, 10411 Forest Ave., Fairfax, VA 22030. The story of one chemist starting his career at the heglnnlng of the great depression will be traced through a teaching apprentice¬ ship, government service, and back to teaching before retirement. Chemists In academe and government who exerted an Important in¬ fluence on this career, practical problems and the thought proc¬ esses that helped solve them, research and publications, peer and referee review, professional and subprofessional teaching, the role of descriptive and theoretical chemistry, and serendipity, will all be frankly discussed as time will allow. RECENT SYNTHETIC ADVANCES IN THE ORGANOMETALLIC CHEMISTRY OF CHLOROCYCLOTRI- PHOSPHAZENES . Paul J. Harris and Logan A. Jackson*, Dept, of Chemistry, V a. Polytechnic Inst, and State Univ., Blacksburg, V a. 24061. The organometal 1 ic chemistry of cyclic chlorophosphazenes has been a sub¬ ject of intense study in recent years. These reactions are found to proceed via a variety of different pathways including metal -halogen exchange and ring cleavage. Only recently have reactions that proceed via substitution of halogen been observed. The reactions of organotitanium reagents are found to yield monoal kylated phosphazenes whereas organoal uminum reagents are found to yield a variety of highly alkylated phosphazene compounds. The mechanistic aspects of these reactions will be discussed along with the spectroscopic characterization of the products. INVESTIGATION OF THE EFFECT OF A SHORTENED INCUBATION PERIOD ON THE RELIABILITY OF THE RADIOIMMUNOASSAY FOR CORTICOTROPIN. Anne R. Keller*, Dept, of Biochem. , McGill Univ., Montreal, Quebec, Canada. The Radioimmunoassay (RIA) method originally developed by Berson and Yalow for determining corticotropin (ACTH) levels employs a six day incubation in order to achieve maximum sensitivity. Workers using shorter incubations have not studied the extent of change in sensiti¬ vity. This paper compares the sensitivity of the ACTH assay for standard curves run for 1/4 to 2 hours at 25° C with curves run for 3-7 days at 4° C. Precision was shown to be equal in the two methods and sensitivity was more than adequate for the analysis of the hormone present in cultured cell preparations which were the subject of our investigations. It was concluded that the use of a shortened incubation period is valid and that the advantages in time saved more than offset the decrease in sensitivity observed. 150 THE VIRGINIA JOURNAL OF SCIENCE REACTIONS OF Mo(VI) SCHIFF BASE COMPLEXES WITH ETHYLDIPHENYLPHOSPHINE . James T. Lyon, III and Joseph Topich, Department of Chemistry, Virginia Commonwealth University, Richmond, VA 23284. The reaction of ethyldiphenylphosphine with a number of ci s-dioxomolybdenum(VI ) coordination complexes are described. These complexes incorporate tri dentate Schiff base ligands obtained from the conden¬ sation of 5-X-sal i cyl al dehyde (X = Cl , Br, H, CH30) with o-aminobenzenethiol or 2-aminoethanethiol . Oxo-molybdenum( IV) Schiff base complexes were observed as products of the reaction of these Mo(VI) complexes with PEt02. The kinetics for these reactions were followed spectrophotometrical ly and the applicable rate law i s-d[Mo02L]/dt = ki[MoO2L][PEt02] . The ki's are shown to vary system¬ atically as the X-substituent on the ligand is changed. It was also observed that a correlation exists between the cathodic reduction potentials (Epc) and the ki's within each series. There are several ligand variations whose effects systematically alter the Mo(VI) ki's and Epc 1 s . These individual effects will be considered separately and their cumulative effect will also be described. STUDIES ON EXTRACTABLE CONSTITUENTS OF POLYPORUS SULFUREUS. Michelle D. Makanas* and James B. Patrick, John Baker Daffin Department of Chemistry, Mary Baldwin College, Staunton, Va. 24401. A large specimen of the fleshy fungus Polyporus sulfureus was subjected to successive Soxhlet extractions with several different organic solvents. Each extract was subfractionated in turn into strong and weak acidic, basic, and neutral fractions. In this way several significant fractions were obtained which were further separated and purified by flash chromatography and Kugelrohr distillation. None of the materials so obtained has yet been identified, but we report preliminary chemical and spectrometric characterization data and details of the isolations. CARBON-13 NUCLEAR MAGNETIC RESONANCE SPECTRA OF SOME MES0I0NIC XANTHINE ANALOGS. G.O. Mbagwu , R.G. Bass and R.A. Glennon, Depts. of Chem. and Pharm. Chem. , Vir¬ ginia Commonwealth Univ., Richmond, Va. 23284. Natural abundance 1 3c NMR chemical shifts have been experimentally determined for a series of mesoionic th i azolo[3 ,2-a] py r i mi d i ne-5 , 7”d i ones . The spectral data are compared to those of related mesoionic d i hydroth i azolo[3 ,2-a] pyrimi¬ dine-5 , 7-d i ones and mesoionic 1 , 3 , 4- th i ad i azol o[3 ,2-a] py r imi d i ne~5 , 7~d iones . Reasonable correlation between the observed 1 3c NMR chemical shifts and CNDO/ 2 total charge densities have been obtained for the different carbon atoms of 8- methyl th i azol o[3 ,2-a] py r i mi d i ne~5 , 7-d i one . The support of this research in part by U.S. Public Health Services (Grant No. HL-22566) and the School of Graduate Studies, Virginia Commonwealth Univer¬ sity is gratefully acknowledged. AN INDUSTRIAL SABBATICAL. William H. Myers, Dept, of Chem., Univ. of Richmond, Richmond, Va. 23173. The author spent a fifteen month period from May of 1981 to August of 1982 on sabbatical leave, working in the Research and Development Laboratories of Ethyl Corporation in Baton Rouge, Louisiana. A brief report on the experience will be given, along with suggestions to those who might consider doing something similar. The suggestions will include both observations on the benefits likely to accrue to the academic chemist and to the industrial research group and cautions to consider to make the experience go more smoothly. THE VIRGINIA JOURNAL OF SCIENCE 151 AROMATIC AMINE - HALOBORANES : EVIDENCE FOR TRIGONAL BORONIUM CATIONS. W. H. Myers and D. H. Reid* Dept, of Chem. , Univ. of Richmond, Richmond, Va. 23173. The halogenation reactions of borane adducts of 2-picoline, 2 , 6-lutadine , 3,5- lutadine, and 2,4,6-collidine were monitored by 'H nmr. Signals due to both aliphatic and aromatic protons in the various adducts showed a nownfield shift as size and/or number of halpgens increased. These downfield shifts can be accounted for by a combination of steric compression and inductive effects. The last step of halogenation is accompanied by an upfield shift of the signals due to protons in the 2 or 6 positions of the pyridine ring, a further down- field shift of signals due to protons in the 3, 4, or 5 positions of the ring, as well as by the appearance of conductivity in the solution. These observa¬ tions are attributed to the formation in solution of trigonal boronium cations. These compounds have been isolated and characterized. CALORIMETRIC STUDIES OF ACTIVE-SITE LIGAND INTERACTIONS WITH GLUTAMINE SYNTHETASE FROM E. COLI. Sue H. Neece,* Eileen G. Gorman,* & Ann Ginsburg,* NHLBI , NIH, Bethesda , MD 20205. Heats of binding the active-site ligands L-glu, the L-glu analog L-met-S-sulf oximine , and the ATP analog adenylyl- imidodiphosphate (AMP-PNP) to the unadenylylated , manganese form of dodecameric glutamine synthetase (GS) from E_. coli were measured in a batch-type calorimeter at pH 7.1 and 30°C. Separate determinations in buffers with different heats of protonation indicated that proton effects were negligible for each binding reaction. Values of AH (± 1 kcal/mol) are given per mole of enzyme subunit for saturation with ligand: For L-glu, AH = -6.2; for L-met-S-sulf oximine , AH = -15.1; for AMP-PNP, AH = +1.3; for L-met-S-sulf oximine binding to the Mn • G S • AMP-PNP complex, AH = -13.5 kcal/mol. The binding of L-glutamate is considerably less exothermic than that of the L-glutamate analog, and in contrast to AH = -5.7 kcal/mol for ADP binding to Mn*GS (Shrake et al. , Biochemistry 16, 4372, 1977), the binding of AMP-PNP is slightly endothermic. TI TRIMETRIC AND ULTRAVIOLET STUDIES ON ALKYL HYPOBROMITE. Nina M. Roscher and Chinh K. Nguyen* Dept, of Chemistry, The American Univ., Washington, DC 20016 Alkyl hypobromites are prepared by treating tertiary alcohols with bromine using mercuric oxide as the catalyst with pentane as the solvent. The forma¬ tion of the alkyl hypobromites was confirmed by ultraviolet and titrimetric data in this study. Spectropho tometric measurements of samples of the reaction mixture were made on a DMS 90 UV-Visible spectrophotometer. Ultraviolet scans were performed from 440 to 240 nm to confirm the disappearance of bromine at 415 nm and the presence of the alkyl hypobromite at 280 nm. In the titrimetric studies, samples of the reaction mixture were quenched by an acidified potassium iodide solution. The resulting mixture was titrated against a sodium thiosulfate solution and then against a sodium hydroxide solution. The titrimetric results indicated an appreciable yield of the alkyl hypobromite. 152 THE VIRGINIA JOURNAL OF SCIENCE FURTHER STUDIES ON THE ISOLATION AND CHARACTERIZATION OF VIOLACEIN. Jo Ann Noble’' and Nina M. Roscher’', Dept, of Chemistry, The American Univ., Washington, D. C. 20016. Based on the results of synthetic studies by earlier researchers, the following structure has been proposed for violacein: In an attempt to confirm the above structure via modern spectroscopic tech¬ niques including UV, IR, NMR and mass spectroscopy, the properties of violacein in various organic solvents was examined. Further characterization of violacein was investigated by performing the standard test for siderophores , Iron-chelat¬ ing substances. The results from these preliminary studies will be presented. THE IDENTIFICATION OF PRODUCTS FORMED IN TgE EP0XIDATI0N OF M0N0- TERT-BUTfL-p-BENZUQUINQNE. Eahnie M, PharrT'and Elizabeth M. Hair- field, John baker Daffin Laboratory, Department of Chemistry, Mary Baldwin College, Staunton, VA 2I4I4.OI . Many workers have studied the importance of epoxides in metabolism. In addition, epoxy groups are present in numerous antitumor antibiotics and carcinogens. In this research project, the epoxidation reaction of mono- ter t-butyl- £-benzoquinone with alkaline hydrogen peroxide is investigated. Extraction methods, flash chromatography, thin-layer chromatography and spectroscopic studies are carried out to classify the products formed from the epoxidation reaction with alkaline hydrogen pero¬ xide. Results obtained show the presence of four products formed, one of which is an epoxide, and the remaining three products are thought to be carboxylic acids. Further work is required to con¬ firm the presence of carboxylic acids. The difficulties encoun¬ tered in the study are discussed. MUTUAL INTERACTIONS OF IRON AND URANIUM REDOX STATES IN GLASS MELTS USED FOR NUCLEAR WASTE IMMOBILIZATION. Henry D. Schreiber, Department of Chemistry, Virginia Military Institute, Lexington, VA 24450; Leslie M. Minnix & Barbara E. Carpenter, College of William and Mary; G. Bryan Balazs, Washington and Lee University; and Todd N. Solberg, Virginia Polytechnic Institute. Uranium establishes the U ( V I )-U (V )-U ( I V ) redox equilibria in borosilicate glass melts, while iron can exist as Fe (III), Fe(II), or Fe (0 ) in equivalent melts. Borosilicate glasses used in nuclear waste immobilization are required to incorporate uranium into their structures in the presence of the multivalent element iron. A basic understanding of the mutual interactions by electron exchange of the uranium and iron redox states will aid in the scientific basis for nuclear waste immobilization. For one-electron reductions in borosilicate melts at 1150°C at a particular oxygen fugacity, U ( VI ) is easier to reduce than is Fe(III), which in turn is easier than U(V). Fe(III) does not oxidize uranium, nor does Fe(II) reduce uranium. THE VIRGINIA JOURNAL OF SCIENCE 153 A NEW SYNTHESIS OF 4-HALO-5 , 6 -DIHYDRO- 2H-PYRANS . D. W. Seamon*, W. H. Bunnelle, and D. W. Thompson, Dept, of Chem., Col. of William and Mary, Williamsburg, VA 23185. The acetals derived from homopropargyl alcohols and ethyl vinyl ether undergo Lewis acid catalyzed cyclization to give 4 -halo-5, 6-dIhydro-2H-pyrans (eqn 1). fH3 TiCl ch3ch2ochoch2ch2cech ch c-3 > 90% Results with several acetals and Lewis acids will be presented. UTILIZATION OF THE HEWLETT PACKARD 5970 GC/MS FOR THE ROUTINE CONFIRMATION OF THE 1 1-NOR- DELTA-9 -CARBOXYLIC ACID URINARY METABOLITE OF MARIHUANA. Robert K. Simon and R. Hugh Granger, AMERICAN MED. LABS., INC. Rapid, low cost confirmation of the urinary metabolite of marihuana, ll-nor-delta-9-COOH is achieved with the new HP capillary GC/MS system which features air cooled pumps and simple, convenient attachment to many capillary GC units. Deuterated 1 1-nor-del ta-9-COOH serves as the internal standard for the method. Hydrolyzed urine is extracted with hexane/ethyl acetate and the metabolite is derivatized with both TMAH in DMSO and I ODOMETH ANE to yield the methyl ether, methyl ester derivative. Comparison of this procedure to standard packed column El and Cl methods show that the 5970 procedure is selctive, sensitive and precise. A detection limit of 1 nanogram per milliliter has been documented. Correlation of the described procedure with the Syva Emit DAU marihuana urine screen shows greater than 95% correlation in specimens providing an immunological response greater than 50 ng/ml . The relative low cost of the HP 5970 makes it attractive as a GC/MS alternative for labs conducting drug testing. AN ELONGATED FORM OF 5.8S rRNA: A GENE VARIATION. Dave Smith and Thomas 0. Sitz, Department of Biochemistry and Nutrition, Virginia Tech, Blacksburg, VA 24061. The 5.8S rRNA is found in eucaryotic cells bound to the 28S rRNA in the large ribosomal subunit. When whole cell RNA preparations isolated from rat or mouse tissue were separated on high resolution polyacryla¬ mide gels, two bands of 5.8S rRNA were observed, a major band followed by a less intense minor band. This minor form represented 15 to 40% of the total 5.8S rRNA depending on the source of tissue. Sequence analysis and the kinetics of formation showed that this minor form was elongated at the 5' end and is not a precursor. The sequence of the minor form was found to be p(C)CGAUACG--which is five or six nucleotides longer than the major form and has a heterogenous termini. The DNA sequence for this region in rat has been reported to be -- CCGTACG This would suggest that gene heterogeneity leads to an insertion of an A residue in the ribosomal precursor RNA which changes the processing site. 154 THE VIRGINIA JOURNAL OF SCIENCE SYNTHESIS OF 2 SUBSTITUTED 4 , 6-DIHYDROXYPYRIMIDINES . F. T. Smith*, W. H. Soine and B. van't Riet, Dept, of Pharm. Chem., MCV/VCU, Richmond, VA 23298. Inhibi¬ tors of ribonucleotide reductase have been shown to be active against L1210 leu¬ kemia. Several compounds containing the catechol function with a planar substi¬ tuent are known to be effective against this enzyme. In order to investigate the importance of the catechol functionality we have prepared a number of 2 sub¬ stituted 4,6-dihydroxy pyrimidines. The appropriate acid was converted to the acid chloride and subsequently to the amide. Thionyl chloride was used to dehy¬ drate the amide to the corresponding nitrile. The Pinner synthesis gave the imino ester which upon reaction with ammonia gave the corresponding amidines. The amidines could be easily condensed with diethyl malonate in sodium ethoxide to give the corresponding 2 substituted 4,6-dihydroxy pyrimidines in good yields. The 2 substituent included methyl, phenyl, 3-hydroxy-, 4-hydroxy and 3,4-dihy- droxyphenyl. (Supported by the Jeffress Memorial Trust). THERMAL REDUCTION OF CHLORDECONE IN THE PRESENCE OF ALCOHOL. W. H. Soine, T. R. Forrest* and J. D. Smith, Dept, of Pharm. Chem., MCV/VCU, Richmond, VA 23298. It was observed that chlordecone (CD) underwent reduction to chlordecone alcohol (CDOH) if a methanol solution of CD was analyzed using gas chromato¬ graphy. This study was to determine the conditions necessary for this reaction and steps for minimizing or preventing the reduction. Both primary and secon¬ dary alcohols when co-injected with CD were found to cause the reduction while no reduction was observed with ^-butanol or aprotic solvents. In general, the hotter injection port (240 to 360°C) caused more reduction to occur. When CD was heated with benzyl alcohol the major products formed were CDOH and benzalde- hyde. Finally, simultaneous injection of CD, alcohol and magnesium chloride in¬ creased reduction of CD relative to a mixture without magnesium. Based on these results it appears that the reduction was thermally catalyzed in the injection port and appears to go via a mechanism comparable to the Meerwein-Ponndorf-Verley reduction. SYNTHESIS AND BIOLOGICAL EVALUATION OF 103> 106ru-B-RUTHENOCENYLALANINE. W. H. Soine , Dept, of Pharm. Chem., MCV/VCU, Richmond, VA 23298; C. E. Guyer* and F. F. Knapp, Jr.*, Nuclear Medicine Group, Oak Ridge National Laboratory, Oak Ridge, TN. The potential application of the ruthenium radionuclides in diagnos¬ tic medicine necessitated the synthesis of radiolabeled ruthenocenes in which the side chain could be modified to provide tissue specificity. This required synthesis of the synthetically versatile quaternary ammonium salt, 103,106ru_ ruthenocenylmethyltrimethyl ammonium iodide (I). Carrier free 103, 106Ru_ruthe- nocene was synthesized from 103,106ruci3 using TiCl3«H2 and cyclopentadienylso- dium. The ruthenocene ring was functionalized using the Mannich reaction to give 103, lObRu-dimethylaminomethylruthenocene which was quaternized with methyl- iodide to give I in 20% overall yield. From this intermediate we prepared car¬ rier free 103 , 106Ru_g_ruthenoceny;Laianine (n) by homologation of the side chain using sodio diethylf ormomidomalonate followed by basic hydrolysis to II. II was evaluated as a pancreatic imaging agent in rats but no selective uptake in the pancreas was observed. DIFFERENTIATION OF SIDE CHAIN POSITIONAL ISOMERS OF AMPHETAMINE. W. H. Soine and M. N. Thomas*, Dept, of Pharm. Chem., MCV/VCU; R. E. Shark*, J. Scott and D. T. Agee*. Drug Examination Section, Bureau of Forensic Science, Consolidated Lab. of Virginia, Richmond, VA. The eleven side chain positional isomers of am¬ phetamine can be distinguished using .a combination of color tests, thin-layer chromatography and mass spectrometry. The primary amines, 1-phenylpropylamine and B-methylphenethylamine, exhibited chromatographic behavior similar to amphe¬ tamine but were readily differentiated using mass spectrometry. The mass spec¬ tra of N-methylphenethylamine was very similar to amphetamine, but using color tests and chromatography it was readily differentiated from amphetamine. THE VIRGINIA JOURNAL OF SCIENCE 155 RNA METHYLATION IN FREE AND BOUND POLYSOMES. Susan Spittle and Thomas 0. Sitz, Department of Biochemistry and Nutrition, Virginia Tech, Blacksburg, VA 24061. The late cytoplasmic 2 1 -0-methyl ati on of 5.8S rRNA is low in cancer cells and elevated in fully differentiated adult tissues. There is also a correlation of methylation with the amount of free and bound polysomes, i.e. the amount of bound polysomes is high in liver but low in hepatoma cells. It will be important to understand the functions of these late cytoplasmic modifications of ribosomal RNA and what impact they have on protein synthesis. We have developed a post-labeling procedure that will allow us to measure methylation levels in free and bound polysomes isolated from rat liver. This procedure and the results obtained will be presented. PHOTOLYSIS OF MONOSUBSTITUTED UNBRANCHED LONG CHAIN ALKYLCYCLOHEX- ANES. fofayne M. fit-al-ink. George W. Mushrush, and S. Patil, Dept, of Chemistry, George Mason University, Fairfax, VA 22030 Ultra-violet irradiation of long chain alkylcyclohexanes with a medium pressure quartz mercury vapor lamp for periods of 8 hrs to 6 days caused free radical decomposition. Analysis of the products was made by capillary gas chromatography. This showed the presence of most of the expected straight chain alkanes and 1-alkenes in ad¬ dition to their cyclohexyl counterparts that would normally be found from a random free radical decomposition mechanism. It is noted that higher molecular weight products are also produced in substantial amounts. These somewhat unexpected products are most likely produced by recombinations of the decomposition radicals present. (Supported by a George Mason University Research Grant) SOME PRESSURE-VOLUME-TEMPERATURE MEASUREMENTS ON LIQUID CRYSTALLINE ALKYL- AND ALKOXY-CYANOBIPHENYLS. Vincent J. Sullivan*, Glen C. Campbell* and Robert A. Orwoll, Dept, of Chem. , Col. of Wm. and Mary, Williamsburg, VA 23185. The melting of a thermotropic liquid-crystalline phase under the usual con¬ dition of constant pressure is examined in terms of a 2-step process: a phase transition at constant volume followed by an isothermal expansion. The total entropy change ASp is the sum of the entropy change ASy resulting from the disordering at the phase transition and the entropy change (DS/DV)-^ AV asso¬ ciated with the latent volume AV at the transition. Measurements of (9P/9T)y = (9S/3V)t and AV for alkyl and alkoxy derivatives of cyanobiphenyl lead to the conclusion that ASy is the smaller component of ASp at the nematic-isotropic transition of these compounds. (Supported by NASA Grant NAG-1-144 . ) ISOLATION AND RECONSTITUTION OF LOW DENSITY LIPOPROTEINS (LDL) : CONTINUING STU¬ DIES OF DELAYED-TYPE HYPERSENSITIVITY TO FOODS. Rosalie A. Tamburri* and Marga¬ ret F. Pinkston, Department of Biochemistry, Mary Baldwin College, Staunton, VA 24401. It is hypothesized that a defective plasma lipoprotein could serve as the sensitizing agent for delayed-type hypersensitivity (DH) to certain foods containing highly saturated triglycerides. In an attempt to identify the speci¬ fic causal agent, isolation of low density lipoproteins (LDL) from the blood serum of sensitive (test) and non-sensitive (control) persons was effected by precipitation procedures using heparin and Mn+2 and Mg*2, The LDL was then par¬ tially delipidated with heptane and reconstituted with known cholesteryl esters. Following reconstitution, the LDL of test and control were compared using iso- electricfocusing in foth agarose and polyacrylamide gels. The banding patterns of test and control showed distinct differences. It cannot yet be concluded that this difference is due to a defective apolipo protein, because further char¬ acterization of reconstituted LDL is necessary, but the evidence suggests that the research may be progressing in the right direction. Attempts will be made to incorporate saturated triglycerides as well as cholesteryl esters in the reconstituted lipoproteins. 156 THE VIRGINIA JOURNAL OF SCIENCE SUBSTITUTED CY CLOP ENTAD IENYL COMPOUNDS AS CARBOMETALLAT ION CATALYSTS. David W. Thompson and D. W. Moore*, Dept, of Chem. , Col. of William and Mary, Williams¬ burg, VA 23185. Bis(p^-cyclopentadienyl) titanium dichloride used with aluminum alkyls catalyzes efficiently the polymerization of alkenes and alkynes via a facile repetitive carbometallation reaction sequence. We have adapted this titanium-aluminum system to give a single, non-repetitive, carbometallation of the alkyne functionality in homopropargyl alcohols (eqn 1). However, the use of CH Cl A1 R Cl, + HOCH.CH.CECH — > Al.R ,C1, (OCH.CH.CECH) 2 x 6-x 2 2 0 2 x— 1 6-x 2 2 1. Cp2TiCl2 CH2C12 2. H HOCH CH 1 z\ C=CH / 2 R + HOCH_CH_ 2 2\ C / H H / C \ (1) the ring-unsub st ituted Cp2TiCl gives a mixture of reg ioisomers . We will pre¬ sent studies with the substituted cyclopentadienyl compounds, (CpMe) 2TiCl2> (Cp-t-Bu) 2TiCl2, and (CpMe5> 2TiCl2» and describe their effect on the distri¬ bution of reg io isomer s . APPLICATIONS OF MICROCALORIMETRY TO LIQUID /SOLID INTERACTIONS. J. P. Wightman, Chem. Dept., Va. Polytechnic Inst. & State Univ. , Blacksburg, Va. 24061. There are a number of experimental approaches to the study of interactions at the liquid/solid interface including thermodynamic, kinetic and spectroscopic which represent attempts to understand how molecules from the liquid state interact with solid surfaces. A powerful thermodynamic tool is calorimetry where heats of immersion (or wetting) are measured directly as the liquid wets the solid sur¬ face. In our laboratory, heats of immersion are measured in a Calvet micro¬ calorimeter. The operation of this calorimeter will be described briefly. Emphasis will be placed on the discussion of the heats of immersion determined in a number of systems including: (1) interaction of HCl(aq) and water with inorganic oxides; (2) interaction of organic liquids with coals; and, (3) inter¬ action of primer solutions and water with Ti and Ti alloy. (Research supported by MMRRI at Virginia Tech and the Commonwealth of Virginia.) PHENCYCLIDINE(PCP) ANALOGS : SYNTHESIS AND EVALUATION . R. L. Williams and James Hunt, Dept, of Chemistry, Univ. of Va., Charlottesville, VA 22901, and Dept, of Chem., Old Dominion Univ., Norfolk, VA 23508. As part of a continued search for a possible PCP antagonist, several novel phencyclidine analogs have been prepared and evaluated for possible agonist/antagonist activity in mice. This paper will discuss the synthesis and biological evaluation of these 4-phenyl -1 ,4di (1 -piperdinyl ) analogs. (1). R.RsCN R*CN, Rsi / (D THE VIRGINIA JOURNAL OF SCIENCE 157 D1 -2-PYRIDYLRETONE METHIODI DE : A NOVEL PYRIDYLATING AGENT. R. L. Williams and Sandra Neergaard, Dept, of Chem. Sci . , Old Dominion Univ., Norfolk, VA 23508. Di -2-pyridyl ketone methiodide (i) has been found to be an effective pyridylating agent with certain nucleophiles. The methiodide (1) can be selectively converted to the corresponding pyridyl esters, hydrazide and amide with the appropriate reagents. The reaction of (1) with other nucleo¬ philic species such as NaBH^, phenyl magnesi urn bromide and cyanide ion will be discussed. RsOR>NHNHJ,NHa NEW HETEROCYCLIC NEUROMUSCULAR BLOCKING AGENTS. R. L. Williams and Sandra Neergaard, Department of Chem. Sci., Old Dominion Univ., Norfolk, VA 23608. Recently several diaryl -4,5,6, 7-tetrahydro-imidazo-[4, 5, -C] pyridines (i) have been shown to exhibit neuromuscular blocking activity in mice. This work has now been extended to include the tryptamine ligand-and the corresponding iron complex (2) and (3). These new compounds appear to be depolarizing agents in contrast to the histamine analogs which have nondepolarizing neuromuscular blocking activity. This paper will discuss the synthesis and evaluation of these novel compounds. AN APPLE A DAY. John H. Wise, Dept, of Chemistry, Washington and Lee Univ. , Lexington, VA 24450. An APPLE microcomputer has been in use for nearly one year in the department. A variety of programs for demonstrations, calculations of laboratory reports, and tutorials have been developed and will be described. On-line data acquisition and treatment is planned for the future. SELECTIVE METHYLAT ION OF ENE-YNE-OLS VIA TITANIUM TETRACHLORIDE-TRIMERHYL- ALUMINUM. T. J. Zitzelberger* and D. W. Thompson, Dept, of Chem., Col. of William and Mary, Williamsburg, VA 23185. We have shown that y-alkynols are selectively carbometallated with a TiCl^-AlMe3 reagent system accoding to the following reaction sequence (eqn 1). CH Ci Al.Me, + HOCH-CH.CHCH — „. > Zb Z Z 0 1. TiC14 -> HOCH CH s Jl /C - \ ■78 H CH„ ch2ci2 2. H"1 A1 Me5(OCH CH CECH) (1) The methylation occurs with a wide variety of alkynols at the carbon furtherest from the hydroxyl group; the carbometallation occurs in a syn fashion. This reaction sequence has now been extended to ene-yne-ols of the type HOCHCH CEC-CCH ^R 2 R' 2 R - H, C H ; R' = H, CH 158 Education Section THE VIRGINIA JOURNAL OF SCIENCE INDIVIDUALIZED CONTRACTUAL EVALUATION FOR BIOLOGY STUDENTS. H. _S. Adams, D. S. Lancaster Cmnty. Col., Clifton Forge, VA 24422. Grade distribution of 349 gen¬ eral biology students (during a six-year period) using traditional evaluation methods was compared with that of 281 general biology students (during a sepa¬ rate six-year period) using a student-instructor contractual agreement. For the contract agreement, each student was allowed to select both the activities and percent weight of each activity used in deciding their grade. Success rate of students (i.e., those attaining at least "C") using the contract approach increased by nine percent (from 70% to 79%) over those using traditional evalua¬ tion for the six-year period. During the final three years of using the con¬ tracts, rate of success in biology averaged 85%. Among those surveyed, students liked the opportunity to (1) de-emphasize exams as sole factor in determining their grade and (2) selecting those activities on which their grade is based. CONTRASTING AB AND DF BIOLOGY STUDENTS. H.S. Adams, A.M. Dutrow*. and K.S. Bowers*, D.S. Lancaster Cmnty. Col., Clifton Forge, VA 24422. Twenty-one variables were compared for AB and DF general biology students. The only vari¬ ables for which the groups did not differ significantly (alpha = .01) were age and length of time elapsed from high school graduation before taking the course. For all other variables, including high school grades, rank in graduating high school class, and CGP (Comparative Guidance and Placement) scores, the AB group was significantly higher. Discriminant analysis revealed misclassif ication of five DF and eight AB students. Means for misclassif ied DF students were consid¬ erably lower in high school rank and CGP motivation, math interest, and biology interest scores than those for the AB group with which they should have been classified. The AB students who were predicted to have received D or F differed from the DF group by averaging higher on CGP reading, sentence, and math scores. NATIONALLY DEVELOPED ENVIRONMENTAL EDUCATION PROGRAMS RECENTLY AVAILABLE IN VIR¬ GINIA. TERESA M. AULDRIDGE*, VA DEPT. OF EDUCATION, BOX 6Q, RICHMOND, VA 23216 AND BRENDA A. BAKER*, HENRICO CO. SCHOOLS, P. 0. BOX 40, HIGHLAND SPRINGS, VA 23075. Project Learning Tree and Project WILD were both initiated by the West¬ ern Regional Environmental Education Council. They are interdisciplinary en¬ vironmental education programs intended to supplement the regular curriculum grades K-12. The seven major principles of PLT focus on the forest environment and our use of its resources, while WILD emphasizes the ecology and management of wildlife. A 1981 grant from the American Forest Institute made PLT available free to any educator in Virginia who attends a workshop. To date, over 1,700 teachers have received the materials. Project WILD is being funded jointly by the VA Commission of Game & Inland Fisheries and the Izaak Walton League. Vir¬ ginia was one of three states in the nation to participate in field testing the rough draft of WILD during the 1982-83 school year. Final versions will be available for educators who attend workshops beginning in October 1983. CRYSTAL WORLD - ITS USE AS A CLASSROOM. R. Wesley Batten, Director, Science Presentations Division, Science Museum of Virginia, Richmond, Virginia (23220). The world class exhibition at the Science Museum of Virginia on the science of crystallography is a rich instructional resource for illustrating difficult concepts in science. Crystal World may be used as an activity base and as a project model for elementary and secondary science. Crystal chemistry, states of matter, properties of metals, atomic structure, symmetry, physics of light, refraction, and mineral identification are but a few of the topics that stimulate students' understanding of basic scientific truths. THE VIRGINIA JOURNAL OF SCIENCE 159 CREATIONISM THROUGH THE BACKDOOR: THE CASE OF LIBERTY BAPTIST COL. Michael L. Bentley, Dept, of C.& I., Sch. of Ed., Univ. of Va., Charlottesville, VA 22903. On Dec. 10, 1982 the Va. Board of Ed. provisionally approved the biology program for the preparation of ; public school teachers at Liberty Baptist College, Lvnchburg, Va., despite the protest of the ACLU, Va. Academy of Science and others I that the college's program taught creationism as science. Va. certification also extends automatically to 34 other states. The LBC program was subjected to the standard review procedure for programs seeking approval for teacher education. The criteria for approval were formulated before creationism became a signifi¬ cant issue in science education. Only one scientist was involved in the procedufe. The criteria for approval and the incut of the scientific community in such cases need a second look. Even educa- ; ted laymen do not apparently appreciate the scientific issue involved in teaching creationism as science. The situation is evidence of the need for more emphasis in education on the philos¬ ophy of science. : THE ROLE OF THE FAMILY IN THE PROMOTION OF SCIENCE LITERACY. Elizabeth A. | Cornell*, VA Inst, of Marine Science, Sch. of Marine Science, Col. of William | and Mary, Gloucester Point, VA 23062. Final results from a NSF-funded series I of studies being conducted by the Smithsonian's Chesapeake Bay Center for I Environmental Studies and the VA Inst, of Marine Science will be presented. I The series includes characterizing the families visiting science centers, measuring participation rates and persistence of family members whose young¬ sters bring home "take-home, family activities" from school, characterizing what makes a good family-learning situation, and how parents and children teach j and learn from each other. Theories and models of adult education and the role of the family in informal learning are discussed. Concepts are drawn together from a wide variety of disciplines (e.g., psychology, sociology, museum educa¬ tion, higher education) . FORMATION OF ECOLOGICAL CONCEPTS BY UPPER ELEMENTARY STUDENTS. Julia H. Cothron, Hanover County Sch., Ashland, Va. 23005, & Ertle Thompson, Sch. of Ed., Univ. of Va., Charlottesville, Va. 22903. SCIS students, grades 4-6, completed a multiple-choice and free-sort task. Grade and ability, not sex, influenced concept attainment and similarity of conceptual system to discipline structure. Sequence was not influenced by grade, ability, sex. Concept attainment was promoted by conceptual system similar to discipline structure. Sixth graders recognized life requirements, interactions via food, mineral, gas exchange, and community groups, but not attributes of higher-order concepts: producer, consumer, decomposer, environmental factors. Discipline experts, not students, related examples to higher-order concepts. Recommendations for K-6 curriculum were: (1) promote conceptual system similar to discipline structure, (2) base concept maps upon lower-order concepts or attributes, (3) eliminate higher- order concepts, (4) emphasize interactions of plants, animals, decayers, and materials j feeding relationships » and life requirements. CHEMICAL WASTE IN SCHOOLS. Timothy W. Cotman*, Va. Dept, of Education, Box 6Q, Richmond, VA 23216. This seminar will document the efforts of the Science Ser- I vice of the Virginia Department of Education, in conjunction with several other agencies, to purge unwanted and excessively dangerous chemicals from public j schools in Virginia. The need for this action will be examined in view of the | importance of the laboratory aspect of secondary chemistry courses, regulatory considerations, practical implementation and the actual mechanics of the present project. 160 THE VIRGINIA JOURNAL OF SCIENCE POTENTIAL OF THE VIDEODISC IN INTERACTIVE INSTRUCTION. Mark Delp,* Division of Instructional Media and Technology, Virginia Department of Education, Richmond, Virginia 23216. The impact of employing interactive high technolo¬ gies to truly individualize instruction is just beginning to be acknowledged. Up to this time, the interactive technologies most employed in instruction (computers and most currently microcomputers) have been limited to the presentation of alphanumerics , graphics and sound, primarily because of mass storage limitations. The optical laser read videodisc makes the integration of full color motion and stereo sound with microprocessor display generation and control a reality. Three levels of interactivity are possible and will be demonstrated in this session using a laser reflective optical videodisc player with built-in microprocessor. A SYSTEMS APPROACH TO DEVELOPING EFFECTIVE PROGRAMS OF SCIENCE. Joseph Exline, Associate Director of Science, VA Department of Education, P. 0. Box 6Q, Rich- mond, VA 23216. Accepting the fact that criteria have long been available for identifying exemplary programs of science and that valid research has long been available for pinpointing the shortcomings in science programs, it is contended that the major problem in providing a quality science program has to do with the lack of effective program implementation on a large scale and with a long range commitment. The crucial first step to improving efforts in science education is the development of a sound long range plan. Developing a systems approach im¬ plementation plan is the crucial second step to improving effectiveness in sci¬ ence education and is the step which has been most neglected. The systems approach plan must consider at least the following: teacher background, admini¬ strative support, learning environment, instructional materials, facilities and equipment, and community support. AGRICULTURE IN THE CLASSROOM. Joyce H, Jones, Dept, of Poul . Sci., Va. Poly¬ technic Inst, and State Univ., Blacksburg, VA 24061. Recent studies by the American Farm Bureau have determined that public school teachers in both urban and rural settings rely on classroom textbooks for teaching information relating to agriculture. Frequently, classroom textbooks and library resource materials present agricultural information that is outdated and/or inaccurate. During the last twenty years, Agricultural research has produced numerous improvements in plant and animal agriculture. These include improvements through genetics, physiology, nutrition, management, and/or animal behavior research. Agricultural research and resulting improvements in production, quality and cost factors of agricultural commodities can become educational classroom resource tools to introduce students to a) the scientific method and b) how research results can affect daily lives. Science and Technology in the Poultry Industry has been presented in teacher workshops by Cooperative Exten¬ sion Service Specialists and Agents. Follow-up teacher evaluations cite the availability of accurate agricultural information and resulting classroom dis¬ cussions and activities (i.e., simple experiments) as generating positive student understanding of the necessity for research and an appreciation of the problems faced by agricultural commodity farmers. THE VIRGINIA JOURNAL OF SCIENCE 161 THE CHANGING NATURE OF MARINE SCIENCE EDUCATION. Harris B. Stewart, Jr. , Ctr.for Marine Studies, Old Dominion Univ. , Norfolk, V a. 23508 Susanne S. Jackman, Applied Marine Res. Lab., Old Dominion Univ., Norfolk, Va. 23508. The teaching of marine science at our schools and colleges is changing as man's perception of his relationship to the marine environment changes. The eco-hysteria of the 1960's and 1970 ' s resulted in environmental approaches and policies that are now being reevaluated in the light of new knowledge and of further consideration of their socio-economic implications. Educators at all levels must develop their own means of keeping abreast of the advances in our scientific knowledge of the sea and more particularly in the changing public attitudes towards the sea, its resources, its legal regime, and its interactions with the land. Courses called "marine science" or "oceanography" will need to be expanded to include such marine aspects as the legal and the economic ones. Coastal zone management, resource recovery, marine technology, and international aspects might well be covered in addition to the traditional fields of marine biology, chemical, geological and physical oceanography. WHO OWNS THE BIOLOGY CURRICULUM? Wayne Moyer, National Association of Biology Teachers, 11250 Roger Bacon Drive, Res ton, VA 22090. The textbook is the curriculum for most teachers; but who owns the textbooks? The Federal Government is out of the curriculum writing business, perhaps for good. Com¬ mercial publishers are under pressure to modify their texts to meet various demands. Local school authorities tend to concentrate curriculum responsi¬ bility in the hands of administrators, treating teachers like hired hands. Who then is responsible for what is taught in biology classes? This presenta¬ tion will review trends in biology texts as seen in examples printed during the last 60 years, while examining the question, "What is basic in biology?" Human ecology seems to be emerging as the answer for the 80s. The role of classroom teachers in curriculum development will be considered, and includes: (1) cultivating curiosity about the natural world, (2) accurately transmit¬ ting our scientific heritage, (3) departing from the textbook to encompass other authorities, and (4) reclaiming the teacher’s academic birthright to write curricula. SEA GRANT IN VIRGINIA: WHAT IT’S ALL ABOUT. William L, Rickards, Virginia Graduate Marine Science Consortium, University of Virginia, Charlottesville, VA 22903 The Virginia Sea Grant Program is an inter-institution, inter-disciplinary program of applied research, education and advisory activity designed to foster the wise development and management of marine and coastal resources. Research projects cover such diverse subjects as improved techniques for farming oysters and clams, the biology of commercially exploited organisms, improved weather forecasting for Chesapeake Bay, uses for seafood processing wastes, and anti¬ cancer substances from marine animals. The Education portion of the Program emphasizes educating teachers through workshops, information dissemination systems, and field activities as well as the development of marine 4-H educational materials. Through the Marine Advisory Services (MAS) , technical information is transferred to the private sector and state agencies on subjects including commercial and recreational fishing, marine trades and businesses, seafood processing and marketing, and aquaculture. 162 THE VIRGINIA JOURNAL OF SCIENCE THE GOVERNOR'S SCHOOL INTEREST CENTER Betsy M. Waring, Mathematics and Science Center, 2401 Hartman Street, Richmond, VA 23223 The Governor's School Interest Center is a non-residential nrogram, funded by the Department of Education, for identified gifted students in 10 counties, four cities and accredited private schools in the Richmond area. The Mathe¬ matics and Science Center developed the pilot program for the State in summer 1982. In summer 1983, the Mathematics and Science Center will be one of three centers, and it will offer the only Interest Center program in science and mathematics for gifted students in grades 8-12. In a Monday through Friday format, students explore a topic through lec¬ tures, discussions, laboratory investigations, field work, and with visiting consultants . In summer 1983, interdisciplinary programs in fresh water environments (grades 8-9) and marine environments with emphasis on Chesapeake Bay (grades 10- 11) will include camping and on-water experiences. An interdisciplinary chem¬ istry of living systems course will include visits to medical and industrial re¬ search centers. All courses will require project work. Engineering Section TECHNICAL SYMPOSIUM. R.D. Kirchner, Presiding. The Engineering Section of the Virginia Academy of Science devoted this year's meeting to discussions of the current crisis in America's science education programs, its effect on the national defense and economic prosperity, and what can and must be done to remedy the situation. Prominent speakers from government, industry, and educ¬ ational institutions presented their views on various aspects of this vital issue and outlined actions that must be taken to insure the numbers of gualified engineers, scientists, and technical workers needed to compete in the growing worldwide technical revolution. Speakers included Dr. Bernard Kulp, Air Force Systems Command; Bryce Jewett, Jewett Machine Manufacturing Co.; Margaret Marston National Commission on Excellence in Education; Dr. Marvin Cetron, Forecasting International Ltd.; Dr. John Casteen, State Secretary of Education; Bill Aldridge National Science Teachers Association; Dr. J. Wade Gilley, The Institute for Science and Technology; Dr. Rodney Hannaman, Reynolds Metal Co.; Nancy Flinn, American Chemical Society; and Pamela Kurstedt, Virginia Polytechnic Institute and State University. Environmental Science Section ZOOPLANKTON POPULATION DYNAMICS OF LAKE ANNA, VIRGINIA, 1973-1982. John B_. Bailey, Environmental Services Department, Virginia Electric and Power Company, Richmond, Virginia 23261. In 1972, Virginia Electric and Power Company impounded the North Anna River to create a 3887 hectare reservoir to provide condensor cooling water for its North Anna Nuclear Power Station. Zooplankton sampling occurred both before (1973-1975) and after (1978-1982) power generation was initiated in 1978. Comparisons of zooplankton density and diversity showed no significant differences between pre and post-operational populations in the reservoir. THE VIRGINIA JOURNAL OF SCIENCE 163 TRANSPLANT EXPERIMENTS WITH SUBMERSED AQUATIC VEGETATION IN THE POTOMAC RIVER. Nancy Bartow* and Virginia Carter*, U.S. Geological Survey, Reston, Va. , 22092. The tidal Potomac River has been nearly devoid of submersed aquatic vegetation since the 1930's. Transplant experiments were conducted at four sites during 1980-1982 to determine whether Vallisneria americana will survive in the tidal Potomac River under present conditions. Exclosures were placed around several transplant beds to test the effect of grazing. At three out of four sites, transplant beds protected by exclosures during the first growing season were successful and regrew the following year. Unprotected plants disappeared or were clipped off by unidentified browsers. Site characteristics play a role in transplant success. Of the characteristics measured (fetch, exposure, sub¬ strate, light penetration, sediment nutrients, and heavy metals), substrate and light penetration seem to be most important. Sedimentation from storm events may have been a factor in the elimination of submersed aquatic vegetation. A laboratory study was conducted to investigate the effects of sediment deposition on the emergence of Vallisneria from tubers. About 65 percent of the Vallisneria emerged and grew green leaves when buried in 15 cm of sediment, about 25 percent emerged and grew green leaves when buried in 20 cm, and none emerged when buried in 25 to 55 cm of sediment. WATER ELEVATION AND SALINITY SURVEY OF LONDON BRIDGE AND WEST NECK CREEKS. Carvel Blair, Stephen Danna*, and Joung Kim? Department of Oceanography, Old Dominion University, Norfolk, VA 23508. Over a one-year interval (February 1982- January 1983) we surveyed water elevation and salinity in the London Bridge-West Neck Creek system. Stations were spaced along the 20-mile stretch from Lynnhaven Inlet (on Chesapeake Bay) to the North Landing River (on the Intra-Coastal Waterway). The upper limit of salinity intrusion varied between 6 and 13 miles upstream from the inlet. Previous studies had found an inverse correlation between salinity and 30-day rainfall. We found the correlation to be much stronger with rainfall during the previous 2 or 3 days than with the previous month. We also found a tidal change in water elevation extending an average of about 10 miles upstream, somewhat farther than the mean salinity intrusion. Celerity of the tidal wave was about 5 cm/sec. A SURVEY OF ACTION AND ATTITUDES TOWARD URANIUM MINING AND MILLING IN VIRGINIA. Margaret A. Brenner & Joseph Norris, Dept, of Biol., Lynchburg Col., Lynchburg, Va. 24501. The Marline Uranium Corp. announced on July 21, 1982 that a large deposit of uranium oxide (U3OQ) had been discovered in the Triassic Basin region of south central Virginia. Marline's work in the area of Pittsylvania County has included surface studies and exploratory drilling procedures. Detailed obser¬ vations of the geology of the area and a proposed plan for the mine/mill complex have also been compiled. This presentation will explore both the optimistic and pessimistic views of uranium mining and milling by exploring governmental and public response to Marline's plan, and also parovide a detailed description of Marline's work thus far and their plans for the future. 164 THE VIRGINIA JOURNAL OF SCIENCE ACID RAIN MONITORING IN VIRGINIA. Arthur L. Buikema, Jr_. and Boris I. Chevone, University Center for Environmental Studies and Department of Biology and Air Pollution Laboratory and Department of Plant Pathology and Physiology, Va. Polytechnic Institute and State University, Blacksburg, Va. 24061. The Virginia Acid Precipitation Network (VAPN) was established in late 1980 as part of the research activities carried out under the direction of the Technical Advisory Committee to the State Air Pollution Control Board. The purpose of this research is to provide a data base for determining the extent, significance and possible source contributions of acid deposition in Virginia. Since 1982 there are 8 stations in the network. Rain samples are monitored weekly for pH and conductivity. If sufficient sample exists, the concentrations of sulfate, nitrate, chloride, ammonium, calcium, magnesium, sodium and potas-sium ions are also measured weekly. Two sites are monitored weekly for arsenic, selenium, manganese, vanadium and aluminium. Quarterly all sites are monitored for 19 trace elements. THE EFFECTS. OP DREDGE SPOILS FROM THE' ELIZABETH RIVER OK THE CA LA NO ID C0PEF0D ACARTIA CLAUSI Renee S. Crpuch Dept, of Biol. Sci. Old Dominion Univ. Norfolk VA 23508. A 96-hDur, static, suspended particulate bioassay was conducted to assess the effects of dredge spoils from a highly industrialized region of the Southern Branch of the Elizabeth River, VA, on the copepod Acartia clausi . The mortality in all test containers was highly significantly different from contrdLs (p<.0l) beginning at the L5-hr time interval. The 48-hr and yC-hr LC50’s were 13. 18/s and 12.59> suspended solid elutriate, respectively. The incipient LC50, that which produces $0% nortality over an infinite time, was elutriate. The suspended solid elutriate from this region contained high levels of ammonia, high concentrations of heavy metals, and a high silt-clay content. The observed lethal effects may be due to some toxin or combination of toxins, whether am¬ monia, heavy metals or unmeasured contaminant, as well as physical stress from suspended particulates, found in this heavily industrialized region of the Eliz¬ abeth River. Additional research is4 needed to thoroughly assess the impact of dredge spoils on the zooplankton community. THE EFFECT OF SEA NETTLE ABUNDANCE ON OTHER TROPHIC LEVELS IN THE LOWER CHESA¬ PEAKE BAY REGION. Michael G. Kelly* & Dr. David L. Feigenbaum, Dept, of Oceanography, Old Dominion Univ., Norfolk, VA 23508. The abundance of the medusoid stage of the sea nettle, Chrysaora quinquecirrha and other gelatinous and crustacean zooplankton and phytoplankton were investigated at four stations in the Lafayette and Elizabeth Rivers, Virginia, from May to October 1982. The sea nettle, when abundant, directly or indirectly controls much of the life in the water column through its feeding activities. Medusae appeared in early June and the ctenophore, Mnemiopsis leidyi, sharply declined. The crustacean population significantly decreased in the presence of M. 1 ei dyi but subsequent¬ ly increased when the sea nettle reduced the ctenophore population. The crus¬ tacean abundance was variable through the second half of the study. Food was not a limiting factor for the crustaceans. A MATHEMATICAL MODEL STUDY OF WATER QUALITY OF A SMALL TIDAL CREEK. Albert Y. Kuo and Stephen Williams*, Va. Inst, of Marine Sci., Col. of William and Mary, Gloucester Point, Va. 23062. A one-dimensional mathematical model has been developed for use in small tidal streams to investigate the short-term intra- tidal and diurnal fluctuations as well as the long-term seasonal variations of water quality. The model solves the continuity and momentum equations simul¬ taneously through the use of a semi-implicit finite difference scheme. The solutions of these equations provide water velocity and surface elevation, which are then used to solve the mass-balance equation. The solution of the mass-balance equation describes the longitudinal and temporal distributions of eight dissolved or suspended substances comprising the ’ecosystem’ . The model has been calibrated and validated using data from the Little Hunting Creek, a small tidal stream joining the Potomac River. The model has also been used to investigate the cause-effect relationship controlling water quality conditions in the Creek. THE VIRGINIA JOURNAL OF SCIENCE 165 SEASONAL COMPOSITION OF PHYTOPLANKTON IN THE ELIZABETH RIVER AND HAMPTON ROADS AREA OF VIRGINIA. Richard Lacouture* , and H. G. Marshall. Depts. of Biological Sciences and Oceanography, Old Dominion University, Norfolk, Va. , 23508. Monthly collections in Hampton Roads and the Elizabeth River were collected for phytoplankton analysis over a 13-month period. Maximum concentrations occurred in spring and summer, decreasing into fall with lowest numbers in winter. Phytoplankton biomass levels were greatest in early spring and early summer, with fall having the lowest amount. Concentrations were generally greater or comparable in. the Elizabeth River when compared to other stations sampled in the lower Chesapeake Bay. With the exception of early spring, the phytoplankton biomass was generally less than what was found in the lower Bay. The phytoplankton also contained an abundant ultraplankton component composed mainly of cyanophycean and chlorophycean species. The dominant species through¬ out the year included Skeletonema costatum, Leptocylindrus minimus , L_. danicus , Asterionella glacialis , Prorocentrum minimum, Cryptomonas spp . , and Emiliania huxleyi . The spring populations were dominated by S_. costatum with a shift to a combination of ultraplankton and phytof lagellates more prominent in the summer maximum. SEASONAL COMPOSITION OF PHYTOPLANKTON IN THE LOWER CHESAPEAKE BAY REGION. Richard Lacouture*, and H. G. Marshall. Depts. of Biological Sciences and Oceanography, Old Dominion University, Norfolk, Va., 23508. Monthly observations were made at seven stations in the lower Chesapeake Bay for a 13-month period. One hundred ninety-six species were identified with diatoms, dinof lagellates , and an unidentified ultraplankton component the most prominent species. Spring was dominated by small, chain forming diatoms (Skeletonema costatum, Rhizosolenia f ragilissima, Thalassiosira nordenskioeldii) and an ultraplankton assemblage <3 ym. Prorocentrum minimum and other dino- f lagellates became more abundant in summer along with several Chaetoceros spp. During fall, there was a general increase of the smaller diatoms (S_. costatum, Leptocylindrus minimus, Thalassionema nitzschioides) and cryptomonads. A flora similar to the fall assemblage continued into winter, but in lower concentra¬ tions. Ubiquitous throughout the year were ultraplankton cells, <3 ym in size, that were mainly composed of cyanophycean and chlorophycean species. Maximum cell concentrations occurred during early spring and mid-summer. Highest phytoplankton biomass levels were in spring and early fall, with the lowest biomass recorded in mid-summer and during winter. SEASONAL CELL VOLUME AND CONCENTRATION PATTERNS FOR MAJOR PHYTOPLANKTON GROUPS OVER THE NE CONTINENTAL SHELF. H. G. Marshall, L. Jugan* , C. K. Rutledge*, P. Zimba* , and D . A . Randolph* . Department of Biological Sciences , Old Dominion University , Norfolk, Va. 23508 . Analysis of a data set consisting of 714 station samples from 26 cruises between 1978 and 1982 indicated large scale distribution patterns for the phytoplankton . Areas of high biomass concentrations had closer direct relation¬ ships to cell concentrations near the major bay systems and Georges Bank, and over the shelf during seasonal periods of peak development . The seasonal appearance of large sized diatoms and dinof lagellates enhanced the biomass levels during winter and summer. A seasonal progression of increased develop¬ ment from early spring through summer was indicated from the Gulf of Maine southwest coast seaward to Georges Bank. Ubiquitous species over the shelf included Asterionella glacialis, Guinardia f laccida, Leptocylindrus danicus , Rhizosolenia alata, Skeletonema costatum, Thalassionema nitzschioides, Cerat ium fusus , Ceratium lineatum, and Ceratium tripos . Supported in part by the NOAA/NEMP Ocean Pulse program. 166 THE VIRGINIA JOURNAL OF SCIENCE EFFECT OF SHELF ENVIRONMENT ON YEAR CLASS STRENGTH OF ATLANTIC CROAKER (Micropo- gonlas undulatus) . Brenda L. Norcross, Herbert M. Austin, VA Inst. Mar. Sci. , Sch. Mar. Sci., Col. Wm. & Mary, Gloucester Pt., VA 23062, and Sharon K. LeDuc*, NOAA/NESDIS/ASISC , Columbia, MO 65201. The geographic center of croaker spawning varies in relation to shelf bottom temperatures. These interannual fluctuations in bottom temperatures appear to be linked to timing of seasonal cessation of the southerly wind component resulting in variation in onset and northward placement of croaker spawning. Normal and anomalous wind regimes drive transport mechanisms which affect larval croaker, and therefore signifi¬ cantly impact recruitment of juveniles to the Chesapeake Bay. The effect of wind- driven transport is positive or negative depending on larval position in the water column. Year class strength of Atlantic croaker is strongly dependent upon the coherence between the seasonal wind shift, timing and area of spawn, wind driven transport, and position of larvae in the water column. USE OF NET BARRIERS TO EXCLUDE STINGING JELLYFISH FROM SWIMMING BEACHES. A. J. Provenzano, D. L. Feigenbaum, C. Blair, D. F. Johnson*, and M. Kelly*, Dept, of Oceanography, Old Dominion University, Norfolk, VA 23508. Jellyfish, including the stinging nettle, Chrysaora quinquecirrha, are abundant in summer in Chesa¬ peake Bay. Their presence greatly curtails recreational use of the beaches. Tourism, local business and real estate values are all adversely affected. Ex¬ perimental jellyfish exclusion zones were established at two public beaches in Norfolk in mid-summer, 1982. An attempt was made to determine technical feas¬ ibility of such barriers, the advantages of various netting materials, public acceptance of protected areas and the projected costs of larger scale enclos¬ ures. At one site, structural damage to rigid net frames prevented gathering of adequate data on design. At the second site, flexible netting hung from temporary pilings reduced jellyfish abundance within the enclosed area. Public utilization of protected beach was many times higher than adjacent unprotected beach. Seining and flushing of jellyfish out of the enclosure on tidal cur¬ rents reduced impact of jellyfish which penetrated the barrier. Design modifi¬ cations, economic studies and season-long feasibility testing of a larger unit are planned for 1 983. ALTERATION OF THE TREE GROWTH TO CLIMATE RELATIONSHIP IN RED SPRUCE (PTCEA. RUBENS SARG.). L. J. Puckett, U.S. Geological Survey, Reston, Va. 22092. Incremental growth cores were collected from red spruce (Picea rubens Sarg.) trees growing on bedrock and on glacial till at the Hubbard Brook Experimental Forest. Results of regression analyses of tree-growth indices against orthogonally transformed tem¬ perature and precipitation data for two periods, 1932-56 and 1956-80, indicate that the relationship of tree growth to climate has been altered. For tree grow¬ ing over bedrock the responses to climate and to prior growth have decreased. For trees growing over till the response to climate decreased and the response to prior growth increased. Statistical t-tests and F-tests of the climate data of the two periods indicate that climate has been essentially the same. Droughts have occurred in 60 percent of the last 50 years with a peak magnitude in the mid-1960's. It is suggested that toxic Al^+ generated through dissolution of reactive alumina via hydrogen ion neutralization reactions in glacial till soils may be damaging the root systems of red spruce. However as a result of organo- metallic complexes formed in the humus, roots there may not be affected, effect¬ ively restricting root systems of trees growing on till to the humus layer. The root systems of trees growing in humus over bedrock would not be so negatively affected . THE VIRGINIA JOURNAL OF SCIENCE 167 THE INFLUENCE OF FOREST CANOPIES ON PRECIPITATION CHEMISTRY IN NORTHERN VIRGINIA L. J. Puckett, U.S. Geological Survey, Reston, Va. 22092. Throughfall was colle¬ cted in adjacent coniferous and deciduous stands at the Mill Run Research Water¬ shed in northwestern Virginia. Nine sample sets covering a period of 13 weeks between July and November had mean pH values of 4.13, 4.23, and 3.92 for precip¬ itation, deciduous throughfall, and coniferous throughfall, respectively. Beneath both canopy types there was a net gain of H+, Ca^+, Mg^^NO^-, and SO^- ions and a net loss of NH^+ ions. In precipitation H 2^^ NH^ provided 71 per ■. cent of the cation charge. ^neath conifers H and Ca provided 68 per cent and beneath hardwood H ^ Ca , and K provided 83 per cent of the cation charge. SO^ , N0^ , and Cl provided 53, 21, and 25 per cent, respectively, of the an¬ ion charge in precipitation. Of the anions in throughfall SO^ , NO^ , and Cl provided 57, 30, and 12 per cent, respectively, beneath conifers+and 64, 21, and 14 per cent, respectively, beneath hardwoods. The net gain of H was 95 per cent beneath conifers and 8 per cent beneath hardwoods. SO^ and NO3 ions increased 136 and 209 per cent, respectively, in conifer throughfall and 86 and 51 per cent, respectively, in deciduous throughfall. These results indicate the importance of dry deposition to tree canopies in evaluating potential impacts to forest ecosystems. OCEAN TIDE RESEARCH AND THE BRAKING OF THE EARTH ROTATION. Ernst W. Schwiderski, Naval Surface Weapons Ctr., Dahlgren, Va. 22448. The paper highlights the centuries' old and colorful history of ocean tide research, which culminated in an ocean tide model that allows a prediction of the water height with an accuracy of better than 10 cm anytime and anywhere in the real world oceans. The constructed tidal waves rotate around fixed points over huge areas in about 12 hours. Among the broad range of vital applications, the mechanism of tidal friction and braking of the earth rotation is explained in some detail. It is shown that while the moon pulls the earth and ocean tides around, the oceans gear into the moving ocean and brake the rotation of the earth, burning about 53% of the tidal energy (ca. 3.56TW for M„) into wasted heat. A more specific paper on "Hydrodynamical Mapping of Global Ocean Tides" will be presented in the session of Astr. , Math., and Physics. AMOUNTS OF PARTICULATES PRESENT IN ^HE ATMOSPHERE OF LVNCHBURG, VA. Randee Stenroos^f-, Lynchburg, Col., Lynchburg, Va. 24503. To find the level of particles present in the atmosphere of Lynchburg Va. pieces of tape were placed in various areas around the city. The tape was left ouffor 10 hours. It was then collected, placed on microscope slides, and the particles present were counted. Pictures of the particles were made with a Lutz- Ortholux microscope which enabled the author to see the variety of particles present* These results showed: the heaviest concentration in the backyard of a house inside the city. There were roughly 500 particles on the pieces of tape 2 centimeters by 2-g- centimeters. The next area of greatest concentration was the woods inside the city limits. There were roughly 400 particles present on pieces of tape collect? ed. The area of lowest concentation was right inside the city limits. The number of particles present were roughly 300 particle PROPOSED ALTERNATIVES FOR LIMITED ACCESS TO FALSE CAPE STATE PARK THROUGH BACK BAY NATIONAL WILDLIFE REFUGE. Mary Ann Sykes. i k Lynchburg Col., Lynchburg, Va. 24501. In order to meet recreational needs identified in the Va. Outdoors Plan 1979, the Commonwealth of Va. is seeking limited access through Back Bay National Wildlife Refuge. At present, the park is accessible only by boating, biking, or hiking. Various proposals have been developed to obtain this limited access through the refuge. It is thought that the present policies and object¬ ives of both the refuge and the park can be maintained by the exchange of land or right-of-way grants. These proposals are reviewed. 168 THE VIRGINIA JOURNAL OF SCIENCE Preliminary Studies of Selected Heavy Metals in the Waste Waters of the Lynch¬ burg Area in Central Virginia. Walter Younger,* Regional Waste Water Treatment Facility, Lunchburg, Va. 24505. Analysis of 9 metals (Pb, Cd, Zn, Cr, Fe, Ni, Mn, Cu, & Ag) were performed on the Waste Treatment Facility’s influents, settling tank overflows, efflent, and waste sludges and also on 6 local streams and the James R. to determine typical, metal levels. The Facility influents are heavily contaminated in comparison to the Lynchburg streams and the river. Metal removals from liquid portions by quiescent settling were poor except for Pb. When properly operating relatively large amounts of metal are removed by accumulation in the biomass except in the case of Mn which showed 0% removal. Concentration of toxic metals in sludges is believed to be a cause of concern. To illustrate the potential danger, grass and cherry tomatoes grown- in some aged sludge (excess 10 yrs) were compared with grass grown in uncontaminated soil. High metal levels were observed in sludge grown grass but levels in tomatoes were comparable to the control grass. Implications are that fruit producing ^plants may be safe to grow in sludge soil but caution is recommended in utilization of leafy type plants. Geology Section RE CRYSTALLIZATION FEATURES IN DEFORMED ANTIPERTHITIC PLAGIOCLASE , MONTPELIER ANORTHOSITE, HANOVER CO., VA. , Elizabeth L. Beasley*, Col. of William and Mary, Williamsburg, Va. 23185. Recrystallization features in deformed antiperthitic plagioclase of the Montpelier Anorthosite are related to deformation and hydr¬ ation. Antiperthitic plagioclase, the dominant mineral and textural feature in the anorthosite is modified by recrystallization. As recrystallization occurs, irregular strips and patches of epitaxial remobilized plagioclase are formed. Albite and pericline deformation twins suggest that strain enhanced diffusion was responsible for the remobilization of the plagioclase. Myrmekite in the Montpelier anorthosite is unique for it occurs between non-perthitic plagioclase and antiperthite. In accord with the ideas of Becke, it is believed that the replacement of the potash feldspar component of antiperthite by non-perthitic plagioclase causes the generation of silica. The dissolution of potash feldspar lamellae near non-perthitic plagioclase grain boundaries and their association with quartz droplets is support for this interpretation. The generated silica accounts in part for the quartz content of the myrmekite and poikilitic plagioclase. THE MID-PROVINCE STRUCTURAL FRONT BETWEEN FREDERICK CO. AND BUCHANAN, VA. : AN INTERPRETATION. Kenneth F. Bick, Dept, of Geol. , College of William and Mary, Williamsburg, Va. , 23185. The leading edge of the North Mountain-Staunton- Pulaski fault complex and discontinuous fold systems immediately west mark the Mid-Province Structural Front. Whereas the Allegheny Front essentiallly reflects faults ramping from Rome Fm. to Martinsburg Fm. , the Mid-Province Front is thought to be the step-up of a sole fault in the Martinsburg, associated with less significant Rome to Martinsburg ramping expressed as the fold system; Front thrusts branch from the Martinsburg sole fault. Assumptions involved in serial cross sections constructed to test this hypothesis were: horizontal "basement" (Rome decollement) , uninterrupted Cambrian-Mississippian sequence to the west, doubled Cambro-Ordovician sequence to the east. The structure fits this simple framework with the additional assumption of subsurface duplication of the Tuscarora-Oriskany interval. Sections cannot be balanced unless the decollement at the Martinsburg level originally continued across the Front; on the west, except where splays that presumably surface in Devonian shale belts are present, there has been no disruption of the stratigraphic sequence. The decollement served as the roof fault for thrusts that ramped from the Rome level and created the associated fold systems. THE VIRGINIA JOURNAL OF SCIENCE 169 EARTHQUAKE TECTONICS IN THE VIRGINIA APPALACHIANS. G. A. Bollinger, Seism. Obs . , Dept, of Geol. Sci . , Va. Polytechnic Inst. & St. Univ. , Blacksburg, Va. 24061. Earthquake activity in the Va. Appalachians varies considerably along strike. In the northeast, from Shenandoah Co. northward, there was a minor amount of activity in the 19th century but very little since then. In their central portion, centered about Bath Co., the Va. Appalachians have been virtually aseismic. The most seismically active portion has been at Giles Co. in the SW part of the state. There, the largest earthquake to have occurred in Va. and the second largest in the SE U.S., took place on May 31, 1897. It was a magnitude 5.8, Intensity VIII shock centered near Pearisburg and felt over some 725,000 km2 This paper- will present results obtained from recent (1978-1983) seismograph network monitoring of the Giles Co. locale. Analysis of the network data has defined a 40 km long, NE-trending zone. That trend puts the zone at a 20-30° disparity with the ENE-trending host southern Appalachians. The zone extends vertically from 5 to 25 km in depth with a dip that is very steep to vertical. The depth range puts the zone entirely in the basement and below the Appalachian decollement. Geological interpretation of these seismic results suggests that the seismogenic fault is probably a reactivated Iapetan fault. PARTIAL EXTRACTION OF METAL FROM MINUS 80 MESH STREAM SEDIMENT AND COATED PEB¬ BLES NEAR AN UNMINED SULFIDE DEPOSIT. T. L. Brown and G. D. Robinson, Dept, of Geology, James Madison Univ., Harrisonburg, VA 22807. Samples of fine-grained stream alluvium and pebbles with Mn-Fe oxide coatings were collected from streams associated with two small sulfide deposits near Andersonville , Va. Three partial extractants, oxalic acid, 10% H2O2 , and EDTA, were used for sample leaching. Each sample type and reagent is evaluated for its anomaly enhancement potential and its ability to delineate the sulfide zones. For both sediment and coatings, the utilization of ratioed data (metal/Mn or metal/Fe) following 10% H2O2 digestion produces the greatest anomaly enhance¬ ment. Analyses of coatings consistently produce greater anomaly contrasts for Zn compared to sediment. Although anomaly contrasts for Cu are similar for both coatings and sediment, the sulfide zones are more precisely delineated by analy¬ sis of coatings. If results of this study are generally applicable in Piedmont exploration, alluvial Mn-Fe oxides constitute a potentially superior sample medium which should be considered for future utilization. The Smith River Allochthon and Thin-Skinned Tectonics in the Virginia Piedmont, by James F. Conley, Virginia Division of Mineral Resources, Charlottesville, Va. The Smith River allochthon is located in the S.W. Piedmont of Virginia and is correlated with rocks of the Inner Piedmont belt of North Carolina. It was thrust over rocks of the Sauratown Mountains anticlinorium and is preserved in a synformal structure between the Sauratown Mountains and Blue Ridge anticlinoria. This synformal structure plunges out both to the southwest in North Carolina and to the northeast in central Virginia. The southeastern edge of the allochthon is terminated against the Chatham fault, the border fault of the Danville Tri- assic basin, and its equivalent to the north. Because, in central Virginia, the southeast limb of the James River synclinorium is composed of rocks of the Smith River allochthon, the structure of this synclinorium is revised. Rocks in the core of the synclinorium are found to be not younger rocks, but older rocks that have been thrust from the east over younger rocks to the west. The structure is not a syncline, but a sequence of imbricate thrust sheets. 170 THE VIRGINIA JOURNAL OF SCIENCE SEISMIC STRATIGRAPHY UNDER THE BLUE RIDGE AND PIEDMONT OF CENTRAL VIRGINIA, Wallace de Witt, Jr.*, and Kenneth C. Bayer*, U.S.G.S., Reston, Va., 22092. A U.S.G.S. common-depth-point reflection seismic profile along Interstate 1-64 from the Valley and Ridge near Staunton, Va. , east to the Atlantic Coastal Plain near Hampton, Va., reveals a regional mega-thrust system whose direction of transport is consistently onshore. Several well-defined seismic-stratigraphic units of regional extent are traceable from outcrops of named and mapped surface formations eastward beneath the Blue Ridge, Piedmont, and the contiguous part of the Coastal Plain. These units form a continuum that is locally cut by imbri¬ cate faults and thickened by subsurface duplication of beds. The combined Lynchburg, Swift Run, and Catoctin Fms. constitute the most extensive seismic- stratigraphic sequence, which extends from outcrops near Charlottesville in the eastern part of the Blue Ridge eastward in the subsurface to depths 50,000- 55,000 feet beneath Richmond. The Chopawamsic Fm. , the Maidens Fm. and Sabot Amphibolite of Glover and others, 1979, make up the 15,000-foot thickness of the eastern Piedmont thrust sheet, which underlies a wide area west of Richmond. Splay faults in the Hylas fault zone appear to cut out these units, and a thick¬ er complex sequence of rocks identified as the Petersburg Granite is exposed at the surface near Richmond or is concealed beneath Coastal Plain sediments. LATE ORDOVICIAN SEDIMENTARY TECTONICS OF THE VALLEY AND RIDGE IN VIRGINIA. Richard J. Diecchio, Dept, of Geology, George Mason Univ., Fairfax, VA 22030. An understanding of the eustatic and isostatic effects on sediment accumulation enables the interpretation of the often more localized tectonic effects. Sea level was probably falling during the early As hg ill, and remained low until the end of the Ordovician. During this low stand, the major portion of the Appalachian Basin was subsiding while sediments of the Queenston clastic wedge were accumulating. Late Ordovician subsidence patterns appear to have been simple in that portion of the basin that is now the Appalachian Plateaus. In the area that is now the Valley and Ridge, subsidence patterns appear to have been more complex. Based on isopach and facies patterns, it can be determined that the Little North Mountain structural front coincides somewhat with each of the following: a boundary between the Reedsville shelf and the Martinsburg basin; the eastern limit of the Orthorhynchul a brachiopod zone; the eastern limit of the Juniata redbeds; and an area of thinning of the Tuscarora Sandstone. It is inferred that the Little North Mountain front represents a tectonic feature that was active at least as early as the Late Ordovician. PRE-TACONIAN DEFORMATION IN THE PIEDMONT OF THE POTOMAC VALLEY — Penobscot ian OR Cadomlan OR BOTH? A. A. Drake, Jr., U.S. Geol. Survey, Reston, VA 22092. Metamorphic rocks in the Potomac Valley comprise five stacked units, from highest to lowest, the Popes Head Formation which unconf ormably overlies all the others, the Piney Branch Complex, the Peters Creek Schist, the Sykesville Formation, and the Eastern Fairfax sequence. These rocks cannot be directly dated, but all are older than the Occoquan Granite that has been dated at about 560 Ma (Pb-U) and 494 Ma (Rb-Sr). Three fold phases have been recognized within the Popes Head. The Occoquan was probably synkinemat ically emplaced during the oldest phase, the Clifton, suggesting Early to Late Cambrian deformation. The Peters Creek and Eastern Fairfax sequence have had a much more complex geologic history. Poly- deformed migmatite and phyllonite olistoliths of Peters Creek Schist are found in the Sykesville Formation, showing that deformation predates the Sykesville, the younger Popes Head Formation, and Clif ton-phase deformation. If the younger age qf the Occoquan Granite is more nearly correct, the Clif ton-phase deformation is probably Penobscotian (Late Cambrian). If the older age is correct, this deformation is probably Cadomian (latest Late Proterozoic - earliest Cambrian). Neither Penobscotian nor Cadomian deformation has been recognized in certifiable North American rocks, so this part of the Piedmont may be a suspect terrane. THE VIRGINIA JOURNAL OF SCIENCE 171 BEDDING PLANE DECOLLEMENTS AND LANDSAT LINEAMENTS: IS THERE A RELATIONSHIP? Thomas M. Gathright , II and Charles B. Stanley, V a. Div. Mineral Resources, Charlottesville, Va. 22903. A correlation between deformation intensity along coal bed decollements and the proximity of LANDSAT lineaments was seen on the Pine Mtn. block in Wise Co., Va. Fracture studies defined two strong, near nor¬ mal, regionally extensive fracture systems of subparallel cleats, joints, cleav¬ age and faults. Pervasive contr actional faults in both fracture systems indi¬ cate both strike-parallel and strike-normal compression in the thrust sheet. The vertical, north trending, left slip Coeburn fault refracts fracture trends and parallels a major lineament suite. Using the Coeburn fault as a model, we propose that lineaments lying at 60 degrees to regional strike, like the Coeburn fault, are vertical zones of flexural slip folding within the systematic joint system. Flexural slip folding about vertical axes should produce differential horizontal displacement, oblique fracturing and deflection of fracture trends in the hanging wall of coal bed decollements beneath the flexural slip zones. OBSERVATIONS OF THE EOCENE DIKE SWARM, HIGHLAND COUNTY, VIRGINIA. Karen J. Gray,*, Dept, of Geol . , George Mason Univ., Fairfax, VA. , 22030, and U.S. Geological Survey, Reston, VA., 22092, and Richard J. Diecchio, Dept, of Geol., George Mason Univ., Fairfax, VA. , 22030. Samples of intrusive rocks, known or presumed to be of Eocene age, have been analyzed by Instrumental Neutron Activa¬ tion Analysis for minor elements. On the basis of these data and previous major-element data, the samples are found to form two main groups: basalts (containing about 50% Si O2 ) and rhyolites (containing about 70% Si O2 ) . The rhyolites are centrally distributed within a larger area of basaltic out¬ crops. This bimodal and spatial distribution is analogous to volcanic systems associated with Cenozoic extensional tectonics in the western United States. The swarm of intrusions trends along the 38th Parallel, and apparently repre¬ sents an eastward extension of the Shawneetown-Rough Creek fault zone. This linear zone extends from eastern Missouri, across southern Illinois, into western Kentucky, and is characteri zed by predominant ly normal faults, igneous centers, and ore deposits. Activity along this fault zone dates from the Precambrian to the Recent. Our new mtnoh-el ement data indicate a rift-related origin of the Highland County dikes. Relative concentrations of stable minor elements suggest minimal contamination by sialic crust. ORIGIN OF THE BALTIMORE MINE MANGANESE DEPOSITS, FORT VALLEY, VIRGINIA. James H. Greene,* Dept, of Geol., George Mason Univ., Fairfax, VA 22030, & Douglas Mose, Dept, of Geol., George Mason Univ., Fairfax, VA 22030. The Ridgely Sandstone on Three Top Mountain, Virginia contains manganese deposits which were mined early in this century. Samples from one of these mines, the "Baltimore Mine", were investigated using optical and chemical methods in an attempt to determine the orogenesis of this deposit. Comparisons are made between the trace element chemistry of this mine and other known deposits. EXPLORATION FOR URANIUM IN PITTSYLVANIA COUNTY, VA. Christopher R. Halladay, Marline Uranium Corp., P.O. Box 1626, Danville, VA. 24543. In July, 1982, Marline announced the discovery of a major uranium deposit 6 miles northeast of Chatham. The orebody contains approximately 30 million pounds of uranium oxide at an average grade in excess of .20% U^O , an<^ larger quantities of lower grade reserves. The extent of the deposit was defined by 256 drill holes totaling 190,000 ft. The orebody lies within a shear zone adjacent to the Chatham Fault, which forms the western boundary of the Danville Triassic basin. The principal host rock is a protomylonitic augen gneiss that was probably derived from gneisses of the Fork Mountain Fm. and from Leatherwood Granite,. The mineral¬ ization is hydrothermal and may be related to an Acadian orogenic event. In searching for this and other deposits. Marline has flown airborne radiometric, magnetic and VLF surveys. Surface exploration methods have included scintil¬ lometer, spectrometer, magnetometer and gravimeter surveys; soil and rock geochemistry; and soil radon gas measurements. 172 THE VIRGINIA JOURNAL OF SCIENCE CONODONTS DOCUMENT CONTINUOUS TO INTERMITTENT DEPOSITION ACROSS THE LOWER-MIDDLE ORDOVICIAN BOUNDARY— NORTHERN VIRGINIA TO BELLEFONT, PA. Anita G. Harris*& John E. Repetski*, U.S. Geol. Survey, U.S. Nat. Mus. E-501, Washington, D.C. 20560. In the Valley and Ridge province, from Lexington, Va., to Tennessee, Middle Ordo¬ vician rocks of Chazyan and (or) early Blackriveran age disconf ormably overlie Beekmantown or Knox Group carbonate rocks that are consistently of latest Early Ordovician age. Physical evidence for this unconformity includes surfaces of considerable relief, solution features, and chert-pebble conglomerates. From Harrisonburg, Va., northward along the Allegheny Front in Pennsylvania, physical evidence for an unconformity between the dominantly Lower Ordovician dolostone sequence and the overlying Middle Ordovician limestones is generally absent. Karstif ication features and scattered chert-pebble lenses, however, occur local¬ ly in the upper several hundred feet of the Beekmantown Group. In this region, conodonts, virtually the only biostratigraphically diagnostic fossils in these rocks, record continuous deposition across the Lower-Middle Ordovician boundary which lies from 900 to 350 feet below the top of the Beekmantown Group. Because the porosity horizon (s) developed at the Lower-Middle Ordovician unconformity is often the target for explorationists, these new age and paleogeographic data significantly affect hydrocarbon and mineral exploration strategies. CHEMICAL WEATHERING OF A HOMOGENEOUS CRYSTALLINE ROCK. Michael Hartley* and Douglas Mose, Department of Geology, George Mason University, Fairfax, VA 22030 The Lynchburg Formation in southwest Virginia is a metasedimentary formation somposed of an upper schist unit and a lower gneiss unit with amphibolite zones located at the base of both units. An amphibolite grade quartz-pl agiocl ase-two mica gneiss of very uniform composition is located at the base of the Lynchburg. The parentage of the QPM gneiss is uncertain, but since a Rb-Sr isochron study of both the Lynchburg and the QPM gneiss indicates that both rocks experienced isotopic homoginization during Paleofoic metamorphism, it is likely that the QPM gneiss was a sedimentary or volcanic rock prior to metamorphism. A study of chemical variations in the QPM gneiss caused by weathering indicates that neither the B soil horizon or the C soil horizon is suitable for age determina¬ tions using the Rb-Sr whole-rock isochron technique. USING COMPUTER PROGRAMS IN GEOLOGY: UNDERGRADUATE EDUCATION. Diana J. Holford* & Peter G. Kimmel, Dept, of Geology, George Mason Univ. , Fairfax, Va. 22030. Computer programs useful to students and teachers of undergraduate geology fall into five basic categories: performing long, repetitive, or complex calcula¬ tions; storing and retrieving large amounts of data; presenting data in graphic (chart) form; simulating real-life events or activities; computer assisted instruction. Programs written in languages like BASIC, FORTRAN, and Pascal are listed in various indexes and periodicals; but because there are few educa¬ tional geology programs available, you may have to write your own. Some sug¬ gestions for writing your own programs: choose the correct language for the type of program you are writing (e.g. BASIC is a good language for most appli¬ cations, but not for data base programs); select a large enough (or small enough) computer to deal with your program; make your program easy to use by providing adequate documentation (manuals, comment or remark lines, and prompt lines) . PERVASIVE CATACLASTIC TEXTURES IN A FELSIC GNEISS UNIT WEST OF THE RICHMOND BASIN, VIRGINIA. Stephen G. Johnson*, Dept, of Geology, College of William and Mary, Williamsburg, Va. 23185. Structural and petrographic studies of the Mosely Felsic Gneiss of Reilly (1980) adjacent to the western border of the Richmond Basin show that the unit is heterogeneous in composition and has cataclastic textures throughout. The Mosely Felsic Gneiss, as mapped, has three primary rock types: felsic gneiss, amphibole gneiss and biotite gneiss. Felsic gneiss is dominant and has the best preserved cataclastic textures, ranging from ultramy Ionite to microbreccia. Three periods of deformation have been identified in the felsic gneiss. This unit may be correlative to the Hylas zone, a well defined cataclastic zone on strike to the north of the Mosely Felsic Gneiss. THE VIRGINIA JOURNAL OF SCIENCE 173 DINOFLAGELLATE PALEOECOLOGY OF THE VIRGINIA COASTAL PLAINS, by Debby Kay* Dept, of Geology, George Mason Uni v. , Fairfax, VA. 22032. Dinoflagellate assemblages from U.S.G.S. core samples in King George County, Virginia have been examined for possible pa leoeco logic indications of the Pal eocene and early Eocene Coastal Plain Province. Formations sampled include the Aquia (Paleocene) and the Nanjemoy (early Eocene) and the Marlboro Clay which is sometimes present between the Aquia and the Nanjemoy. Paleoecologic interpre¬ tations are based on species dominance and diversity patterns. Initial com¬ parisons show a correlation between the appearance of certain species and changes in the relative dominance and diversity ratios which suggest that environmental factors are primarily responsible for the cyst assemblages of the study section. Characteristic species of higher diversity suggest a more offshore environment; lower diversity figures appear to represent a near shore envi ronment. FOSSIL BONY FISH OF THE CHESAPEAKE GROUP, VIRGINIA AND MARYLAND. Peter G. Kimmel , Geology Dept., George Mason Uni v. , Fairfax, V a. 22030. Although tele- osts have been known from the Chesapeake Group since the late 1700 1 s, no comp¬ rehensive survey has been published. A preliminary list of specimens from the Smithsonian Natural History Museum, the Calvert Marine Museum, and private collections show that formations within the group contain: a sturgeon (Acipen- ser) ; a sea catfish (Arius) ; a bonefish (Paralbula) ; a wrasse ( Protautoga) ; several mackerels (Acanthocymbium, Neocymbium, Cymbium); two swordfish (1st i o - phorus, Xiphas) ; a cod (Gadus); a porcupine fish (Diodon); an ocean sunfish (Mol aj; a black drum (Pogonias); a whiting (Menticirrhus) ; a tilefish (Lopho- 1 at i 1 u s ) . The more complete articulated specimens appear to be from offshore muds, below wave base. Other fossil s are isolated, abraded fragments from con¬ glomerates or shell layers and suggest more shallow water deposition. PENETRATION RESISTANCE OF THREE VIRGINIA SOILS. Stephen F. Maher* , Dept, of Geol . , James Madison Univ., Harrisonburg, Va . 22807. Penetration resistance is commonly used to indicate bearing strength in compacted soil masses. Most engi¬ neering specifications require that soil foundations in the field be compacted to maximum density at optimum moisture content. The purpose of this study was to measure penetration resistance of three common Virginia soils representing extremes in texture. Each soil was compacted at a range of moisture contents. Test results indicate that the penetration resistance of the three soils is greater on the dry side of the optimum moisture and maximum density and declines as moisture increases. It is concluded from this study that the maximum pene¬ tration resistance of many soils is reached when the compacted soil is on the dry side of optimum moisture and at lower than maximum density. Since virtually all construction specifications require soil foundations to be placed at optimum moisture and maximum density this study would indicate that such placement may not be at maximum bearing strength. PREDICTING GEOLOGIC HAZARDS IN ADVANCE OF MINING. Marshall S. Miller*, Geolog¬ ical Consulting Services, Bluef ield, Virginia 24605. In the Appalachians, thick coal reserves with overall good mining conditions are becoming more scarce each year. With erratic and decreasing market conditions, the ability to define low-cost high-productivity reserves among the remaining complex coal measures is now a critical requirement for Appalachian coal producers. Predicting geological hazards prior to mining offers the industry potential success for achieving low-cost mining. Among the applications are: depositional modeling; roof and floor rock mapping utilizing drill test data and underground reconnaissance; seismic surveys, geophysical logging; and remote sensing. Classification of lineaments in coordination with over¬ burden and roof rock mapping appears to offer the most useful and productive geological tools for defining potential hazards in advance of mining. Case histories will be demonstrated for mining operations in Maryland, Virginia, West Virginia, and Kentucky. 174 THE VIRGINIA JOURNAL OF SCIENCE 87Sr/86Sr EVIDENCE FOR PAN-AFRICAN TYPE GRANITES IN THE BLUE RIDGE. Douglas Mose, Department of Geology, George Mason University, Fairfax, VA 22030 The Pan-African orogenic belts in Africa are similar to the latest Precambrian geology of the central and southern Appalachians in that both appear to have involved the development of an ensialic basin in latest Precambrian time. In the Appalachians, the basin development is recorded by the Lynchburg Formation and similar sedimentary-volcanic units. Geochemical evidence indicates that this basin was initiated by rifting and the emplacement of peralkaline plutons about 690 m.y. ago. Although the Lynchburg and related units cannot be dated (no fossils; Paleozoic metamorphism has "reset" the isotopic chronometers), the closure of this latest Precambrian basin is marked by the emplacement of per- aluminous plutons about 600 m.y. ago. It appears that this depositional event which involved the Lynchburg and similar units preceded the opening of the Proto-Atlantic by a time interval similar to the interval between deposition of Appalachian graben sediments and volcanics of Triassicage and the opening of the modern Atlantic ocean. Rb-Sr AGE IS ORDOVICIAN FOR THE CAT0CTIN METABASALT AND METARHYOLITE. Susan Nagel* and Douglas Mose, Department of Geology, George Mason University, Fairfax, Virginia, 22030. An 800 million year old age for the Catoctin and related formations (Rankin and others, 1969) is generated by discordant zircons and can not be correct, since Catoctin-type feeder dikes are found cutting the Robertson River pluton, and the pluton has an age of 570 ± 15 million years. A Rb-Sr whole rock study shows that the Catoctin Metabasalts yield an isochron age of 470 ± 70 million years (initial ratio 87sr/g65r = 0.7039 ± 0.0002), and the Catoctin Metarhyolites yield an age of 420 ± 4 million years (initial ratio = 0.7118 ± 0.0099). A consideration of the ages, initial ratios and radiogenic growth of ^Sr suggests that the Catoctin was metamorphosed about 420 million years ago, and no more than 70 million years could have elapsed between the extrusion and metamorphism of the Catoctin. If this interpretation of the Rb-Sr data is correct, than the Catoctin was extruded no more than about 490 million years age, and therefore is probably an early Ordovician unit. GEOCHEMICAL DISCRIMINATION BETWEEN THE NORTON AND WISE COALS, SOUTHWEST VIRGINIA Jack E. Nolde*, Virginia Div. of Mineral Resources, Charlottesville, Va., 22903 Chemical analyses of coal samples from the Norton and Wise formations provide a data base for discrimination. Using analysis of the volatile matter, fixed carbon, ash, total sulfur, iron oxide, titanium oxide and 11 trace elements, discriminant functions were calculated, and used to classify coals. Using analyses of Mn, U, B, Fe203 , Cu, in 38 coals, with 19 from above the Kennedy coal and 19 below the Kennedy coal gives a discriminant function with 84.2% probability of classifying a coal as upper or lower Norton. Analyses of VM, S, Ce, Co, Fe203, Ti02 , U, and V on 34 coals in each group gives a discriminant function with 82.3% probability as being a Norton or Wise coal. The proximate and major and trace elements show that the upper Norton coals have a higher total S, B, Ce, Cu, Fe203, Ni and V, the lower Norton coals containing a higher Mn concentration. The Norton coals average higher in ash and Mn; the Wise coals are higher in B, Ce, Co, Ge, Fe203, Ni and V. THE VIRGINIA JOURNAL OF SCIENCE 175 NUMERICAL ANALYSIS OF GAMMA-RAY LOGS OF THE HINTON FORMATION, SW VIRGINIA Jack E. Nolde*, Virginia Div. of Mineral Resources, Charlottesville, Va. , 22903 The Hinton Formation is a wedge of near-shore marine strata consisting of alternating siltstone, sandstone, and limestone beds in southwestern Virginia. Fourier analysis of 31 gamma-ray logs of the Hinton indicate that there are significant differences in radioactivity among the wells. The gamma-ray logs were digitized, and Fourier coefficients computed, assuming a fundamental period of 24, for the first twelve harmonics. The coefficients were used to compute the discrete power spectrum square root (Cn) , and then subjected to the Kruskal-Wallis test and discriminant analysis. Discriminant analysis on the 12 Cn values yielded six harmonics as being significant difference indicators. The significance was tested with WILK’S lambda and Chi-Square statistic and indicates that the discriminant separates the Hinton Formation into two distinct groups. Use of the discriminant will result in correct group assignment approximately 90% of the time. ENVIRONMENTAL GEOLOGY OF FOUR MILE RUN J.V. O' CONNOR, Dept, of Earth and Life Sciences, Univ. of D.C., Washington, D.C. 20008. A small Fall Zone drainage basin in Northern Virginia, once known for its flood impact, has numerous natural history events and geological processes to observe and investigate over a short linear distance. This small basin (20 sq. miles) provides an ideal case study illustrating a classic contrast between Piedmont and Coastal Plain land uses and associated hazards accumulating downstream. Among the measureable landscape porcesses are fall retreat, spring flow, run-off, tide cycles, terrace origin, slides and erosion. Research potential lies in mineral resources, metamorphism, structure, sedimentation, paleontology, agronomy and climatology. Social history still survives with Indian artifacts, mill-farm sites, mineral springs, road and rail right-of-way. A thirty million dollar flood control project, managed by a computer model, has increased property values, displaced the poor, but stopped the flooding. The tidal mouth supports an airport, marina, railyards, major highways, treatment plant, state boundary and parkland. This watershed is a living short course. GEOLOGY OF AN ALASKITE DIKE ( ?) EXPOSED SOUTHEAST OF CHARLOTTESVILLE, VIRGINIA. Robert A. Russ In * and W. Cullen Sherwood, Dept, of Geol., James Madison Univ., Harrisonburg, Va. 22807. On the east flank of Monticello Mountain southeast of Charlottesville a felsic dike-like rock body crops out within the Catoctin Green¬ stone belt. In hand specimen the rock exhibits fine granular texture and rich pink color. Petrographic analysis shows a predominance of equigranular quartz (51.4%) and perthitic feldspar (34.7%) with minor magnetite (2.2%). Mineralogy and texture conform to the classical definition of alaskite. Field investiga¬ tions indicate the rock body is tabular in shape, and trends parallel to the strike of the enclosing greenstone. While the origin of the rock is obscure the following evidence favors an igneous rather than a sedimentary origin: 1) other metasediments within the Catoctin are quartzites with obvious grain rounding and sedimentary structures, 2) sources for perthic feldspar sediments are not found in the vicinity, 3) the level of metamorphism required to alter a sediment to alaskite would probably have metamorphosed the enclosing greenstone beyond the present greenschist facies, and 4) while no sedimentary structures were found in the alaskite, possible flow structures were observed. 176 THE VIRGINIA JOURNAL OF SCIENCE BROKEN-FORMATIONS OF THE PULASKI THRUST SHEET, NEAR PULASKI, VIRGINIA. Art Schultz*, Virginia Div. Mineral Res., Charlottesville, Va. 22903 Broken-formations (Hsu, 1974; Harris and Milici,1977) occurring in the lower part of the Pulaski thrust sheet contain some of the most strongly deformed sedimentary rocks in the Valley and Ridge province of the Southern Appalachians. Deformation in this zone ranges from grain scale cataclasis to regional scale faulting. The broken-formations are distinguished from rocks structurally higher on the sheet and from rocks of the underlying Saltville sheet by: 1) a sharp increase in the variability of fold and fault styles; 2) greater ranges in fold plunges and dips of axial surfaces; 3) a low degree of preferred orientation of folds and faults; 4) an increase in the frequency of mesoscopic structures and 5) the presence of Max Meadows tectonic breccia. Structural analyses seem to indicate that deformation in the broken-formations is Alleghanian in age and that they formed under elastico-f rictional conditions, possibly under elevated fluid pressures with temporally variant stresses and that lithology may have played an important role in locallizing the broken-formations along the base of the Pulaski sheet. CORONAS IN THE MONTPELIER ANORTHOSITE, HANOVER CO., VA. Ellen R. Stofan*, Dept, of Geol., Col. Wm. & Mary, Williamsburg, Va. 23185. Two types of coronas were found in the Montpelier anorthosite*- located in Hanover Co., Virginia. One type of corona formed around pyroxene, the other type around ilmenite or rutile. Co¬ ronas are common in anorthosites and related rocks. The structures preserve past chemical reactions in the rocks. The pyroxene coronas typically have four shells. The pyroxene itself has been replaced by an amphibole and biotite pseudomorph. Rimming the pseudomorph is a shell of granoblastic biotite and quartz, a biotite mat, a garnet shell, and a zone of recrystallized plagioclase. The rutile and ilmenite coronas have rims of sphene, biotite, and possibly garnet. The formation of all of the she¬ lls can be accounted for by combining the minerals present. The reactions to produce the coronas indicate that the deposit has gone through several stages of metamorphism. The coronas at Montpelier were probably produced as the anorthosite was subjected to retrograde granulite facies to amphibolite facies metamorphism, which supports the general theory that coronas are retrograde features (Griffin and Heier,1973). (Supported by Col. of Wm. & Mary minor research grants) ROCK AND MINERAL RESOURCES IN VIRGINIA. Palmer C. Sweet*, Virginia Division of Mineral Resources, Charlottesville, Va. Mineral production in Virginia in 1982 was valued at $1.57 billion, $221 million less than 1981 due primarily to a DOE projected 5.8 million ton decrease in coal production. Rank of mineral commodi¬ ties produced by total sales yere: coal, stone (crushed and dimension), cement, lime, sand and gravel and natural gas. Virginia led the nation in the production of kyanite, and a feldspar marketed as "Virginia Aplite." Other commodities pro¬ duced include clay materials, glass sand, gypsum, iron-oxide pigments, petroleum, by-product sulfur, talc, and vermiculite. Processing plants imported and pre¬ pared calcium aluminate cement, lithium carbonate, magnetite, manganese, mica, perlite, and vanadium pentoxide. Resources that may be important in the future include uranium, base and precious metals, tantalum and beryllium minerals, dia- tomaceous sediments, industrial silica and additional raw materials suitable for filler and extender uses. Potential waste material resources include flyash, by¬ product carbonate, diabase and granite fines, and high alumina (85%+) slag. THE VIRGINIA JOURNAL OF SCIENCE 177 ALLOCHTHONOUS CLASTS OCCURRING IN THE CHOWAN RIVER FORMATION (UPPER PLIOCENE) OF SOUTHEASTERN VIRGINIA. Andrea E. Victor *, Dept, of Geology, College of Willaim and Mary, Wi 1 1 i amsburg , VA 23185. Allochthonous clasts of Coastal Plain origin occur within the Chowan River Formation (Yadkin's beds“Upper Pliocene) near Yadkin in southeastern Virginia. Ellipsoidal, discoidal and irregular clasts ranging in size from 1 to 50 cm are distributed sporadically throughout the Chowan River. The mineralogy and internal fabric were were examined with x-ray diffraction, electron microscopy and in thin section. Goethi te ( 9 ) , angular quartz, calcite(?), mica and ilmenite(9) occur in the dense fine-grained matrix of the iron-rich clasts. All clasts exhibit fossate, dimpled or shell impressed oxidized surfaces. Locally derived from weathered Yorktown sediments, the clasts were reworked into the Chowan River Formation during the accompanying marine transgression. The presence of Yorktown derived clasts substantiate the presence of the disconf ormaity between the Yorktown Formation and the Chowan River Formation. THE EFFECT OF ACID RAIN ON THE WEATHERING OF SEDIMENTARY ROCKS INDIGENOUS TO ROCKBRIDGE COUNTY, VIRGINIA. Richard W. Young, Daniel W. Armstrong, Robert T. Grauer Jr. , Michael A. Pleva, Scott S. Prysi, James K. Shillington, Dept, of Chem. , Washington and Lee Univ. , Lexington, VA 24450. The purpose of this research project is to obtain data concerning the acid rainfall in Rockbridge County and to apply this data to understand how acid rainfall may accelerate the chemical weathering of limestones, dolomites, and sandstones within the county. Lab results may even be useful in predicting geophysical changes due to acid rain. We devised a simple experiment to measure the chemical weathering of various rocks. Rock samples are collected from lithologically distinct out¬ crops and fractionated into particles of about 2.0 mm in diameter. Synthetic acid rain is made from deionized water by adding sulfuric acid and sodium bicarbonate as necessary to obtain waters of pH 3.0, 4.0, 5.0, 6.0, 7.0, and 8.0. Each of the rock samples is then equilibrated with each of the samples of synthetic acid rain. An Atomic Absorption Spectrometer is then used to deter¬ mine the concentrations of Ca+2 , Mg+2 , and Fe+^ in these solutions. The results should indicate whether or not acid rain speeds the chemical weathering of rocks. Meanwhile, we have collected rain to determine the acidity of rainfall in the Rockbridge County area. Materials Science Section IN-SITU DEPOSITION AND MELTING OF SUBMICRON INDIUM-TIN ALLOY PARTICLES. G. L. Allen*, and W. A. Jesser, Dept, of Materials Science, Univ. of V a., Charlottes¬ ville, V a. 22901 Although the TEM study of the melting point of small, single component, metal particles has been of interest for some time, little is known about particles of binary compositions. For the In-Sn system, particles in the 5 to 50 nm range of diameters were co-vapor deposited onto supported carbon films in vacua of 5 x 10-6 torr to 10"7 torr for in-situ depositions. Evidence has sug¬ gested that the liquidus line of the bulk phase diagram is shifted overall to lower temperatures for particles of high and low In concentrations. In addition, the composition extent of the single phase regions of the phase diagram are dis¬ placed depending on the size of the particles. Dark field microscopy and diffrac¬ tion patterns reveal that amorphous particles are formed for some ranges of com¬ position as determined by X-ray energy dispersive spectroscopy. These nearly co¬ incide with at least two of the three two-phase regions of the bulk phase diagram at room temperature. Particles having compositions which correspond to the sin¬ gle phase regions show normal crystalline structures. In addition, the presence of internal boundaries can be resolved in particles as small as 10 nm in diameter, suggesting the presence of interphase boundaries. 178 THE VIRGINIA JOURNAL OF SCIENCE CAN NUCLEATION THEORY BE APPLIED TO SECOND ORDER TRANSFORMATIONS? R. E. Barker, Jr. , Dept, of Mat. Sci . , Univ. of Va. , Charlottesville, V a. 22901. According to nucleation theory, for a single chemical constituent, local density fluctuations will lead to the formation of a small nucleus of 3-phase within an a-phase matrix if the temperature is less than t£, the normal transition temperature for a t 3. An interfacial free energy will oppose the enlargement of the nucleus and a bulk free energy gV^ will encourage growth, where g = (63 - Ga)/Vg and is the transformed volume. In the standard treatment aG = AH - TAS is approximated, for undercooling 0 = t£ - T, by AH ~ aH^ and AS = AH^/Tf, giving AG ~ ASt • 0. In the present approach g is taken as a power series, g (e ) = a0 + a:0 + a202 + ... . oThermodynamic relations, S = (9G/3T)p, etc., lead to a0 = 0, a1 = -AHt/TtVg, a2 = (ACp/T^-Vg) + 2agAHt/VgT£, ... . This approach not only gives correction terms for first order transformations , it also provides the formalism for extending the theory to higher order transformations. For example, for a second order transition it is predicted that r* ~ 2yV^Tt/ACp • 0, for the crit¬ ical size of a spherical nucleus. X-RAY FLUORESCENCE SPECTROMETRY. Robert A. Bayles, Code 6312 NRL, Washington, DC 20375. X-ray fluorescence spectrometry (XRF) is a technique to perform non-destructive chemical analysis. At the Naval Research Lab XRF is used to help characterize materials to be used in research projects, to identify the material in a component which is the subject of a failure analysis, to re-identify samples having different compositions which were inadvertently mixed up, and to sort scrap. The XRF technique involves exciting electrons within the atoms in a material using a source of x-rays, detecting the x-rays produced as the atoms relax, determining the intensity (number of photons) produced as a funciton of their energy or wavelength, interpreting this spectrum, and, for quantitative results, comparing this spectrum with the spectrum of a standard material of similar composition. The instrument which is discussed is a commercial instrument which, due to thorough engineering, combines high analytical performance and flexibility with convenience and safety. Topics discussed include advantages and disadvantages of XRF, x-ray sources, wavelength and energy dispersion, x-ray detectors, multichannel analyzers, specimen preparation, qualitative analysis, and various approaches to doing quantitative analysis. ANALYZING FRACTURE SURFACES BY PHOTOGRAMMETRY. J. I. Bennetch, Metallurgy Laboratory, Reynolds Metals Company, Richmond, Va . 23261. Photogrammetry , "a science of making reliable measurements by the use of usually aerial photographs in surveying and mapmaking," is also a useful metallurgical tool to characterize microscopic topological features. For instance, in fractography , contour maps accurately portraying three dimensional features of fracture surfaces can be constructed by photogrammetry. In addition, photogrammetry can be employed to correlate certain bulk tensile and fatigue properties with microscopic properties. Fracture surface dimple size, spacing and depth, as determined by photogrammetry, can be related to fracture toughness. Moreover, dimple size is also a function of the stress intensity factor in tensile specimens and the crack growth rate in fatigue specimens . THE GROWTH OF III-V SEMICONDUCTOR MATERIALS BY ORGANOMETALLIC CHEMICAL VAPOR DEPOSITION. Ivan 0. Clark, A. L. Fripp, R. K. Crouch, and W. J. Debnam, NASA Langley Research Center, Hampton, VA, 23665. Vapors from organometallic com¬ pounds such as trimethyl gallium, triethyl indium, and trimethyl bismuth can be used with hydrides such as arsine and phosphine of the group V elements as sources for the growth of epitaxial layers of III-V semiconductor materials. The semiconductor layers produced by this organometallic chemical vapor deposi¬ tion technique have many novel applications in electronics and electro-optics. An overview of ongoing research in this budding field will be presented along with a description of some of the advantages and the unique problems of this technique . THE VIRGINIA JOURNAL OF SCIENCE 179 FRICTION AND WEAR OF AN ION-IMPLANTED Co-RASED ALLOY. S.A. Dillich* and I.L. Singer*, Code 6170, Naval Research Laboratory, Wash., DC 20375. The effects of ion inplantation on the tribological behavior of a centrifugally cast, cobalt-based alloy (50 Co, 31 Cr, 12.5 W, Stoody 3) have been investigated by friction tests and wear scar analysis. The kinetic coefficient of friction, yk, was measured on implanted and nonimplanted Stoody 3 disks using a ball on flat geometry during low speed (0.1 mm/ sec) , dry sliding contact. High friction coefficients (yk-0.6) observed for alloy/ Stoody 3 couples coincided with the formation of debris with the same composition as the softer of the mating alloys in the wear scar. Much lower yk values (yk-0.25) were measured on titanium implanted surfaces. Auger spectroscopy revealed that vacuum carburization of both carbide and matrix phases of the Stoody alloy occurred during titanium implantation. Implantation of the Stoody alloy with nitrogen did not result in low friction during dry sliding tests. THE DUCTILE FRACTURE OF C.P. TITANIUM. Marjorie A. Erickson and H.G.F. Wilsdorf, Dept, of Mat. Sci., The Univ. of Va. , Charlottesville, Va. 22901. The ductile fracture of C.P. titanium was studied, with samples of varying grain sizes, using tensile testing techniques, statistical evaluations, and stereo photo¬ graphic techniques. Statistical studies of grain size, dimple size, intertriple point and interdimple spacings were used to aid in the determination of microvoid Initiation sites. The stereo photographic techniques were used in conjunction with the Scanning Electron Microscope to study microstructural fracture surface characteristics. Stereo techniques such as contour maps and relative height profile lines were utilized in matching dimples on opposing fracture surfaces. These quantitative techniques enabled crack opening modes to be determined and thus allowed for the qualitative evaluation of local stresses and stress state Variations throughout the fracture surface. Knowledge gained of the stress istates that contribute to fracture aid in the study of void growth mechanisms. /Supported by the Office of Naval Research) A TEM STUDY OF THE PEARLITE-RETAINED AUSTENITE INTERFACE. S. A. Hackney* and G. ;0. Shiflet * Dept, of Mat. Sci., Univ. of V a., Charlottesville, Va. 22901. An Fe-C-Mn alloy was studied to ascertain the possible role of crystal lography dur¬ ing the pearl ite reaction. There are two major theories of pearl ite formation and growth in ferrous materials. The first, due to Hull-Mehl, depends heavily on crystallographic effects upon nucleation and growth kinetics of both ferrite and carbide phases. The second, which is generally accepted over that of Hull-Mehl, is due to Hillert. Hillert considers crystal lographic effects as secondary and "fairly unimportant", and relies instead upon the gradual evolution of structure- insensitive, cooperative diffusional growth of the two phases in the formulation of this theory. However, recent demonstrations of partial coherency across ap¬ parently ill-matching fccibcc interphase boundaries by computer simulation and direct TEM evidence has given impetus to the emerging viewpoint that disordered boundaries may be less prolific than previously thought. We will report on our attempts to reanalyze both theories using high resolution transmission electron microscopy as the primary tool. The effect of crystal lography will be noted as well as its relative importance during the pearl ite reaction. 180 THE VIRGINIA JOURNAL OF SCIENCE APPLICATION OF A MAXIMUM-STRESS CRITERION TO CRACK PROPAGATION IN NEUTRON AND He- IRRADIATED STAINLESS STEEL. T. Hanamura* and W. A. Jesser, Dept, of Mat. Sci . , Univ. of V a., Charlottesville, V a. 22901. To better correlate the relationships among neutron and helium irradiation-induced microstructure , tensile test condi¬ tion, and the mechanism of intergranular fracture and flow localization, crack opening mode (Mode I and II) was determined as a new crack propagation parameter and correlated with the local tensile condition near the crack tip by using in- situ HVEM tensile testing of austenitic stainless steel microspecimens. It was found that all grain boundary sliding was observed to be caused by a nearly Mode I crack reaching a grain boundary at an angle near 90° and initiating Mode II cracking at the grain boundary. To explain this behavior, a maximum-stress cri¬ terion was developed. Mode II failure was hindered and Mode I cracking was fa¬ vored by neutron and helium-irradiation, but Mode II cracking could be restored by post-irradiation annealing in the core of helium irradiation. Intergranular failure was observed in the thick regions (25 ym) of helium-irradiated specimens even though the penetration depth of the 80 keV helium was less than 0.5 ym. MICROWAVE ABSORPTION OF FIBERGLASS COMPOSITES. Marvin Hass, Dept, of Physics, George Mason Univ., Fairfax, VA. 22030, & Naval Research Lab., Washington, Dc’ 20375, & T.F. Francavilla, Naval Research Lab., Washington, DC 20375. Fiberglass composites generally consist of glass fibers and organic binders. By studying the temperature dependence of absorption at 10 GHz, it is possible to separate the contributions of each of these components. The "glassy" component is typical of many amorphous materials in being essentially temperature independent in the lighter range and very dependent in the 10-100K region. On the other hand, the "resin" component is temperature dependent near ambient. Its behavior appears quite similar to that of two-phonon difference band absorptions in simple cubic crystals. The results will be discussed in terms of current theories of microwave absorption in glasses and crystals. (Supported by the Office of Naval Research.) STUDIES OF THE EFFECTS OF MOISTURE SORPTION ON ELECTRICAL CONDUCTION IN ION- DOPED POLYMERS. J. A. Hawk*, D. Y. S. Chen*, and R. E. Barker, Jr., Dept, of Mat. Sci., Univ. of Va., Charlottesville, Va. 22901. Previously Barker, Chen, and Huang have examined the process of charge transport in rigid rod polymers such as poly(p-phenylene benzobisthiazole) , or PPBT, by measuring two types of transient currents. The PPBT is an extended chain aromatic hetero-cycl ic polymer which exhibits both a gradual current decay over long periods of time, as well as large jumps in the relative current (i.e., Al/Im-jn) when the PPBT sample is sub¬ jected to increases in relative humidity from essentially dry conditions. Exper¬ iments were conducted on PPBT samples doped with various solutions of LiCt, NaC£, CaCt2> LaC£3, etc. In addition, undoped PPBT fibers were also tested, against which the doped results were compared. In the present work our goal has been to see if the transient current phenomena curve is distinguishably different for different types of polymers, or if there was something unique about the rigid rod polymers. Additional experiments have been performed on fibers of nylon 6,6 and polyethylene (PE) in both the doped and undoped form for a variety of experiment¬ al conditions. The most interesting general result is that each material appear? to have its own characteristic type of Al/Im-jn curve. NDE ENGINEERING. Amos E. Holt, Babcock & Wilcox Company, Research & Development Division, Lynchburg Research Center, P. 0. Box 239, Lynchburg, Va. 24505. This paper discusses the expanding field of Nondestructive Evaluation Engineering (NDE) in industry. This is not currently a separate discipline in academia, however, NDE uses the basic principles developed in classical engineering dis¬ ciplines. NDE provides a mechanism for determining the remaining life of a com¬ ponent, its fitness for use and its ability to perform the desired function without rendering the component useless. THE VIRGINIA JOURNAL OF SCIENCE 18 TRANSIENT DIELECTRIC PROPERTIES OF POLYMER-DILUENT SYSTEMS. C.-C. Huang* and R. E. Barker, Jr., Dept, of Mat. Sci . , Uni v. of Va. , Charlottesville, V a. 22901 Recent work by Chen and Barker has provided persuasive evidence that a useful ex¬ tension of the "weak electrolyte" model (of Barker and Sharbaugh) to the case of anisotropic polymer systems is possible. In continuing our studies of this and other more general features of transport mechanisms in polymers, quantitative in¬ formation was needed for the dielectric permeativity tensor (x-jj) and for its variation with temperature, frequency, and the concentration of various diluents and dopants. The present work relates some of the experimental approaches under¬ taken with these goals in mind and in particular addresses the relevant fact that during diffusion of a diluent into a polymer, the system is inhomogeneous and the effective average dielectric properties (K' , K", or tan 6 = K"/K' ) have a time dependence which is related to the diffusion coefficient. A combination of di¬ electric cells (including a liquid immersion cell) and bridge techniques have been applied to examine PMMA , PS, PTFE, and PPBT (polyparaphenylene benzobisthia- zole), in addition to a quartz standard. A MICROSTRUCTURAL STUDY OF DUCTILE FRACTURE OF AN ALUMINUM COPPER ALLOY. David D. Make!* and H. G. F. Wilsdorf , Dept, of Mat. Sci., Univ. of Va. Charlottes¬ ville, Va. 22901. The ductile fracture of an Aluminum-1 .79w/oCopper alloy has been studied. Large and small polycrystal line tensile samples, heat treated to contain a solid solution, G.P. zones, and e 1 precipitates were strained to frac¬ ture and observed with scanning and transmission electron microscopes. Using stereo microscopy, the fracture surfaces of the bulk samples have been character¬ ized with a special emphasis on analysis of the ductile dimples. This is used to determine the local modes of the crack opening and to gain insight into the nucleation, growth, and coalescence of the voids. Post-fracture TEM and STEM studies of the ligament remnants show the crystallography and general configura¬ tion of the final separation. These results, with related research, have been used to formulate a model of the final separation process. (Supported by the Materials Division of ONR.) TRANSVERSE STRENGTH IMPROVEMENTS IN METAL MATRIX COMPOSITES. A. Y. Teng* and F. E. Wawner, Dept, of Mat. Sci., Univ. of Va., Charlottesville, Va. 22901. Tensile tests of SiC monofilament reinforced aluminum composites show transverse strengths less than 4000 psi. This strength is lower than the matrix strength and is closely related to SiC/A£ bonding limitations. One solution to this prob¬ lem is to make hybrid composites using a smaller and different type of fiber to reinforce the transverse direction without adversely affecting axial properties. The fibers used are the continuous Nicalon and FP fibers with a diameter range of 10-30 ym. Results showed improvements in the composite transverse strengths. However, some problems were encountered which limited performance to that pre¬ dicted by the rule-of-mixture. 182 THE VIRGINIA JOURNAL OF SCIENCE Medical Sciences Section FUSION OF BIOMEMBRANES BY DIVALENT CATIONS, Kenneth Barfield*, David Bevan, Department of Biochemistry, VA Polytechnic Institute, Blacksburg, VA 24061. A number of biological and cellular processes involve the fusion of biomemb¬ ranes, but these fusion events are not well understood. The ability of Ca^+ and Mg2+ to fuse phospholipid vesicles has been documented, but no such work has been done with the divalent cations Zn2+, Cd2+, and Hg2+. Resonance energy transfer between the fluorescent phospholipid analogues N-NBD-phosphatidyl- ethanolamine and N-(lissamine Rhodamine sulfonyl)phosphatidylethanolamine can be used to assay fusion in vesicles containing the labelled phospholipids. In studies with small unilamellar vesicles composed of dipalmitoyl phosphatidyl¬ choline (DPPC) and bovine brain phosphatidylserine (PS) it was found that the divalent cations caused fusion of the vesicles with the relative effectiveness being Zn2+>Cd2+>Ca2+>Mg2+mHg2+. In addition it was found that fusion occurred with as little as 20% PS in the vesicles, which is considerably lower than has been previously reported. ANATOMY AND PSYCHOPATHOLOGY OF LARYNGEAL MUSCLE VOICE VIBRATIONS AND INTERPRETATIONS OF NORMAL AND NON-NORMAL STRESS REACTIONS. John C. Bartone, Department of Research, American Health Research Institute , Ltd . , Annandale, Va. 22003. Normal and non-normal voice vibration stress reactions have not been completely elucidated . In known reactions, as practiced today, emphasis is limited, rigid and dogmatic. Stress reactions produce more than "innocence", fight or flight stress. Psychopathology is greatly inherent in parameters as stress resultant in "freeze", "feint", "feign" or animal acts as "die on the spot", equal to human hysteria, silent or otherwise. Such parameters highly complicate human experimentation. In addi¬ tion ego-status, academic shock, fear of failure, other syndromes lessen validity of such conclusions for science, medicine or law. Inherent psychopathology of stress reactions persistently remains. Psychodiagnostic profiles appear necessary to increase validation. L- LACTATE DEHYDROGENASE IN MITOCHONDRIA? R.B. Brandt, E.S. Kline, S.B. Tinsley* K.S. Rogers, E.S. Higgins, M.G. Waters* and H.R. Seibel, Depts. of Biochemistry and Anatomy, Med. Col. of Va. , VCU, Richmond, Va. 23298. Experiments on cat¬ abolism of D- and L- lactate by us, indicated that some L-lactate oxidation was taking place in the mitochondria. Mitochondrial and cytosolic fractions from rat brain, kidney, heart and liver were assayed for NAD+ dependent L-lactate de¬ hydrogenase (L-IDH) , cytochrome oxidase (cyt ox) and phosphoglucomutase (PGM) . PGM was used as a marker of mitochondrial contamination by cytosol and cyt ox was used to determine contamination of cytosol by mitochondria. The % LDH activity (mito/cytosol) on a vet weight basis was: liver 1.8; kidney 0.8; heart 0.5; and brain 1.2. From activity measurements of PGM in mitochondria there was no contamination by cytosol except in brain preparations. Both by PCM ac¬ tivity and EM, brain mitochondrial samples were contaminated with cytosolic com¬ ponents. Skilleter and Kun (1974) had shown L-IDH in liver mitochondria and we extend this finding to kidney and heart. Isozyme patterns are not the same for all tissues in the cytosolic and mitochondrial fractions. Respiratory control studies showed that oxidation of L-lactate oxidation was coupled to phosphory¬ lation . (Supported by funds from NPCR and Univ. Grant- in-Aid, VCU) . THE VIRGINIA JOURNAL OF SCIENCE 183 CADMIUM METABOLISM BY RAT LIVER SINUSOIDAL CELLS. Thomas J. Caperna* and Mark L. Failla, Dept, of Biochem. and Nutr., V a. Polytechnic Inst, and State Univ., Blacksburg, VA 24061. The metabolism of Cadmium (Cd) was investigated in rat liver sinusoidal cells isolated by collagenase dispersion and purified by cen¬ trifugal elutriation. Initial studies demonstrated that both Kupffer cells (KC) and endothelial cells (EC) accumulated significant amounts of Cd following in¬ jection (S.C.) of the metal salt (15 mg Cd/Kg body weight). When metal lothio- nein (Mt) was assayed in cells isolated from control rats, low levels were pre¬ sent in KC, EC and hepatocytes (HC). Cd injection significantly increased Mt levels in all liver cell types; however, EC contained almost twice as much Mt/mg cell protein as KC and HC. Cd transport and metabolism by KC and EC was studied in vitro using primary monolayer cultures. The results confirmed that EC have i- greater capacity to accumulate Cd than KC. These data indicate that the cells lining the sinuses of liver have a significant role in the metabolism of trace metals and the handling of toxic heavy metals. (Funded by NIH AM 363941). COMPUTER ANALYSIS OF SPECTRAL ABSORBANCES SEPARATING RATS FED 4%, 15%, OR 40% PROTEIN AND STRESSED BY RESTRAINT OR NOISE. Germille Colmano, and R.W. Berlien, Coll. Vet. Med., Div. Vet. Biol. & Clin. Stud., & Gary Nunn, Computing Re¬ sources, VPI&SU, Blacksburg, VA 24061. Rats (21 day old) were equilibrated 1 week and pair-fed on a diet containing 4% casein. Then, the daily intake of 15% and 40% casein fed rats was adjusted so that each animal received the same daily ratio by weight. Immobilization or noise-stress from 36 to 41 days of age was finally added. Data on the level of several protein components of blood plasma, collected before and after stress, and representing only 1/3 of the data availa¬ ble for analysis, are presented. The results indicated that this technique successfully distinguished several different blood plasma proteins, differ¬ entiating across animal groups the 3 different blood protein levels, which corresponded to the 3 levels of casein fed to the rats. The matrix of F ratios for the 3 levels of protein before stress indicated a clear separation of groups. Also, a classification summary and F ratios for the stressed versus the non-stressed animals indicated the possibility of differentiating blood spectra between these groups. DEVELOPMENT OF A RAT MODEL FOR COMPUTER-RECORDED SCIATIC-NERVE SIGNALS (BEFORE INJURY) TO ASSIST REPAIR USING METAL (Na, K, Ca) MONOLAYERS COATED SUTURES. Germille Colmano, and J.M. Gregg, Div. Vet. Biol. & Clin. Stud., Coll. Vet. Med. ; W.E. Schwab, Dept. Biol.; D.J. Schneck, Biomed. Engr. ; W.G. Herbert, Car¬ diac Reabil.; and C.D. Berlien, Anaerobic Micro., VPI&SU, Blacksburg, VA 24061. A trifasicular sciatic nerve segment (15 mm) of rats was exposed at midpoint between spinal chord and knee, and its action potential, from bipolar stimu¬ lating-recording electrodes, was computer recorded. After epineural decom¬ pression, an 8 mm length of nerve was resected from both sides. The counter¬ lateral sides were grafted and allowed to spontaneously degenerate and/or regenerate. The right side was sutured with 9° nylon Ethicon and the left side was sutured with 9° reabsorbable (polygalactine) Vicryl microsutures coated with monomolecular layers of metal (Na, K, Ca) stearate films. Initial record¬ ings of the electromagnetic field potentials on the left and right sciatic nerve, and recordings after two months indicate the feasibility of a computer- coordinated system. 184 THE VIRGINIA JOURNAL OF SCIENCE CHL0RDEC0NE*(CD) EFFECTS ON HEPATIC MICROSOMAL BENZO a PYRENE (BP) METABOLISM L,S. Crouch and R. E. Ebel , Dept. Biochem., Virginia Tech, Blacksburg, VA 24061 CD, a polychlorinated hydrocarbon pesticide, induces cyt P-450. P-450 metabo¬ lizes PAH's such as BP to potent carcinogens and thus possible alterations in BP metabolism as a result of CD exposure are of concern. CD treatment of rats increases P-450 up to 2.7-fold compared to 2.6- and 1.6-fold increases, respec¬ tively, for phenobarbi tal (PB) and 3-methyl cholanthrene (3-MC). BP metabolism measured by 3-hydroxy BP production (AHH) per nmol P-450 was 40, 40 and 270% of control values for CD, PB, and 3-MC microsomes, respectively. In vitro, 7,8- benzoflavone activates AHH in control, CD and PB microsomes but inhibits 3-MC. CD, in vitro, inhibits AHH in control and CD microsomes to a comparable degree based on total CD levels. PB microsomes are less sensitive and 3-MC insensi¬ tive to CD. Total BP metabolism is similar for control and CD microsomes. The data are consistent with similarities in the cyt P-450 pools of hepatic micro¬ somes isolated from control and CD treated rats. STIMULUS/SECRETION COUPLING IN HUMAN LEUCOCYTES: A KINETIC ANALYSIS. Robert W. Dougherty* and Richard J. Freer* Dept, of Pharmacol., Med. Col. of Va. , Richmond, VA 23298. Secretion from DMSO stimulated HL-60 promyelocytic leukemia cells elic¬ ited by CHO-Met-Leu-Phe (fMLP, 10 ~'M) is rapid (ti^<5 sec) and accompanied by en¬ hanced turnover of inositol phospholipids (IP) ana phosphatidic acid (PA). In cells prelabelled with 32p , fMLP (10 'M) elicited rapid (within 5 sec) decreases in radioactivity associated with phosphatidyl inositol and phosphatidyl inositol - 4,5-bis phosphate with simultaneous increases in the labelling of PA and phospha¬ tidyl inosi to! -4-phosphate. Binding of fMLP had not reached steady-state condi¬ tions by 30 sec-a time when secretion and relevent lipid changes are complete- suggesting that only partial occupancy of receptors is sufficient to elicit max¬ imal secretion. Based on the appropriate kinetics and agonist concentration de¬ pendency of both parameters measured, we conclude that IP and PA turnover may be key links in events coupling receptor occupancy and secretion. (Supported by the MCV Periodontal Research Ctr. and the Cystic Fibrosis Fndn.) CHANGES IN SPEED-RELATED PROPERTIES OF SINGLE MOTOR UNITS IN THE CAT PRODUCED BY FATIGUE. L. Dubose* and H.P. Clamann* ( Sponsor : J.L. Poland) Dept, of Physiol, and Biophys., Med. Col. of Va., Richmond, Va. 23298. Motor units typically dis¬ charge at rates (30-60 per sec.) well below those required to produce fused teta- ni and maximum tension; yet in man maximum contractions of muscles are readily produced. To examine this paradox, single motor units of cat gastrocnemius were studied. Twitch contraction time, half-relaxation times of twitches and of un¬ fused tetani, and rate of rise of tetanic tension were measured at the onset and at later times during repeated phasic contractions. Phasic contractions were produced by trains of 13 stimuli at 40/sec. repeated every second. Fast motor units slowed progressively as shown by all above measurements; slow motor units increased their speed. We conclude that the dynamic properties of motor units change during contraction to reduce the differences between the fastest and slow¬ est. Fast units can slow enough to produce maximal fused tetani when driven at 40-60 pulses/sec. (Supported by an N.S.F. Fellowship to L.D.) SYNTHESIS AND- CHROMATOGRAPHY OF PYROGALLOL AND ITS SULFATE ESTERS. D. M. Dulik* and W. H. Soine, Dept, of Pharm. Chem. , MCV/VCU, Richmond, VA 23298. Pyrogal- lol ( 1 , 2, 3-benzenetriol, PY) is a polyphenol of both industrial and pharmacolog¬ ical importance. Studies on its metabolic fate have been limited. Sulfate es¬ ters of PY and 3-methoxycatechol (3MC) were prepared by reaction with pyridine- sulfur trioxide in pyridine at -20°C. Optimal reversed-phase HPLC separation of these sulfate esters was achieved by addition of an ion-pairing agent (di-n-octy- lamine or tetra-n-butylammonium hydroxide) into the mobile phase. Retention be¬ havior was found to be highly dependent upon both the type and concentration of pairing agent. Determination of the presence of PY and 3MC sulfate esters as urinary metabolites was accomplished by ion-pairing HPLC. %-PY was adminis¬ tered to a rat as a single dose (25 mg/kg, ip); 75% of the radioactivity was ex¬ creted in the urine after 12 hours. HPLC separation of urine samples showed the presence of PY mono- and disulfate esters, 3MC sulfate ester, and two other more nolar metabolites: no free PY was detected. THE VIRGINIA JOURNAL OF SCIENCE 185 THE BROAD-HOST RANGE STREPTOCOCCAL PLASMID pIPSOI : IDENTIFICATION OF PLASMID- ENCODED PROTEINS. R.P. Evans, Jr. and F.L. Macrina. Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond. The conjugative streptococcal resistance plasmid pIP50I (30 kb) encodes resistance to chloramphenicol and erythromycin (Emr). Using recombinant DNA methodologies, sequences from a cryptic multicopy plasmid, pVA380-1 , were sub¬ stituted for the p I P501 Emr determinant in vitro. The resulting recombinant plasmid which was recovered in transformed S. sanguis cells and designated pVA797 retained conjugal transfer ability (Tra+). Conjugal transfer deficient (Tra-) deletion derivatives of pVA797 were obtained in vivo. The entire pVA797 molecule and Tra" deletion derivatives were separately cloned into the E_. coli plasmid vector p0P203(A2+). These chimeric plasmids segregated into minicells where they directed protein synthesis. Tra“ derivatives of pVA797 were missing some polypeptides encoded by pVA797. NAEGLERIA FOWLERI CYTOPATHOGENESIS FOR RAT NEUROBLASTOMA CELLS . D. E. Fulford and F. Marciano-Cabral . Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, Va. 23298. Naegleria fowleri nN68 (ATCC- 3089^), maintained in axenic culture for over 10 years, demonstrates a cyto¬ pathic effect for rat neuroblastoma (B-103) cells. Cytopathogenicity is measured by the specific release of 51Cr from radiolabelled B-103 cells. The release of radiolabel is correlated with, but is more sensitive than scoring cytopathogenicity by light microscopic examination of target cell cultures. Live amoebae and amoebic lysates induce a range of specific release of radio¬ label of 10 to 80% from 51Cr labelled neuroblastoma cells depending on the amoebae to target cell ratio or lysate concentration. Heating the lysate re¬ duces cytopathic activity. Incubation of the lysate for 10 minutes at 100 C, 30 minutes at 60 C, or 1 hour at 44 C substantially reduces the cytopathic activity. The cytopathic effect of the lysate is reduced by incubation at 37 C for 2 6 hours, however, incubation at 25, 4, or -20 C for 26 hours has no effect on the lysates activity. The use of the 51Cr release assay and amoebic lysates will be useful in the investigation of Naegleria induced cytopathogenicity. (Supported by Virginia Electric and Power Company and NCI training grant 5 T32 CA 09210-05)* ACCUMULATION AND SUBCELLULAR DISTRIBUTION OF COPPER IN THE RENAL CORTEX OF STZ- DIABETIC RATS. Carol A. Gassman* and Mark L. Failla, Dept, of Biochem. and Nutr., Va. Polytechnic Inst., Blacksburg, VA 24061. The concentration of copper in kidneys from STZ-diabetic rats increased up to 8.7 times that of con¬ trols when animals were fed purified diets (15-20 ppm Cu) ad libitum for 4 weeks. The additional complement of copper was localized entirely in renal cortex. All of the excessive copper present in the cytosol fraction (165,000 x g superna¬ tant) was bound to metal lothionei n. Separation of organelles from the particu¬ late fraction (165,000 x g pellet) by differential centrifugation revealed that the lysosomal fraction from 3 week STZ-cortex contained 16-fold more copper than that from controls. The presence of significantly increased quantities of copper in lysosomes from diabetic rats has been confirmed by transmission electron microscopy. (Supported by Jeffress Trust Fund and Pratt Animal Nutrition Program) . 186 THE VIRGINIA JOURNAL OF SCIENCE PRESSOR DOSE OF ANGIOTENSIN (A-II) vs. PRESSOR DOSE OF A-II + UREA Jeffery Gipson, Dept, of Chem. , Va. Union Univ., Richmond, VA 23220, and Karl C. Corley, Dept, of Physiol., Med. Col. of Va., Richmond, Va 23298. The jugular vein and carotid artery in New Zealand white rabbits (anesthetized by 30 mg/kg Na pentabarbi tol i.p. or i.v.) were cannulated for infusion of test substances and blood pressure measurements, respectively. Alternate infusion (0.5 ml/min) of D5W and increasing doses of A-II in D5W until a pressor effect (+20 mm Hg diastolic) was observed (125 ± 12 ng/kg/min; mean ± SE; N = 9). The pressor dose of A-II in 6.6 M urea was then determined to be 30 ± 5 ng/kg/min. The pressor dose was, therefore, significantly reduced by urea (t = 11.82; df = 8; p < 0.001). Adjustment of the pH of the pressor dose of A-II + urea from 7.8 to 7.0 appeared not to affect the potentiation. Infusion of equiva¬ lent quantities of A-II and 6.6 M urea (0.2 ml/min) into different veins was done in an attempt to determine if the potentiation depended on a reaction between the two substances before infusion. The potentiation was still ob¬ served. Therefore, the enhancement of A-II pressor effects by urea does not appear to be related to the effects of pH or the formation of a new species. (Supported in part by NIH HL07309) SYNTHESIS AND BEHAVIORAL COMPARISON OF C ONF ORMAT IONALL Y RESTRICTED AMPHETAMINE ANALOGS. A. E, Hauck*, R. Young* and R. A. Glennon*. Department of Pharmaceu¬ tical Chemistry, MCV/VCU, Richmond, VA 23298. Rigid analogs of known medicinal agents often demonstrate enhanced activity and/or facilitate our understanding of conformational requirements for efficient receptor binding. In an attempt to determine the active conformation of amphetamine, several conformationally re¬ stricted analogs were evaluated. 6-Amino-6 , 7 , 8, 9-tetrahydro-5H-benzocyclohep- tene was prepared by the method of Cannon, et al and via a thallium trinitrate ring expansion reaction. 7-Amino-6, 7,8, 9-tetrahydro-5H-benzocycloheptene was synthesized by a modification of the procedure of Anand. These and related ana¬ logs, as well as amphetamine and its isomers, were evaluated and compared in a drug discrimination paradigm using rats trained to discriminate (+)- amphetamine from saline. None of the conformationally restricted analogs was as active as (+)-amphetamine. (Supported in part by funding from the A. D. Williams Fund). EFFECTS OF STIMULATION OF THE MESENCEPHALIC DORSAL RAPHE NUCLEUS ON PLASMA LH AND PROLACTIN IN ESTROGEN-PRIMED AND N0N-PRIMED 0VARIECT0MIZED RATS. Connie S. Ki tts* and James H. Johnson. Dept, of Anatomy, MCV/VCU, Richmond, VA 23298. Stimulation of the dorsal raphe nucleus (DRN) in ovariectomized (0VX) rats increases plasma prolactin (PRL) and decreases LH. Serotonin (5HT) mediates the inhibition of LH release (Arendash, Endocrinol. 102:1199). Estradiol benzoate (EB) priming of 0VX rats reversed the effect on LH of stimulation of the basal hypothalamus from inhibition to stimulation (Endocrinol. 99:659). The present studies were undertaken to learn whether a similar reversal could be observed with stimulation of DRN. Rats 0VX at least 4 weeks were implanted with bipolar electrodes in DRN and a cannula in the external jugular vein. Blood samples collected from these unanesthetized rats were assayed by RIA for LH and PRL, and means observed before and during stimulation were compared statistically. Stimulation of DRN decreased LH (460-310 ng/ml ; p<. 006), and increased PRL (32-112 ng/ml; pc. 002). In rats primed with EB (50yg/kg, 2 days) stimulation of DRN increased both LH (126-173 ng/ml; p<.03) and PRL (247-617 ng/ml; pc. 05). We conclude that EB alters the mechanisms by which 5HT neurons in DRN influence the activity of LH-RH producing neurons in the basal hypothalamus. ( Supported by NIH Grant HD12165). THE VIRGINIA JOURNAL OF SCIENCE 187 THE INFLUENCE OF ETHANOL AND CORTICOSTERONE ON REPRODUCTIVE FUNCTION IN THE ADULT FEMALE RAT. R.J. Krieg Jr. , K.A. McGregor*, and L.N. Murad*. Dept, of Anatomy, MCV-VCU, Richmond, VA. 23298. These studies were designed to investigate the effects of ethanol (EtOH) , and corticosterone (CS) on estrous cyclicity, on the proestrous surges of lu¬ teinizing hormone (LH) and prolactin (PRL), and on PRL receptors in the liver. Adult female Sprague-Dawley rats received drinking solutions consisting of tap water, or 4% EtOH in tap water, or one of two doses of CS (40ug/ml , or 400 ug/ml ) in 4% EtOH, for a period of 6 weeks. Although comparable doses of CS had been shown to delay the onset of puberty in young animals, neither dose of CS caused a disruption of estrous cyclicity in adult animals. EtOH caused no change in cyclicity. The preovulatory surge of LH was not affected by EtOH or CS, and ovulation was readily observed in all four groups. The proestrous surge of PRL was not affected by any of the treatments. There was a significant affect of EtOH and CS on liver PRL receptors, however. EtOH and low-dose CS in 4% EtOH decreased PRL receptors by 25.3% and 20.0%, respectively; and high-dose CS ; decreased PRL receptors by 43.8%. These results may be significant with refer- | ence to the relatively common occurrence of gynecomastia in adult human alcoholics. (Supported by NSF Grant #PCM7 9-22155). FIBRINOGEN WHITE MARSH-A NEW HUMAN FIBRINOGEN VARIANT WITH A-a CHAIN DEFECT. J. G. Magnant, R. M. Vennart, H. J. Evans, J. A. Koehn, G. D. Qureshi, Dept, of j Med., Med. Col. of Va. , Richmond, Va. 23298. Congenital dysf ibrinogenemia is a ] rare disorder which may lead to a compromised hemostatic function. A 19 year old female bled excessively following childbirth. Her past and family history | for bleeding tendencies or thrombosis was negative. Her prothrombin time was 14 sec. (control 12), activated partial thromboplastin time 34 sec. (control 25) j thrombin time 44 sec. (control 19), reptilase time 34 sec. (control 25). Her I fibrinogen was 79 mg/ dl by a clotting time method and 384 mg/ dl by immunological ■ method. Similar abnormalities were found in her mother and three brothers. The l kinetics of cleavage of her purified fibrinogen with thrombin, reptilase and southern copperhead snake venom demonstrated markedly impaired f ibrinopeptide A release and normal release of f ibrinopeptide B. The rate of fibrin monomer polymerization was normal. The amino acid sequence studies of A-a chain of her purified fibrinogen demonstrated Argp0->His substitution which appeared to be responsible for the abnormal behavior of her fibrinogen. This fibrinogen variant is designated as, "Fibrinogen White Marsh" after the patient's place of origin in Virginia. EFFECTS OF PARTICULATES ON THE UPTAKE AND METABOLISM OF BENZ0(A)PYRENE . Walter E, Manger*, David R. Bevan, Dept, of Biochem., VA Polytechnic Inst., Blacksburg, VA 24061. The carcinogenic potency of benzo(a)pyrene (BP), a polynuclear aro¬ matic hydrocarbon, is known to increase when instilled or inhaled with particu¬ late matter. Mechanisms for this phenomenon are not known. In the present study, the effect of two particles, one fibrous (Canadian chrysotile) and the other non-fibrous (hematite), on BP metabolism as a function of uptake rate into liver microsomes isolated from 3-methylcholanthrene-treated animals was observed. In addition, the effects of the particle on enzymes involved in BP metabolism, i.e., cytochrome P-450, and epoxide hydrase, have been studied. The metabolism of BP is increased when it is taken into the microsomes from the particle-adsorbed state as compared to the microcrystalline state as would be expected based on the total amount of BP available to the microsomes, even though the particles inhibit the activities of the two enzymes mentioned above. THE VIRGINIA JOURNAL OF SCIENCE 188 CHLORDECONE (CD) POTENTIATION OF CCU HEPATOTOXICITY IN RATS AND GERBILS. E.A. McGrath* and R.E. Ebel , Dept. Biochem., Virginia Tech, Blacksburg, VA 24061 CD and phenobarbi tal (PB), but not mi rex, treatment have been shown to poten¬ tiate the hepatotoxicity of CCI4 in rats. Each of these compounds induces cyt P-450, the enzyme responsible for the production of toxic metabolites of CCI4. CCI4 hepatotoxicity is monitored by increases in SGPT and SGOT, decreases in cyt P-450, histopathological changes, and a dramatic loss of a 50 kdal protein (SDS-PAGE). These changes are a function of CCI4 dose. The gerbil was chosen as a model for human CD exposure since the metabolism of CD is similar in these species. As with the rat, CD, PB and mirex are inducers of cyt P-450. How¬ ever, the uninduced gerbil is significantly more sensitive to CCI4 than the rat. Treatment of male gerbils with CD, PB or mirex has a protective effect compared to control animals with respect to CCI4 toxicity. Of the three, CD provides the least protection while the effect with PB or mirex is comparable. STUDIES ON SOME POTENTIAL SEROTONERGIC AGENTS. J. D. McKenney*, J. M. Jacyno*, and R. A. Glennon*, Department of Pharmaceutical Chemistry, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia 23298. A novel series of N,N-dimethylisotryptamine (isoDMT) derivatives, i.e. derivatives of l-(2-diraethylaminoethyl) indole, was prepared and found to be isosteric with their corresponding N,N-dimethyltryptamine (DMT) counterparts with respect to serotonin receptor (rat fundus) affinity. Whereas the isoDMT derivatives pos¬ sessed a greater affinity than did their corresponding DMT derivatives, they were relatively ineffective in displacing [%]-5-HT binding from rat brain (cortex) homogenates. In a drug discrimination paradigm, using rats as sub¬ jects, 6-OMe isoDMT produced effects similar to those of 5-OMe DMT. (Supported in part by USPHS grant DA-01642 and research funds from the A. D. Williams Fund.) ONSAGER'S RECIPROCITY IN REACTION-DIFFUSION SYSTEMS: A NETWORK THERMODYNAMIC APPROACH. Donald C. Mikulecky, Dept, of Physiol. & Biophys., MCV/VCU. The King-Altman diagramatic method for deriving kinetic expressions for enzy¬ matic reactions has been extended to reaction-diffusion systems (especially active transport) by T. Hill (Free Energy in Biology, Academic Press, NY, 1977). Peusner has developed an alternative, but complementary network thermodynamic representation (J. Chem. Phys. 77: 5500-5507, 1982), which demonstrates that new flow-force coordinates are necessary to exhibit Onsager's reciprocity in these systems. The new coordinates are based on finding a potential which is compatible with the metric used to "measure" the irreversible dissipation in the system. In this work, Mason's unistor is introduced as a means to represent nonreciprocal systems and it is shown that the principle of detailed balance insures that these nonlinear systems are reciprocal at equilibrium and obey Onsager's reciprocity at that point. PROTEOLYTIC ACTIVITY OF GUINEA PIG SPERM. Noel 0. Owers, Department of Anatomy, Virginia Commonwealth University, Richmond, V a. 23298. The aqueous extracts of epididymal sperm digest a fixed gelatin membrane (Owers and Blandau, Chapter 11, in "Biology of the Blastocyst, Univ of Chicago Press 1971), forming circles whose area is proportional to concentration. In¬ dividual sperm incubated on the membrane also digest it forming areas of lysis associated with the acrosome. It has not been known whether the proteases in the acrosome undergo changes during passage of sperm through the uterus, vagina and oviduct, or not. Female guineapigs were mated overnight and the following day the vagina, uterus and oviduct were excised and washed with veronal acetate buffer pH 4.5, or 7 and/or 8 containing 0.9% NaCl . The washings contained be¬ tween 5 and 100 sperm in the uterus and tubes, more in the vagina. The sperm were smeared on a gelatin membrane and incubated overnight with buffer at pH 4.5 and 8.4. The areas of digestion were measured using a microscope, and a statistical analysis done for the sperm obtained from each organ. The results were compared with a similar experiment using epididymal sperm. The results indicate that the sperm proteases probably remain unchanged in the female genital tract up to the time of fertilization. THE VIRGINIA JOURNAL OF SCIENCE 189 HISTOLOGICAL EXAMINATION OF THE INFLAMMATORY RESPONSE TO CORNEAL IMPLANTS OF PHAGOCYTIC LEUKOCYTES. James W. Moore, III and Milton M. Sholley. Dept, of Anatomy, Virginia Commonwealth Univ. , Richmond, V A 23298. Inflammatory peri¬ toneal macrophages (M0) injected into the stroma of guinea pig corneas have been shown to induce vascularization of the normally avascular cornea. Neutrophils have been implicated as inducers of corneal vascularization in rats, although conflicting results have been obtained in other animals. Due to this controversy and the reported absence of infiltrating neutrophils in response to M0, we in¬ vestigated the potential of purified preparations of peptone elicited peritoneal M0 or neutrophils to induce vascularization in rabbit corneas. Our results show that autologous implants of elicited M0 induce vascular responses, whereas simi¬ lar implants of neutrophils do not induce vascularization.; Histological examina¬ tion at 4 to 24 hours revealed a significant infiltration of neutrophils from the limbal vasculature toward and reaching the M0 or neutrophil implant. Infil¬ tration was negligible at 48 hours and later times. These results show that au¬ tologous inflammatory M0 induce neovascular responses in the rabbit cornea and that there is early infiltration of the cornea by neutrophils. However, neutro¬ phils cannot be the inducing factor in vascularization, since vascular responses occured only to M0 implants. (Aided by NCI grant CA-26316) RECONSTITUTION OF THE IRON SUPEROXIDE DISMUTASE FROM BACTEROIDES THETAIOTAO- MICRON. Charles D. Pennington* and Eugene M. Gregory, Department of Biochem. and Nutr. , V.P.I., Blacksburg, VA 24061. Superoxide dismutase (SOD) from Bacteroides thetaiotaomicron has been purified 220 fold to apparent homogeneity as determined by HPLC and gel electrophoretic techniques. The specific acti¬ vity of the isolated SOD is 2200 U/mg. The enzyme has a molecular weight of 40,000 and is composed of two equal sized subunits joined by noncovalent interactions. The enzyme contains 2 gram atoms Fe and 1 gram atom Zn per mol. of dimer. Enzymatic activity is lost upon treatment with 5 M guanidini um Cl and 20 mM 8-hydroxyquinol ine. The denatured apoenzyme regains activity after dialysis against either 1 mM ferrous ammonium sulfate or .1 mM manganous chlor¬ ide in tris buffer. Ten yM zinc sulfate will inhibit Fe-reconsti tution by 78% and Mn-reconsti tution by 64%. The Fe-reconsti tuted and native enzyme have an apparent pH optimum of 6 whereas the apparent pH optimum of the Mn-reconsti tuted enzyme is 7.8. The native and Fe-reconsti tuted enzymes are inhibited 50% by .27 mM and .31 mM azide respectively. The Mn-reconsti tuted enzyme is inhibited 26% by 10 mM azide. (Supported in part by NIH Grant AI 1 5250-04 ) 1 SIALYLQLIGQSACCHARIDES OF HUMAN MILK: ISOLATION AND IDENTIFICATION BY ANTI- SIALYL OLIGOSACCHARIDE ANTIBODIES. Pedro A. Prieto* and David F. Smith, Dept, of Biochemistry, VA Polytechnic Inst, and State Univ., Blacksburg, VA 24061. Rabbit antiserum against the human milk oligosaccharide LSTb (Gal gl-3[NeuAca 2-6 ]G1 cNacgl -3Gal el -4G1 c) is used to isolate a new, structurally related oligo¬ saccharide. [^H] Si alylol i gosacchari des from milk of different Lewis blood group donors are separated by paper chromatography . The si alyl ol i gosacchari des are assayed for LSTb and related structures using anti -LSTb serum in a direct- binding radioimmunoassay. An oligosaccharide which moves slower than LSTb ! during paper chromatography is isolated. Based on its specific binding to anti- ; serum, its absence from Lea milk and its co-cromatography with lacto-N-fucopen- taose I (Fucal-2GalBl-3GlcNAc3l-3GalBl-4Glc) after neuraminidase digestion, the i proposed structure is: Fucal -2Gal Bl -3[NeuAca2-6]Gl cNacftl -3Gal el -4G1 c . 190 THE VIRGINIA JOURNAL OF SCIENCE STUDIES ON THE MECHANISM OF ACTION OF SOUTH AMERICAN 0-CARBOLINE HALLUCINOGENS. R, Mark Slusher* and Richard A. Glennon*, Department of Pharmaceutical Chemistry, MCV/VCU, Richmond, Virginia 23298. Harmala alkaloids, derivatives of 0-carbo- lines, appear to be the active constituents of a hallucinogenic decoction var¬ iously known as caapi or ayahuasca. The 3-carboline structure contains a rigid indolealkylamine (IAA) backbone. Various other IAAs are hallucinogenic in man and have been previously shown to interact with serotonin (5-HT) receptors in vitro; many of these IAAs are also active in tests of discriminative control of behavior in rats. We found that certain 0-carbolines are also active in this same paradigm. In order to gain further insight as to the nature of substituent effects, several novel derivatives were synthesized and evaluated using the drug discrimination paradigm. (Supported by USPHS grant DA-01642). DIETARY SUPPLEMENTATION WITH VITAMIN C DELAYS ONSET OF MUSCLE FATIGUE. J. Richardson, S_. Smith, and R. Allen*, Dept, of Biol. , Old Dominion University, Norfolk, Va. 23508. Prolonged vitamin C dietary supplementation increased con¬ traction time in isolated striated muscle but had no affect on the strength of the contraction. A total of thirty measurements were collected, fifteen from control and fifteen from experimental animals. The experimental animals were force fed 30 mg of vitamin C per day. Following the dosing period, the ani¬ mals were anesthetized and the gastrocnemius muscle was exposed. The leg and foot were immobilized by clamping and the Achilles tendon was attached to a "B" strength myograph which inturn registered muscle contractions on a physio- graph. The muscle was stimulated with an electrical probe using eight volts of 20 cycles per second until the muscle fatigued. THE EFFECT OF GLUCOSE DOSING ON MUSCLE FATIGUE. J. Richardson, S. Smith, and R. Allen*, Dept, of Biol., Old Dominion University, Norfolk, Va. 23508. After dosing with glucose, time to fatigue was measured in the gastrocnemius muscle in eleven experimental animals. Results indicate that contraction time was 23% longer for the dosed animals than for control animals. Ingestion of glucose thirty minutes prior to exertion increases stamina by prolonging muscle con¬ traction. Stamina was determined by anesthetizing the animals and exposing the gastrocnemius muscle. The Achilles tendon was attached to a spring loaded myograph of controlled tension. Muscle contraction was induced by an electri¬ cal probe using eight volts at 20 cycles per second. Time to fatigue was measured twice on the same muscle, but only the second contraction was used for data analysis. The muscle was allowed a 40 second recovery period be¬ tween stimulations. REGULATION OF MACROPHAGE (M0) la ANTIGEN EXPRESSION BY T CELL PRODUCTS. Lawrence B. Schook, Dept, of Microbiology and Immunology, Medical College of Virginia, VCU, Richmond, 'VA 23298. M0 were obtained from mouse bone marrow grown in the presence of colony stimulating factor (CSF). These M0 were phenotypically characterized for la antigen expression, ectoenzyme activities and effector functions during their in_ vitro differentiation. The kinetics of la antigen expression and tumoricidal activity of these M0 were disparate. Ia antigen ex¬ pression was CSF dependent and began at 3d of culture, was maximal after 5-7d, and then declined. M0 tumoricidal activity appeared after 7d, when the spon¬ taneous expression of Ia had declined. Treatment of M0 at 7d with lymphokines (LK) was able to maintain Ia antigen expression with no increase in tumor cell killing. Treatment with LK only increased tumor cell killing of BMDM obtained early during differentiation. Moreover, the more differentiated BMDM were less responsive to LK modulation of phenotypic characteristics indicating more stable phenotypes. (Supported by a grant from the American Cancer Society (IN-105G). THE VIRGINIA JOURNAL OF SCIENCE 191 SYNTHESIS, ANALYSIS AND BEHAVIORAL EFFECTS OF 2 , 3-METHYLENED IOXY AMPHETAMINE . W. H. Soine, R. Young* and R. A. Glennon, Dept, of Pharm. Chem. , MCV/VCU, Rich¬ mond, VA; R. E. Shark* and D. T. Agee*, Drug Examination Section, Bureau of Fo¬ rensic Science, Consolidated Lab. of Virginia, Richmond, VA 23219. Forensic and pharmacological differentiation of the methylenedioxyamphetamines (MDA) re¬ quired the synthesis of the previously unreported 2 , 3-methylenedioxyamphetamine which was converted to the nitropropene intermediate and then reduced with LiAlH^. For forensic purposes the isomers could be differentiated using the sulfuric acid color test, gas chromatography, mass spectrometry and ^C nuclear magnetic resonance. When I_ was tested in rats trained to discriminate (+)-3,4- MDA from saline in a two-lever drug discrimination task, generalization was found to occur, but I_ was 1/5 as active. Stimulous generalization did not occur with _I using rats trained to (+)-2 ,5-dimethoxy-4-methylphenylisopropylamine (DOM) or (+) -amphetamine. (Supported in part by NIDA grant DA-01642). INFLUENCE OF SEX, STRAIN AND SPECIES ON ALTERED TRACE METAL METABOLISM IN THE STZ-DIABETIC RODENT. Susan A. Spittle* and Mark L. Fail la, Dept, of Biochem. and Nutr., VA Polytechnic Institute and State Univ., Blacksburg, VA 24061. Previous studies have demonstrated marked alterations in trace metal metabolism in male SD rats following chemical induction of diabetes. To determine whether such changes represented a general response to the insulin-deficient condition the levels of Zn, Cu and Mn in liver (L), kidney (K), and intestine of normal and streptozotocin (STZ)-diabetic male rats of the Sprague-Dawley, Wistar and Long^Evans strains, female Sprague-Dawley rats and male mice were measured. Significantly increased concentrations of Zn, Cu and Mn in L and Zn and Cu in K were found in STZ-diabetic rats, regardless of sex and strain. In contrast, Zn and Cu contents in L and K of control and diabetic mice were similar, but Mn levels were significantly elevated in both organs of the diabetic mouse. Higher amounts of Zn and Cu were bound to metal lothionein in L and K of diabe- :i tic rats. (Supported by Pratt Nutrition Program and USDA). CELL MEDIATED ANTIGEN TRANSPORT TO LYMPH NODE GERMINAL CENTERS. Szakal, A.K. , : Schnizlein* C., and Tew, J.G. Dept, of Anat . and Microbiol., Med. Coll, of j Virginia/VCU, Richmond, Virginia. 23298. Long term antigen (Ag) retention in lymph node and spleen germinal centers (GC) by follicular dendritic cells (FDC) is well known. Evidence for the role of this phenomenon in the maintainance of long term immunity is mounting. The mechanism by which Ag arrives in lymph node IGCs is unknown. Hypotheses from passive drainage to active transport by lympho¬ cytes or macrophages have been proposed. The method of entry of Ag into GCs was systematically studied by light and electron microscopy in C3H mouse popliteal lymph nodes 1,5,15,30 minutes and 1,5, and 24 hours after foot pad injection of the Ag, HRP . Light microscopy showed an apparent migration of the peroxidase positive Ag on the afferent side of the lymph node from the subcapsular sinus (SS) at specific areas to regions of GCs. This occurred only in passively im- j munized controls indicating antibody dependence. Antigen migration into the cortex was already in progress at 1 min and formed continuous trails between j the SS and GCs between 1 min and 5 hrs. By 24 hrs the trails disappeared and HRP was localized in GCs. Electron microscopy showed that HRP was transported | to GCs on the surface of a group of cells of possible monocyte origin. Mor- I phology suggested that these cells may be precursors to FDCs. 192 THE VIRGINIA JOURNAL OF SCIENCE SYNTHESIS OF MESOIONIC THIADIAZOLOPYRIMID INE NUCLEOSIDES AS POTENTIAL ANTINEO¬ PLASTIC AGENTS. S. M, Tejani and R. A. Glennon*, Department of Pharmaceutical Chemistry, MCV/VCU, Richmond, VA 2329.8. 2 -Amino- 1 , 3, 4-thiadiazole (2-ATD) possesses activity as an antineoplastic agent. Although its exact mechanism of action is not yet fully understood, evidence suggests that it may undergo an exchange reaction with NAD to form a 2-ATD mononucleotide. A crude nucleotide metabolite (of unknown structure) of 2-ATD was isolated and demonstrated to be an inhibitor of IMP dehydrogenase obtained from leukemia L1210 cells. We syn¬ thesized 2-(D-ribof uranosy lamino) 1 , 3, 4-thiadiazole ; however it was found to be unstable. Mesoionic thiadiazolopyrimidine analogs of the above compound were prepared as potential prodrugs to modify the pharmacodynamic properties of the parent nucleoside and in an attempt to enhance stability. These potential pro¬ drugs might be predicted to slowly liberate the parent nucleoside in vivo. These compounds are currently being assayed for toxicity to L1210 and HEp-2 cells. (Aided by American Cancer Society Institutional Grant IN-105H). QUANTITATIVE IMMUNOHISTOCHEMISTRY OF PROLACTIN BINDING SITES IN VENTRAL PROS¬ TATES OF RATS TREATED WITH INDOMETHACIN . R.S. Vick*, J.R. Dave*, R.J. Witorsch*, Dept, of Physiology, Med. Col. of VA., Richmond, VA 23298. This study determined whether indomethacin (Indo) treatment had any effect on the immunohistology of prolactin (PRL) binding of rat ventral prostate (VP). Rats were injected with one of two doses of Indo for 0, 48, or 72h. VP sections were immunostained for PRL binding sites by the immunoperoxidase method. Immuno- staining was quantitated by two methods: the proportion of cells immunostained as measured by an image analyzer and the intensity of immunostaining per cell as measured by a microscope densitometer. Linear regression between the two para¬ meters showed a positive correlation at r = 0.75. The product of the two para¬ meters was used to estimate total PRL binding in VP. We found a significant in¬ crease in PRL binding with the low dose of Indo at 48h and significant decreases in PRL binding with the low dose at 72h and high dose at 48h. These immunohisto- chemical observations did not agree with our radioreceptor data which showed that Indo produced a time and dose dependent decrease in PRL binding. These con¬ trasting observations will be discussed. (NIH grant CA 23653). Characterization of Active and Inactive Forms of Rat Hepatic HMG-CoA Reductase. Terence R. Whitehead*, Z. Reno Vlahcevic, and Phillip B. Hylemon, Depts. of Microbiology and Immunology, and Medicine, Med. Coll. Va. , Va. Common. Univ. , Richmond, VA 23298. HMG-CoA reductase, the rate-limiting enzyme in cholesterol biosynthesis, was purified 4,500-fold with a specific activity of 8,930 nmoles/ min/mg, using agarose-HMG-CoA affinity chromatography. SDS-PAGE of the purified enzyme demonstrated one protein band. Additional protein isolated from the affinity column with 0,5 M KC1 demonstrated no HMG-CoA reductase activity, yet co-migrated with the active enzyme on SDS-PAGE. This protein was determined to be an inactive form of HMG-CoA reductase by tryptic peptide mapping, reaction with anti-HMG-CoA reductase antibody, and molecular sieving HPLC. Microsomal HMG-CoA reductase activity was found to be sensitive to the sulfhydryl inhib¬ itors PHMB and HgCl2, but not NEM. Both NADPH (4 mM) and HMG-CoA (176 jiM) were able to protect the enzyme against inhibition, indicating sulfhydryl groups are present in the active site of HMG-CoA reductase. THE VIRGINIA JOURNAL OF SCIENCE 193 MICROCOMPUTERS FOR PERSONAL INFORMATION STORAGE AND RETRIEVAL, Abund Ottokar Wist, Dept, of Physiology and Biophys. , MCV/VCU, Richmond, VA 23298. The rapid increase in knowledge and accompanying increase in the number of scientific papers, books, and other sources of information makes it increasingly difficult on the part of individual investigators to keep abreast of the litera¬ ture relating to their fields. Computer assisted literature searches are too comprehensive and often do not provide the desired information. Investigators on the frontiers of research seem to get the least benefit from such searches as a short review of literature search techniques shows. People with the least experience learn to use the library facilities most often. There¬ fore research workers with high experience set up their own literature search system which even though it is mostly manual fits better to their needs. Recently new technology such as mini- and microcomputers have been used to¬ gether with new search techniques to adapt the capabilities of a modern library more specifically to the needs of academic and medical research workers. Several such systems around the country will be described, together with a system we are developing here at the Medical College of Vi rginia/VCU. Microbiology Section BOVINE PARVOVIRUS ASSOCIATION WITH THE NUCLEAR MATRIX OF INFECTED CELLS. Laura L. Briggs* , Robert C. Bates, and Ernest R. Stout. Dept, of Biol., Virginia Polytechnic Institute and State Univ . , Blacksburg, Va. 24061. The nuclear matrix is a proteinaceous structure to which supercoiled loops of DNA are attached. The object of this research was to determine the nature of the association of Bovine Parvovirus (BPV) DNA and viral specific proteins with the nuclear matrix of infected cells. Matrices were prepared from virus- infected cells using two different procedures both utilizing non-ionic deter¬ gents and high salt conditions. Analysis of the DNA isolated from the matrices prepared from SV40- transformed bovine fetal lung cells infected with BPV show viral DNA to be associated with the nuclear matrix. Treatment of these pre¬ parations with increasing DNase concentrations ranging from 0 to 100 yg/ml does not decrease the amount of single-stranded BPV DNA associated with the matrix. Yet the amount of double-stranded BPV DNA does decrease with increasing DNase concentrations. SDS-poly acrylamide gel electrophoresis of matrix proteins demonstrates the presence of one viral capsid protein and one low-molecular weight, viral-coded nonstructural protein, NP1. The association of viral DNA and proteins with the nuclear matrix implicates a role for the matrix in the viral replication process. THE POLAR AND LATERAL FLAGELLA OF AZOSPIRILLUM BRASILENSE ARE ANTIGENICALLY DISTINCT. Patrick G. Hall & Noel R. Krieq, Dept, of Biol., Va. Polytech. Inst. & State Univ., Blacksburg, VA. 24061. An indirect immunoperoxidase stain was used to demonstrate by electron microscopy that an antigenic differ¬ ence exists between the polar flagellum and the lateral flagella of Azospir¬ illum brasi Tense ATCC 29145. No evidence could be obtained that the polar flagellum was ensheathed as it is in Vibrio spp. The diameter and wavelength of the two types of flagella are reported. 194 THE VIRGINIA JOURNAL OF SCIENCE DIMINISHED INTERLEUKIN 3 (IL-3) ACTIVITY IN TUMOR-BEARING HOSTS (TBH) . A.F . Lent ine, C.J. Burger, C.J. Denbow*, and K.D. Elgert . Biology Dept., Microbiology Section, Virginia Polytech. Inst. & State Univ., Blacksburg, VA 24061. IL-3 is produced during the generation of specific cytotoxic T lymphocytes (CTL) . A kinetic study of IL-3 production during tumor growth revealed a de¬ creased ability of TBH mouse splenocytes to produce IL-3. Removal of suppressor T cells did not restore IL-3 production in TBH splenocytes. Absorption studies suggested that fresh cells have more accessible IL-3 receptors than Con A- induced blast cells. Blastogenic assessment showed purified IL-3 was mitogenic for normal but not TBH splenocytes. IL-3 augmented the in vitro response of normal splenocytes to Con A and PHA but suppressed the TBH splenocyte response. Purified IL-3 inoculated into normal and TBH resulted in a suppression of TBH splenocyte blastogenesis . The further decrease in TBH splenocyte activity with exogenous IL-3 may be caused by tumor-induced feedback mechanisms that suppress cellular differentiation required for specific CTL generation. (Supported by NIH Grant CA25943 and a Whitehall Foundation Grant.) DNA/DNA HOMOLOGY STUDIES ON THE GENUS GLUCONOBACTER. Bruce Micales, J.L. Johnson and G.W. Claus, Virginia Tech, Blacksburg, Va. 24061. Since the present taxono¬ my of the genus Gluconobacter is based largely on phenotypic data, we investiga¬ ted them in terms of nucleotide sequence similarity. Using a SI nuclease pro¬ cedure, we determined DNA homologies from 39 strains of Gluconobacter , together with 30 strains of Acetobacter , Frateuria , and Pseudomonas . Three DNA homology groups were obtained with organisms generally fitting the description of the genus Gluconobacter. The average intergroup relatedness among the three homology groups was 20%. Type strains of the five subspecies of G_. oxydans all belonged to homology group I, with intra- and intersubspecies homologies ranging from 62 -100%. Homology group II was defined by reference strain IFO 3264 and showed an intragroup homology of 44-87%. Homology group III was defined by reference strain IFO 3276 and showed an intragroup homology of 68-99%. Our data suggests that the genus Gluconobacter is composed of at least three species and the current recognized subspecies are not justified. ASCITES FORMATION IN CHICKENS TRIGGERED BY RES OVERLOAD. Robert L. Taylor, Jr.*, Dept, of Pathology, Medical Col. of Va., Va. Commonwealth Univ., Richmond, Va. 23298, Imre Olah* and Bruce Glick*, Dept, of Poultry Science, Miss. State Univ., Miss. State, Miss 39762. Rapid ascites formation was observed in the hepatoperi- toneal sac 4 to 5 hours after IV injection of large amounts of carbon. The amount of ascitic fluid varied from 0.5 to 12 ml/chicken. The protein content of ascitic fluid was identical to that of serum. Total antibody titers to sheep red blood cells in ascitic fluid and serum were also similar. Specific IgM antibody in ascitic fluid was significantly lower than that in serum while the former showed more IgG antibody. The cellularity of ascitic fluid was poor consisting of mononuclear cells which were mainly macrophages with or without phagosomes. A few small lymphocytes and large numbers of thrombocytes were also present. In those animals with large amounts of ascitic fluid, caecal pouch contents were solid and relatively dry. These data suggest that a mechanism may exist which regulates water balance between the liver and the caecal pouch. THE VIRGINIA JOURNAL OF SCIENCE 195 MEMBRANE PROTEINS OF A METHANE-OXIDIZING BACTERIUM. Terry L. Weaver, Dept, of Biol., Liberty Baptist College, Lynchburg, V a. 24506. The methane oxidizing bacteria possess internal membrane vesicles regularly stacked in particular arrangements in the cytoplasm. The bacterium used in this study. Methyl osi nus trichospori urn, was grown in liquid under a methane/air (1/1) mixture, and after concentration the cells were disrupted using a Braun homogenizer. Internal membranes were isolated via differential centrifugation. Membrane protein samples were hydrolyzed in HC1 , and the polar amino acids were separated from the apolar by TLC. It was found that the amino acid composition of the membrane proteins was relatively enriched in apolar amino acids at over 71%. Electro¬ phoresis revealed five major membrane proteins with molecular weights of 47,000, 80,000, 125,000, 160,000, and 180,000. The membrane proteins were also analyzed spectrophotometrical ly for cytochromes via oxidized vs. reduced spectra and for the presence of enzymes using standard assay procedures. Cytochromes C and A, a cytochrome C reductase, and ATPase activities were all associated with the membranes. Thus the internal membranes in these bacteria appear to function primarily in electron transport and oxidative phosphorylation. THE EFFECT OF FRUCTOSE ON PROTEIN SYNTHESIS AND HETEROCYST FORMATION IN SEVERAL SPECIES OF CYANOBACTERIA. J. Ernest Wilson, Judith W. Scott, and Lisa C. Reekes , Dept, of Life Sciences, Va. State Univ., Petersburg, Va 23803. 5.0 mM fructose increases growth and protein synthesis in several species of cyanobacteria with heterocysts. Species which do not form heterocysts do not respond to fructose. When species that form heterocysts are grown in the presence of nitrate and am¬ monia which inhibit heterocyst formation, fructose does not increase growth or protein synthesis. If heterocysts are initially present in filaments treated with fructose and nitrate, a large initial increase in growth is found which disappears with time. Fructose increases heterocyst formation. It is suggested that these results would be obtained if fructose was the sugar which is normally transferred from vegetative cells to heterocysts. SENSITIVITY OF SEVERAL CELL LINES FOR INTERFERONS WHEN CHALLENGED WITH VSV AND EMC VIRUS. S. Yousefi*, C. W. Gouldin* and M. R. Escobar. Medical College of Virginia, Richmond, VA 23298. The clinical significance of quantitating IFNs in various human disorders has been confirmed. Among several criteria for choosing an IFN assay are simplicity, rapidity, cost-effectiveness, reproducibility, and sensitivity as well as its potential for automation. Diploid fibroblasts, de¬ spite their sensitivity, have inherent limitations. This problem led us to eval¬ uate monolayers of WISH, HEp-2, Vero, A549 and WI-26VA4 cell lines challenged with vesicular stomatitis virus (VSV) and encephalomyocarditis virus (EMCV) us¬ ing the conventional dye uptake assay for HuIFN-a and HuIFN-y. A549 and WI- 26VA4 did not meet these criteria. Vero cells challenged with EMCV were found to be suitable for HuIFN-a alone yielding high reproducibility, provided that sensitivity is not crucial. HEp-2 cells, readily available in the diagnostic lab. , challenged with VSV are very acceptable for both IFNs, particularly IFN-y. WISH cells with EMCV, instead of VSV, were found to be the most sensitive for both IFNs; however, these cells are not as readily available. A modification of the assay described above using WISH cells challenged with EMCV is more rapid without significant loss of sensitivity. It also lends itself to automation when screening large numbers of samples for HuIFN-a and HuIFN-y. 196 THE VIRGINIA JOURNAL OF SCIENCE Psychology Section PSYCHODIAGNOSTICS OF VOICE ANALYSIS: EXPERIMENTAL VALIDITY OF THE VOICE "STRESS-DISSECTION TEST." John C. Bartone, Department of Research, American Health Research Institute, Ltd . , ■ Annandale , Va. 22003. Voice analysis, per se, or attempts to "analyze" the voice by any form of stress evaluation, is a new field, inviting to all professions, and highly sensitive to differential errors unknown to all. Voice analysis is particularly applied in "lie detection" wherein proneness to errors is manifold. One ingenious experiment, the "Stress-Dissection Test," subject to universal application and adoption must be declared scientifically unsound, psychologically invalid and logically inapplicable. The parameters of the Stress-Dissection Test do not exist, have not been published and may contribute further confusion if applied consistently in the Lie Detection commercial arena. The Stress-Dissection Test is particularly dangerous in that clients, prisoners or patients may be given status that is against societal requirements. The high stress conditions under which this test operates is hazardous to clients, experimenters and outcome. THE EFFECT OF BILATERAL LESIONS OF THE NUCLEUS BASAL IS OF MEYNERT ON SPATIAL BEHAVIOR. Paul Chapman* Dept, of Psychology, Washington & Lee Univ. , Lexington, VA. 24450. (Sponsor: L. E. Jarrard) The effects of bilateral ibotenic acid lesions of the nucleus basalis of Meynert were tested in rats. This area has been shown to have important connections to neocortex as well as several limbic system structures. Two experiments were performed to examine the effects of damaging the nucleus on spatial memory. Since this nucleus has been implicated in Alzheimer's disease in humans, tasks were designed to test the animals working ("recent") memory, both on postoperative acquisition and retention of preoperative learning. The first of these involved memory in a radial maze on cued and non-cued tasks, and the second was a single alternation task. No reliable differences were found on either type of learning. The results of these experiments suggest that the inputs from the nucleus basalis, particularly to the limbic system, have little or no importance in spatial learning or retention. RESPONSE BIAS IN A EIXED INTERVAL SCHEDULE. R. Eeguson, P. Martin, and M_. Griffin , Dept, of Psychology, Va. State Univ., Petersburg, Va. 23803. Responding under Fixed Interval (El) schedules typically begins after only about 50 percent of the interval has elapsed. This has been attributed to a schedule bias: late responses are punished by delaying reinforcement while early responses are relatively effortless. The purpose of this research was to test this view by making early responses more "expensive". Twelve Holtz- man male rats were initially trained on an FI60 sec schedule to which was than added a loud buzzer for .5 sec immediately following each response. Statistical analyses indicated no significant differences between the res¬ ponse functions under baseline and buzzer conditions. Subsequent research has demonstrated that the buzzer is an aversive stimulus and that the absence of a buzzer effect in the El data can not be attributed to stim¬ ulation deafness. These data thus call into question the adequacy of the schedule bias account of El responding. This work was supported by NIH/MBS Grant No. RR 08090-11. THE VIRGINIA JOURNAL OF SCIENCE 197 BEHAVIORAL CONTRAST AND TEMPORAL DISCRIMINATION. R. Ferguson, P. Martin, and S^ Lambert , Dept, of Psychology, Va. State Univ., Petersburg, Va . 23803. Responding under fixed interval (FI) schedules of reinforcement has been characterized as representing a temporal discrimination process in which different temporal points in the interval serve as discriminative stimuli. The purpose of this experiment was to test this view by attempting to demon¬ strate behavioral contrast along a strictly temporal dimension. Twelve male Holtzman rats were trained on a discrete trial mixed FI30-EI60 sec schedule and then shifted to a discrete trial mixed EXT-FI60 schedule in which the 30 sec trials terminated with lever retraction but no reinforcement. The data from the second phase of the study showed clear evidence of contrast: responding during sec 20-30 of each trial decreased across the 15 test days while responding during sec 50-60 increased. These data thus strongly suggest that time serves as a discriminative stimulus in termporal schedules of reinforcement. This work was supported by NIH/MB5 Grant No. RR- 08090-11. THE EFFECTS OF UNILATERAL AND BILATERAL ENTORHINAL LESIONS ON SPATIAL MEMORY IN THE RAT. H. E. Gill* G. K. Mauser* and _E. D. Vaughan* Dept, of Psychology, Washington & Lee Univ., Lexington, VA. 24450. (Sponsor: L. E. Jarrard) The effects of unilateral and bilateral entorhinal cortex (EC) lesions on retention of a complex spatial task were studied. Rats were initially trained | in the Morris water task. This task consists of a circular pool filled with opaque water where the subjects have to find safety on a platform located j just below the water. After learning to locate the hidden platform, j unilateral, bilateral, and sham EC lesions were performed. Three days following the operations testing on the task was resumed. Unilateral lesioned rats were slightly impaired but the results were not statistically different from the operated controls. Bilateral lesioned rats initially showed a large deficit in performance, but performance gradually improved over trials. The results will be discussed as they relate to synaptogenesis and recovery of function. THE USE OF THE AWARENESS SESSION IN MOTIVATING SEAT-BELT USAGE: THE APPLICATION ! OF SOCIAL PSYCHOLOGICAL PRINCIPLES IN AN INDUSTRIAL SETTING. Heidi Ann Hahn* & E. Scott Geller, Dept, of Phych. , Va. Polytechnic Inst. & State Univ., Blacksburg, Va., 24060. Research has shown that seat-belt wearing among white- collar workers can be motivated through the use of incentives. Incentive strategies have not, however, been effective among blue-collar workers. Using the principles of social psychology, an "awareness session", that is, a group discussion technique designed to elicit involvement and commitment on the part of blue-collar workers, was developed. The current study evaluated a program which combined this "awareness session" technique with an incentive strategy similar to that used in previous research. The results indicated that this combination was very effective. in motivating safety belt use among blue-collar workers, effecting an eight-fold increase in the percentage of seat-belt wearers. Implications regarding the use of social psychological principles in research in behavioral community psychology are discussed. 198 THE VIRGINIA JOURNAL OF SCIENCE FINE MOTOR CONTROL AND THE EFFECTS OF BASAL GANGLIA LESIONS IN THE RAT. John V. Harrell, Dept, of Psych., Hampden-Sydney Col., Hampden-Sydney , Va. 23943 The basal ganglia are thought to perform a ramp generation function necessary for control of slow movement, making it possible to refine the movement as it is being performed. To examine such a role for the basal ganglia in the rat, one group received bilateral caudate nucleus lesions and another group received control lesions in parietal cortex. The effects of these lesions were ascer¬ tained using a force-band/duration task. The task required the rats to limit the force of forelimb paw presses on a force transducer to a range of 20-50 gm and to hold this limited force response for a minimum of 1.5 sec in order to produce water reinforcers. Responses in which the duration requirement was not met or in which the force emission strayed from the preset range reset all response criteria. Results of the study indicate that rats with caudate nucleus lesions had extreme difficulty in limiting their force emissions to the required range of force but retained their ability to make the required response duration. ESP IN THE CLASSROOM: PAST AND PRESENT. Ann W. Hill, Math Dept., Smithfield High Sch., Smithfield, VA 23U30. This project was designed to achieve three goals: (1.) to get marked evidence of ESP from my candidate; (2.) to determine the extent of my participant's ability to display evidence of ESP by the old and new methodologies; (3.) to determine whether or not 'time' had an effect on ESP results if the psychological conditions were equal. There are students in our classrooms who possess extrasensory perception. My conjecture is that these students are special persons who have some kind of permanent, stable personality characteristic and mental attitude. My ultimate goal is to uncover these abilities and channel them into areas useful to the individuals. SPATIAL LOCALIZATION IN THE MOUSE. L. E. Jarrard, Dept, of Psychology, Washington & Lee Univ. , Lexington, VA. 24450, and A. Badgett*, L. Draper*, & H. Jarrard*, Lexington High School, Lexington, VA. 24450. In order to see if mice can learn to locate an object that remains in a fixed location relative to diptal room cues, animals were trained in a Morris swimming task. The task consists of having the subjects swim in a circular pool filled with opaque water in order to find a "safe" platform. In the Place Condition the platform was in the same location from trial to trial just below the water surface thus eliminating visual and possible olfactory cues. In the Cue Condition the platform was visibly above the water surface but location varied over trials. Learning occurred rapidly in both groups of mice but significantly slower in the Place Condition. In order to see if the animals in the Place Condition were using spatial localization, a transfer test was carried out where the platform was moved to a new location. There was a resulting increase in swimming time thus demonstrating that the mice were not using local cues to find the platform. The data will be discussed in relation to recent theorizing concerning spatial memory in animals. THE PaTE REPORT, A PSYCHODYNAMIC AND THERAPEUTIC EVALUATION. Lois S. Kriegman, The Kriegman Clinic, Ltd., 106 N. Thompson St., Richmond, Va. 23221 & George Kriegman, M.D.* (deceased). This is a report of a procedure which had its origins in the authors' collaborative search for new techniques which can accel erate the therapeutic process and shorten the period of therapy necessary for the patient, whereby the emotional impairment and the economic loss to the indi vidual, his family, and his community would be diminished. It was felt that if a quick penetration in depth into the basic core of the personality structure could be achieved, psychotherapeutic efforts could be more pointedly directed and the initial exploratory period in therapy diminished. The PaTE Report is a systematic psychological reporting procedure based on a testing technique and a particular theoretical orientation. It is an attempt to achieve the goal of a rapid psychodynamic and therapeutic evaluation of emotional problems. This report, derived from the scores of the Wechsler Intelligence Scales and Kahn Test of Symbol Arrangement, has been in constant use by the authors since 1959. THE VIRGINIA JOURNAL OF SCIENCE 199 INDIVIDUAL AND COMBINED EFFECTS OF PHENCYCLIDINE, ETHANOL, AND NICOTINE ON LETHALITY IN RATS. P. M. Martin and W ,T . Hodge , Dept, of Psychology, Va . State Univ., Petersburg, Va . 23803. The purpose of this experiment was to examine the lethality of PCP alone and in combination with two commonly used psychoactive drugs, ethanol and nicotine. Rats were given either 100, 125, or 150 mg/kg of PCP and the number of animals that died within 24 hours was recorded. The median lethal dose LD50) for PCP was 127 mg/kg. The LD50 for PCP was then recalculated for rats predosed with nonlethal doses of ethanol or nicotine. When animals were predosed with ethanol (1000 mg/ kg), the LD50 of PCP was 100 mg/kg, while the LD50 of PCP was 106 mg/kg for animals predosed with nicotine (60 g/kg). These were both significantly lower than the LD50 for PCP alone. These results show that both ethanol and nicotine enhance the lethality of PCP, thus suggesting that some of the toxic or lethal effects of PCP may be due to its interaction with other psychoactive drugs. This research was supported by NIH/MBRS Grant No. RR- 08090- 11. A BEHAVIORAL PROFILE OF PHENCYCLIDINE IN THE RAT. P. Martin*, D. Turner* W. Hodge and R. Ferguson*. Dept, of Psych, Va. St. Univ., Petersburg, Va. 23803. Since phencyclidine hydrochloride (PCP) acts as a dopamine agonist as do amphet¬ amine and apomorphine, the purpose of the present experiment was to develop a behaviocal_prof ile for PCP which could be compared to those of amphetamine and apomorphine. Rats were given isotonic saline or 4, 8 or 16 mg/kg of PCP i.p. before being placed in an unfamiliar test cage. The frequency and duration of 10 behaviors were recorded using a double blind procedure during a 1 hr test session. PCP caused dose and time dependent decreases in sniffing and rearing and increases in swaying, locomotion and rotation. Although amphetamine and apomorphine increase locomotion and cause rotation as did PCP, these stimulants also increase, rather than decrease, rearing and sniffing behaviors in animals who have been adapted to the test chamber. Thus, the differences between these drugs may be due to either (a) different mechanisms of action or (b) an inter¬ action between the drug and familiarity with the test environment. Rats who have been adapted to the test chamber are now being tested with the same doses of PCP to determine if such an interaction exists. This research was supported by NIH/MBRS Grant Number RR-08090-11. OPERATIONAL DEFINITIONS: HELP OR HINDRANCE IN A MATURING SCIENCE? John L. Miles, Jr., U.S. Army Research Inst, for the Behavioral & Social Sciences, Alexandria, VA 22333. The concept of "operational definition" is taught to students as a means of aiding them in thinking clearly and expressing themselves in a way that can be readily understood by others who apply the scientific method. While thus a potential boon to certain scientists — particularly those who may with to replicate a particular experimental situation — the existence of operational definitions can also have a divisive and retarding effect on the development of new technologies. This paper reviews the practical effects of proliferation of operational definitions within an area of psychology which describes human be¬ havior in the form of "task analysis." 200 THE VIRGINIA JOURNAL OF SCIENCE USING BIBLIOGRAPHIC TOOLS FOR INDIVIDUALIZING PSYCHOLOGY INSTRUCTION. James P^. B. O'Brien, Div. of Social Sciences, Tidewater Cmnty. Col., Virginia Beach, VA 23456. Our pursuit of scientific research normally begins with a careful study of the literature. While we usually begin instruction in scientific method in an undergraduate's first course, developing the skillful use of the scientific literature is less systematic and provided piecemeal at the graduate level when presented at all. If our journals, handbooks, abstracts, and other reference works are vital in defining our problems, guiding our research, and organizing the knowledge of our discipline; it follows that their early pur¬ poseful introduction to students would be beneficial. The skills developed in using such basic tools will not only enhance their understanding of a particu¬ lar science, but also will provide them with access to information useful in their daily lives. The argument is made that teaching these skills should not be delayed until the senior project or graduate school; nor should they be left to chance if we seek a wider, more cogent public awareness of science. Several examples in class handouts for a 3-quarter General Psychology course are dis¬ tributed and discussed. Numerous learning principles including successive ap¬ proximation, feedback, active student involvement, and the fostering of student initiative and responsibility are applied. DEVELOPMENT OF A SUPPORT GROUP FOR FAMILIES OF STROKE PATIENTS . Ann R. Penberthy, Sandra Perosa,* McGuire VAMC, Richmond, VA 23249 While patients recovering from stroke have noted difficulties in adjusting to the significant life changes associated with sudden and debilitating decrements in physical abilities, less attention has been paid to the parallel difficulties experienced by family members adjusting to concomitant changes in the family structure. Weekly meetings are held for emotional support of the families while the patients receive a concurrent speech therapy hour for practice of communica¬ tion skills. As the support group developed, distinct patterns to the adjustment of family members were apparent. Discussion of these expected patterns with the new families joiningthe group has assisted in reduction of problems and a faster adjustment from the self-reports. Coping strategies utilized by the participants are discussed, and families have proved to be quite creative in developing a number of helpful activities to address the problems encountered. Research strategies to evaluate the impact of the support group activities on patient out¬ come variables and adjustment patterns of patients and family members were dis¬ cussed. Implications for the development of similar support groups were con¬ sidered . THE CHALLENGE GROUP: COPING WITH CHRONIC ILLNESS. Ann R. Penberthy, Linda Perosa, and Marlene Dmy tryshyn , * McGuire VAMC, Richmond, VA 23249 The development of an inpatient open-ended support group for elderly chronically ill patients was described together with the research plans underway to obtain empirical data on the reports of improvement in adjustment, attitudes, and other psycho-social variables attributed to the impact of group therapy. The problems encountered in conducting groups on the medical wards include: staggered timing of admission and discharge, scheduling conflicts with OT, PT, RT, medical consults and family visits, memory losses for recent stroke patients. To accommodate these multiple disruptions, the groups leaders allocate several hours each week to scheduling arrangements, posting of names, and personal reminders to recently im¬ paired group members. As a result, the group has been successfully maintained now for several months. Patients report that the discussions have been helpful in adjusting to the severe changes precipitated by sudden changes in health status, and associated reactions to decrements in chronic illnesses. Although the group size and composition changes weekly with the admissions and discharges, current members apparently develop a social support network that continues on the ward and facilitates adjustment of new patients. THE VIRGINIA JOURNAL OF SCIENCE 201 INCENTIVES AND SAFETY BELT USE: THE PROBLEM OF RESPONSE MAINTENANCE. James R. Rudd* & E. Scott Geller , Dept, of Psych., Virginia Polytechnic Inst., Blacksburg, Va. 24060. In the U.S. more than 34,000 deaths occur each year to occupants of passenger cars, light trucks, and vans. Safety belt usage could reduce the chance of injury by 50% or more. Unfortunately, the rate of safety belt use in this counj try is only 10%. Because of this low usage rate, the National Highway Traffic Safety Administration (NHTSA) formed an interdisciplinary committee to evaluate strategies for motivating seat belt use, which has included incentive programming Incentive programs, which are based on reinforcement theory, have proven excep¬ tionally successful in increasing safety belt use --increases of 100% or more are not uncommon. However, when the incentive programs are withdrawn, belt usage often returns to baseline levels. This paper is a theoretical discussion of pos¬ sible solutions to this problem of response maintenance. Ongoing programming CSTEP theory) and commitment vs. incentive programs are discussed along with re¬ lated factors such as observer bias, the Hawthorne Effect, and treatment packaged THE DRUNK-DRIVING PROBLEM « TOWARD A COMMUNITY- BASED PROGRAM. Ann Ei Talton V Mark AltomarH and E. Scott Geller, Dept, of Psychology, Va, Polytach. Inst,, Blacksburg, Va, , 24061 , Efforts to reduce alcohol- Impaired driving have traditionally focused on education, legislation, and rehabilitation. To date, such strategies have been relatively unsuccessful. In an effort to depart from traditional interventions , an assessment plan was designed to gauge the drunk-driving problem in a university community. As a first step, several questionnaires were con¬ structed and administered to 355 students at Va. Tech. These were designed to tap students* opinions and attitudes toward various aspects of alcohol use in college and particularly to Issues related to drunk-driving. Results will be ^c^sed along with Implications for comprehensive communitywide interventions to reduce drunk-driving. Statistics Section f; TWO-TAIL TESTS vs. TWO- ALTERNATIVE TESTS. Robert L. . Andrews, Dept, of I' Business Administration, Virginia Commonwealth University, Richmond, VA 23284 i We are creatures of habit. We often are not open to examining procedures that differ from those that we were taught. When we do examine something different we carry a lot of baggage and prejudice into the examination. This paper compares the customary one-tail or two-tail hypothesis tests with a two-alternative procedure. The comparison emphasizes the perspective of the student who has not been biased by previous education. The author finds the two-alternative test to be a superior procedure. The final question is, "Which procedure should we teach?" NONPARA METRIC ONE -SAMPLE BAYESIAN INFERENCE. David L. Banks, Dept, of Statistics, Va. Polytechnic Inst., Blacksburg, Va., 24o6l . Suppose one wishes to perform a one-sample hypothesis test about a general parameter when the underlying distribution is known to have finite support on a given interval. In this situation it is possible to develop a nonparametric test that adheres to the principles of sufficiency, conditionality and likelihood, and which permits the statistician to employ prior information in a flexible manner. The development depends on the Imposition of a measure on a suitable subset of the space of measures; for best results, this prior measure should combine a roughness penalty with an appropriate weighting scheme. This leads to a persuasive Bayesian test, and there are immediate multivariate extensions. 202 THE VIRGINIA JOURNAL OF SCIENCE NORMAL TOLERANCE LIMITS: A STUDY OF ROBUSTNESS. George C. Canavos, School of Business, Virginia Commonwealth University, Richmond, VA 23284. Robustness pro¬ perties of the two-sided tolerance limits for normal distributions are examined based on a computer simulation in which the Student’s t and gamma distributions are used as generating models. Results indicate that the normal tolerance limits are sensitive to departures from normality when k-f actors are selected for cove¬ rages in excess of 0.9. Reasonable robustness is achieved for coverages of at most 0.9, as long as the underlying distribution is not extremely long-tailed nor highly skewed. A CONFIDENCE INTERVAL ON THE RESPONSE AT THE OPTIMAL LEVELS OF THE INDEPENDENT VARIABLES IN A RESPONSE SURFACE. Walter H. Carter, Jr.*, Vernon M. Chinchilli*, Eleanor D. Campbell*, Dept, of Biostat. , V a. Commonwealth Univ., and Galen L. Wampler*, Dept, of Medicine, V a. Commonwealth Univ., Richmond, V a. 23298. This paper presents a method for calculating a confidence interval about the response at the stationary point of a response surface. It is shown that the technique can be used also to calculate a confidence region about the location of the stationary point. The procedure is applied to the analysis of survival data via Cox's proportional hazards model of two preclinical cancer therapy experiments. The first experiment involves a single drug while the second evaluates a combination of two drugs. The results are discussed in terms of treatment activity and therapeutic synergism. ANALYSIS OF COVARIANCE IN THE MULTIVARIATE GROWTH CURVE MODEL. Vernon M. Chinchilli, Dept, of Biostatistics, Va. Commonwealth Univ. , Richmond, Va. 23298 . The multivariate growth curve model fits a polynomial growth curve for repeated measurements of a response variable. If the experimental units have been randomized to k treatment groups, then k distinct polynomial curves of the same degree can be inserted into the model. Usually it is of interest to com¬ pare statistically the k growth curves. In some experimental situations it may be necessary to adjust for covariables. The multivariate growth curve model can adjust for covariables by categorizing them into m strata and then model¬ ling mk polynomial curves. This results in too many parameters for estimation and too few error degrees of freedom for hypothesis testing. An extension of the multivariate growth curve model has been developed which permits the use of covariables in a manner analogous to the classical MANCOVA model. SIMULTANEOUS RANDOMIZATION TESTS. Robert V. Foutz*. Dept, of Stat. , Va. Polytechnic Inst., Blacksburg, VA 24061. Randomization analyses have been developed for testing main effects and interactions in standard experimental designs. However, exact multiple comparisons procedures for these randomiza¬ tion analyses have yet to be treated. This article proposes a general pro¬ cedure for constructing simultaneous randomization tests that have prescribed type I error rates. An application of the procedure does provide for multiple comparisons in the randomization analyses of designed experiments. This ap¬ plication is made to data collected in a biopharmaceutical experiment. THE VIRGINIA JOURNAL OF SCIENCE 203 THE EFFECT OF BIAS, VARIANCE ESTIMATION, SKEWNESS, AND KURTOSIS OF THE EMPIRICAL LOGIT ON WEIGHTED LEAST SQUARES ANALYSES. John J. Gart, Hugh M. Pettigrew, and Donald G. Thomas, Biometry Br. , National Cancer Institute, Bethesda, MD. 20205. The first four cumulants of empirical logit transformation of binomial variates are studied exactly and in asymptotic expansions. The covariance of the enpirical logit and its estimated variance is also considered. The ± 1/2 corrections are found to be not universally effective in reducing bias. Other corrections are suggested for simple logistic regression and the estimation of a common odds ratio in several 2x2 tables. The distribution of the enpirical logit is more skewed and has a larger kurtosis than the binomial. Confidence interval methods based on binomial variates, such as Cornfield's limits for the odds ratio, have better properties than empirical logit methods. Methods based directly on sufficient statistics or functions thereof, such as the maximum likelihood estimators corrected for asymptotic bias, are recommended. USE OF CONFIDENCE REGIONS FOR TESTING HYPOTHESES IN CLINICAL TRIALS. Janis L. Goodlow* , Dept, of Biostatistics, Med. Col. of Va., Va. Commonwealth Univ. , Richmond, Va 23298. At the present time combination chemotherapy may be less effective than it is cap¬ able of being. Current methodology depends on the investigator making an educated and fairly accurate estimate as to what the op¬ timal treatment level (s) are. Presented here is a new approach that does not rely as heavily on an investigator’s intuition. This approach is a combination of survival analysis, response sur¬ face analysis and design of experiments. The effects on the proce¬ dure of varying the conditions upon which the assumptions are based are investigated. This procedure has the potential of yielding in¬ formation concerning drug action and interaction that may aid in the development of improved clinical protocols. (Aided by Grant IN-105H from the American Cancer Society.) COLLINEARITY IN SIMULTANEOUS SYSTEMS. William P. Grubbs & Oral Capps, Jr., Dept, of Stat . , Va. Polytechnic Inst., Blacksburg, Va 24061. Many situations in quantitative economic analysis cannot be modeled under the principle of uni¬ directional flow of causality, as is common with many types of analyses in the physical sciences. Simultaneous systems of equations are commonly used to de¬ scribe economic phenomena. Simultaneous systems are classified according to the concept of identification, and many systems are classified as over-identified. A popular technique for dealing with over-identified systems of equations is Two Stage Least Squares (2SLS) . As least squares suffers from imprecise estimation when ill-conditioned data have been collected, so does 2SLS in a simultaneous system framework. Techniques have been proposed to circumvent ill-conditioning, using variants of the popular ridge regression and principal component regression procedures, as adapted to 2SLS. It is hoped that these estimators have smaller mean squared error than the 2SLS estimator. Simulation studies involving ridge regression in both stages of 2SLS are being conducted. LOG-LINEAR MODEL ANALYSIS OF THE ASSOCIATION BETWEEN DISEASE AND GENOTYPE. K. Hinkelmann, Department of Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061 & H.J. Khamis? Department of Statistics, Wright State University, Dayton, OH. Further insight into the analysis of the disease-genotype association problem, as discussed by Norwood and Hinkelmann (1978), is provided through the use of the log-linear model. It is shown how this approach can take the structure of the data into account for testing hypotheses about the type of association, specifying the form of recurrence risks, and estimation of such recurrence risks by maximum likelihood. The notion of conditional recurrence riqk is introduced and its usefulness is illustrated. 204 THE VIRGINIA JOURNAL OF SCIENCE VARIANCE REDUCTION TECHNIQUES USING RESPONSE SURFACE DESIGNS. James R. Hussey & Raymond H. Myers, Dept, of Statistics, Va. Polytechnic Inst., Blacksburg, Va. 24061. In computer simulation studies, the experimenter can control conditions in the system. One example is the ability to induce positive and negative correlations among the observations, rather than using independent observations. This can be done using the techniques of common variates and antithetic variates. These correlations alter the variance-covariance matrix of the design employed in the experiment. Although many possible correlation structures exist, certain patterns emerge as being more beneficial than others. These patterns can be viewed as the design being separated into blocks, where the correlations define the blocking. With these patterns, variance reduction is realized in many cases, especially in designs which admit orthogonal blocking. Variance is measured by prediction variance, integrated variance, and other criteria. Common response surface designs of first and second order are considered, with generally favor¬ able results. CELL MEANS ANOVA OF TWO-WAY LAYOUTS: A GEOMETRIC VIEW. Robert E. Johnson, Dept, of Mathematical Sciences, Va. Commonwealth Univ., Richmond, Va. 23284. Yates (1934) presented a method of analysis of two-way, fixed effects designs using as the one observation per cell the estimated cell mean and derived an approximate F test utilizing an adjusted mean squared error. This procedure, known as the unweighted means analysis, can be seen as a quite natural one when viewed geometrical ly . Via the geometric approach, conditions for independence of sums of squares and for exact F-di stri butions of variance ratios will be given in terms of some measures of nonorthogonality. DISPUTED PATERNITY. Stephen P. Mandel^ Department of Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061. The follow¬ ing situation arises alarmingly often: a lady claims that Mr. X is the bio¬ logical father of her child; Mr. X says that he is not. This leads to a dis¬ pute that sometimes can only be settled in the law courts. Counsel for Mr. X would first try to clear his client by attempting to demonstrate a genetic incompatibility between Mr. X and the child. For instance, if he has blood group 0 and the child has AB then he could not have sired that child and would (now) be acquitted on the basis of that evidence. The ramifications will be discussed. REDUCING TRANSFORMATION BIAS IN CURVE FITTING. Don M. Miller, School of Busi¬ ness, Va. Commonwealth Univ., Richmond, Va. 23284. A variety of methods exist for modeling a non-linear relationship between two variables. For what is probably the most common method in practice, linear regression analysis is ap¬ plied to the variables after they have first been linearized by a transforma¬ tion. The curve is then obtained by applying the inverse transformation to the fitted line. However, this procedure produces what can be a serious bias. This bias is identified for each of a simple set of transformations which provide satisfactory linearization for a large number of applications. Simple adjust¬ ments are presented which greatly reduce these biases. THE MULTIPLE DESIGN MULTIVARIATE LINEAR MODEL. Barry H. Schwab* , Dept, of Biostatistics, Med. Col. of Va., Richmond , Va. 23298. The multiple design multivariate (MDM) linear model can be applied to a multivariate regression system with polynomial models of different degree in the dependent variates. Zellner (1962) con¬ sidered a method of estimating the parameters of a multivariate regression system falling under the MDM model. The procedure applies Aitken's generalized leas t - squares to the whole system of equations to simultaneously estimate the regression coefficients. I will investigate whether the estimation technique can be im¬ proved by performing an iterative extension to Zellner* s least- squares algorithm. THE VIRGINIA JOURNAL OF SCIENCE 205 STOCHASTIC FLOW SHOP WITH BLOCKING. S. Suresh, Dept, of Industrial Engineering and Operations Research, Va. Polytechnic Inst. & State Univ. , Blacksburg, VA 24061. Pinedo has conjectured that in a stochastic flow shop with m machines, n-2 deterministic jobs with unit processing time, and two stochastic jobs with mean one, the optimal sequence to minimize the expected makespan is to schedule one of the stochastic jobs first and the other last. We prove that the optimal sequence is either the sequence of Pinedo’ s or the sequence in which the stochastic jobs are adjacent at one end of the sequence. CONFIDENCE INTERVALS FOR CEP ESTIMATORS WITH ELLIPTICAL NORMAL ERRORS. Audrey E. Taub* and Marlin A. Thomas* Mathematical Statistics Staff, Naval Surface Weapons Center, Dahlgren, VA 22448. A common parameter for describing the accuracy of a weapon is the circular probable error, commonly referred to as CEP. CEP is defined as the radius of a mean centered circle within which 50% of the future rounds from a particular weapon system will fall. If the fall of shot follows an elliptical normal distribution, a number of estimators exist for estimating CEP. This paper develops approximate confidence intervals for each of five commonly used CEP estimators. NOTES Mr- peter M. Mazzeo ’w|' s- National Arboretum Washington, 0c 20002 THE VIRGINIA JOURNAL OF SCIENCE EDITOR: Stewart Ware Dept, of Biology College of William and Mary Williamsburg, Va. 23185 BUSINESS MANAGER: N. J. Fashing Dept, of Biology College of William and Mary Williamsburg, Va. 23185 ©Copyright, 1983 by the Virginia Academy of Science, The Virginia Journal of Science (ISSN 0042-658X) is published four times a year (Spring, Summer, Fall, Winter) by the Virginia Academy of Science, Department of Biology, University of Richmond, Richmond, Va. 23 1 73. Second Class Postage paid at Richmond, Virginia and additional entry at Harrisonburg, Virginia 22801. The Virginia Academy of Science and the Editors of the Virginia Journal of Science assume no responsibility for statements or opinions advanced by contributors. Subscription rates for 1983: $27.50 per year, U.S.A.; $35.00 per year, other countries. All foreign remittances must be made at par U.S. dollars or their foreign equivalent. Back issues are available for $7.50 per issue postpaid. Changes of address, including both new and old zip codes, should be send promptly upon moving to the following address. POSTMASTER: Send address changes to Blanton M. Bruner, Executive Secretary- Treasurer, Virginia Academy of Science, Department of Biology, University of Richmond, Richmond, Va. 23173. All correspondence relating to remittances, advertising (rates will be sent upon request), subscriptions, missing issues, and other business affairs should be addressed to Business Manager. Instructions to Authors All manuscripts and correspondence about them should be addressed to the Editor. The Virginia Journal of Science welcomes for consideration original articles and short notes in the various disciplines of engineering and science. Cross-discriplinary papers dealing with advancements in science and technology and the impact of these on man and society are particularly welcome. Submission of an article implies that the article has not been published elsewhere while under consideration by the Journal. The original and two copies of each manuscript and of all figures therein are required. Authors should submit names of three potential reviewers. All manuscripts must be double-spaced through¬ out. The title, author’s name, affiliation and address should be placed on a covering page. An abstract (on a separate sheet) summarizing the text, particularly the results and conclusions, is required. After revision and final acceptance of an article, the author will be asked to furnish an error-free, camera-ready copy of the manuscript typewritten single-spaced with a black carbon ribbon on white bond paper. The text should follow the general format used by professional journals in the author’s disci¬ pline. Literature cited in the text should follow the name-and-year format: Fujishima and Honda (1972), or Spry (1969), or Guliday ( 1971). On the Literature Cited page at the end of the article each reference should include author(s), year, title of article, title of journal (using standard abbrevia¬ tions), volume number, and first and last page of the article. For a book, include author(s), year title, pages or number of pages, and publisher and city of publication. Examples: Fujishima, A., and Honda, K. 1972. Electrochemical Photolysis of Water at a Semiconductor Electrode. Nature 238: 37-38. Spry, A. 1969. Metamorphic Textures. Pergamon Press, New York. 350 pp. Guliday, John E. 1971 . Pleistocene History of the Appalachian Mammal Fauna. In Distributional History of the Southern Appalachians, Part Ill. Vertebrates (Perry C. Holt, ed.). pp. 223-262. VP1 & SU, Blacksburg, Va. Each figure and table should be mentioned specifically in the text, with all figure numbers and legends typed consecutively on separate sheet, and the figures identified by author’s name and figure number in pencil on the back. Table numbers and legends should be included as part of the table. Authors will be allowed 15 printed pages (including figures) free, but payment of $25 per page will be charged for the 16th and subsequent pages. VIRGINIA JOURNAL OF SCIENCE OFFICIAL PUBLICATION OF THE VIRGINIA ACADEMY OF SCIENCE Vol. 34 No. 4 WINTER 1983 TABLE OF CONTENTS ARTICLES Intergradation Among Ringneck Snakes, Diadophis punctatus (Linnaeus) in Virginia. Charles R. Blem <& Cheryl Roeding, Virginia Commonwealth Univ. 207 Phytoplankton Studies Within the Virginia Barrier Islands. III. Phytoplankton Composition in a Saline Pond on Smith Island. Harold G. Marshall, Old Dominion Univ. 215 Introduction of Anthribus nebulosus (Coleoptera: Anthribidae) in Virginia for Control of Scale Insects: A Review. Michael Kosztarab, V.P.I. & S.U., and Ferenc Kozar, Hungarian Research Institute of Plant Protection 223 Food Consumption, Mating Behavior, and Shelter Selection of Anthribus nebulosus (Coleoptera: Anthribidae), an Introduced Predator of Scale Insects in Virginia. Michael Kosztarab & Mary Rhoades, V.P.I. & S.U. 237 Annual Production and Phytoplankton Studies Within the Barrier Islands of Virginia, U.S.A. Knee land K. Nesius, Harold G. Marshall, & Charles K Rutledge, Old Dominion Univ. 251 Seasonal Changes in the Phytoplankton of Lake Chesdin, Virginia, with Ecological Observations. B. R. Woodson, Jr., Lincoln University, Pa., & Kenneth Seaburg, Xavier University, New Orleans 257 Federalism and its Impact on NSF Grants and the Training of Teachers. Ertle Thompson, Univ. of Virginia 273 NOTICES Requested Amendment to ByLaws 278 Jeffress Research Grant Awards 279 Mountain Lake Biological Station Announcement 283 Actual date of publication: February 15, 1984 Virginia Journal of Science Volume 34, Number 4 Winter, 1983 Diadophis punctatus (Linnaeus) in Virginia Charles R. Blem and Cheryl Roeding Department of Biology Academic Division Virginia Commonwealth University Richmond, Virgnia 23284 ABSTRACT Two subspecies of the ringneck snake, Diadophis punctatus , exist in Virginia. Conventional taxonomic characters supported by multivariate morphometric analyses indicate that southeastern Virginia represents the northeastern extreme of the zone of i ntergrada ti on between the southern ringneck snake, Diadophis p . punctatus , and the northern ringneck snake, p_^ e d w a r d s i i . Only the population from the extreme southeastern part of the state (Suffolk, Norfolk, and Virginia Beach) shows little evidence of intergradation. Ringneck snakes from the northern coastal plain and remainder of the state are D^ p_^ e d w a r d s i i . Individuals collected from within a triangle formed by the James River, fall line, and the southern Virginia border are intergrades. Characters of high validity for separating the subspecies in Virginia are prom inant ventral spots and neck rings. Labial/chin spotting is not useful in this regard. INTRODUCTION The ringneck snake, Diadophis punctatus, is a relatively small, secretive snake found throughout much of the eastern United States (Blanchard, 1942; Paul, 1967). Two subspecies occur in Virginia; the northern ringneck snake, IU_ e d w a r d s i i (Merrem), and the southern ringneck snake, IU_ p^ punctatus (Linnaeus). Conant (1946) described their distribution and area of intergradation on the Delmarva Peninsula and Paul (1967) examined the distribution of both subspecies in much of the rest of the southeastern United States. Conant and Paul described j populations in southeastern Virginia as D_^_ p_^ punctatus , but I; confusion still exists as to the precise range and area of f intergradation of the subspecies. Conant (1975; map 133) indicated that D^ p_^ punctatus occurs throughout much of the coastal plain, and included localities for punctatus as far north and west as Richmond. Martof et al . (1980) sTated that individuals with incomplete neck rings (= D. p. punctatus) occupy "the coastal plain of North Carolina and Virginia". A relatively large number of specimens now available permits a more precise analysis of the range limits and identification of these 208 THE VIRGINIA JOURNAL OF SCIENCE subspecies. The following study was undertaken to clarify the distribution, systematics and zone of intergradation of ringneck snakes in Virginia. MATERIALS AND METHODS Specimens (N = 172) were examined from the American Museum of Natural History (14), Carnegie Museum (49), U. S. National Museum (79), and Virginia Commonwealth University Vertebrate Collection (30). Ventral and subcaudal counts, snout-vent lengths and tail lengths were obtained from all specimens except for 1,0 that were missing part of the tail. Ventral spotting patterns, condition of the neck ring (complete or incomplete), and labial spotting were noted where appropriate. Data were compiled, sorted, and analyzed by computer. Individuals from single counties or clusters of contiguous counties were grouped subjectively and mean scale counts and frequencies of punctatus/edwardsi i characters were calculated by locality (Figs-! 1 -3 ) . Extent of geographic areas represented by each group were roughly similar. Statistical analyses were performed by the Statistical Analysis System (SAS Institute, 1982) . RESULTS AND DISCUSSION Four characters traditionally have been used to distinguish D . p . punc tatus from D^ edwards i i . Among the punc ta tus characters are: (1) chin or labial spots, (2) a neck ring interrupted mid-dorsally, (3) prom inant ventral spots, and (4) a total of ventral plus subcaudal scales of 190 or less (Blanchard, 1942; Paul, 1967; Martof et al . , 1980). Blanchard (1942) reported that ventral scale counts tend to be lower in punc tatus than in e d w a r d s i i ( punctatus males, 127-150, e d w a r d s i i males, 139-162; punctatus females, 134-155, edwardsi i females, 146-176), although the difference is not diagnostic by itself. We have used conventional methods in coordination with multivariate techniques to determine how precisely the identity of Virginia specimens might be established using all of these characters. Table 1 summarizes the frequency of occurrence of each character and the frequency of overlap of punctatus/edwardsi i criteria. Labial spots are the least reliable of the punctatus characters; of 53 individuals possessing only one punctatus character, 46 also had labial spots (Fig. 1). Snakes with only one punctatus character (ignoring labial/chin spots) were found in Mecklenburg, Surry and Henrico counties. Labial spotting occurs in specimens from throughout the state (Fig. 1), is not particularly associated with either subspecies, and therefore is not useful in identification. Individuals with two punctatus characters were obtained from Chesterfield and Southampton counties and "Nansemond" (now a part of Suffolk) and Suffolk. All seven individuals with three punctatus characters were also from southeastern Virginia (Southampton county, Suffolk, Virginia Beach). Only 9 D^ jk_ punctatus were in the sample and all were collected in extreme southeastern Virginia (Suffolk, Norfolk and "Princess Anne county" = Virginia Beach ) . The best single character for separating punctatus from edwardsi i appears to be the presence in the nominate subspecies of prominant ventral spots approximately 1/2 the width of the ventral scales. Although ventral spots occur on snakes from over a wide geographic range (91 of 172 individuals have some sort of ventral spots), prominant spots are restricted to INTERGRADATION IN VIRGINIA RINGNECK SNAKES 209 - - C CJ CD CJVr- ^ E C c c £ O CD O O -Q 4-> C ^ 03 -r- o “O ! — ^ U (U m cn GJ O'* C\J cn r— cxi e i i 03 LO OO oc co o c (O LO CX) CD CO O 2: f-H OsJ q: 1 1 > Q_ LU OsJ LT> CD CD CD C\J 1 — 1 — C\J O LO LO co r- 00 LO LO LO ^3" 00 co co o I — r— C\J cd co Q_ LU LU CL. O- LU CO CO CO O • - OsJ OsJ O LO CD CD CD O CO LO r>* 00 CO OsJ CD OsJ OsJ OsJ OsJ CO LO 00 CO CD O LO LO CD LO LO 00 OsJ 00 00 00 co o o OsJ OsJ CO I — O O OsJ OJ Cj- o Q_ LU Q_ CL LU Q- LU Q. Q- Q_ Q_ LU LU CO II -C CJ CD (J (O -C +-> lo LO r— * * cd co *3-00 r— LO OsJ CO 00 LO VO CO LO LO 00 O CD TD C C C 3 *r- 03 Q-J (/) CO OsJ CD CD CD £ CO c cd E >> 03 -Q S (O CD h- •*- CD CD +■> CJ 03 L- 03 O CD C CD CD CL E O o 10 o CL o Q. co -Q 03 Q. co 4-> O CL CO 03 S- c CD CO -Q 03 O 03 E LO CO O CL U1 CO CO fO 03 TD TD 3 3 03 03 CJ U _Q JD 3 3 CO CO + + CO CO c CD 03 S- c CD * co c CD E cj CD CL U1 CO s- CD O 03 03 CJ CO 3 03 CJ c 3 CL CO S- CD CJ 03 S~ 03 JC CJ to i- CD 0 ro S- 03 CJ CJ c 3 Cl £ “D CD CD »— C r— O Cylindrotheca closterium3 Nitzschia lengissima , Tvopedoneis lepidoptera j Ochromonas sp. and Anaoystis cyanea. Approximately 40% of the species in the Smith Island pond were also noted in the Goose Lake study. Those phytoplankters that were only recorded in the pond were the more typical estuarine and marine types indicative of saltwater intrusion. In contrast, the species recorded at Goose Lake, but not in the pond, consisted of common lake species from the Atlantic coastal plain represented by broader representation from the chlorophyceae , euglenophyceae , and a different assemblage of bacillariophyceae. The greater influence of salt water intrusion was evident in the phytoplankton composition of the Smith Island pond. In contrast to these results, Nemeth (1969) observed a variety of chlorophyceans and cyanophyceans in eight fresh water ponds during a summer survey on Delmarva Peninsula. The dominant populations included a variety of desmids, Scenedesmus spp., Tetraedron spp., and Spirogyra spp. that were not found in the Smith Island Pond. The Cyanophyceae assemblage reported by Nemeth was also different, being characterized by numerous filamentous forms (e.g., Osclllatoria spp. , Phormidlum spp.., and Anabaena spp.). In the Smith Island Pond, the dominant Cyanophyceae included Nostoc commune , Lynghya aestuajcvl, and Spirulina subsalsa. SALINE POND PHYTOPLANKTON ON SMITH ISLAND 217 Further attention is needed for the unidentified category of ceils noted in this study, which were grouped according to size (<3 ym, 3-5 ym) . Most of these appeared to be either cyanophyceans or chlorophyceans. They were generally spherically to oblong in shape, non-f lagellated , green in color, with a smooth surface. Species distinction within these cells, and within the Chlorella and Anaeystis groups is often difficult. The author questions the separation of several of these into multiple species categories and supports the re-evaluation of the marine Chlorella species {C . marina, C. salina) and the status of Nannochloris atomus as discussed by Sarokin and Carpenter (1982) . Gradations are common in this Chlorella "complex" and these forms appear to be habitat variations of Chlorella vulgaris. In addition to the phytoplankton populations in Smith pond, mention of the prominent zooplankton fauna should be given. The zooplankton was dominated by rotifers, various nauplii stages, calanoid copepods, nematodes, and ciliates, with the occasional appearance of polychete and dipteran larvae. During the fall Keratella cochlearis Brachionus sp. j Cephalodella sp., and nauplii were most abundant. In late fall and early spring, nauplii, adult calanoid copepods, and nematodes were common. During the summer Leeane sp. 3 Cephalodella sp., nauplii and nematodes were in high concentrations. The extent of the grazing influence of these zooplankton populations in determining the phytoplankton composition is unknown . In summary, the Smith Island pond was mixohaline, shallow in depth, and lacking in submerged, and emergent vegetation. A permanent standing water habitat throughout the year, the phytoplankton composition was diverse, with ultra and nanoplankton sized species most abundant. The dominant species included diatoms, a chrysophycean, and a variety of cyanophyceans. These bluegreen algae have as a common dominant several Anacystis spp. along with a variety of filamentous species. The composition was also different from assemblages reported for fresh water ponds on Delmarva Peninsula. The Virginia Barrier Islands contain a vast number of standing water habitats similar to the Smith Island pond. These sites and their biota are influenced throughout the year by overwash conditions which augment the population and assure the repeated entry of saline waters. The ponds receive drainage from the surrounding area, have a rich organic substrate, and frequently are visited by water fowl and small mammals that through defecation add to the nutrient base of the ponds. The populations that persist within these habitats are diverse, are mainly represented by small-sized species capable of a fast growth response, commonly found in high concentrations. These characterize an eutrophic condition within these pond habitats. The overwash conditions that flood these ponds will also influence the spreading of pond populations over the lower wetlands area to seed other ponds on the island, and to transport populations to the island channels and coastal waters. This innoculation and enrichment process functions in both landward and seaward directions, producing gradations of population similarities in the island ponds and coastal waters. ACKNOWLEDGEMENTS Special thanks is given to graduate assistant Leon Robert and Dr. James Matta of Old Dominion University for collecting the majority of samples used in this study and to Dr. Ray Birdsong and the Old Dominion University Barrier Island Program for providing boat access to Smith Island. LITERATURE CITED Marshall, H. G. 1980. Phytoplankton studies within the Virginia Barrier Islands. I. Seasonal study of phytoplankton in Goose Lake, Parramore Island. Va. J. Sci. 31: 61-64. 218 THE VIRGINIA JOURNAL OF SCIENCE Marshall, H. G. , K. K. Nesius, and S. J. Cibik. 1981. Phytoplankton studies within the Virginia Barrier Islands. II. Seasonal study of phytoplankton within the Barrier Island channels. Castanea. 46: 89-99. Nemeth, J. C. 1969. The summer Chlorophyceae and Cyanophyceae of the Delmarva Peninsula, Virginia. Castanea. 34; 81-86. Sarokin, D. J. and E. J. Carpenter. 1982. Ultrastructure and taxonomic obser¬ vations on marine isolates of the genus Nannochloris (Chlorophyceae) . Botanica Marina. 25: 483-491. Table 1. Phytoplankton identified in the- Smith Island pond. Presence is noted with X; the more abundant species are indicated by A, B, C, with A being the most abundant. co 4-> O O 4-> o O 00 Cr> h 4-) o o O'! bacillariqphyceae Amphiprora sp. Amphiprora alata (Ehrenberg) Kutzing Amphiprora pulchra Bailey Amphora Sp. Amphora coffeaeformis Kutzing Amphora granulata Gregory AsterioneZZa gZaoiaZis Castracane AsterioneZZa notata (Grunow) Grunow AuricuZa oompZexa (Gregory) cleve BaoiZZajoia paxiZZifer (Muller) Hendey BidduZphia sinensis Greville CaZoneis westii (Smith) Hendey Chaetooeros curvisetum cleve Chaetoceros debiZis cleve Chaetooeros sociaZe Lauder Cocooneis sp. Coscinodiscus Sp. CycZoteZZa Sp. CyoZoteZZa oaspia Grunow CyoZoteZZa gZomerata Bachman CycZoteZZa meneghiniana Kutzing CyZindrotheoa cZosterium (Ehrenberg) Reiman et Lewin CymbeZZa sp. #i CymbeZZa sp. #2 DipZoneis orabro Ehrenberg DityZum brighimieZZii (West) Grunow Eunotia praerupta Ehrenberg FragiZaria sp. Gyro sigma Sp. Gyrosigma baZticum (Ehrenberg) Cleve Gyrosigma wansbeokii (Donkin) Cl eve Hantzsohia amphioxys (Ehrenberg) Grunow X - - X - - - X - - X X X X - - X - - - X X X X X X V - X X X X X X v - - A - X - - - X A X X c X X X X - - - - - X - X X A _ _ _ _ _ _ _ _ _ - X - X v X X c A X - c - X - A X - - - - - - X - c - B - - X - - - - A X X c - X A B - - - X X X X c X X X X B X X v : : _ X A - X - - - X X X X X - X - - - - - X - v - - X X X - - - ” A _ _ _ _ - - - - SALINE POND PHYTOPLANKTON ON SMITH ISLAND 219 Table 1. (continued) Leptooylindrus danious Cleve Li omophor a sp. Liomophora paradoxa (Lyngbye) Agardh Melosira distans (Ehrenberg) Kutzing Melosira moniliformis (Muller) Agardh Melosira nummuloides (Dillwyn) Agardh Navioula sp. #1 Navioula sp. #2 Navioula arvensis Hustedt Navioula spioula (Dickie) Clebe Nitzsohia sp. Nitzsohia olausii Hantzsch Nitzsohia longissima (Brebisson) Ralfs Nitzsohia paradoxa (Gmelin) Grun Nitzsohia sigma (Kutzing) Smith Nitzsohia vermioularis (Kutzing) Hantzsch Rinnularia sp. Rinnularia major (Kutzing) Rabenhorst Plagiogramma sp. Rhizosolenia alata Brightwell Rhizosolenia delioatula Cleve Skeletonema costatum (Greville) Cleve Stenopterohia anoeps (Lewis) Brebisson Stephanopyxis palmeriana (Greville) Grunow Surirella sp. Surirella fastuosa Ehrenberg Surirella striatula Turpin Synedra sp. Thalassionema nitzsohioides Hustedt Thalassiosira sp. Tropedoneis lepidoptera (Gregory) Cleve Unidentified centrales Unidentified pennales CRYPTOPHYCEAE Chroomonas sp . Chroomonas oaroliniana Campbell Cryptomonas sp. #1 Cryptomonas sp. #2 CHLORQPHYCEAE Chlamy domonas sp. Chlorella sp. Chlorella vulgaris Beyerinck Cladophora sp. 00 CO -p > u c tH -p o 0 M a i 3 3 Cn CD -P o o 2 3 < i-o 3 CO O o CO CM S cO O < CO O r- CM O'! i — i CM pH r- r-l CTi - X - X y - - - - - X X - - A X X - - - - - X X - - - X X - - X X X X X c X - X - - - - c - X 7\ - - X - - - X X X X X - - - X - X c B X - X X X X - - X - X X - - - - - - X X - X X X - X - X - X - X - - - X - - - - - - - X _ _ _ _ _ X - - - - - X X X - - - - - - - - X - - - - - - - - - - - - - - X - - X X - - X - - - - - - - X X X - X X X - X X _ _ - _ - - X - X X X - - X X - - X X X X A - - - - - A X X X c - X - - - X - - - - - - - - - - X - - - - X - - - - - - - X X - - - - X X X X - X - X - X - X X X X - - - - - c X A - 220 THE VIRGINIA JOURNAL OF SCIENCE Table 1. (continued) 00 +j -p o > o CD p p a. c p P ty ft CD •p o o s (T3 < T> t> p c n u o r-~ CO 1 — 1 CM CM £ CTI I — ( CO CM o 1 — 1 Cladophora glomerata Kutzing _ _ _ _ _ - - - X - Nannochloris atomus Butcher X c - - - - - X X X Oedogonium sp. #1 X X - X X - - - X - Oedogonium sp. #2 X Scenedesmus sp. #1 - - X - X - - - - - Scenedesmus sp. #2 — — ' " X CHRYSOPHYCEAE Chrysococcus minutus (Fritsch) Nygaard - - - - X - - - - - Ochromonas sp . X - c X c X X X X X Ochromonas minis cula Conrad X X Ochromonas variabilis Meyer X — “ X cyanophyceae Agmenellum quadriduplicatum (Meneghini) - - X - - - - - - - Brebisson Anabaena confervoides Reinsch - - - - - - - X - - Anabaena laxa Rabenhorst - - X - - - - - - - Anacystis aeruginosa Drouet et Daily c - X - c - - X X X Anacystis cyccnea (Kutzing) X X X - X - X X A X Drouet et Daily Anacystis dimidiata (Kutzing) X X - - - - - - X X Drouet et Daily Anacystis marina (Hansgrig) X X - - - - X X X X Drouet et Daily Aphanizomenon flos-aquae (L.) Ralfs X — Calothrix aeruginea (Kutzing) Thuret X - Coccochloris elabens Drouet et Daily - - - - X - - - - - Gomphosphaeria aponina Kutzing X - - - - X X - - X Johannesbaptistia pellucida (Dickie) X X - c X - X - - X Taylor et Drouet Lyngbya aestuavcii (Mert.) Liebmann - - - X - - B - X - Lyngbya confervoides Agardh X X - - - - - - - - Lyngbya semiplena Agardh - - - X - - - - - - Microcoleus vaginatus (Vaucher) Gomont - X X - - - — “ - - Nodularia sp. - - - X - - - - - - Nodularia spumigena Mertens X - - X c - X - - - Nos toe sp. X - 'Nos toe commune Vaucher X - - - X X B “ X - Oscillatoria sp. #1 X X - X X - - - - X Oscillatoria sp. #2 X X - - - - - - - X Oscillatoria curviceps Agardh - X Oscillatoria erythraea (Ehrenberg) - X - - - - “ - - Kutzing Oscillatoria omata Kutzing - - - X - - - - - “ Oscillatoria principes Vaucher - - - - - - - - X c Oscillatoria submembranacea Ardissone X X - X X - X - X X et Strafforella SALINE POND PHYTOPLANKTON ON SMITH ISLAND Table 1. (continued) 221 CO +J o o [■" -P O o 00 tn -POO 0 0 2 O co oo F' fH CN CN CO P c p a p P C O 2 i — I CN tn P < r" Tetraselmis maculata (Kylin) Butcher - X Tetrasebmis striata Butcher B X X XANTHOPHYCEAE Stipitocooous sp. Tribonema sp. Tribonema minus (Willie) Hazen Tribonema angustissimum Pascher Tribonema taeniatum Pascher Unidentified cells <3 yin Unidentified cells 3-5 ym X-------- -------X- -X-X-X-XX XX-XX-XXXX X--X-XXX-X 18 Sep Virginia Journal of Science Volume 34, Number 4 Winter, 1983 / f- Introduction oi&nthribus nebulosus (Coleoptera: Anthribidae) in Virginia for Control of Scale Insects: A Review Michael Kosztarab and Ferenc K< Lozar Department of Entomology Virginia Polytechnic Institute and State University Blacksburg, VA 24061 and Hungarian Research Institute of Plant Protection Budapest, Hungary, H-1525 respectively. ABSTRACT A new biological control agent, Anthribus nebulosus Forster, was introduced from Hungary to Virginia during 1975 and 1977, successfully field-released in 1978, and established in the state by 1981. Methods of collection, introduction, rearing and release of the beetles are presented. A comparison was made of climatic conditions in its native habitat with that of the release site. Known distribution, life cycle, feeding habits, the scale insect prey, the natural enemies of A. nebulosus and other associated organisms are reviewed, as are reports on its effectiveness as a biological control agent in Europe. This was the first introduction of this beneficial beetle to North America. Anthribus nebulosus feeds on and destroys colonies of scale insects of about 15 species in Europe and Central Asia. Four of these scale insects are also pests in the Eastern United States. These include the European fruit lecanium, Parthenolecanium corni (Bouche); nut scale*, Eulecanium tiliae (Lin.); and the small and the large spruce bud scale*, Physokermes hemic ryphus' Dalman, P. piceae Schrank. Because the beetles feed on closely related scale insects in Europe, they have the potential to also control the cottony maple scale, P. innumerabilis (Rathvon); cottony peach scale, Pulvinaria amygdali Cockerell; cottony vine scale, P. viti s (Lin.); the European peach scale, Parthenolecanium persicae (Fabr.); Fletcher scale, P. f letcheri (Cockerell); oak lecanium, Parthenolecanium quercifex (Fitch), and terrapin scale, Mesolecanium nigrof asciatum (Pergande) in North America. They are known to destroy up to 90% of certain scale insects in their native land. Within three years after release, we have achieved over 30% *These are common names from European literature. 224 THE VIRGINIA JOURNAL OF SCIENCE control at a release site in southwest Virginia. Beetles had dispersed as far as 13 km by Fall 1982. INTRODUCTION There are a number of soft scales in the Eastern United States without effective natural control agents. Many of the same species are also pests in Europe, but their native biological control agents usually keep the pest populations under economic injury level. Anthribus nebulosus Forster, is an effective predator for at least 15 species of scale insects in Europe and Central Asia (See Table 1). The beetle belongs to the family of fungus weevils, Anthribidae. The adults are somewhat robust, convex, and oval, about 2.0 - 4.0 mm long, and 1.5 - 2.0 mm wide, and grayish-black. In 1975 work was initiated for the evaluation, collection in Europe, introduction to the United States, rearing and release of this new biological control agent. The following is a review of the available information on this insect and on our work completed between 1975 and 1982. DISTRIBUTION AND CLIMATIC FACTORS Anthribus nebulosus is considered an Euro-Siberian species with distribution in Central Palearctic, in areas of deciduous forests, spruce forests, orchards and on ornamental trees when these are infested with their prey scale insects. Prior to 1978, it was reported from the following countries: Bulgaria, Finland, France, Germany, Hungary, Poland, and USSR (Ukraine to Eastern Kazakhstan) . It is more common in Central Europe than the related species, A. f asci atus Forster, which has a more southern distribution and was reported as far south as Italy by Silvestri (1919). Prell (1925) reported A. nebulosus to be more common in Germany than A. f asciatus . Anthribus nebulosus prefers the temperate continental climate with moderately low humidity and precipitation. To obtain some indication of their chances for survival in SW Virginia, we compared the temperature records for a six year period (July 1976 - June 1981) from their area of collection at Csopak, Hungary, (Fig. 1), with that of Blacksburg, Virginia (Fig. 2). The comparison shows similarities in monthly averages, but there are more frequent extremes in lower minimum and higher maximum temperatures in Hungary, when compared with the same monthly averages from Blacksburg. There was a higher amount of precipitation in Blacksburg (Fig. 3). The relative humidity was higher from April through September, and varied less in Blacksburg, while in Csopak, Hungary, the monthly relative humidity averages were higher from October through December (Fig. 3) . LIFE CYCLE AND PREY SCALE INSECTS The life cycle of the beetle is intimately related to the phenology of the host scale species. Anthribus nebulosus and all 15 species of scale insects (Table 1) that serve as host for the beetle have one yearly generation in the areas studied. Matesova (1966) studied the biology of Arthribus nebulosus in the dry steppe regions of Eastern Kazakhstan (USSR). Illes et al (1896) in Hungary, and Goanca et al (1974) in Moldavia (USSR), studied the same beetles under the moderately dry continental climate of east central Europe. Schmutterer (1952) and Forster INTRODUCTION OF ANTHRIBUS NEBULOSUS IN VIRGINIA 225 Table 1. Known host scale insect species preyed on by Anthribus nebulosus Forster Species of Coccoidea Fam. Coccidae (soft scales) Eulecanium caraganae Borchsenius Eulecanium ci liatum (Douglas) Eulecanium douglasi Sulc Eulecanium sericeum (Lindinger) Eulecanium ti liae (Lin.) Nemolecanium abieti s Borchsenius Parthenolecanium corni (Bouche) Parthenolecanium rufulum ( Cockerell ) Physokermes hemic ryphus Dalman Physokermes inopinatus Danzig & Kozar Physokermes piceae Schrank Pulvinaria betulae (Lin.) Rhodococcus perornatus (Ckll. & Parrott ) Rhodococcus spireae Borchsenius Known Areas of Predation* USSR: Leningrad Region (3) S. Germany (18,19) USSR: East Kazakhstan (14) S. Germany, Poland (10,18) France, S. Germany, USSR: Leningrad Region, E. Kazakhstan (3,7,14,15,18,20) USSR: Ukraine (23) As above (E. tiliae ) , Hungary (3,7,9,14,18,20,23) S. Germany (18,20) Germany, USA: Virginia (since 1978 ) ( 5 , 7 , 1 1 , 19 , 20 ) Hungary (12) Germany, USSR: Leningrad Region, E. Kazakhstan (3,5, 11, 14, 19,20) USSR: Ukraine (23) Bulgaria (24) USSR: E. Kazakhstan (14) Fam. Kermesidae (gall-like scales) Kermes quercus (Lin.) Germany (15) Host scale undetermined, probably Physokermes piceae Finland (17) *The numbers in parentheses correspond to articles listed in the References Cited. 226 THE VIRGINIA JOURNAL OF SCIENCE INTRODUCTION OF ANTHRIBUS NEBULOSUS IN VIRGINIA 227 228 THE VIRGINIA JOURNAL OF SCIENCE (1973) studied them under the more humid conditions of west central Europe. Climatically, our area of release in southwest Virginia is closest to west central Europe. Our comments below are based on a synthesis of all these works plus our own. In both Moldavia and southwest Virginia overwintered adults appeared for the first time from mid-April; fed on scale honeydew, all stages of scale insects, and on fungi; laid eggs from early May through early June. These were deposited by chewing a hole on the still soft body side of the scale insect female and laying an egg in the brood chamber and by plugging the wound with their salivary secretion. These egg-laying sites later showed as dark spots on the hardened scale body. Egg laying of A. nebulosus was often successfully prevented by the attack of ants in Germany. The larvae developed and pupated under the scale exoskeletons from mid-June through July, the pupal period lasting from one to a few weeks; adults emerged by chewing a hole on the lateral side or on dorsum of the female scale exoskeleton; newly emerged adults fed on remains of old scale insect female exoskeletons ("mummies"), on scale insect honeydew and on remains of scale insect eggs in Europe and in Virginia. The beetles increased to high populations only during the year following the scale insect outbreaks in western Europe. This was also our own conclusion at Csopak (Fig. 4). The adult beetles went into diapause as early as August in S. Germany, and August and September in southwest Virginia. The diapause lasted until the following spring. The beetles chose leaf litter and crevices in the trunk and branches for overwintering in Moldavia (Goanca et al 1974), but in Finland and Germany (Schmutterer 1952, 1965; Forster 1973) each beetle chewed a small gallery in the bark and rested at its end. For shelter they used bark crevices and empty female scale insect exoskeletons in Germany (Forster 1973) and in southwest Virginia (Kosztarab & Rhoades 1983) and leaf litter and scale exoskeletons in Hungary (Illes et al 1896). The available literature agrees that larvae feed individually or rarely in groups of 2 or 3 under each egg laying scale insect female, and devour the eggs as they are being laid. NATURAL ENEMIES OF ANTHRIBUS NEBULOSUS AND SOME ASSOCIATED ORGANISMS Among the natural enemies of adult beetles in Central Europe, Csiki (1906) listed five species of titmice: Parus ater Lin., P. coeruleus Lin., P. ma j or Lin., P. palustris Lin., and Aegithalus caudatus Lin. Stomach content studies of these birds proved that they fed on beetles from October through February (1901-1904). There were 4 to 28 beetles per bird stomach during March-April and during August-September . This number increased to between 6 and 50 in December. During December a total of 267 beetles occurred in 9 titmouse stomachs. It was assumed by Csiki that, because of snow cover in December, the birds were forced to feed in the tree crowns where they found overwintering beetles inside shelters provided by bark crevices and the dead scale insect exoskeletons. The snow cover is normally thicker and lasts longer in Hungary than in the Mid-Atlantic states. This is why we believe that birds will not significantly affect the beetle populations in the area of our releases. Organisms found on the host trees, Norway spruce, ( Picea abies (L.) Karst), associated with A. nebulosus at the collecting site in Csopak, Hungary, between 1975 and 1981 were: Dermaptera: Forf iculidae : Forf icula auricularia Lin., European Earwig. Since its population size followed the scale population increase it is probably a predator of the scale host. Fig. 4. Ten Year Population Fluctuation of Scale Host, Physokermes inopinatus (solid line) and Predation Rate by Anthribus nebulosus (dash line) at Csopak Hungary. INTRODUCTION OF ANTHRIBUS NEBULOSUS IN VIRGINIA 229 snqxxqquy Aq uoxqnpaxj qo % *Rate of scale infestation is given in a scale of 0 to 4, after Borchsenius (1950) as follows: 0=scales absent; l=only few single individuals present; 2=scales form small colonies of 2 to 10; 3=scales form large colonies of 11-100; 4=plants generally infested. 230 THE VIRGINIA JOURNAL OF SCIENCE Hemiptera: Lygaeidae: Gastrodes grossipes (DeGeer) . It is known only from bark and cones of conifers. Neuroptera: Chrysopidae: Chrysopa perla Lin. Larvae are predators on a variety of insects. Coleoptera: Coccinellidae : Adalia bipunctata (Lin.), Coccinella septempunctata Lin., Exochomus quadripustulatus (Lin.) and Scymnus ( Pul lus ) impexus Mulsant. All four lady beetles are known predators of scale insects. Bruchidae: Bruchidius nudus Allard. This seed weevil is known from Cyti sus triflorus L'Herit ( Leguminosae ) only. Hymenoptera: Eurytomidae: Eurytoma sp. was found as an external parasite of beetle pupae in the material collected by the authors at Csopak, Hungary, during June 1981. In addition, honeydew collecting ants in the genera Camponotus and Formica , also members of the species and ants Lasius niger Lin., L. alienus Forster attacked the beetles and often successfully prevented their egg laying on host scale insects (Schmutterer 1952, 1965). PROCEDURE FOR INTRODUCTION TO VIRGINIA Collecting in Europe : The adult beetles have been collected each year (except in 1976) since 1975 at Csopak, Hungary, from Norway spruce trees that were infested with Physokermes inopinatus scale insects. The trees were 18-26 years old, with an average height of 8 m. Usually 12 to 16 of the most heavily infested trees were selected for collecting. Best results were obtained during sunny days without wind, around 24°C, between 11 AM and 2 PM, from end of June through early July. White cloths were placed on the ground under the tree and the branches starting from the top were vigorously beaten and shaken with a long handled garden tool to dislodge the beetles into the cloths. Contents from the cloths were shaken into a wide mouth glass jar and/or into a large plastic bag when it contained too much residue from the- tree. Because most of the beetles hold on to the cloths, most of the tree residue could be easily separated from the beetles with light shaking of the cloths. Only very few beetles flew away. The collected and separated beetles were supplied with fresh branches of spruce with live scale first instars, adult female scale exoskeletons, and with water in cotton stoppered vials. Each tree provided from 18 to 90 beetles in 1981. The tree residue that was collected in large transparent plastic bags contained small broken twigs, needles, scale insect exoskeletons that often included live adult beetles or their pupae, different parasites and predators of the beetles and/or of their host scale insect, and other associated organisms. This residue provided many additional beetles when the plastic bag containing the residue was placed in the sun with its closed top directed up. The plastic bag was utilized as a sun-heated inverted Berlese funnel. The beetles and some other live arthropods, due to the heating of the contents of the plastic bag by the sun, aggregated on top, near the closed top of the bag, where they were collected at regular intervals. It was practical to check these bags during the day from 10 AM to 4 PM every half an hour. We removed the beetles and other organisms from the top and transferred them into rearing containers. A 700 gr residue INTRODUCTION OF ANTHRIBUS NEBULOSUS IN VIRGINIA 231 Fig. 5 Plastic box with screened top and corrugated paper rolls used for shipping beetles to the United States; also used for overwintering. sample provided 206 beetles from 10 to 4 PM on July 10, 1981 (first day after field collecting). Shipping . Beetles were kept with food (live and dead scale insects, sooty mold fungi on needles) and water in wide mouthed rearing jars (2 1.) covered with cheese cloth, until ready for shipping. Rolls (4 to 6) of corrugated paper (as in Fig. 5) were included in each jar for shelter. About 80% of beetles displayed positive thigmotaxis, by moving into the grooves of the corrugated paper rolls, and a few inside exoskeletons of scale insects. They became motionless in these shelters and went into summer diapause. Just before shipping, the beetles were transferred into rigid plastic containers (25 X 17.5 X 6 cm) with screened tops for ventilation (Fig. 5). The copper screen had 600 y square mesh openings to prevent escape. The water vials were attached to the bottom of the container to avoid rolling while in transport. The plastic containers with the beetles were brought with personal luggage on regular aircraft to the United States. Rearing . After arrival in Blacksburg, Virginia, the beetles were transferred into cages with wood frames and organdy walls, a bottom of 1/2" plywood and a sliding door of thick window glass. The beetles were provided with shelter of corrugated paper rolls, and for food with dead adult females and live nymphs of Parthenolecanium quercif ex and Physokermes hemicryphus . The scales were on twigs with leaves of pin oak, Quercus palustris Muenchh. , or needles of Norway spruce placed in water-filled 250 ml. beakers with cotton plugs. Water was provided in cotton stoppered vials. The food sources and water were replenished as needed once or twice weekly. The beetles were kept under observation for possible parasitization and for pathogens carried 232 THE VIRGINIA JOURNAL OF SCIENCE with them from the country of origin. Only in one case did we find a parasitic hymenopteran in Hungary before shipping. After feeding and drinking, many of the beetles retreated into the shelters for summer diapause that often continued without interruption into winter diapause. During November, the corrugated paper roll shelters with the beetles were transferred for overwintering into securely locked double cages. Overwintering . Food and additional shelters were provided with the inclusion of scale insect exoskeletons, in the overwintering boxes, a 5 or 10% sugar solution, and tap water, which were added in cotton or sponge stoppered vials. For overwintering we used the same plastic boxes with screen tops (Fig. 5) that were used earlier for their transportation. The box was placed in a 28 1. plastic trash can filled with dry oak leaves and securely covered with organdy. The plastic can was placed in the closed rearing cage, provided with a polyethylene roof, and the latter in a large locked, screen walled cage. We found the overwintering mortality to be 32-67% when the beetles were overwintered under outdoor conditions in Blacksburg during 1977 and 1981. Our earlier attempts during the winter of 1975-76 to keep the overwintering beetles inside growth chambers, simulating outdoor winter conditions, were not successful. In this experiment the beetles were kept inside wide mouth 3 1. jars covered with cheese cloth and were provided with honey-water, and distilled water in cotton stoppered vials, spruce shoots with P. hemicryphus , and oak shoots with Kermes kingi Cockerell nymphs and dead adults. Water and food were checked and replenished weekly as needed. Day-night photophases were synchronized with the shorter day length during the winter months, simulating the outdoor conditions. During the first week of April, the overwintered beetles were transferred into rearing cages (described earlier) and kept inside the laboratory, where potted oaks infested with Kermes kingi and Parthenolecanium quercif ex provided food and egg laying sites for the beetles. Establishment in Virginia . After three years (1975-1978) of rearing experiments, the progeny of our 1977 introduction (ca 150 beetles) were field released during October 1978 in Blacksburg, SW Virginia (VPI & SU campus), on Norway spruce trees infested with Physokermes hemicryphus live nymphs and dead adult female exoskeletons. The, beetles were individually placed with camel's hair brush on twigs infested with this scale insect. The procedure was repeated with ca 200 more beetles on the same trees during September 1979. Monitoring of Released Populations . Scale insect exoskeleton samples were taken and the trees checked three times yearly, starting during the spring of 1979, and continuing each summer, fall and spring through 1982. The predation rate reached 30.1% by August 1981 (Fig. 6). After 1980, we ptarted checking the surrounding area for evidence of beetle dispersal. The beetles were recaptured in Blacksburg, at about 1 km east of the original release site during Fall 1981 and Spring 1982. During the Fall 1982 a few beetles were captured on trees at about 13 km direct line distance southwest (Radford University) of the release site. At each location the beetles fed on P. hemicryphus present on Norway spruce trees. To provide evidence of dispersal and predation, we utilized a light colored vinyl-nylon tarpaulin of about 4 X 5 m. size. The tarpaulin was placed on the ground before vigorously shaking INTRODUCTION OF ANTHRIBUS NEBULOSUS IN VIRGINIA 233 the spruce branches. The fallen residue was hurriedly shaken into plastic bags to prevent escape of beetles, and it was checked in our laboratory under a dissecting microscope for beetles and/or evidence of predation (emergence holes on collected scale insect exoskeletons). This procedure repeated during the months of April, June or July and Septem In addition to the large ragged beetle emergence holes, we /found large numbers of small circular chalcid parasite emergence/holes. Field Release at Virginia Beach. During Fall 1981, we field- released about 300 more beetles at a new site at Virginia Beach on pin oak trees infested with oak lecanium, Pi quercif ex (Schultz 1981). Our Spring 1982 recapture saddles did not provide evidence of their survival and/or 'establishment in Eastern Virginia. Due to budgetary limitations, we had to temporarily discontinue further biological control work in that area . RATE OF PREDATION BY ANTHRIBUS NEBULOSUS Geza Horvath, as given in Illes et al (1896) reported from Hungary that among the three most effective natural enemies of the brown scale, Partheno lecanium corni , in black locust forests, ( Anthribus nebulosus , the chalcid, Coccophagus scutellaris Nees, and the lady beetle, Exochomus quadripustulatus Lin.), A. nebulosus ( ^various ) was the most effective. It was so well established in certain areas in 1890, that the reduction of scale populations was remarkable. Anthribus nebulosus was found by Dyadechko (1954) to significantly reduce the number of soft scales on oaks in the forests of the Ukraine. A similar observation was made by Mate.sova (1966) in the forests and steppe zones of Eastern Kazakhstan, and by Danzig (1959) in the Leningrad Region. The latter author noted that ca 30% of Eulecanium caraganae eggs, and ca 50% of Partheno lecanium corni eggs were destroyed by the larvae of this beetle. Fig, 6. Predation of Anthribus. nebulosus Forst 234 THE VIRGINIA JOURNAL OF SCIENCE Klausnitzer and Forster (1976) found A. nebulosus to be the dominant natural enemy of large spruce bud scales, Physokermes piceae , in the Dresden area, Germany, and it was responsible in 1971 for 58.6%, and in 1972 for 37.9% of the scale insects destroyed. Schmutterer (1965) also reported A. nebulosus as often destroying 90% of P. piceae colonies in Germany. Predation by this beetle on Norway spruce infesting Physokermes inopinatus at Csopak, Hungary, from 1973, through 1982 ranged from 3 to 55% (Fig. 4). Three years following release, the beetles on our observation trees in Blacksburg, Virginia (Fig. 6), destroyed 30.1% of the Norway spruce infesting Physokermes hemic ryphus . NEEDED FUTURE WORK AND PLANS Future studies on this species should include an investigation into economical mass rearing methods and a determination of its ecological requirements, including optimal indoor overwintering conditions. After a suitable host infestation is found, the beetles should be released in the Central States, eg. Kansas and Oklahoma, where the climatic conditions are similar to its natural range in Europe and Central Asia . Multiple introduction of foreign parasites and predators of pest scale insects was strongly recommended by Rosen (1973) on both theoretical and practical grounds. New release attempts in the Norfolk area of Virginia where a number of nurseries occur might help to control oak lecaniums and other scales causing heavy damage on oaks and other ornamental and shade trees. New introductions, and further rearings are planned at VPI & SU, and as soon as funds are made available, the beetles will be released in other parts of Virginia, especially in the major fruit growing areas, such as the Shenandoah Valley. ACKNOWLEDGEMENTS The authors express thanks to the following persons who assisted their work. Help with collecting of beetles in Hungary was given in 1977 and 1978 by Dr. and Mrs. John J. Drea of the USDA European Parasite Laboratory, and in 1977 by Mrs. Galina M. Konstantinova of the Plant Quarantine Laboratory, Moscow. Identification of associated organisms of this predator was provided by USDA Systematic Entomology Laboratory scientists: Drs. R. D. Gordon ( Coccinel lidae ) , I. E. Grissell ( Eurytomidae ) , T. J. Henry (Lygaeidae) and F. M. Kinsolver (Bruchidae); also by the Hungarian Research Institute of Plant Protection scientist Dr. F. Szentkiraly ( Chrysopidae ) . Assistance with the release of beetles at Virginia Beach was given by Drs. T. J. Banko and P. B. Schultz, both Research Scientists at the Virginia Truck and Ornamental Research Station (VTORS) at Virginia Beach, Virginia. From the Virginia Polytechnic Institute (VPI & SU) : Ms. Mary Rhoades assisted with the computation of climatic records and with the preparation of this article; Dr. L. T. Kok, Professor, critically reviewed the first draft of this manuscript and made a number of suggestions; weather records for Blacksburg were provided by Ms. Ellen L. Howard, Statistician (VPI & SU) . The weather records for Csopak were given by Dr. Kalman Sieto (Director of Plant Protection and Agrochemicals Station of Veszprem County at Csopak), who also gave permission since 1975 for collecting beetles on the premises of the station. Six Camellia plants were donated for rearing soft scales by Mr. E. C. INTRODUCTION OF ANTHRIBUS NEBULOSUS IN VIRGINIA 235 Hogge of Chesapeake, Virginia; and four Ilex sp . plants were also given by Dr. E. A. Borchers, Director of VTORS . Funds for partial cost of the 1979 collecting trip were provided through a National Science Foundation grant ( NSF-323913 1 ) . This research project was supported by the Virginia Agricultural Experiment Station (Project 2024390). REFERENCES CITED 1. Borchsenius, N. S. 1950. Collection and study of armored and soft scales (In Russ.). Akad. Nauk Zool. Inst. Zhivotnyi Mir SSSR 4:30 p. 2. Csiki, E. 1906. Proven records on feeding habits of our birds. (In Hungarian). Aquila (Budapest) 13:148-161. 3. Danzig, E. M. 1959. The scale insect fauna (Homoptera, Coccoidea) of the Leningrad region. Entomol. Review 38:395-407 . 4. Dyadechko, N. P. 1954. Predators and parasites of scale insects on oaks in Ukrainian SSR (In Russ.) In Biological Control of Pest Insects. Akad. Nauk Ukr . SSR 5:129-135. 5. Forster, G. 1973. Zur Biologie und Morphologie von Anthribus nebulosus Forster (Col., Anthr . ) . Entomol. Nachr. 17:117-121. 6. Goanca, I. K., E. S. Sugon j aev and E. M. Danzig. 1974. Scale insects and soft scales of Moldavia and their natural enemies. (In Russ.) Izd. Kartya Moldov. Kisinev, 112 p. 7. Hoffman, A. 1945. Coleopteres Bruchides et Anthribides. In Faune de France, No. 44. Paris, P. Lechevalier. Spec. Famille des Anthribidae, pp . 124-159. 8. Illes, N. , J. Havas, G. Horva/th and J. Vadas. 1896. The Black Locust Scale problem in Hungary (In Hung.). National Forestry Assoc., Budapest, 1896. 207 p. 9. Jablonowski, J. 1916. The scale insects of grapes and other economic plants (In Hung.). Kiserletugyi Kozl. 19(2) : 1-120. 10. Kawecki , Z. 1955. Studies on the genus Lecanium Burm. Part III. Lecanium sericeum Ldgr . < (Homoptera, Lecaniidae). English transl. 1-10 p. OTS 60-21405, U.S. Dept. Comm., Washington, DC) Reprinted from Polska Akad. Nauk Inst. Zool. Ann. Zool. 25 ( 24 ) : 2 13 -22 6 . 11. Klausnitzer, B. and G. Forster. 1976. Zur Kenntnis der Parasitierung ; weiblicher "Brutblasen" der Fichtenquirlschi ldlause Physokermes piceae Schrk. und Physokermes hemic ryphus Dalm. Hercynia N. F. , Leipzig 13(1) : 85-92 . 12. Kosztarab, M. and F. Kozar. 1978. Scale insecrs--Coccoidea (In Hung.). Fauna Hungariae No. 131, Vol. 17 (22): 1-192. 13. Kosztarab, M. and M. Rhoades. 1983. Food consumption, mating behavior and shelter selection of Anthribus nebulosus Forster (Coleoptera: Anthribidae), an introduced predator of scale insects in Virginia. Va. J. Sci . 34(4). 236 THE VIRGINIA JOURNAL OF SCIENCE 14. Matesova, G. Ya. 1966. Beetles of the genus Brachytarsus (Coleoptera, Anthribidae) that: are enemies of soft scales (Homoptera, Coccoidea) in Eastern Kazakhstan. Entom. Rev. 45(2 ) : 141-142 . 15. Prell, H. 1925. Beitrage zur Biologie des grauen Schi ldlausrusslers (Anthribus nebulosus Forst. ) . Z. Forst- u. Jagdwesen 57:245-250. 16. Rosen. D. 1973. Methodology for biological control of armored scale insects. Phytoparasi tica l(l):47-54. 17. Saalas, U. 1917-1923. Die Fichten-Kaf er Finnlands, Parts I & II. Suom. Tiedeak . A8(1917)/ A22(1923): 80. Porvoo, Helsinki . 18. Schmutterer, H. 1952. Die Okoiogie der Cocciden (Homoptera, Coccoidea) Frankens (pt. 3). Ztschr. f. Angew. Entomol. 33:65-100. 19. Schmutterer, H. 1965. Zur Okoiogie und wi rtschaf tlichen Bedeutung der Physokermes Arten (Homopt. Coccoidea) an Fichte in Siiddeutschland . Ztschr. f. Agnew. Entomol. 56(4) : 300-325 . 20. Schmutterer, H. 1972. Unterordnung Coccoidea, Schildlause. Coccidae ( Lecaniidae ) , Napf schi Id-Soft scales. Ua Die Forstschadlinge Europas. Ein Handbuch in funf Banden, by W. Schwenke . p. 405-418. 21. Schultz, P. B. 1981. Release of a scale insect predator in Tidewater. Virginia Ornamentali st 8(5): 7. 22. Silvestri, F. 1919. Contribuzioni alia conoscenza degli insetti dannosi e dei loro simbionti. V. La Cocciniglia del Nocciuolo ( Eulecanium coryli L. ) Portici R. Scuola Super, di Agr . Lab. Zool. Gen. e Agr. Bol. 13:127-192. 23. Tereznikova, E. M. 1981. Fauna of Ukraine. Vol. 20. Scale Insects. (In Ukrainian) Part 19. Zool. Inst., Acad. Ukr . SSR . 215 p. Tsalev, M. 1966. Contribution to the morphology, biology and control of Rhodococcus bulgariensis Wunn (In Bulgarian) (Homoptera, Coccoidea) in Bulgaria. Plant Science 3(7):3-14. 24. Virginia Journal of Science Volume 34, Number 4 Winter, 1983 / Food Consumption, Mating Behavior, and Shelter Selection of Anthribus nebulosus Forster (Coleoptera: Antfiribidae), an Introduced Predator of Scale Insects in Virginia Michael Kosztarab and Mary Rhoades Department of Entomology Virginia Polytechnic Institute and State University Blacksburg, Virginia 24061 ABSTRACT Anthribus nebulosus Forster adult beetles, whose larvae are predators of scale insect eggs, were observed in the laboratory from late July to mid-November and again after overwintering from mid-April to mid-August to study their feeding habits, shelter preferences, and mating behavior. Natural honeydew from 5 species of scale insects, artificial honeydew, sugar-water solution, water and sooty mold on twigs and leaves were consumed. All stages of live scale insects were eaten by beetles in the spring, but not in the fall. Dry exoskeletons of old female scale insects were used as food both times of the year. Response to shelters, which was studied only in the fall, was variable; 25% of the beetles eventually entered a shelter and stayed there. Old female scale insect exoskeletons were preferred by the beetles to black paper shelters. The average weight of a sample of beetles in mid-November was 3.6 mg. Length of 20 beetles averaged 4.0 mm (range 3. 0-4. 8), and width 1.7 mm (range 1.2-2 . 1) . Mating was recorded from April 14 to May 3, with copulation lasting 3 to 35 minutes. The pr'eoviposi tion period after mating was 1-2 weeks, and the egg incubation period about 4 days. The combined larval and pupal stages lasted about 4 weeks. Adult longevity, after overwintering, ranged from 2 weeks to 4 months with males living as long as females. INTRODUCTION Anthribus nebulosus (Fig. 1) is an important predator of soft scale insects in Eurasia (Kosztarab & Koz^r, 1983). Some of its prey scale insects (Family Coccidae) are also pests in North America, such as the European fruit lecanium, Parthenolecanium / - corni (Bouche); the nut scale*, Eulecanium tiliae (Linn.); and the small spruce scale*, Physokermes hemic ryphus (Dalman) . But *European common English names were used for these species because there are no approved common names in North America. 238 THE VIRGINIA JOURNAL OF SCIENCE some of their Eurasian predators are absent from the American fauna. Although the first author released A. nebulosus in Virginia in 1978 and they have become established in low numbers in three localities in southwest Virginia, little was known about their biology except the general life cycle (Kosztarab & Kozar 1983). Matesova (1966) stated that there is only very limited information on its biology. Our search for literature on this species provided about 26 articles, but most of these gave little useful information, and what was available was often contradictory. Our objectives with these laboratory studies were to find out their feeding habits, shelter preferences, mating behavior and development. MATERIALS AND METHODS Fall Observations Adult beetles were collected at Csopak, Hungary, on July 8, 1981, from Norway spruce trees Picea abies (L.) Karst, infested with Physokermes inopinatus Danzig and Kozar and maintained in the VPI&SU Beneficial Insect Introduction Laboratory until their use for these tests. All laboratory observations were made at 17-33°C, relative humidity 30-90%. One or more beetles were placed in plastic or glass petri dishes with the various materials to be tested for their attractiveness to the beetles (Fig. 2), and observed for several weeks. Two sizes of dishes were used, 9 cm wide x 1.9 cm deep for tests with one or two beetles, and 14 cm wide x 1.9 cm deep for tests with five beetles. A disc of white filter paper covered the bottom of each dish to add traction for the beetles. A 3 ml glass vial, filled with distilled water and stoppered with cotton was placed in each dish to provide free moisture. To distinguish individual beetles we painted a dot on their dorsum with white paper correction fluid . The first tests were initiated on July 22 and continued for three months. Five times daily, every two hours, dishes were checked and the position and activity of each beetle noted. After two months, only daily observations were made. Some additional tests were started later and these were continued for two weeks to two months . Shelter Selection . Both black and white paper shelters were provided to find out which the beetles preferred. These were made by rolling a 8 cm x 6 cm piece of construction paper into a tube starting at the shorter side and then bending one end back about 1 cm and making a tight crease (Fig. 2). Different species of scale insects belonging to two families and four genera were tested for shelters also, so a choice of a natural shelter (an old female scale exoskeleton) or an artificial shelter (paper) was possible. Because of the time of year, only empty old female exoskeletons of P. hemic ryphus were available. Fresh specimens, both prereproductive and postreproductive of Kermes kingi Cockerell ( Kermesidae ) , an oak infesting gall-like scale insect; Toumeyella liriodendri (Gmelin) or tulip tree scale (Coccidae) ; and Ceroplastes ceriferus (Fabricius) or Indian wax scale (Coccidae), still attached to 3 or 4 cm long twigs, were placed in various dishes. The twigs on which these were found were kept fresh by immersing their lower part in 3 ml glass vials filled with distilled water and stoppered with cotton. BEHAVIOR OF ANTHRIBUS NEBULOSUS 239 Fig. 1 A. nebulosus on Norway spruce, Picea abies twig, a common resting and grooming site. Fig. 2 Example of petri dish used to observed, nebulosus in the laboratory with black and white paper shelters, spruce twig in water vial, oak twigs, Quercus rubra , with Kermes kingi scale insects and empty Physokermes hemicryphus exoskeleton lying on filter paper. 240 THE VIRGINIA JOURNAL OF SCIENCE Food Consumption . Food made available to the beetles included scale insect adults, nymphs, and eggs; honeydew produced by scales; artificial honeydew; sugar-water and some sooty mold fungi on twigs and leaves. Kermes kingi, T. liriodendri and C. ceriferus produced eggs and nymphs while in the dishes with the beetles. An infestation of Planococcus ci tri (Risso), the citrus mealybug, on Begonia leaves provided all stages of this scale insect also. Because P. hemic ryphus adults were dead, nymphs were collected nearby and introduced attached to spruce needles. Honeydew was produced by the Kermes , Toumeyella and Planococcus . Two artificial honeydew solutions, one with a concentration of .025% by weight and the other at .05% were prepared using the following amino acids: lysine, glutamic acid, aspartic acid, alanine, valine, arginine, glycine, isoleucine, proline, threonine and tyrosine. To each solution a mold inhibitor (supplied by Carolina Biological Supply Co. for Drosophila medium) was added at .025% concentration. Sugar (sucrose) was added using 5% and 10% concentrations for the two solutions. A plain 10% sucrose solution was also made available to the beetles in one dish. Sooty mold fungi were present in two tests. The first was a very dense natural fungal growth on a 6 cm section of a tulip tree twig ( Liriodendron tulipif era L. ), which had a lot of dried- up honeydew on it from the T. liriodendri scale insects infesting it. The second was a flat encrustation of sooty mold on oak and Camellia leaves. Identification of the fungi showed Cladosporium sp. to be present in all samples. Other fungi found associated were Alternaria sp., Aureobasidium sp., Bispora sp., Diplococcium sp . , Fusicladium sp . , Helminthosporium sp . , and Spilocaea sp . Twigs of Norway spruce, red oak ( Quercus rubra L.), and tulip tree, which could be present in the beetles' natural environment, were introduced into the dishes with the beetles to see how they would react to them. Spruce twigs had no scale insects on them. All oak twigs had leaves removed and had one or more Kermes adult females attached. The tulip tree twig was covered with sooty mold fungi and had Toumeyella scales on it as described above. Weight, Size and Sex Determination . To find the average weight of the beetles, 74 were weighed all together on a Mettler balance (accuracy to 0.1 mg.). Then the 10 specimens appearing largest and the 10 smallest were weighed individually to find the range. Length and width were measured with an Olympus binocular dissecting microscope and a slide calibrated to 0.1 mm. After measurements, they were killed in Weaver's Formol Fixative: 40% formaldehyde, 5 ml; glacial acetic acid, 2.5 ml; chloral hydrate, 20 g; distilled water to 100 ml. Dissections were made to determine their sex. Spring Observations Eighteen beetles were removed from their outdoor overwintering cage on April 3, 1982, and brought indoors for observation. They were kept in the refrigerator at 3°C until April 11 and placed in four 14 x 1.9 cm plastic petri dishes each containing 4 or 5 beetles. A dot of "Liquid Paper" was painted on their dorsum so each could be identified individually. When 2 beetles died and 4 had escaped, the remaining 12 insects were placed in new dishes. Four beetles, known to be mated females BEHAVIOR OF ANTHRIBUS NEBULOSUS 241 from observing copulations, were transferred to separate plastic petri dishes to observe egg laying habits. The other eight beetles were divided equally between two dishes. Food was supplied to the beetles as in the fall study. Scale insects used in the spring were: Parthenolecanium quercif ex (Fitch) on Quercus phel los L. twigs, Physokermes hemicryphus Dalman on Picea abies (L.) Karst. twigs, Coccus hesperidum L. on cyclamen leaves (all Coccidae); Kermes kmgi Cockerell (Kermesidae) on Q- rubra L. twigs. Honeydew produced by these scale insects and sooty mold fungi (containing Cladosporium sp.), growing on the twigs and leaves were available as food. The same type of water vial used before was provided also. No paper shelters were put in the dishes. Observations were made 4 times daily using a dissecting microscope. Hygrothermograph records showed the temperature in the laboratory varied from 17° to 33°C (X = 19.5°C), and the relative humidity from 30 to 90% (X = 54%) . RESULTS AND DISCUSSION Table 1 summarizes the fall activities of 18 Anthribus nebulosus adults from eight petri dishes. Since no difference in activities was found when more than one beetle was put in each dish, results are expressed for all the beetles collectively. The last category in the table, labelled "Not Visible", includes three separate conditions. Sometimes even a thorough search did not reveal the beetle. Also, if the beetle was inside a paper shelter, it could not be seen unless the paper was unrolled, which meant disturbing the beetle, therefore shelters were not opened at every observation. The third circumstance was when a beetle escaped. General Activity . Most beetles spent about 6% of their time moving around the dish, sometimes walking very rapidly, approximately the same amount of time they spent resting (Table 1). This latter category included only observations of beetles resting in the open. When they were holding still on a twig or on the water vial, etc., this was recorded elsewhere. Beetles were found on their backs kicking about 4% of the time. This usually was the result of their falling off the lid and landing on the bottom of the dish where the smooth surface of the glass made it difficult for them to turn over. Covering the bottom of the petri dishes with filter paper helped alleviate this problem. When beetles were found lying still on their sides or backs, they were usually moribund. The average for this category was about 7% of the observations (Table 1), although there was a wide range among the beetles. Shelter Selection . Beetles showed a wide range of responses to the shelters provided, from some never entering a shelter to one going in August 5 and staying until mid-November when the experiments were terminated. The latter probably continued summer diapause into winter diapause, but most beetles moved in and out of the shelters rather than staying inside. A definite preference was noted for black paper shelters over white ones (Table 1), so only black shelters were used in later experiments. Beetles moved in and out of the black shelters more often in late summer and tended to spend more time 242 THE VIRGINIA JOURNAL OF SCIENCE Table 1. Activities of Anthribus nebulosus beetles in the fall as % of observations for each activity (N = 18 beetles). Type of Activity Mean Range In the open space: moving around dish 5 . 6 3.8 - 9.5 resting on legs 5 . 7 0 - 11.3 on back, kicking 3 . 7 0.4 - 17 . 0 lying on side or back 7 . 5 0 - 41 . 5 Inside shelter: white 0.6 0 - 1 . 8 black 21 . 4 0 - 47 Drinking : water 0.9 0 - 3 . 8 artificial honeydew 3 . 5 0 - 9 . 5 (5% sugar) By Physokermes exoskeleton : on top 0.08 0 - 1 . 0 inside (for shelter) 6.3 0 - 79 chewing on 0.02 0 - 0.4 On spruce: on twig 12 . 7 6.6 - 25 on wet cotton stopper 0.5 0 - 4 On oak: on twig 19 . 8 0 - 30.6 on wet cotton stopper 0.72 0 - 2 . 5 feeding on Kermes 1.5 0 - 4.4 moving Kermes egg 0. 12 0 - 1 . 6 at or on Kermes 8. 1 0 - 18.2 partly inside Kermes 0.54 0 - 4.3 entirely inside (for shelter) 3.3 0 - 39.9 at honeydew drop 1.2 0 - 9. 1 chewing sooty mold 0.06 0 - 1. 1 Not visible: (probably in shelter) 7.2 0 20.5 BEHAVIOR OF ANTHRIBUS NEBULOSUS 243 inside in the fall. Time inside a shelter ranged from 2 hours to 1-1/2 months. The beetles did not use the Physokermes exoskeletons for shelters as much as the black paper ones (Table 1); only five beetles used these at one time or another. Time inside Physokermes exoskeletons ranged from two hours to a few days, except for one beetle which used an old exoskeleton for a permanent shelter. Old Toumeyella females were made available to beetles in a separate test along with Physokermes and Kermes exoskeletons. Of five beetles, two chewed holes in the exoskeleton of a postreproductive Toumeyella female and one eventually went inside and stayed 18 days (until the experiment was terminated) . As with the other shelters, the beetles showed no distinct patterns in the use of the post- reproductive Kermes females. Eight beetles went inside for varying lengths of time, and three remained inside, with two beetles sharing the same shelter in one case for 21 days. Length of time spent inside Kermes exoskeletons ranged from two hours to one month. Apparently the wax present on the live Ceroplates females kept the beetles from chewing on this scale, as none were seen attempting to make a hole in any of these scales. This is an important finding that might prevent successful utilization of the beetles against wax scales in the Eastern United States. Eight of the 31 beetles eventually sought permanent shelter, i.e. they went in and were not seen outside again. The time of entry ranged from August 5 to October 28. Five of these chose a female scale body for their permanent shelter and three chose a black paper shelter. Of the three latter, however, two had no other choice because only Ceroplastes scales were present and these were covered with wax. In one case a beetle chose a Kermes which was already occupied by another beetle. In four of the five cases where female scale bodies were used for shelter, beetles had to chew a hole in the female's exoskeleton to enter it. Jablonowski (1916) reported that the adult beetles stayed under the exoskeletons of Parthenolecanium corni (Bouche) for overwintering in Hungary. Food Consumption in the Fall . Naturally produced honeydew was consumed by the beetles no matter which species of scale insect was the source, although not every beetle was observed feeding on it (Table 2). Beetles fed on both fresh and dried-up honeydew. Artificial honeydew was also ingested and the solution with the higher sugar content (10%) was preferred (Table 2). When a 10% sucrose-distilled water solution was introduced into a dish simultaneously with the 10% sucrose-amino acid solution, beetles showed no preference (Table 2). The beetles in this dish were never actually seen drinking natural honeydew, although they did spend a lot of time on the Begonia leaf which had honeydew from the mealybugs (P. ci tri ) . We are assuming that they were attracted to the Begoni a leaf, not only as a shelter, but also as a source of honeydew. Although first instars of Kermes , Toumeyella, and Physokermes (dead and alive) were present with the beetles for about two weeks, no interest was shown in them by the beetles. Even though first instars crawled over the beetles, including their antennae, the beetles did not respond. No feeding on live scale insect adults was seen either. Beetles did chew on old dry exoskeletons of Physokermes and Kermes , and on shriveled up 244 THE VIRGINIA JOURNAL OF SCIENCE Table 2. Food consumption of Anthribus nebulosus in the fall. Mean % of Range Type of Food Observations (%) Natural honeydew: Toumeyella liriodendri 0.38 0 - 1 . 9 Kermes kingi 0.82 0 - 9 . 1 Planococcus citri* Artificial honeydew (with amino acids): 8.9 0 - 27 . 4 5% sugar 2.9 0 - 9.5 10% sugar 4.8 0 - 10.7 Sugar water (10%) 4.8 0 - 10.7 Sooty mold fungi: on oak 0.04 0 - 1.1 on tulip tree Dry exoskeletons of postreproductive female scale insects: 5.3 0 - 15 . 8 P. hemicryphus 0.02 0 - 0.4 K. kingi 1.3 0 - 4.4 T. liriodendri 2.3 0 - 7 . 7 C. ceriferus (without wax cover) 4.4 0 - 9. 1 *Not always actually eating the honeydew during observation. Ceroplastes and Toumeyella exoskeletons which had a lot of dried up honeydew on them (Table 2). Eggs were moved by beetles when making a shelter, but were not preyed upon. Sooty mold fungi (including Cladosporium sp.) present on the tulip tree twigs were eaten by the beetles (Table 2). The fungus-covered twig appeared to be a favorable habitat, not only for feeding but for resting and grooming, as the beetles spent a good portion of their time on it. However, the fungal growth on Camellia and oak leaves was not fed on. Beetles appeared to feed on fungi when a good growth of mycelia was present, as on the terminal bud of an oak twig. Food Consumption in the Spring . In contrast to our fall observations, both male and female adult beetles ate all stages of scale insects in the spring, including a male pupal case of P. quercif ex . Teneral females of P. quercif ex were most often their food source (Table 3). The beetles readily chewed a hole in either the anterior or posterior end and consumed the entire female in one day, with two beetles sometimes feeding simultaneously on one female scale insect. Teneral females of P. hemic ryphus were also consumed, but were not chosen as often as P. quercifex . We did not see beetles eating teneral females of K. kingi , although it is likely that they do. BEHAVIOR OF ANTHRIBUS NEBULOSUS 245 Eggs of P. querci f ex were also eaten more often than those of K. kingi or P. hemic ryphus (Table 3). Beetles chewed a hole in the scale exoskeleton and then began eating the eggs, moving farther in as the eggs were consumed, rather than pulling the eggs out to feed on them. During the fall beetles were observed I pulling eggs out, not to eat but to use the emptied scale exoskeleton as a shelter. We recorded only a few instances of beetles eating nymphs of scale insects, of P. querci f ex and P. hemic ryphus , but not of K. kingi. The latter are probably eaten, but our observations were not extensive enough to detect it. Beetles also ate exoskeletons of postreproductive female scale insects. Beetles visited the honeydew produced by the C. hesperidum females on the cyclamen leaf much more often than the honeydew from the other scale insects. We are not sure why this source was preferred, but it would be interesting to find out if it had a higher sugar content than the other honeydews, because tests with artificial honeydew solutions in the fall showed that beetles chose those with a higher sugar content over those with a lower . Natural growths of sooty mold fungi on twigs were eaten by beetles, but not to the extent that scale insects were (Table 3). For the first time we observed beetles apparently consuming the plant exudate from a cut oak twig. This has never been reported before to our knowledge and should be investigated further. The larvae of these beetles are known in Europe and Eastern Kazakhastan (USSR) to feed on the eggs of at least 14 species of soft scales (Coccidae) - 5 species of Eulecanium, 1 Nemolecanium, 2 Parthenolecanium , 3 Physokermes , 1 Pulvinaria , and 2 Rhodococcus . They also attack 1 species of Kermes , family Kermesidae (Prell 1925; Schmutterer 1952; Matesova 1966; Klausnitzer & Forster 1976; Kosztarab & Kozar 1978). Telenga (1954) and Forster (1973) also listed first instar soft scales as their food source. Only scanty and conflicting literature records are available on the spring feeding habits of the adults before oviposition. For example, as food for the adult beetles honeydew was given by Schmutterer (1952) and Forster (1973), and remains of old hardened exoskeletons ("mummies") of adult females listed by Schmutterer (1952), Forster (1973) and Goanca et al (1974). In addition, Schmutterer (1952) and Forster (1973) named the leftover scale eggs, while Goanca et al (1974) listed overwintered scale insect nymphs. In contrast. Kiss (as reported in Illes et al, 1896) stated that the adults fed neither on scale eggs, nor on young nymphs. In Moldavia, USSR, (Goanca et al 1974), adult beetles fed until oviposition on overwintered nymphs of the scale insects Parthenolecanium corni (Bouche) and Eulecanium ti liae (L. ) (Schrank). After oviposition, the exhausted adult beetles frequently consumed honeydew, chewed into the scale insects ( Physokermes piceae Schrank) and fed on their eggs and on the leftover exoskeleton of the females in Germany (Schmutterer 1952). Our observations indicate that food preferences of adult beetles are different after overwintering, the primary spring food being live scale insects and their honeydew, with fungi constituting a minor part of their diet. Nevertheless, these As Per Cent of Total Number of Feeding Observations 246 THE VIRGINIA JOURNAL OF SCIENCE Cl c s_ a to a) * BEHAVIOR OF ANTHRIBUS NEBULOSUS 247 beetles have kept their original fungal feeding habit which is characteristic of the family Anthribidae or Fungus Weevils. The familyy primarily of tropical and semi-tropical distribution/ is common in tropical rain forests because of the high humidity which favors fungal growth. It is speculated here that A. nebulosus has adapted more recently to the temperate zones of Eurasia, where in the semiarid areas their primary food source of fungi is scarce. Thus, they may have secondarily adapted to feeding on scale insects to supplement their original diet. The same reason could be used to explain how some species of Anthribidae have adapted to feed on dried fruits in Australia and New Zealand. The slow-moving nymphs and their sedentary females in soft scales and Kermes appear to be ideal prey for this slow- moving and non-aggressive predator. Site Preferences . Although the surface area of the twigs present with the beetles in the fall tests was not measured, it was obvious that all of them combined did not make up an area equal to half of the filter paper area. Our observations indicated that beetles preferred being on twigs to being in the open (Table 1 ) • Interactions . Beetles were generally solitary and non-aggressive during the fall. When two met head on, they stopped and one or both would go around the other. Although four dishes (14 cm diameter) contained five beetles for six weeks, there was only one occasion when one beetle was seen interacting physically with another. In that case the first beetle had its front legs on the back of the second for a short while before moving away. When several beetles were inside the same black shelter, they seemed to be clustered, but this may have been because the shelter was relatively small. Close clustering was also observed by us when beetles were inside corrugated paper rolls during summer and winter diapause. Under natural conditions, 2 beetles occasionally use the same exoskeleton of a Physokermes female as a shelter. No signs of cannibalism were shown by adult beetles kept in captivity, or outdoors under natural conditions. Weight and Size . The average weight of a beetle in mid-November was 3.6 mg (N = 74). The average weight of the ten largest beetles was 5.9 ± 1.1* mg; of the 10 smallest, 2.1 ± 0.7* mg. The mean length of the ten largest beetles was 4.6 ± 0.1 mm; mean width 2.0 ± 0.1 mm. For the 10 smallest beetles mean length was 3.4 ± 0.2 mm, and the mean width 1.4 ± 0.1 mm. Secondary Sexual Dimorphi sm . The ability to distinguish the sexes by external features would be very helpful, and two characteristics have been suggested. Dr. A'rpad Szentesi (Hungarian Research Institute of Plant Protection) used the presence of a slight curvature at the tip of the abdomen in males (personal communication). A second character used by Dr. Barry D. Valentine (Ohio State University) is sternites IV and V being longer along the midventral line in females (personal communications). We tried sexing the 10 largest and 10 smallest beetles by using both of these methods, then dissected them to determine sex. Overall size turned out to be unacceptable for determining sex in our sample since 50% of both large and small beetles were males. Forster (1973) in Germany also found no differences in size between the sexes. The curvature at the tip of the abdomen 248 THE VIRGINIA JOURNAL OF SCIENCE was unclear in 25% of the beetles in our sample. However, when the abdomen was definitely turned down, in every case the beetle was a male. One beetle classified in the "abdomen not turned down" category was found to be a male also. Thus this character is not very reliable for beetles which have no curvature or only a very slight curvature of the abdominal tip. A larger sample of beetles should be studied to test this correlation. Measurements of the widths along the midventral line of abdominal sternites IV and V showed females averaging 0.42 ± 0.11 mm for IV and V combined, and males 0.32 ± 0.06 mm. A student's t-test on these data revealed no significant difference at <*=.05. We also found that when sternite IV was wide, sternite V was not correspondingly wide. Thus, from our limited sample we concluded this method of determining sex was not reliable either. Grooming . The tarsi, antennae, palps and other mouthparts became sticky when feeding on honeydew, and also after holding mouthparts for some length of time just over the anal plate area of soft scales. Only self-grooming was observed, never mutual grooming . After feeding on honeydew produced by P. quercifex, the beetles were observed cleaning the tarsi of the anterior legs between their mandibles by pulling the tarsi repeatedly through the mouthparts. The same procedure continued next on the midleg tarsi. The insects were able to reach and clean even the femora of the midlegs between their mandibles by lowering and bending the head underneath the body. Antennae were cleaned after feeding on honeydew by bending these in a semicircular fashion to fit between the mandibles and pulling them repeatedly between the mandibles with a combing motion. Mating Behavior . Nine beetles of the original 18 present in the spring tests were observed in copula, and twenty-six copulations were recorded, the first on April 14 and the last on May 3. No time of the day was preferred. Of the four male beetles, only one mated with more than one female, the number of copulations per male beetle ranged from one to six. All five females had just one partner except for one which mated with two males. Number of copulations per female was one to four. No beetle copulated more than once in the same day during the hours of observation. Length in copula ranged from 3 minutes to 35 minutes. The male mounted the female from the back, clasping her with his first and second pairs of legs, and rapidly tapping her back with his antennae. The female would sometimes begin walking around the dish during mating, dragging the male along. No difference in size was noticed between members of copulating pairs. Development . Schmutterer (1952) reported that the female beetle chewed a hole in the still semi-soft exoskeleton of the female scale insect, and laid an egg inside the brood chamber of the scale and closed the wound with a secretion. The egg laying sites showed later only as darker spots on the female's exoskeleton. Other authors did not elaborate on the mode of egg laying . Only two of our five mated females laid an egg that could be visually detected, one on May 7 and one on May 11. The preoviposi tion period after mating was 8 days in one case and * standard error. BEHAVIOR OF ANTHRIBUS NEBULOSUS 249 about 15 days in the other. One egg was laid on top of a dead P. quercif ex teneral female. The egg was oval, 0.8 mm long and 0.4 mm wide. Its color was light yellow (Munsell ,1965), hue 7.5, value 8, chroma 4. This egg was moved by us from the open to under a large P. quercif ex female with many eggs under her, but it never hatched. A second egg was laid on a spruce twig, a short distance from a P. hemi chryphus female. This egg remained the same for three days and then changes were noted. The outlines of the body and red eye spots appeared. By the fourth day the egg hatched and a clear case was left on the twig. The larva chewed a hole in the exoskeleton of a Physokermes female and went inside to feed on the eggs. After just one day the beetle larva disappeared and we were unable to find it again. A second larva, that hatched from an undetected egg, was found on May 7 under a P. quercifex female filled with eggs and with a large hole chewed on her lateral side. The length of the larval and pupal stages could not be determined separately for this individual beetle because it was not always found when observations were made. However, because the pupa was discovered on May 30 and the adult emerged on May 31, the combined length of the larval and pupal stages were calculated to be about four weeks. It took about 6 hours for the newly molted adult beetle to change to its natural dark color. About 24 hours after emergence, it was eating a P. quercifex female, and the next day was feeding under a scale exoskeleton full of egg shells and nymphs of P. quercifex . On June 3 the beetle became motionless inside a P. quercifex exoskeleton which was detached from the twig and lying on the bottom of the dish. It remained there until June 18 when it was found lying still in the open. The larvae pupated in mid-June and adults emerged the end of June in Moldavia (Goanca et al 1974). Adults emerged from mid- July to early August in Eastern Kazakhstan (Matesova, 1966). Schmutterer (1952) and Forster (1973) in Germany found pupae in June- July and the adults emerged a few weeks later. Longevity . Of 12 adult beetles the average longevity after overwintering was 47 days (range 14 to 120 days) and was about the same for both males and females. ACKNOWLEDGEMENTS The authors wish to express their sincere appreciation to two professors in our Department, Dr. Donald G. Cochran, for the preparation of the artificial honeydew used in the experiment, and Dr. L. T. Kok, for critically reviewing the first draft of this manuscript; to Dr. R. Jay Stipes, professor of Plant Pathology and Physiology, VPI&SU, for identifying the sooty molds; also to Dr. Ferenc Kozar, Head, Zoology Department, Research Institute of Plant Protection in Budapest, for the assistance with the collecting of the Anthribus beetles in Hungary; and to Dr. Peter B. Schultz, Entomologist of the Virginia Truck and Ornamental Research Station, Virginia Beach, Virginia, for supplying live scale insects as food for our beetle colonies. Names of other scientists who also assisted this study are given in the text. This was part of our state funded research, supported by VPI & SU (Project 2024390) and part of a National Science Foundation supported research (DEB-76-09676). 250 THE VIRGINIA JOURNAL OF SCIENCE REFERENCES CITED Forster, ' G. 1973. Zur Biologie und Morphologie von Anthribus nebulosus Forster (Col., Anthr.). Entomol. Nachr. 17: 117-121 . Goanca, I. K. , E. S. Sugon j aev and E. M. Danzig. 1974. Scale insects and soft scales of Moldavia and their natural enemies (In Russ.). Izd. Kartya Moldov. , Kisinev, 112 p. Ill'es, N. , J. Havas, G. Horvath and J. Vadas. 1896. The Black Locust Scale problem in Hungary (In Hung.). National Forestry Assoc., Budapest. 207 p. Jablonowski, J. 1916. The scale insects of grapes and other economic plants 1-120. (In Hung. ) . Ki serletugyi Kozl . 19(2): Klausnitzer, B. and G. Forster . 1976. Zur Kenntnis der Parasitierung weiblicher "Brutblasen" der Fichtenquirlschildlause Physokermes piceae Schrk. und Physokermes hemicryphus Dalm. Hercynia N. F., Leipzig 13(1): 85-92. Kosztarab, M. and F. Kozar. 1978. Scale insects--Coccoidea (In Hung.). Fauna Hungariae No. 131, Vol. 17(22): 1-192. Kosztarab, M. and F. Kozar. 1983. Introduction of Anthribus nebulosus (Coleoptera: Anthribidae ) , in Virginia for control of scale insects: A Review. Va. J. Sci. 34(4). Matesova, G. Ya. 1966. Beetles of the genus Brachytarsus (Coleoptera, Anthribidae) that are enemies of soft scales (Homoptera, Coccoidea) in Eastern Kazakhstan. Entom. Rev. 45(2): 141-142. Munsell Book of Color. 1965. Pocket Ed. Munsell Color Co., Inc. Baltimore, Md. 41 color charts. Prell, H. 1925. Beitrage zur Biologie des grauen Schi ldlausrusslers ( Anthribus nebulosus Forst.). Z. Forst- u. Jagdwesen 57: 245-250. Schmutterer, H. 1952. Die Okologie der Cocciden (Homoptera, Coccoidea) Frankens (Pt. 3). Ztschr. f. angew. Entomol. 33: 65-100. Telenga, N. A. 1954. Parasites and predators of plum and white locust tree scales (In Russ.). In Biological Control of Pest Insects. Akad. Nauk Ukr. SSR. 5: 110-135. Virginia Journal of Science Volume 34, Number 4 Winter, 1983 / V Annual Production and Phytoplankton Studies Within the Barrier Islands of Virginia, U.S.A. Kneeland K.^Jesius, Harold G. Marshall, and Charles K. Rutledge Department of Biological Sciences Old Dominion University Norfolk, Virginia 23508 ABSTRACT Phytoplankton primary production rates were determined within the Barrier Islands complex of Virginia. The total production rates followed seasonal changes in water temperature. The production rates ranged from a low of approximately 1 mg C/m^/hr in February to a high of 290 mg C/m2/hr in June. Total annual production rate was 178 g C/m2/yr. The concentration of phyto¬ plankton varied seasonally, but did not coincide directly to the periods of peak production. INTRODUCTION Previous investigators have indicated the importance of phytoplankton to productivity within estuaries along the eastern coast of the United States (St.ross and Stottlemeyer , 1965; Thayer, 1971; Thomas, 1966) . The signifi¬ cance of detrital substances produced by phytoplankton within Georgia estuaries has been further discussed by Haines (1977) . She indicates the total input of organic matter to Georgia estuaries by the phytoplankton is greater than what has been generally considered, and places a lesser emphasis on the contributions of organic matter by the Spavtina salt marsh to these estuaries. Productivity rates in estuaries along the United States east coast have generally been higher than those taken from stations over the shelf (Riley, 1956; Smayda, 1957; Sellner et al. , 1976) . One of the highest values was recorded by Thomas (1966) when he reported 546 g C/m2/yr in the Altamaha River in Georgia. Considerably lower rates have been reported in the Beaufort, N.C. estuary where annual pro¬ duction rates of 53 and 67 g C/m2 were determined by Williams (1966) and Thayer (1971) . In a South Carolina estuary, Sellner et al. (1976) found productivity rates generally followed the annual temperature cycle, having a range between 6.4 to 234 mg C/m2, with the total annual production of phytoplankton calculated to be 346 g C/m2. One significant estuarine area of the east coast that lacks productivity information is the Barrier Island complex of Virginia. Beginning at the Chesa¬ peake Bay entrance and extending 120 km northward to the Maryland bor*der, the barrier island chain encloses approximately 700 km2 of low land salt marsh that is interlaced with numerous channels. To determine the primary produc¬ tion rates within the channels of this area, a one year study was undertaken in conjunction with a total phytoplankton survey. Comparisons were made between these primary production rates, and the composition and concentration values of the phytoplankton. 252 THE VIRGINIA JOURNAL OF SCIENCE METHODS The study area is located near the town of Oyster, Virginia, approxi¬ mately 23 km north of Cape Charles. Eight stations were established in waterways that included Sand Shoal,' Eckichy, and Mockhorn channels located between Oyster and Cobb Island (Fig. 1) . Samples were taken at mid-channel locations during 10 collection trips spaced between November 1978 and October 1979. A 500 ml water sample was taken at the surface at each station for phyto¬ plankton analysis. The samples were preserved immediately with buffered forma¬ lin and returned to the laboratory for settling. A modified Utermohl method was followed in which the samples were allowed to settle, then siphoned to a 20 ml concentrate. The concentrate was transferred to a settling chamber and subsequently examined with a Zeiss inverted plankton microscope. The entire bottom slide was scanned, with the exception of the more highly concentrated samples where subsamples were used. Phytoplankters were identified and counted to obtain cell numbers per liter. Twenty-five ml water samples were collected from the surface for primary production analysis. These samples were placed into 125 ml glass-stoppered bottles and . inoculated with 3 to 5 yCi NaHl^co^ (specific activity 56 yCi/y mole) and incubated from 3 to 5 hr in the time span 0900 to 1500 hr in natural light. Duplicate light and dark samples were set up. These samples were fil¬ tered through 0.45 ym millipore filters at vacuum pressure less than 60 mm of Hg. Filters were washed with 0.1 M HC1 and placed in scintillation vials con¬ taining 10 ml of scintillation fluid (6 g of 2 , 5-diphenyloxazol and 100 g of naphthalane/liter 1,4-dioxane) . One ml aliquots of acidified, air-bubbled filtrate were pipetted into scintillation vials containing scintillation fluid. The cT4 found in these vials was used to determine the dissolved organic C44 (DOC) released. Annual production rates were calculated by multiplying the mean f ixation/m2hr • 12hr/d* 365d/y . Salinity and temperature values were obtained with a portable Beckman salinometer (Model R55-3) . A model 126A Photovolt portable pH meter was used for pH readings, and a standard sized secchi disc for visibility. RESULTS Total production rates followed seasonal changes in water temperature (Fig. 2) . The samples showed a decrease in the total productivity rates from the Fall of 1978 into Winter, which corresponded to a decrease in water temperature. This productivity was from 9.5 mg C/m2/hr on 15 November 1978 to 1.2 mg c2/m2/hr. on 12 February 1979. During Spring the total rate of productivity increased from 16.8 mg C/m^ in April to 290 mg C/m2/hr in June 1979, which corresponded to an increase in water temperature. From 13 August to 3 October 1979 the total productivity rates declined. Maximum rates were recorded on 24 June 1979 with Station 7 showing the highest rate, 348.6 mg C/m2/hr (Table 1) . Minimum rates were observed at the times of collection on 21 February 1979 and 19 March 1979 for all stations sampled. Monthly differences occurred in the amount of C44 fixed by the phytoplankton at the various stations, with the annual production rate for this study 178 g C/m^/yr. The concentration of phytoplankton varied seasonally, having growth pulses occurring in January, March, and August. Maximum numbers were recorded on 19 March 1979 with average counts for all stations approximately 1.89x10^ cells/1 (Fig. 2) . The seasonal lows occurred in December, February, and June. Species composition was predominated by forms characteristic of shelf populations and previously reported by Marshall (1976) , and Marshall, Nesius and Cibik (1981) . These mainly were nanoplankton sized diatoms that included LeptocyHndrus danicus L. minimus 3 Asterionella giaciaiis3 Skeletonema oostatum 3 Rhizosolenia delicatula and Cylindvotheca clostevium. Diatoms were dominant during all seasons. The winter peak of January consisted mainly of nanoplankters dominated by PHYTOPLANKTON PRODUCTION WITHIN VIRGINIA BARRIER ISLANDS 253 Table 1. Total production rate for each station (mg C/m /hr) . THE VIRGINIA JOURNAL OF SCIENCE 00 1 — 1 CTV rH CO CM CO CTv CO vO 00 CO rH oo 00 rH CO 00 • • • 1 • • • • • . rH m CM i — 1 CTv o CM 00 CO CM CM m rH CM co CM vO CTv CM iH 00 CM rH VO rH 00 CO rH CM CO • • • 1 • * • • • • CM m CM rH rH rH CTv rH CM Cs VO CM CO O CO i — 1 CO 00 CM in rH m 00 'cr rH 00 VO • • • 1 • « • 1 • 1 CO 00 rH co CTv rH rH rH iH co rH CO rH VO rH CO CM CM 00 co vO C- • • • 1 • • « • • 1 CTv CO rH CM co o 00 vO I— 1 rH iH 'd- CM CO o i— 1 •. Cn W o C u c 00 4-1 < o 0) CO 0> eg q. eg 3 CJ Q 2; Q ■-> F^i S < S •o < o m CM o rH CTv CO CM CO co i— i rH rH CM rH CM CM CM I— 1 PHYTOPLANKTON PRODUCTION WITHIN VIRGINIA BARRIER ISLANDS 255 Leytocylindrus dccnicus , L. minimum , Asierionella glaoialis , and Rhizosolenia delicatula, Basically the same populations, but in much lower concentrations, were noted in the February collection which coincided with low productivity values and temperatures of 0°C. March was accompanied by rising temperatures and a major increase in phytoplankton numbers, but no major increase in produc¬ tivity was noted at that time. The phytoplankton composition in March consisted of a combination of the same species as the winter peak, plus several Chioretta spp. , and other ultraplankton components. The prominent diatoms consisted of filamentous or colonial forms. Clusters, or groups of other phytoplankters were typically found, indicating conditions favorable for rapid multiplication. Subsequent collections from April through June indicated a decrease in cellular counts and a transition to a larger sized diatom community. Many of the past prominent species were still among the dominants ( S . costatum , R. delicatula , L. minimus, R. setigera) , however several Chaetoceros species and Guinardia flaccida were also common. This seasonal successional pattern was similar to that outlined by Margalef (1958) with the exception that there was no significant increase in the concentrations of dino- phyceans into summer. Instead the summer dominants were mainly larger sized diatoms (e.g. Chaetoceros spp. Corethron hystrix, Lithodesmium undulatum) , with the appearance of the cyanophycean Nostoc commune, which persisted into fall. The highest productivity rate occurred in June. This coincided with a decreasing population level, which averaged 5x10^ cells per liter at each station. This was followed by a moderate fall pulse where the same nanoplankton forms again became dominant. SUMMARY The monthly primary production rates followed a seasonal pattern that corresponded closely to temperature. The productivity rates decreased with a decline in temperature into winter and rose with the increased temperatures into summer, then dropped again into late fall. The phytoplankton levels were generally high throughout the study, exhibiting seasonal changes in popu¬ lation concentrations, having pulses in winter, spring, and fall. The occurrence of the periods of greater phytoplankton concentrations did not appear to effect the periods of peak productivity, which did not coincide with maximum population size. The channels in this study were dominated by a diato- maceous flora, characteristic of forms found over the shelf. The data indicates an annual production rate for these channels to be 178 g C/m^/yr. This rate is within the range found by other investigators for other coastal estuaries on the United States east coast (Smayda, 1957; Thayer, 1971; Williams, 1966; Thomas, 1966; and Sellner et al. , 1976). LITERATURE CITED Haines, E. B. 1977. The origins of detritus in Georgia salt marsh estuaries. Oikos 29: 254-260. Margalef, R. 1958. Temporal succession and spacial heterogeneity in natural phytoplankton. In: Perspectives in Marine Biology, University of Cali¬ fornia Press, Berkeley, California, pp. 323-349. Marshall, H. G. 1976. Phytoplankton distribution along the eastern coast of the USA. I. Phytoplankton Composition. Marine Biology 38: 81-89. Marshall, H. G. , K. K. Nesius, and S. J. Cibik. 1981. Phytoplankton studies within the Virginia Barrier Islands. III. Seasonal Study of Phytoplank¬ ton within the Barrier Island Channels. Castanea 46(2): 89-99. Riley, G. A. 1956. Oceanography of Long Island Sound, 1952-1954. IX. Pro¬ duction and utilization of organic matter. Bull. Bingham Oceanogr. Coll. 15: 324-344. 256 THE VIRGINIA JOURNAL OF SCIENCE Sellner, K. G. , R. G. Zingmark, and T. G. Miller. 1976. Interpretations of the cl ^ method of measuring the total annual production of phytoplankton in a South Carolina estuary. Botanica Marina 19: 119-125. Smayda, T. J. 1957. Phytoplankton studies in lower Narragansett Bay. Limnol. Oceanogr. 2: 342-359. Strickland, J. D. H. and T. R. Parsons. 1968. A Practical Manual of Seawater Analysis. Fish. Res. Bd. Canada: Bull. 125, Ottawa, Canada, p. 203. Stross, R. G. and J. R. Stottlemeyer . 1965. Primary production in the Patuxent River. Chesapeake Sci. 6(3): 125-140. Thayer, G. W. 1971. Phytoplankton production and the distribution of nutrients in a shallow unstratified estuarine system near Beaufort, North Carolina. Chesapeake Sci. 12: 240-253. Thomas, J. P. 1966. The influence of the Altamaha River on primary production beyond the mouth of the river. M.S. thesis. Department of Zoology, Univ. Georgia, Athens, Ga. Williams, R. B. 1966. Annual phytoplanktonic production in a system of shallow temperate estuaries. In: H. Barnes (ed.), Some Contemporary Studies in Marine Science, George Allen and Unwin Ltd., London, pp. 669-716. Virginia Journal of Science Volume 34, Number 4 Winter, 1983 / Seasonal Changes in the Phytoplankton of Lake Chesdin, Virginia with Ecological Observations B. R. Woodson, Jr.7, and KennethiSeaburg^ Lincoln University, Lincoln University, Pa. 19352 and Xavier University, New Orleans, La. 70125 Abstract The phytoplankton of Lake Chesdin was observed to be quite diverse with 339 species and varieties identified. The dominant alga, Melosira granulata var angustissima displayed large pulses during autumn and late spring at the shallower collection sites. The seasonal range in physical ecological factors such as sunlight, water temperature, and turbidity resulted in considerable variations in phytoplankton standing crop. The lowest standing crop of 45 cells/ml was observed in January, 1975, while the largest standing crop ( 3500 cells/ml) occurred during October, 1974. Water temperatures ranged from an average of 3QC during late summer to 3.2C during midwinter which resulted from the shallowness of the lake, the small drainage area of the Appomattox River above the lake, and the short hydraulic retention time of the lake. Introduction Lake Chesdin, Virginia, is a man-made reservoir created by the damming of the Appomattox River in 1967. Since the lake serves a vital role to the Tri-Cities of Colonial Heights, Hopewell, and Petersburg, Virginia, by being its principal source of public water supply, information regarding its relative ecological condition would be invaluable. One method of determining the ecological condition of any lake is to make an annual study of the phytoplankton, the primary producers in most aquatic ecosystems. Therefore, the purposes of this study were: 1. To identify all algal species encountered in the plankton of Lake Chesdin, 2. To monitor and quantify the populations of individual algal genera, and, 3. To relate populations of the algal genera encountered to seasonal ecological factors that might influence their populations . description of the Lake Lake Chesdin is located at approximately 77° 35' West longitude and 37° 15' North latitude or about 17 km west of Petersburg, Virginia. 258 THE VIRGINIA JOURNAL OF SCIENCE The lake (Fig. 1) follows the flood plain of the Appomattox River which gives the body of water a long (28km), narrow (less than 1 km) shape with a surface area of 12.95 km.^ The dam impounds about 90 X 106 m^ of water with the maximum depth of 14 m occurring at the dam. The upper 12 km of Lake Chesdin is a broad, extremely shallow area, often less than 2 m deep. Except during extremely dry summers, the water level exceeds the height of the dam which imparts a flowing nature to the impounded water and, therefore, a short hydraulic reten¬ tion time. The flowing nature of Lake Chesdin does not allow the development of a strong thermocline. However, a clinograde dissolved oxygen stratification does occur with the bottom becoming anaerobic. Materials and Methods Five collection sites (Fig. 1) were chosen with regard to the general morphometry of the lake. Qualitative phytoplankton samples were obtained by trolling at low speed in a motor-driven boat with a No. 25 mesh phytoplankton tow net over an unmeasured distance. The resulting concentrated samples were stored in 100 ml collection bottles and observed in a living condition. Species identifications of the diatoms were determined following acid clearing and mounting in Hyrax. Quantitative phytoplankton and water samples were collected by the use of a model 1200 modified Kemmerer sampler and transferred to collection bottles. Samples were collected at each site at the water's surface and depths of 1 and 3 meters. Phytoplankton counts were obtained by pipeting 50 ml sub-samples into centrifuge tubes and gently centrifuging at 1000 rpm for 20 minutes in an International Portable Refrigerated Centrifuge Model PR-2 that was calibrated to existing lake water temperatures. Algal residues were resuspended in a volume of 5 or 10 ml of filtered lake water depending upon the anticipated phytoplankton density. One tenth ml of this concentrated sample was placed in a Palmer-Maloney nannoplankton counting chamber and the algal genera identified, counted, and expressed as cells/ml for each of 3 replicate counts. The expression of algal counts as cells/ml rather than organisms/ml emphasizes the larger biomass of filamentous and colonial algae in comparison to unicellular forms. The small cell size of certain blue-green algae (e.g. Aphanizomenon) required that these be reported as filaments or colonies/ml. Water chemistry determinations (Table I) utilized the Hach water pollution test kit calibrated to standard methods (1). Dissolved oxygen and temperature were determined with a Yellow Springs International Oxygen meter Model 51-A. Hydrogen ion concentration was determined with a Beckman Zeromatic pH meter Model 6804. Results Appendix I lists the species of algae encountered as plankton in Lake Chesdin during the period from October 2, 1974, to August 4, 1975, with the months observed designated. Table II gives a breakdown of the identified algal species and varieties into important taxonomic groups. Many of the algal forms identified are not considered true plankton (euplankton) as for example many desmids (Zygnematales ) and diatoms (Bacillariophyceae ) , both of which were represented by species considered to be tychoplankton or meroplankton . The standing crop (Fig. 2) was dominated by the diatoms, in particular the centric diatom, Melosira granulata var . angustissima Muller. This filamentous species displayed significant pulses at sites 4 and 5 during both autumn and late spring when it often comprised 90% of the standing crop. Table I demonstrates the average number of cells observed during each "season" for the major taxonomic groups of phytoplankton in Lake PHYTOPLANKTON SEASONAL CHANGES, LAKE CHESDIN 259 Figure 1 Figure 2 260 THE VIRGINIA JOURNAL OF SCIENCE Chesdin. These seasons were based upon average water temperatures since this measurement best reflected the physical changes that occurred during the study period. The short autumn and spring seasons reflected the relatively small drainage area of the Appomattox River above Lake Chesdin as well as the shallowness of the lake itself. Table III lists the most abundant phytoplankton genera during each season. Physical and chemical analyses are given in Table I. The extremely high turbidity, often observed following period of heavy precipitation, indicates that the lake drainage area contains much disturbed land which is susceptible to erosion. During May, June, and July, 1975, the Appomattox River Water Authority treated the lake with large amounts of copper sulfate in order to inhibit the growth of potential "taste and odor" producing algae. These applications possibly accounted for the unusual fluctuations observed in the standing crop (Fig. 2) during these months. The use of copper sulfate as a potent algicide has existed for some time (2) and this compound has been shown to inhibit growth of Anabaena f los-aquae at lower concentrations than for the green alga Chlorella (3). The copper sulfate applications were devastating to certain algal forms (Melosira granulata and Asterionella formosa ) while other genera ( Scenedesmus and Ankistrodesmus ) were much more tolerant. The copper sulfate applications often masked natural variations in standing crop at the different collection sites (Table IV). Discussion The phytoplankton of Lake Chesdin closely resembled the description of North Carolina lakes and ponds by Whitford (14) although it differed markedly from the phytoplankton of the two natural lakes of Virginia, Lake Drummond (5) and Mountain Lake (6). Dystrophic Lake Drummond is dominated by the diatom, Asterionella formosa , which experiences large pulses during May, June, and September (5). Mountain Lake is dominated Table I Title: Average Seasonal Phytoplankton Composition and Physical-Chemical Results (Seasons based Collection Date. on Water remperature ) N = 15 Sampl es/ Fall Winter Spring Summer Collections 5 7 5 10 Inclusive Dates 10/2/74- 12/2/74- 4/16/75- 5/26/75- 11/26/74 3/26/75 5/19/75 8/4/75 Range of water temperature (°C) 9.6-20.0 4.4-8. 6 11.0-21.3 25.8-30.4 Total standing crop (cells/ml) 1456.1 184.0 1134.5 1342.4 Chlorophyta 20.9 4.3 87.8 181.9 Chrysophyta (w/o diatoms) 10.4 26.3 134.3 56.8 Bacillar iophyceae 1397.2 149.1 865.4 1045.5 Euglenophyta 21.9 1.7 18.5 43.6 Pyrrhophyta 0.3 0.1 2.7 6.9 Cyanophyta 5.4 0.0 1.3 2.4 Cryptophyta 0.0 2.5 24.5 5.3 Chemical-Physical Results pH 7.0 7.1 7.4 7.3 Average temperature ( C) 15.4 5.6 17.6 26.9 NO -H (mg/1) NHT-N (mg/1) 0.49 0.69 NA NA 0.38 0.89 0.81 0.41 PO^-P (mg/1) Calcium hardness (mg/e) .020 .029 .052 .027 15.3 12.9 16.1 19.8 Dissolved oxygen (mg/1) NA 12.1 8.7 7.4 Secchi disk (cm depth) NA 69.6 97.2 131.1 NA = Not Available PHYTOPLANKTON SEASONAL CHANGES, LAKE CHESDIN 261 Table II Title: Summary of Lake Chesdin Algae Encountered as Plankton Division Important Total Number Per cent Classifications of Forms of Total Chlorophyta Volvocales 13 152 44.88 Tetrasporales 3 Ulotrichales 2 Oedogoniales 2 Chlorococcales 72 Zygnematales 60 Euglenophyta 42 12.4 Chrysophy ta 116 34.2 Xanthophyceae Chrysophyceae Bacillariophyceae Pyrrhophyta 12 3.5 Cryptophyta 2 0.6 Cyanophy ta 15 4.4 339 99.9 Table III Title: Most Common Algal Genera by Season (cells/ml Average Values Based on 15 Samples/Collection Date Fall Winter Spring Summer Melosira 1388.4 Melosira 109.7 Melosira 732.7 Melosira 9 50. Eudorina 16.7 Synura 22.5 Synura 75.6 Synedra 63. Trachelomonas 13.7 Synedra 15.0 Synedra 74.8 Scenedesmus 33. Synura 9.3 Asterionella 14.2 Asterionella 32.3 Lagynion 28. Euglena 8.4 Fragilaria 5.7 Cryptomonas 24.5 Trachelomonas 19. Aphani zomenon 5. 3a Dinobryon 2.9 Uroglenopsis 22.3 Coelastrum 18. Synedra 4.1 Cryptomonas 2.5 Eudorina 16.6 Ankistrodesmusl8 . Dinobryon 1.4 Eudorina 1.2 Scenedesmus 14.4 Cyclotella 17. Pediastrum 1.2 Trachelomonas 1.0 Trachelomonas 13.9 Micratinium 13. Fragilaria 1.1 Mallamonas 0.9 Fragilaria 13.7 Dinobryon 11. Pandorina 13.6 Crucigenia 11. Synura 10. a = Filaments/ml Table IV Title: Total Standing Crop at Collect ion Sites by Season (cell/ml) Average Values Based on 3 Depths/Collection Date Collection Site Fall Winter Spring Summer Water Depth # 1 1537.5 586.0 1742.2 1208.4 12 m # 2 1692.4 600.1 2241.2 1430.6 9 m # 3 4623.2 817.7 3024.8 1794.6 5.5 m # 4 6065.8 333.4 3689.2 5208.4 3.7 m # 5 7839.4 427.9 6280.2 10505.4 2.2 m 262 THE VIRGINIA JOURNAL OF SCIENCE by the centric diatom, Cyclotella comta , and the green algae, Sphaero- cystis schroeteri and Planktosphaeria gelatinosa , but the phytoplankton densities of this oligotrophic lake are extremely low (6). Lake Chesdin is a soft-water lake (e.g. 15mg/l calcium hardness) and is very typical of the larger man-made lakes throughout the piedmont of the Southeastern United States (4). The dominant alga of Lake Chesdin, Melosira granulata var angustissima , has been reported (4) as being abundant in many rivers and lakes throughout this region. The high number of phytoplankton forms observed (339) indicates that the flora of the lake was quite diverse although many of the forms encountered were not euplanktonic . Since a member of the genus Melosira was the dominant plankter and the standing crop was relatively large, the phytoplankton association of Lake Chesdin fits Hutchinson's description (7) of a eutrophic diatom association. The seasonal variation in standing crop was extremely large, ranging from very low numbers of cells in midwinter to counts of 10,000 cells/ml during early autumn and late spring. The lowest standing crop (45.4 cells/ml) occurred when the average water temperature dropped to 3.2C on January 15, 1975, and undoubtedly resulted from the interactions of low temperature, low solar light intensity, and extreme turbidity. The largest standing crop occurred during October, 1974, a time when temperature (20-22C) and light intensity (moderate) were favotable for the growth of Melosira granulata var angustissima . The diversity of the autumn peak in standing crop was much lower than the late spring peak when significant populations of Scenedesmus , Coelastrum, Synedra , Dinobryon, Mai lomonas , Trachelomonas and Asterionella were observed with M. granulata which, in addition, was the host for large numbers of the minute epiphyte, Lagynion reductum . The fall pulse of M. granulata was accompanied by small populations of Eudor ina elegans , Euglena , Trachelomonas , and Synura . The chemical analyses of Lake Chesdin showed that the lake was rich in nitrate-nitrogen and orthophosphate-phosphorus at all times of the year which agrees with an earlier study (8). These observations suggested that nutrient limitation as a determining factor in seasonal variations of total standing crop was highly unlikely. The Appomattox River has been described (8) as being highly silicious which indicates that adequate amounts of silica were available for extensive diatom growth. The presence of these nutrients, along with the shallowness of the upper portions of the lake which allowed the photic zone to extend to the bottom, possibly resulted in the large populations of Melosira granulata var angustissima . In addition, the higher flow rate of the water in the upper portion of the lake could have aided the suspension of this species in the water column (7) and may help to explain the disparity in populations of this species observed at the different sites when conditions were optimal for its growth (Table IV). Although no large blooms of heterocystous blue-green algae were en¬ countered during the study period, qualitative net tows frequently revealed the presence of Anabaena spiroides and A. circinalis when surface waters approached 30C . During the autumn, Aphani zomenon f los-aquae was observed when temperatures were as low as 13C. Pearsall (10) has reported that lakes in England dominated by Melosira granulata usually experienced large blooms of blue-greens during late summer. Possibly the extensive treatments with copper sulfate during the spring and summer of 1975 inhibited the natural growth of the blue-green species as well as other forms, and it should be pointed out that no collections were made during late August and early September when water temperatures normally are their highest (Table V). The expansive, shallow area of the upper portion of Lake Chesdin provides an excellent environment for algal growth and probably explains the higher standing crop values observed at sites 4 and 5 than at the other sites (Fig. 2). Extremely heavy rains during July returned the lake to the extremely turbid conditions that were present during winter and early spring. In PHYTOPLANKTON SEASONAL CHANGES, LAKE CHESDIN 263 Table V Title: Frequency of Collections (October 1974 - September 1975) Month Numb er of Collections Month Number of Collections October 2 April 2 November 2 May 4 December 1 June 4 January 3 July 4 February 2 August 1 March 2 September 0 n this turbid water, several flagellated species of phytoplankton that had i disappeared during May, 1975, reappeared in large numbers. These species, including Pandorina mo rum , Eudorina elegans , Cryptomonas spp., Synura spinosa , and Mallamonas caudata , could have been very sensitive to high i light intensities, thus preferring turbid water conditions in which light i is reduced. In addition, lowered predation rates by zooplankters could £ have resulted from the high turbidity. Dissolved oxygen concentrations q and water temperature were observed to be factors of less importance for it the flagellated species since these parameters following the heavy rains t varied greatly from observations made during early spring when the annual . maximum in population occurred for these species. Conclusion The phytoplankton of Lake Chesdin is typical of the man-made lakes | of the southeast region of the United States (10). The phytoplankton g; association could be characterized as eutrophic diatom and possibly i results from the availability of high concentrations of important limiting | nutrients and from the morphometry of the lake. The relatively small i drainage area of the Appomattox River above Lake Chesdin allows the lake 3: water temperatures to closely follow the ambient air temperatures and, f thus, normal seasonal changes in the physical environment result in a C corresponding and immediate seasonal variation in total phytoplankton | standing crop. Appendix I Taxonomic List of Lake Chesdin Phytoplankton Observed From October 2, 1974 - August 4, 1976 I (Numbers in right column = month of year observed) * - Diatoms identified after acid clearing I Chlorophyta (152) Class Chlorophceae Order Volvocales (13) Family Chlamydomonadacea Chlamydomonas spp. Ehrenberg Perennial Chlamydomonas pertyi Goroshankin 4,5 Carteria sp. Diesing 7 Family Oocystaceae Zoochlorella conductrix Brandt 7 Chlorella spp. Beyerinck 3, 4, 6-8 Oocystis borgei Snow 6 Oocystis elliptica W. West 5-8 Chodatella subsalsa Lemmermann 6 Franceia ovalis (France) Lemmermann 5 Radiococcus nimbatus (de Wild) Schmindle6 Pachycladon umbrinus G. M. Smith 5-8 Ankistrodesmus braunii (Naeg) Brunnthaler 264 THE VIRGINIA JOURNAL OF SCIENCE Ankistrodesmus convolutus Corda 5 , 7 Ankistrodesmus f alcatus var . mirabilis (West & West) G. S. West 1-4 Ankistrodesmus nannoselene Skuja 1 Closteriopsis longissima var . tropica West & West 5-8 Selenastrum gracile Reinsch 6 Kirchneriella contorta (Schmindle) Bohlin 5 Kirchneriella elongata G. M. Smith 6 Kirchneriella lunaris (Kirchner) Mobius 6 Kirchneriella obesa (W. West) Schmindle 5-7 Quadrigula chodatii (Tanner-Fullman) G. M. Smith 6 Quadrigula lacustr is (Chod.) G.M. Smith 2-8 Tetraedron caudatum (Corda) Hansgirg 7 Tetraedron constrictum G. M. Smith 5, 8 Tetraedron gracile (Reinsch) Hansgirg 5-7 Tetraedron planctonicum G. M. Smith 8 Tetraedron quadricuspidatum (Reinsch) Hansgirg 7 Tetraedron regulare Kutzing 6, 8 Tetraedron trigonum (Naeg.) Hansgirg 5-8 Tetraedron trigonum var. gracile (Reinsch) De Toni 6, 7 Tetraedron trigonum var . setigerum (Arch.) Lemmermann 6, 7 Family Scenedesmaceae Scenedesmus abundans var . brevicauda G. M. Smith 6, 7 Scenedesmus acuminatus (Lag.) Chodat 1, 3, 5-8 Scenedesmus anomalus (G. M. Smith) Ah 1 strom & Tiffany 3 Scenedesmus armatus (Chod.) G. M. Smith 4 Scenedesmus bi juga (Turp.) Lagerheim 5-8 Scenedesmus bi juga var . alternans (Reinsch) Hangirg 6, 7 Scenedesmus denticulatus Lagerheim 6 Scenedesmus denticulatus var . recurvatus Schumacher 5-8 Scenedesmus quadricauda (Turp.) Brebisson 2, 4-8 Scenedesmus quadricauda var. maximus West & West 5,6 Scenedesmus serratus (Corda) Bohlin 4-7 Actinastrum gracilimum G. M. Smith 4 Ac tina strum hantzschii Lagerheim 5, 6 Actinastrum hantzschii var . f luviatile Schroeder 4-7 Crucigenia opiculata (Lemm.) Schmidle 7, 8 Crucigenia irregularis Wille 6, 7 Crucigenia quadrata Morren 6, 10 Crucigenia tetrapedia (Kirch.) West & West 6 Crucigenia truncata G. M. Smith 6 Family Micratiniaceae Micratinium pusillum Fresenius 5-7 Micratinium quadrisetum (Lemm.) G. M. Smith 6-8, 10 Errerella bornheimiensis Conrad 6-8 Family Dictyosphaeriaceae Dictyosphaerium ehrenbergianum Nageli 5, 6 Dictyosphaerium pulchellum Wood 6-8 Order Zygnematales (60) PHYTOPLANKTON SEASONAL CHANGES, LAKE CHESDIN 265 Family Zygnemataceae Mougeotia spp. Agardh 1- 3, 7, 8, 10 Zygnema spp. Agardh 10 Spirogyra spp. Link 1, 3- 6, 10 Family Mesotaeniaceae Cylindrocystis brebissonii Meneghini 8 Family Desmidiaceae Closterium abruptum (Lyngb.) Bresbisson 2 Closterium acerosum (Shrank) Ehrenberg 5- 7 Closterium ehrenbergii Meneghini 5 Closterium gracile Brebisson 3 Closterium gracile var. elongatum West & West 2, 4- 8, 10, i: Closterium gracile var. intermedium Irenne-Marie 4, 6 Closterium incurvam Brebisson 6, 10 Closterium leibleini Kutzing 3 Closterium lineatum Ehrenberg 1 Closterium lunula Nitzsch 7 Closterium moniliferum Brebisson 11 Closterium praelongum Brebisson 5 Closterium praelongum var. brevior West 6 Closterium pronum Brebisson 3 Closterium setaceum Ehrenberg 11 Closterium sublatum (Kutz.) Brebisson 5, 6 Closterium toxon West 1 Closterium turgidum Ehrenberg 3, 4, 6 Pleurotaenium trabecula (Ehren.) Nageli 3, 10 11 Euastrum affine Ralfs 12 Micrasterias denticulata Brebisson 7 Micrasterias radiata Hassall 6, 7, 10 Micrasterias radiata var. dichomata (Wolle) Cushman 7 Micrasterias radiosa var. aculeata Kreiger 7 Micrasterias rotata (Grev.) Ralfs 1 Cosmarium spp. Corda 2, 3, 5 -7, 11 Cosmarium botrytis Meneghini 5, 7 Cosmarium granatum Brebisson 5 Cosmarium minutissimum Archer 6- 8 Cosmarium obtusatum Schmidle 3, 5 Cosmarium ordinatum (Boerg) West & West 7 Cosmarium subcrenatum Hantzsch 5, 7 Cosmarium tenue Archer 6 Arthrodesmus octocornis Ehrenberg 3- 5, 7 Arthrodesmus subulatus Kutzing 7 Staurastrum altenans Brebisson 5, 7 Staurastrum americanum (West & West) G. M. Smith Staurastrum chaetoceras (Shrod.) G. M. Smith 5- 7 Staurastrum curvatum W. West 6 Staurastrum dickei var. maximum West & West 6 Staurastrum dickei var. rhomboideum West & West 7 Staurastrum gracile Ralfs 3 Staurastrum leptocladum var. insigne West & West 7 Staurastrum natator West 10 -12 Staurastrum orbiculare var. depressum Roy & Bliss 266 THE VIRGINIA JOURNAL OF SCIENCE Staurastrum paradoxum var . cingulum Wesi & West Staurastrum pentacerum (Wolle) G. M. Smith Staurastrum punctulatum Brebisson Staurastrum subcruciatum Cooke & Wille Staurastrum tetracerum Ralfs Sphaerozosma granulatum Roy & Bliss Hyalotheca dissiliens (Smith) Brebisson Hyalotheca mucosa (Dillw.) Ehrenberg Desmidium aptogonum Brebisson Desmidium Baileyi (Ralfs) Norstedt Desmidium Swar tzii Agardh Chrysophyta (116) Class Xanthophyceae (6) Order Heterococcales Family Characiopsidaceae Characiopsis cy lindr ica (Lembert) Lemmermann Family Centritractaceae Centritractus dubius Printz Pseudotetraedron neglectum Pascher Ophiocy t ium capitatum var. longispinum (Moebius) Lemmermann Ophiocytium desertum var. minor Prescotl Class Chrysophyceae (20) Order Ochromonadales Family Ochromonadaceae Uroglenopsis americana (Calkins) Lemmermann Anthophysa vegetans (Mull) Stein Family Dinobryonaceae 6 5-8 3, 6 2, 3 2-4, 7 1, 3 1, 4 3 10 4 5, 7 Dinobryon acuminatum Ruttner 3-5 Dinobryon bavaricum Imhof 5-8 Dinobryon cylindricum Imhof 1-8, 10-12 Dinobryon divergens Imhof 5-8, 10, 11 Family Synuraceae Mallomonas acaroides Perty 4-8, 11, 12 Mallomonas apochromatica Conrad 6, 10 Mallomonas caudata Iwanoff 1, 2, 4-8, : Mallomonas producta Iwanoff 6, 7 Mallomonas teilingii Conrad 3 Mallomonas tonsurata Teiling 5 Synura adamsii G. M. Smith 7, 10-12 Synura petersenii Korshikov 5 Synura sphagnicola Korshikov 3-7 Synura spinosa Korshikov 4-8, 10 Synura uvella Ehrenberg 1-4, 7, 11, Order Chromulinales Family Chrysococcaceae Chrysopyxis bipes Stein 6 Order Rhizochrysidales Family Rhizochrysidaeae Rhizochrysis limnetica G. M. Smith 7, 10 Family Stylococcaceae Lagynion reductum Prescott 5-8 Class Bacillariophyceae (90) Order Eupodiscales Family Coscinodiscaceae *Melosira granulata (Ehr.) Ralfs Perennial *Melosira granulata var. angustissima 0. Muller Perennial *Melosira varians Agardh Perennial PHYTOPLANKTON SEASONAL CHANGES, LAKE CHESDIN 267 *Cyclotella stelligera Cl eve & Grunow *Stepheanodiscus hantzschil var . pusilus (Grun.) Kreiger Family Phacotacea Pteronoman sinuosa Chodat Col lomonas orbicularis Stein Family Volvocaceae Gonium pectorale Mueller Pandorina morum Bory Eudorina elegans Ehrenberg Eudorina unicoctta G. M. Smith Volvox aureus Ehrenberg Volvox perglobator Powers Volvox prolif icus Iyengar Family Haematococcaceae Haematococcus lacustris Agardh Order Tetrasporales (3) Family Palmellaceae Sphaerocystis schroeteri Chodat Gloeocystis vesiculosa Naegeli Asterococcus limneticus G. M. Smith Order Ulotrichales (2) Family Chaetophoraceae Draparnaldia sp. Bory Family Protococcaceae Protococcus sp. Agardh Order Oedogoniales (2) Family Oedogoniaceae Oedogonium spp. Link Bulbochaete sp. De Bary Order Chlorococcales (72) Family Chlorococcaceae Golenkinia radiata Chodat Family Characiaceae Characium ambiguum Hermann Characium debaryanum (Reinsch) De Toni Characium pringsheimii A. Braun Characium rostratum Reinhafd Family Hydrodictyaceae Pediastrum biradiatum Meyen Pediastrum boryanum (Turp.) Meneghini Pediastrum duplex Meyen Pediastrum duplex var. gracillimum West 4-8 5 7 7, 8 10, 4-7 1, 3-8, 10 All but 2 5 1-3, 6-8 7 6-8 3, 5 6, 7 7 6-8 5 5 2-7, 10 6 5-7 1, 7 2, 3 2 5 2, 7 3 2-5, 10 6 West 5-8 Pediastrum duplex var. reticulatum Lagerheim 4, 5 Pediastrum duplex var. rotundatum Lucks 7 Pediastrum simplex (Meyen) Lemiuermann 5 Pediastrum tetras (Ehrenb.) Ralfs 5, 6 Pediastrum tetras var. tetraodon (Corda) Rabenhorst 5 Family Coelastraceae Coelastrum cambricum Archer 6 Coelastrum morus West & West 3-8 Coelastrum proboscideum Bohlin 7 Coelastrum sphaericum Nageli 5 Order Rhizosoleniales Family Rhizosoleniaceae Rhizosolenia longiseta Zacharias 6-8 Order Biddulphiales Family Chaetoceraceae Attheya zachariasi Brun. Order Fragilariales Family Tabellariaceae 5-8 268 THE VIRGINIA JOURNAL OF SCIENCE *Tabellaria f enestrata (Lyng.) Kutzing 11 12 *Tabellaria f locculosa (Roth) Kutzing 1 Family Fragilariaceae *Fragilaria brevistr iata Grun . 2-4 *Fragilar ia capucina var . mesolepta Rabh .Perrenial *Fragilaria crotonensis Kit ton 6, 7 *Asterionella f ormosa Hassal 1-7, 11, 12 *Asterionella formosa var . gracillima (Hantz.) Grunow 1-5 *Synedra delicatissima W. Smith 1-8 *Synedra pulchella (Kutz.) Ralfs 2-8 *Synedra pulchella var. lacerata Hust 2-8 *Synedra rumpens Kutz. 1-8 *Synedra ulna (Nitz.) Ehr. Perennial Order Eunotiales Family Eunotiaceae *Eunotia curvata (Kutz.) Lagerheim 3, 4 *Eunotia elegans Oestrup 2-4 *Eunotia pectinalis (Kutz.) Rabenhorst 2, 3, 5-7 A'Eunotia pectinalis var. ventricosa Grunow 5 Eunotia sp. Ehrenberg 5-7 Order Achnanthales Family Achnanthaceae *Cocconeis fluviatilis Wallace 3, 4 *Cocconeis placentula Ehrenberg *Cocconeis placentula var . lineata (Ehr. ) Cleve 2-8 Order Navicuales Family Naviculaceae *Frustulia rhomboides (Ehr.) deToni 2-4 *Frustulia rhomboides var . crass inervia (Breb.) Ross 2-4 Amphiprora ornata Bailey 2-4, 7 *Gyro sigma obtusatum (Sulliv.) Boyer 3, 4 *Gyrosigma scalproides (Rhab.) Cleve 1 Stauroneis acuta W. Smith 2, 3 *Stauroneis anceps Ehrenberg 4 ^'Stauroneis phoenicenteron (Nitz.) Ehrenberg 3 ^'Stauroneis phoenicenteron f . gracilis Hustedt 3 *Neidium affine var. amphirhynchus (Ehr.) Cleve 6, 7 *Nitzschia linearis var. tennis W. Smith 2, 3 *Nitzschia obtusa var . brevissima W. Smith 7 *Nitzschia obtusa var . scalpel 1 if ormis Grunow 2-4 *Nitzschia punctata Grunow 7 'ANitzschia sigmoidea (Ehr.) W. Smith 3-5, 7 *Nitzschia sublinearis Hustedt 3, 4 A'Nitzschia thermalis Kutzing 2-4 Order Surirellales Family Sur irellaceae ~A'Cymatopleura solea (Breb.) W. Smith 2, 3 *Surirella angustata Kutzing 3, 4 *Surirella biseriata var. bifrons (W. Smith) Hustedt 6 A'Surirella elegans Ehrenberg 3-8 *Surirella ovalis Brebisson 3, 4, 6, 7 *Surirella patella var. neupaueri (Panti) Cleve-Euler 2-4 PHYTOPLANKTON SEASONAL CHANGES, LAKE CHESDIN 269 *Surirella robusta var. splendida (Ehr.) van Huerck *Surirella robusta var. tenera (Greg.) van Huerck Cyanophy ta (15) Class Myxophyceae Order Chroococcales Family Chroococcaceae Chroococcus sp. Naegeli Anacystis cyanea Drouet & Dailey Order Hormogonales Family Oscillatoriaceae Spirulina laxa G. M. Smith Spirulina norstedtii Gomont Oscillatoria angustissima West & West Oscillatoria limosa (Roth) Agardh Oscillatoria pr inceps Vaucher Oscillatoria sp. Vaucher Lyngbya birgei G. M. Smith Lyngbya martensiana Meneghini Family Nostocaceae Anabaena af finis Lemmermann Anabaena circinalis Rabenhorst Anabaena limnetica G. M. Smith Aphanizomenon f los-aquae Ralfs Euglenophyta (42 ) Class Euglenophyceae Order Euglenales Family Euglenaceae Euglena acus Ehrenberg Euglena ehrenbergii Klebs Euglena elastica Prescott Euglena minuta Prescott Euglena oxyuris Schmarda Euglena oxyuris var. minor Prescott Euglena proxima Dangeard Euglena spirogyra Ehrenberg Euglena spiroides var. annulata Gojdics Euglena sp. Ehrenberg Phacus brevicaudata (Klebs) Lemmermann Phacus crenulata Prescott Phacus curvicauda Swirenko Phacus helikoides Pochman Phacus lemmermanni (Swir.) Skvortzow Phacus longicauda (Ehr.) Dujardin Phacus norstedtii Lemmermann Phacus orbicularis Hubner Phacus pleuronectes (OFM) Dujardin Phacus suecius Lemmermann Phacus triqueter Playfair Lepocinclis acicularis France Lepocinclis acuta Prescott Lepocinclis ovum (Ehr . ) Lemmermann Lepocinclis texta (Duj . ) Lemmermann Trachelomonas acanthophora Stokes Trachelomonas armata var . longispina (Playf.) DeFlandre Trachelomonas armata var . steinii Lemmermann Trachelomonas gibberosa Playfair Trachelomonas hispida (Perty) Stein Trachelomonas hispida var . coronata Lemmermann 2, 3 3-8, 10 5-8, 10, 12 6 5, 6 7 5-7, 12 6 5, 6, 11 1, 5-7, 10, 11 5 10 5-7 5-8 1, 5-8, 10-12 Perennial 6 5-8 5-8 5-7 1-3, 5-7 1, 3-8 7 3-8, 10 All but 12 3-5, 10 5 5 7 2, 5, 6 3-7, 10, 11 6, 7 5- 8 7 5 7, 8 5, 7 5 6- 8 5, 7 6, 7 5, 7 7 2, 6, 7 Perennial 270 THE VIRGINIA JOURNAL OF SCIENCE Trachelomonas hispida var. crenulatorollis (Mask. ) Leiraneraann 4-6 Trachelomonas horrida Palmer 1, 3, 4, 6-8 Trachelomonas kelloggii (Skv.) DeFlandre5 Trachelomonas playf airii DeFlandre 1 Trachelomonas robusta Swirenko 7 Trachelomonas scabra var . longicollis Playfair Trachelomonas schauinslandii Lemmermann 3, 5-7 Trachelomonas superba (Swir.) DeFlandre 2, 5-7 Trachelomonas sydneyensis Playfair 7 Trachelomonas volvocina Ehrenberg All but 1 Trachelomonas zmiewiki Swirenko 2-5 Pyrrhophy ta (12) Class Dinophyceae Order Gymnodiniales Family Gymnodiniaceae Gymnodinium fuscum (Ehr.) Stein 2, 5 Gymnodinium palustre Schilling 6 Gymnodinium limnetica Lackey 7 *Neidium bisulcatum (Lagerst.) Cleve 7 *Neidium iridis var. amp lia turn (Ehr.) Cleve 7 *Diploneis f innica (Ehr.) Cleve 2 *Navicula cuspidata Kutzing 3, 5 *Navicula cuspidata var. major Meist. 2-4 *Navicula exigua var. capitata (Grun.) Greg. _ 3 *Navicula mobiliensis var. minor Patrick 2-4 *Navicula radiosa Kutzing 2-4 *Navicula rhynchocephala Kutzing 7 Navicula spp. Bory. Perennial *Pinnularia biceps Gregory 3-6 *Pinnularia legumen Ehrenberg 3 , 4 *Pinnularia microstauron (Ehr.) Cleve 2-4 *Pinnularia viridis (Nitz.) Hustedt 2-7 "Pinnularia vir id is var . commutata (Grun.) Cleve 3, 4 *Gomphonema acuminatum Ehrenberg 1-7, 12 "'Gomphonema acuminatum var . coronatum (Ehr.) W. Smith 2-4 *Gomphonema auger Ehrenberg 7 *Gomphonema constrictum var . capitatum (Ehr.) Cleve 2-8 "'Gomphonema gracile Ehrenberg 2-4 *Gomphonema parvulum Kutzing 5 *Gomphonema parvulum var . lanceolata Kutzing 3-5 *Caloneis lewisii Patrick 2, 6 "Cal one is limosa (Kutz.) Patrick 3, 7 Family Cymbellaceae 'fCymbella af finis Kutzing 7 "Cymbella cistula Hempr . *Cymbella cuspidata var . naviculif ormis Auersw . "Cymbella cymbiformis Ehrenberg 3 , 4 ^Cymbella ehrenbergii Kutzing 1”3, 5-8 "Cymbella gastroides Kutzing 3, 6 *Cymbe 11a lanceolata (Ehr.) Van Huerck Perennial "Cymbella obtusa Gregory 6 "Cymbella tumida (Breb.) van Huerck 6, 7 *Cymbella turgida (Gregory) Cleve 3-5 "'Amphora ovalis f. minor Kutzing 3, 5, 7 PHYTOPLANKTON SEASONAL CHANGES, LAKE CHESDIN 271 Order Epithemiales Family Epithemiaceae *Epithemia turgida (Ehr.) Kutzing 1-3 , “Rhopalodia gibba (Ehr,) Muller 1, 2, *Rhopalodia g ibba var . ventricosa (Ehr.) Grunow 6, 7 Order Nitzschiales Family Nitzschiaceae *Nitzschia f asiculata Grunow 3 , 7 Order Peridinales Family Glenodiniaceae Glenodinium sp. (Ehr.) Stein 6-8 Glenodinium quadridens (Stein) Schiller 7, Family Per idiniaceae Per idinium aciculiferum Lemmermann 6, Per idinium cine turn (Muell.) Ehrenberg 5-8 Peridinium cinctum var . tuberosum (Meunier ) Lindemann 5 Peridinium limb a turn (Stokes) Lemmermann 6 Peridinium pus ilium (Penard) Lemmermann 4-6 Peridinium wisconsinense Eddy 7 Family Ceratiaceae Ceratium hirundinella (Muell.) Dujardin 6-8 5-8 4-8 7 , 1 Cryptophyta (2) Class Cryptophyceae Order Cryptomonadales Family Cryptomonaceae Cryptomonas erosa Ehrenberg Perennial Cryptomonas spp. Ehrenberg Perennial References 1 . Anonymous . Standard Me thods for the Examination of Water and Wastewater. Am. Public Health Association, American Water Works Association, Water Pollution Control Federation, 12th Edition, 1965. 2. Palmer, C. M. Evaluation of New Algicides for Water Supply Purposes. Taste and Odor Control Journal 23(1), 1-4, 1957. 3. Gibson, C. E. The Algicidal Effect of Copper on a green and Blue-green and Some Ecological Implications. Journal of Applied Ecology 9 (2), 513-518, 1972. 4. Whitford, L. A. Phytoplankton in North Carolina Lakes and Ponds. Journal of the Elisha Mitchell Society 74(2), 143-157 (1958). 5. Poore, W. H. and H. G. Marshall. Lake Drummond of the Dismal Swamp: I. Phytoplankton Composition, Virginia Journal of Science 23, 72-76, 1972. 6. Obeng-Asamoa , E. K. and B. C. Parker. Seasonal Changes in Phytoplankton and Water Chemistry of Mountain Lake, Virginia Transactions of the American Microscopical Society 91(3), 363-380, 1972. 7. Hutchinson, G. E. A Treatise on Limnology, Vol. II - Introduction to Lake Biology and the Limnoplankton , John Wiley and Sons, New York, 1967. 8. Anonymous. Lake Chesdin Preliminary Report, Environmental Protection Agency, National Eutrophication Survey, Corvallis, Oregon. 1975. 272 THE VIRGINIA JOURNAL OF SCIENCE 9. Whetstone, G. W. and R. L. McAvoy. Chemical Quality of Surface Waters of Virginia, 1946-48. Bulletin No. 11. Division of Water Resources, Charlottesville, Virginia. 1948. 10. Pearsall, Witt. Phytoplankton in the English Lakes, II The Composition of the Phytoplankton in Relation to Dissolved Substances. Journal of Ecology 20, 241-262, 1932. Virginia Journal of Science Volume 34, Number 4 Winter, 1983 Federalism and its Impact on NSF Grants and the Training of Teachers7 Ertle Thompson Past President, Virginia Academy of Science Treasurer, National Association of Academies of Science Professor of Science Education Ruffner Hall Charlottesville, Virginia 22903 ABSTRACT The greatest challenge for science and mathematics education in the ’80’s is the establishment of realistic goals for developing a scientifically literate society, and the preparation and enhancement of the talent pool for the research and development, and the technological support essential for human welfare in a world increasingly dependent on science and technology. Recent studies and surveys indicate declining knowledge and interest in science and mathematics among pre-college students; yet, the federal govern¬ ment and, in many cases, state governments have abandoned the aggressive efforts of the post-Sputnik era to improve the quality of science and mathematics education. Teacher preparation suffers in both quality and quantity. Science and mathematics education centers are being devastated by retrenchment policies. Instructional materials and sup¬ plies are inadequate. Teachers are leaving the classroom for more financially rewarding careers. In-service-continuing education opportunities to upgrade teacher knowledge and keep teachers alive professionally have been drastically reduced. Our challenge is to modify the policies and practices responsible for the present status, so that science, mathematics and technological education for the ’80’s will be strengthened. This paper is dedicated to the thousands of science teachers who daily perform the task of teaching the youth of this country in high quality programs in science education. Despite the continuing criticisms in the news media and public meetings, and inadequate support, they continue to produce from among our top students the best scientific talent in the world. INTRODUCTION AND OVERVIEW The annals of science education literature are replete with yearbooks and commission studies analyzing and delineating the problems of science education for the past hundred 'A presentation given to the 1983 NAAS/ A A AS Symposium: The Crises in Science and Mathematics Education. 274 THE VIRGINIA JOURNAL OF SCIENCE years. The collection in my office currently occupies 12 linear feet of shelf space. All of you present today are familiar with most of the yearbook and commission studies published during the past two decades; therefore, it is the choice of the writer not to bore you with the redundancy of the commission findings and recommendations. The writer would, however, like to emphasize the tremendous change that has come about in American education since the publication of the Committee of Ten Report in 1893, which recommended a highly content structured program of preparation in science for the 8 to 11 percent of the 14-17 year old school population who attended secondary schools at that time. These recommendations were made despite the fact that only 2 per¬ cent of the eligible school age population actually graduated from high school. These data need to be compared and contrasted with the recommendations concerning the cur¬ riculum development and implementation of the 60’s and 70’s when 90 percent of the school age population were in school and 75 percent of the eligible population graduated from high school. Obviously, the demands on the schools and upon the teachers of specialized areas, such as science and mathematics, are much more diverse in the 1980’s than the 1890’s. Yet one could easily deduce, from recent recommendations for a return to the rigors of the past, that many feel the academic program recommended by the Committee of Ten would suit the youth of today. CHANGING ORGANIZATIONAL PATTERNS OF SCHOOLS AND CONFLICTING DEMANDS OF THE PUBLIC It is probably safe to assert that no other educational system in the world has at¬ tempted to do as much for so many as we have in the 50 states of this great nation. By attempting to educate all of the children of all of the people we have increasingly placed our teachers in situations requiring them to teach not only the content of the academic discipline in which they should be prepared, but to deal with an ever increasing number of social issues from a complex and ever changing society. The awareness of social in¬ justices of the past and the desire to solve the problems and injustices created by discriminatory actions have also increased demands on teacher’s time and commitment. In the midst of this school climate, and increasing awareness of the need for protection of individual student rights and freedom, classroom standards for control and self- discipline became very much confused. Standards for student performance deteriorated. The public cry for accountability sounded louder and louder until “Back to Basics” became the marching theme from coast to coast. Problems — Challenges — Crises? Several important areas to be discussed in the remaining section of this paper deal with the problems and challenges confronting the science education community, which if not attended to will certainly become crises. Goals for Science Education We need to address ourselves, in the science education community, to agree upon two realistic and attainable goals. (1) The teaching of science for development of the scientific talent in the academic disciplines for future careers in science, mathemtics, engineering, and technology and (2) a commitment to the development of scientific literacy in the total population, for fuller understanding and enjoyment of the world of nature and technology, and wiser decision making regarding the problems of daily living in the FEDERALISM’S IMPACT ON TEACHER TRAINING 275 technological society of today and tomorrow. The curriculum development efforts post-Sputnik under the support of the National Science Foundation funding, and quality control by specialists and teachers, had the most significant impact of any other individual effort in the history of American educa¬ tion. Despite much data accumulated in the national surveys showing that the cur¬ riculum projects fell short of their stated goals, it certainly cannot be argued that the cur¬ riculum projects failed to infuse the schools K-12 with up-to-date science content. In retrospect, many of the courses and programs may have been overly structured in terms of the academic discipline at the expense of eliminating most or all of the technology, which for many provided the interest and motivation for the study of the applications of i science. The major failures of the curriculum project movement of the 60’s and 70*s can be categorized as the implementation activities, or lack thereof. The curriculum develop¬ ment and implementation phases were not buttressed by teacher preparation programs, conducted by knowledgeable, enthusiastic, dedicated individuals. This is not to say that the support for summer institutes, academic year institutes, and in-service teacher preparation programs was inadequate, but simply to assert that because of the lack of coordination, the left hand did not know what the right hand was doing, and the teacher preparation activities, in many cases, provided negative support for the cur¬ riculum development products and implementation efforts. Support Mechanisms A continuing, perplexing problem of American education is the lack of clearly defined roles for the federal, state, and local governments in the financial support of education. 1 The disparity among the financial bases of local communities at the state level and among states at the national level means that a local community within a state may ex¬ pend a tenfold effort when compared to another local community within the state and still provide inadequate financial support for needed educational services. The same is true when the poorest state is compared to the wealthiest state in terms of tax base and ef¬ fort. Such inequities must be addressed individually by the states and also by the federal government. During the post-World War II era, communities provided the financial support for the construction of school facilities unequalled in our history. However, with increasing in- I flation, the rising cost of living and higher taxes the public revolt of the late 70’s left public education at the mercies of supercritical and unsupportive electorates. Teachers were expected to perform miracles in instructional programs with science, where thetotal ' equipment and instructional supplies budget per pupil annually was less than the cost of a ticket, with refreshments, for a NFL football game. Teachers’ salaries were inadequate | and the annual increases were zero to a fractional part of the inflation rate. The Abdication of Responsibility for Teacher Preparation In the mid-50’s and the post-Sputnik era science education in America, and inter¬ nationally, witnessed a great resurgence of cooperative efforts on the part of scientists, engineers, technicians, and educators under the auspices of National Science Founda¬ tion support. College and university professors in all of the major disciplines of science *1 devoted themselves to special programs for teacher preparation, the undergraduate and graduate continuing education. However, with much of the conflict surrounding con- | cems for social issues in the 70’s and the decline in federal financial support for science and mathematics education activities most of the academic scholars retreated to their research laboratories, where they could be safe from public ridicule, and more secure in 1 the financial support of continuing research grants. 276 THE VIRGINIA JOURNAL OF SCIENCE Much of the preceding discussion is too negative, and too candid an indictment of the scientific and education communities of this country. However, if we are to solve the problems of the future we must be willing to share the responsibilities for our individual actions and the blame for our failures as readily as we are willing to accept the accolades for our achievements when we succeed. Fortunately there are many positive indicators which should serve as guides in the search for solutions to our current problems. The public seems receptive to massive changes in educational programs. The states are redefining standards for instructional programs, and mathematics and science require¬ ments are being increased nationwide. Departments of teacher certification are establishing higher standards for the certification of science teachers. Colleges and universities are making concerted efforts to enroll more teachers in undergraduate pro¬ grams leading to certification in the sciences and mathematics. The challenge remains, from the White House and the Halls of Congress to the local school board and the principal’s office, for executive, legislative, and administrative policies and financial support for excellence in instructional programs. The same is true, from the President’s Science Advisor and the National Science Board to the college pro¬ fessor, to the local science supervisor, regarding responsibilities for establishment of pro¬ fessional policies and support which have academic merit and credibility. The prepara¬ tion of science teachers for the future is too important and too massive an undertaking to be left to the discretion of a small cadre of science education specialists in far too limited number of colleges and universities. It must be the responsibility of the whole scientific community. Past and Future Roles of NSF Grants and the Training of Teachers Little has been stated explicitly, but much has been implied in the preceding sections of this paper regarding the impact of the new Federalism on NSF’s participation in the training of science and mathematics teachers. However, from reading the myriad of reports and studies from the inception of the National Science Foundation to the present it is not clear that all of the reduction in NSF participation in science and mathematics education activities at the pre-college level have been due to the new Federalism. From a very meager beginning in 1952, with an expendityre of $7,000 plus for science education research and analysis the support and involvement of NSF grew to a maximum SI 24 million for each of the years 1966-68, and then diminished to practically zero in 1983. A graph of the expenditures during that period of time represents an almost perfect Bell curve. The support initially for teacher education in Summer Institutes, then Academic Year Institutes, followed by Course Content Development Projects in the disciplines and implementation activities including specialized teacher training involved estimates of 50-60 percent of the mathematics and science teachers grades 7-12. During the peak period of financial support and teacher participation it is estimated that as many as 30 percent of the teachers in mathematics and science were provided the best of training, and/or continuing education in combinations of Summer Institutes, In-service In¬ stitutes, and Cooperative College School Science Programs. The role of NSF was clearly that of providing, as authorized by the National Science Foundation Act of 1950, “financial support for the scientific and educational activities and the appaisal of the im¬ pact of such activities upon the general welfare.” The secondary role of NSF, unrecognized by many in the scientific community, was the establishment and maintenance of standards through the peer-review system for programs in teacher train¬ ing at the graduate, and continuing education levels. The established standards directly FEDERALISM’S IMPACT ON TEACHER TRAINING 277 effected positive changes in undergraduate teacher education programs. With the precipitous decline in funding of the Science Education Directorate within the National Science Foundation, beginning in 1969, the role that NSF could play, and has played in helping to establish and maintain standards of excellence in science and mathematics education at the pre-college level diminished significantly. It is the considered judgment of this writer that the future role of NSF must be to help reestablish those standards by the provision of the scholarly expertise, and serving as the major conduit of Financial support for redefined and reestablished programs of excellence in science, mathematics, engineering, and technological education. To reestablish and redefine its role NSF must have the support from the new generation of young scientists, mathematicians, engineers, and teachers who constitute the membership of the scientific societies and associations, such as the National Association of Academies of Science and the Amer¬ ican Association for the Advancement of Science. With the concerted and cooperative efforts of all such interestsed persons the declines of the past ten years can be reversed and sound programs of teacher preparation for the future can be reestablished. 278 Requested Amendment to ByLaws Proposed Bylaw changes must be distributed to the membership or published in the Virginia Journal of Science at least 30 days prior to action by Council. Adoption reguires an affirmative vote of a majority of the t otal membership of Council. Action on the proposed changes presented here is planned for the March Council meeting. The Nominations and Elections Committee and Council have recommended that the following amendment to the Bylaws be processed; Section 10. Nominations and Elections Committee shall: (1) Mail to the membership on or about January 1 each year a request for nominations of persons to fill the offices of President-elect, Secretary and Treasurer. * (2) Nominate a slate of two persons for each of the aforenamed offices and present to March Council Meeting. (3) Mail slate of nominees to members advising that names may be added to the slate by 25 members petitioning the committee on behalf of each name to be added. (4) Prepare ballots with or without additional nominees as the case may be and mail to membership with registration and other information relative to annual meeting indicating deadline and address for return of ballot to committee. (5) Count ballots and announce results at the Academy Conference. Should a tie vote result for any office, the Academy Conference shall vote on the nominees. In all cases, the nominee receiving the largest number of favorable votes shall be elected; provided, however, that only members in good standing may cast ballots. * Amend paragraph (2) by striking "present to March Council Meeting" and substituting "report to Council for informational purposes." 279 Jeffress Research Grant Awards The Allocations Committee of the Thomas F. and Kate Miller Jeffress Memorial Trust has announced the award of Jeffress Research Grants to the institutions listed below to support the research of the investigator whose name is given. The Jeffress Trust, established in 1981 under the will of Mr. Robert M. Jeffress, a business executive and philanthropist of Richmond, supports research in chemical, medical or other natural sciences through grants to non-profit education and research institutions in the Commonwealth of Virginia. The Jeffress Research Grants being announced have been awarded in the period from February 1982 through May 1983. The Jeffress Memorial Trust is administered by First & Merchants National Bank of Richmond. Additional information about the program of the Trust may be obtained by writing the Advisor, Thomas F. and Kate Miller Jeffress Memorial Trust, Trust Department, First & Merchants National Bank, P.0. Box 26903, Richmond, Virginia 23261. Bruce M. Anderson, Virginia Tech. Mechanistic studies of 20-A-hydroxysteroid dehydrogenase. $49,374 (three years) Bulent Atalay, Mary Washington Colleye. Pertubation Theory for Projected States. $5,200 (two years) Robert K. Boggess, Radford College. The Synthesis, Electrochemical, and Luminescence Investigation of Ruthenium (II) Polypyridine Complexes and Other Studies of Pyridine Ligands. $26,800 (two years) Kang Cheng, University of Virginia. Mechanism of action of insulin: study of existence of a cell membrane protease signalling factor. $60,000 (two years) Michael J. Cloutier, Virginia Tech. Regulation of cellular differentiation by differential gene expression. $13,580 (two years) Donald G. Cochran, Virginia Tech. Uric acid excretion in the woods cockroach, Parcoblatta fulvescens. $14,965 (two years) Mark S. Conradi, The College of William and Mary. NMR and Dielectric Studies of Quadrupolar Glasses. $42,690 (three years) Cecil S. Cummins, Virginia Tech. A proposal to investigate the structure of the cell wall polysaccharides in propioni-bacteri a. $75,486 (three years) Gary C. Defotis, The College of William and Mary. Experimental Studies of Critical Behavior and Phase Diagrams in Dilute and Mixed Magnetic Systems. $38,683 (three years) J. B. Delos and S.K. Knudson, The College of William and Mary. Classical and Quantum Mechanics of Atoms in Strong Magnetic Fields. $38,788 (two years) Mark Fail la, Virginia Tech. Accumlation of copper in the kidney of the streptozotocin diabetic rat: location and influence of the metal on tissue function. $32,633 (two years) Robert M. Grainger, University of Virginia. Control of developmental pathways in embryonic tissues. $98,874 (three years) 280 Stephen Goldberg and J. Ross McClung, Virginia Commonwealth University. Structure-function relationships in extraocular motor units. $104,290 (three years) Brian E. Hanson, Virginia Tech. Investigation of Catalytic Materials by Dynamic High Resolution 31P NMR Spectroscopy. $51 ,345 (three years) W. John Hayden, University of Richmond. Comparative stem ontogeny and phyllome anatomy in Chamaesyce and Euphorbi a subgenus 7\ga1oma . $10,893 (two years) Alan G. Heath, Virginia Tech. Development of a way to measure energy status and growth of a population of fish under stress. $5 ,645 (one year) P. Jena, Virginia Commonwealth University. Electronic Structure of Defect Complexes. $40,500 (three years) Robert A. Jordan, Hampton Institute. Mi croecologi cal factors associated with the occurrence of Acanthamoeba species (sarcodinid protozoans). $65,000 (one year) Bruce L. King, Randol ph-Macon College. Flavonoid systematics of Lychnophora ( vernonieae: compositae) . $24,800 (two years) Kenneth R. Lawless, University of Virginia. High Resolution and Analytical Electron Microscopy Studies of Selected Catalysts. $87,695 (three years) Joseph P. Liberti, Virginia Commonwealth University. Translational control of collagen biosynthesis by somatomedin. $56,340 (two years) Burton J. Litman, University of Virginia. Research projects for an automatic recording spectropol arimeter. $46,167 (one year) Robert E. Ludt, Virginia Military Institute. Heteroatom Directed Lithiation of Aromatic Compounds. $20,175 (three years) Francis L. Macrina, Virginia Commonwealth University. Genetics of antibiotic resistance in bacteriodes. $64,144 (three years) Orson K. Miller, Virginia Tech. A study of the higher fungi of Virginia. $12,500 (three years) Larry Nichter, University of Virginia. Improving longterm patency in mi crovascul ar autogenous and synthetic grafts. $8,600 (one year) . Walter G. Niehaus, Virginia Tech. Mechanisms of regulation of fungal secondary metabolism. $67,316 (three years) Blaine E. Norum, University of Virginia. Electro-nuclear Studies Using the NIKHEF-K Medium Energy Electron Accelerator. $83,300 (three years) John L. Patterson, Jr., Virginia Commonwealth University. Spectral analysis of respiratory sound. $79,049 (three years) S. J. Poon, University of Virginia. Experimental Investigations of Magnetic Interactions and Non-kondo Resistivity Anomalies in Metallic Glasses. $34,340 (two years) 281 Jonathan Ravdin, University of Virginia. Entamoeba histolytica: mechanisms and prevention of amebic adherence and cytolysi s . $86,215 (three years) Lionel I. Rebhun, University of Viryinia. Investigations of multiple forms of tublin in dividing and non-dividing cells. $103,400 (three years) William S. Shear, Hampden-Sydney College. Study of fossils of terrestrial arthropods from the Devonian of Gilboa, New York. $31,000 (one year) ; Peter J. Sims, University of Virginia. A biophysical study of the interaction of complement proteins C5B-9 with lipid containing membranes. $82,161 (three years) Barry E. Stein, Virginia Cormionwealth University. Superior collicus and pain. $146,842 (three years) James A. Stevenson, Virginia Commonwealth University. Optic axon growth from peripheral nerve grafts. $23,470 (two years) | Ronald P. Taylor, University of Virginia. Characterization of pathogenic. DNA-antibody complexes. $6,500 (one year) J James Terner, Virginia Commonwealth University. Rapid Mixiny j Resonance Raman Spectroscopy of Peroxidase and Catalase Inter- I mediates. $59,360 (three years) i David W. Thompson, The College of William and Mary. Organic | Synthesis through Early Transition Metal Organometal 1 ic Chemistry. $30,150 (three years) Stephen T. Thornton, University of Virginia. Mass Measurements of Exotic Nuclei. $37,000 (three years) Wayne H. Tinnell, Longwood College. Genetic transfer in oral anaerobes. $5,400 (one year) Richard W. Topham, University of Richmond. The physioloyical significance of a serum ferroxidase inhibitor. $62,916 (two years) David W. Towle, University of Richmond. An investigation of membrane function in ion-transporting cells of blue crab gill. $46,975 (three years) W. Peter Trower, Virginia Tech. Four Exploratory Research Projects in Physi-cal Science. $36,000 (three years) Bruce J. Turner, Virginia Tech. Mitochondrial DNA as new tool for probing the evolutionary biology and genetic structure of fish populations. $12,230 (one year) Bart Van't Riet, Virginia Commonwealth University. Antitumor Agents Containing Polyhydroxphenyl Rings. $38,400 (three years) Bruce Wallace, Virginia Tech. The genetic assimilation of a heat-shock-induced phenotype in Drosophila mel anogaster . $2,360 (two years) R. C. Whisonant, Radford College. Fabric and orgin of intraclasts in Some Cambro-Ordovici an carbonates. Southwestern Virginia. | $21,470 (two years) 282 Robert H. White, Virginia Tech. Biosynthesis of coenzyme M in methanogenic bacteria. $12,970 (two years) John L. Wiktorowicz, Virginia Tech. Molecular biology of globin gene expression in human cultured cells. $25,486 (one year) A. A. Yousten, Virginia Tech. An investigation of baci 1 1 us laterosporus: A bacterium causing disease in mosquito larvae. $12,525 (two years) 283 Mountain Lake Biological Station Announcement ANNOUNCING MOUNTAIN LAKE BIOLOGICAL STATION SUMMER COURSES 1984 First Term (June 11 - July 14) Herpetology Ronn Altig, Mississippi State University Ornithology Val Nolan, Jr., Indiana University Ellen Ketterson, Indiana University Plant Ecology Elizabeth Lacey, University of North Caro lina-Greens boro Quantitative Methods in Field Biology Joseph Travis, Florida State University Workshop in Alio zyme Techniques Charles R. Werth, College of William and Mary Second Term (July 16 - August 18) Biology of Insects George W. Byers, University of Kansas Community Ecology Norman Christensen, Duke University Henry Wilbur, Duke University Freshwater Ecology Daniel J. Hornbach, University of Virginia Natural History of the Southern Appalachians Robert C. Simpson, Lord Fairfax Community College Nature Photography Robert C. Simpson, Lord Fairfax Community College Workshop in Alio zyme Techniques Charles R. Werth, College of William and Mary Scholarships Available: Service Awards for students covering room and board costs. Post-Doctoral Research Awards (10 weeks) For further information and application write to : Dr. Jerry 0. Wolff, Director Mountain Lake Biological Station Gilmer Hall University of Virginia Charlottesville, VA 22901 NOTES 1 A. TITLE OF PUBLICATION IB. PUBLICATION NO. 2. DATE OF FILING The Virginia Journal of Science 6 6 0 5 8 0 Sept. 23, 1983 U.S. Postal Service STATEMENT OF OWNERSHIP, MANAGEMENT AND CIRCULATION Required by 39 U.S.C. 3685) 3. FREQUENCY OF ISSUE Quarterly-Spring, Summer, Winter, Fall 3A. NO. OF ISSUES PUBLISHED ANNUALLY 3B. ANNUAL SUBSCRIPTION PRICE $27.50 4. COMPLETE MAILING ADDRESS OF KNOWN OFFICE OF PUBLICATION (Street, City, County, State and ZIP Code) (Not printers) Virginia Academy of Science, Dept, of Biology, Univ. of Richmond, Richmond, Va. , USA 23173 5. COMPLETE MAILING ADDRESS OF THE HEADQUARTERS OF GENERAL BUSINESS OFFICES OF THE PUBLISHER (Not printer) SAME AS ITEM 4 6. FULL NAMES AND COMPLETE MAILING ADDRESS OF PUBLISHER, EDITOR, AND MANAGING EDITOR (This item MUST NOT be blank) PUBLISHER (Name and Complete Mailing Address ) Virginia Academy of Science, Dept, of Biology, Univ. of Richmond, Richmond, Va., USA 23173 EDITOR (Name and Complete Mailing Address) Dr. Stewart A. Ware, Department of Biology, College of William and Mary Williamsburg, Virginia 23185 MANAGING EDITOR (Name and Complete Mailing Address ) Dr. Norman J. Fashing, Department of Biology, College of William and Mary Williamsburg, Virginia 23185 7. OWNER (If owned by a corporation, its name and address must be stated and also immediately thereunder the names and addresses of stockholders owning or holding I percent or more of total amount of stock. If not owned by a corporation, the names and addresses of the individual owners must be given. If owned by a partnership or other unincorporated firm, its name and address, as well as that of each individual must be given. If the publica¬ tion is published by a nonprofit organization, its name and address must be stated ) (Item must be completed. ) FULL NAME COMPLETE MAILING ADDRESS Virginia Academy of Science Department of Biology University of Richmond Richmond. Virginia 23173 KNOWN BONDHOLDERS, MORTGAGEES. AND OTHER SECURJTY HOLDERS OWNING OR HOLDING 1 PERCENT OR MORE OF TOTAL AMOUNT OF BONDS, MORTGAGES OR OTHER SECURITIES (If there are none, so state) FULL NAME COMPLETE MAILING ADDRESS NONE 9. FOR COMPLETION BY NONPROFIT ORGANIZATIONS AUTHORIZED TO MAIL AT SPECIAL RATES (Section 423.1 2 DMM only) The purpose, function, and nonprofit status of this organization and the exempt status for Federal income tax purposes (Check one) S HAS NOT CHANGED DURING PRECEDING 12 MONTHS □ HAS CHANGED DURING PRECEDING 12 MONTHS (If changed, publisher must submit explanation of change with this statement. ) EXTENT AND NATURE OF CIRCULATION AVERAGE NO. COPIES EACH ISSUE DURING PRECEDING 12 MONTHS ACTUAL NO. COPIES OF SINGLE ISSUE PUBLISHED NEAREST TO FILING DATE A. TOTAL NO. COPIES (Net Press Run) 2000 May 24, 1983 2000 B. PAID CIRCULATION 1 . Sales through dealers and carriers, street vendors and counter sales 2. Mail Subscription 1388 1420 C. TOTAL PAID CIRCULATION (Sum of I0B1 and I0B2) 1388 1420 D. FREE DISTRIBUTION BY MAIL, CARRIER OR OTHER MEANS SAMPLES, COMPLIMENTARY, AND OTHER FREE COPIES E. TOTAL DISTRIBUTION (Sum of C and D) 1388 1420 F. COPIES NOT DISTRIBUTED 1. Office use, left over, unaccounted, spoiled after printing 612 580 2. Return from News Agents G. TOTAL ( Sum of E, FI and 2— should equal net press run shown in A ) I certify that the statements made by me above are correct and complete 2000 2000 July 1982 S^ifor^ SsS ^3Wl^ ^'CUU^ «#mv? &3& *\lW S% ^ m t*£ ^vcutr^ >'cuu^ ttyis 2S OU^j WjUgl >'CUL7(/^ >HH#g 4yjwl4^ ^KUttyS ($K» *&* ^wcuir^ IS»flnjrf& :'!*JWl^ 2»niov*^ s^JS ^jcutr^; *\» s*te >KUIT(^ (ggsRj gsa%/ V' v^>2in3^ ^'OBM-V/o A'iWU'^ ^KWiO^'V’ jcr