ILLINOIS STATE ACADEMY OF SCIENCE

VOLUME 105 (2012) #1&2, pp. 1-76

f

<

{

5\> ►

1

^uiraiQ

<ts-: U

'M

Illinois State Academy of Science Founded 1907

Affiliated with the Illinois State Museum ISSN 0019-2252

Printed by authority of the State of Illinois Springfield, Illinois

~stat*t

\m

1868 ' 18T8W

luV

86"

ml

L.UCLO 5 S’ » » • tviCf\ I

LIBRARY

ALSO 0 3 2012

F..W — ' " - - INFORMATION FOR CONTRIBUTORS

Submission Information

1. Send original communications and manuscript submissions to: Dr. Tere North, Transactions Editor; via email to terenl956@gmail.com or via postal mail to 1713 W. Adams Rd., Macomb IL 61455.

2. Three (3) hard copies are required.

3. Submission of a manuscript to the Editor will be held to imply that it has not been previously published except in Abstract form and, if accepted, will not be published elsewhere in the same form without written consent of the Editor.

4. Manuscript format should follow a form appropriate to the specific discipline: most commonly, e.g.. Abstract, Introduction, Materials and Methods, Results, Discussion, and Summary.

5. Tables and figures must each be numbered consecutively in the order they are referred to in the text. At manuscript submission, authors may include tables/figures either at the end of or in the body of the manuscript, depending on author preference for publication format.

6. Genus and species must be given for all organisms used in the investigation. Common names are also recommended.

7. In the text, references should be quoted by the author’s name and date, e.g., (Smith, 1960). Footnotes are to be avoided.

8. At the end of the manuscript, full references, must be listed in alphabetical order and include the names of the authors, date of publication, full title of the paper, title of the journal, volume number, and page numbers. References to books must include the number of the edition, publisher, and place of publication.

9. Authors are encouraged to submit names and addresses of potential content reviewers.

Acceptance Information

10. Each manuscript will be reviewed by no less than two anonymous reviewers.

11. For manuscripts deemed acceptable for publication, authors must address the reviewers comments and provide a revised hard copy to the Editor.

Publication Information

12. Upon final acceptance of a manuscript for publication, the author shall provide the Editor with electronic copy (e.g., floppy, CD-ROM, or electronic e-mail attachment) of the final version of the manuscript including all text and tables.

13. All graphs, charts, and diagrams should likewise be provided in suitable electronic format (JPEG, TIFF, PICT, etc.), with a minimum 300 dpi resolution for images, keeping in mind that final printed copy must fit within an 8”x5” area which may include multiple tables/figures per page as long as the appearance of the page is not cluttered. For copy not available electronically, high quality, glossy originals must be provided for direct scanning. Note that layout space must be provided for the Editor to format the Figure descriptions for printing.

14. Galley proofs will be sent to the author or designee for review prior to publication. Corrections to the page proofs are restricted to minor revision and due to printer error. Other alterations will be made at cost and charged to the author.

15. Academy member page charges are $50.00 per page for grey-scale publication. The charge for non-members is doubled. An electronic PDF copy will provided at no additional cost by the Academy following receipt of page charges and publication. At this time, credit card payment is not available for manuscript publication.

Note: Transactions is pursuing expanded online publishing efforts beginning 2013 to enable more rapid publication of author’s manuscripts to the prevent further cost increases.

Transactions of the Illinois State Academy of Science (2012) Volume 105, #1&2, pp. 1-10

received 3/21/11 accepted 1/30/12

The Effect of Nitrogen Level and Form on the Growth and Development of Vetiver Grass ( Chrysopogon zizanioides)

Suneetha Mondyagu1, David E. Kopsell1*, Richard W. Steffen1, Dean A. Kopsell2, and

Robert L. Rhykerd1

1 Department of Agriculture, Illinois State University, Normal, IL 2 Plant Sciences Department, The University of Tennessee, Knoxville, TN ^Corresponding author email: dkopsell@ilstu.edu

ABSTRACT

Vetiver grass ( Chrysopogon zizanioides) is a warm season perennial grass grown as a phytoremediation tool and recently proposed as a plant material source for biofuel production. However, limited information exists on Vetiver grass fertility management practices in cropping systems. Therefore, the effects of nitrogen (N) level and N-form on Vetiver growth and development were investigated. Plant slips of C. zizanioides Sunshine were greenhouse-grown using nutrient solution culture. In the first experiment, N-level treatments were 26.3, 52.5, 105, 210 and 410 mg N-L'1 at a ratio of 3:1 nitrate-N: ammo- nium-N. In a second experiment, the ratio of NH4:N03 was changed from 0:100, 25:75, 50:50, 75:25, and 100:0 while keeping a total N level of 210 mg N-L'1. At 12 weeks after transplanting (WAT), plant height, tiller number, accumulated shoot, root, and total fresh weights and dry weights, total leaf number, and chlorophyll content were recorded. Leaf number decreased linearly (P<0.10) from 1 1.2 per plant at 26.3 mg N-L'1 to 9.7 per plant at 210 mg N-L'1 while leaf chlorophyll content increased, then decreased quadratically (P<0.10) over the same N level treatment range. Shoot dry weight and total plant dry weight increased, then decreased (P<0.10, respectively) as N level was increased in nutri¬ ent solution. In the second experiment, plant height decreased linearly (P<0.05) from a mean of 100.9 cm per plant at 1:100 NH4:N03 ratio to 90.5 cm per plant at 100:1 NH4:N03 ratio treatment. Linear increases were observed for tiller number (P<0.10) and leaf number (P<0.001) per plant as the ratio of NH4:NOs was changed from 1:100 to 100:0. Limited effects of form were identified for plant biomass parameters. Results demonstrate that Vetiver grass may successfully be cultured under lower N fertility and has a tolerance to ammonium-N in managed cropping systems.

Key words: chlorophyll, hydroponics, macronutrient, micronutrient, SPAD

INTRODUCTION

Vetiver grass ( Chrysopogon zizanioides L. Roberty) is a perennial aromatic grass native to southern India. Vetiver cultivation is possible in all regions with a warm and moist climate, and it is intensively cultivated in many semitropical areas including China,

2

Indonesia, Angola, Somalia, Congo, Brazil, Guatemala, Haiti, and the southern U.S. (Peyron, 1989). Vetiver grass can tolerate extreme climatic variations such as prolonged drought, flooding and temperatures between -14°C to 55°C. In China, it has survived at -22°C and in the U.S. state of Georgia it was able to tolerate temperatures to -10°C (Troung, 1996). Vetiver is highly tolerant to saline conditions, having a critical salt tox¬ icity level of ECse = 8 dS-m'1 (Troung, 1996). Two genotypes of Vetiver have been identified; namely a seeded north Indian type and a sterile, or very low fertility south Indian type. Sunshine Vetiver is a domesticated South India type which is non-fertile, has received a low risk score for the potential to become invasive by the USDA-Natural Resources Conservation Service, and requires asexual propagation (Joy, 2009). Vetiver grass is currently grown in 100 different countries as a result of World Bank extension efforts (Slinger, 1997).

Vetiver grass has a wide range of ecological and economic applications (Wilde et al., 2005). Applications of Vetiver include use in agriculture land and slope stabilization, soil erosion control, and incorporation in ornamental landscape plantings (Maffei, 2002). It has be inter-cropped as a contour fence with sugar cane ( Saccahrum officinalis), maize (. Zea mays), and tea ( Camellia sinennsis) to conserve soil moisture and restrict soil runoff (Daffron, 1993). Recently Vetiver grass has been proposed as a plant material source for biofuel production (Boucard, 2005). Vetiver grass holds annual biomass production potential of 100 to 120 tonnes-ha'1 (Troung and Smeal, 2003) and has an energy value of 16.3 MJ-kg'1 compared to petroleum (41.9 MJ-kg'1), coal (27.9-30.2 MJ-kg'1), dry wood (19.8 MJ-kg'1), and sugarcane bagasse (9.3 MJ-kg'1; Grimshaw, 2008). It also has poten¬ tial as a carbon sequester and Grimshaw (2008) reported that Vetiver grass can sequester 4.5 times more carbon per year than a fast growing poplar tree ( Populus sp.) per unit area.

The role of using green plants is a promising biological technique for phytoremediation (Schroder et al., 2002). Vetiver has unique morphological and physiological characteris¬ tics that make it an excellent phytoremediation tool (Troung, 2000). In fact, Vetiver has proved to be a more successful phytoremediation plant species than Bermudagrass {Cynodon dactylon ), Bahiagrass ( Paspalum notatum), Rhodes grass ( Chloris guyana), tall wheatgrass ( Thynopyron elongatum), marine couch ( Sporobolus virginicus) and samphire ( Sarcocrina spp.; Sinha et al., 2007).

Nitrogen (N) is an essential element and has a major influence on a number of plant responses including pigmentation, shoot and root growth, cold and drought tolerance, wear tolerance, thatch accumulation and recovery potential (Carrow et al., 1987). An awareness of differences in response to N between economically important grass species underlies much of the present day grassland management and experiments suggest that differences between species in response to N may be considerable (Lovvorn, 1945). Tren- holm et al. (1998) reported that N also affects the development of tillers in grasses. Over fertilization of N can cause grasses to become susceptible to environmental and biologi¬ cal stresses like summer drought, winter desiccation, and heat and cold damage (Dunn et al., 1995). Although most managed crops have recommended fertilizer rates for targeted optimal yield and performance, information on N rate application for Vetiver grass is limited (Joy, 2009). Therefore, this current study was undertaken to investigate the opti¬ mal N fertilization rate and form for improved growth of Vetiver grass.

3

MATERIALS AND METHODS

Experimental Design

Two experiments were carried out in a controlled greenhouse environment (22°C day/14°C night set points) under natural lighting conditions at Illinois State University, Normal, IL (Lat. 40° 30’N) between February and May, 2009. The first study investi¬ gated the effect of nitrogen (N) fertilization level and the second study investigated the effect of N fertilization form on Vetiver grass growth and quality. The experimental design was a randomized complete block design with four replications for each experi¬ ment. Generally, Vetiver grass propagules are available in bundles of rooted slips bearing 3-5 tillers each. For the two experiments, approximately 200 mature plant slips of Sun¬ shine Vetiver grass bearing 3-5 tillers each were purchased from Florida Vetiver Systems LLC (Maitland, FL). Mature plant slip shoots were cut back to 30 cm and roots were trimmed to 20 cm to make them uniform in size at the time of planting. Plastic 38 L containers (Sterilite® Lapis Blue; Townsend, MA) with lids having five 4 cm diameter holes equally spaced 16 cm apart were used for growing plant slips. Each container was filled with 34 L of a modified half strength nutrient solution (Hoagland and Arnon, 1950).

In the N fertilization level study, plants were grown at increasing N treatment concentra¬ tions of 26.3, 52.5, 105, 210, 420 mg NT'1. The two dominant N forms were balanced in all of the N treatments to achieve a ratio of 25% NFl4+:75% N03 . In the N fertilization form study, the total N level was held constant at 210 mg N L"1 and plants were grown under increasing NH4+ at 0, 25, 50, 75 and 100% of total N. Elemental concentrations of nutrient solutions were (mg-L1): phosphorus (P) (89.9); potassium (K) (132); calcium (Ca) (80); magnesium (Mg) (24); iron (Fe) (0.5); boron (B) (0.125); molybdenum (Mo) (0.0025); manganese (Mn) (0.125); and zinc (Zn) (0.0125). Solutions were aerated with an air blower (Model VB-007S, Sweetwater, Ft. Collins, CO) connected to two air stones in each container. Nutrient solutions were replaced every two weeks throughout the experiment to refresh the solution to the initial nutrient concentrations.

Data Collection & Statistical Analysis

After twelve weeks of hydroponic culture, plant height, number of tillers per plant, total number of leaves per plant, and treatment-wise average chlorophyll content of mature leaves using a Minolta SPAD-502 Chlorophyll Meter (Model 2900; Konica Minolta, Japan) were recorded. Plants were then harvested and shoot and root tissues were washed, towel dried, and separated and fresh weights were recorded. Plant tissues were then dried in a forced-air oven at 60°C. When the tissues achieved a constant mass, dry weights were recorded. All data collected were analyzed by the GLM procedure of SAS 9.2 (Cary, NC) to perform analysis of variance and regression analysis to determine relationships between dependent variables and N treatments.

RESULTS AND DISCUSSION

Experiment 1: Nitrogen Fertilization Level Study

Plant height of Vetiver was within previously reported ranges (Xia and Bing, 2000). Increasing N fertilization level influenced Vetiver plant height (P=0.029). Plant height reached a maximum of 105.0 cm after 12 weeks after transplanting under the 210 mg

4

N-L'1 treatment (Table 1). After 14 months of growth, Xia and Bing (2003) reported ‘Sunshine’ Vetiver plant height ranged from 160 cm to 170 cm. However, in the current study, no significant trends were observed for plant height in response to increasing N treatments from 26.3 to 420 mg N-L'1 (Table 1). Non-significant plant height over time of Vetiver was also reported by research performed by Muensangk (2000) which investi¬ gated ecotype differences. Our results indicate that Vetiver can be cultured under N fertil¬ ity as low as 26.3 mg N-L'1 and still attain previously reported average plant height.

The number of tillers per plant was also within previously reported ranges. Increasing N fertilization level influenced the number of tillers per Vetiver plant (P<0.001). Twelve weeks after transplanting, the maximum tiller number was found to be 11.2 at 26.3 mg NL-1 (Table 1). Xia and Bing (2003) reported tiller number per plant ranged from 7 to 59 for twelve different Vetiver ecotypes. Although no significant trends were observed in our current study, the number of tillers decreased from 1 1 .2 at 26.3 mg N-L'1 to 9.7 at 420 mg N-L'1. Abbaszadeh et al. (2009) reported that N level also influenced tiller number in balm ( Mellissa officinalis) and that tiller number per plant decreased as the N level increased.

Increasing N fertilization level influenced Vetiver leaf number per plant (P=0.10) and chlorophyll leaf content (P=0.10). Average leaf number per plant decreased linearly (P<0.10) from 56.0 under 26.3 mg N-L"1 to 46.9 under 420 mg N-L'1 (Table 1). Chloro¬ phyll content displayed a quadratic trend (P<0.10), first increasing then decreasing with increasing N rate concentrations and showed a maximum of 52.7 g-m'2 at 105 mg N-L"1 (Table 1). Buah and Mwinkaara (2009) reported a linear increase in chlorophyll SPAD value content at increasing N levels from 0, 40, 80 and 120 kg N- ha-1 in sorghum ( Sor¬ ghum sp.). Differences in regression trends between Buah and Mwinkaara (2009) and our current study may be attributed to differences in plant genus and level of N fertilization.

Increasing N fertilization level influenced shoot fresh weight (SFWT; P=0.099), shoot dry weight (SDWT; P=0.077), root dry weight (RDWT; P=0.084), and total plant dry weight (TDWT; P=0.096), but not root fresh weight (RFWT; P=0.205) and total plant fresh weight (TFWT; P=0.228). Shoot fresh weight and SDWT were maximum at 52.5 mg N-L'1 (Table 2). Bradshaw et al. (1964) showed that among seven grasses, yields were significantly varied as the N level concentration increased and matgrass (Nardus stricta) produced maximum yield at only 27 ppm (mg-L1) N. Vetiver appears to be another grass species which produces maximum yield under lower total N fertilization. However, Veti¬ ver root dry weight and TDWT were maximum at 210 mg N-L'1. Shoot dry weight increased, then decreased quadratically (P<0.10) in response to increasing N levels (Table 2). Total plant dry weight also increased, then decreased quadratically (P<0.10) in response to increasing N levels (Table 2). Muir et al. (2001) reported that total plant bio¬ mass of switchgrass ( Panicum virgatum ) increased, then decreased quadratically as total N fertilization was increased from 0 kg N-ha'1 to 224 kg N-ha'1.

Experiment 2: Nitrogen Fertilization Form Study

Plant height of Vetiver in this experiment was also within previously reported ranges (Xia and Bing, 2000) and similar to experiment 1 (Table 1). Changing the ratio of NH4+:N03' influenced Vetiver plant height (P=0.001). Plant height reached a maximum of 103.4 cm 12 weeks after transplanting under the 50:50 NH4+:N03 (Table 3). Plant

5

height decreased linearly (P<0.05) as the ratio of NH4+:N03 was changed from 0:100 to 100:0 (Table 3). Increasing ammonium-N fertilization has been reported to decrease plant height in many different plant species (Brown et al., 2010; Muniz et al., 2009; Kim et al., 2006). This is because carbon used for plant growth is reallocated and utilized in the detoxification of absorbed ammonium (Mills and Jones, Jr., 1996).

The number of tillers per plant was also within previously reported ranges (Xia and Bing, 2000) and similar to experiment 1 (Table 1). Changing the ratio of NH4+:N03 influenced Vetiver tiller number (P=0.001) and tiller number increased linearly (P<0.10) from 8.2 tiller per plant at 0% NH4+ to 10.4 tillers per plant at 100% NH4+ 12 WAT (Table 3). Nitrogen form has been demonstrated to affect tillering in grasses (Assuero and Tognetti, 2010), however maximum tillering has been reported in grasses fertilized with a 50:50 NH4+:N03 . Leaf number per plant also increased linearly (P<0.01) in response to increas¬ ing NH4+ in solution (Table 3). A similar trend was reported by Mclnenly et al. (2010) who showed that aboveground biomass of rough fescue ( Festuca campestris ) was increased when foliar N was supplied as 100% ammonium as opposed to 100% nitrate. Cholorphyll leaf content, as measured by SPAD, did not significantly change in response to changing the ratio of NH4+:N03' in solution culture (Table 3).

Changing the ratio of NH4+:N03 influenced RFWT (P=0.10) but did not influenced SFWT (P=0.537), TFWT (P=0.359), SDWT (P=0.566), RDWT (P=0.184), or TDWT (P=0.636). Twelve weeks after transplanting, RFWT decreased linearly (P<0.10) in response to increasing NH4+ in solution (Table 4). When N was supplied as NH4+, bio¬ mass allocation to roots decreased in thickspike wheatgrass ( Agropyron dasystachyum\ Li and Redmann, 1992). Similar results were also reported in creeping bentgrass ( Agrostis palustris ) where root weight was found to be highest at a 1:3 ratio of NH4+:N03' (Glinski et al., 1990). Total fresh shoot and root yields of napiergrass ( Pennisetum purpureum), a tropical grassland species, did not differ when N fertilization treatment were changed from solely nitrate to solely ammonium (Rahman et al., 2010). Bowler and Press (1996) reported that total dry weight of colonial bentgrass ( Agrostis capillaris ) was not affected by N form. The response of Vetiver grass growth to the ratio of NH4+:N03" from 0:100 to 100:0 follows reported trends in other grass species.

Vetiver grass growth and biomass parameters showed varied responses and were mostly non-significant in both the N fertilization level and N fertilization form experiments. This lack of significance may indicate that Sunshine Vetiver has a higher amount of genetic variability than its cultivar classification. According to Grimshaw (2009), Sunshine is a Vetiver cultivar but is also referred to as an upland ecotype. Igbokwe et al. (1991) reported the extent of the fertilizer response among four Vetiver accessions depended on factors such as seasonal variation and genetic variability and Muensangk (2000) reported non-significant trends over time for several growth factors among Vetiver ecotypes. A similar situation has been identified in switchgrass, where two main ecotypes differ in ploidy, morphology, growth pattern, and climatic adaptation (Zalapa et al., 201 1). Adams and Daffron (1997) reported that Monto Vetiver from Australia and Sunshine Vetiver from North America have no morphological and physiological differences and Joy (2009) lists Sunshine as genetically similar to the Vetiver cultivars Boucard, Fort Polk, Haiti, Huffman, Monto, and Vallonia. An ecotypic Sunshine Vetiver population may not respond uniformly to N fertility treatments due to genetic variations. Therefore, further

6

refinement of the current Sunshine population and its related cultivars may be needed prior to large scale production consideration. The same recommendations are being pro¬ posed for switchgrass in order to produce significant heterotic increases in biomass yield in managed cropping systems (Zalapa et al., 201 1).

CONCLUSION

Due to its unique physical and morphological characteristics, its tolerance to environmen¬ tal extremes and low risk of becoming an invasive plant species, Vetiver grass may have production potential in the U.S. as a plant material source for biofuel production or car¬ bon sequestration. Plant height and total plant fresh weight were not significantly differ¬ ent between 26.3 and 420 mg-L'1 N treatments. Therefore, our results indicate that Veti¬ ver grass may successfully be cultured under lower N fertility. Vetiver also showed toler¬ ance to NH4+-N. Although plant height decreased as NH/-N increased, results were still within reported ranges for Vetiver cultivation. No significant differences between 0% and 100% NH4+-N, at a luxuriant total N level, for total plant fresh weight were observed. These results can be used to better predict the optimal N fertilization rates for investigat¬ ing Vetiver grass culture under managed field conditions for large scale production. However, despite the wide range of environmental adaptation and non-fertile reproduc¬ tive characteristics of Sunshine Vetiver, caution should be exercised when introducing a non-native plant species over a wide geographic area. Additional research is also needed to confirm if Sunshine Vetiver grass is actually an ecotype which warrants further genetic refinement through breeding to produce the uniformity of a true cultivar.

ACKNOWLDEGMENTS

This project was funded by support from the Department of Agriculture and College of Applied Science and Technology at Illinois State University. It partially fulfilled the requirement for an M.S. degree for Suneetha Mondyagu completed in May 2010.

LITERATURE CITED

Abbaszadeh, B., H.A. Farahani., S.A. Valadabadi and H.H. Darvishi. 2009. Nitrogenous fertilizer influence on quantity and quality values of balm ( Mellissa officinalis L.). Journal of Agricultural Extension and Rural Development 1 ( 1 ):3 1 -33 .

Adams, R.P. and M.R. Daffron. 1997. DNA fingerprints (RAPDs) of the pan-tropical grass, Vetiveria zizanioides L. The Vetiver Newsletter 18:28-33.

Assuero, S.G. and J.A. Tognetti. 2010. Tillering regulation by endogenous and environmental fac¬ tors and its agricultural management. The Americas Journal of Plant Science and Biotechnology 4(l):35-48.

Boucard, G. 2005. Vetiver Network Discussion Board, (http://www.vetiver.org/cgi-bin/discus /discus. cgi).

Bowler, J.M. and M.C. Press. 1996. Effects of elevated C02, nitrogen form and concentration on growth and photosynthesis of a fast- and slow-growing grass. New Phytologist 1 32(3):39 1 -40 1 . Bradshaw, A.D., M.J. Chadwick, D. Jowett and R.W. Snaydon. 1964. Experimental investigations in to the mineral nutrition of several grass species. JSTOR: Journal of Ecology 52(3):665-676. Brown, H.E., M.S. Watt, P.W. Clinton, and E.G. Mason. 2010. Influence of ammonium and nitrate supply on growth, dry matter partitioning, N uptake and photosynthetic capacity of Pinus radiata seedlings. Trees 24(6): 1097-1 107.

Buah, S.S.J. and S. Mwinkaara. 2009. Response of sorghum to nitrogen fertilizer and plant density in the guinea savanna zone. Journal of Agronomy 8(4): 124-130.

7

Carrow, R.N., B.J. Johnson and R.E. Burns. 1987. Thatch and quality of Tifway bermudagrass in relation to fertility and cultivation. Agronomy Journal 79:524-530.

Daffron, M. 1993. Question and Answers, p. 59-70. In: Vetiver grass: A thin green line against erosion. National Academy Press, Washington, D.C.

Dunn, J.H., D.D. Minner, B.F. Fresenburg, S.S. Bughrara, and C.H. Hohnstrater. 1995. Influence of core aerification, topdressing, and nitrogen on mat, roots, and quality of Meyer zoysiagrass. Agronomy Journal 87(5):89 1-894.

Glinski, D.S., H.A. Mills, K.J. Karnok and R.N. Carrow. 1990. Nitrogen form influences root growth of sodded creeping bentgrass. HortScience 25(8):932-933.

Grimshaw, R. 2008. First National Indian Vetiver Workshop. The Vetiver Network International. (www.vetiver.org).

Grimshaw, R. 2009. Personal communication, (http://www.vetiver.org/cgi-bin/discus/discus.cgi).

Hoagland, D.R. and D.J. Arnon. 1950. The water culture method for growing plants without soil. California Agricultural Experimental Station Circular 347. Berkeley, CA.

Igbokwe. P.E., S.C. Tiwari, J.L. Burton and R.E. Waters, Jr. 1991. Influence of accession variabil¬ ity and fertilization on the establishment and growth of Vetiver grass in a field nursery. The Vetiver Network Newsletter 7:113-125.

Joy, R.J. 2009. ‘Sunshine’ vetivergrass Chrysopogon zizanioides (L.) Roberty. USDA-NRCS Pacific Island Area Plant Materials Program, Hoolehuam HI.

Kim, S.J., C. Kawahaarada, and G. Ishii. 2006. Effect of ammonion:nitrate nutrient ratio on nitrate and glucosinolate contents of hydroponically-grown rocket salad ( Eruca sativ a Mill.). Soils Sci¬ ence and Plant Nutrition 52(3):387-393.

Li, Y.S. and R.E. Redmann. 1992. Growth-responses to ammonion-N, nitrate-N and defoliation in Agropyron dasystachyum from the Canadian mixed grassland. American Midland Naturalist 127(2):300-308.

Lovvom, R.L. 1945. The effect of defoliation, soil fertility, temperature and length of day on growth of some perennial grasses. Journal of American Society Agronomy 37:570-582.

Maffei, M. 2002. Introduction to genus Vetiveria, p. 1-18. In: M. Maffei (ed.). Vetiveria: The genus Vetiveria. Taylor & Francis, London, UK.

Mclnenly, L.E., E.H. Merrill, J.F. Cahill, and N.G. Juma. 2010. Festuca campestris alters root morphology and growth in response to simulated grazing and nitrogen form. Functional Ecology 24(2):283-292.

Mills, H.A. and J.B. Jones, Jr. 1996. Essential plant nutrients, p. 6-62. In: Plant analysis handbook II. MicroMacro Publishing Inc., Athens, Georgia, USA.

Muensangk, S. 2000. The use of Vetiver grass in the reclamation and improvement of acid sulphate soils in Thailand: A pre-experiment study. In: Proceedings of the second international Vetiver conference, Thailand. Part 4:279-282.

Muir, P.J., M.A. Sanderson, W.R. Ocumpaugh, R.M. Jones and R.L. Reed. 2001. Biomass produc¬ tion of ‘Alamo’ switchgrass in response to nitrogen, phosphorus and row spacing. Agronomy Journal 93:896-90.

Muniz, M.A., J.G. Barbosa, J.A.S. Grossi, M.Y. Orbes, and P.G. Sa. 2009. Production and quality of pot chrysanthemum fertigated with different nitrate/ammonium relations. Bioscience Journal 25(l):75-82.

Peyron, L. 1989. Vetiver in perfumery. Quintessenza 13:4-14.

Rahman, M.M., Y. Ishii, M. Niimi, and O. Kawamura. 2010. Effect of application form of nitrogen on oxalate accumulation and mineral uptake by napiergrass ( Pennisetum purpureum). Grassland Science 56(3): 141 - 144.

Schroder, P., P.J. Harvey and J.P. Schwitzguebel. 2002. Prospects for the phytoremediation of organic pollutants in Europe. Environmental Science Pollution R 9(1): 1-3.

Sinha, R.K., H. Sunil, and P.K. Tandon. 2007. Phytoremediation: Role of plants in contaminated site management, p. 315-330. In: S.N. Singh and R.D. Tripathi (eds.). Environmental bioremediation technologies. Springer- Verlag, New York, NY.

Slinger, V. 1997. Introduction to genus Vetiveria, p. 1-18. In: M. Maffei (ed.). Vetiveria: The genus Vetiveria. Taylor & Francis, London, UK.

Trenholm, L.E., A.E. Dudeck, J.B. Sartian, and J.L. Cisar. 1998. Bermudagrass growth, total nonstructural carbohydrate concentration, and quality as influenced by nitrogen and potassium. Crop Science 38:168-173.

8

Troung, P.N. 1996. An overview of research, development and application of the VGS overseas and in Queensland, p.6-7. In: Proc. R, D and A of VGS in Queensland, Australia.

Troung, P.N. 2000. The global impact of Vetiver grass technology on the environment, p. 46-47. In: Proceedings of the second international Vetiver conference, Thailand.

Troung, P.N. and C. Smeal. 2003. Pacific Rim Vetiver Network Tech. Bull. No. 2003/3.

Wilde, E.W., R.L. Brigmon, D.L. Dunn, M.A. Heitkamp, and D.C. Dagnan. 2005. Phytoextraction of lead from firing range soil by Vetiver grass. Chemosphere 61:1451-1457.

Xia, H.P. and Y. B. Bing. 2003. Study of screening better ecotypes of Vetiver grass, p. 517-523. In: Proceedings of third international conference on Vetiver and exhibition. China Agriculture Press, Guangzhou, China.

Zalapa, J.E., D.L. Price, S.M. Kaeppleer, C.M. Tobias, M. Okada, and M.D. Casler. 2011. Hierar¬ chical classification of switchgrass genotypes using SSR and chloroplast sequences: Ecoytpes, ploides, gene pools, and cultivars. Theoretical and Applied Genetics 122:805-817.

9

Table 1. Mean values2 of Vetiver grass plant height, tiller number, leaf number, and chlorophyll content 12 weeks after transplanting (WAT) into a modified Hoa- gland’s nutrient solution with nitrogen levels (N LEVEL) varying from 26.3 mg-L'1 to 420 mg-L'1 with ratio of 1:3 NH4:N03.

N LEVEL (mg-L1)	Plant heighty (cm)	Tiller Number	Leaf Number	ChlorophylP (mg-m'2)
26.3	100.1 ± 17.6	11.2 ±4.8	56.00 ±6.13	48.42 ±3.42
52.5	100.4 ± 13.4	10.5 ±4.9	49.25 ±2.93	48.62 ± 1.37
105	102.4 ± 13.0	10.7 ±5.7	53.05 ±11.68	52.72 ± 1.69
210	105.0 ±8.7	10.4 ±3.3	47.25 ±5.20	48.52 ±2.89
420 Contrast	99.8 ± 12.2	9.7 ±3.9	46.95 ± 10.35	48.37 ±2.85
Linear	NS	NS	*	NS
Quadratic	NS	NS	NS	*

2 Mean of 5 plants per treatment replication ± standard deviation. y Plant height was measured from base of the crown to leaf tip in centimeters (cm). xMean chlorophyll content of mature leaves measured in mg-m'2 using a hand-held SPAD meter.

^Significance for linear and quadratic orthogonal contrasts for N LEVELS. ns,*,**,*** Non-significant or significance at P<0.10, P<0.05, P<0.01, respectively.

Table 2. Mean values2 of Vetiver grass plant biomass 12 weeks after transplanting into a modified Hoagland’s nutrient solution with nitrogen levels (N LEVEL) varying

	from 26.3 mg-L	1 to 420 mg-L	1 with a ratio of 1:3 NH4:N03.
N LEVEL			Plant biomassy (g)
(mg-L'1)	SFWT	RFWT	TFWT	SDWT	RDWT	TDWT
26.3	387.5 ±75.0	233.9 ±50.1	621.4 ± 119.9	77.2 ± 14.0	35.3 ±5.5	112.5 ± 18.5
52.5	406.4 ± 62.0	215.8 ± 14.2	622.1 ± 101.2	85.2 ± 14.9	34.7 ± 2.3	1 19.9 ± 16.6
105	391.3 ±73.5	264.1 ±69.6	655.5 ± 141.5	82.1 ± 12.9	37.2 ±6.3	119.3 ± 18.9
210	388.7 ±21.4	263.8 ±24.7	652.5 ± 19.6	82.9 ±5.3	37.7 ± 1.9	120.6 ±5.4
420	345.7 ± 12.4	246.9 ±27.7	592.7 ± 33.9	71.5 ±2.8	34.1 ±2.0	105.6 ±3.8
Contrast14
Linear	NS	NS	NS	NS	NS	NS
Quadratic	NS	NS	NS	*	NS	*

2 Mean of 5 plants per treatment replication ± standard deviation. Abbreviations: SFWT=shoot fresh weight, RFWT=root fresh weight, TFWT=total plant fresh weight, SDWT=shoot dry weight, RDWT=root dry weight, and TDWT=total plant dry weight. y Mean plant biomass measured in grams (g).

x Significance for linear and quadratic orthogonal contrasts for N LEVELS. ns,*,**,*** Non-significant or significance at P<0.10, P<0.05, P<0.01 , respectively.

10

Table 3. Mean values2 of Vetiver grass plant height, tiller number, leaf number, and chlorophyll content 12 weeks after transplanting (WAT) into a modified Hoa- gland’s nutrient solution with varying NH4:N03 ratios of 0:100, 25:75, 50:50, 75:25, 100:0 while keeping total N level constant at 210 mg N-L'1.

RATIO of NH4:N03	Plant heighty (cm)	Tiller number	Leaf Number	Chlorophyll" (mg-m 2)
0:100	100.9 ± 14.1	8.2 ±3.2	36.2 ±2.6	45.4 ±3.6
25:75	102.5 ± 11.3	9.2 ±4.2	40.6 ±5.1	46.0 ±2.9
50:50	103.4 ±9.8	9.1 ±3.8	42.1 ±8.2	48.9 ±4.2
75:25	100.6 ±9.3	9.5 ±3.7	42.3 ±3.1	48.8 ±3.3
100:0 Contrast™	90.5 ± 14.5	10.4 ±4.2	44.9 ±3.3	47.1 ±4.3
Linear	**	*	***	NS
Quadratic	***	NS	**	NS

zMean of 5 plants per treatment replication ± standard deviation. y Plant height was measured from base of the crown to leaf tip in centimeters (cm). xMean chlorophyll content of mature leaves measured in mg-m'2 using a hand-held SPAD meter.

Significance for linear and quadratic orthogonal contrasts for N RATIOS. ns,*,**,*** Non_sigmficant or significance at P<0.10, P<0.05, P<0.01, respectively.

Table 4. Mean values2 of Vetiver grass plant biomass 12 weeks after transplanting into a modified Hoagland’s nutrient solution with varying NH4:N03 ratios of 0:100, 25:75, 50:50, 75:25, 100:0 while keeping total N rate constant at 210 mg NT1.

RATIO of Plant biomassy (g)

NH4:N03	SFWT	RFWT	TFWT	SDWT	RDWT	TDWT
0:100	357.3 ±45.2	208.5 ±25.5	565.7 ±69.3	75.9 ±6.7	32.5 ±0.9	108.3 ±7.6
25:75	383.9 ±53.6	236.2 ± 66.8	620.2 ±109.8	84.9 ± 13.5	34.2 ±5.9	119.1 ± 17.6
50:50	418.7 ±68.6	204.1 ±47.8	622.8 ± 112.2	88.3 ± 11.9	31.6 ± 6.1	1 19.9 ± 17.1
75:25	351.5 ±35.1	202.2 ±73.8	553.7 ± 108.7	75.9 ±5.9	31.1 ±6.7	107.1 ± 10.0
100:0	349.8 ±43.4	161.3 ±9.2	511.1 ±45.6	82.6 ± 13.4	29.2 ± 1.0	1 1 1 .8 ± 12.9
Contrast11
Linear	NS	*	NS	NS	NS	NS
Quadratic	NS	NS	NS	NS	NS	NS
2 Mean of 5 plants per treatment	replication ±	standard deviation. Abbreviations

SFWT=shoot fresh weight, RFWT=root fresh weight, TFWT=total plant fresh weight, SDWT=shoot dry weight, RDWT=root dry weight, and TDWT=total plant dry weight. y Mean plant biomass measured in grams (g).

x Significance for linear and quadratic orthogonal contrasts for N RATIOS.

VJQ jjc £££

’ ’ ’ Non-significant or significance at P<0.10, P<0.05, P<0.01 , respectively.

Transactions of the Illinois State Academy of Science (2012) Volume 105,#l&2,pp. 11-18

received 12/6/1 1 accepted 3/29/12

Flora of Twin Shelters and Twin Mounds Hill Prairies, Pere Marquette State Park,

Jersey County, Illinois, Changes Since 1963

William E. McClain1, John E. Ebinger2

'Illinois State Museum, Springfield, Illinois 62706; department of Biological Sciences,

Eastern Illinois University, Charleston, Illinois 61920

ABSTRACT

The vascular flora of Twin Mounds and Twin Shelters hill prairies within Pere Marquette State Park, Jersey County, Illinois was studied during the 2009 and 2010 growing sea¬ sons. These two prairies are located on southwest-facing slopes approximately 10 km from Grafton, Illinois. Community composition was analyzed using m2 quadrats placed at one-meter intervals along two randomly located transect lines within each prairie. Fre¬ quency, mean cover, and importance value (I. V. total = 200) were determined from the data collected. A total of 59 vascular plant taxa was collected in the prairies with 32 encountered in plots. Schizachyrium scoparium (Michx.) Nash (little bluestem) had the highest importance value followed by Aster oolentangiensis (sky blue aster), Sor- ghastrum nutans (L.) Nash (Indian grass), and Solidago nemoralis Ait. (gray goldenrod). Changes in the vegetation within the last 47 years include the extirpation of two vascular plant species, Lespedeza violacea (L.) Pers. (violet lespedeza) and Asclepias hirtella (Pennell) Woodson (tall green milkweed) and the moss Wiessia controversa Hedwig. The lichen Dermatocarpon hepaticum (Ach.) Th. Fr. and the vascular plant Senecio plattensis Nutt, (prairie ragwort) experienced significant population declines while the combined biomass of Andropogon gerardii Vitman (big bluestem) and Sorghastrum nutans (L.) Nash (Indian grass) increased.

INTRODUCTION

Pere Marquette State Park lies within the Driftless Section of the Middle Mississippi Bor¬ der Natural Division (T6N, R13W, Sec. 9). This division is characterized by steep topography and numerous outcrops of limestone, dolomite, and shale, especially within Pere Marquette State Park (Schwegman 1973). The most widespread geologic feature is Pleistocene loess which forms a thick mantle over the underlying bedrock (Rubey 1952). This loess is the parent material for Hamburg silt, the grass-covered, calcareous soil characteristic of hill prairie communities in the western portion of Illinois. Though most of Pere Marquette State Park is forested, hill prairie communities are conspicuous fea¬ tures of south- to southwest-facing bluffs bordering the Illinois River floodplain (McFall & Karnes 1995). These hill prairies have been the focus of several studies, including Kil-

12

burn and Ford (1963) who studied the flora of Twin Mounds hill prairie, McClain (1983) who documented the loss of hill prairie at five locations in Pere Marquette State Park, and McClain and Anderson (1990) who studied woody invasion on Twin Mounds hill prairie. In addition, Evers (1955) examined numerous hill prairies in the Mississippi and Illinois river systems. More than 50 hill prairies throughout Illinois were examined in his exten¬ sive study and dominant plant species, disturbances, and general characteristics were rec¬ orded.

These previous studies, like most hill prairie surveys, were not designed to detect long term trends in the vegetation. The purpose of the present study was to document the flora of Twin Mounds and Twin Shelters hill prairies, and to determine changes in the vegeta¬ tion of Twin Mounds hill prairie since the study of Kilburn and Ford (1963).

DESCRIPTION OF THE STUDY SITE

Land purchases for the establishment of Pere Marquette State Park began in 1932. Build¬ ing and trail construction, including the shelter at the crest of Twin Shelters Hill Prairie, were completed by the Civilian Conservation Corps (CCC) under the direction of the National Park Service during the period April 1, 1933 to June 30, 1939. The work of the CCC also included 96 days for fighting “weed fires”, the construction of four miles of firebreaks, 11 miles of fire hazard reduction, and 286 days of fire suppression training (McClain and Anderson 1990).

Fire suppression was practiced at Pere Marquette State Park and none of the hill prairies are known to have burned during the forty year period from 1932 until 1972. Woody vegetation made considerable advances onto the hill prairies during this time. Twin Mounds and Twin Shelters hill prairies are currently about 100 m apart, but were part of one large prairie in the 1930s based on 1936 aerial photographs (McClain 1983). Woody invasion reduced the size of this large prairie from 8.7 ha in 1936 to 1.5 ha in 1983 (McClain and Anderson 1990). A “thin thread” of prairie still connected these two sites in 1979, but this narrow remnant has since been obscured by woody vegetation (McClain, personal observations). Recent studies indicate a reduction of 50% to 65% in the size of many hill prairies during the 50 year period from 1936 to 1986 (McClain and Anderson 1990, Robertson et. al. 1996, Schwartz et. al. 1997).

Prescribed fire was introduced to Pere Marquette in 1973 when a burn was conducted on Twin Mounds hill prairie by the Illinois Department of Conservation (now the Depart¬ ment of Natural Resources). Subsequent prescribed burns were conducted on the Twin Mounds hill prairie in 1975, 1977, 1982, 1985, 1987, 1996, 1999, 2002, 2007, 2009, and 2011. Prescribed burns were conducted on Twin Shelters hill prairie in 1974, 1987, 1996, 2007, and 2009.

METHODS

Twin Mounds and Twin Shelters hill prairies were visited during the growing seasons of 2009 and 2010 to collect vascular plant specimens and study the composition and struc¬ ture of the prairie vegetation. Voucher specimens were deposited in the Stover-Ebinger Herbarium of Eastern Illinois University in Charleston (EIU). Exotic species were identi-

13

fied using Gleason and Cronquist (1991) and Mohlenbrock (2002) while nomenclature follows Mohlenbrock (2002).

The ground layer vegetation was surveyed along two 25 m long transects (n = 50 at each site) located on the mid-slope of Twin Mounds and Twin Shelters hill prairies. The tran¬ sect location at Twin Mounds hill prairie is within the area studied by Kilburn and Ford (1963). One m2 quadrats were located along each transect at 1 m intervals (n = 25). Odd- numbered quadrats were placed to the right and even-numbered quadrats were placed to the left. A random numbers table was used to determine the number of meters (0-9) a quadrat was placed from transects. Cover was determined using the Daubenmire cover class system (Daubenmire 1959) as modified by Bailey and Poulton (1968). From these data, frequency (%), relative frequency, mean cover (% of total cover), relative cover, and Importance Values (relative frequency + relative cover) were determined for each species found in the plots.

RESULTS

A total of 59 vascular plant species representing 25 families and 49 genera were docu¬ mented on the prairies, including 1 1 monocot species representing 3 families and 48 dicot species representing 22 families (Appendix 1). The most common families were the Asteraceae with 13 species, Poaceae (9), and Fabaceae (9). No threatened or endangered species was discovered during the survey, while five exotic species were encountered.

Schizachyrium scoparium (Michx.) Nash, (little bluestem) dominated both prairies, accounting for 38.5 percent of the importance value at Twin Mounds hill prairie and 44.6 percent at Twin Shelters hill prairie (Table 1). Sorgastrum nutans (L.) Nash (Indian grass) was third in importance at Twin Mounds hill prairie and second at Twin Shelters hill prairie. Other common native grasses included Bouteloua curtipendula (Michx.) Torr. (side-oats grama) and Andropogon gerardii Vitman (big bluestem), both species with importance values of 8 or more. Aster oolantangiensis Riddell (sky-blue aster) was the most important forb encountered, being second in importance value (36.6) at Twin Mounds hill prairie and third in importance value (28.3) at Twin Shelters hill prairie. Solidago nemoralis Ait. (gray goldenrod), Desmodium sessilifolium (Torr.) Torr. & Gray (sessile-leaved tick trefoil), and Dalea purpurea Vent, (purple prairie clover) were other common forbs. Only 26 species, with a mean total cover of 60.4%, were recorded for the plots on Twin Mounds hill prairie, while 25 species, with a mean total cover of 54.9%, were reported on Twin Shelters hill prairie. Bare ground and litter cover values were nearly identical at both sites, varying from 41% to 43 % (Table 1). Small numbers of an unidentifiable species of the lichen Dermatocarpon, possibly D. hepaticum (Ach.) Th. Fr., and clumps of the moss Barbula convuluta Hedwig were present near the base of Twin Mounds hill prairie. No moss or lichen species were encountered in study plots on either site.

DISCUSSION

The vegetation of the study sites is not diverse. Only 59 vascular plant species were found during this study (9 trips over a two year period) compared to 65 recorded by Kil¬ burn and Ford (1963). Four vascular plant species listed in 1963, Desmodium panicula-

14

turn (Nutt.) DC (panicled tick trefoil), Spiranthes gracilis (Bigel) Beck (slender ladies tresses) Lespedeza violacea (L.) Pers. (violet bush clover) and Asclepias hirtella (Pen¬ nell) Woodson (tall green milkweed), could not be located in 2010 and appear to be extir¬ pated. Sessile-leaved tick trefoil, a species present on both hill prairies in 2010, may have been confused with panicled tick trefoil. Spiranthes magnicamporum Sheviak (prairie ladies’ tresses), a species found in the present study, was not recognized until 1973 (She¬ viak 1973). It is a fall blooming species characterized by two to three spiraling rows of flowers compared to just one row for slender ladies’ tresses (Mohlenbrock 2002).

The two hill prairies have nearly identical vascular plant species compositions. All but one of the 59 species, including five exotic taxa, recorded for Twin Shelters hill prairie were also present on Twin Mounds hill prairie (Appendix 1). Gray goldenrod, sessile¬ leaved tick trefoil, and purple prairie clover were more common on Twin Mounds hill prairie while Aster oblongifolius Nutt, (aromatic aster), Hedyotis nigricans, and big bluestem were more abundant on Twin Shelters hill prairie. Twin Mounds hill prairie occupies the highest, most exposed part of the bluff and has experienced more manage¬ ment (twelve prescribed burns compared to five), factors that could contribute to the higher wildflower densities at this site (Table 1).

Kilburn and Ford (1963) did not report any Aster species. However, Asters were abun¬ dant in 2010 and they were reported from the early 1950s (Evers 1955). Four Aster spe¬ cies were collected in 2010, including three in study plots. Aster oolantangiensis Riddell (sky blue aster) was most common, having importance values of 36.6% at Twin Mounds hill prairie and 28.3% at Twin Shelters hill prairie (Table 1). The reasons for their absence in the 1963 study are not known, but Aster species may have been overlooked if sampling was conducted when plants were not flowering.

The increase in biomass of the tall prairie grasses, big bluestem and Indian grass, since 1963 may be affecting the abundance of small-statured plants (rosettes of vascular plants, mosses and lichens) that inhabit open, sunny spaces between clumps of prairie grasses. Big bluestem and Indian grass had combined cover values on Twin Mounds hill prairie of only 3.4% in 1963 compared to 14.3 % in 2010 (Table 1). Prairie ragwort, listed as S. pauperacalus Michx. (balsam ragwort) by Kilburn and Ford (1963), was common in quadrats on Twin Mounds hill prairie in 1963 with a frequency of 87 %. Its frequency dropped to 27 % by 1990 (McClain & Anderson 1990), and no plants were present in study plots on either prairie in 2010 (Table 1). Only nine plants were found growing in small patches of open soil near the base of Twin Mounds hill prairie

Nonvascular plants have also been affected by changes in the prairie community. The moss Weisia controversa Hedwig and the lichen Dermatocarpon hepaticum were promi¬ nent in 1963, having frequencies of 87.8 % and 68.9 %. However, neither species was present in plots in 2010 (Kilburn & Ford 1963, Table 1). Weisia controversa Hedwig appears to be extirpated, but a small population of Dermatocarpon, possibly D. hepati¬ cum , was found at the base of Twin Shelters hill prairie in the same eroded area as prairie ragwort.

Prescribed fire programs have been implemented on hill prairies to mitigate the loss of wildland fire. However, most prescribed burns are conducted during the dormant season,

15

a practice that favors grasses such as big bluestem and Indian grass over wildlflowers. The hill prairies at Pere Marquette State Park were among the first sites burned when the Illinois Department of Conservation (Now the Department of Natural Resources) began its prescribed burning program in 1973. The cumulative effects of dormant season burn¬ ing over a period of nearly forty years may be contributing to the prominent increase in biomass of tall prairie grasses (Anderson 2006).

The abundance of whitetail deer may also be affecting herbaceous wildflower density in these two hill prairies. These animals have become abundant in Illinois in the last 50 years and evidence of whitetail deer was observed in both hill prairies during this study. Deer selectively graze wildflowers such as Amorpha canescens Pursh. (lead plant), but rarely graze grasses or other graminoids. Deer browsing, which has the potential to shift herbaceous prairie plant populations in favor of grasses, may be contributing to the prominence of tall prairie grasses (Anderson et. al. 2001).

The hill prairies of Illinois are sites of rapid change. Woody invasion, fragmentation, size reduction, and the complete disappearance of these grasslands have taken place in the last 60 years (Robertson et. al. 1996, Schwartz et. al. 1997, McClain et. al. 2009). This study of hill prairies at Pere Marquette State Park suggests that other vegetation changes are occurring along with fragmentation and woody invasion, including changes in the density of grasses and wildflowers and species loss (McClain et. al. 2004).

LITERATURE CITED

Anderson, R. C., E. A. Corbett, M. R. Anderson, G.A. Corbett, and T. M. Kelley. 2001. High white-tailed deer density has negative impact on prairie forbs. Journal of the Torrey Botanical Club 128(4): 381-392.

Anderson, R. C. 2006. Evolution and origin of the grassland of Central North America: climate, fire, and mammalian grazers. Journal of the Torrey Botanical Society 133(4): 626-647.

Bailey, A. W. and C. E. Poulton. 1968. Plant communities and environmental relationships in a portion of Tillamook burn, northwestern Oregon. Ecology 49: 1-13.

Daubenmire, R. 1959. A canopy coverage method of vegetation analysis. Northwest Science 33: 43-64.

Evers, R. A. 1955. Hill Prairies of Illinois. Illinois Natural History Survey Bulletin 26: 367-446. Gleason, H. A. and A. Cronquist. 1991. Manual of Vascular Plants of Northeastern United States and Adjacent Canada. Second Edition, The New York Botanical Garden, Bronx, New York. Kilburn, P. D. and C. D. Ford, Jr. 1963. Frequency distribution of hill prairie plants. Transactions of the Illinois Academy of Science 56:94-97.

McClain, W. E. 1983. Photodocumentation of the loss of hill prairie within Pere Marquette State Park, Jersey County, Illinois. Transactions of the Illinois State Academy of Science 76:343-346. McClain, W. E. and E. A. Anderson. 1990. Loss of hill prairie through woody plant invasion at Pere Marquette State Park, Jersey County, Illinois. Natural Areas Journal 10:69-75.

McClain, W. E., Moorehouse, A., Ebinger, J. E. 2009. Loss of Hulsebus Hill Prairie, Henderson County, Illinois, to woody plant invasion. Transactions of the Illinois State Academy of Science 102 (3& 4): 141- 147.

McClain, W. E., Phillippe, L. R., Ebinger, J. E. 2004. Vascular Flora of Manito Prairie Nature Pre¬ serve, Tazewell County, Illinois. Transactions of the Illinois State Academy of Science 97(2): 83-94.

McFall, D. and J. Karnes. 1995. (Editors). A directory of Illinois Nature Preserves. Volume 2. Northwestern, Central and Southern Illinois. Illinois Department of Natural Resources, Division of Natural Heritage, Springfield, Illinois.

16

Mohlenbrock, R. H. 2002. Vascular flora of Illinois. Southern Illinois University Press, Carbondale, Illinois.

Robertson, K. R., M. W. Schwartz, J. W. Olson, B. K. Dunphy, and H. D. Clarke. 1996. Fifty years of change in Illinois Hill Prairies. Erigenia 14:41-52.

Rubey, W. W. 1952. Geology and mineral resources of the Hardin and Brussels Quadrangles. U. S. Government Printing Office. Washington, D. C.

Schwartz, M. W., K. R. Robertson, B. K. Dunphy, J. W. Olson, and A. M. Trame. 1997. The biogeography of and habitat loss on hill prairies. Pages 267-283. in M. M. Schwartz, ed., Conservation of Highly Fragmented Landscapes. Chapman & Hall. New York.

Schwegman, J. E. 1973. Comprehensive plan for the Illinois Nature Preserves System, Part 2. The Natural Divisions of Illinois. Illinois Nature Preserves Commission, Rockford. 32 pp + map.

Sheviak, C.J. 1973. A new Spiranthes from the grasslands of Central America. Botanical Museum Leaflet 23: 285-297.

17

Table 1. Frequency (%), mean cover (% of total area), and importance value (IV) for the species encountered at Twin Mounds and at Twin Shelters Loess Hill Prairies, Pere Marquette State Park, Jersey County, Illinois (*exotic species)

Twin Mounds Prairie Twin Shelters Prairie

Species	Freq. %	Mean Cover	I. V.	Freq %	Mean Cover	I.V.
Schizachyrium scoparium	100	16.11	38.5	94	17.43	44.6
Aster oolentangiensis	100	15.00	36.6	84	9.24	28.3
Sorghastrum nutans	92	11.83	30.5	84	10.91	31.4
Soli dago nemoralis	80	6.43	20.2	40	2.78	10.6
Desmodium sessilifolium	74	3.11	13.8	42	0.71	7.0
Dalea purpurea	92	1.64	13.6	44	0.62	7.1
Bouteloua curtipendula	96	1.03	13.1	78	0.74	11.9
Andropogon gerardii	34	2.46	8.1	52	5.40	16.9
Aster ericoides	42	1.53	7.5	12	0.06	1.7
Brickellia eupatorioides	24	0.22	3.2	24	0.27	3.8
Desmanthus illinoensis	16	0.18	2.2	12	0.11	1.8
Ruellia humilis	16	0.13	2.1	14	0.17	2.2
Hedyotis nigricans	14	0.07	1.8	52	2.07	10.9
Desmodium ciliare	12	0.16	1.7	8	0.09	1.3
Agalinus aspera	10	0.05	1.3	4	0.02	0.5
Asclepias viridiflora	6	0.08	0.8	—	—	—
Dalea Candida	6	0.08	0.8	2	0.01	0.3
Dichanthelium	6	0.03	0.8	—	—	—
oligosanthes
Spiranthes	6	0.03	0.8	—	—	—
magnicamporum
Asclepias verticillata	4	0.07	0.6	4	0.02	0.5
Linum sulcatum	4	0.02	0.5	4	0.02	0.5
Lithospermum incisum	4	0.02	0.5	—	—	—
Cornus drummondii	2	0.06	0.3	6	1.11	2.8
Eupatorium altissimum	2	0.06	0.3	—	—	—
Amorpha canescens	2	0.01	0.2	—	—	—
Antennaria plantaginifolia	2	0.01	0.2	—	—	—
Aster oblongifolius	—	—	—	34	2.02	8.3
Lespedeza virginica	—	—	—	10	0.49	2.3
Rhus aromatic	—	—	—	8	0.43	1.9
Carex pensylvanica	—	—	—	10	0.10	1.6
Pycnanthemum pilosum	—	—	—	8	0.09	1.3
Lithospermum canescens	—	—	—	4	0.02	0.5
Totals		60.42	200.0		54.93	200.0
Bare ground and litter		41.24			43.11

18

APPENDIX I

Vascular plant species encountered on two loess hill prairies at the Pere Marquette State Park, Jersey County, Illinois are listed alphabetically by family under the major plant groups. Collecting numbers are preceded by the initial of the collector (E = John E. Ebinger; M = William E. McClain). Specimens are deposited in the Stover/Ebinger Herbarium (EIU), Eastern Illinois University, Charleston, Illinois. (*exotic species)

DICOTS

Acanthaceae

Ruellia humilis Nutt.: E32349; E32356

Anacardiaceae

Rhus aromatica Ait. E32348; E32359

Asclepiadaceae

Asclepias verticillata L.: E32329; E32372 Asclepias viridiflora Raf. E32333; E32368

Asteraceae

Antennaria plantaginifolia (L.) Hook. E32679 Aster ericoides L.: E32675 Aster oblongifolius L.: E32684 Aster oolentangiensis Riddell: E32683 Aster pilosus Willd.: E32676 Brickellia eupatorioides (L.) Shinners: E32345; E32355

Erigeron strigosus Muhl.: E32228; E32222 Eupatorium altissimum L.: E32680 Eupatorium serotinum Michx.: E32353 Helianthus divaricatus L.:E32331 Senecio plattensis Nutt. M2829 Solidago nemoralis Ait.: E32330; E32367 Vernonia missourica Raf.: E32350 Boraginaceae

Lithospermum canescens (Michx.) Lehm.: E32685

Lithospermum incisum Lehm.: E32678 Caeslpiniaceae

Chamaecrista nictitans (L.) Moench.:E32335

Campanulaceae

Lobelia spicata Lam.: E32334: E32376;

M2763

Cornaceae

Cornus drummondii C.A. Mey.: E32342 ; E32375

Euphorbiaceae

Chamaesyce nutans (Lag.) Small: E32363 Croton monanthogynus Michx.: E32361

Fabaceae

Amorpha canescens Pursh: E32336; E32371 Dalea Candida (Michx.) Willd.: E32346 Dalea purpurea Vent.: E32338; E32358 Desmodium ciliare (Muhl.) DC.: E32340; E32362

Desmodium sessilifolium (Torr.) Torr. & Gray: E32339; E32360

Lespedeza capitata Michx.: E32677 Lespedeza virginica (L.) Britt.: E32682 *Melilotus albus Medic.: E32343; E32370

Psoralidium tenuiflorum (Pursh) Rydb.: E32229; E32223

Hypericaceae

*Hypericum perforatum L.: M2762

Lamiaceae

Monarda fistulosa L.: E32351 Pycnanthemum pilosum Nutt.: E32337

Linaceae

Linum sulcatum Riddell.: E32347: E32373

Mimosaceae

Desmanthus illinoensis (Michx.) MacM.: E32369

Onagraceae

Oenothera biennis L.: E32366

Oxalidaceae

Oxalis stricta L.: E32224

Ranunculaceae

Anemone virginiana L.: E32230

Rubiaceae

Hedyotis nigricans (Lam.) Fosb.: E32354; E32357

Scrophulariaceae

Agalinus aspera (Doug.) Britt.: E32687 Penstemon pallidus Small: E32231; E32225

Simaroubaceae

*Ailanthus altissima (Mill.) Swingle: E32344

Solanaceae

Physalis heterophylla Dunal.: E32341; E32364 Physalis virginiana Mill.: E32232 Verbenaceae

Verbena stricta Vent.: M2761

MONOCOTS

Cyperaceae

Carex pensylvanica Lam.: E32686

Orchidaceae

Spiranthes magnicamporum (L.) Rich.: E32688

Poaceae

Andropogon gerardii Vitman: E32712 Bouteloua curtipendula (Michx.) Torr.:

E32332; E32374

Dicanthelium oligosanthes (Schult.) Gould:

E32226; E32219 Elmus canadensis L.: E32365 Elymus virginicus L.: E32352 *Poa pratensis L.: E32227; E32220 Schizachyrium scoparium (Michx.) Nash: E32711

*Setariafaberi F.Herrm.: E32377 Sorghastrum nutans (L.) Nash: E32710

Transactions of the Illinois State Academy of Science (2012) Volume 105,#l&2,pp. 19-31

received 10/27/11 accepted 4/22/12

Influence of Culture Conditions on Hydrogen Peroxide Production by Lactobacillus jensenii

Adonica S. Cosgrove Pohren and Scott M. Holt*

Western Illinois University, Department of Biological Sciences 1 University Circle, Macomb, IL 61455, USA Corresponding Author

ABSTRACT

The purpose of this study was to determine how changes in certain culture conditions influence hydrogen peroxide production rate by the vaginal isolate Lactobacillus jensenii. Hydrogen peroxide production was detected for all variables, but the highest rates occurred at 13.8 mM glucose, pH 7, 40°C, and during early logarithmic growth stage when cell density was taken into account. Hydrogen peroxide was produced from L. jen¬ senii regardless of the experimental variable; however, significant differences in produc¬ tion rate were detected within each variable. Since vaginal lactobacilli may play a crucial role in prevention of disease, it is important to know how in vitro environmental factors, which may emulate the vaginal environment or aid in the development of potential vagi¬ nal probiotic strains, influence production of hydrogen peroxide.

INTRODUCTION

Lactobacillus spp., which inhabit mucus membranes of humans and animals, are known to be dominant microorganisms in the reproductive tract of healthy women (Brown, 1978; Hill et al. 1984) and may provide protection from certain vaginal infections (Gupta et al. 1998; Hawes et al. 1996; Martin et al. 1999; Spurbeck and Arvidson, 2008). Disturbance or alteration of the Lactobacillus- dominated vaginal microbiota has been linked to an increased susceptibility to certain infections (Gupta et al. 1998; Hawes et al. 1996; Martin et al. 1999; Wilks et al. 2004). Lactobacillus spp. are thought to maintain or regulate the mucus-membrane microbial ecosystem by production of antimicrobial sub¬ stances such as bacteriocins (Jack et al. 1995), lactic acid, (Boskey et al. 1999; Ogawa et al. 2001) and hydrogen peroxide (Eschenbach et al. 1989, Wilks et al. 2004). Hydrogen peroxide-producing lactobacilli seem to play a crucial role in regulating a healthy vaginal microbiota (Cherpes et al. 2008; Gupta et al. 1998; Hawes et al. 1996; Martin et al. 1999 Wilks et al. 2004). The absence of hydrogen peroxide-producing vaginal Lactobacillus spp. has been associated with an increased risk of bacterial vaginosis (Cherpes et al. 2008; Hawes et al. 1996), certain urinary tract infections (Gupta et al. 1998), and acquisi¬ tion of human immunodeficiency virus (Martin et al. 1999). Wilks et al. (2004) and Cherpes et al. (2008) found that the presence of hydrogen peroxide-producing Lactobacillus spp. in the vagina of pregnant women was linked to a lower risk of bacte-

20

rial vaginosis and uterine infection. Among the numerous Lactobacillus species isolated from the human vagina (L. casei, L. gasseri, L. iners, L. fermentum, L. plantarum, L. brevis , L.delbrueckii , L. vaginalis , L. salivarius), L.jensenii is among the most abundant species (Antonio et al. 1999; Martin et al. 2008; Martin and Suarez, 2010; McGroarty et al. 1992; Vallor et al. 2001; Vasquez et al. 2002). In addition to abundance, nearly all of the vaginal L.jensenii strains reported in the literature make hydrogen peroxide (Antonio et al. 1999; Eschenbach et al. 1989; Martin et al; 2008; Martin and Suarez, 2010; McGroarty et al. 1992; Wilks et al. 2004) and at higher levels than most other vaginal Lactobacillus species (Martin and Suarez, 2010; Wilks et al. 2004). The level of hydro¬ gen peroxide synthesis is an important characteristic of vaginal lactobacilli since there appears to be a relationship between its production and a reduced risk of genitourinary infections (Cherpes et al. 2008; Gupta et al. 1998; Hawes et al. 1996; Martin et al. 1999; Wilks et al. 2004). Therefore, hydrogen peroxide production is an important physiologi¬ cal characteristic for endogenous vaginal lactobacilli and for assessing the potential of a vaginal probiotic strain (Bolton et al. 2008; McLean and Rosenstein, 2000). Since vaginal lactobacilli may play a crucial role in prevention in disease, it is important to know which in vitro environmental factors will influence the rate of hydrogen peroxide production and to what extent. Very little information has been published about the influence of environmental conditions or growth phase on hydrogen peroxide production rate from important vaginal Lactobacillus species such as L.jensenii. A few other studies have also found that certain environmental or culture conditions such as aeration (Martin and Sua¬ rez, 2010; Otero and Nader-Macias, 2006; Tomas et al. 2003), incubation temperature, and culture pH (Tomas et al. 2003) can impact hydrogen peroxide production from vagi¬ nal Lactobacillus isolates. Recently, Martin and Suarez (2010) observed that vaginal L. jensenii cultures grown in media containing Fe3+ did not make hydrogen peroxide. The Fe?+in the growth medium activated a peroxidase made by L.jensenii which degraded the hydrogen peroxide (Martin and Suarez, 2010). It has been previously reported that lactobacilli possess an atypical manganese-containing catalase/peroxidase system (Barynin et al. 2001; Yoder et al. 2000). Considering that lactobacilli possess a cata¬ lase/peroxidase and that complex growth media may contain Fe3+, it is important to remove spent medium components from lactobacilli cells prior to measuring hydrogen peroxide synthesis. Most of the published literature dealing with vaginal lactobacilli, however, measure hydrogen peroxide production directly from spent growth media with¬ out using a wash step. In addition to a wash step, it is also important to measure the rate of hydrogen peroxide synthesis by lactobacilli rather than by end-point determination. Barnard and Stinson (1999) reported that rates of hydrogen peroxide were a more accu¬ rate assessment of the potential in vivo antagonistic ability of lactic acid bacteria rather than a simple end-point determination. As hydrogen peroxide diffuses away from the producing bacterial strain in vivo, the concentration is reduced and loses its killing effectiveness (Barnard and Stinson, 1999). Therefore, a vaginal lactobacillus isolate that can produce hydrogen peroxide at a high rate may be most effective for antagonism. Most of the published literature dealing with vaginal lactobacilli, however, measure hydrogen peroxide by using an end-point determination which is not a true kinetic assess¬ ment. L. jensenii was used for this study because it has been frequently isolated from the vagina of healthy women and most strains are strong producers of hydrogen peroxide (Antonio et al. 1999; Martin et al. 2008; Martin and Suarez, 2010; McGroarty et al. 1991; Wilks et al. 2004; Vallor et al. 2001; Vasquez et al. 2002). L.jensenii is not yet consid¬ ered a probiotic strain and needs to be further evaluated based on the other characteristics

21

mentioned above. The purpose of this study was to determine how changes in glucose concentration, pH, temperature, and stages of growth influence hydrogen peroxide production rates by the important vaginal isolate and potential probiotic species L. jen- senii.

MATERIALS AND METHODS

Hydrogen peroxide assay

The assay for hydrogen peroxide was adapted from Barnard and Stinson (1999). All assay reagents were prepared in sodium phosphate buffer (0.1 M, pH 7.8). The assay con¬ sisted of 20 pi horseradish peroxidase (2 U, HRP, Sigma, St. Louis), 900 pi of 2,2'-azino- bis-(3-ethylbenzthiazoline)-6-sulfonic acid (90 pg, ABTS, Sigma), and 50 pi of cell supernatant. The assay reagents were immediately mixed by inversion in a one ml cuvette and the absorbance (A414nm) was measured using a spectrophotometer (Genesys 20, Thermospectronic, Madison, WI). The spectrophotometer was adjusted to zero absorb¬ ance with a blank which consisted of 920 pi buffer and 50 pi supernatant. Two controls were used for each reading to adjust for background from the enzyme or substrate. The enzyme control consisted of all components of the assay with the exception of ABTS. The substrate control consisted of all components of the assay with the exception of HRP. An additional control (reagent control) was tested with all of the components of the assay except the cell supernatant to insure that the assay was only detecting hydrogen peroxide production, and not background from the buffer. Cells from each trial were frozen and later counted with a Petroff-Hausser Chamber (data not shown). A standard curve of A vs hydrogen peroxide concentration was prepared using Microsoft Excel. The equation of the regression line for the standard curve was y (A414nm) = 0.094x (H202 mM) - 0.0959 (r2 = 0.998) and was used to calculate hydrogen peroxide concentrations from each sample for all experiments. Initial rates of hydrogen peroxide production were used in this study because it is a more accurate assessment of the ability of certain lactic acid bacteria to control competing bacteria (Barnard and Stinson, 1999).

Data analysis

Four main experiments were performed in this study and included determination of the influence of glucose concentration, pH, temperature, and growth stage on hydrogen peroxide formation in L. jensenii. For each experimental variable, a plot of hydrogen peroxide concentration (mM) vs time (min) was prepared and a linear regression line was determined using Microsoft Excel. The slope of this linear regression line was used to determine units of activity for each experimental variable. One unit (U) of activity was defined as one mM of H202 produced per min under the assay conditions. Each variable was performed in triplicate and activity was reported as the mean U ±standard deviation (SD). ANOVA and Tukey multiple comparison tests were used to determine if differ¬ ences existed among activity means within each experiment and were reported as P val¬ ues. All statistical analyses were performed with Systat 8.0 software (SPSS Inc., Chicago, IL).

Bacterial strain, growth conditions, and cell preparation

Lactobacillus jensenii NRRL B-4550 was obtained from the United States Department of Agriculture, Agriculture Research Service Culture Collection in Peoria, IL. The L. jen¬ senii strain used in this study (B-4550 ) was originally isolated from human vaginal dis-

22

charge as reported by Glasser et al. (1970) and is the L.jensenii type strain (Skerman et al. 1980). The strain has been used extensively in research related to the topic of vaginal lactobacilli and hydrogen peroxide (Antonio et al. 1999; Rabe and Hillier, 2003; Song et al. 1999; Spurbeck and Arvison, 2008; Zhong et al. 1998;). For preparation of stock cul¬ tures, isolated colonies were inoculated into Difco™ MRS Broth (10 ml) (Becton, Dickinson, & Co., Sparks, MD) and incubated overnight at 37°C. Cells from the over¬ night cultures were collected by centrifugation, resuspended in 1 ml fresh MRS Broth containing 20% (v/v) glycerol, and were stored frozen at -80°C until needed. For each experiment, a single vial of frozen stock cells was thawed, gently mixed, and 50-ul was inoculated into the 10 ml MRS broth. Each spent frozen stock culture was not re-frozen but instead destroyed. A new vial of frozen stock culture was used for each experiment. This approach provided for very consistent inocula, much more consistent than perform¬ ing multiple cultivations (two or three cultivations or transfers) prior to the assay. The inoculum provided consistent results for lag, log, and stationary phases and for enumerat¬ ing cell numbers by microscopic cell count using Petroff-Hausser Chamber before each hydrogen peroxide assay was performed. The growth curve showed the expected lag, log, and stationary phases (Fig 1). The inoculated test tube culture (10 ml MRS broth) was subsequently incubated overnight at 37°C. Growth of the culture was measured by A600nm using a spectrophotometer (Spectronic 21, Bausch and Lomb). Cells were collected by centrifugation to a pellet (Sorvall angle centrifuge, Sorvall L.L.C., New Castle, DE) and washed three times with sodium phosphate buffer (0.1 M, pH 7.8, Breznak and Costilow, 1994) to remove medium components and glucose. The cells were resuspended in fresh phosphate buffer and adjusted to a A600 value of 1.0. This prepared culture was then used to determine the influence of environmental factors (glucose, pH, and temperature) on hydrogen peroxide production.

Influence of glucose on hydrogen peroxide production by L.jensenii

Samples (200 pi) from a prepared culture were distributed into each of five pre-warmed (37°C water bath) microcentrifuge tubes containing 200 pi of 0.1 M sodium phosphate buffer (pH 7.8) and varying concentrations of glucose (mM). The lids were closed and the tubes were incubated for up to 40 min at 37°C. One tube was removed from the water bath at each time interval (0, 10, 20, 30, 40 min). At each time interval, the tube was centrifuged for three min at 14,000x g (Eppendorf 5415C, Westbury, NY) to prepare a clarified supernatant, which was assayed for hydrogen peroxide.

Influence of pH on hydrogen peroxide production by L.jensenii

Cell preparation and the assay conditions were the same as previously described except that glucose concentration was maintained at 13.8 mM and the pH of the test buffers var¬ ied. The buffers used to test the effects of pH included citrate-phosphate buffer (pH 7 and pH 6) and acetate buffer (pH 5 and pH 4).

Influence of temperature on hydrogen peroxide production by L.jensenii

Cell preparation and assay conditions were the same as previously described except that glucose concentration was maintained at 13.8 mM, pH was maintained at 7.0, and the incubation temperatures were 30°C, 35°C, 40°C, and 45°C.

23

Influence of L.jensenii growth stage on hydrogen peroxide production.

A growth curve was prepared (data not shown) by inoculation of a L.jensenii thawed stock culture (65 pi) into a 16 -mm screw-cap test tube containing 13 ml of MRS broth which was incubated at 37°C in a water bath. Growth (A600nm) was measured using a spectrophotometer (Spectronic 21) at three-h intervals for 15 h, and again at 20 h. Hydro¬ gen peroxide formation was immediately detected from each sample collected during the growth assessment as described previously. The cells were washed and resuspended in fresh phosphate buffer using optimum conditions (13.8 mM, glucose, pH 7). Samples were assayed during early logarithmic (3 h), late logarithmic (6 h), early stationary (9 h), and late stationary phase (15 h) of growth.

RESULTS

Hydrogen peroxide was produced regardless of the glucose concentrations tested (Figure 2). The highest rate of hydrogen peroxide production occurred at 13.8 mM glucose (0.27 U ± 0.08) and the lowest rate was at 55.5 mM glucose with an activity of 0.06 U ±0.01 (Figure 2). Hydrogen peroxide production at 55.5 mM glucose and when no glucose was added to the assay was significantly different than at 13.8 mM glucose (P = 0.031, P = 0.003, respectively) (Figure 2). In addition, hydrogen peroxide production at 55.5 mM glucose was significantly different from 27.7 mM glucose and 41.6 mM glucose (P = 0.028, P = 0.025) (Figure 2).

Hydrogen peroxide was produced regardless of pH conditions tested (Figure 3). The highest rate of hydrogen peroxide production was detected at pH 7, with an activity of 0.3 1U ± 0.09 and the lowest rate of production was at pH 5 (0.07 U ± 0.00) (Figure 3). There was a significant difference in hydrogen peroxide production between pH 7 and pH 5 (P = 0.004), pH 7 and pH 4 (P = 0.021), and pH 5 and pH 6 (P = 0.020) (Figure 3).

Hydrogen peroxide was produced regardless of the assay temperatures tested (Figure 4). The highest rate of hydrogen peroxide production occurred at 40°C (0.19 U ±0.01) and the lowest rate of production occurred at 30°C (0.05 U ±0.01) (Figure 4). A significantly lower rate of hydrogen peroxide production occurred at 30°C when compared to the other temperatures tested (35°C, P = 0.003; 40°C, P = 0.000, and 45°C, P = 0.000) (Figure 4). A significant difference in hydrogen peroxide production also occurred between 40°C and 35°C (P = 0.003) (Figure 4).

The relationship between growth stage and hydrogen peroxide production by L.jensenii is shown in figures 5 and 6. Hydrogen peroxide production was calculated as mM L 1 min'1 (U) in figure 5 and as mM L 1 min 1 / A in figure 6. Hydrogen peroxide assays calculated as mM L'1 min'1 / A value were determined to account for differences in hydrogen peroxide production due to cell density (Figure 6). Hydrogen peroxide was produced regardless of the growth stages tested (three h, mid log; six h, late log; nine h, early stationary; and 15 h, late stationary, Figures 5 and 6). When calculated as mM L 1 min'1, the highest rate of hydrogen peroxide production occurred after six h growth (late log phase, 0.24 U ± 0.05) and the lowest rate occurred after three h growth (mid log phase, 0.07 U ± 0.02) (Figure 5). A significant difference in hydrogen peroxide produc¬ tion occurred between three h and six h growth (P = 0.0002), six h and nine h growth (P = 0.024), and three h and 15 h growth (P = 0.007) (Figure 5). When cell density was

24

taken into account as mM L 1 min'1 / A, the highest hydrogen peroxide production occurred at three h growth (mid log phase, 1.11 ± 0.35) and the lowest activity occurred at nine h growth (early stationary phase, 0.10 ±0.01) (Figure 6). A significant difference in hydrogen peroxide production occurred between three h and six h growth (P = 0.006), three h and nine h growth (P = 0.002), and three h and 15 h growth (P = 0.002) (Figure 6).

DISCUSSION

The purpose of this study was to determine how certain in vitro culture conditions influ¬ ence the rate of hydrogen peroxide production by L.jensenii. It was apparent from this study that many factors can influence in vitro hydrogen peroxide production by L. jen- senii. For example, hydrogen peroxide production by L.jensenii was lowest at the highest glucose concentration (55.5 mM) and at the lowest glucose concentration (0 mM) tested. These data were similar to findings by Barnard and Stinson (1999) who studied hydrogen peroxide synthesis in Streptococcus gordonii. S. gordonii is a lactic acid bacterial species associated with oral health. Barnard and Stinson (1999) showed that there was essentially a bell shaped curve associated with hydrogen peroxide production as it related to glucose concentration. In their study, hydrogen peroxide synthesis was lowest at the lowest (0.01 mM) and highest (1000 mM) glucose concentrations tested and was higher at the intermediate glucose concentrations (O.lmM, 1.0 mM, and 10 mM). The fact that both L. jensenii (in this study) and S. gordonii (Barnard and Stinson (1999) displayed lower hydrogen peroxide synthesis at the highest glucose concentrations tested in each study may reflect a common mechanism used by lactic acid bacteria to efficiently regulate bacterial competition in a vaginal or oral environment. Competition for nutrients among various microbial groups in the oral or vaginal environment would be less important when carbohydrate concentration is abundant. Consequently, a high rate of hydrogen peroxide synthesis by lactic acid bacteria is not needed to suppress competing bacteria and the cells can devote resources to more important metabolic processes. L.jensenii and S. gordonii (Barnard and Stinson 1999) both produced hydrogen peroxide when glucose was not added to the assay conditions. The energy needed for hydrogen peroxide synthe¬ sis by lactic acid bacteria in the absence of extraneous glucose may have come from metabolism of intracellular polysaccharide stores (Barnard and Stinson, 1999; Minah and Loesche, 1977). Minah and Loesche (1977) showed that up to 14% of the carbohydrate consumed by hydrogen peroxide-producing oral streptococci is converted into intracellu¬ lar carbohydrate. The ability of L. jensenii to produce an intracellular polysaccharide reserve, however, was not determined in this study. Although not statistically significant, L. jensenii made twice as much hydrogen peroxide with no exogenous glucose (0 mM) than with the highest glucose concentration tested (55.5 mM). The ability to make hydro¬ gen peroxide without exogenous carbohydrate present may reflect the need for certain lactic acid bacteria (L. jensenii or S. gordonii , Barnard and Stinson, 1999) to inhibit competing bacteria in their particular environment (vaginal or oral) under low carbohy¬ drate conditions when competition for nutrients would be more intense (Barnard and Stinson, 1999). Vaginal glucose in the form of glycogen (Gregoire et al. 1971; Paavonen, 1983) is used by lactic acid bacteria as an energy source for hydrogen peroxide synthesis (Barnard and Stinson, 1999; Minah and Loesche, 1977) and is also metabolized to form lactic acid (Boskey et al. 2001). Glycogen content will vary based on the cycle stage and

25

vaginal location but has been determined to be 1122 jiig to 1667 jug of glycogen per 100 mg of tissue wet weight epithelial tissue (Gregoire et al. 1971).

Vaginal glycogen levels are regulated by estrogen (Gregoire et al. 1971; Paavonen, 1983). Consequently, fluctuations in estrogen such as menarche, menopause, oral contraception, and hormone replacement therapy could conceivably alter the vaginal biota including hydrogen peroxide-producing lactobacillus species (Cauci et al. 2002). Increased estrogen production can cause higher amounts of glycogen to be deposited into the vagina (Boskey et al. 2001; Gregoire et al. 1971; Paavonen, 1983). It is hypothesized that decreased vaginal glycogen levels may increase the antimicrobial competitive activi¬ ties (lactic acid and hydrogen peroxide production) of vaginal lactobacilli.

Production of hydrogen peroxide by L.jensenii in this study was also influenced by pH. The higher production rates of hydrogen peroxide detected in this study at pH 6 and pH 7 than at pH 4 and 5 may be a response by L. jensenii to inhibit microbial competition in the vaginal environment as the pH increases. The healthy vagina should have a pH range of 4. 0-4 .7 (Gardner and Dukes 1955) and an abundance of the lactobacilli morphotype (large gram-positive rod) (Nugent et al. 1991; Spiegel, 1991). An increase in vaginal pH above 4.5, a decrease in lactobacilli, and the appearance of non lactobacilli bacterial mor- photypes (gram-negative rods, gram- variable rods and curved rods, gram positive cocci) are among the clinical signs associated with bacterial vaginosis (Amsel et al. 1983; Nugent et al. 1991; Spiegel, 1991). The lowest rates of hydrogen peroxide synthesis obtained in this study were detected at pH 4 and pH 5. This response to pH may be a mechanism for L.jensenii to conserve energy during times of lower pH because the nor¬ mal acidic vaginal environment (pH 4. 0-4 .7) itself would help inhibit the growth of pathogenic organisms (Ogawa et al. 2001; Stamey and Timothy, 1975).

Temperature was also an important factor in the production of hydrogen peroxide by L. jensenii. The peak rates of production (0.19 mM L"1 min"1 and 0.16 mM L"1 min-1) occurred at the two highest temperatures tested (40°C and 45°C), indicating that increased temperature enhances the production of hydrogen peroxide in L. jensenii in vitro. Although the difference in rates between the two highest temperatures was not significant, cells assayed for hydrogen peroxide production at 45°C may have been slightly temperature stressed resulting in the lower rate. In general, higher hydrogen peroxide synthesis rate coordinated with optimum growth temperatures (37°C-44°C) associated with Lactobacillus species isolated from the vagina (McLean and Rosenstein, 2000; Tomas et al. 2003). The coordination of high hydrogen peroxide synthesis with optimum growth temperatures that mimic the human vagina (37°C) is an important physiological characteristic for endogenous vaginal lactobacilli and for assessing the potential of a vaginal probiotic strain (McLean and Rosenstein, 2000).

In addition to the other environmental factors mentioned in this study, hydrogen peroxide production by L.jensenii coordinated with growth stage (Fig 6) rather than by cell num¬ bers alone (Fig 5). That is, actively growing cells of L.jensenii and not necessarily high cell numbers produced the highest rates of hydrogen peroxide. For example, when meas¬ ured as mM L 1 min'1, hydrogen peroxide synthesis was assessed by cell numbers during the growth cycle (Fig 5). For this cell number assessment, hydrogen peroxide synthesis was lowest during mid-log (0.07 mM L 1 min'1) but peaked during late-log growth phase

26

(0.24 mM L"1 min'1). When calculated as mM L 1 min'1 / A value, hydrogen peroxide synthesis peaked during mid-log growth (1.11 mM L1 min"1 / A) followed by a sharp reduction in activity as the growth cycle proceeded into late log (0.29 mM L 1 min'1 / A) and stationary phase (0.10 mM L_1min 1 / A; 0.13 mM L 1 min"1 / A) (Fig 6). This activity assessment (mM L'1 min 1 / A value) is an indication of hydrogen peroxide synthesis effi¬ ciency by L. jensenii. This value may be a more accurate assessment of the hydrogen peroxide capability of a probiotic culture since lactobacilli cell numbers in the healthy vagina (Boskey et al. 1999) would not reach the concentration or density as is typically obtained in vitro (Tomas et al. 2003). In addition, these data may also indicate that hydro¬ gen peroxide production in L. jensenii coordinates with the synthesis of cell enzymes needed for primary metabolism such as NADH oxidases (Marty-Tysset et al. 2000; Talwalkar and Kailasapathy, 2003).

This in vitro study provides the first evidence that hydrogen peroxide synthesis rate by the vaginal isolate L. jensenii is not static but is highly influenced by environmental fac¬ tors. L. jensenii can adjust hydrogen peroxide synthesis rate in response to changes in glucose concentration, pH, temperature, and growth phase. Parameters such as glucose concentration and pH are important environmental conditions that can influence composi¬ tion of the vaginal microbiota. It is not ideal to correlate results of an in vitro study to an in vivo environment such as the vagina. The response in hydrogen peroxide synthesis by L. jensenii to glucose and pH fluctuations, however, may reflect a common mechanism used by lactic acid bacteria to efficiently regulate bacterial competition in the vaginal environment. This study may help to establish a scientific basis for the selection of vagi¬ nal probiotic bacterial strains and also may be used as a guideline for studies focused on differential gene expression in response to changes in environmental conditions.

LITERATURE CITED

Amsel, R. Totton, P.A., Spiegel, C.A., Chen, K.C.S., Eschenbach, D.A., and Holmes, K.K. (1983) Nonspecific vaginitis: diagnostic criteria and microbial and epidemiological associations. Am J Med 74:14-22.

Antonio, M.A.D., Hawes, S.E., and Hillier, S.L. (1999) The identification of vaginal Lactobacillus species and the demographic and microbiologic characteristics of women colonized by the spe¬ cies. J Infect Dis 180:1950-1956.

Barnard, J.P. and Stinson, M.W. (1999) Influence of environmental conditions on hydrogen perox¬ ide formation by Streptococcus gordonii. Infect Immun 67:6558-6564.

Barynin, V.V., Whittaker, M.M., Antonyuk, S.V., Lamzin, V.S., Harrison, P.M., Artymiuk, P.J. and Whittaker, J.W. (2001) Crystal structure of manganese catalase from Lactobacillus planta- rum. Structure 9:725-738.

Bolton M., Van Der Straten, A. and Cohen, C.R. (2008) Probiotics: potential to prevent HIV and sexually transmitted infections in women. Sex Trans Dis 35:214-225.

Boskey, E.R., Telsch, K.M., Whaley, K.J., Moench, T.R. and Cone, R.A. (1999) Acid production by vaginal flora in vitro is consistent with the rate of extent of vaginal acidification. Infect Immunology, 67:5170-5175

Boskey, E.R., Cone, R. A., Whaley, KJ. and Moench, T.R. (2001) Origins of vaginal acidity: high D/L lactate ratio is consistent with bacteria being the primary source. Hum Reprod 16:1809- 1813.

Breznak, J.A. and Cosilow, R.N. (1994) Physicochemical factors in growth. In: Gerhardt, P., Mur¬ ray, R.G.E., Wood, W.A. and Krieg N.R. (ed), Methods for General and Molecular Bacteriology, Washington, D.C.: American Society for Microbiology, pp. 137-154.

Brown, W.J. (1978) Microbial ecology of the normal vagina. In Hafez, E.S.E. and Evans, T.N. (ed), The Human Vagina, North-Holland Publishing Co., Amsterdam, pp. 407-422.

27

Cauci, S., Driussi, S., De Santo, D., Pencchioni, P., Iannicelli, T., Lanzafame, P., De Seta, F., Quadrifoglio, F., deAloysio, D and Guaschino, S. (2002) Prevalence of bacterial vaginosis and vaginal flora changes in peri- and postmenopausal women. J Clin Microbiol 40:2147-2152.

Cherpes, T.L., Hillier, S.L., Meyn, L.A., Busch, J.L. and Krohn, M.A. (2008) A delicate balance: risk factors for acquisition of bacterial vaginosis include sexual activity, absence of hydrogen peroxide-producing lactobacilli, black race, and positive herpes simplex virus type 2 serology. Sex Transm Dis 35:78-83.

Eschenbach, D.A., Davick, P.R., Williams, B.L., Klebanoff, S.J., Young-Smith, K., Critchlow, C.M., and Holmes, K.K. (1989) Prevalence of hydrogen peroxide-producing Lactobacillus spe¬ cies in normal women and women with bacterial vaginosis. J Clin Microbiol 27:251-256.

Gardner, H.L and Dukes, C.D. (1955) Heamophilus vaginalis vaginitis. A newly defined specific infection previously classified “nonspecific” vaginitis. Am J Obstet Gynecol 69:962-976.

Gasser F, Mandel, M, and Rogosa, M. (1970) Lactobacillus jensenii sp. nov., a new representative of the subgenus Thermobacterium. J Gen Microbiol 62:219-222.

Gregoire, A.T., Kandil, O. and Ledger, W.J. (1971) The glycogen content of human vaginal epithe¬ lial tissue. Fertil Steril 22:64-68.

Gupta, K., Stapleton, A.E., Hooton, T.M., Roberts, P.L., Fennell, C.L. and Stamm, W.E. (1998) Inverse association of H202-producing lactobacilli and vaginal Escherichia coli colonization in women with recurrent urinary tract infections. J Infect Dis 178:446-450.

Hawes, S.E., Hillier, S.L., Benedetti, J., Stevens, C.E., Koutsky, L.A., Wolner-Hanssen, P. and Holmes, K.K. (1996) Hydrogen peroxide-producing lactobacilli and acquisition of vaginal infec¬ tions. J Infect Dis 174:1058-1063.

Hill, G.B., Eschenbach, D.R. and Holmes, K.K. (1984) Bacteriology of the vagina. Scand J Urol Nephrol Suppl 86:23-39.

Jack, R.W., Tagg, J.R. and Ray, B. (1995) Bacteriocins of gram-positive bacteria. Microbiol Rev 59:171-200.

Martin, H.L., Richardson, B.A., Nyange, P.M., Lavreys, L., Hillier, S.L., Chohan, B., Mandaliya, K., Ndinya-Achola, J.O., Bwayo, J. and Kreiss, J. (1999) Vaginal lactobacilli, microbial flora, and risk of human immunodeficiency virus type 1 and sexually transmitted disease acquisition. J Infec Dis 180:1863-1868.

Martin, R., Soberon, N., Vaneechoutte, M., Florez, A.B., Vazquez, F. and Suarez, J.E. (2008) Characterization of indigenous vaginal lactobacilli from healthy women as probiotic candidates. Int Microbiol 1 1:261- 266.

Martin, R., and Suarez, J.E. (2010) Biosynthesis and degradation of H202 by vaginal lactobacilli. Appl Environ Microbiol 76:400-405.

Marty-Teysset, C., de la Torre, F. and Garel, J.R. (2000) Increased production of hydrogen perox¬ ide by Lactobacillus delbrueckii subsp. bulgaricus upon aeration: involvement of an NADH oxi¬ dase in oxidative stress. Appl Environ Microbiol 66:262-267 .

McGroarty, J.A., Tomeczek, L., Pond, D.G., Reid, G. and Bruce, A.W. (1992) Hydrogen peroxide production by Lactobacillus species: correlation with susceptibility to the spermicidal compound nonoxynol-9. J Infect Dis 165:1 142-1 144.

McLean, N.W. and Rosenstein, I.J. (2000) Characterization and selection of a Lactobacillus species to re-colonize the vagina of women with recurrent bacterial vaginosis. J Med Microbiol 49:543- 552.

Minah, G.E. and Loesche, W.J. (1977) Sucrose metabolism by prominent members of the flora isolated from cariogenic and non-cariogenic dental plaques. Infect Immun 17:55-61 .

Nugent, R.P., Krohn, M.A. and Hillier, S.L. (1991). Reliability of diagnosing bacterial vaginosis is improved by a standardized method of gram stain interpretation. J Clin Microbiol 29:297-301.

Ogawa, M., Shimizu, K., Nomoto, K., Tanaka, R., Hamabata, T., Yamasaki, S., Takeda, T. and Takedam, Y. (2001) Inhibition of in vitro growth of Shiga toxin-producing Escherichia coli 0157:H7 by probiotic Lactobacillus strains due to production of lactic acid. Int J Food Microbiol 68:135-140.

Otero, M. C. and Nader-Macias, M.E. (2006) Inhibition of Staphylococcus aureus by H202-produc- ing Lactobacillus gasseri isolated from the vaginal tract of cattle. Anim Reprod Sci 96:35-46.

Paavonen, J. (1983) Physiology and ecology of the vagina. Scand J Infect Dis Suppl 40:31-35.

Rabe, L.K. and Hillier, S.L. (2003) Optimization of media for detection of hydrogen peroxide production by Lactobacillus species. J Clin Microbiol 41:3260-3264.

28

Skerman, V.B.D., McGowan, V., and Sneath, P.H.A. (1980) Approved lists of bacterial names. Int. J. Syst. Bacteriol. 30:225-420.

Song, Y.L, Kato, N., Matsumiya, Y., Liu, C.X., Kato, H. and Watanabe, K. (1999) Identification of and hydrogen peroxide production by fecal and vaginal lactobacilli isolated from Japanese women and newborn infants. (1999) J Clin Microbiol 37:3062-3064.

Spiegel, C.A. (1991) Bacterial vaginosis. Clin Microbiol Rev 4:485-502.

Spurbeck, R.R. and Arvidson, C.G. (2008) Inhibition of Neisseria gonorrhoeae epithelial cell interactions by vaginal Lactobacillus species. Infect Immun 76:3124-3130.

Stamey, T.A. and Timothy, M.M. (1975) Studies of introital colonization in women with recurrent urinary infections. I. The role of vaginal pH. J Urol 1 14:262-263.

Talwalkar, A. and Kailasapathy, K. (2003) Metabolic and biochemical responses of probiotic bacte¬ ria to oxygen. J Dairy Sci 86:2537-2546.

Tomas, M.S., Bru, E. and Nader-Macias, M.E. (2003) Comparison of the growth and hydrogen peroxide production by vaginal probiotic lactobacilli under different culture conditions. Am J Obstet Gynecol 188:35-44.

Vallor, A.C., Antonio, M.A.D., Hawes, S.E. and Hillier, S.L. (2001) Factors associated with acquisition of, or persistent colonization by vaginal Lactobacilli: role of hydrogen peroxide production. J Infect Dis 184:1431-1436.

Vasquez A., Jakobsson, T., Ahme, S., Forsum, U. and Molin, G. (2002) Vaginal Lactobacillus flora of healthy Swedish women. J Clin Microbiol 40:2746-2749.

Wilks, M., Wiggins, R., Whiley, A., Hennessy, E., Warwick, S., Porter, H., Corfield, A. and Millar, M. (2004) Identification and H202 production of vaginal Lactobacilli from pregnant women at high risk of preterm birth and relation with outcome. J Clin Microbiol 42:713-717.

Yoder, D.W., Hwang, J. and Penner-Hahn, J.E. (2000) Manganese catalases. Met Ions Biol Syst 37:527-557.

Zhong, W., Millsap, K., Bialkowska-Hobrzanska, H., and Reid, G. (1998) Differentiation of Lactobacillus species by molecular typing. Appl Environ Microbiol 64:2418-2423.

29

Figure 1. Growth curve of L. jensenii after inoculation into a 10 ml MRS broth culture.

The 10-ml MRS culture was started from a frozen stock culture (50 ul) of L. jensenii. Each time point represents data from three growth trials. Absorbance values were exactly the same for each time point.

Growth of Lact. jensenii in MRS Using Frozen

Stock as Inoculum

Time of Growth (hours)

Figure 2. Influence of glucose concentration on hydrogen peroxide production by L. jen¬ senii. Hydrogen peroxide production rate (mM L'1 min1) was determined from three trials for each glucose concentration and reported as mean ± SD.

0.35

0.30

0.25

0.20

0.15

0.10

0.05

0.00

0 13.8 27.7 41.6 55.0

Glucose Concentration (mM)

Hydrogen Peroxide (mM L1 min*1) w Hydrogen Peroxide (mM L 1 min1)

30

Figure 3. Influence of pH on hydrogen peroxide production by L. jensenii. Hydrogen peroxide production rate (mM L 1 min'1) was determined from three trials for each pH and reported as mean ± SD.

4 5 6 7

pH

ire 4. Influence of temperature on hydrogen peroxide production by L. jensenii. Hydrogen peroxide production rate (mM L'1 min _1) was determined from three trials for each temperature and reported as mean ± SD.

30°C

35°C 40°C

T emperature

-

*

45 °C

31

Figure 5. Influence of L. jensenii growth stage on hydrogen peroxide production meas¬ ured as mM L 1 min"1. Hydrogen peroxide production rate (mM L 1 min"1) was determined in triplicate for each growth stage and reported as mean ± SD.

c 0.3

I

I I 0.25 ■

mid-log late log early station late station

3 h 6 h 9 h 15 h

Stages and Hours of Growth

Figure 6. Influence of L. jensenii growth stage on hydrogen peroxide production meas¬ ured as (mM L 1 min"1 / A). Hydrogen peroxide production rate (mM L"1 min'1 / A) was determined from three trials for each growth stage and reported as mean ± SD.

mid-log late log early station late station 3 h 6 h 9 h 15 h

Stages and Hours of Growth

Transactions of the Illinois State Academy of Science (2012) Volume 105, #1&2, pp. 33-39

received 6/24/1 1 accepted 2/5/12

Prevalence of Baylisascaris procyonis and Implications for Reintroduced Woodrat Populations in Southern Illinois

Simon R. Bade, F. Agustfn Jimenez, Aaron K. Poole, and George A. Feldhamer Department of Zoology, Southern Illinois University, Carbondale, IL 62901-6501

ABSTRACT

The roundworm Baylisascaris procyonis (Nematoda: Ascaridae) is carried and spread by raccoons ( Procyon lotor ). In intermediate hosts the parasite can cause brain damage and death. The prevalence of B . procyonis in raccoon populations is a concern in areas where the state-endangered eastern woodrat ( Neotoma floridana ) occurs or has been reintro¬ duced because the foraging and hoarding strategies of woodrats (“packrats”) make them particularly susceptible to infection. To determine the prevalence of this parasite and the possible implications for reintroduced eastern woodrats, we established 10 transects from Garden of the Gods (Saline County) east to Rim Rock/Pounds Hollow (Gallatin County) where woodrats have been reintroduced in Illinois. Raccoon latrine sites were visually located along these transects and scat piles sampled and analyzed using Fecalyzer flota¬ tion kits. Of 79 total fecal samples, only 1 tested positive for B. procyonis. This low prevalence (1.3%) for B. procyonis suggests that it likely poses little risk to the reintro¬ duced woodrat populations within the study area.

Key words: Baylisascaris procyonis , eastern woodrats, raccoons, reintroductions

INTRODUCTION

Raccoons ( Procyon lotor ) have a wide geographic distribution in North America and population densities are often quite high (Gehrt 2003). They thrive in proximity to humans and low pelt prices and reduced trapping effort have increased their abundance. Their large population sizes and ubiquitous distribution make raccoons an important vec¬ tor for parasites and diseases including distemper, rabies, and the roundworm Baylisasca¬ ris procyonis (Mitchell et al. 1999).

Raccoons develop communal latrine sites used by various individuals (Gehrt 2003, Page et al. 2009a, b). Rodents often forage on scats at latrine sites for undigested seeds (LoGiudice 2001). This behavior presents a strong potential method of transmission of B. procyonis (Page et al. 1998). The parasite has little to no effect on the raccoon host and is rarely fatal except in cases of severe infection (Kazacos 2001, Gehrt 2003). In an acci¬ dental host infected with the parasite, the larvae migrate to ocular and brain tissue, encyst there and can cause listlessness, emaciation, loss of motor skills, coma, and ultimately death (Kazacos et al. 1984).

34

The prevalence of raccoons and their latrine sites may have a direct impact on other wild¬ life species, including woodrat populations. Whereas eastern woodrats ( Neotoma flori- dana) once ranged throughout much of southern Illinois, today only four remnant popula¬ tions remain. These show evidence of inbreeding (Monty et al. 2003) and the species is listed as endangered in the state. Reintroduction efforts in the Shawnee Hills region began in 2002 (Feldhamer and Poole 2008), with woodrats from Arkansas and Missouri released in formerly occupied sites at Garden of the Gods, Buzzard’s Point, High Knob, and Pounds Hollow/Rim Rock. Reasons for the decline of woodrats in Illinois are unknown, but may have included habitat loss, degradation, and fragmentation; predation; competition; and mortality due to parasitism from B . procyonis (Birch et al. 1994; Feld¬ hamer and Poole 2008). A high incidence of B. procyonis could have a negative impact on population viability of reintroduced woodrats in Illinois because the foraging strate¬ gies of eastern woodrats make them particularly susceptible to infection. Woodrats rou¬ tinely collect and store items from their habitat, including raccoon scat. Consumption of fresh B . procyonis eggs is not harmful because they are not infective until they embryo- nate in 2 to 4 weeks. Woodrats, however, store scats and may not eat the seeds, giving eggs time to become infective (Page et al. 1999). Furthermore, the eggs remain viable for several years, creating the possibility for increased risk of infection as the bank of eggs increases.

The primary objective of this study was to determine the prevalence of B. procyonis in the local raccoon population inhabiting areas of woodrat reintroductions. The secondary objective was to determine significant microhabitat variables associated with location of raccoon latrines.

MATERIALS AND METHODS

Study Area

Between 2002 and 2009, 422 woodrats were reintroduced into 4 sites within the Shawnee National Forest, from Garden of the Gods (Saline County) east to Buzzard’s Point, High Knob, and Rim Rock (Gallatin County) (Figure 1). These sites were selected for reintroduction based on documented past presence of the species and their bluff escarp¬ ments that provide rocky habitat (see Novosak 2004). Dominant overstory is oak ( Quer - cus spp.)-hickory ( Carya spp.) forest, although Rim Rock is primarily beech ( Fagus spp.)-maple ( Acer spp.) forest. The forest above the bluffs is xeric, while below the bluffs the habitat is more mesic. Self-sustaining populations of woodrats have been established at these sites (Feldhamer and Poole 2008, Ing 2008).

Latrine/Scat Analysis

We established 10 1-km transects between Garden of the Gods and Pounds Hollow to sample raccoon latrine sites. Five transects were on the upland slopes of the bluff line, where reintroduced woodrats may have dispersed, and 5 were below the bluff line in areas known to harbor reintroduced woodrats. Transects were unbounded and all latrines that could be located visually were sampled. Likely sites for raccoon latrines (downed logs and prominent rocks) were examined along the route, flagged, and a GPS location determined. Each identified site constituted an individual sampling unit. Size of latrines was classified as small (1 scat), medium (2 scats), or large (>3 scats) based on amount of

35

scat. Number of samples taken was dependent upon the size of each scat pile — small piles were sampled once, medium piles twice, and large piles three times. Sampling began March 2008, and a circuit of all 10 transects was made monthly for 20 months except August 2008. Different transects were visited weekly, with even effort among all transects. During June and July 2008, transects were walked twice each month. Checking continued through October 2009 for a total of 210 km monitored.

A sample of about 3 g was collected from individual scats in each latrine. Samples were stored at -70° C. Presence/absence of B . procyonis ova was determined by flotation using sodium nitrate and the Fecalyzer system (Evsco Pharmaceuticals, Bueno, NJ) following manufacturers recommendations. Eggs were identified using guidelines for morphology and size from Page et al. (2005). Eggs of B . procyonis are oval-shaped, generally between 65 and 80 //m in length, and have thick, granulated shells. They differ from eggs of Toxo- cara spp. in shape, membrane thickness and appearance (Kazacos and Boyce 1989).

Habitat Measurements

Microhabitat variables were measured at each latrine site and included: number of downed trees (coarse woody debris) in a 5-m radius surrounding the latrine, substrate (bare stone, log, or soil), and distance to nearest tree > 30 cm diameter at breast height (DBH). These habitat variables were also collected at random sites by choosing a cardinal direction at each latrine site and pacing 50 steps in that direction.

Statistical Analyses

We used a 1 -tailed, paired t-test to determine differences in the amount of coarse woody debris and distance to the nearest tree at latrine sites vs. random sites. Chi-square anal¬ yses were used to test for differences in location of latrine sites above or below bluff line, and on substrate types. Level of significance was a = 0.05.

RESULTS

We collected 79 fecal samples from 54 latrine sites. Most of the scat found was in single piles. However, raccoons used several latrine sites repeatedly during the course of the sampling period. Thus, there were only 40 unique latrine sites, or 1 for every 5.25 km monitored. Of the 79 samples, only 1 (1.3%) — from Garden of the Gods — contained eggs of B . procyonis .

Four substrate types contained latrines: rock (n = 18), downed log (n = 13), leaf litter (n = 8), and live tree (n = 1). There was a significant difference among substrate types for occurrence of latrines (x2 = 15.8, df = 3, P < 0.05). Essentially the same substrate types occurred on the 40 randomly selected sites as the identified latrine sites, with the excep¬ tion of one random site with grass substrate. The mean amount of coarse woody debris at latrine sites (28.12%) was greater (t39 = 2.46, P < 0.009) than at random sites (16.62%). The mean distance to a tree > 30 cm DBH was 5.08 m at the latrine sites and 5.05 m at the random sites; these were not significantly different (t39 = 0.72, P = 0.473).

Considering location of the 40 latrines with respect to the bluff line, there were 20 above and 20 below. However, when each woodrat release site was considered independently, there were significant differences. At Garden of the Gods, there were 13 latrine sites

36

above the bluff line and only 3 below (%2 = 6.5, df = 1, P < 0.05). At Rim Rock, there were 3 latrine sites above the bluff line and 10 below (%2 = 3.76, df = 1, P < 0.05). Sam¬ ple sizes at High Knob (3 above and 4 below) and Buzzard’s Point (2 above and 2 below) were too small to test statistically.

Seasonal variation in scat presence was apparent. Scat was most frequently detected in autumn [Sep.-Nov.] ( X= 4.2 scats/month), winter [Dec.-Feb.] ( X - 6.3 scats/month), and spring [Mar.-May] (X= 5.3 scats/month), while an average of only 0.6 scats/month was found during June, July, and August.

DISCUSSION

Occurrence of Baylisascaris procyonis

Previous studies have determined the prevalence of infection of B . procyonis among rac¬ coon populations in southern Illinois. Birch et al. (1994) found only 5.0% prevalence of B. procyonis among a sample of 60 raccoons in southern Illinois. This was much lower than the 64% prevalence found by Barnstable and Dyer (1974) in a sample of 36 raccoons. A more recent study by Nielsen et al. (2007) at Union County Conservation Area (UCCA) — with a very high density of raccoons — found B. procyonis at 16% of latrine sites. Prevalence of B. procyonis is positively correlated with the density of raccoons (Gompper and Wright 2005) and increased raccoon contact rates lead to increased rates of internal parasitism (Wright and Gompper 2005). Anthropogenic impacts that increase resources also can increase parasite prevalence, as can age structures of raccoon popula¬ tions (Prange et al. 2003, Page et al. 2009b).

Given the well-documented ubiquitous nature of raccoons (Gerht 2003, Nielsen et al. 2007), we expected that scat would be abundant and easy to find on our study area. The number of scats we found was much lower than expected, however, and may indicate a low density of raccoons. The study area is removed from most development and the sites generally have little human use. Garden of the Gods is the only site that permits camping; Rim Rock has extensive hiking and picnic use, whereas High Knob and Buzzard’s Point are more remote and less accessible to people with only hiking and equestrian use permit¬ ted. Locations of scats relative to the bluff line paralleled human activity. At Garden of the Gods most use is above the bluff line, whereas at Rim Rock picnic tables are only below the bluff line. High Knob has mostly equestrian use below the bluff line, whereas few people visit Buzzard’s Point.

Raccoons are well known for their tendency to gravitate toward urban and suburban areas for easier access to food, with densities often higher than in rural areas (Prange et al. 2003). The study site at Garden of the Gods had the highest number of latrines and also the most human use. Likewise, UCCA experiences higher human use for camping, boat¬ ing, fishing, and hiking, and high densities of raccoons; Nielsen et al. (2007) reported 1.67 raccoons/ha there. The apparent seasonal variation in scat prevalence we found, with fewer scats during summer, also was unexpected. Whether this was because of seasonal variation in raccoon habitat, increased decomposition rate, increased scat foraging by rodents, or some other factor is unknown.

37

Site Selection of Latrines

Raccoons choose specific latrine sites, often rocky, open areas (Page et al. 1998). Our results found 18 of the 40 unique latrines (45%) were on rocky locales. However, we did not determine relative availability of substrates or selection by raccoons. Latrines on our study area may have been deposited in proportion to available substrate without regard to selection — particularly because proportion of substrates at random sites was similar. The amount of coarse woody debris was significantly related to the occurrence of latrines, but unexpectedly, the distance to a large tree had no significant bearing on selection of sites. This is in contrast to Nielsen et al. (2007), who found that 61% of latrine sites were deposited at the base of large trees. Also contrary to most accounts in the literature, there was little proclivity for raccoons on our study area to form large communal latrine sites. We found only one large communal latrine at Garden of the Gods, located near a campsite, which had the only sample to test positive for B. procyonis.

Management Implications

Periodic monitoring of eastern woodrat and raccoon populations in southern Illinois should continue. Control of raccoon populations that are near reintroduction sites may be necessary should their numbers significantly increase. Raccoon populations state- wide have remained stable for the past 15 years (Bluett 2010), but as noted, they attain their highest densities in proximity to humans and/or agricultural land, where food is easy to obtain. It is in the best interest of the reintroduced woodrat populations if human impact remains at current low levels. Increased camping and recreational use may lead to increases in litter and garbage, resulting in increased raccoon abundance. The rarity of scat (and tracks) on our study area suggests a low density of raccoons and corresponding low occurrence of B . procyonis . The reintroduced woodrat populations in this area appear to be expanding, with documented reproduction and dispersal (A. K. Poole, SIUC, unpublished data), which is further indicative of low levels of B . procyonis . As such, we conclude that B . procyonis is not now a major threat to the population of woodrats in the study area.

ACKNOWLEDGEMENTS

We thank Drs. Eric Hellgren and Clay Nielsen, SIUC Cooperative Wildlife Research Laboratory, for their helpful suggestions and input to this study. Bob Bluett, Illinois Department of Natural Resources, was instrumental in efforts to reintroduce woodrats to Illinois. Several SIUC students helped locate latrine sites. We appreciate the helpful com¬ ments of two anonymous reviewers on the manuscript.

LITERATURE CITED

Barnstable, R. W. and W. G. Dyer. 1974. Gastrointestinal helminths of the raccoon, Procyon lotor, in southern Illinois. Transactions of the Illinois State Academy of Science 67:451-460.

Birch, G. L., G. A. Feldhamer, and W. G. Dyer. 1994. Helminths of the gastrointestinal tract of raccoons with management implications of Baylisascaris procyonis occurrence. Transactions of the Illinois State Academy of Science 87:165-170.

Bluett, R. 2010. 2010 spotlight survey: wildlife diversity program note #10-2. Illinois Department of Natural Resources, Office of Resource Conservation, Springfield, IL.

38

Feldhamer, G. A. and A. K. Poole. 2008. Status and conservation of other at-risk species of North American woodrats. Pages 193-206 in J. Wright and J. D. Peles, editors. The Allegheny woodrat: ecology, conservation, and management of a declining species. Springer Publishing, New York, NY.

Gehrt, S. D. 2003. Raccoon ( Procyon lotor and Allies). Pages 611-634 in G. A. Feldhamer, B. C. Thompson, and J. A. Chapman, editors. Wild mammals of North America: biology, management, and conservation. Second edition. Johns Flopkins University Press, Baltimore, MD.

Gompper, M. E. and A. N. Wright. 2005. Altered prevalence of raccoon roundworm ( Baylisascaris procyonis ) owing to manipulated contact rates of hosts. Journal of Zoology, London 266:215- 219.

Ing, D. M. 2008. The eastern woodrat ( Neotoma floridana ) in southern Illinois: assessment of pre¬ existing and reintroduced populations. M. S. Thesis, Southern Illinois University, Carbondale.

Kazacos, K. R. 2001 . Baylisascaris procyonis and related species. Pages 301-341 in W. M. Samuel, M. J. Pybus, and A. A. Kocan, editors. Parasitic diseases of wild mammals. Iowa State Univer¬ sity Press, Ames.

Kazacos, K. R., W. A. Vestre, and E. A. Kazacos. 1984. Raccoon ascarid larvae ( Baylisascaris procyonis ) as a cause of ocular larval migrans. Investigative Ophthalmology and Visual Science 25:1 177-1 183.

Kazacos, K. R. and W. M. Boyce. 1989. Baylisascaris larva migrans. Journal of the American Veterinary Medical Association 195:894-903.

LoGiudice, K. 2001. Latrine foraging strategies of two small mammals: implications for the transmission of Baylisascaris procyonis. American Midland Naturalist 146:369-378.

Mitchell, M. A., L. L. Hungerford, C. Nixon, T. Esker, J. Sullivan, R. Koerkenmeier, and J. P. Dubey. 1999. Serologic survey for selected infectious disease agents in raccoons from Illinois. Journal of Wildlife Diseases 35:347-355.

Monty, A-M., E. J. Heist, E. R. Wagle, R. E. Emerson, E. H. Nicholson, and G. A. Feldhamer. 2003. Genetic variation and population assessment of eastern woodrats in southern Illinois. Southeastern Naturalist 2:243-260.

Nielsen, C. K., A. Defore, and E. Bade. 2007. Survey of Baylisascaris procyonis and canine distemper virus in southern Illinois raccoons. Transactions of the Illinois State Academy of Sci¬ ence 100: 169-176.

Novosak, B. A. 2004. Reintroduction of the eastern woodrat (Neotoma floridana ) in southern Illi¬ nois: habitat assessment and release. M. S. Thesis, Southern Illinois University, Carbondale.

Page, L. K., R. K. Swihart, and K. R. Kazacos. 1998. Raccoon latrine structure and its potential role in transmission of Baylisascaris procyonis to vertebrates. American Midland Naturalist 140: ISO- 185.

Page, L. K., R. K. Swihart, and K. R. Kazacos. 1999. Implications of raccoon latrines in the epizootiology of Baylisascaris . Journal of Wildlife Diseases 35:474-480.

Page, L. K., S. D. Gehrt, K. K. Titcombe, and N. P. Robinson. 2005. Measuring prevalence of rac¬ coon roundworm (Baylisascaris procyonis ): a comparison of common techniques. Wildlife Soci¬ ety Bulletin 33:1406-1412.

Page, L. K., C. Anchor, E. Luy, S. Kron, G. Larson, L. Madsen, K. Kellner, and T. J. Smyser. 2009a. Backyard raccoon latrines and risk for Baylisascaris procyonis transmission to humans. Emerging Infectious Diseases 15:1530-1531.

Page, L. K., S. D. Gehrt, A. Cascione, and K. F. Kellner. 2009b. The relationship between Baylisascaris procyonis prevalence and raccoon population structure. Journal of Parasitology 95:1314-1320.

Prange, S. D., S. D. Gehrt, and E. P. Wiggers. 2003. Demographic factors contributing to high rac¬ coon densities in urban landscapes. Journal of Wildlife Management 67:324-333.

Wright, A. N. and M. E. Gompper. 2005. Altered parasite assemblages in raccoons in response to manipulated resource availability. Oecologia 144:148-156.

39

Figure 1 . Location of eastern woodrat ( Neotoma floridana ) reintroduction sites (*) in southern Illinois. The 4 sites studied for Bayliascaris procyonis prevalence were Garden of the Gods, Buzzard’s Point, High Knob, and Rim Rock. Woodrats were also introduced to Lusk Creek but that area was not included in this study. Previous extant woodrat populations (•) are at Pine Hills and Fountain Bluff.

Transactions of the Illinois State Academy of Science (2012) Volume 105, #1&2, pp. 41-49

received 9/10/11 accepted 3/28/12

Efficacy of Hoodia gordonii Extract as a Weight Loss Supplement: A Comparative Study Between an Invertebrate, Tenebrio molitor (Co I eoptera: Tenebrionidae), and a Vertebrate, Rattus norvegicus (Rodentia: Muridae)

'Justin Brohard, 'Marianne Robertson, and 2Anne Rammelsberg Millikin University, 'Department of Biology & department of Biochemistry

1 184 West Main St., Decatur, IL 62522

ABSTRACT

Despite legal constraints, Hoodia gordonii is marketed in many over-the-counter forms claiming to promote weight loss by reducing appetite. The belief that this plant is an efficacious appetite suppressant is based in traditional knowledge from the San tribes of the Namibian desert. We tested the efficacy of one commercial form of this plant extract to reduce feeding behavior, and therefore promote weight loss, in two different organ¬ isms. We used Sprague-Dawley rats, Rattus norvegicus , as a human analog to predict the usefulness of this plant as a dietary supplement for humans. We also used the adult meal¬ worm beetle, Tenebrio molitor, to compare an invertebrate species with the vertebrate results. T. molitor is a close relative of many beetle species native to the same region in which H. gordonii naturally grows. The control group contained organisms not exposed to H. gordonii. The experimental group received a body mass equivalent dosage of H. gordonii in solution with distilled water for a month. We monitored food consumption and body weight for one month to compare the control group results with those of the organisms that ingested H. gordonii solution. This study does not provide support that the commercial plant product used is efficacious as a hunger suppressing dietary supplement in either species.

INTRODUCTION

Hoodia gordonii is a desert grown plant native to South Africa and the Namib Desert that has been utilized for generations by the San people of South Africa for its supposed abil¬ ity to suppress hunger. This plant has recently been exploited and processed into a highly popular diet strategy marketed and sold in a variety of forms through multiple providers (Consumer Reports on Health, 2006). In 2003, an economical and legally binding arrangement was reached between the San tribes and the Council for Scientific and Industrial Research (CSIR) of South Africa, which allows these parties to monitor and control the production and sale of any products containing H. gordonii in an effort to pre-

42

vent over-exploitation of this resource (WHO, 2006). This treaty also gave the San peo¬ ple rights to a portion of the profit and royalties associated with the production and sale of H. gordonii-containing products (Vermeylen, 2007). With an emphasis placed on the maintenance of sufficient population levels, strict regulations have been placed on the harvesting and trade of this plant for commercial use. This limitation has often led suppli¬ ers to dilute the concentration of extract within the marketed forms (Vermaak et al., 2010).

Despite the popularity of this plant extract, little scientific data have been collected with regards to the actual effects of this plant compound. Larvae of the cabbage looper moth, Trichoplusia ni, have been used to determine the carry-over of larval H. gordonii- containing diet to the adult stage after pupation (Chow et al., 2005; Shikano and Isman 2009). Not only was this species studied to determine the effects of ingested H. gordonii, but also to determine the effects of topical application of the plant extract to the larval, pupal, and adult stages of T. ni (Akhtar et al., 2009). Ingestion of H. gordonii caused a significant change in oviposition site preference; topical application of H. gordonii had no effect on this behavior. Rader et al. (2007) determined the chemical composition of the plant extract, mainly as a method of determining the purity of marketed forms, and identified P57, an oxypregnane steroidal glycoside found in many weight loss supple¬ ments, as one of the primary components in H. gordonii.

To study the efficacy of H. gordonii on a close relative of the native South African beetle species, we chose to study the adult mealworm beetle, Tenebrio molitor. Mealworm bee¬ tles, family Tenebrionidae, are commonly referred to as darkling beetles. By testing the effects of H. gordonii extract on the feeding behavior of a close relative to the native spe¬ cies of tenebrionid beetle, we hope to develop results that can be compared to the species of beetle that would potentially feed on this plant in the wild (Hamilton et al., 2003). If H. gordonii does act as a hunger suppressant, and the tenebrionid beetles naturally exposed to this plant ingest its material, the overall food intake over the lifetime of an individual beetle may be significantly reduced. This reduction in caloric intake, and thereby reduc¬ tion in energy gain, may result in decreased reproductive success of those that consume the plant. The small size of this beetle makes it an ideal organism to study because it is cost and space efficient. This efficiency allows us to increase the sample size of our test groups, allowing the study to more accurately represent the variability present in natural settings.

We also studied the efficacy of H. gordonii on Sprague-Dawley rats, Rattus norvegicus , because of the common usage of this organism as a human analog species for pre-clinical research (Boozer and Herron, 2006). It is important to test the efficacy of pharmaceutical candidates on a human analog prior to performing clinical research, and the Sprague- Dawley rat is historically a popular choice for this type of research (Fallon et al., 2008). R. norvegicus is a relatively accessible and affordable species making it a popular choice as a research organism. Not only does research on rats allow us to predict the efficacy of H. gordonii as a weight loss supplement for humans, but it also allows us to conduct a comparative study on two organisms that are not closely related. This comparative analy¬ sis should provide information about any possible differences in the efficacy of//, gordo¬ nii as a hunger suppressant in invertebrates versus vertebrates. Although the size of these

43

rats limits the sample size, it also increases the accuracy of our techniques for administra¬ tion of H. gordonii solution.

Our hypothesis, based on the popularity of commercialization and traditional use by the San tribes of South Africa, is that ingestion of H. gordonii will suppress the appetite of both T. molitor and R. norvegicus. This suppressing effect should be represented by a decrease in average daily food consumption over the trial period and subsequent weight loss in the organisms whose diet has been supplemented with H. gordonii solution at the end of the trial period.

MATERIALS AND METHODS

We conducted this research in the Leighty-Tabor Science Center at Millikin University in Decatur, Illinois. The control group received no plant extract solution and the experi¬ mental group received a body mass equivalent dosage of H. gordonii extract in solution with distilled water. Although the concentrations of the solution used for each trial dif¬ fered based on the average starting weight of the animals, we prepared these solutions using the same H. gordonii extract, in capsule form, purchased from Mari-Mann Herb Farm in Decatur, Illinois.

We separated the beetles and rats into control and experimental groups based on relative body size by alternately placing the lightest individual of each species into a control group and the next lightest into an experimental group. This separation method ensured that the weight distribution between the two groups was almost equivalent, thereby eliminating body size as a variable on feeding rates. Although the measurement technique used for each organism differed because of the size of each organism used, we collected the same data for each trial. In order to test the efficacy of H. gordonii as an appetite suppressant, we measured the mass of food consumed. We then calculated the mean of these measurements to represent the average daily food consumption for each organism. We also recorded the body weight of each animal for the duration of each trial. At the end of the trials, we calculated the average final mass of each group of organisms and used this measurement to determine if the H. gordonii solution had caused a difference in weight. Since the average initial weights of each group were nearly equivalent, we were able to use only the final body mass to determine the efficacy of H. gordonii as a weight loss dietary supplement.

Due to the inconsistency present in the dietary supplement market, future research could address a chemical analysis of the H. gordonii extract used in this experiment. Since P57 has been identified as the active ingredient in H. gordonii, it would be useful determine the amount of compound, if any, present in the commercial extract used. High-perfor¬ mance thin layer chromatography (HPTLC) can be used to determine the presence of P57 (Vermaak et al., 2010). We used this method of biochemical analysis to determine that P57, the suspected active ingredient of H. gordonii, is present within the extract used in this research (Fig. 1). High-performance liquid chromatography (HPLC) can also be used to form a chemical fingerprint of the plant extract for comparison to the known chemical composition of pure H. gordonii (Avula et al., 2007). We are currently pursuing further methods of sample authentication.

44

T. molitor protocol

We separated 100 male and female adult T. molitor beetles into two groups, control (n = 50) and experimental (n = 50). We supplied control beetles with distilled water and experimental beetles with a constant dose of H. gordonii plant extract in solution with distilled water. We prepared water dishes by taping the bottoms of two 60.0 mm polysty¬ rene petri dishes together to form a sealed container, which we filled with two cotton balls prior to sealing. We then drilled a hole in the side of this container just big enough for the beetles to fit their heads inside to access the water supply. These containers were constructed for the purpose of controlling levels of evaporation; preliminary test results confirmed that this container design limited evaporation to an insignificant level.

We used the average weight of an adult human, 70 kg, to determine the dosage of H. gordonii per body weight ratio. We used this ratio to determine the daily dosage of H. gordonii extract to administer to the beetles based on the average mass of the beetles. We determined the total dosage for the entire group of beetles for the length of the trials and created a 1.0 L stock solution containing 0.024 g of powdered H. gordonii extract. Once we created this stock, we used a glass pipette to fill each water container in the experi¬ mental group with 1.0 mL of this solution. We filled each container in the control group with 1.0 mL of distilled water. We prepared the food for the beetles by mixing equal weights of wheat flour and whole oatmeal.

We individually housed each beetle in a 475 mL plastic container with approximately 100 mL of the prepared food mixture and covered the container with a ventilated plastic lid. We maintained each beetle in the same container for the duration of the experiment to ensure that any changes in the weights of the food were attributed to the appropriate bee¬ tle. To weigh the beetles, we removed them from their housing containers, one at a time, made sure they were free of any food particles or other debris, then placed them on their dorsal side in a plastic petri dish on the electronic balance. While the water containers were removed, and the lid was off, we weighed the entire food container. Prior to the trials, we tested the excrement production rates of the beetles by housing an individual beetle in an empty container for 24 hours. We found that, due to evaporation of waste, this production rate was insignificant when compared to the mass of the container. There¬ fore, we were able to attribute any changes in the mass of the container to consumption by the beetle being housed in that container. We tracked the weight of each container throughout the trial and calculated any change in the mass of the container that had occurred at each measurement period. We repeated this measurement process three times a week throughout the trial period of one month, recording each of the values to the near¬ est 0.001 g. We dispatched all remaining beetles by feeding them to a pet bearded dragon.

R. norvegicus protocol

We separated 22 juvenile male R. norvegicus into two separate groups, control (n = 11) and experimental (n = 11). We gavaged the control rats three times a week with distilled water and the experimental rats three times a week with H. gordonii solution (see description below and in Talpur et al., 2001). We housed rats in individual plastic containers with base dimensions of 30 x 50 x 30 cm. These containers had grated stain¬ less steel lids, equipped with food and water dispensing mechanisms, which allowed appropriate air flow and resource accessibility for the rats. We supplied the rats with ad libitum rat chow pellets. We labeled each housing container using a number between 1

45

and 22 and either an ‘A' or a lB\ The rats in the group labeled with an ‘A’ were in the control group and those with a ‘B' were in the experimental group. In order to measure the weights of the rats throughout the trial, we placed a foam bucket onto a balance to restrain the rats, eliminating the need for anesthesia, without affecting the accuracy of the measurements. The same balance was used to measure food consumption by measuring the mass of the remaining food at each measurement period and calculating the difference from the previous measurement. We weighed rats and food three times a week and rec¬ orded data to the nearest 0.01 g. At the conclusion of the trial, we dispatched all rats using an approved method of carbon dioxide euthanasia and post-euthanasia freezing.

The gavaging process is a modification of the protocol used by Talpur et al. (2001) who tested the efficacy of a mixture of Chinese herbal supplements as a weight loss supple¬ ment. We used the average weight of an adult human, 70 kg, to determine the H. gordonii dosage per body weight ratio. We used this ratio to determine the proper dosage to be administered to the rats in the experimental group based on their average initial body mass. We determined that the appropriate daily dosage, compared to 400 mg in humans, should be 12 mg per day. This dosage calculation allowed us to prepare a stock H. gordo¬ nii solution that contained 30 mg of H. gordonii plant extract per 1 mL of distilled water. This mixture, administered via a 5.08 cm curved stainless feeding needle three times a week, ensured an average dosage of 12 mg of H. gordonii per day per rat for the one month trial period.

Statistical Analysis

We calculated the mean weights for each of the groups, control and experimental, after each day of measurement, and then compared the means between the two groups using standard deviation calculations. The standard deviation of a data set is the root mean of the variance within the data set. Standard deviation utilizes the square root of the average of the deviation of each measurement from the mean; a small standard deviation indicates a data set in which most values fall close to the mean, while a large standard deviation indicates data that are spread out across the range. The standard deviation is used to cre¬ ate error bars when plotting the mean values between the two groups. If the error bars for each mean overlap one another, then there is no significant difference between the control and experimental groups. Additionally, we used a two-tailed t-test to determine the probability that any overlap was caused by random chance alone. This test determines a P- value, which is used to determine significance of any differences that may be present within the data while assuming unequal variance between the populations. A P- value greater that 0.05 indicates no significant difference in the data.

RESULTS

We omitted data collected for beetles that died before the end of the trials, leaving data for a total of 7 trial sets for 73 of the initial 100 beetles, which were able to be analyzed and compared between the two groups. The number of beetles remained close between the two groups throughout the experiment, with a total of 13 control beetles and 14 experimental beetles dying. There were no deaths in either rat groups. Both groups of T. molitor started with a mean body mass of 0.11 g. Both groups of R. norvegicus started with an initial mean body mass of approximately 210 g. This similarity in initial body mass between the control and experimental groups allowed us to use the mean final mass

46

to compare any change in body mass that might be due to ingestion of H. gordonii extract. There were no significant differences in either average final body mass or mean daily food consumption for either species. However, the average final body mass (Fig. 2) and average daily food consumption (Fig. 3) were slightly lower in the experimental group for the beetles. Similarly, the average final body mass (Fig. 4) and average daily food consumption (Fig. 5) were slightly lower in the experimental group for the rats.

DISCUSSION

Our hypothesis, that ingestion of H. gordonii plant extract would decrease food consump¬ tion resulting in a decrease in total body weight, was not supported. Although there was a trend toward decreased average daily food consumption in the experimental groups, as well as a trend toward decreased final body weight in these groups, these trends do not represent significant differences in the data. Therefore H. gordonii extract, when adminis¬ tered in a body mass equivalent dosage, does not seem to cause any significant change in feeding behavior or body weight in adult mealworm beetles or Sprague-Dawley rats.

Although H. gordonii extract did not produce significant differences in the feeding rates of T. molitor or R. norvegicus, it is still important to compare the results between the invertebrate and vertebrate subjects. In T. molitor , the difference between average daily food consumption by the experimental and control groups is larger than the difference in average final body mass of the two groups. In R. norvegicus, the difference between aver¬ age daily food consumption by the experimental and control groups is smaller than the difference in average final body mass of the two groups. Although this variation is not represented by statistically significant differences, it does shed light on a potential differ¬ ence in the type of effect, if any, which may be caused by ingestion of H. gordonii. This variation indicates that the invertebrate species might be exhibiting a behavioral response to the plant extract while the vertebrate species might be exhibiting a physiological response. The variation could also be attributed to differences in the metabolism of the two species. Although H. gordonii ingestion did not cause significant decrease in the feeding behavior of mealworm beetles, it did affect the oviposition preference of the cab¬ bage looper moth, Trichoplusia ni (Chow et al., 2005; Shikano and Isman, 2009). Inges¬ tion of H. gordonii by larval moths increased ingestion of H. gordonii in the adult moths. This indicates that H. gordonii is capable of producing an effect in an invertebrate spe¬ cies. Repeating our protocol on T. ni should allow us to further determine the efficacy of H. gordonii as an appetite suppressant in invertebrates.

We have anecdotal evidence that some of the rats, in both the control and experimental groups, exhibited behavioral changes throughout the duration of the trial. Some of the rats showed increased avoidance behavior and others showed increased aggression. Additionally, following gavaging, several rats in both groups moved quickly and ran¬ domly about the cage. Since we only have anecdotal evidence of the occurrence of these behavioral changes we cannot be sure of their cause. We can only speculate that they may be a result of associative learning.

We recognized two potential sources of error in the beetle protocol. It is possible that the large ventilation holes in the lid of the beetles’ housing containers could have affected the overall mass of the containers by allowing moisture to move freely in and out of the con-

47

tainer. Additionally, although the electronic balance used is designed for extreme preci¬ sion, other individuals were also using the same balance which could have resulted in calibration errors.

Although this research does not provide information about possible long-term effects of H. gordonii extract, it does provide some insight to the short-term effects of this plant on T. molitor and R. norvegicus. Based on our results, members of family Tenebrionidae that are naturally exposed to H. gordonii are not likely to be affected by ingestion of this plant material. Additionally, based on the lack of efficacy of this plant extract as a weight loss supplement in rats, it is likely that H. gordonii would not have a hunger suppressing effect on humans. It would be interesting to test this plant extract on humans to determine whether the San people were accurate in their predictions, but for the adult mealworm beetle and Sprague-Dawley rats, H. gordonii does not cause a significant decrease in body weight or feeding behavior.

LITERATURE CITED

Akhtar, Y., I. Shikano, and M.B. Isman. 2009. Topical application of a plant extract to different life stages of Trichoplusia ni fails to influence feeding or oviposition behavior. Entomologia Experimentalis et Applicata. 132: 275-282.

Anonymous. 2006. Hoodia: Worth trying for weight loss? Consumer Reports on Health. February:

10.

Anonymous. 2006. Protecting traditional knowledge: the San and Hoodia. Bulletin of the World Health Organization. 84:345.

Avula, B., Y.H. Wang, R.S. Pawar, YJ. Shukla, I.A. Khan. 2007. Chemical Fingerprinting of Hoo¬ dia Species and Related Genera: Chemical Analysis of Oxypregnane Glycosides Using High- Performance Liquid Chromatography with UV Detection in Hoodia gordonii. Journal of AO AC International. 90:1526-1532.

Boozer, C.N., and A.J. Herron. 2006. Simmondsin for weight loss in rats. International Journal of Obesity. 30:1143-1148.

Chow, J.K., Y. Akhtar, and M.B. Isman. 2005. The effects of larval experience with a complex plant latex on subsequent feeding and oviposition by the cabbage looper moth: Trichoplusia ni (Lepidoptera: Noctuidae). Chemoecology. 15:129-133.

Fallon, E., L. Zhong, J.K. Furne, M.D. Levitt. 2008. A mixture of extracts of black and green teas and mulberry leaf did not reduce weight gain in rats fed a high-fat diet. Alternative Medicine Review. 13:43-49.

Hamilton, W.J., J.R. Henschel, and M.K. Seely. 2003. Fog collection by Namib Desert beetles. South African Journal of Science. 99:181.

Rader, J.I., P. Delmonte, and M.W. Trucksess. 2007. Recent studies on selected botanical dietary supplement ingredients. Analytical and Bioanalytical Chemistry. 389:27-35.

Shikano, I., and M.B. Isman. 2009. A sensitive period for larval gustatory learning influences subsequent oviposition choice by the cabbage looper moth. Animal Behaviour. 77:247-251 .

Talpur, N.A., B.W. Echard, V. Manohar, H.G. Preuss. 2001. Influence of a combination of herbs on appetite suppression and weight loss in rats. Diabetes, Obesity, and Metabolism. 3:181-185.

Vermaak, I., J.H. Hamman, and A.M. Viljoen. 2010. High performance thin layer chromatography as a method to authenticate Hoodia gordonii raw material and products. South African Journal of Botany. 76:1 19-124

Vermeylen, S. 2007. Contextualizing ‘fair’ and ‘equitable’: The San’s reflections on the Hoodia benefit-sharing agreement. Local Environment. 12:423-436.

Figure 1. Comparison of HPLC profiles of P57 standard solution (A), H. gordonii extracted with acetonitrile (b), and H. gordonii extracted with acetonitrile and water (50:50), at a detection wavelength of 220 nm.

48

Is* : po - m r-~ , o> co iS) r>» o>

CO ICO iCO CO © O) Ol © CD Ol

Cwporm* fMtttfton A tea Htssjjhi unit*

Figure 2. Comparison of average final body mass (± SD) of adult Teneb- Figure 3. Comparison of mean daily food consumption by control adult

rio molitor beetles in the control group receiving distilled Tenebrio molitor receiving distilled water versus an experi-

water versus an experimental group receiving 0.024g of Hoo- mental group receiving 0.024g of Hoodia gordoniifL of dis-

dia gordonii/L of distilled water (P = 0.141). tilled water (P = 0.405).

49

(os X ueiut)

(81 pooi pnumsuca jo ssej\

.c si

£ £

3 XI

e -o fl S, hi 43 60

E t '

' i . 3

O 60

SO

(T)

3

>

3

60

Q -S C/3 -~

0C

+1 ^

co

co

03

C ^

s

>>

X w

8 OS

cx

3 O

t-

& an

X> 3

h— i

c <u

E

g c

C 3

^ O 43 U

43

a.

x

43

C

03

0)

3

£

X

43

<4-1

o

60

03 „

t- jO 0)

> C 3 O

C/5

>> 3

CO

Ch

43

>

S-i

43

3

£

X3

43

C/5

■3

e

1

o

60

a • *»•»

"Xl

o

£

Ct_

o

60

E

cn S

in

<u

u.

3

60

"4*?, * -• j

Pf®

(8) pooj poiunsuo ) jo s'vrj^

(OS * uesui)

(8) tsep* <pofi |bib;(

O

c.

4—1

c

o

o

<4-1

o

c

03

C/5

3

C/5

l-i

43

>

_ *“*

Q «

C/3 03

+, * w XI

c/3 <L>

C/3

cd ra

s i

>4 ^

o

f 1

03

C X IS <L> ^ 60

o

60

a • »««*

"XI

o

a

60

E

o

cn

(U

60

3

Ch

43

>

3

c

O

C/5

43 60

_ o3

3 >

> 3

3 60

60

CX

05 3

3 O

•3 oo 60

X —H

£ I

§ I

Oh §

E ^ 5 v- o <3

U 05

43

Ch

3

60

£

(as =f uraiu) <s) ssc(s xpoa

Control Experimental Control Experimental

50

Transactions of the Illinois State Academy of Science (2012) Volume 105, #1&2, pp. 51-55

received 1 1/7/1 1 accepted 4/22/12

Mass Death of Wintering American Robins ( Turdus migratorius) in Decatur, Illinois

Max S. Huschenand David Joseph Horn Department of Biology, Millikin University 1184 West Main Street, Decatur, Illinois 62522

ABSTRACT

A flock of American Robins ( Turdus migratorius ) congregated around a fruiting tree and nearby building on the campus of Millikin University in Decatur, Illinois in early 2011. Thirty-four robins were found dead around campus buildings during 8-17 February, after a blizzard passed through central Illinois. We investigated potential causes for the high number of deaths including examinations of gizzard contents and feces for both food eaten and internal parasites, and observed robin behavior. The robins apparently congre¬ gated around one of the few food resources available in the area during this blizzard. They were subject to starvation intensified by poor nutritional quality, parasitic infection, and aggressive defense by conspecifics, and were more susceptible to other forms of mortality including bird-window collisions.

INTRODUCTION

Mass deaths of birds are not uncommon, but largely go unnoticed. Mass deaths can be regional or local in scope and are a result of both human and natural causes including collisions with human structures (Klem 1990), poisoning (Dolbeer et al. 1995), and dis¬ ease (LaDeau et al. 2007). Poor weather is also a factor causing mass deaths (Jehl 1998). For example, while rare, starvation deaths during the winter can occur during extreme winter storms (Bednekoff and Houston 1994). We investigated the mass death of Ameri¬ can Robins ( Turdus migratorius) that had congregated around a single fruiting crabapple tree ( Malus spp.) at a building on the Millikin University campus in Decatur, Illinois. We hypothesized that starvation was the primary cause of robin mortality, and secondary factors including nutritional quality, parasitism, and aggressive defense by conspecifics intensified starvation.

METHODS

We observed American Robins at Millikin University in Decatur, Illinois starting on 8 February 201 1 when a blizzard occurred in central Illinois with temperatures reaching a low of -12°C and snow accumulation of 30.5 cm. We began receiving notifications of dead robins along the south side of Staley Library, as well as surrounding buildings, immediately preceding the blizzard and the days after. We collected dead robins and

52

observed the flock of several dozen robins that continued to congregate around the crabapple tree next to the library until the snow receded on 17 February.

Dead robins were weighed, gizzard and intestine dissected, and feces analyzed. Identification of internal parasites was performed following McQuistion and Wilson (1989). The nutritional content of the crabapples was analyzed by Eurofins Nutrition Analysis Center (Des Moines, IA, USA; % carbohydrates determined via CFR 21- calculation, % lipids calculated via acid hydrolysis - AOAC 954.02, % protein calculated via combustion - AOAC 990.03, and calories determined via CFR Atwater calculation). Ten-min observations were made of the behavior of the robins twice each day. Feeding behavior, resting behavior, and social interactions including aggressive defense of the food resource among robins were noted. We defined any instance where one robin would chase another robin from the food resource as aggressive defense (Pietz and Pietz 1987).

RESULTS

Thirty-four dead robins were found over the 10-day period. Twenty-six robins were weighed, gizzard and intestine contents analyzed, and feces examined for internal para¬ sites. The other eight robins had been scavenged or decomposed prior to collection. The mean weight of 26 robins was 50 g (range from 37 to 67 g). Eighteen of 26 robins (69%) had empty gizzards. Four of eight robins with full gizzards had whole undigested crabap¬ ples within the gizzard. The other four robins had unidentifiable food in the gizzard. The crabapples contained 30.9% carbohydrates, 1.2% lipids, 2.5% protein, and an energy con¬ tent of 1.45 kcal/g. Thirteen of 26 robins (50%) had internal parasites. Six robins had parasites of the phylum Acanthocephala. We found parasites of the genus Capillaria in five robins and in one robin we found Hymenolepis (spp.). Three of 26 robins were in a position directly underneath windows where window collisions have been observed in the past (DJH, unpubl. data). We noted at least three instances of aggressive defense of the food resource during our behavioral observations.

DISCUSSION

On average, American Robins weighed 77 g throughout the year in Pennsylvania and 86 and 84 g for males and females, respectively, during winter in Ithaca, New York (Sal- labanks and James 1999). The weight of every dead robin we found was at least 10% under the average weight of 77 g recorded for robins in Pennsylvania. Many species, particularly smaller birds, maintain fat reserves below their physiological capacity in win¬ ter and have only enough reserves to survive the coldest night at a certain location at a certain time (Lima 1986, Krams et al. 2010). Thus, American Robins at our location may not have had the fat reserves necessary to survive multiple days in below-zero tempera¬ tures with limited access to food. This may be a result of a trade-off between the benefits of fat reserves to prevent starvation and the risk of predation (Lima 1986). Birds with higher fat reserves may be more likely to survive cold winter nights, but they are also more vulnerable to predation due to increased body weight and decreased take-off speed (Lima 1986). We conclude the mass kill was primarily a result of starvation. Other fac¬ tors possibly contributed including the nutritional quality of the food source, parasitism, aggressive defense, and collisions with windows.

53

Nutritional Quality

The robins' main food source was apparently limited to crabapples because of the bliz¬ zard, and particularly the ice and snow on the ground. These fruits may not have met their nutritional requirements. Studies have found robins will consume more fruits with higher lipid concentrations during the autumn (Lepczyk et al. 2000). Lipid-rich fruits may be important for fattening, as thrushes achieve the highest energy assimilation rates when choosing fruits high in lipid content (Witmer and Van Soest 1998, Lepczyk et al. 2000). At a time when energetic demands were high in the robins we studied, the amount of lipid-rich fruits in their diet and presumably energy assimilation rates was low.

A 55-g robin requires 30.7 kcal/day (Hazelton et al. 1984). Thus, a similar-sized robin in our study would have needed to eat 21 g, or 74 crabapples, per day to meet nutritional requirements. Crabapple seeds also contain cyanide (Seigler 1976), and this poison can cause death when consumed in large amounts by Cedar Waxwings ( Bombycilla cedrorum, Woldemeskel and Styer 2010). We found evidence in the gizzards of partially digested crabapple seeds and it is possible these robins may have been exposed to cyanide.

Parasitism

American Robins are known to be infected by a large number of parasites including coc- cidia, trematodes, nematodes, cestodes, and acanthocephalans (Welte and Kirkpatrick 1986). Collectively, over 40 helminth parasites have been identified in robins (Cooper and Crites 1976). In our study, parasites were found in 50% of the dead robins, and three different types of parasites were discovered. Robins with parasitic infections may have reduced metabolism, inactivity, and weight loss at the time where increased metabolism and activity were most needed. Acanthocephalans are known to reduce metabolic rate in European Starlings ( Sturnus vulgaris , Atkinson et al. 2008). Capillaria spp. are digestive tract parasites that have been found to cause inactivity, anorexia, weight loss, vomiting, and death in guinea fowl (De Rosa and Shivaprasad 1997). Rates of parasitic infection in other studies were similar to ours and ranged from 33-77% (Welte and Kirkpatrick 1986, McQuistion and Holmes 1988).

Aggressive Defense

We observed multiple instances of defense of the food source. Typically, one robin would chase other robins to gain access to fruits on the ground. In robins, defense of fruits is rare except when temperatures are extremely low in which case it will last for a few days (Sallabanks 1993). Robins with winter territories have feeding bouts that are five times longer, ingest twice as much food per bout, but forage half as fast as robins that intrude onto a territory (Sallabanks 1993). Given the large food requirements and defense displayed, some robins may not have had access to the food source and subse¬ quently did not meet their nutritional requirements.

Window Collisions

The number of bird-window collisions is approximately 7-8 birds per building per year on the Millikin campus, and the library is one of the buildings with the most collisions (DJH, unpubl. data). The proximity of the crabapple tree to the building may have influ¬ enced the number of window collisions. Birds can become intoxicated from eating large amounts of overripe fruit (Woldemeskel and Styer 2010), which may in turn make them more vulnerable to collisions.

54

There were several factors that we were unable to control in this study. Given the cold weather, we were not able to capture living birds in the area and weigh them to compare the weights of living birds with the dead ones we found. For the same reason, we could not compare parasitism rates between surviving birds with those that died. Finally, future studies should compare the number of birds that died to the number of living birds in the area, and perform additional behavioral observations.

Singularly, factors such as starvation, low nutritional quality, cyanide poisoning, parasit¬ ism, resource defense, and window collisions can result in bird death. Collectively, we suggest that these factors, and particularly starvation, resulted in a kill of 34 American Robins under harsh and stressful weather conditions at Millikin University in February 2011. While the death of 34 robins is small relative to the overall population of robins in central Illinois, this study can provide insight into the causes of future localized mass deaths.

ACKNOWLEDGMENTS

We thank T. E. McQuistion and T. E. Wilcoxen for assistance in this study and for

reviewing earlier drafts of the manuscript. A. M. Johansen, L. A. Lundstrom, and two

anonymous reviewers also reviewed the manuscript prior to submission. Funding was

provided by the Millikin University Fund for Ornithological Research.

LITERATURE CITED

Atkinson, C. T., N. J. Thomas, and D. B. Hunter. 2008. Parasitic diseases of wild birds. Wiley- Blackwell, Ames, Iowa, USA.

Bednekoff, P. A. and A. I. Houston. 1994. Optimizing fat reserves over the entire winter: a dynamic model. Oikos 71:408-415.

Cooper, C. L. and J. L. Crites. 1976. A check list of the helminth parasites of the robin, Turdus migratorius Ridgway. American Midland Naturalist 95:194-198.

De Rosa, M. and H. L. Shivaprasad. 1997. Capillariasis in Vulture Guinea Fowl. American Association of Avian Pathologists 43:131-135.

Dolbeer, R. A., D. F. Mott, and J. L. Belant. 1995. Blackbirds and starlings killed at winter roosts from PA-14 applications, 1974-1992: implications for regional population management. Proceedings of the Eastern Wildlife Damage Management Conference 7:77-86.

Hazelton, P. K., R. J. Robel, and A. J. Dayton. 1984. Preferences and influence of paired food items on energy intake of American Robins and Gray Catbirds. Journal of Wildlife Management 48:198-202.

Jehl, J. 1998. Conspecific collisions can precipitate mortality in migrating Eared Grebes. Wilson Bulletin 110:409-411.

Klem Jr., D. 1990. Collisions between birds and windows: mortality and prevention. Journal of Field Ornithology 61:1 20- 1 28 .

Krams, I., D. Cirule, V. Suraka, T. Krama, M. J. Rantala, and G. Ramey. 2010. Fattening strategies of wintering Great Tits support the optimal body mass hypothesis under conditions of extremely low ambient temperature. Functional Ecology 24:172-177.

LaDeau, S. L., A. M. Kilpatrick, and P. P. Marra. 2007. West Nile virus emergence and large-scale declines of North American bird populations. Nature 447:710-714.

Lepczyk, C. L., K. G. Murray, K. Winnet-Murray, P. Bartell, E. Geyer, and T. Work. 2000. Sea¬ sonal fruit preferences for lipids and sugars by American Robins. Auk 1 17:709-717.

Lima, S. L. 1986. Predation risk and unpredictable feeding conditions: determinants of body mass in birds. Ecology 67:377-385.

55

McQuistion, T. E. and B. B. Holmes. 1988. Isospora robini sp. n., a new coccidian parasite (Apicomplexa: Eimeriidae) from the American Robin ( Turdus migratorius) . Proceedings of the Helminthological Society of Washington 55:324-325.

McQuistion, T. E. and M. Wilson. 1989. Isospora geospizae, a new coccidian parasite (Apicom¬ plexa: Eimeriidae) from the Small Ground Finch ( Geospiza fuliginosa ) and the Medium Ground Finch ( Geospiza fortis ) from the Galapagos Islands. Systematic Parasitology 14:141-144.

Pietz, M. A. J. and P. J. Pietz. 1987. American Robin defends fruit resource against Cedar Wax- wings. Journal of Field Ornithology 58:442-444.

Sallabanks, R. 1993. Fruit defenders vs. fruit thieves: winter foraging behavior in American Robins. Journal of Field Ornithology 64:42-48.

Sallabanks, R. and F. C. James. 1999. American Robin ( Turdus migratorius). The Birds of North America. Number 462.

Seigler, D. S. 1976. Plants of the northeastern United States that produce cyanogenic compounds. Economic Botany 30:395-407.

Welte, S. C. and C. E. Kirkpatrick. 1986. Syngamiasis in juvenile American Robins ( Turdus migratorius ), with a note on the prevalence of other fecal parasites. Avian Diseases 30:736-739.

Witmer, M. C. and P. J. Van Soest. 1998. Contrasting digestive strategies of fruit eating birds. Functional Ecology 12:728-741.

Woldemeskel, M. and E. L. Styer. 2010. Feeding behavior-related toxicity due to Nandina domes- tica in Cedar Waxwings (Bombycilla cedrorum). Veterinary Medicine International 2010:2042- 2048.

Transactions of the Illinois State Academy of Science (2012) Volume 105, #1&2, pp. 57-64

received 3/21/1 1 accepted 1/30/12

Conservation Review of the Longnose Dace Rhinichthys cataractae (Valenciennes) in Northwestern Illinois

Jeremy S. Tiemann1*, Christopher A. Taylor1, and Jason Knouft2 ‘Illinois Natural History Survey, Prairie Research Institute, University of Illinois

1816 South Oak Street, Champaign, IL 61820 2St. Louis University, Department of Biology 3507 Laclede Avenue, St. Louis, MO 63103 Correspondence: jtiemann @ illinois.edu

ABSTRACT

The longnose dace Rhinichthys cataractae (Valenciennes) is a small, elongated, slightly dorsoventrally compressed minnow that possesses the widest distribution of any North American cyprinid. In Illinois, it is considered rare and currently is known from streams in the Wisconsin Driftless Area and the shoreline of Lake Michigan and some of its tributaries. We examined the distribution, population status, and habitat requirements of R. cataractae in the Wisconsin Driftless Area in Illinois. The dace was collected at 12 of 33 sites sampled, and catch-per-unit-effort (number of individuals per hour collecting) varied from 0.8-52 per site. It was found most often in gravel/cobble riffles in small- to medium-sized streams. Although its range is limited in Illinois, the dace is locally abun¬ dant in several basins and we feel it does not warrant listing at this time.

Key Words:

Longnose dace, Rhinichthys cataractae , Wisconsin Driftless area, endangered species

INTRODUCTION

The longnose dace Rhinichthys cataractae (Valenciennes) is a slightly dorsoventrally compressed minnow (Cyprinidae) with a long fleshy snout. The species can reach 15 cm in length (Smith, 1979; Becker, 1983) and possesses the widest distribution of any North American minnow, generally occurring in areas above 40°N (Page and Burr, 1991). Rhinichthys cataractae is distributed from north of the Arctic Circle south to the Appala¬ chian Mountains and west to the Rocky Mountains. Individuals occupy gravel/cobble riffles in small- to medium-sized cool-water streams and in wave swept shallows of the Great Lakes. In Illinois, the species is considered rare (Smith, 1979) and has been found only in a few of the streams of the Wisconsin Driftless Area (e.g., Menominee, Little Menominee, Sinsinawa, and Plum river basins) in Jo Daviess and Carroll counties, along the shores of Lake Michigan and in a few streams that feed into the Lake in Cook and Lake counties. Records also exist from Union County but the dace is considered extir¬ pated from southern Illinois (Smith, 1979).

58

While R. cataractae has been a known component of the Illinois fish fauna since 1884 (Forbes, 1884), little is known about its range and habitat preferences in the Wisconsin Driftless Area of the state. The species was reported to be “very rare” in inland waters by both Forbes and Richardson (1920) and Smith (1979). To address these uncertainties, we conducted the first targeted status survey of R. cataractae in northwestern Illinois. The objectives of our study were to 1) gather distribution and abundance data for the Illinois Endangered Species Protection Board to assist in listing decisions under the Illinois Endangered Species Act; 2) provide valuable natural history data (e.g., habitat require¬ ments) on one of Illinois’ rarest fishes; and 3) use geographic information systems to determine if geologic, hydrologic, or landscape variables can predict the distribution of R. cataractae. Such data can assist natural resource agencies in determining geographic regions that may hold the highest potential for long-term protection of the longnose dace.

METHODOLOGY

Fish sampling - Thirty-three sites were sampled in streams within the Wisconsin Drift¬ less Area (Table 1; Fig. 1) from 18 June 2008 - 18 June 2009. Sites were selected based on either historical records for R. cataractae or habitat characteristics (e.g., gravel/cobble riffles in small- to medium-sized cool- water streams) that looked suitable for the dace. Historical records were defined as collection locations from unpublished literature (e.g., internal reports from Illinois Department of Natural Resources, Western Illinois Univer¬ sity, or U.S. Fish and Wildlife Service) or locations for which vouchered specimens exist at a museum. Of the 12 fish collections contacted, only three (Field Museum of Natural History [FMNH], Chicago; Illinois Natural History Survey [INHS] Fish Collection, Champaign; Milwaukee Public Museum [MPM], Milwaukee) had records from our study area. At most sites, fishes were collected for 45 minutes using a barge electro-shocker set at 200 volts; however, a few sites were too small to accommodate the barge and therefore were sampled using a 3.05 m minnow seine for 45 minutes. Fishes were identified, counted, and released upon completion of sampling at a site, and at least one dace was vouchered from each site and deposited in the INHS Fish Collection. Four historical sites (Table 2) were not visited because access could not be gained or R. cataractae had been collected at the site within the past five years.

Geographic Information Systems - The potential distribution of R. cataractae was pre¬ dicted using georeferenced locality data, GIS environmental layers, and the Maxent spe¬ cies distribution algorithm (Phillips et al., 2006). Maxent is a general-purpose machine learning approach to modeling of species distributions using presence-only data (Phillips et al., 2006). Maxent predicts potential distribution of a species by estimating probability distribution of maximum entropy across a specified region, subject to a set of constraints that represent incomplete information about the target distribution (Phillips et al., 2006). Locality data across the range of R. cataractae in Illinois was integrated with landcover and physical GIS data to predict potential areas of occurrence in the state. GIS data included measures of elevation, slope, flow accumulation, drift thickness, and landcover characteristics (e.g., riparian forest density). The predicted distribution of R. cataractae was then compared to sites not containing the species to further explore the importance of abiotic and biotic factors regulating the distribution and presence of R. cataractae.

59

RESULTS

Distribution and population status - Rhinichthys cataractae was collected at 12 of the 33 sites sampled, and catch-per-unit-effort (standardized by number of individuals collected per hour of collecting effort) varied from 0.8-52 per site (Table 1). The species was col¬ lected throughout the Menominee, Little Menominee, Sinsinawa, and Plum river basins and in two sites in the Galena River basin (Fig. 1). The species was most often collected in areas with swift flows over gravel/cobble riffles with depths ranging from 0.1 -0.5 m. Even though there are unconfirmed records available (e.g., internal reports from Illinois Department of Natural Resources, Western Illinois University, or U.S. Fish and Wildlife Service), we failed to collect R. cataractae in the Apple or Rock river basins and voucher specimens do not exist for this species in these basins. We believe the dace does not occur in the Apple or Rock river basins and all records have been based on misidentified blacknose dace Rhinichthys atratulus. However, we cannot rule out that R. cataractae might have possibly become extirpated in these basins.

Geographic Information Systems - Fish localities were evenly divided for development and testing during Maxent species distribution model generation. The Maxent model pro¬ duced a significant prediction of the distribution of R. cataractae across the study area ( AUC = 0.742, P = 0.041). Landcover classification (66.3%) and flow accumulation (33.6%) contributed 99.9% of the explanatory power of the model prediction, suggesting these two variables are of primary importance in predicting presence of the species. Finally, the distribution model generated for R. cataractae was tested using sites where no dace were collected. In this case, the Maxent model was not able to predict areas with¬ out R. cataractae (AUC = 0.635, P = 0.092). This result suggests sites where the fish was absent in our collections do not contain suitable habitat for R. cataractae based on the GIS data sets used to generate the models.

DISCUSSION

Rhinichthys cataractae appears stable in Illinois. Our field survey found the species at a total of 12 sites (Table 1). Of those sites, four were ones at which the species had been previously collected and eight were new. Of the eight new sites, two were in the Galena River drainage. These two records represent the first vouchered records for the species in this drainage in Illinois. While population sizes in the Galena River drainage sites were small (<1 .5 individuals per hour), habitat at those sites was identical to that found at other locations with larger populations. Further work will be needed in the Galena River drain¬ age to determine if significant population changes are occurring.

Habitat at sites containing R. cataractae was relatively uniform. The species was always found in areas with moderate to strong flow and substrates of clean, cobble sized (0.1 -0.3 m diameter) rocks. There was variation in stream width (8-20 m) and water depth (0.1- 0.5 m) among sites.

The results of the Maxent species distribution models suggest presence of R. cataractae can be predicted by landscape-level variables, particularly flow accumulation (essentially a measure of how far downstream the species occurs) and riparian landcover type. The dace primarily was found at sites with upland forest (41.7% of sites) and rural grassland

60

(25.0% of sites) riparian landcover classifications and in more upstream locations within drainage basins. These habitat characteristics are consistent with those reported for the species from across its range (Smith, 1979; Becker, 1983; Aadland, 1993; Goldstein, 2009).

With the exception of the two Galena River drainage populations discussed above and one Carroll Creek site located in Mt. Carroll, longnose dace were relatively common in suitable habitat at most sampling sites (Table 1). Number of individuals collected per hour of sampling effort ranged from 0.8-52.0. During our survey, we not only docu¬ mented the species in a new basin, but also showed the dace continues to occur at multi¬ ple sites in the Menominee, Little Menominee, Sinsinawa, and Plum river drainages. Also, R. cataractae occurs at several sites in the Wisconsin side of these basins (data from MPM). When combining these factors, we feel the longnose dace does not warrant listing at the state level as threatened or endangered at this time. We did not assess the status of Lake Michigan populations in Illinois. However, the fish has been recently (post- 1980) collected throughout the area including the Lake and some of its tributaries in Cook and Lake counties, Illinois, Lake County, Indiana, and Kenosha County, Wisconsin (Retzer and Batten, 2005; data from FMNH, INHS, and MPM).

The longnose dace is considered a cool-water, sensitive species (Lyons et al., 2010). The fish’s preference for clean cobble substrates in northwestern Illinois streams highlights one potential threat for the species. Lyons et al. (2010) predicted climate warming could affect the distribution of R. cataractae. Also, non-point source pollution by siltation can overlay cobble and prevent fish from taking shelter and/or feeding in interstitial spaces. While agricultural and livestock activities have been prevalent in northwestern Illinois for 100-150 years and some degradation of streams in that region has occurred, continued efforts must be made to limit suspended solid input into streams to protect populations of longnose dace and other aquatic taxa.

ACKNOWLEDGMENTS

The project was partially funded by the Illinois Department of Natural Resources’ Illinois Endangered Species Protection Board (#RC08E07W) and by the Illinois Department of Transportation. The Wisconsin Department of Natural Resources issued a Wisconsin collecting permit. B. Cheek, D. Ellington, J. Sherwood, M. Ulrich, and M. Wilson assisted with fieldwork. F. Veraldi shared records from the Lake Michigan area, and P. Willink (FMNH) and M. Pauers (MPM) generously provided access to specimens under their care.

LITERATURE CITED

Aadland, L.P. 1993. Stream habitat types: their fish assemblages and relationship to flow. North American Journal of Fisheries Management 13:790-806.

Becker, G.C. 1983. Fishes of Wisconsin. University of Wisconsin Press, Madison. 1052 p.

Forbes, S. A. 1884. A catalogue of the native fishes of Illinois. Report of the Illinois State Fish Commissioner for 1884:60-89.

Forbes, S. A. and R. E. Richardson. 1920. The fishes of Illinois. 2nd Edition. Illinois Natural History Survey. 357 p.

61

Goldstein, R.M. 2009. Interpreting stream physical characteristics Index of Biotic Integrity classifications: general similarities and specific differences. North American Journal of Fisheries Management 29:151-162.

Lyons, J., J.S. Stewart, and M. Mitro. 2010. Predicted effects of climate warming on the distribu¬ tion of 50 stream fishes in Wisconsin, U.S.A., Journal of Fish Biology 77: 1867-1898.

Phillips, S.J., R.P. Anderson, and R.E. Schapire. 2006. Maximum entropy modeling of species geographic distributions. Ecological Modelling 190:231-259.

Retzer, M.E. and B. Batten. 2005. Fishes of the Chicago region: A review of the Dennison and Illinois Natural History Survey Collections. Transactions of the Illinois State Academy of Sci¬ ence 98:63-73.

Smith, P.W. 1979. The Fishes of Illinois. University of Illinois Press, Urbana. 314 p.

Table 1. Sites sampled during the 2008-2009 Rhinichthys survey. Dace column indicates which Rhinichthys species were present at a given location and catch-per-unit-effort is standardized by number of R. cataractae per hour collecting. Asterisk (*) indicates that two individuals were vouchered but CPUE was not determined.

62

w

d

a.

CJ

u

o

Cd

a

OD|

a

o

03

c

o

cd

o

_o

c

o

£

E

o

O

c

3

o

U

d

cd

c/3

E

cd

D

—

GO

ooh

of oo ^ *

©

of

oo ro

o Z

no

n 6

O O O CO

o

or

(N CN O OO CN

4i

Q

5

<3

CJ

d)

d3

CJ

CJ

43

CJ

CJ

CJ

43

Q CJ Q

a 5

C3

3 CJ 5

3 CJ 3

3 CJ 3

«>3

co S

»3 s

co S a

Co a a

3 CJ 3

Co "a

co a a

Cd 444 CJ Cd v

Cd 3

CS ■^k a Jk.

Co a "*-^k a

Co a ^k a

cc

Cl

Cd

Cd

c:

Cd

C3

C3

Cd

Cd

Cd

C3

a

C3

Cd

Cd

cs

cs

Cd

cs

C3

C3

cs

>-«* Cd

!<

C5

Cd

Cd

cs

•v^k

■^k

CJ

O

CJ

CJ

CJ

CJ

Cd

Cd

Cd

Cd

C3

Cd

CJ

C3

Q

CJ

CJ

CJ

Cd

cs

0^

oq

oq

oq

oq

Qd

Qd

Qc;

oq

o

<N

ON

o

r-

o

or

or

On

or

ON

oo

4—

o

©

©

or

DO

Os

so

SO

r-

DO

©

ON

Os

r-

oo

ro

or

©

or

oo

CO

OO

00

On

CN

oo

r"

oo

oo

OO

NO

OO

r-

oo

DO

CN

Os

so

r-

CN

CN

ro

so

or

CN

1—1

r-

r^

r-

or

4— <

CN

NO

00

CO

OO

NO

r-

CN

oo

CN

ON

o

CN

©

DO

©

OO

©

or

OO

r-

r-

ro

oo

©

or

OO

so

so

CN

DO

so

On

CO

N"

oo

ON

oo

o

r-

00

ON

O

ON

NO

oo

©

DO

4—1

oo

©

or

DO

DO

Os

r-

ro

r-

©

or

r-~

DO

CN

so

oo

oo

oo

or

'Cf

or

or

cq

rq

cq

or

rq

rq

ro

ro

©

>— 1

rq

i— <

CN

CN

©

i— j

©

On

NO

ON

Os

©

4*2

©

©

o

o

o

o

O

o

O

o

o

o

o

O

©

©

©

©

©

©

©

©

©

©

©>

©

©3

ON

ON

ON

ON

©

©

©

©

ON

On

ON

ON

On

ON

ON

ON

ON

ON

ON

ON

On

©

ON

©

©

os

©

Os

Os

Os

Os

Os

OS

OO

00

OO

oo

ON

Os

Os

Os

£

£

&

£

&

£

£

&

£ £

no"

NO"

c-~"

CN

O

r-

NO

CO

OO

or

or

ro

or

or

Os"

or

NO

oo"

SO

SO

ro

Os

ro

o r

or

— T

©"

t"-"

©"

do"

ro

or

oo

NO

oo

C^

r~

NO

CN

o

NO

ro

OO

oo

oo

so

ro

ro

o r

Os

CN

oo

oo

Os

Os

Os

r-

r-

CN

so

Os

^r

r-

ON

NO

CN

NO

co

or

(N

NO

—4

oo

ro

00

or

ro

ro

©

©

Os

CN

©

CN

©

SO

or

©

oo

DO

ro

or

o

oo

NO

r-

OO

CO

r-

oo

OO

o

ro

On

c-

ro

or

©

DO

.—I

SO

ro

CN

i — i

oo

DO

r-

ro

Os

©

©

or

DO

CN

oq

'cT

■'CT

or

or

or

cq

rq

rq

rq

CN

CN

CN

or

or

rq

rq

rq

rq

ro

rq

CN

CN

CN

©

«— j

»— 2

©

©

<N

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

CN

^r

'Ct

Nt

'Ct

or

or

or

nT-

or

or

oT

or

or

or

or

or

or

or

or

or

or

or

or

or

or

or

or

or

or

or

z

Z

Z

z

Z

z

Z

Z

z

z

Z

Z

Z

Z

Z

Z

Z

Z

Z

Z

Z

Z Z

z z z z

Z

Z

Z

Z Z

i-J i-J i—3 ►— 3 hJ i-J i—3 i—3 i—3 i-4 i-J i—3 ►— 3 ►— 3 >— 3 i— 1 — 3 i-J —3 —3 — 3 «— 3 — 1 i — 3 i_ 3 —3 — 1

£	E	E	E	E	E
	oo	oo	CN	m	CN
ro	NO	NO	DO	of	cn
C/3	C/3	C/3	C/3	C/3	C/3
C/3	C^	C/3	C/3	C/3	C/3
D	D	D	D	D	D
>	>	>	>	>	>
03	c^3	03	03	03	03
Q	Q	Q	Q	Q	Q
o	o	o	o	o	o
	*—)				5
X	_3	kJ	X	J	U
				H- H
		Uh
	D	D
	>	>
	CC	oi
	a/	D
CL)	D	D	u-	l-	U.
>	C	C	D	D	D
2	"S	c	>	>	>
D	G O	G O	od	2
D	c	c	03	03	03
— 'g	D s	D s	£ 03	03	03
o	D	<L>	_c	_c	_c
c D	-4—*	-4—* -4— »	C/3 G	C/3 G	C/3 G
s		2	GO	GO	33

— < co © cn r-~ oo co

E E £ E

CN Os CO © of of CN CN

E £ E

no co r-~ do co

^ i-q on co cn of ro do do

C/3

C/3

d

’>

cd

a

1/3

C/3

o

">

cd

Q

C/3

C/1

O

'>

cd

Q

o o o

C/3

C/3

a/

’>

cd

a

o

C/3

C/3

O

’>

cd

Q

o

1/3

C/3

-

'>

cd

Q

C/3

C/3

—

>

cd

Q

o o

c/3

C/3

a/

’>

cd

Q

o

C/3

C/3

o

’>

cd

a

o

C/3

C/3

D

'>

cd

Q

C/3

C/3

O

’>

cd

Q

C/3

C/3

a/

■>

cd

Q

1/3

1/3

a/

>

cd

Q

o o o o

C/3

C/3

a/

’>

cd

Q

o

C/3

C/3

a/

'>

cd

Q

o

C/3

C/3

-

’>

cd

a

C/3

C/3

O

■>

cd

Q

C/3

C/3

—

>

cd

Q

o o o

C/3

C/3

.o

>

cd

Q

o

e

o

1/3

c

d © P3 o ex t « e3

O

fc

cd

O

t

cd

O

t

cd

O

t

cd

o

fc

cd

-^f^U&oUUUUUU

a/

>

2

cd

e

_d

cd

a

J*d

c-

O

ti¬

ki

<U

>

S

cd C

0) *4

—

a w

o

Ui

^ u

D

V-

u

o

Ou

X

o cx ,

cd

E

_ C/3

cd o

E -c

GO H

c- •— a/ a/ > _> 2 2

'S/B.

c/3 *c/3 C/3 (X)

5 CO

C/3 C/3

is is

rS rS C 'C H H

d

M -X >

<U <u • o

<u o a

CJ U

£

o

a.

CX

So

C/3

o

3C

o

K

o

cd cd D U 'C CQCQh

0)

>

2

_a>

a.

Q.

<

CD

>

M

CD

<D

CJ

£

o

ID

-X

D

D

C* o

HJ d) I 'o.'a. -c CX CX.22 < < ■

5t

_o

X

4=!

C/3

£

Jkd			D s_			u
D	D		rj			D >	-X	-X
D N.	D Nk	-X	>— ✓ r~|	t-i		D D	D D	D D
u D o	u D O	D D li u	c^ G	D > 2	D > '2	o — a	L* u	u	l_ u
cd E	cd E	— C/3	-4—*	E	E	D C G	"o t	"o t	"o fc
3	3	3	•4—*	G	3		3	3	3
PJ	Ph	ai	2	cu	Oh	u	u	u	u

-X

D

-X D D -X

D D CJ

s- CD

^CJ §

O. , C/3

E d E

Cd d ^

U Q £

Table 2. Historical sites where Rhinichthyes cataractae has been collected but were not sampled because sites were either inaccessible (I) or a recent (R) record exists for the fish since 2005. Specimens housed at the Illinois Natural History Survey Fish Collec¬ tion, Champaign.

63

c

o

</3

co

<D

OC

"O

>

tH

<u

</)

X>

o

•4— •

C/5

J

00

c

o

CO

d

4—*

c

3

O

U

oi

4—*

CO

4—*

C/3

s

C3

<l)

■— •*— >

on

Di •— 1 *— 1

£ on cn £ O os vo ^

8 ! >ȣ

3 d 3 3

•— > <« H-, I— ,

(N X h fCl CM (N -h <N

on On cm m NO Tt

>n in o o

On On

On r- NC

CT) OO

r- cm o o o d

On On

^ ^ ^ ^

no no" d d

O On 00 r-

m o >n ^ oo in -cf © 'ct •ct — ;

oi CM CM CM ■'t Tf

z z z z

sees

h m q h NO NO NO CM

<L>

>

oi &

<u

1)

c

• ^4

e

o

c

<u

s

(U

<u

<u

3

s

o

c

43

s

4)

<u

<L>

d d

> U

c* a

6 g

3 cO

Cu

u

64

Figure 1. Map of the study area. Stars indicate sites where R. cataractae has been vouchered (data taken from the Illinois Natural History Survey Fish Collec¬ tion, Champaign) and circles designate those sites where we failed to collect the dace during our survey.

SCALE

65

BOOK REVIEW 2012 -#1

Colorado's Spanish Peaks Region: An Exploration Guide to History, Natural History, Trails and Drives by Dr. Richard Keating. 350 pp. The Missouri Botanical Garden Press, St. Louis, MO. USD $24.99 ISBN 9781930723856.

If a person were to require one book about this area of southeastern Colorado as an introduction to its physical and historic marvels, that book should be the newly-released Colorado’s Spanish Peaks Region: An Exploration Guide to History, Natural History, Trails, and Drives by Richard Keating.

Perhaps it is our “orphan” status that keeps this area from so-called complete guides to geology, trails, history, etc., of Colorado, even those claiming to cover the Front Range or the Sangre de Cristos. If covered at all, Huerfano County usually merits a paragraph or two, possibly a page at most, and it is not always accurate information. In one fairly recent publication, for instance, the author could not tell the Devil’s Stairsteps from Pro¬ file Rock, with a picture of one and the name of the other beneath it.

Keating’s book, all 350 pages of it, is nearly all about Huerfano County, though it includes a bit of information about the region from the Great Sand Dunes National Park clear east to the dinosaur tracks of the Purgatoire Valley. Perhaps five percent of the text covers Las Animas County, which is just the way Huerfanos like it!

The author explains that his purpose for writing the book was the paucity of information about the Spanish Peaks area and his goal to fill that vacuum. He and his late wife have been regular visitors for some years, and so he has, alas, fallen in with the local disregard for correct names. Thus, the Cucharas river, pass and valley have become the Cuchara, and Mt. Mestas evolves into Mt. Maestas. These localisms in no way detract from the text.

Colorado's Spanish Peaks Region covers historic, geologic, and ecologic topics, giving a synopsis of inhabitants and travelers from prehistoric to frontier times, answering the common question of how the stone dykes were formed and other geologic mysteries (complete with a glossary of terms), offering guidelines on how to avoid predators, “real and rumored.” (No, he says, there are no wolves or grizzlies.) A very simple and straightforward explanation of why higher altitudes have higher radiation levels and thus different vegetation is offered. An understatement, at least this week, tells us “wind... is rather constant in this region,” giving fair warning to lady tourists worried about their coiffures and “local small pet advisories.”

Keating describes 12 scenic drives, from the all-paved Highway of Legends to the four- wheel-drive Medano Pass. Again, warnings are included to the uninitiated to the weather and its effect on dirt roads, noting road “conditions vary year by year” (a nice thing to say when often, they vary hour by hour). He further explains that much of the territory these roads cross is private property and goes so far as to include the state regulations on trespassing.

66

His thoughtful coverage of these tours includes a mile by mile itinerary of scenic, geo¬ logic, recreation, historic and biologic highlights, explaining how the monumentless Monument Lake got its name and even why not to go on a side road from Mosca Pass (because it only leads to a microwave radio tower).

A respectful explanation for our roadside memorials, called descansos in Spanish, is given and is an education in itself.

There are 20 hiking trails described here with their conditions and more warnings about weather - which cannot be overstressed in the mountains - trespassing, and rules of eti¬ quette such as keeping pets on leashes and packing out trash. He cautions readers about the occasionally difficult task of locating trailheads, and sometimes, even trails. His list of 10 basic necessities to take on a hike include a map and or compass to tackle some of these more elusive paths. One of these trails is the Baker Creek, which he says, “gives a close-up view of nature. Few area residents know it exists.” He also tells us there is construction trash dumped there. Keating includes the North Fork trail as a leisurely three-and-half hour walk with spectacular views and a less rigorous grade than some of the others, like the old road to the Bullseye mine on the West Spanish Peak.

He thoughtfully adds a section of topographic maps to show the roads and trails covered in the text.

Keating dedicates more than 100 pages to the descriptions and locations of flora and fauna, from prairie to peaks. Interesting trivia is included, such as the use by native tribes of a certain type of pine for lodgepoles, and thus the meaning of the tree’s name. At the end of the book, a separate index is provided for plant and animal names.

The author provides an extensive list of books, maps, pamphlets and other resources where more information about the Spanish Peaks area may be found. This is subtitled “The wood pulp section. Read so that trees won’t have died in vain.” This might give the impression that the section is extraneous, but it isn’t. Current web site addresses are included.

Keating’s text is accompanied by sharp photos, recent photos [!], of scenes and living things, to illustrate his points of why so-and-so is scenic, or a hard climb, or a unique plant. He shares his knowledge of photography so the reader can take equally clear pic¬ tures.

All in all, this is a fine book. The author deserves all congratulations for fulfilling his self-avowed aim of writing a comprehensive guide book to the seldom mentioned Span¬ ish Peaks area. Bravo!

Review by Nancy Christofferson. Reprinted with permission from Huerfano World Jour¬ nal 127(15): 7. Editor: Gretchen Lorr.

67

2012 ISAS STUDENT GRANT AWARD RECIPIENTS

Kelly Commons - Millikin University

How Does Wild Bird Feeding Affect the Wild Bird Community?

Madeline Knott - Millikin University

Comparison of IL-6 Levels and Staphylococcus aureus BKA in Central Illinois Songbirds

Grant Woods - Eastern Illinois University

Geospatial Investigation of the Occurrence of Confluent Meander Beds in East Central Illinois

Amanda Wildenberg - Eastern Illinois University

Fish, Macroinvertebrate, and Mussel Community Responses to Sanitary Effluent in an Illinois River

Kayla Little - Southern Illinois University - Edwardsville Avian Hematozoa of Illinois and Their Impacts on Health

Kate Koepp - Southern Illinois University - Edwardsville

Habitat Preferences of Fundulus notatus and F. olivaceus in an Illinois Stream Contact Zone

2012 BOTANY TRAVEL GRANT RECIPIENTS

Jennifer O’Brien - Eastern Illinois University

Roxane Krutsinger - Southern Illinois University - Edwardsville

68

69

2012 ANNUAL MEETING STUDENT PRESENTATION AWARDS

BOTANY DIVISION

Oral Presentations

1st Place: Eun Sun Kim - University of Illinois - Chicago

Investigating the Role of Genetic Diversity and Pollination Biology as Potential Causes for Reproductive Failure in Asclepias lanuginosa

Margaux Leja - University of Illinois - Urbana-Champaign Presence and Intensity of Pre-Dispersal Seed Predation in Response to Habitat Quality

2nd Place: Shanna David - Illinois College

Survival of Platanthera holochila (Orchidaceae) Seedlings In Vitro and Following Reintroduction in Hawaii

Poster Presentations

1st Place: Alex Sutton - Southern Illinois University - Edwardsville

Determining the DNA Content of a Unique Population of Schoenoplectus hallii in Howell County, MO

2nd Place: Roxane Krutsinger - Southern Illinois University - Edwardsville Rising Productivity Through Time in a Tallgrass Prairie Restoration

Jonathan Clark - Southern Illinois University - Edwardsville Germination of Field-Collected Japanese Hops ( Humulus japonicus, Cannabinaceae): Lab Techniques and Between Population Variation

CELL, MOLECULAR, & DEVELOPMENTAL BIOLOGY DIVISION

Oral Presentations - Undergraduate

1st Place: Bhargay Patel - Southern Illinois University - Edwardsville

Preservation of Beta-Cell Function by Peroxynitrite Scavenger Under Chronic Nutrient Overload

Oral Presentations - Graduate

1st Place: Stephanie B. Vernier - Southern Illinois University - Edwardsville Role of Lipids and Peroxynitrite in Beta Cell Defects Under Chronic Nutrient Overload

John M. Anderson - Eastern Illinois University

Protective Mechanisms Against Water Stress Evaluated in Insect Cells

70

■

71

CELL, MOLECULAR, & DEVELOPMENTAL BIOLOGY DIVISION - CONT’D

Poster Presentations - Undergraduate

1st Place: Ya-Lin Lu - Knox College

Diverse MicroRNAs in Dictyostelid Social Amoebas

Poster Presentations - Graduate

1st Place: Andrew J. Martens - Southern Illinois University - Edwardsville

UVA Induction of Lysosomal Membrane Permeabilization in Human Leukemia Cells, HL60

CHEMISTRY DIVISION

Oral Presentations

Outstanding: Brianna Norris - Western Illinois University

Eco- and User-Friendly Procedure for Oxidative Cleavage of Alkenes: Optimization and Development of an Undergraduate Experiment

Jennifer Tournear - Western Illinois University A Study of the Stability of Biopolymer-Reinforced Emulsion Systems Containing Lipophilic Bioactive Ingredients

Kamal Madavarapu - Governors State University Using Photoactivated Nanoparticles as Photodynamic Therapeutic Agent to Treat Prostate Cancer

Poster Presentations

Outstanding: Amanda M. Nappa & Benjamin M. Brown - Greenville College Synthesis of Aminoallenes

Abhilash Thamsetti, Sri-harika Talluri, and Shanque Ries -

Western Illinois University

Development of a Convenient and Pedagogically Useful Green Oxidation Protocol for Wider Adaptation in Undergraduate Laboratory Curriculum

COMPUTER SCIENCE DIVISION

Oral Presentations

1st Place: Jacob Thurman - Western Illinois University

Enhancing the Levenshtein String Edit Distance Algorithm to Identify Source Code Clones

72

73

EARTH SCIENCE DIVISION

Poster Presentations

1st Place: Kristina Pourtabib - Eastern Illinois University

Hermosa Formation, Silverton, CO, Sediment Source Area Petrographic Analysis

ENVIRONMENTAL SCIENCE DIVISION

Oral Presentations

1st Place: Yangus Zhu - Southern Illinois University - Edwardsville

Microbial Volatilization of Selenium by Pseudomonas sp. (STL-6) Isolated from the Soil - Stanleya pinnata System

Poster Presentations

1st Place: Kathryn Quesnell - Knox College

The Paleomicrobiology and Geomicrobiology of the Dakhleh Oasis, Egypt with Scanning Electron Microscopy

HEALTH SCIENCES & MICROBIOLOGY DIVISION

Oral Presentations

1st Place: Jennifer Galloway - Aurora University

Using Health Science Questionnaire to Survey the Influence of the Internet on Self Diagnosis by College Staff and Students: Cyberchondria Affects Healthcare Decisions

Poster Presentations - Health Sciences

1st Place: Whitney Malmborg - Aurora University

The Life Expectancy of Citizens in Kane County is Significantly Lower than Citizens of Illinois of the United States

Poster Presentations - Microbiology

1st Place: Kristopher Janezic - Eastern Illinois University

Phenotypic and Genotypic Characterization of Escherichia coli Isolated from Surface Waters in Illinois and Missouri

PHYSICS, MATHEMATICS, & ASTRONOMY DIVISION

Oral Presentations

1st Place: Jeff Carlson - Western Illinois University

Modeling Rormaldehyde Maser Variability with Easy Java Simulations

Poster Presentations

1st Place: Daniel Halbe - Western Illinois University

Arecibo Calibrators for Galactic Observations

74

75

ZOOLOGY DIVISION

Oral Presentations - Undergraduate

1st Place: Jason Scott - Southern Illinois University - Edwardsville

The Influence of Habitat Continuity on Genetic Structure Among Populations of Two Closely Related Topminnow Species

Oral Presentations - Graduate

1st Place: Kat Koepp - Southern Illinois University - Edwardsville

Movement Patterns in Two Closely-Related Species of Topminnow in a Southern Illinois Contact Zone

Poster Presentations - Undergraduate

1st Place: Mitchell Elting - Southern Illinois University - Edwardsville

Growth and Development of the Raccoon ( Procyon lotor ) Mandible

Poster Presentations - Graduate

1st Place: Adam Doelling - Southern Illinois University - Edwardsville

The Effect of Repeated Freezing on Oxidative Damage and Survival in the Freeze-Tolerant Goldenrod Gall Fly, Eurosta solidaginis

76

NOTES

NOTES

New York Botanical Garden Liorary

3 5

85 00342 6093

Transactions Information

Transactions Editor

Teresa (Tere) North Western Illinois University terenl956@gmail.com

Executive Secretary

Robyn Meyers Illinois State Museum rmyers@ museum .state .il .us

DIVISION CHAIRS

Agriculture

Vacant

Anthropology & Archaeology

Bonnie W. Styles Illinois State Museum styles@museum.state.il. us

Botany

Barbara Carlsward Eastern Illinois University bscarlsvvard@eiu.edu

Cell, Molecular, & Developmental Biology

Tom Fowler

Southern Illinois University Edwardsville tfowler@siue.edu

Chemistry

Dean Campbell Bradley University Campbell @ bumail .bradley .edu

Computer Science

Jim McQuillan Western Illinois University jm-mcquillan@wiu.edu

Earth Science

Jim Riley

Eastern Illinois University jdriley@eiu.edu

Engineering & Technology

Vacant

Environmental Science

Elizabeth Walton

Southern Illinois University Edwardsville elwalto@siue.edu

Health Sciences & Microbiology

Vance McCracken

Southern Illinois University Edwardsville vmccrac @ siue .edu

Physics, Mathematics, & Astronomy

Casey R. Watson Millikin University crwatson@millikin.edu

Sdence,Mathematics, & Technology Education

Kelly Barry

Southern Illinois University Edwardsville kbarry@siue.edu

Zoology David Duvernell

Southern Illinois University Edwardsville dduvern@siue.edu Paul Brunkow

Southern Illinois University Edwardsville pbrunko@siu.edu

Transactions of the Illinois State Academy of Science Volume 105 (2012)

CONTENTS

Academy of Science Editorial Information . . . inside front cover

Agriculture Papers

Mondyagu, Suneetha, David E. Kopsell, Richard W. Steffen, Dean A. Kopsell, and Robert L. Rhykerd

The Effect of Nitrogen Level and Form on the Growth and Development of Vetiver

Grass (Chrysopogon zizanioides)

1

Botany Papers

McClain, William E. and John E. Ebinger

Flora of Twin Shelters and Twin Mounds Hill Prairies, Pere Marquette State Park, Jersey County, Illinois, Changes Since 1963 . 11

Microbiology Papers

Cosgrove Pohren, Adonica S. and Scott M. Holt

Influence of Culture Conditions on Hydrogen Peroxide Production by Lactobacillus jensenii . 19

Zoology Papers

Bade, Simon R., F. Agustm Jimenez, Aaron K. Poole, and George A. Feldhamer

Prevalence of Baylisascaris procyonis and Implications for Reintroduced Woodrat

Populations in Southern Illinois . 33

Brohard, Justin, Marianne Robertson, and Anne Rammelsberg

Efficacy of Hoodia gordonii Extract as a Weight Loss Supplement: A Comparative Study Between an Invertebrate, Tenebrio molitor (Coleoptera: Tenebrionidae), and a

Vertebrate, Rattus norvegicus (Rodentia: Muridae) . 41

Huschen, Max S. and David Joseph Horn

Mass Death of Wintering American Robins (Turdus migratorius) in Decatur, Illinois 147

Tiemann, Jeremy S., Christopher A. Taylor, and Jason Knouft

Conservation Review of the Longnose Dace Rhinichthys cataractae (Valenciennes) in Northwestern Illinois . 57

Book Reviews

Christofferson, Nancy

Colorado’s Spanish Peaks Region: An Exploration Guide to History, Natural History, Trails and Drives, by Dr. Richard Keating . 65

Announcements

2012 Student Research Awards and Grants . 67

2012 Botany Travel Grant Recipients . 67

2012 Annual Meeting Student Awards . 69

Contributor Information . inside back cover

& .

...

XT

£?3?

J, (Of no- V4

law

Trans

ILLINOIS STATE ACADEMY OF SCIENCE

VOLUME 105 (2012) #3&4, pp. 77-156

Illinois State Academy of Science Founded 1907

Affiliated with the Illinois State Museum

ISSN 0019-2252 Printed by authority of the State of Illinois Springfield, Illinois

Transactions Information

Transactions Editor

Teresa (Tere) North Western Illinois University terenl956@gmail.com

Executive Secretary

Robyn Meyers Illinois State Museum rmyers@museum.state.il .us

DIVISION CHAIRS

Agriculture

Vacant

Engineering & Technology

Vacant

Anthropology & Archaeology

Bonnie W. Styles Illinois State Museum stvles@museum.state.il. us

Environmental Science

Elizabeth Walton

Southern Illinois University Edwardsville elwalto@siue.edu

Botany

Barbara Carlsward Eastern Illinois University bscaiisward@eiu.edu

Health Sciences

Vance McCracken

Southern Illinois University Edwardsville vmccrac@siue.edu

Cdl, Molecular, & Developmental Biology Tom Fowler

Southern Illinois University Edwardsville tfowler@siue.edu

Microbiology

Vance McCracken

Southern Illinois University Edwardsville vmccrac @ siue .edu

Chemistry

Dean Campbell Bradley University Campbell @ bumail .bradley .edu

Physics, Mathematics, & Astronomy

Casey R. Watson Millikin University crwatson@millikin.edu

Computer Science Jim McQuillan Western Illinois University jm-mcquillan@wiu.edu

Earth Science Jim Riley

Eastern Illinois University jdriley@eiu.edu

Science, Mathematics, & Technology Education

Kelly Barry

Southern Illinois University Edwardsville kbarry@siue .edu

Zoology David Duvernell

Southern Illinois University Edwardsville dduvern @ siue .edu

Paul Brunkow

Southern Illinois University Edwardsville pbrunko@siu.edu

77

DEBUT OF MANUSCRIPT SUBMISSION AND PUBLICATION ANNOUNCED

Transactions Goes Digital

In conjunction with the 2013 Annual Meeting Transactions will be moving to an online publishing format at a new website http://www.ilacadofsci.com.

This provides many benefits, including the 4 Es.

• Ecology: Less paper wasted as papers are submitted online and reviewed electronically

• Economy: Reduced postage and printing costs to prevent increased publication charges and makes color photos/figures possible

• Efficiency: Papers published as approved, rather that semi-annually, resulting in a decreased time from submission to publication

• Expediency: Credit card payments available with no surcharge made possible via secure PayPal

Check the INFORMATION FOR CONTRIBUTORS on the back cover that provides some new information for authors

In addition, the repository of prior manuscripts is also being moved to the new website http://www.ilacadofsci.com starting with the most current issues. Manuscripts will be indexed by title, author, and year making searching easy by even partial titles and individual authors. The projected timeline for making all manuscripts available at the new site and avail able for PDF download is as follows

• April 2013 - 2000 through 2012 and 2013 articles as published

• June 2013 - 1980 through 1999

• August 2013 - 1908 through 1939

• October 2013 - 1940 through 1959

• December 2013 - 1960 through 1979

http://www.ilacadofsci.com

Transactions of the Illinois State Academy of Science (2012) Volume 105, #3&4 pp. 79-84

received 2/21/12 accepted 11/7/12

The Varying Phenology and Growth Patterns of Baptisia bracteata (Fabaceae)

in Reconstructed Prairie

Chris E. Petersen, Sharon M. Bauzys, Felicia A. Speranske, and Barbara A. Petersen

College of DuPage, Glen Ellyn, IL 60137

ABSTRACT

First flowering time and flowering duration of Baptisia bracteata Muhl. ex. Ell (cream wild indigo), were examined in the Russell R. Kirt Tallgrass Prairie, Glen Ellyn, IL, dur¬ ing 2008, 2009, and 201 1. While some B. bracteata delay flowering, most bloom during May, just prior to the blooming period of the taller and more prolific congener, B. alba. Both congeners are host to the pre-dispersal seed predator, Apion rostrum Say (Apio- nidae), although the reproductive cycle of the weevil is synchronized to B. alba. Factors relating to seed yield and the growth of the rhizomatous B. bracheata were also investi¬ gated. B. bracteata with more shoots, flowers and racemes tended to flower earlier and longer than those with fewer counts, evidence of the importance of plant size to flowering phenology. Still, as indicated by flower count/plant, a larger size was only positively correlated to seeds matured/plant during 2011. Counts of A. rostrum showed positive relationships to pods and seeds matured/plant during 2008 and 2011. Mean shoot count/plant was not significantly different between 2008 and 2011 among 39 tracked individuals. Growth was largely limited to smaller individuals while larger individuals sometimes declined in size. Selective pressures favoring larger size may include competi¬ tion for resources. However, rewards of seeds matured/plant based on a larger inflo¬ rescence size, may not be realized every year. Larger plants may be more prone to resource limitations and weevil attacks. Variances in plant sizes can be just explained by age. Alternatively, size plasticity may occur due to changing environmental conditions and reproductive effort from year to year. It remains unknown if low output of mature pods was due to a lack of pollinators. By subsequently having lower seed output, the spe¬ cies may lower its' attractiveness to A. rostrum.

INTRODUCTION

One basic principle of life history theory explains that there is a cost to reproduction which is minimized by trade-offs among components of reproductive yield (Sanchez- Humanes, et al. 2011; Williams, 1966). Flowering phenology may be under the influ¬ ences of trade-offs within a species, e.g., when the timing of first flowering or flowering duration show asynchrony as a response to changing environmental constraints (Elzinga et al., 2007). Larger plants are often found to bloom earlier and longer than smaller plants, presumably to attract pollinators and because they have greater nutrient reserves available for flowering (Bolmgren and Cowan, 2008; Bustamante and Burquez, 2008; Pettersson, 1994). By flowering off-peak, smaller plants may be able to avoid consumers

80

such as seed predators, but perhaps at the cost of less favorable weather (Albrectsen, 2000; Evans et al., 1989; Mahoro, 2002; McIntosh, 2002; Mduma et al., 2007).

Baptisia bracteata Muhl. ex. Ell (cream wild indigo; = B. leucophaea) is a native tail- grass perennial of the Midwest which shows variance in flowering phenology. At our study site, the reconstructed Russell R. Kirt Prairie, IL, the species blooms in May when a new season of emergent prairie growth has just begun to show (Petersen et al., 1998). Most plants tend to flower at the same time while a few delay. Using data collected dur¬ ing 2008, 2009, and 2011, we investigated factors that could explain the asynchrony. We also examined factors relating to seed yield and the change in size of 39 plants over a 4- year span.

Each B. bracteata consists of subterranean rhizomes from which multiple aerial shoots develop to form a concentric cluster measuring up to a half meter in diameter. Racemes along shoots radiate outward from the cluster, exposing conspicuous displays of yellow flowers. Aging flowers along the indeterminate racemes shift from being staminate to carpellate. This shift allows for a degree of self pollination (Haddock and Chaplin, 1982). Bombus are the major pollinators. Pods inflate with pollination and can bear over 14 seeds. Unlike the congener, B. alba L. (Vent)(white wild indigo) which also grows in the prairie, inflated pods of B. bracteata remain attached regardless of seed number, and the ones that fail to mature seeds appear to ripen at the same rate as those that retain seeds. B. alba often aborts many to all of its inflated pods in apparent response to environmental extremes and low counts of seeds in pods (Petersen et al., 201 1).

At our study site, B. bracteata is infested by the pre-dispersal seed predator, Apion ros¬ trum Say (Apionidae). The weevil also infests and is more synchronized to pod develop¬ ment of the later blooming B. alba (Petersen and Wang, 2006). Overwintering adult wee¬ vils oviposit into inflating pods of both congeners. Resulting larvae consume seeds as their only source of nutrition. Maturation is reached by August, and the weevils disperse when pods dehisce or fragment.

METHODS

Reconstruction of the 7.1 ha Russell R. Kirt Tallgrass Prairie began in 1984. Today, the dominant tall grasses are Andropogon gerardii Vitman (big bluestem), Sporobolus heterolepsis Gray (prairie dropseed), and Sorghastrum nutans (L.) Nash (Indian grass). Over 100 species of forbs contribute to the flora. The prairie was burned annually until 2006. It has not been burned since.

Sampling methods in 2008, 2009, and 2011 followed the same protocol. A concentric cluster of B. bracteata , with shoots being within 10 cm of one another, was assumed to be one individual based on soil excavation of individuals not otherwise included in sam¬ pling. Fifty to 51 plants were randomly selected for sampling each year. First and last flowering dates of each B. bracteata were recorded along with counts of shoots, racemes, and flowers. When pods matured, they were counted along with their contents of seeds and weevils. If pods had holes from which weevils could escape, A rostrum count/plant was prorated using mean counts from intact pods. Shoot and flower counts per plant pro-

81

vided alternative measures of plant size. Seeds matured/plant and counts of A. ros- truml plant provided measures of reproductive output and weevil infestation, respectively.

All statistical analyses were done using Statistica 6 (StatSoft, 2001). Due to failure in meeting assumptions of parametric testing, relationships between flowering phenology, other reproductive allocations to include seeds matured, and weevil infestation were explored using Spearman Rank Correlation (Zar, 1984). Rhizomatous growth was esti¬ mated by comparing the ratio of 2011:2008 shoot counts to 2008 shoot counts using Spearman Rank Correlation.

RESULTS

The first B. bracteata flowered on May 11th during 2008 and 2011, and on May 26th during 2009. Plant measurements according to year are presented in Table 1. For individuals, an earlier flowering start, a longer flowering duration, and a higher flowering count/plant were positively correlated for all years, except in 2011 between flowering initiation and duration (Table 2). However, as indicated by flower count/plant, a larger size was only positively correlated to seeds matured/plant during 2011. Plants producing more flowers also tended to show higher shoot counts; but more shoots did not correlate consistently with an earlier time and duration of flowering. It also did not translate to a higher yield of seeds matured. Finally, mean counts of A. rostrum/plani were positively correlated to pods and seeds matured per plant in 2008 and 2011, but not in 2009.

Table 1. Summary (mean ± standard error; n) of variables associated with reproductive yield of Baptisia bracteata and infestation by Apion rostrum according to year. Flower¬ ing time is in reference to the mean date plants flowered after the first B. bracteata bloomed.

Variable		Year
2008	2009	2011
Time of first flowering	9.6 ±0.8 (51)	9.2 ±0.8 (50)	14.0 ± 1.0 (42)
Flowering duration (days)	16.8 ±0.6 (51)	17.2 ±0.5 (50)	16.5 ± 1.1 (42)
Shoots/plant	8.0 ± 1.0 (51)	9.6 ±0.9 (50)	10.9 ± 1.3 (42)
Flowers/plant	114.4 ± 16.2 (51)	95.1 ±11.9 (50)	122.9 ±23.8 (42)
Pods matured/plant	9.8 ±3.1 (51)	32.3 ± 6.4 (50)	19.2 ±5.9 (42)
Seeds matured/plant	28.1 ± 16.3 (51)	62.5 ±20.6 (50)	8.5 ±6.5 (42)
Apion rostrum! plant	10.8 ±4.4 (20)	13.6 ±4.3 (32)	26.7 ±8.3 (24)
% of Baptisia bracteata bearing mature pods	37.3% (51)	80.0% (50)	59.5% (42)

While shoot count among tracked B. bracteata did not change significantly from 2008 to 2011 (t = 1.876; df = 38; P < 0.068), plants with fewer shoots accounted for most increases (Figure 1). The ratio in shoot count between 2011 and 2008 was negatively correlated with shoot count in 2008 (rs = -0.54; P < 0.05). From 2008 to 201 1 , 22 plants showed an increase in shoot count, 4 showed no change, while 13 decreased in shoot count.

82

Table 2. Spearman rank correlation comparing variables of flowering phenology, reproductive yield, and Apion rostrum infestations presented sequentially for years 2008, 2009, and 2011. AC = Counts of Apion rostrum/plant. The AC sample sizes (n's) for 2008, 2009, and 2011, were 20, 32, and 24, respectively. Otherwise, n = 51 for 2008, 50 for 2009, and 42 for 2011. Bold type denotes significance (P < 0.05).

	FD	SC	FC	PM	SM	AC
Time of first flowering (FF)	-0.85	-0.21	-0.36	<-0.00	0.23	-0.33
	-0.55	-0.35	-0.55	-0.28	0.02	-0.17
	-0.23	-0.07	-0.32	<0.00	0.17	-0.08
Flowering duration (FD)		0.13	-0.85	-0.06	-0.20	0.30
		0.36	0.43	0.52	0.20	-0.12
		0.45	0.54	0.39	0.43	0.14
Shoot count/plant (SC)			0.65	0.15	-0.06	0.21
			0.41	0.43	0.10	0.12
			0.42	0.44	0.27	0.51
Flower count/plant (FC)			0.21	<0.00	0.43
			0.59	0.20	0.31
			0.48	0.42	0.42
Pods matured/plant (PM)				0.56	0.52
				0.52	0.34
				0.72	0.78
Seeds matured/plant (SM)					0.56
					0.25
					0.43

DISCUSSION

Flower count per B. bracteata provided a better correlating factor to flowering phenology than shoot count/plant. The discrepancy may reflect the additional function of shoots in photosynthesis. Shoot number may vary due to disproportionate allocations of energy to non-reproductive and reproductive structures as a reaction to changing environmental conditions. Regardless, results are consistent with larger B. bracteata blooming earlier and longer than those smaller.

Plant size can indicate age. Alternatively, plant size may be influenced by the local environment and changes in reproductive effort from year to another. With larger B. bracteata being more likely to become smaller, and smaller individuals larger between 2008 and 2011, such changes in size can function to promote the fitness of a perennial over the course of its lifetime (Salguero-Gomex and Casper, 2010). Larger size may offer competitive benefits to acquisition of resources not investigated in this study. Neverthe¬ less, greater rewards of seeds matured/plant, where plant size is based on flower count, may not be realized every year. Larger plants may also be more prone to resource limita¬ tions and attack by consumers like A. rostrum (Hainsworth et al., 1984; Ohasi and Yahara, 2000). A longer term study should offer insights in explaining size differences in B. bracteata.

83

Figure 1. Shoot count ratio of B. bracteata comparing years 2011 to 2008, plotted against the 2008 shoot count. Best-fit linear regression line is provided to help visualize trend.

Despite the seemingly prolific flowering by B. bracteata , few pods were matured/plant. It remains unknown if pollen limitations are due to a lack of pollinators at the reconstructed site. Mature pods bear seeds. By having a seed output much lower than B. alba (Petersen et al., 2007), the species may be less attractive to A. rostrum explaining the synchroniza¬ tion of the weevil’s reproductive cycle around the taller and more prolific congener.

LITERATURE CITED

Albrectsen, B. R. 2000. Flowering phenology and seed predation by a tephritid fly: Escape of seeds in time and space. Ecoscience 7: 433-438.

Bolmgren, K. and P. D. Cowan. 2008. Time-size tradeoffs: a phylogenetic comparative study of flowering time, plant height and seed mass in a north-temperate flora. Oikos 1 17:424-429. Bustamante, E. and A. Burquez. 2008. Effects of plant size and water on the flowering phenology of the Organ Pipe Cactus ( Stenocereus thurberi). Ann. Botany 102:1019-1030.

Elzinga, J. A., A. Atlan, A. Biere, Luc Gigord, A. E. Weis, and G. Bernasconi. 2007. Time after time: flowering phenology and biotic interactions. Trends Ecol. Evol. 22:432-439.

Evans, E., C. C. Smith, and R. P. Gendron. 1989. Timing of reproduction in a prairie legume: sea¬ sonal impacts of insects consuming flowers and seeds. Oecologia 78:220-230.

Haddock, R. C., and S.J. Chaplin. 1982. Pollination and seed production in two phenologically divergent prairie legumes ( Baptisia leucophaea and B. leucantha). Am. Midi. Nat. 108: 175-186.

84

Hainsworth, F. R., L.L. Wolf, and T. Mercier. 1984. Pollination and pre-dispersal seed predation: net effects on reproduction and inflorescence characteristics in Ipomopsis aggregate. Oecologia 63:405-409.

Mduma, S. A., A. R. E. Sinclair, and R. Turkington. 2007. The role of rainfall and predators in determining synchrony in reproduction of savanna trees in Serengeti National Park, Tanzania. J. Ecol. 95:184-196.

Mahoro, S. 2002. Individual flowering schedule, fruit set, and flower and seed predation in Vaccin- ium hirtum Thunb. (Ericaceae). Can. J. Bot. 80:92-92.

McIntosh, M. E. 2002. Flowering phenology and reproductive output in two sister species of Ferocactus (Cactaceae). Plant Ecol. 159:1-13.

Ohashi, K. and T. Yahara. 2000. Effects of flower production and predispersal seed predation on reproduction in Cirsium purpuratum. Can. J. Bot. 78:230-236.

Petersen, C.E., S. C. Lindsey, D. M. Dudgeon, and R. A. Pertell. 1998. The effect of seed predation on pod abortion by the prairie legume, Baptisia leucophaea. The Trans. Ill. State Acad. Sci. 91:47-52.

Petersen, C. E. and W. Wang. 2007. Congener host selection by the pre-dispersal seed predator, Apion rostrum (Coleoptera: Apionidae). Gt. Lakes Entomol. 39:68-74.

Petersen, C. E., A. A. Mohus, B. A Petersen, and B. A. McQuaid. 2010. Multiyear study of factors related to flowering phenology and reproductive yield of Baptisia alba in Northeastern Illinois. Trans. Ill. State Acad. Sci. 103:109-1 17.

Pettersson, M. W. 1994. Large plant size counteracts early seed predation during the extended flowering season of a Silene uniflora (Caryophyllaceae) population. Ecography 17:264-271 .

Salguero-Gomex R. and B. B. Casper. 2010. Keeping plant shrinkage in the demographic loop. J. Ecol. 98: 312-323.

Sanchez-Humanes, B. V. L. Sork, J. M. Espelta. 2011. Trade-offs between vegetative growth and acorn production in Quercus lobata during a mast year: the relevance of crop size and hierar¬ chical level within the canopy. Oecologia 166:101-1 10.

StatSoft. 2001. Statistica AX 6.0. 2001. StatSoft, Tulsa, OK.

Williams, G. C. 1966. Natural selection costs of reproduction and a refinement of Lack's Principle. Am. Nat. 100:687-690.

Zar, J.H. 1984. Biostatistical Analysis, 2nd ed. Prentice-Hall, Inc., Englewood Cliffs, NJ, USA.

Transactions of the Illinois State Academy of Science (2012) Volume 105,#3&4 pp. 85-93

received 5/2/12 accepted 1/7/13

Vascular Flora of Capel Glacial Drift Hill

Prairie Natural Area,

Shelby County, Illinois

William E. McClain1, John E. Ebinger2*, Roger Jansen3, and Gordon C. Tucker2

'Illinois State Museum, Springfield, IL 62706 department of Biological Sciences, Eastern Illinois University, Charleston, IL 61920

^Illinois Department of Natural Resources 1660 West Polk Avenue, Charleston, IL 61920 Corresponding author (jeebinger@eiu.edu)

ABSTRACT

The vascular flora of Capel Glacial Drift Hill Prairie Natural Area, Shelby County, Illi¬ nois was studied during the 2009 and 2010 growing seasons. The 1.10 ha glacial drift hill prairie is located on a southwest-facing slope associated with Lake Shelbyville, Wolf Creek State Park 4 km east of Findley, Illinois. Plant community structure was deter¬ mined using m2 square quadrats located at one-meter intervals along two randomly located transect lines. Frequency, mean cover, relative frequency, relative cover, and importance value (I. V. total = 200) were determined from the data collected. A total of 106 vascular plant taxa were observed on the site, with 39 encountered in the plots. Andropogon gerardii (big bluestem) had the highest importance value followed by Schi- zachyrium scoparium (little bluestem), Echinacea pallida (pale coneflower), and Dalea purpurea (purple prairie clover). Exotic species were represented by six taxa.

Key Words: Andropogon gerardii, glacial hill prairie formation, soil slumping.

INTRODUCTION

Small prairie openings in the forested landscapes of east-central Illinois were first described and named “hill prairies” by Vestal (1918). These small prairies (mostly < 1 ha), now refer to as glacial drift hill prairies, developed near the crest of slopes or spurs on mostly Wisconsin age glacial till. In east-central Illinois these prairie openings have been studied in Coles County (Ebinger 1981, Reeves et al. 1978, Owens and Cole 2003, Behnke and Ebinger 1989), Vermilion County (Owens and Ebinger 2008) and on Illi- noian glacial till in Jasper County (Edgin et al. 2010). Glacial drift hill prairies have also been studied in central Illinois in Moultrie County (Owens et al. 2006), Macoupin County (McClain et al. 2002), and are not uncommon in the Illinois River valley from Peoria and Tazewell counties north through Marshall, Woodford, and Putnam counties (McFall and Karnes 1995).

Glacial drift hill prairies have many edaphic, physical, and floristic similarities. They occur on steep south- to southwest-facing slopes with well-drained soils that are low in organic content and nutrients (Ebinger 1981, McClain et al. 2002, Owens et al. 2006,

86

Owens and Ebinger 2008). The predominant vegetation is native warm season clump grasses complemented by prairie forbs with a sparse vegetation ground cover. Drying winds, unstable soil, fire, cutting, and grazing were thought to play significant roles in the development and maintenance of these sites (Vestal 1918, Reeves et al. 1978). The pre¬ sent study was undertaken to describe the composition and structure of the vegetation of Capel Glacial Drift Hill Prairie Natural Area. This prairie was added to the Illinois Natu¬ ral Areas Inventory in 1999 and is considered to be of high natural quality.

DESCRIPTION OF THE STUDY SITE

Capel Glacial Drift Hill Prairie Natural Area is located on a southwest-facing slope overlooking Lake Shelby ville in Wolf Creek State Park, 4 km east of Findley, Shelby County, Illinois (SW 1/4, NE1/4, Section 7, T12N, R5E). This 1.10 ha prairie is located at an elevation of about 190 m on the Shelby ville terminal moraine of Wisconsin glaciation in the Grand Prairie Section of the Grand Prairie Natural Division (Schwegman 1973). Tallgrass “black soil” prairie dominated the extensive flat to gently rolling uplands of this natural division, the forests being mostly restricted to floodplains and dissected topogra¬ phy of river valleys and other hilly areas (Anderson 1991, Ebinger and McClain 1991). Lake Shelbyville was formed when the Kaskaskia River was dammed just east of the town of Shelbyville in 1970 by the Army Corps of Engineers. Small prairie opening are occasionally encountered in the rough topography surrounding the lake where edaphic and microclimatic conditions combined to produce excessively droughty sites (Reeves et al. 1978, Edgin et al. 2010). The forest communities found at Wolf Creek State Park were discussed by Newell et al. (1991).

The soil of the glacial drift hill prairie is Miami loam, a highly eroded soil with little of the original A horizon present, the original loess deposits having eroded away (Gotsch 1996). This soil, which is on 18 to 30 % slopes, is well drained, low in organic content, slightly acid, and commonly low in available phosphorus and potassium. Many gravel size pebbles are imbedded in the soil. In this part of Illinois annual precipitation averages 97.5 cm, with April having the highest rainfall (9.4 cm). Mean annual temperature is 1 1 .8 C with July being the hottest month (average of 24.6 °C), the coldest being January (-3 °C). The average number of frost-free days is 171 (Midwestern Regional Climate Center 2011).

First examined in 1998 by personnel from the Illinois Department of Natural resources (Smithl998), Capel Glacial Drift Hill Prairie was first burned in the spring of 1999. Subsequent management efforts have focused on control of Melilotus albus (white sweet clover) and Securigera varia (crown vetch). Prescribed burning was conducted in April of 2005 and 2010, and a wild fire burned the site in the spring of 201 1 .

MATERIALS AND METHODS

Collections of the vascular flora of the prairie and adjacent open woodland margin were made periodically during the 2009 and 2010 growing seasons. Voucher specimens were collected for most taxa and deposited in the Stover-Ebinger Herbarium of Eastern Illinois University, Charleston (EIU). Nomenclature follows Mohlenbrock (2002), and the

87

assignment of exotic species status was determined using Taft et al. (1997) and Mohlen- brock (2002).

Quantitative sampling was conducted in late summer of 2010 using nr quadrats located at one-meter intervals along two randomly placed 25 m transects oriented perpendicular to the slope (n = 25 per transect). Even-number quadrats were placed to the right, odd- numbered to the left of each transect. Percent cover of each species was determined using Duabenmire cover class system (Daubenmire 1959) as modified by Bailey and Poulton (1968): class 1 = 0-1%; class 2 = 1-5%; class 3 = 5-25%; class 4 = 25-50%; class 5 = 50- 75%; class 6 = 75-95%; class 7 = 95-100%. From these data, frequency (%), mean cover (%), relative frequency, relative cover, and Importance Value [relative frequency + rela¬ tive cover) were determined.

The Floristic Quality Index (FQI) was determined using the coefficient of conservatism (CC) assigned to each species by Taft et al. (1997). As used here, the FQI is a weighted index of species richness (N) and is the arithmetic product of the mean coefficient of conservatism (C-value), multiplied by the square root of species richness (V N) of the site [FQI = C-value (VN)].

RESULTS

Floristic Composition

At total of 106 vascular plant species were collected or observed in and at the edge of the prairie (Appendix I). Of the species present 26 were monocots in five families while 80 were dicots in 31 families. Seven non-native (exotic) taxa were found with only Melilotus albus occurring in the plots. Predominant plant families were Asteraceae (24 species), Poaceae (19), and Fabaceae (10). No state endangered or threatened species were observed (Illinois Endangered Species Protection Board 2005). The C-value and FQI for all species were 4.54 and 46.7 respectively with 21 species having a CC of seven or more.

Ground Layer

Of the 106 species encountered on the prairie 39 were found in the plots (Table 1). Prairie grasses were the most important species. Andropogon gerardii (big bluestem) dominated with the highest importance value (I.V. of 38.5) and mean cover (24.57%), followed by Schizachyrium scoparium (little bluestem) (I.V. of 25.3 and a mean cover of 13.99%). Other species with importance values ±10.0 included Echinacea pallida (pale cone- flower), Dalea purpurea (purple prairie clover), Aster oolentangensis (sky-blue aster), and Asclepias viridiflora (green milkweed). Five woody species were recorded in the plots, Rosa Carolina (Carolina rose) with an I.V. of 6.1, Cornus drummondii (rough¬ leaved dogwood) with an I.V. of 1.3, and seedlings of three tree species ( Carya ovata, Quercus stellata, and Sassafras albidum).

88

Table 1. Frequency (%), mean cover (% of total area), relative frequency, relative cover, and importance value (I.V.) for the ground layer species at Capel Glacial Drift Hill Prai¬ rie Natural Area, Wolf Creek State Park, Shelby County, Illinois. (*exotics)

Species	Freq. %	Mean Cover	Rel. Freq.	Rel. Cover	I.V.
Andropogon gerardii	90	24.57	8.6	29.9	38.5
Schizachyrium scoparium	88	13.99	8.4	16.9	25.3
Echinacea pallida	98	11.92	9.3	14.4	23.7
Dalea purpurea	82	4.91	7.8	5.9	13.7
Aster oolentangensis	70	5.35	6.7	6.5	13.2
Asclepias viridiflora	84	2.78	8.0	3.4	11.4
Euphorbia corollata	68	1.81	6.5	2.2	8.7
Brickellia eupatorioides	56	1.56	5.3	1.9	7.2
Aster patens	38	2.76	3.6	3.3	6.9
Rosa Carolina	34	2.36	3.2	2.9	6.1
Lithospermum canescens	48	0.59	4.6	0.7	5.3
Lespedeza virginica	36	1.40	3.4	1.7	5.1
Coreopsis palmata	30	1.22	2.9	1.5	4.4
*Melilotus albus	34	0.56	3.2	0.7	3.9
Hypericum sphaerocarpum	34	0.47	3.2	0.6	3.8
Aureolaria grandiflora	14	1.33	1.3	1.6	2.9
Sporobolus cryptandrus	26	0.28	2.5	0.3	2.8
Solidago nemoralis	16	1.00	1.5	1.2	2.7
Silphium integrifolium	10	1.02	1.0	1.2	2.2
Carex pensylvanica	12	0.69	1.1	0.8	1.9
Cornus drummondii	6	0.61	0.6	0.7	1.3
Ruellia humilis	12	0.06	1.1	0.1	1.2
Comandra umbellata	10	0.10	1.0	0.1	1.1
Ipomoea pandurata	6	0.42	0.6	0.5	1.1
Physostegia virginiana	8	0.14	0.8	0.2	1.0
Pycnanthemum pilosum	6	0.37	0.6	0.4	1.0
Tree seedlings (3 species)	8	0.09	0.8	0.1	0.9
Elymus virginicus	4	0.02	0.4	—	0.4
Taenidia integerrima	4	0.02	0.4	—	0.4
Eupatorium serotinum	2	0.06	0.2	0.1	0.3
Helianthus divaricatus	2	0.06	0.2	0.1	0.3
Helianthus mollis	2	0.06	0.2	0.1	0.3
Antennaria plantaginifolia	2	0.01	0.2	—	0.2
Aster turbinellus	2	0.01	0.2	—	0.2
Galium circaezans	2	0.01	0.2	—	0.2
Poly gala senega	2	0.01	0.2	—	0.2
Scutellaria leonardii	2	0.01	0.2	—	0.2
Totals		82.63	100.0	100.0	200.0
Bare ground and litter		26.24

89

DISCUSSION

The glacial drift hill prairies of east-central Illinois are characterized by similar soils (type, structure, stability, slope, exposure), flora (common prairie grasses and forbs as well as few conservative prairie species), and bare ground (25 to 50% of the total cover), but commonly differ in size, species richness, and distribution and abundance of the dominant grass species. These differences are likely related to edaphic characteristics of the sites. Some of the glacial drift hill prairies, particularly those in Macoupin County (McClain et al 2002) and Jasper County (Edgin et al. 2010) are on more shallow slopes, are larger, and have higher species richness. In these hill prairies Andropogon gerardii commonly dominates and the soils are stable, soil slumping being uncommon. The gla¬ cial drift hill prairies of the Middle Fork of the Vermilion and the upper Embarras Rivers (Coles and Vermilion Counties), in contrast, are usually on steeper slopes, tend to be somewhat smaller, and have lower species richness. Also, Schizachyrium scoparium and Sorghastrum nutans are the most abundant grasses while Andropogon gerardii is appar¬ ently a minor component (Edgin et al. 2010). In these hill prairies soil slumping is com¬ mon.

Slumping, the down-hill movement of large masses of soil, is a characteristic of many glacial drift hill prairies in central Illinois. The sparse prairie vegetation can not hold the tremendous weight of the saturated clay soil on the steep slopes. In wet years, soil breaks away and slide down hill, taking prairie vegetation, shrubs, and sometimes large trees with it. On Capel Hill Prairie excessive soil slumping, due to shoreline erosion of Lake Shelbyville, has limited the size of the prairie. The slumping has helped maintain the hill prairie by eliminating surrounding trees and by creating bare ground which rapidly suc¬ ceeds to a community containing mostly prairie grasses and forbs (Owens et al. 2006). Slumping facilitates an early succession stage that seems to be favorable to Schizachyr¬ ium scoparium (Vestal 1918). This sun-loving, early to mid-successional species is usu¬ ally associated with drier sites with poorer soils and tends to decrease with succession.

Capel Hill Prairie is located 1 1 km southwest of Coneflower Glacial Drift Hill Prairie in Moultrie County (Owens et al. 2006). This prairie, which is located on the Cerro Gordo recessional moraine of Wisconsin glaciation, occurs on a moderate slope with eroded, well-drained Miami silt loam soil that is slightly acidic and has low organic content. On both Coneflower and Capel hill prairies Andropogon gerardii was the dominant grass while Schizachyrium scoparium ranked second among the grasses. Though similar, Capel Hill Prairie has low species diversity (106 taxa) and has excessive soil slumping due to its position on the Lake Shelbyville shoreline, Coneflower Hill Prairie, in contrast, has high species diversity (164 taxa), and minimal soil slumping. Excessive disturbances are probably responsible for some of this lack of diversity. It is very possible that previous to the formation of Lake Shelbyville, Capel Hill Prairie had minimal slumping. The undercutting of the shoreline by wave action is undoubtedly responsive for the present excess slumping, and may be responsible for the low species diversity. Five species origi¬ nally reported as occurring on this hill prairie (Smith 1998) ( Calystegia spithamaea, Helenium autumnale, Liatris aspera , Polygala verticillata, Pycnanthemum tenuifolium ) were not observed during the two years of our study.

90

ACKNOWLEDGEMENTS

The authors would like to thank the Illinois Department of Natural Resources for permis¬ sion to conduct the study, and particularly to District Heritage Biologist Eric Smith for

his help, advice, and encouragement. Dr. Loy R. Phillippe, Illinois Natural History Sur¬ vey, Champaign, Illinois, verified the identification of many of the species.

LITERATTURE CITED

Anderson, R. C. 1991. Presettlement forests of Illinois. Pp. 9-19. in: G. V. Burger, J. E. Ebinger, and G. S. Wilhelm (editors.). Proceedings of the oak woods management workshop. Eastern Illi¬ nois University, Charleston, Illinois.

Bailey, A. W. and C. E. Poulton. 1968. Plant communities and environmental relationships in a portion of Tillamook burn, northwestern Oregon. Ecology 49: 1-13.

Behnke, G. and J. E. Ebinger. 1989. Woody invasion of glacial drift hill prairies in east-central Illinois. Transactions of the Illinois State Academy of Science 82: 1-4.

Daubenmire, R. 1959. A canopy coverage method of vegetation analysis. Northwest Science 33: 43-64.

Ebinger, J. E. 1981. Vegetation of glacial drift hill prairies in east-central Illinois. Castanea 46: 115-121.

Ebinger, J. E. and W. E. McClain. 1991. Forest succession in the prairie peninsula of Illinois. Illi¬ nois Natural History Survey Bulletin 34:375-381.

Edgin, B., S. J. Adams, E. J. Stork, and J. E. Ebinger. 2010. Flora and species dominance of Green Prairie, Jasper County, Illinois - A glacial drift hill prairie on Illinoian till. Erigenia 23:24-33.

Illinois Endangered Species Protection Board. 2005. Checklist of Endangered and Threatened Ani¬ mals and Plants of Illinois. Endangered Species protection Board, Springfield, Illinois.

Gotsch, K. A. 1996. Soil Survey of Shelby County. U.S.D.A. Natural Resources Conservation Ser¬ vice and Illinois Agricultural Experiment Station. U.S. Government Printing Office, Washington DC.

McClain, W. E., M. A. Phipps, H. H. Eilers, and J. E. Ebinger. 2002. Vascular plants of glacial drift prairies in Macoupin County, Illinois. Castanea 67: 54-60.

McFall, D. and J. Karnes. 1995. (editors). A directory of Illinois Nature Preserves. Volume 2. Northwestern, Central and Southern Illinois. Illinois Department of Natural Resources, Spring- field, Illinois.

Midwestern Regional Climate Center. 2011. http://mcc.sws.uiuc.edu

Mohlenbrock, R. H. 2002. Vascular flora of Illinois. Southern Illinois University Press, Carbon- dale, Illinois.

Newell, S. A., L. L. Horton, and J. E. Ebinger. 1991. Vegetation of Wolf Creek State Park, Shelby County, Illinois. Erigenia 11:22-26. 1991.

Owens, N. L. and G. N. Cole. 2003. 25 years of vegetational changes in a glacial drift hill prairie community in east-central Illinois. Transactions of the Illinois State Academy of Science 96: 265-269. '

Owens. N. L. and J. E. Ebinger. 2008. Windfall Glacial Drift Hill Prairie, Vermilion County, Illi¬ nois: Present vegetation and changes since 1977. Transactions of the Illinois State Academy of Science 101: 157-165.

Owens, N. L., G. C. Tucker, and J. E. Ebinger. 2006. Flora and vegetation of Coneflower Glacial Drift Hill Prairie Natural Area, Moultrie County, Illinois. Rhodora 108:370-386.

Reeves, J. T., U. D. Zimmerman, and J. E. Ebinger. 1978. Microclimatic and soil differences between hill prairies and adjacent forests in east-central Illinois. Transactions of the Illinois State Academy of Science 71: 156-164.

Schwegman, J. E. 1973. Comprehensive plan for the Illinois Nature Preserves Commission: part 2. The Natural Divisions of Illinois. Illinois Nature Preserves Commission, Rockford, Illinois.

Smith, E. L. 1998. Nomination Form, Wolf Creek State Park - Capel Hill Prairie.

Taft, J. B., G. S. Wilhelm, D. M. Ladd, and L. A. Masters. 1997. Floristic quality assessment for vegetation in Illinois, a method for assessing vegetation integrity. Erigenia 15: 1-95.

91

Vestal, A. G. 1918. Local inclusions of prairie within forest. Transactions of the Illinois State Academy of Science 11: 12-126.

Appendix I. Vascular plant species encountered Capel Glacial Drift Hill Prairie Natural Area, Wolf Creek State Park, Shelby County, Illinois are listed alphabetical by family under the major plant groups. Collecting numbers (JEE) are listed after each species. Specimens are deposited in the Ebinger/Stover Herbarium (EIU), Eastern Illinois Univer¬ sity, Charleston. Species followed by INAI (Illinois Natural Areas Inventory) were observed on the site during the original survey by the INAI. During the present study most of these species were observed. (*exotics)

DICOTS

Acanthaceae

Ruellia humilis Nutt.:33032 Apiaceae

Taenidia integerrima (L.) Drude:33074 Asclepiadaceae

Asclepias purpurascens L.:32912 Asclepias tuberosa L.:32407 Asclepias viridiflora Raf.:3291 1

Asteraceae

Achillea millefolium L.:32913 Ambrosia artemisiifolia L.:32689 Antennaria plantaginifolia (L.) Hook.:32760 Arnoglossum atriplicifolium (L.) H.

Robins. :32409

Aster oolentangiensis Riddell:32690 Aster patens Ait.:32384 Aster turbinellus Lindl.:32385 Brickellia eupatorioides (L.) Shinners:32410 Coreopsis palmata Nutt.:3241 1 Echinacea pallida (Nutt.) Nutt.:32412 Erigeron strigosus Muhl.:32914 Eupatorium altissimum L.:32691 Eupatorium serotinum Michx.:32692 Helenium autumnale L.:INAI Helianthus divaricatus L.:32413 Helianthus mollis Lam.TNAI (observed) Liatris aspera Michx. :INAI Parthenium integrifolium L.:32414 Ratibida pinnata (Vent.) Barnh.:33075 Silphium integrifolium Michx.:32415 Solidago nemoralis Ait.:32386 Solidago speciosa Nutt.:32693 Solidago ulmifolia Muhl.:32437 Verbesina helianthoides Michx.:32416 Boraginaceae

Lithospermum canescens (Michx.) Lehm.:32756

Caprifoliaceae

Viburnum prunifolium L.:32694 Viburnum rafinesquianum Schultes:33031

Cistaceae

Lechea tenuifolia Michx.: 324 17 Convolvulaceae

Calystegia spithamaea (L.) Pursh:INAI Ipomoea pandurata (L.) G.F.W. Mey.:32418

Cornaceae

Cor nus drummondii C.A. Mey.:32439 Cornus florida L.:32766

Ebenaceae

Diospyros virginiana L.:32440 Elaeagnaceae

*Elaeagnus umbellata Thunb.:32696 Euphorbiaceae

Euphorbia corollata L.: 324 19 Fabaceae

Dalea Candida (Michx.) Willd.:32441 Dalea purpurea Vent.: 32420 Desmodium sessilifolium (Torr.) Torr. & Gray.: 32442

Lespedeza capitata Michx. :32698 *Lespedeza cuneata (Dum.-Cours.) G. Don:32699

Lespedeza violacea (L.) Pers.:32700 Lespedeza virginica (L.) Britt.: 32421 *Melilotus albus Medic. :32422 *Melilotus officinalis (L.) Pallas:32915 *Securigera varia (L.) Lassen: 32697

Fagaceae

Quercus muhlenbergii Engelm.:32701 Quercus stellata Wangh.:32916

92

Gentianaceae

Frasera caroliniense Walt.:32755 Hypericaceae

Hypericum sphaerocarpum Nutt.:32423 Juglandaceae

Carya tomentosa (Poir.) Nutt.:33076 Lamiaceae

Hedeoma pulgioides (L.) Pers.: 32702 Monarda bradburiana Beck.:32703 Physostegia virginiana (L.) Benth.:32704 Pycnanthemum pilosum Nutt.:32424 Pycnanthemum tenuifolium Schrad.:INAI Scutellaria leonardii Epling:32443

Lauraceae

Sassafras albidum (Nutt.) Nees:INAI (observed)

Oxalidaceae

Oxalis violacea L.:32759 Polyalaceae

Polygala senega L.:32444 Polygala verticillata L.:INAI

Portulacaceae

Claytonia virginica L.:32761 Primulaceae

Lysimachia lanceolata Walt.:32917 Ranunculaceae

Ranunculus fascicularis Bigel:32758 Rhamnaceae

Ceanothus americanus L.:32425 Rosaceae

Amelanchier arborea (Michx. F.) Fern.:32763 Rosa Carolina F.:32445

Rubiaceae

Galium circaezans Michx. :32426 Houstonia lanceolata (Poir.) Britt. :32446

Santalaceae

Comandra umbellata (F.) Nutt.:INAI (observed)

Scrophulariaceae

Agalinus tenuifolia (Vahl.) Raf.:32388 Aureolaria grandiflora (Benth.) Pennell :32427 Penstemon pallidus Small:32428

Violaceae

Viola pedata F.:32764 Viola pratincola Greene:32757

Vitaceae

Parthenocissus quinquefolia (F.) Planch. :INAI (observed)

MONOCOTS

Cyperaceae

Carex brachyglossa Mack.:33030 Carex hirsutella Mack. :32905a Carex muhlenbergii Schk.:32429 Carex pensylvanica Fam.:32762

Iridaceae

Sisyrinchium albidum Raf.: 32765 Filiaceae

Allium canadense F.:32905 Orchidaceae

Spiranthes magnicamporum Sheviak:32389 Poaceae

Agrostis gigantea Roth:32380 Agrostis hyemalis (Walt.) BSP.:32906 Bromus pubescens Muhl.:32907 Andropogon gerardii Vitman:32430 Danthonia spicata (F.) Roem. & Schultes:32705

Dichanthelium acuminatum (Sw.) Gould & Clark:32706

Dichanthelium depauperatum Muhl.:32908 Elymus x ebingeri G.C. Tucker:32382 Elymus canadensis F.:32432 Elymus histrix F.: 3 2707 Elymus virginicus F.:32381 Festuca subverticillata (Pers.) E.B.

Alexeev:32909 *Poa compressa F.:32433 *Poa pratensis F.:INAI (observed) Schizachyrium scoparium (Michx.) Nash:32434

Sphenopholis obtusata (Michx.) Scribn.:33910 Sporobolus cryptandrus (Torr.) Gray:32448 Sporobolus heterolepis (Gray) Gray:32708 Tridens flavus (F.) Hitchc.:32709

Transactions of the Illinois State Academy of Science (2012) Volume 105,#3&4 pp. 93-99

received 5/19/12 accepted 10/24/12

The Effects of Beaver-created Wetlands on Surface Water Quality of Lotic Habitats in Northern Illinois

Eric K. Bollinger and Brian Conklin

Department of Biological Science, Eastern Illinois University, Charleston, Illinois 61920

ABSTRACT

We examined 41 chemical parameters in water samples taken upstream and downstream from 7 beaver-created wetlands in DuPage County, IL. Overall, a significant majority of parameters (18 of 26) present in detectable amounts had lower concentrations in down¬ stream samples, suggesting that these wetlands do serve to improve water quality by filtering out some pollutants. However, only 5 of these parameters had statistically significant differences between upstream and downstream samples (manganese, arsenic, sulfate, total solids, and calcium) and 2 of these actually had higher concentrations down¬ stream (manganese and arsenic). This suggests that materials used by beavers in dam construction (e.g., cornstalks, treated lumber) may also serve to reduce certain aspects of water quality.

INTRODUCTION

Wetlands may act as natural filters, removing dissolved and solid pollutants from aquatic ecosystems (e..g., Whigham et al. 1988, Agovina 1990, Verhoeven et al. 2006). For example, artificial wetlands (especially those associated with wastewater treatment) remove significant quantities of various compounds including nitrogen, phosphorus, total suspended solids, and sulfates (Gersberg et al. 1984, 1986, Bowmer 1987, Breen 1990, Shutes 2001, Jiang et al. 2007, Pankratz et al. 2007). Data from natural wetlands also indicate positive effects on downstream water quality, although these effects are often complex (Richardson 1985, Whigham et al. 1988, Montreuil and Merot 2006, Verhoeven et al. 2006).

Beaver-created wetlands are one type of natural wetland that could also improve water quality in downstream areas. For example, Parker et al. (1985) found reduced concentra¬ tions of nitrogen, suspended solids, and phosphorus downstream from beaver wetlands. Other studies have also indicated that beaver ponds may reduce downstream transport of sediments, nutrients, and pollutants (e.g., Naiman and Melillo 1984, Francis et al. 1985, Maret et al. 1987, Cirmo and Driscoll 1993).

In this study, 41 chemical parameters were examined from water taken above and below beaver-created wetlands in DuPage County, Illinois. This study was designed to examine the effects that these urban populations of beavers had on water quality of the associated streams and rivers.

94

MATERIALS AND METHODS

Study Area

This study was conducted on lands owned by the Forest Preserve District of DuPage County in northeastern Illinois. This preserve system encompasses more than 10,100 hec¬ tares in 60 preserves. Numerous marshes, rivers, and streams within this preserve system support beaver populations.

Site Selection

Beaver wetlands used in this study were selected based on the following criteria. The site had to (1) have an active beaver colony that was maintaining a dam, (2) be located on land owned or managed by the Forest Preserve District of DuPage County, (3) be associ¬ ated with a lotic ecosystem with clearly delineated single points of inflow and outflow, and (4) have no close association with other wetlands. Seven beaver-created wetlands were found that met these criteria.

Water Sample Collection

We collected surface water samples in 500 ml high-density polyethylene, wide-mouthed bottles that had been washed with 1 N HCL to remove possible contamination. One sam¬ ple was taken upstream and one downstream from each beaver wetland. “Upstream” sam¬ ples were collected approximately 25 m upstream from the impact area of the wetland where significant surface water movement was still occurring. “Downstream” samples were collected at least 5 m but no more than 20 m downstream from the beaver dam. We took all samples from the middle-third of the stream or river.

Analysis of Chemical Parameters (Techniques used for the first two sampling dates,

27 June and 10 July 1991)

The concentration of dissolved calcium (Ca++) was determined using the EPA-approved EDTA titrimetric method with Eriochrome Black T as the indicator (APHA 1985). Concentrations of dissolved chloride (Cf) were determined using reagents obtained from Hach Company (Lovelend, CO) utilizing the EPA-approved silver nitrate buret titration method using 0.0141 N AgN03 (Hach Company 1989). Dissolved nitrate (N03‘) concentrations were determined using a hybrid method that utilized Hach’s NitraVer 5 nitrate reagent powder pillow. This reagent contains the necessary chemicals to apply the cadmium reduction method for photometric determination of nitrate. The NitraVer 5 was added to 25 ml of the water sample in test tubes that had a path length of 2.5 cm after which samples were analyzed spectrophotometrically (Spec 21) to determine optical den¬ sity at a wavelength of 400 nm. Samples of known nitrate concentration were used to construct a concentration vs. transmittance curve. Applying this curve, the concentrations of the samples could be determined. Dissolved sulfate (S04~2) concentrations were deter¬ mined with a hybrid method using Hach’s SulfaVer 4 powder pillow and test tubes with a 2.5cm path length. Optical density at 450 nm was determined using a spectrophotometer (Spec 21) and a standard curve was constructed utilizing a stock sulfate solution (APHA 1985).

Methods used for the Entire Study. Total phosphorus (tP) concentrations were determined using Standard Methods’ Ascorbic Acid Method and persulfate digestion. The pH was determined using a d02 meter and a Yellow Springs Instrument Company probe. Total

95

solids (TS) were determined by using a 50 ml porcelain drying dish into which 40 ml of the water sample were added. We dried the sample in an oven overnight at 98°C, then for an additional 15 min at 1 10°C and then cooled the sample in a desiccator for one hr. The samples were then weighed to the nearest 0.0001 g. Total non-volatile solids were deter¬ mined by ashing the TS sample in a 510°C muffle furnace for 15 min and then weighing it after it had cooled. Total volatile solids (TVS) were determined by taking the mathematical difference between TS and TNVS.

Methods used for the last 6 sampling dates. Concentrations of dissolved arsenic (As) and dissolved cadmium (Cd) were determined using inductively coupled plasma/mass spectrometry (ICP/MS; method 6010-EPA-5w846). Other dissolved metal concentrations were determined by inductively coupled plasma-optical emission spectroscopy (ICP- OES; method 6020-EPA-5w846). Beginning with the sample collected 16 July 1991 (the third sample), dissolved sulfate, dissolved chloride, dissolved nitrate, dissolved fluoride, and dissolved bromide concentrations were determined using ion chromatography (IC; method 300-Mcaww-EPA).

The EPA-approved ICP/MS, ICP-OES, and IC methods were performed by Dave Roberts of the Enesco Rocky Mountains Analytical Laboratory in Arvada, CO. Samples were sent by overnight express mail in a 0°C cooler and preserved with nitric acid upon receipt by the lab. Table 1 summarized the parameters we examined, the method utilized for analy¬ sis, and the detection limits for each.

Table 1. Water quality parameters examined from water samples collected at beaver-cre¬ ated wetlands in DuPage County, IL. Detection limits in ppm except pH.

Parameter (symbol)	Detection limit	Detection Method
Aluminum (Al)	0.009	ICP-OES
Silver (Ag)	0.002	ICP-OES
Arsenic (As)	0.001	ICP/MS
Barium (Ba)	0.003	ICP-OES
Beryllium (Be)	0.002	ICP-OES
Boron (B)	0.005	ICP-OES
Bromide (Br-)	0.2	IC
Cadmium (Cd)	0.001	ICP/MS
Calcium (Ca)	0.1/0.01	Titrimetric/ICP-OES
Chloride (C1-)	5/3	AgN03 Titrimetric/OC
Chromium (Cr)	0.006	ICP-OES
Cobalt (Co)	0.004	ICP-OES
Copper (Cu)	0.004	ICP-OES
Iron (Fe)	0.007	ICP-OES
Fluoride (F-)	0.1	IC
Lead (Pb)	0.01	ICP-OES
Lithium (Li)	0.002	ICP-OES
Magnesium (Mg)	0.008	ICP-OES
Manganese (Mn)	0.003	ICP-OES
Molybdenum (Mo)	0.005	ICP-OES

96

Parameter (symbol)	Detection limit	Detection Method
Nickel (Ni)	0.004	ICP-OES
Nitrate (N03 )	0.1	NitraVer 5/IC
Dissolved Phosphorus (dP)	0.09	ICP-OES
Total Phosphorum (tP)	0.1	Ascorbic Acid
Potassium (K)	0.2	ICP-OES
Antimony (Sb)	0.02	ICP-OES
Selenium (Se)	0.05	ICP-OES
Silica Oxide (Si02)	0.2	ICP-OES
Strontium (Sr)	0.02	ICP-OES
Tin (Sn)	0.03	ICP-OES
Sodium (Na)	0.1	ICP-OES
Sulfate (S042 )	5	IC
Thallium (Tl)	0.5	ICP-OES
Titanium (Ti)	0.003	ICP-OES
Vanadium (V)	0.004	ICP-OES
Zinc (Zn)	0.002	ICP-OES
Total Solids (TS)	25	Standard Methods
Total Volatile Solids (TVS)	25	Standard Methods
Total Non-volatile Solids (TNVS)	25	TS-TVS
PH	0.1 units	Field pH meter
Dissolved Oxygen (d02)	0.1	YSI meter and probe

Data Analysis

We sampled all 7 wetlands for the first two sampling dates (27 June and 10 July 1991), after which the dam associated with one wetland was destroyed. Water levels became low enough to preclude sampling in 3 other wetlands after the second sampling date as a result of a drought. Because of this situation and the fact that certain methods varied between the first two and last 6 sampling dates, we analyzed our data as 2 separate bal¬ anced data sets. The first data set (“data set 1”) consisted of 7 wetlands sampled over two dates. The second data set (“data set 2”) consisted of 3 wetlands sampled over the last 6 sampling dates (for most parameters) and all 8 sampling dates for calcium, chloride, nitrate, sulfate, pH, dissolved oxygen, and total solids. We used repeated-measures ANOVA to analyze both data sets, with sampling date as the repeated measure. Chemical parameters were analyzed separately. However, the following chemical parameters were typically below detection limits and so were excluded from the statistical analyses: silver (Ag), beryllium (Be), chromium (Cr), cobalt (Co), lithium (Li), lead (Pb), antimony (Sb), selenium (Se), tin (Sn), thallium (Tl), titanium (Ti), and vanadium (V).

RESULTS

Data set 1 (7 wetlands, 2 dates)

We failed to find any significant differences between upstream and downstream concentrations for this data set (ANOVA, all F < 3.75, P > 0.05), although sulfate concentrations were marginally lower downstream (F = 3.74, P = 0.10).

97

Data set 2 (3 wetlands, 6 dates)

Five parameters showed significant differences between upstream and downstream concentrations (ANOVA, P < 0.05, Table 2). Three parameters (S04 2, TS, and Ca) had lower average concentrations downstream and two (Mn and As) had higher average concentrations downstream. Overall, 18 parameters had lower average concentrations downstream, 8 had higher concentrations downstream and one (pH) was the same (chi- square = 3.85, df = 1 , P < 0.05).

Table 2. Comparison of upstream and downstream concentrations of water quality parameters for 3 beaver-created wetlands over 6 sampling dates. All concentrations are in ppm (except pH); see Table 1 for key to parameter abbreviations.
Parameter	Mean Upstream Concentration	Mean Downstream Concentration	Percent Differ¬ ence	Sig. of F-Statistic
Mn	0.0205	0.3281	+1500*	0.02**
As	0.022	0.050	+ 127	0.02**
so42-	114.7	85.5	-25	0.04**
TS	1590	1254	-21	0.05**
Ca	114.4	98.6	-14	0.09**
Fe	0.0292	0.0920	+215	0.16
A1	0.0267	0.0519	+94	0.22
Cl-	258	317	+23	0.024
Na	259.8	197.4	-24	0.27
Mg	55.08	47.88	-13	0.32
Mo	0.0125	0.0105	-16	0.40
Zn	0.0121	0.0092	-24	0.40
tP	0.5194	0.6694	+29	0.41
F-	1 .3429	1.1095	-17	0.41
Sr	0.5205	0.4248	-18	0.53
B	0.3240	0.2902	-10	0.58
Ba	0.1920	0.1629	-16	0.59
no3	1 .0343	1.2014	+ 16	0.65
K	6.7571	6.2095	-8	0.67
Cu	0.009	0.008	-11	0.67
Br-	0.2571	0.2381	-7	0.73
Ni	0.0041	0.0039	-5	0.73
do2	5.2	5.1	-2	0.73
dP	0.6743	0.6114	-2	0.76
PH	7.4	7.4	0	0.83
Cd	0.018	0.016	-11	0.84
Si02	4.9429	4.9571	+0	0.99

*‘+’ denotes higher concentration downstream; denotes lower concentration downstream **P< 0.10, ANOVA

98

DISCUSSION

Wetlands are thought to act as biological filters, removing dissolved and solid pollutants and improving water quality (Feeney and Morrell 1985, Hair 1986, Agovina 1990, Verhoeven et al. 2006). Thus, our prediction was that beaver wetlands would improve water quality by lowering concentrations of pollutants in our downstream samples. How¬ ever, most parameters did not differ significantly upstream and downstream from beaver wetlands. Of those parameters that did differ 2 of the 5 (Mn and As) actually had higher concentrations downstream. Arsenic used to be a common preservative added to treated lumber, so if beavers imported treated lumber in the construction of their dams, leaching from this material may explain our results. Given the proximity of our beaver wetlands to human development, this is certainly possible. However, we did not notice any type of finished lumber in the beaver dams in this study, and neither chromium nor copper were similarly elevated downstream (other elements used in treated lumber; Lebow et al. 1999), so the explanation of this result remains problematic. However, arsenic was used in pesticides prior to 1993, and can be taken up by corn in measurable concentrations (Parsons et al. 2008). In addition, manganese is found in corn, and beavers are known to commonly eat corn in the Midwest. Furthermore, we have seen cornstalks used in beaver dam construction.

Although our small sample of wetlands probably contributed to our lack of significant results, we did find that a significant majority of parameters had lower concentrations downstream. This suggests that our beaver-created wetlands do serve to filter out some pollutants. However, items brought into the aquatic system by beavers (e.g., corn, treated lumber?) may also serve to reduce water quality downstream from these wetlands.

ACKNOWLEDGEMENTS

We would like to thank the Forest Preserve District of DuPage County, Illinois and East¬ ern Illinois University for funding, Benedictine University for lab space and equipment, Michael Redmer for field assistance, Dave Roberts of Enseco Rocky Mountain Analyti¬ cal Lab for chemical analyses, and D. R. Ludwig, C. P. Pederson, and D. Zimmerman for manuscript review.

LITERATURE CITED

Agovina, A. V. 1990. Wetland identification: a means to prevent potential public health problems. Journal of Environmental Health 52:280-281.

APHA AWWA WPCF. 1985. Standard Methods for the Examination of Water and Wastewater, 16lh ed., Washington, D.C.

Bowmer, K. H. 1987. Nutrient removal from effluents by an artificial wetland: influence of rhizo- sphere aeration and preferential flow studied using bromide and dye tracers. Water Research 21:591-599.

Breen, P. F. 1990. A mass balance method for assessing the potential of artificial wetlands for wastewater treatment. Water Research 24:689-697.

Cirmo, C. P. and C. T. Driscoll. 1993. Beaver pond biogeochemistry: acid neutralizing capacity generation in a headwater wetland. Wetlands 13:277-292.

Feeney, P. and B. Morrell. 1985. Wetlands disposal is sewage option. American City & County March 1985:91-92.

99

Francis, M. M., R. J. Naiman, and J. M. Melillo. 1985. Nitrogen fixation in beaver ( Castor canadensis ) influenced subarctic streams. Hydrobiologia 121:193-202.

Gersberg, R. M., B. V. Elkins, and C. R. Goldman. 1984. Use of artificial wetlands to remove nitro¬ gen from wastewater. Journal of Water Pollution Control Federation 56:152-156.

Gersberg, R. M., B. V. Elkins, S. R. Lyon, and C. R. Goldman. 1986. Role of aquatic plants in wastewater treatment by artificial wetlands. Water Research 20:363-368.

Hach Company. 1989. Water analysis handbook. Hach Company, Loveland, CO.

Hair, J. D. 1986. Who cares about wetlands? EPA Journal 12:14-15.

Jiang, C., X. Fan, G. Cui, and Y. Zhang. 2007. Removal of agricultural non-point source pollutants by ditch wetlands: implications for lake eutrophication control. Hydrobiologia 581:319-327.

Lebow, S. T., D. O. Foster, and P. K. Lebow. 1999. Release of copper, chromium, and arsenic from treated southern pine exposed in seawater and freshwater. Forest Produces Journal 49:80-89.

Maret, T. J., M. Parker, and T. E. Fannin. 1987. The effect of beaver ponds on the nonpoint source water quality of a stream in southwestern Wyoming. Water Research 21:263-268.

Montreuil, O. and P. Merot. 2006. Nitrogen removal in valley bottom wetlands: assessment in headwater catchments distributed throughout a large basin. Journal of Environmental Quality 35:2113-2122.

Naiman, R. J. and J. M. Melillo. 1984. Nitrogen budget of subarctic stream altered by beaver ( Cas¬ tor canadensis ). Oecologia 62:150-155.

Pankratz, S., T. Young, H. Cuevas-Arellano, R. Kumar, R. F. Ambrose, and I. H. Suffet. 2007. The ecological value of constructed wetlands for treating urban runoff. Water Science and Technol¬ ogy 55:63-69.

Parker, M., F. J. Wood, Jr., B. H. Smith, and R. G. Elder. 1985. Erosional downcutting in lower order riparian ecosystems: have historical changes been caused by the removal of beaver? In Riparian Ecosystems and their Management: Reconciling Conflicting Uses. (Edited by R. R. Johnson, C. D. Ziebell, D. R. Patton, P. F. Ffolliott, and R. H. Hamre), pp. 35-38. First North American Riparian Conference; April 16-18; Tucson, Ariz. Gen. Tech. Rep. RM-120. USDA, Forest Serv., Rocky Mt. Forest and Range Expt. Station. Ft. Collins, CO.

Parsons, J. G., A. Martinez-Martinez, J. R. Peralta- Videa, and J. L. Gardea-Torresday. 2008. Speciation and uptake of arsenic accumulated by corn seedlings using XAS and DRC-ICP-MS. Chemosphere 70:2076-2083.

Richardson, C. J. 1985. Mechanisms controlling phosphorus retention capacity in freshwater wet¬ lands. Science 226:1424-1427.

Shutes, R. B. E. 2001. Artificial wetlands and water quality improvement. Environment Interna¬ tional 26:441-447.

Verhoeven, J. T. A., B. Arheimer, C. Yin, and M. H. Hefting. 2006. Regional and global concerns over wetlands and water quality. Trends in Ecology and Evolution 21:96-103.

Whigham, D. F., C. Chitterling, and B. Palmer. 1988. Impacts of freshwater wetlands on water quality: a landscape perspective. Environmental Management 12:663-671.

100

Transactions of the Illinois State Academy of Science (2012) Volume 105,#3&4,pp. 101-105

received 5/17/12 accepted 1 1/23/12

Discovery of New Localities for the Threatened Kirtland’s Snake ( Clonophis kirtlandii ) in Central Illinois

Angelo P. Capparella1', Todd Springer2, and Lauren E. Brown1 'School of Biological Sciences, Illinois State University Campus Box 4120, Normal, IL 61790-4120 2 1 835 E. Lafayette St., Bloomington, IL 61701 "Correspondence: apcappar@ ilstu .edu

ABSTRACT

Three localities (two new, one reconfirmed) in central Illinois with habitat and behavioral observations are described for Kirtland’s snake which is quite rare throughout its range in the U.S.A.

INTRODUCTION

The Illinois State Threatened Kirtland’s snake Clonophis kirtlandii (Kennicott, 1856) is one of the rarest snakes in the Midwest because of the nearly complete destruction of native prairie, its primary former habitat. This small snake is semifossorial and tends to occupy crayfish burrows (Anton et al., 2003) in damp, open areas. The species can be easily identified by its red or orange belly bordered by a single row of round black spots on each side (Phillips et al., 1999). The purpose of the red belly is unknown. It is questionable if aposematic coloration is involved as the snake is not known to be poison¬ ous and is reluctant to bite. When first captured, the snake flattens its body like a thin ribbon. Known natural food includes earthworms, slugs, water striders, leeches, and cray¬ fish (Conant, 1943; Minton, 1944; Bavetz, 1993; Thurow, 1993).

The range of C. kirtlandii was probably best shown in Conant and Collins (1998, map p. 305), although new records have extended its range southward into northwestern Tennes¬ see (Frymire and Scott, 2012). The species has been found from eastern Missouri to west¬ ern Pennyslvania, and from northeastern Illinois and southern Michigan to northwestern Tennessee. In Illinois its range encompassed much of the central and northeastern por¬ tions of the State. Most of the distribution in Illinois and elsewhere occurred within the historic Prairie Peninsula (Transeau, 1935). By the 1890s the species had become rare in the northern half of Illinois (Garman, 1892). Brown et al. (1975) suggested that the populations in Illinois may be relictual and that the species might be becoming extinct in the western portion of its range. Since 1975, a few additional relictual localities have been reported in Illinois (e.g., Brown, 1987), always based on only one or a few speci¬ mens. The purpose of this paper is to report two new records and the confirmation of a nearly half-century old record in central Illinois with habitat notes and behavioral observations.

102

NEW AND RECONFIRMED LOCALITY RECORDS

Living specimens were captured, photographed, and released unharmed at the following three localities in southern McLean County.

1 . NW 14, NE !4, Sec. 26, T22N, R2E, Randolph Township, 4 km (2.5 mi) NE Hey worth. One individual was captured and photographed by T. Springer 16 October 2010 in a small grassy area adjacent to a small creek fed by a spring. The creek is a tributary of Kickapoo Creek. The snake was in leaf litter and nearby there were crayfish burrow chimneys. Old field-savannah is north of the site and a nearly pristine mature woodland occurs on a hillside south of the creek. When an attempt was made to pick up the snake, it flattened its body. Another sighting was made in the summer of ca. 2001 in mowed grass about 20 m (65.6 ft) north of the creek.

2. SE 14, NE 14, Sec. 19, T22N, R1E, Funks Grove Township, 2.4 km (1 .5 mi) SW Funks Grove. One individual was caught and photographed 6 June 2004 by E. Smith and A. Funk in an old field (former pasture) adjacent to the parking lot of the Sugar Grove Nature Center. A small intermittent creek and outhouse are located near the capture site. Subsequent inspection of this creek revealed crayfish burrow chimneys. Another C. kirtlandii was seen by R. Carriger on 12 September 201 1 along a small intermittent creek that receives flow from the aforementioned creek. Another sighting by S. Marshall occurred on 20 September 2011, not far from the September 12 sighting along this second intermittent creek. This snake was photographed by J. Tobias and measured at 38 cm (14.9 in) in total length. It was moving in a shaded area near a group of tree stumps and logs approximately 9 m (29.5 ft) from the creek and not far from a playground. The last documented sighting was 20 May 2012. It was found in a mulch pile by a young girl near the Marshall site and photographed by M. Litwiller. Habitats in the nearby area include restored prairie, old field in various successional stages, streams, former pasture or tilled areas, and open woodland.

3. SE '4, SW !4, Sec. 35, T23N, R4E, Dawson Township, 7 km (4.4 mi) NE LeRoy. K. Harness caught one individual and photographed it on 9 April 2011. Another capture was photographed on 14 August 2011. Both were found under rocks a considerable distance from each other (ca. 222 m [728 ft]) at the dam for Dawson Lake in Moraine View State Recreation Area. Both were estimated to be 20.3-25.4 cm (8-10 in) in total length. The specific habitat at the two sightings is under riprap for erosion control at the dam. A road runs across the dam with dense forest and cleared areas nearby. Numerous other potential habitats (marshes, streams, pond, old fields, former pasture, other prairie-like habitats) occur in the area.

DISCUSSION

Numerous species of snakes in eastern North America flatten their body when alarmed (Conant and Collins, 1998). This behavior is particularly well developed in C. kirtlandii in both frequency and thinness of body. Often this is interpreted as a defensive reaction to make the body appear larger in dorsal view, and hence more threatening to a potential predator. The body flattening has also been noted by Tucker (1994) to enhance the ability of C. kirtlandii to utilize small subterranean crevices. At the Randolph Township site.

103

T.S. attempted to pick up the newly encountered C. kirtlandii which was exhibiting flattening behavior. However, this proved to be rather difficult because T.S. was wearing gloves at the time. Thus, another potential function of flattening behavior may be to inhibit a potential predator from picking up and devouring the C. kirtlandii.

There are two other historical localities known for McLean County: (1) Normal: Garman (1892), no voucher specimen; UIMNH 4962, no other information; BMNH 93.1.2.1-2 (Conant 1943); and (2) Lake Bloomington: Holman (1966), no voucher specimen. In addition, there is an older record for Dawson Lake in Moraine View State Recreation Area, ISUC 703, 5 May 1966, J.A. Holman collector. It is likely that the Normal popula¬ tion is extinct because of intense urbanization. However, the Lake Bloomington region encompasses a relatively large area that has numerous habitats which might be suitable for and inhabited by C. kirtlandii. We do not know the exact location of Holman’s record from Dawson Lake.

Bavetz (1993, 1994) indicated there was a total of 70 known locality records for C. kirtlandii in Illinois. He surveyed 19 of these sites, and 14 other sites near known locali¬ ties that appeared to have appropriate habitat to support the species. Living C. kirtlandii were seen at only two of the old sites and only one new locality was found. Bavetz (1994) suggested that only eight localities “support reproducing populations” based on voucher specimens or photographic slides. In light of Bavetz’s findings, our report of living C. kirtlandii sighted multiple times at two new localities and one older locality in McLean County take on considerable significance because of the Threatened status of the species in Illinois.

In the species description, Kennicott (1856: 96) commented briefly on early habitat of C. kirtlandii in northern Illinois: “The few specimens obtained have been found in the woods, generally under logs.” Garman (1892: 275) commented further on early habitat in Illinois: “A handsome snake, which ten years ago was not uncommon along prairie brooks, in the central part of the State. Tiling, ditching, and cultivation of the soil have destroyed its haunts and nearly exterminated it... I have never seen it elsewhere than on the open prairie.”

Mid-20th century habitat for C. kirtlandii was described in Conant’s (1943: 328) mono¬ graph as: “essentially an inhabitant of open country... does occur in woods, but it is far more abundant in prairie-like situations.” Later, Conant’s (1958) field guide indicated that the species usually occurs in wet meadows.

We suggest that many present habitats of C. kirtlandii may be primarily successional and degraded derivatives (including extremely disturbed urbanized areas) of historical wet prairies. In non-urban regions they may be most frequently found in small, open grassy areas that are wet or adjacent to small watercourses or ponds. Some might also be found in wooded areas (possibly old fields). Presence of crayfish burrows or previously con¬ structed burrows of other animals are also of importance (Tucker, 1994).

The primary cause of the decline of C. kirtlandii is probably the pervasive destructive effects of agriculture on the snake’s habitat, with urbanization being a secondary cause. However, there are many possible factors that have had a role in the global decline of

104

reptiles (Gibbons et al., 2000). Two or more of these may have acted together in a detri¬ mental manner causing the decline of C. kirtlandii.

Clonophis kirtlandii is classified as a State Threatened Species in Illinois and Ohio, and State Endangered in Indiana, Kentucky, Michigan and Pennsylvania (Ernst and Ernst, 2003; Gibson and Kingsbury, 2004). The U.S. Fish and Wildlife Service (Dodd et al., 1985) placed C. kirtlandii in its Category 2 (possibly appropriate for federal listing as Endangered or Threatened but more information may be needed). Furthermore, there is apparently no conservation program specifically targeting C. kirtlandii anywhere throughout its range (Gibson and Kingsbury, 2004).

McLean County might be an appropriate county to initiate a conservation program, as four of the records (our three plus Lake Bloomington area ) occur in areas with suitable, or potentially suitable habitat (nature preserves, undeveloped land owned by the City of Bloomington, sympathetic land owner). Furthermore, the occurrence of two or more sightings at each of the three localities we report suggests the existence of extant popula¬ tions. Another locality occurs in Woodford County near the McLean County line in a pasture but otherwise relatively undisturbed “wet meadow” (Brown, 1987). Other coun¬ ties that have a number of locality records (Champaign, Cook) may also have several areas suitable for conservation programs of C. kirtlandii. If conservation programs are not soon activated by governmental agencies or the public, this small innocuous snake may become extinct.

ACKNOWLEDGMENTS

We thank: C.A. Phillips (Illinois Natural History Survey) for the loan of UIMNH 4962; J.R. Brown, E.L. Mockford, and two anonymous reviewers for critically reading the ms.; E. Smith and A. Funk for providing information on the Funks Grove locality and photo¬ graphs of the specimen; R. Carriger and S. Marshall for their separate reports of recent sightings; J. Tobias and M. Litwiller for their photographs; and K. Harness for providing the Dawson Lake confirmation and photographs.

Specimens examined. — ISUC 703, Dawson Lake, McLean Co., IL, 5 May 1966, J. A. Holman; UIMNH 4962 Normal, McLean Co., IL [no date or collector given]. Color pho¬ tos of the snakes (showing key characters) from the Dawson Lake, Funks Grove and Heyworth localities were deposited with the Illinois Natural History Survey and Illinois State University Collection. Museum abbreviations: BMNH British Museum (Natural History); ISUC Illinois State University Collections; UIMNH University of Illinois Natu¬ ral History Museum.

LITERATURE CITED

Anton, T.G., D. Mauger, C.A. Phillips, M.J. Drezlik, J.E. Petzing, A.R. Kuhns, and J.M. Mui. 2003. Clonophis kirtlandii (Kirtland’s snake). Aggregating behavior and site fidelity. Herpetol. Rev. 34:248-249.

Bavetz, M. 1993. Geographic variation, distribution, and status of Kirtland’s snake, Clonophis kirtlandii (Kennicott) in Illinois. M.S. Thesis, Southern Ill. Univ. Carbondale.

Bavetz, M. 1994. Geographic variation, status, and distribution of Kirtland’s snake ( Clonophis kirtlandii Kennicott) [sic] in Illinois. Trans. Ill. St. Acad. Sci. 87:151-163.

105

Brown, L.E. 1987. A newly discovered population of Kirtland’s snake with comments on habitat and rarity in central Illinois. Bull. Chicago Herpetol. Soc. 22:32-33.

Brown, L.E., R.S. Funk, D. Moll, and J.K. Tucker. 1975. Distributional notes on reptiles in Illinois. Herpetol. Rev. 6:78-79.

Conant, R. 1943. Studies on North American water snakes-1 Natrix kirtlandii (Kennicott). Am. Midi. Nat. 29:313-341.

Conant, R. 1958. A Field Guide to Reptiles and Amphibians of the United States and Canada East of the 100th Meridian. Houghton Mifflin Co., Boston.

Conant, R. and J.T. Collins. 1998. A Field Guide to Reptiles & Amphibians Eastern and Central North America. 3rd ed. Houghton Mifflin Co., Boston and New York.

Dodd, C.K., Jr., G.E. Drewry, R.M. Nowak, J.M. Sheppard, and J.D. Williams. 1985. Endangered and threatened wildlife and plants; review of vertebrate wildlife; notice of review. Part III. U.S. Dept. Interior, Fish and Wildlife Serv., Federal Register 50:37958-37967.

Ernst, C.H. and E.M. Ernst. 2003. Snakes of the United States and Canada. Smithsonian Books, Washington, D.C. and London.

Frymire, D. and A.F. Scott. 2012. Clonophis kirtlandii (Kirtland’s Snake). Herpetol. Rev. 43:447. Garman, H. 1892. A synopsis of the reptiles and amphibians of Illinois. Bull. Ill. St. Lab. Nat. Hist. Vol. Ill, Art. XIII, pp. 215-389.

Gibbons, J.W., D.E. Scott, T.J. Ryan, K.A. Buhlmann, T.D. Tuberville, B.S. Metts, J.L. Greene, T. Mills, Y. Leiden, S. Poppy, and C.T. Winnie. 2000. The global decline of reptiles, deja vu amphibians. BioScience 50:653-666.

Gibson, J. and B. Kingsbury. 2004. Conservation assessment for Kirtland’s snake ( Clonophis kirtlandii). U.S.D.A. Forest Service, Eastern Region, Milwaukee, 29 pp.

Holman, J.A. 1966. Herpetological records from northcentral Illinois. Trans. Ill. St. Acad. Sci. 59:298-300.

Kennicott, R. 1856. Description of a new snake from Illinois. Proc. Acad. Nat. Sci. Philadelphia 8:95-96.

Minton, S., Jr. 1944. Introduction to the study of the reptiles of Indiana. Am. Midi. Nat. 32:438- 477.

Phillips, C.A., R.A. Brandon, and E.O. Moll. 1999. Field Guide to Amphibians and Reptiles of Illinois. Manual 8. Ill. Nat. Hist. Surv., Champaign.

Thurow, G.R. 1993. Clonophis kirtlandii (Kirtland’s snake). Diet. Herpetol. Rev. 24:34-35. Transeau, E.N. 1935. The prairie peninsula. Ecology 16:423-437.

Tucker, J.K. 1994. A laboratory investigation of fossorial behavior in Kirtland’s snake, Clonophis kirtlandii (Kennicott) (Serpentes: Colubridae), with some comments on management of the spe¬ cies. Bull. Chicago Herpetol. Soc. 29:93-94.

106

Transactions of the Illinois State Academy of Science (2012) Volume 105,#3&4 pp. 107-117

received 12/30/1 1 accepted 7/26/12

The Role of Oil Content and Size in Seed Selection by Wild Birds

Kathleen A. Collins and David Joseph Horn1 'Department of Biology, Millikin University 1 184 W. Main St., Decatur, IL 62522, USA

ABSTRACT

We determined whether birds actively select specific varieties of a single seed species, whether seed selection is based on oil content, how birds can distinguish seeds based on oil content, and if seed preferences are equivalent among species. Three trials were con¬ ducted from winter 2007-summer 2007 at Rock Springs Environmental Center in Deca¬ tur, IL. During each trial, seed consumption and bird abundance of each species at each seed variety of black-oil sunflower ( Helianthus annuus ) were recorded. In all trials, seed consumption of the highest oil content seed was 31-49% greater than any other seed vari¬ ety. House Finch ( Carpodacus mexicanus ), American Goldfinch ( Spinus tristis ), and House Sparrow ( Passer domesticus ) preferred seeds with a higher oil content when seed size was equivalent, but selected shorter, deeper seeds with a lower oil content at equiva¬ lent levels to larger seeds with a higher oil content when both varieties were present. Knowing the factors influencing seed choice allows for the development of seed varieties that will be more attractive to songbirds.

INTRODUCTION

Previous studies have examined seed species preferences of wild birds that use feeders (Geis 1980, Horn et al. 2002). Few studies have investigated whether birds choose among multiple varieties of the same seed species, and why birds prefer the seeds of the variety they select. Factors influencing seed choice may include nutritional content of the food (Schaefer et al. 2003), seed size (Willson 1972), bill size (Hespendeide 1966), and behav¬ ior of the bird (Foster 2008). In a study of seed preferences of Henslow’s Sparrows (. Ammodramus henslowii ), two of the preferred seed species had higher energy content, while two of the avoided seed species had the lowest content, indicating that nutrition could play a role in seed selection (DiMiceli et al. 2007). Birds have also selected seed species based on the handling time of a particular species, which in turn, was dependent on the size and shape of the bill (Hrabar and Perrin 2002). When selecting among seeds of a single species, Black-capped Chickadees ( Poecile atricapillus ) and Red-breasted Nuthatches (Sitta canadensis ) selected sunflower seeds based on weight (Heinrich et al. 1997). Weighing sunflower seeds could be beneficial to birds in order to minimize the energy costs of foraging while maximizing the reward (Lima 1985).

We studied four questions associated with why wild birds select specific seeds: (1) Do birds actively select varieties of the same seed species? (2) Do birds select seeds based on nutritional content? (3) Is nutritional content related to seed size? (4) Are there differ¬ ences among species in regards to seed preference? We hypothesized that birds will

108

distinguish among seed varieties of the same species, bird abundance would be greater at seed varieties with greater oil content, and that seed morphology is related to oil content.

In 2006, over 55 million Americans over the age 16 spent more than $4.1 billion on bird food, feeders, and other products (U.S. Fish and Wildlife Service 2007, 2008). Conduct¬ ing studies on whether birds prefer certain seed varieties over others may lead to a better bird feeding experience if preferred varieties are introduced to bird seed blends. In turn, attracting a greater diversity of birds to one’s yard may elevate awareness for the natural world, increase participation in outdoor recreation, and motivate individuals to become involved in conservation issues at a time when participation in some nature-based activi¬ ties is declining (Pergams and Zaradic 2008, U.S. Fish and Wildlife Service 2009).

METHODS

Procedures

Our study consisted of three trials conducted from winter-summer 2007 at Rock Springs Environmental Center in Decatur, Illinois. Tubular bird feeders were utilized (Droll Yan¬ kees B-7R), and there were three or four feeders used, each filled with a different variety of the same species of black-oil sunflower seed, depending on the trial. Black-oil sun¬ flower was selected as it is one of the most preferred seed types among species that com¬ monly use feeders (Geis 1980, Horn et al. 2002). Feeders were hung from a cable approximately 3 m off the ground, and were placed approximately 1 m apart from each other in a straight line. Feeders were rotated in a systematic manner on a weekly basis to ensure that birds were selecting the seed variety not feeder location (Horn et al. 2003), and the initial location of each seed variety within a feeder was determined randomly. Feeder observations took place while seated on a bench looking through a large viewing window. The feeders were approximately 3 m from the window.

To determine whether birds select seeds based on nutritional content and whether there are differences among species in regards to seed preferences, three trials were conducted. Trial 1 examined how seed consumption and relative abundance of birds differed among bird feeders filled with three varieties of small black-oil sunflower seeds (where small seeds are defined as seeds that fit through a 0.5 cm hole [seeds were provided by and sorted at D&D Commodities Ltd., Greeley, CO, USA]). Three oil contents were used: 40.2, 43.6, and 51.4% (oil contents were determined at Eurofins Scientific Inc., Des Moines, IA, USA). Trial 1 ran from 26 Feb -6 Apr. 2007.

Trial 2 examined seed consumption and bird abundance at feeders filled with large black- oil sunflower seeds of varying oil contents (where large seeds are defined as seeds that cannot fit through a 0.5 cm hole). Three oil contents were used: 37.8, 41.1, and 48.1%. The trial ran from 6 Apr -18 May 2007.

Trial 3 examined bird use of sunflower seeds of varying size and oil content. Four seed types were used: small 43.6, large 41.1, small 51.4, and large 48.1%. The trial ran from 18 Jun.-9 Aug. 2007.

Consumption of each seed variety was recorded as the difference in total mass of the seed at the beginning and end of each trial. For each trial, bird abundance was determined

109

through monitoring sessions. Specifically, the number of each bird species present at each feeder was recorded every 5-min during 60-min monitoring sessions, and the means for each session were calculated. Approximately 3 monitoring sessions took place each week with 18, 15, and 24 monitoring sessions being conducted for Trials 1, 2, and 3, respec¬ tively. Observations took place from 08:00-17:00. Because relationships between bird abundance and preference, and seed consumption and preference, are similar (Horn 1995), measures of both bird abundance and seed consumption were used as indicators of preference for each trial.

To determine if nutritional content is related to seed size, we took multiple measures of each seed variety. We measured the depth, length, width, and weight of 75 seeds of each seed variety using digimatic calipers (Mitutoyo CD-6” CSX) and an analytical balance (Mettler AJ100).

To examine whether birds actively select varieties of the same seed species, we con¬ ducted another study concurrent with Trial 3. The study involved observing seed choice of individual birds immediately after feeders were rotated (Fridays) compared to seed choice after birds were adjusted to the feeders’ new positions (Mondays and Thursdays), allowing us to determine if birds change the feeder they visit when the preferred seed variety is moved to a different feeder location. Recordings of seed choice by individual birds were made every 30 sec in 45-min sessions performed every Friday, Monday, and Thursday. To determine which bird to observe, six species were placed into a priority list in random order, and the species of the highest priority present in the feeder area was selected to be observed during the 30-sec interval. Upon selecting a bird to monitor, the first seed choice of the individual was recorded.

For our study, we did not put color bands on birds, and it is possible that some birds were double counted. The number of birds frequenting a feeding station, however, can be very large. Geis and Pomeroy (1993) captured birds in a single yard using mist nets and funnel traps within a lightly developed area in Clarksville, Maryland. In the summer of 1989, they established population estimates ranging from 14 for Tufted Titmouse ( Baeolophus bicolor ) to 5,798 for House Finch ( Carpodacus mexicanus ) with an estimated population size of all feeder birds combined of 6,937 individuals. In our study, the mean number of bird visits per monitoring session in Trial 3 (conducted during summer) was 151 . Thus, if Geis and Pomeroy’s estimate of 6,937 individuals was representative of our study site, a minimum of 46 monitoring sessions would need to be conducted each trial to record each individual within the population.

Statistical analysis

Only species with > 30 observations per trial were used in analyses. To determine whether seed choice was equivalent among the four seed types before and after feeders were rotated, chi-square tests were performed to compare the frequency each seed variety was selected by individual birds immediately after feeders were rotated (Fridays) com¬ pared to the period when birds had time to adjust to the new positions (Mondays and Thursdays combined). Chi-square tests were used because our response variable was the number of times individual birds selected a particular seed variety first as opposed to a response variable such as the mean number of bird visits to a seed variety immediately after rotation compared to 3-5 days after rotation. P-values < 0.05 were considered

110

significantly different. To determine whether bird abundance of each species, total num¬ ber of individuals of all species combined, and species richness per 60-min monitoring session differed among seed types, 95% confidence intervals (CIs) were calculated (Di Stefano 2004), as CIs provide both measures of uncertainty and effect size among treat¬ ments (where the seed varieties are the treatments, Johnson 1999). CIs were also calcu¬ lated to identify differences in seed shape and weight among seed varieties. Treatments with non-overlapping CIs were considered significantly different.

RESULTS

Do birds actively select varieties of the same seed species?

During our examination of whether birds actively select seed varieties (performed concurrently with Trial 3), six species had > 30 observations. All six species, Black- capped Chickadee (N = 83, X2- 166.4, df = 3, P < 0.001), Tufted Titmouse ( Baeolophus bicolor , N = 38, X2 - 45.7, P < 0.001), White-breasted Nuthatch (Sitta carolinensis, N = 48, X2= 11.6, P = 0.009), House Finch (N = 106, X2= 19.4, P < 0.001), American Gold¬ finch (N = 96, X2= 35.8, P < 0.001), and House Sparrow (N = 49, X2 = 62.1 , P < 0.001), had differences in the frequency of visits to a seed variety after adjusting to the seed vari¬ ety being in a new position (Table 1 , see Fig. 1 as a representative example).

Table 1. For six bird species, frequency of seed choice among four varieties of black-oil sunflower (Small - 43.6, Large - 41.1, Small - 51.4, and Large - 48.1) differed between when feeders were first rotated (Friday) compared to after rotation (Monday and Thursday).

Species	Small Fa	-43.6 M/T	Large F	-41.1 M/T	Small F	-51.4 M/T	Large F	-48.1 M/T
amgob	0.19	0.25	0.22	0.06	0.52	0.62	0.07	0.07
BCCH	0.39	0.07	0.26	0.22	0.13	0.22	0.22	0.50
HOFI	0.19	0.28	0.27	0.13	0.35	0.42	0.19	0.17
HOSP	0.14	0.34	0.14	0.23	0.43	0.34	0.29	0.09
TUTI	0.44	0.21	0.00	0.10	0.11	0.28	0.44	0.41
WBNU	0.07	0.11	0.21	0.11	0.21	0.26	0.52	0.53

A F = Friday, M/T = Monday/Thursday.

bAMGO = American Goldfinch, BCCH = Black-capped Chickadee, HOFI = House Finch, HOSP = House Sparrow, TUTI = Tufted Titmouse, and WBNU = White-breasted Nuthatch.

Do birds select seeds based on nutritional content?

Seed consumption was used to determine whether birds select seeds based on oil content. During Trial 1, the greatest consumption of small seed occurred at the highest oil content seed (51.4%) with consumption of the 51.4% oil content seed at 21 kg, 40.2 at 16 kg, and 43.6 at 13 kg. During Trial 2, the greatest consumption also occurred at the seed with the highest oil content (48.1%) with 20 kg consumed followed by 41.1 at 14 kg, and 37.8 at 11 kg.

1 1 1

During Trial 1, no significant differences in bird abundance were observed among the three small-seed varieties with five species having > 30 observations: Downy Wood¬ pecker ( Picoides pubescens , N = 106), Black-capped Chickadee (TV = 63), House Finch (N = 81), American Goldfinch ( N =691), and House Sparrow (N = 45).

No significant differences in bird abundance were observed during Trial 2 among the three large-seed varieties with three species being observed >30 times: Rose-breasted Grosbeak ( Pheucticus ludovicianus , N = 61), House Finch (. N = 48), and American Gold¬ finch ( N - 197).

100

90

80

Small - 43.6 Large -41.1 Small - 5 1 .4 Large - 48.1

Seed type

Figure 1. House Finch seed choice differed among four varieties of black-oil sunflower that varied in size and % oil content when feeders were first rotated (Friday) compared to after rotation (Monday and Thursday, N - 106, X2 = 19.4, df = 3, P < 0.001). Changes in the frequency of seed choice (proportion of total bird visits made to each of the four seed varieties) indicate that birds actively choose specific varieties of seed based on size and oil content.

Is nutritional content related to seed size?

Birds may be able to assess the oil content of a seed based on the seed’s depth, length, and weight, all of which differed in a predictable manner among seed varieties. Collec¬ tively, deep, short, heavy seeds may be cues that birds use to identify high oil content. For example, the depth of small seeds was greatest at the highest oil content seed (Fig. 2a). The length of large seeds, as well as small seeds, decreased with oil content (Fig. 2b). The weight of small, and large seeds, increased with oil content (Fig. 2c), with large seeds being heavier than small seeds regardless of oil content (Fig 2d).

112

Figure 2. 95% confidence intervals of the depth, length, and weight of black-oil sun¬ flower seed varieties that vary in size and % oil content.

Figure 2A. The depth of small seeds was greatest for the highest oil content variety.

Figure 2B. The length of

large seeds, as well as small seeds, decreased as oil content increased.

Figure 2C. The weight of small seeds, as well as large seeds, was greatest for the highest oil content variety.

Sf ®e4 type

Figure 2D. Large seeds were heavier than small seeds, regardless of oil content.

Seed type

Seed type

Are there differences among species in regards to seed preference?

Trial 3 examined seed consumption and bird abundance in relation to both seed size and oil content. The greatest consumption occurred at the highest oil content seed (small 51 .4%) at 52 kg. The small seed with the lower oil content (43.6%) was the second most consumed seed at 35 kg followed by large 48.1% at 29 kg and large 41.1 at 26 kg. Thus, while previous trials found that the higher the oil content, the greater the seed consump¬ tion; in some cases smaller seeds with lower oil content are preferred over larger seeds with higher oil content.

Small seeds were preferred over larger seeds as 43.6% and 51.4 small seeds had a greater abundance of birds of all species combined than 41.1% and 48.1 large seeds. At Rock Springs, 95% CIs of the mean number of birds per 60-min observation session ranged from 1.43-1.96 at small 43.6% seeds, 0.80-1.35 at large 41.1 seeds, 1.95-2.78 at small 51 .4 seeds, and 0.77-1 .20 at large 48.1 seeds.

The greater number of individuals of all species combined at small seed varieties is likely a result of differences among species in regards to seed preference. We found that species that sit and eat at a feeder would select smaller seeds with a lower oil content at an equivalent level to larger seeds with a higher oil content. House Finch ( N = 1031), American Goldfinch ( N = 536), and House Sparrow (A = 155) had their greatest abun¬ dance at small seeds, and had significantly greater abundances at the small 51.4% seed compared to the larger seeds (Fig. 3). 95% CIs of the mean number of House Finch per 60-min observation session ranged from 0.69-1.29 at small 43.6% seeds, 0.35-0.80 at large 41.1 seeds, 0.85-1.63 at small 51.4 seeds, and 0.31-0.69 at large 48.1 seeds. 95% CIs of the mean number of American Goldfinch per 60-min observation session ranged from 0.30-0.52 at small 43.6% seeds, 0.22-0.40 at large 41.1 seeds, 0.64-0.90 at small 51.4 seeds, and 0.14-0.32 at large 48.1 seeds. 95% CIs of the mean number of House Sparrow per 60-min observation session ranged from 0.07-0.23 at small 43.6% seeds, 0.03-0.11 at large 41.1 seeds, 0.12-0.31 at small 51.4 seeds, and 0.02-0.11 at large 48.1 seeds. Downy Woodpecker ( N = 75) abundance was equivalent at each seed variety.

DISCUSSION

We found that wild birds actively choose specific varieties of a single seed, and make choices based on oil content and seed size. In addition, we found that seed characteristics can be used as an indicator of the nutritional content of the seed. Several studies have addressed seed preferences among bird species. A majority of the studies have focused on seed size and shape of multiple seed species (Hespenheide 1966, Willson 1971, Willson 1972, Pulliam 1985). These studies found that birds develop preferences toward certain seed species based on feeding efficiency, or handling time, and the relationship between seed size and bill size. In general, larger-billed birds have the ability to eat larger seeds more effectively than small-billed birds (Hespenheide 1966, Diaz 1990). Studies of a bird’s ability to assess the quality of different varieties of the same seed species based on its physical characteristics are fewer, but have been demonstrated in Black-capped Chickadees (Templeton 2011). Specifically, chickadees were able to select seed heads of spotted knapweed ( Centaurea maculosa) that had greater amounts of gall fly (genus Urophora) larvae using seed head size as one indicator of gall fly abundance.

114

We found that wild birds showed a preference for black-oil sunflower varieties with the highest oil content. Previous studies found that wild birds display preferences for specific micronutrients, and have the ability to distinguish between varying concentrations of sug¬ ars and lipids (McWillams et al. 2002, Schaefer et al. 2003, Pierce et al. 2004, Brown et al. 2010). In a study of four tanager species, Schaefer et al. (2003) concluded that birds were able to distinguish 1% differences in sugar concentrations and 2% differences in lipid concentrations, and typically selected foods with the highest concentrations of each. We found that seed-eating birds were able to distinguish differences in oil content of at least 10%.

Figure 3. American Goldfinch preferred the small black-oil sunflower seeds with 51.4% oil over the three other varieties of black-oil sunflower that varied in size and/or % oil content. An equivalent number of goldfinches were found at small 43.6% seeds compared to large 48.1% seeds indicating that oil content is not the only factor influencing seed choice.

Seed size serves as both a cue for the oil content of a seed and a factor influencing seed choice. We found oil content of sunflower seeds increased with decreasing length, and increasing depth and weight. Birds “weigh” seeds before making a selection (Heinrich et al. 1997). By weighing seeds, birds may not only be maximizing energy costs of foraging (e.g., Lima 1985), but are also able to assess the seed’s nutritional content. We also found that birds used seed size as a factor in seed selection, as some species consumed smaller seeds with less oil content over larger seeds with greater oil content when given a choice. In previous studies, Java Sparrow ( Padda oryzivora) selected safflower seeds that were deeper (Van der Meij and Bout 2000), and Northern Cardinal selected sunflower seeds that were shorter, but thinner (Willson 1972).

1 15

We found differences among species in the seed varieties they choose. Species that sit at a feeder and eat, such as House Finch, American Goldfinch, and House Sparrow selected smaller seeds at equivalent rates to larger seeds, even if larger seeds had higher oil con¬ tents. Thus, for birds that eat food where it is found, ease in handling seeds may be a more significant factor (Willson 1972) than for species that take a seed and consume it elsewhere, where maximizing energy gained from each seed may be a more important factor in seed choice (Lima 1985). This may explain why Black-capped Chickadee, Tufted Titmouse, and White-breasted Nuthatch had the greatest frequency of visits at large, high oil content seeds after adjusting to feeder rotation during the study conducted in conjunction with Trial 3.

Knowledge of seed preferences of birds, and the factors that influence preferences, may ultimately result in improved seed varieties and an enhanced experience for people who feed birds. Specifically, packagers of bird seed could select sunflower varieties of certain size and oil content to create specialized blends based on the birds individuals want to attract. Blends used to attract birds that sit and eat at a feeder might include smaller sun¬ flower seeds, while blends for attracting birds that grab a seed and eat elsewhere might be comprised of larger, higher oil content seeds.

Farmers growing black-oil sunflower face a dilemma. While the seeds are developing in the plant in the field, seed varieties that are bird resistant, particularly to Red-winged Blackbirds ( Agelaius phoenicus ) should be planted (Gross and Hanzel 1991, Bullard 1988, Linz et al. 201 1). However, the seed traits selected in bird-resistant varieties during the 1980’s, including fibrous hulls with high levels of anthocyanins, also created seeds with low oil content and yield (Linz et al. 201 1). While these varieties were less attractive to pest birds (Bullard et al. 1989), they may also be unattractive to birds at feeders that prefer high oil content seed. Future research should examine whether lower oil content black-oil sunflower with fibrous hulls would be selected over other seed species found in bird food blends. In addition, studies should examine if birds have different preferences for certain seed varieties during different seasons. For example, do birds consume even more of the higher oil content seeds in the fall and winter in order to meet the demands of their changing metabolic rates and increase in lipid and protein storage (Karasov and Pinshow 1998, Linkes et al. 2002, McWilliams et al. 2002)?

ACKNOWLEDGMENTS

Funding was provided by D&D Commodities Ltd., Wild Bird Centers of America Inc., and Millikin University. We thank M. O. Christ, M. Evans, D. Hebert, and S. M. Johan¬ sen for assistance in collecting data, and the Macon County Conservation District for allowing us to conduct the study on their property. D. Hebert, S. M. Johansen, L. Lundstrom, G. H. Petrides Sr., and T. E. Wilcoxen reviewed the manuscript before submission.

LITERATURE CITED

Brown, M., C. T. Downs, and S. D. Johnson. 2010. Sugar preferences of a generalist nonpasserine flower visitor, the African Speckled Mousebird ( Colius striatus). Auk 127:781-786.

116

Bullard, R. W. 1988. Characteristics of bird-resistance in agricultural crops. Proceedings of the Vertebrate Pest Conference 13:305-309.

Bullard, R. W., P. P. Woronecki, R. A. Dolbeer, and J. R. Mason. 1989. Biochemical and morphological characteristics in maturing achenes from purple-hulled and oilseed sunflower cultivars. Journal of Agricultural and Food Chemistry 37:886-890.

Diaz, M. 1990. Interspecific patterns of seed selection among granivorous passerines: effects of seed size, seed nutritive value and bird morphology. Ibis 132:467-476.

DiMiceli, J. K., P. C. Stouffer, E. I. Johnson, C. Leonardi, and E. B. Moser. 2007. Seed preferences of wintering Henslow’s Sparrows. Condor 109:595-604.

Di Stefano, J. 2004. A confidence interval approach to data analysis. Forest Ecology and Manage¬ ment 187:173-183.

Foster, M. S. 2008. Freeze-frame fruit selection by birds. Wilson Journal of Ornithology 120:901- 905.

Geis, A. D. 1980. Relative attractiveness of different foods at wild bird feeders. U.S. Department of Interior, Fish and Wildlife Service Special Scientific Report 233. Washington, D.C.

Gross, J. F. and J. J. Hanzel. 1991. Stability of morphological traits conferring bird resistance to sunflower across different environments. Crop Science 31:997-1000.

Heinrich, B., C. C. Joerg, S. S. Madden, and E. W. Sanders Jr. 1997. Black-capped Chickadees and Red-breasted Nuthatches “weigh” sunflower seeds. Auk 114:298-299.

Hespenheide, H. A. 1966. The selection of seed size by finches. Wilson Bulletin 78:191-197.

Horn, D. J. 1995. Perching orientation affects number of feeding attempts and seed consumption by the American Goldfinch ( Carduelis tristis ). Ohio Journal of Science 95:292-293.

Horn, D. J., S. E. Fairbairn, and R. J. Hollis. 2002. Factors influencing the occurrence of birds that use feeders in Iowa. Journal of the Iowa Academy of Science 109:8-18.

Horn, D. J., M. Abdallah, M. K. Bastian, J. R. DeMartini, and R. M. Wilhemi. 2003. Bird abun¬ dance at feeders increases with decreasing distance to cover. Transactions of the Illinois State Academy of Science 96:247-254.

Hrabar, H. D. N. K. and M. Perrin. 2002. The effect of bill structure on seed selection by granivo¬ rous birds. African Zoology 37:67-80.

Johnson, D. H. 1999. The insignificance of statistical significance testing. Journal of Wildlife Management 63:763-772.

Karasov, W. H. and B. Pinshow. 1998. Changes in lean mass and in organs of nutrient assimilation in a long-distance migrant at a springtime stopover site. Physiological Zoology 71:435-448.

Lima, S. L. 1985. Maximizing feeding efficiency and minimizing time exposed to predators: a trade-off in the Black-capped Chickadee. Oecologia 66:60-67.

Linkes, E. T., S. M. Scott, and D. L. Swanson. 2002. Seasonal acclimatization in the American Goldfinch revisited: to what extent do metabolic rates vary seasonally? Condor 104:548-557.

Linz, G. M., J. H. Homan, S. J. Werner, H. M. Hagy, and W. J. Bleier. 2011. Assessment of bird- management strategies to protect sunflowers. Bioscience 61:960-970.

McWilliams, S. R., S. B. Kearney, and W. H. Karasov. 2002. Diet preferences of warblers for spe¬ cific fatty acids in relation to nutritional requirements and digestive capabilities. Journal of Avian Biology 33:167-174.

Pergams, O. R. W. and P. A. Zaradic. 2008. Evidence for a fundamental and pervasive shift away from nature-based recreation. Proceedings of the National Academy of Sciences 105:2295-2300.

Pierce, B. J., S. R. McWilliams, A. R. Place, and M. A. Huguenin. 2004. Diet preferences for spe¬ cific fatty acids and their effect on composition of fat reserves in migratory Red-eyed Vireos ( Vireo olivaceous). Comparative Biochemistry and Physiology, Part A 138:503-514.

Pulliam, H. R. 1985. Foraging efficiency, resource partitioning, and coexistence of sparrow species. Ecology 66: 1 829- 1 836.

Schaefer, H. M., V. Schmidt, and F. Bairlein. 2003. Discrimination abilities for nutrients: which difference matters for choosy birds and why? Animal Behaviour 65:531-541.

Templeton, C. N. 2011. Black-capped Chickadees select spotted knapweed seedheads with high densities of gall fly larvae. Condor 1 13:395-399.

U.S. Fish and Wildlife Service. 2007. 2006 national survey of fishing, hunting, and wildlife-associ¬ ated recreation. National overview.

U.S. Fish and Wildlife Service. 2008. Wildlife watching in the U.S.: The economic impacts on national and state economies in 2006. Addendum to the 2006 national survey of fishing, hunting, and wildlife-associated recreation.

U.S. Fish and Wildlife Service. 2009. Wildlife watching trends: 1991-2006 a reference report. Addendum to the 2006 national survey of fishing, hunting, and wildlife-associated recreation.

Van der Meij, M. A. A. and R. G. Bout. 2000. Seed selection in the Java Sparrow ( Padda oryzi- vora ): preference and mechanical constraint. Canadian Journal of Zoology 78:1668-1673.

Willson, M. F. 1971. Seed selection in some North American finches. Condor 73:415-429.

Willson, M. F. 1972. Seed size preference in finches. Wilson Bulletin 84:449-455.

'

Transactions of the Illinois State Academy of Science (2012) Volume 105,#3&4 pp. 119-125

received 8/8/1 1 accepted 7/26/12

Comparison of Methods to Detect Mesocarnivores in Southern Illinois

• 1 2 1 Clayton K. Nielsen ’ and Susan E. Cooper

'Cooperative Wildlife Research Laboratory and department of Forestry

Southern Illinois University, Carbondale, IL 62901-6504

ABSTRACT

We used trapping, track plates, and remote cameras to survey the distribution of gray foxes ( Urocyon cinereoargenteus ), coyotes ( Canis latrans ), and bobcats ( Lynx rufus) in southern Illinois during 2005-07. Gray fox detection rates were low for all survey tech¬ niques when compared to more abundant mesocarnivore species. We captured 9 gray foxes and 21 bobcats in 7,729 trap-nights and 6 coyotes in 1,416 trap-nights. Track plates (n = 883 survey-nights) resulted in the detection of 6 gray foxes, 1 bobcat, and 1 coyote. Cameras (n = 953 survey-nights) resulted in the detection of 4 gray foxes, 5 bobcats, and 4 coyotes. Although the relative effectiveness of detection methods varied by species, our data are generally in agreement with other survey methods that indicate the relative scar¬ city of gray foxes compared to bobcats and coyotes in southern Illinois.

Key words: bobcat, Canis latrans , coyote, gray fox, Lynx rufus , remote camera, survey, track plate, southern Illinois, trapping, Urocyon cinereoargenteus.

INTRODUCTION

Mesocarnivores often use large home ranges and engage in secretive behavior; these attributes complicate assessments of their abundance and distribution. Three sympatric mesocarnivore species associated with forest cover in southern Illinois are the gray fox ( Urocyon cinereoargenteus ), coyote ( Canis latrans ), and bobcat (Lynx rufus), and wild¬ life biologists have expended many resources monitoring their populations during the past 20 years (e.g., Nielsen and Woolf 2002 a,b). The most reliable estimate of mesocarni¬ vore population trends in Illinois is the Archery Deer Hunter Survey (ADHS), which each year asks hunters to document harvest effort and wildlife sightings. Since its inception in 1991, the ADHS has quantified a 75% decrease in gray fox sightings (Bluett 2007) versus a 15% increase in coyotes and an almost 600% increase in bobcats (Figure 1). Wildlife biologists are unsure why gray fox populations are declining in Illinois, but important limiting factors may include intraguild predation, competition, and mortality from disease (Cypher 2003).

We used traps, track plates (Drennan and Dodd 1998, Olson et al. 2003) and remote cam¬ eras (Silveira et al. 2003, York et al. 2003) to survey for gray foxes, coyotes, and bobcats in southern Illinois, and compared detection rates among survey methods and species. In addition to our assessment of survey methods, we also wished to determine whether field survey methods generally supported mesocarnivore trends observed in the ADHS.

120

Figure 1. Trends in sighting indices for gray foxes, bobcats, and coyotes based on the Illinois Department of Natural Resource’s Archery Deer Hunter Survey, 1996-2006, Illi¬ nois, USA.

STUDY AREA

We conducted field research in 5 southern Illinois counties (Jackson, Johnson, Pope, Union, and Williamson) in the Shawnee National Forest, Crab Orchard National Wildlife Refuge, Giant City State Park, Feme Clyffe State Park, Southern Illinois University Carbondale, City of Carbondale, and on land in private ownership. Elevation in southern Illinois ranges from 91 to 325 m (Netstate 2006) and comprises cropland (39%); upland and bottomland forests (25%) dominated by maples {Acer spp.), oaks {Quercus spp.), and hickory {Carya spp.); and rural grasslands (24%; Luman et al. 1996). Climate in southern Illinois is characterized by 4 distinct seasons with an average annual temperature of 14° C and average annual precipitation of 120 cm (Illinois State Water Survey 2003). Road den¬ sity (1.4 km/km2) and human population density (21.5 persons/km2) are moderately high in this area (Nielsen and Woolf 2002a, &).

METHODS

Trapping

We trapped for gray foxes, coyotes, and bobcats during 3 field seasons: 30 November 2005-18 March 2006, 5 June-4 August 2006, and 9 October 2006-28 February 2007. We used Victor #1.5 padded foothold traps and wire-cage box traps (30 x 30 x 72.5 cm), and also used Woodstream #3 padded foothold traps during winter 2005-06. During the win-

121

ter seasons, we selected trapping areas based on reported animal sightings, evaluation of suitable habitat, historic occurrence, and incidental captures in other studies (Follman 1973, Cypher 1991; C. Nielsen, unpublished data). We used standard dirt-hole and scent- post sets with the foothold traps, and placed box traps under shrubs and concealed them with vegetation. We used a variety of baits including game meat, fatty acid scent discs, commercial scent lures, peanut butter, jelly, and carnivore urine. We checked all traps every morning and immediately released all non-target animals. We immobilized trapped individuals using Telazol® with a dosage rate of 13 mg/kg, or used physical restraint and a blindfold. All animals were marked with individual ear tags. Capture and handling procedures followed Southern Illinois University Carbondale Animal Care and Use Protocol #05-028.

We estimated capture rates for gray foxes, bobcats, and coyotes. Capture rates for gray foxes and bobcats were the total number of individuals captured/ 100 trap-nights; how¬ ever, capture rates for coyotes was based solely on the number of individuals captured in #3.0 footholds/100 trap-nights, as box traps and #1.5 footholds were less likely to capture coyotes.

Track Plates and Remote Cameras

During 5 June-5 September 2006, we surveyed southern Illinois for gray foxes, bobcats, and coyotes using track plates and remote cameras. We divided each county into sections (2.6 km2) and considered sections for surveying if they contained >50% forest and ade¬ quate road access. We used forest cover as a criterion for selecting a section based on knowledge of habitat use by gray foxes, bobcats, and coyotes (Anderson and Lovallo 2003, Bekoff and Gese 2003, Cypher 2003). We included only sections with road access across the section to maximize survey efficiency; 1 17 sections in the 5-county study area met these criteria. From these, we randomly selected approximately 50% of the suitable sections in each county: 19 from Jackson County, 6 from Johnson County, 18 from Pope County, 1 1 from Union County, and 4 from Williamson County.

We placed 5 stations along the main road of each section (Figure 2). The first station was placed on the road in the middle of the section with 2 additional stations placed in each other direction. Each station was separated by 325 m (Conner et al. 1983), and we placed 1 remotely triggered camera and 1 track plate at each station. We randomly selected the first detection device and placed it 50 m from the road, and then placed the second detec¬ tion device 25-50 m from the first. Because it is not uncommon to see gray foxes, bob¬ cats, and coyotes on roads or to find their carcasses near roads (Kolowski and Nielsen 2008), we thought selection of sites near roads would not bias the probability of detecting carnivores. We opportunistically placed detection devices near game trails, creeks, and other probable animal travel ways. We conducted surveys for 4 consecutive nights, weather permitting (Roughton and Sweeny 1982, Nottingham et al. 1989, Engeman and Allen 2000). We recorded the location of all detection devices using a global positioning system unit.

122

Figure 2. Set up of sections (2.6 km2) for track plate and camera detection surveys ran during June-September 2006, southern Illinois, USA. One camera and 1 track plate were placed at each station marked by the white dot. Stations were separated by 325 m.

At each detection device, we randomly selected a lure that should appeal to all mesocamivore species potentially present. We used a commercial food-based lure (Lenon’s Fox #1 Super All Call), a mixture of fish oil and shellfish oil, or a fatty-acid- scent disc. We replaced lures if they had been obviously removed by animals visiting the site. As an attractant, we also sprayed the area around each detection device with carni¬ vore urine. We used Moultrie® 100v2 Gamespy (2.1 megapixel) and Moultrie® 200 Gamespy (3.1 megapixel) cameras. We set cameras to take 2 pictures/min when triggered and attached the camera approximately 0.5 m above the ground on a tree. We then dug a dirt hole approximately 2 m from the camera in which the lure was placed. We down¬ loaded all images from the cameras and identified species.

Track plates were 0.64-m2 treated plywood boards covered with aluminum flashing and sprayed with a 1:4 mixture of dissolved carpenter’s chalk and denatured ethyl alcohol (Drennan and Dodd 1998, Olson et al. 2003). We placed a lure in the center of the track plate and identified carnivore tracks to species. We calculated detection rates using cam-

123

eras and track plates for gray foxes, bobcats, and coyotes based on the number of individuals detected/ 100 survey-nights.

RESULTS

Trapping

We captured 9 gray foxes and 21 bobcats in 7,729 trap-nights and 6 coyotes in 1,416 trap-nights. Capture rates (individuals/100 trap-nights) were 0.12, 0.27, and 0.42, for gray foxes, bobcats, and coyotes, respectively (Table 1). Other animals captured included Vir¬ ginia opossums ( Didelphis virginiana ), raccoons ( Procyon lotor), domestic dogs (C. familiaris), striped skunks (. Mephitis mephitis ), eastern cottontails ( Sylvilagus floridanus ), squirrels ( Sciurus spp.), domestic cats (Felis catus ), an eastern box turtle ( Terrapene Carolina ), and a woodchuck (Marmota monax; Table 2).

Table 1. Detection rates (individuals/100 survey-nights) of sympatric carnivores based on trapping, track plate, and camera surveys conducted during December 2005-February 2007, southern Illinois, USA.

Device (# survey-nights)	Gray fox	Bobcat	Coyote
Box trap (2,163)	0.05	0.37	N/A
Foothold (5,566)	0.14	0.23	0.42a
Track plate (883)	0.68	0.11	0.11
Camera (953)	0.42	0.53	0.42
Totals (9,565)	0.20	0.28	0.34
aCapture rate for coyotes was based on total trap nights using #3.0 foothold traps ( n = 1,416).

Table 2. Capture totals based southern Illinois, USA.	on 7,756 trap-nights during December 2005-	-March 2007,
Species	Footholds	Box trap	Totals
Virginia opossum	104	108	212
Raccoon	89	65 •	154
Striped skunk	9	14	23
Bobcat	13	8	21
Domestic dog	13	0	13
Eastern cottontail	9	0	9
Gray fox	8	1	9
Coyote	6a	0	6
Squirrel	3	0	3
Domestic cat	2	1	3
Woodchuck	1	0	1
Turtle	1	0	1
Totals	258	197	455
aCapture total for coyotes was 1,416).	i based on total trap nights using #3.0 foothold traps (/? =

124

Track Plates and Remote Cameras

Track plates (n = 883 survey-nights) resulted in the detection of 6 gray foxes, 1 bobcat, and 1 coyote (Table 1). Cameras (n = 953 survey-nights) resulted in the detection of 4 gray foxes, 5 bobcats, and 4 coyotes. Other species detected using these methods included raccoons, opossums, domestic dogs, squirrels, eastern cottontails, striped skunks, turkey vultures ( Cathartes aura), black vultures ( Coragyps atratus), a wood¬ chuck, a domestic cat, a wild turkey ( Meleagris gallopavo), and a nine-banded armadillo (Dasypus novemcinctus).

Overall, remote cameras (1.36 detections/ 100 survey-nights) were more effective for detecting these 3 focal carnivore species than track plates (0.91 detections/ 100 survey- nights). Both bobcats and coyotes were more effectively detected by cameras than the other devices, but gray foxes appeared to be best detected by track plates (Table 1).

DISCUSSION

Our trapping data are in agreement with the ADHS, which indicates the relative scarcity of gray foxes in Illinois compared to bobcats and coyotes (Bluett 2007). We captured twice as many bobcats as gray foxes and coyotes, but the capture rate for coyotes (0.42) was higher than for both gray foxes (0.1 1) and bobcats (0.27). We based capture rates for coyotes solely on the #3.0 foothold traps because #1.5 foothold traps and the box traps were too small to effectively catch or hold most coyotes. This bias should not exist for gray foxes and bobcats (Zezulak 1980, Fuller et al. 1995, Gabriel 2006). Using these cap¬ ture rates as coarse indices of relative abundance, gray foxes are scarcer than bobcats or coyotes. However, the species differ in likelihood of capture. For example, coyotes are wary of novel objects, especially within their core home ranges, making them difficult to trap (Sauvajot et al. 2000, Sequin et al. 2003, Mettler and Shivik 2007), whereas gray foxes and bobcats may not possess this trap shyness (Fritzell and Haroldson 1982). Therefore, capture rates from this study actually may underestimate relative abundance of coyotes in southern Illinois.

The relative effectiveness of detection methods used during this study varied by species. Although remote cameras detected the most bobcats and coyotes, cameras did not appear to detect gray foxes as well as did track plates. Again, differential detectability of gray foxes and coyotes may explain some variation in the number of detections obtained by devices such as track plates or cameras. Coyotes are wary of cameras (Sauvajot et al.

2000, Sequin et al. 2003), and it is likely they would have similar reactions to track plates, as concluded by Heske et al. (201 1). Track plate surveys could be improved (e.g., by creating a more natural-looking tracking surface such as sand or soil) to make them more attractive to wary species (Heske et al. 2011).

There are many ways to conduct carnivore surveys (Roughton and Sweeny 1982, Gese

2001, Sargeant et al. 2003, Barea-Azcon et al. 2007). The most efficient surveys would allow researchers to run the fewest number of devices in the smallest area for the fewest nights without risking inefficient sampling (Field et al. 2005, Joseph et al. 2006). Complicating matters is that at different times of the year, animals are more or less likely to visit devices (Gompper et al. 2006). For example, during the breeding and juvenile dispersal seasons, home ranges expand (Follman 1973, Andelt and Gipson 1979, Sawyer

125

and Fendley 1990), whereas home ranges contract during the whelping season (Follman 1973, Sawyer and Fendley 1990). There also may be an increased chance of detection during seasons when food is less plentiful, due to animals roaming more widely in search of food.

Altering survey methods in several ways may have produced higher detection rates of mesocarnivores in southern Illinois. Because coyotes may be more reluctant to visit non¬ natural devices (Sauvajot et al. 2000, Sequin et al. 2003, Mettler and Shivik 2007), dirt- circle scent stations may seem less out of place than track plates. Remote cameras also may appear suspicious to coyotes (Sauvajot et al. 2000, Sequin et al. 2003). Also, we conducted surveys in the summer, at which time the maximum number of gray foxes should be present due to the birth pulse. Juvenile animals may be less wary of survey devices (Windberg and Knowlton 1990). It is also difficult to determine the number of nights to survey for multiple species that have differently-sized home ranges. Because gray foxes inhabit relatively small home ranges when compared to bobcats and coyotes, they should have been more likely to encounter a detection device within the 2.6-km2 study sections, whereas a coyote or bobcat may never enter that section in the 4 nights we ran surveys.

Our initial assessment of mesocarnivore distribution in Illinois has refined objectives for continuing research on mesocarnivores in Illinois. Specifically, we are using remote cam¬ eras (and not the other techniques discussed in this paper) to conduct occupancy surveys (MacKenzie et al. 2006) of gray foxes and sympatric mesocarnivores at 1,118 sites in 357 2.6-km" sections in the 16 southernmost counties of Illinois (C. Nielsen, unpublished data). These surveys will provide a more rigorous assessment of factors affecting occu¬ pancy and distribution of gray foxes, coyotes, and bobcats, focusing primarily on species interactions and habitat characteristics.

ACKNOWLEDGMENTS

We acknowledge the Illinois Department of Natural Resources’ Furbearer Fund for fund¬ ing this research, and B. Bluett of the Illinois Department of Natural Resources for logistical support. We also thank the Cooperative Wildlife Research Laboratory, Depart¬ ments of Zoology and Forestry, Graduate School, and College of Agricultural Sciences at Southern Illinois University Carbondale. E. Hellgren, E. Schauber, and 3 anonymous reviewers provided useful comments on an earlier draft of this work. We recognize C. Effinger, D. Haan, L. Hillard, and A. Nollman for assistance with field work. Finally, we thank private landowners, the Illinois Department of Natural Resources, the U.S. Fish and Wildlife Service, and the U.S. Forest Service for access to properties for surveys.

LITERATURE CITED

Andelt, W. F., and P. S. Gipson. 1979. Home range, activity, and daily movements of coyotes. Jour¬ nal of Wildlife Management 43:944-951.

Anderson, E. M., and M. J. Lovallo. 2003. Bobcat and lynx {Lynx rufus and Lynx canadensis). Pages 758-786 in G. A. Feldhamer, B. C. Thompson, and J. A. Chapman, editors. Wild mam¬ mals of North America: Biology, management, and conservation. John Hopkins University Press, Baltimore, Maryland, USA.

126

Barea-Azcon, J. M., E. Virgos, E. Ballesteros-Duperon, M. Moleon, and M. Chirosa. 2007. Survey¬ ing carnivores at large spatial scales: a comparison of four broad-applied methods. Biodiversity Conservation 16:1213-1230.

Bekoff, M., and E. M. Gese. 2003. Coyote ( Canis latrans). Pages 467-481 in G. A. Feldhamer, B. C. Thompson, and J. A. Chapman, editors. Wild mammals of North America: Biology, manage¬ ment, and conservation. John Hopkins University Press, Baltimore, Maryland, USA.

Bluett, B. 2007. 2006 Archery deer hunter survey. Wildlife Diversity Program Note 06-4. Illinois Department of Natural Resources, Springfield, Illinois, USA.

Conner, M. C., R. F. Labisky, and D. R. Progulske. 1983. Scent-station indices as measures of population abundance for bobcats, raccoons, gray foxes, and opossums. Wildlife Society Bulle¬ tin 1 1: 146-152.

Cypher, B. L. 1991. Coyote foraging dynamics, space use, and activity relative to resource varia¬ tion at Crab Orchard National Wildlife Refuge, Illinois. Dissertation, Southern Illinois Univer¬ sity, Carbondale, Illinois, USA.

Cypher, B. L. 2003. Foxes. Pages 511-546 in G. A. Feldhamer, B. C. Thompson, and J. A. Chap¬ man, editors. Wild mammals of North America: Biology, management, and conservation. John Hopkins University Press, Baltimore, Maryland, USA.

Drennan, P. B., and N. F. Dodd. 1998. Use of track stations to index abundance of sciurids. Journal of Mammalogy 79:352-359.

Engeman, R. M., and F. Allen. 2000. Overview of a passive tracking index for monitoring wild canids and associated species. Integrated Pest Management Reviews 5:197-203.

Field, S. A., A. J. Tyre, and H. P. Possingham. 2005. Optimizing allocation of monitoring effort under economic and observational constraints. Journal of Wildlife Management 69:473-482.

Follman, E. H. 1973. Comparative ecology and behavior of red and gray foxes. Dissertation, South¬ ern Illinois University, Carbondale, Illinois, USA.

Fritzell, E. K., and K. J. Haroldson. 1982. Urocyon cinereoargenteus . Mammalian Species 189:1-8.

Fuller, T. K., S. F. Berendzen, T. A. Decker, and J. E. Cardoza. 1995. Survival and cause-specific mortality rates of adult bobcats {Lynx rufus). American Midland Naturalist 134:404-408.

Gabriel, M. W. 2006. Exposure to Anaplasma phagocytophilum and ticks in gray foxes ( Urocyon cinereoargenteus ) in northern Humboldt County, California. Thesis, Humboldt State University, Humboldt, California, USA.

Gese, E. M. 2001. Monitoring of terrestrial carnivore populations. Pages 372-396 in J. F. Gittle- man, S. M. Funk, D. W. MacDonald, and R. K. Wayne, editors. Carnivore Conservation. Cam¬ bridge University Press and The Zoological Society of Fondon, Cambridge, Great Britain.

Gompper, M. E., R. W. Kays, J. C. Ray, S. D. Fapoint, D. A. Bogan, and J. R. Cryan. 2006. A comparison of noninvasive techniques to survey carnivore communities in northeastern North America. Wildlife Society Bulletin 34:1 142-1151.

Heske, E. J., T. W. Rodges, and T. R. Van Deelen. 2011. Detection bias in noninvasive track sur¬ veys of mammalian predators in Illinois. Transactions of the Illinois Academy of Science 104: 137-143

Illinois State Water Survey, State Climatologist Office. 2003. Climate of Illinois. <http://www .sws.uiuc.edu/atmos/statecli/General/Illinois-climate-narrative.pdf#search=%22annual%20avera ge%20temperatures%20southern%20illinois%22>. Accessed 27 Aug 2006.

Joseph, F. N., S. A. Field, C. Wilcox, and H. P. Possingham. 2006. Presence-absence versus abun¬ dance data for monitoring threatened species. Conservation Biology 20:1679-1687.

Kolowski, J. M., and C. K. Nielsen. 2008. Using Penrose distance to identify potential risk of wild¬ life-vehicle collisions. Biological Conservation 141:1 1 19-1 128.

Fuman, D., M. Joselyn, and F. Suloway. 1996. Critical trends assessment project: landcover data¬ base of Illinois, 1995. Illinois Natural History Survey, Springfield, Illinois, USA.

MacKenzie, D. I., J. D. Nichols, J. A. Royle, K. H. Pollock, F. F. Bailey, and J. E. Hines. 2006. Occupancy estimation and modeling: inferring patterns and dynamics of species occurrence. Academic Press, New York, New York, USA.

Mettler, A. E., and J. A. Shivik. 2007. Dominance and neophobia in coyote ( Canis latrans) breed¬ ing pairs. Animal Behaviour Science 102:85-94.

Netstate. 2006. The geography of Illinois, <http://www.netstate.com/states/geography/il _geography.htm>. Accessed 27 Aug 2006.

127

Nielsen, C. K., and A. Woolf. 2002a. Habitat-relative abundance relationship for bobcats in south¬ ern Illinois. Wildlife Society Bulletin 30:222-230.

Nielsen, C. K., and A. Woolf. 2002 b. Survival of unexploited bobcats in southern Illinois. Journal of Wildlife Management 66:833-838.

Nottingham, B. G., Jr., K. G. Johnson, and M. R. Pelton. 1989. Evaluation of scent-station surveys to monitor raccoon density. Wildlife Society Bulletin 17:29-35.

Olson, T. L., J. S. Dieni, F. G. Lindzey, and S. H. Anderson. 2003. Swift fox detection probability using tracking plate transects in southeast Wyoming. Pages 93-98 in M. A. Sovada and L. Car- byn, editors. The swift fox: ecology and conservation of swift foxes in a changing world. Cana¬ dian Plains Research Center, Regina, Saskatchewan, Canada.

Roughton, R. D., and M. W. Sweeny. 1982. Refinements in scent station methodology for assessing trends in carnivore populations. Journal of Wildlife Management 46:217-229.

Sauvajot, R. M., E. C. York, T. K. Fuller, H. S. Kim, D. A. Kamradt, and R. K. Wayne. 2000. Distribution and status of carnivores in the Santa Monica Mountains, California: preliminary results from radio telemetry and remote camera surveys. Pages 1 13-123 in J. E. Keeley, M. Baer- Keeley, and C. J. Fotheringham, editors. Second interface between ecology and land develop¬ ment in California. U.S. Geological Survey Open-File Report 00-62, Sacramento, California, USA.

Sawyer, D. T., and T. T. Fendley. 1990. Seasonal home range size and movements behavior of the gray fox on the Savannah River site. 1990 Proceedings annual conference of Southeastern Association of Fish and Wildlife Agencies 44:379-389.

Sequin, E. S., M. M. Jaeger, P. F. Brussard, and R. H. Barrett. 2003. Wariness of coyotes to camera traps relative to social status and territory boundaries. Canadian Journal of Zoology 81:2015- 2025.

Silveira, L., A. T. A. Jacomo, J. Alexandre, and F. Diniz-Filho. 2003. Camera trap, line transect census and track surveys: a comparative evaluation. Biological Conservation 1 14:351-355.

Windberg, L. A., and F. F. Knowlton. 1990. Relative vulnerability of coyotes to some capture procedures. Wildlife Society Bulletin 18:282-290.

York, E. C., T. F. Moruzzi, T. K. Fuller, J. F. Organ, R. M. Sauvajot, and R. M. DeGraaf. 2003. Description and evaluation of a remote camera and triggering system to monitor carnivores. Wildlife Society Bulletin 29:1228-1237.

Zezulak, D. S. 1980. Northeastern California bobcat study. California Department of Fish and Game, Nongame Wildlife Investigations, Project E_W_2, Job IV_1.6, Sacramento, California, USA.

Transactions of the Illinois State Academy of Science (2012) Volume 105,#3&4 pp. 129-143

received 6/1/12 accepted 1/14/13

Selection of Parturition Sites by Migrating and Dispersing Female White-tailed Deer

in Illinois

Charles M. Nixon,1,9 Philip C. Mankin,2, 10 Dwayne R. Etter,’ Lonnie P. Hansen,4 Paul A. Brewer,5 James E. Chelsvig,6 Terry L. Esker,7 and Joseph B. Sullivan8 'Illinois Natural History Survey, Champaign, IL 61820 department of Natural Resources and Environmental Sciences, University of Illinois

1 102 South Goodwin, Urbana, IL 61820

3Michigan Department of Natural Resources, 8562 East Stoll Road, Lansing, MI 48823 4Missouri Department of Conservation, 1110 South College Ave., Columbia, MO 65201 Illinois Department of Natural Resources, One Natural Resources Way

Springfield, IL 62702

6Forest Preserve District of Cook County, 536 North Harlem Ave.,

River Forest, IL 60305

7Illinois Department of Natural Resources, 4295 North 1000th St., Newton, IL 62448

8RR #3, Box 201 A, Mt. Sterling, IL 62353 Corresponding author’s present address: 19 Westwood, Monticello, IL 61856

e-mail: cenixon@mchsi.com

10Present address: 5 Steele A Way, Brooksville, ME 04617

ABSTRACT

Emigration behavior by female white-tailed deer ( Odocoileus virginianus ) is an important attribute of population dynamics in Illinois. The factors influencing the process of select¬ ing a new home range are largely unknown, yet may affect hunting success, other interac¬ tions with human activities, and deer social behaviors. Twenty-five radio-marked dispers¬ ing (N = 14) and migrating (N = 11) female white-tailed deer (4 adults, 7 yearlings, 14 fawns when marked) were followed from 3 sites (2 in central and 1 in northern Illinois) to the home ranges selected for parturition and fawn rearing. Dispersing deer (i.e., those making one-way movements to a new home range) moved an average of 44.9 ± SE 6.4 km whereas migrators (i.e., those moving to a new range) moved 14.0 ± SE 7.2 km from their natal ranges. Females moved rapidly in nearly a straight line to their new home range readily crossing roads, rivers, and under high tension power lines on their journey. Females marked in east-central and northern Illinois where forest cover totaled <5% of the landscape, selected sites with greater forest cover, grasslands, forest patch size and density, and total patch richness than were available in random locations. Habitats selected by females in west-central Illinois where more forest cover was available did not differ from random sites. Parturition ranges (used May 1 5— July 15) at all 3 sites con¬ tained fewer houses and roads and more forest patches compared with the natal ranges and the larger new range (used May 15-October 1). Migrating females survived longer than dispersing females, but female life span and fawn survival were not significantly affected by the habitat composition of the new home ranges.

130

INTRODUCTION

Emigration behavior is an important attribute of white-tailed deer of both sexes within the fragmented landscapes that characterize much of the Great Plains and the glaciated por¬ tions of Illinois (VerCauteren and Hygnstrom, 1994; Brinkman etal., 2005; Nixon et al., 2007). Emigration movements usually occur when deer are 10-13 mo old and may involve travel up to 200 km or more from their natal ranges (Sparrowe and Springer, 1970; Gladfelter, 1978; Dusek et al., 1989; Nixon etal., 1991). Emigration by female deer in Illinois has been triggered by mother and sibling movements, orphaning, social position, birth sequence with later born fawns in a social group more likely to emigrate, and, with many fawns breeding, a search for a suitable parturition site (Etter et al., 1995; Nixon et al., 2007; Nixon et al., 2008; Nixon et al., 2010).

Less attention has been devoted to examining the characteristics of home ranges selected by female immigrants or why they stop an emigration and settle on a new range. Possible internal incentives to stop include hunger, fatigue from travel, and changes in reproduc¬ tive hormone levels associated with impending parturition (Plotka et al., 1977; Ozoga et al., 1982; Stamps, 2001; Wiems, 2001; Andreassen et al., 2002). External factors may also affect final selection of a new home range including an immigrant’s attraction to a parturition site if >1 yr old, an attraction to or avoidance of conspecifics, a search for a landscape similar to that of the natal range, and avoidance of large rivers, interstate high¬ ways, and urban centers (Nelson and Mech, 1992; Danchin et al., 2001; Stamps, 2001; Klaver et al., 2008; Long et al., 2010). In Illinois, it is likely the sounds, scents, and sights of human activity in and around various sites that are otherwise adequate in terms of nutrition and cover, may also influence the decision on where to stop an emigration movement.

We hypothesized that selection of a stopping point for an emigration movement was a random event, based on the effect of various external and internal stimuli. Selection of a home range is a second order scale of habitat selection, when emigrating females select a landscape mosaic of forest, grasslands, and crop fields supportive of their immediate needs (Orians and Wittenberger, 1991; Morrison et al., 2006; Godvik et al., 2009; Grovenburg et al., 2010). Such a selection affects future survival, reproductive success, and overall fitness. We examine some of the factors that may influence immigrating Illi¬ nois breeding females in their selection of a new home range in late spring just before the critical period of parturition and fawn rearing.

METHODS

We used 3 study areas (1 in northern Illinois and 2 in central Illinois) to investigate the demographic characteristics, seasonal movements, social behavior, habitat selection, and the importance of refugia for deer in landscapes shaped by human activities (Nixon et al., 1991). Each area included a wooded public park surrounded by privately owned agricul¬ tural fields. The 2,953-ha east-central (EC) study area lies within Piatt County. In 1985, this region was only 2.4% forested, with forests restricted to areas too rough or wet to farm (Iverson et al., 1989). The Sangamon River bisected the study area from NE to SW and provided a forested emigration route for deer. The 5,942-ha west-central area (WC), located in Brown and Adams counties, was more rugged with forests covering about 22%

131

of the landscape (Iverson et al., 1989) and where farms generally included a livestock component. The 1,648-ha northern (NO) area in DeKalb County lies within a region also intensively row cropped with forests occupying about 1.3% of the landscape. Urban sprawl was evident in this region which includes Chicago and Rockford, the largest cities in Illinois. There was no major river corridor associated with either the WC or NO study area.

The 25 female white-tailed deer reported in this paper were part of a larger comprehen¬ sive study that included 286 deer captured during 1980-85 (EC) and 352 captured during 1990-1993 (WC and NO). These deer were captured using rocket-propelled or drop nets (Hawkins et al ., 1968) (Table 1). Deer were manually restrained and marked with ear tags and radio transmitters (Wildlife Materials, Carbondale, IL; Telonics, Inc., Mesa, AZ; or Advanced Telemetry Systems, Inc., Isanti, MN). Our capture and marking methods were approved by the University of Illinois Institutional Animal Care and Use Commit¬ tee.

Table 1. Life history of emigrating female white-tailed deer radio-tracked to parturition sites away from the natal range in Illinois during 1980-1985 and 1990-1993. Contact months represents the period from capture to last contact or death (NO = northern, EC = east-central, WC = west-central study areas).

Tag	Year marked	Age when marked	Type of movement	Radio fixes (N)	Distance moved (km)	Contact (mo)	Life span (mo)	Cause of death
NO A	jrca
155	1991	Fawn	Dispersal	28	58.5	33		Alive1
159	1991	Fawn	Migration	60	22.0	36	67	Auto
212	1992	Fawn	Dispersal	13	51.4	8	28	Auto
178	1992	2 yr	Migration	75	6.4	26		Alive1
196	1992	3 yr	Migration	69	8.5	25	77	Train
EC Area
262	1980	Fawn	Migration	167	6.9	45	53	Bow cripple
276	1980	Fawn	Dispersal	11	57.3	9	17	Gun
218	1983	Fawn	Dispersal	30	25.5	8	17	Gun
507	1983	Fawn	Dispersal	48	65.0	25	29	Gun
577	1984	Fawn	Dispersal	23	36.8	8	17	Gun cripple
636	1985	Fawn	Dispersal	34	13.7	8	17	Gun
286	1980	Yearling	Dispersal	286	20.2	9	30	Gun
325	1981	Yearling	Dispersal	67	52.9	36	54	Auto
338	1981	Yearling	Dispersal	271	12.6	10	30	Gun
542	1984	Yearling	Dispersal	38	31.3	18	41	Unknown
619	1985	Yearling	Dispersal	19	22.8	8	28	Bow
266	1980	Yearling	Migration	157	7.3	44	65	Gun
558	1984	2 yr	Migration	111	3.2	33	65	Gun
238	1980	3 yr	Migration	113	20.8	10	54	Gun cripple
WC Area	' .. MNMiMi
55	1991	Fawn	Dispersal	9	128.0	13		Alive1
117	1991	Fawn	Dispersal	7	60.3	10	17	Gun
138	1991	Fawn	Migration	36	5.0	8	17	Gun cripple
109	1991	Fawn	Migration	102	26.7	35	42	Gun
194	1992	Fawn	Migration	28	17.6	21	66	Gun
115	1991	Yearling	Migration	102	26.7	37	65	Gun

1 Alive at end of study. Cause of death unknown.

132

Deer were aged using tooth replacement and wear from incisors and premolars observed on live deer as either fawn (<12 mo), yearling (12-23 mo), 2 yr old (24-35 mo), or adult (>36 mo) (Severinghaus, 1949). Pregnancy was determined for fawn captures using blood progesterone levels (Wood et al., 1986) or from direct observation of all 25 females on the new range. We captured deer in the order they were lured to bait sites with no random capture selection attempted. Radios were allocated according to their availability each year with a priority given to radio-marking family groups (e.g., females with associated fawns and/or yearlings).

Females were radio tracked using 2 truck-mounted, 8-element yagi antennas aligned in a null configuration. Radio locations were validated using an unpublished program for the Apple II computer (L. P. Hansen, Illinois Natural History Survey) for the EC females and the software program LOCATE II (Nams, 1990) for the WC and NO females. Home range boundaries for each female was calculated for each natal range and for the new range selected by each immigrant using all radio fixes and observations (Table 1) and then superimposed on topographic maps (Lesage et al., 2002).

We defined dispersal as the one-way emigration to a new home range at least 12 km from the natal range. Migration was defined as a movement by females at least 3 km away from the natal range to a new home range where parturition took place. These females then returned to their natal range each winter. Migrating females >2 yr old when first captured were considered to have already located their summer ranges as fawns or year¬ lings. We base this assumption on 2 known characteristics of white-tailed deer in Illinois: first, known female emigrations from our study areas have always begun following their ejection from their mother’s parturition site in late spring at 10-13 mo old or, for various reasons (such as reproductive state, social status, loss of kin) delayed until 21-24 mo old (Nixon et al., 1991 ); second, most mother’s strong attraction each year to a site of parturi¬ tion once a successful birth has occurred (Ozoga et al., 1982; Tierson et al., 1985; Dusek et al., 1989; Nixon et al., 1992; Etter et al., 1995) and verified by the behavior of our marked females followed for longer than a single season (14 of 25 deer).

The new home range settled by female deer was occupied from late February until Octo¬ ber 1, the date when fawns were considered finally weaned (Clutton-Brock et al., 1982). The part of the new home range that was occupied May 15-July 15 was considered the parturition range, and the habitats within this range were analyzed separately from the total new home range. Generally, only migrating females moved as early as late Febru¬ ary, whereas dispersing deer typically started moving in May. Each delineated home range was cover mapped as upland or bottomland forest <40 or >40 yr old, permanent grasslands (CRP lands, pastures active or abandoned, roadsides, cultivated hay fields, mowed grasses of cemeteries, lawns, and proposed housing developments), row crops of corn ( Zea maize ) or soybeans ( Glycine max), and permanent water bodies. Forest oversto¬ ries at all sites were similar to the natal sites when forest age and flooding frequency was also similar. Upland forest >40 yr old were dominated by various oaks ( Quercus sp .) and hickories ( Carya sp.). Those upland forests <40 yr old were mixtures of oaks, hickories, elms ( Ulmus sp.), black walnut (Jug lands niger), honey locust ( Gleditsia triacanthos) , osage orange ( Machura pomifera), and shingle oak ( Quercus imbricaria) . Bottomland forest <40 or >40 yr old that was frequently inundated was nearly a monotype of silver maple (Acer saccharinum ), whereas dryer sites were mixtures of silver maple, hackberry

133

( Celtis occidentalis), sycamore ( Platanus occidentalis), black walnut, cottonwood ( Popu - lus deltoides ), ashes ( Fraxinus sp .), and bur oak ( Quercus macrocarpa).

Data Analysis

Because we used both migrating (N = 11) and dispersing deer (N = 14) for this analysis, we first compared the landscape characteristics of the new home ranges selected by the 2 types of movement behaviors using the t-test. The landscape variables used to analyze landscape use (the natal range, travel corridor used by each deer, parturition range, and new home range) are listed in Table 2. We separated the parturition ranges from the new home ranges because of the importance of cover to early fawn survival (Rohn et al., 2007).

Table 2. Landscape attributes and human presence within the natal ranges, the new home ranges (May 15-October 1), and the parturition ranges (May 1 5— July 15) selected by 25 emigrating female white-tailed deer in Illinois during 1980-1985 and 1990-1993.

Landscape Variables

Travel Corridors

Roads — all state and federal highways Rivers — all river crossings

Forest patches — all distinct forest patches encountered Power lines — all high tension lines carrying at least 34,500 volts Home Ranges

Vegetation stage — dormant or actively growing when emigration began

Female present — females actually seen on the new home ranges

Percent forest — percent of home range forested

Percent cropland — percent of home range in active crops

Percent grasses — percent of home range in permanent grass

Richness — sum of all distinct habitat patches

Nearest neighbor — distance between forest patches (km)

Forest patch density — number of forest patches per 10 ha Average size — average size of forest patches (ha)

Houses — all occupied dwellings in the home range

Forest age — <40 or >40 years

Forest type — upland or bottomland forest

Several of these landscape variables were significantly correlated as determined by the nonparametric Spearman’s Rho test (Zar, 1999), a result expected because the landscape is finite and higher amounts of 1 habitat mean lower amounts of another. For example, negative correlations included percent cropland with percent forest (Rho = -0.82, P < 0.0001), percent grassland (Rho = -0.61 , P = 0.001), and forest patch size (Rho = -0.69, P = 0.0002), while positive correlations included forest patch size with percent forest (Rho = 0.70, P < 0.0001) and percent grassland (Rho = 0.40, P = 0.047) as well as num¬ ber of forest patches with distance between forest patches (Rho = 0.63, P = 0.0007). We used stepwise regression (backward selection) to reduce the number of variables, allow¬ ing the analysis to sort for the appropriate (less correlated) landscape variables. Backward selection regression was chosen because it is better able to identify significant independ-

134

ent variables than is forward selection when there is multi-collinearity among variables (Zar, 1999).

Travel Characteristics

The deviation in travel direction from a straight line was examined by comparing radio locations along each travel route to a straight line drawn from the center of the natal range to the center of the new home range. We also tested for a deviation in the angle of dispersal for 14 deer from the EC area compared to the general direction azimuth of the river corridor. We used the Wilcoxon 2-sample test to examine for differences in the 2 measurements (Zar, 1999).

The initial direction (degrees azimuth) a female moved away from the natal range was measured and placed on USGS topographic maps (published 1971-1984 for EC, 1971- 1980 for NO, 1981 for WC). We then compared this chosen direction with 6 alternative directions that were 30°, 60°, and 90° on either side of this route (and the same length as the actual route) using one-way ANOVA and recorded the number of roads (Illinois Secretary of State highway maps), rivers, forest patches, and high voltage power transmission lines (Illinois Commerce Commission 1971, 1997) encountered along these azimuths. We felt limiting this effort to 90° on either side of the actual route best pro¬ vided an insight into the possible travel choices available to the marked females as they were leaving their natal range. Forest patches were located along each azimuth using topographic maps and rechecked using LANDSAT satellite imagery through the internet program Google Earth.

Recently there has developed interest in the biological effects of the electromagnetic forces associated with high voltage power lines. We were interested in the biological effects, if any, of electromagnetic forces on deer travel routes and on the selection of a new range. Both roe deer ( Capreolus capreolus ) and domestic cattle ( Bos taurus) tend to align their resting body axis in the north-south geometric direction, and high voltage lines have been shown to disrupt this body alignment suggesting that these animals can detect and respond to such an electromagnetic force (Burda et al., 2009).

Using topographic maps, we also recorded the amount of forest and cropland each deer encountered (based on radio locations) during an emigration. We then compared the per¬ cent forest and cropland present on the natal range with the same variables encountered along the travel corridor and with those within the newly settled home range using one-way ANOVA. Whether distance traveled between ranges affected final landscape selection was examined using distance traveled as a dependent variable in a stepwise regression analysis and using landscape characteristics found on the new range as independent variables (Table 2).

Home Range Characteristics

In an effort to determine the characteristics of the landscape in the 10 counties where our 25 females ultimately settled, we randomly selected 75 sections from these 10 counties using a ratio of 3 times the number of females followed from each study area (EC =14, NO = 5, WC = 6) (Anderson et al., 2005). The mean area of the new home range for our 25 females was 177.9 ± SE 41.9 ha (Table 3), and we used a circle of this size centered within each selected section on topographic maps to measure the amount of forests,

135

croplands, and grasslands present at the time of emigration. We then compared the percentage of forest, grasslands, and croplands found in these sections with the actual habitats selected by our marked females using one-way ANOVA.

Table 3. Mean values of landscape characteristics measured within the natal ranges, new home ranges, parturition ranges, and random sites of 25 female white-tailed deer in Illi¬ nois during 1980-1985 and 1990-1993. The natal and random sites were measured as the same new home range size.

						Percent of total		Forest patches only
Location	N	Home Range (ha)	Roads (km)	Houses (N)	Richness (all types)	Forest	Grassland	Crop	Size (ha)	Density (per 10 ha)	Nearest Neighbor (km)
Natal			^—i
	25	179.5	1.87	2.9	4.9	39.2	26.9	3 1 .3	52. 5a	0.12	0.08
New homi	2 (Ma	y 1 5 -Oet<	>ber 1 )			■ .	' ' mpi! ii§i <
	25	179.5	1.47	4.36	IT	35.7	19.7	43.4	20.7	0.29	0.13
Parturitioi	i (Ma	y 15-July	15)					HHHI			j 1 1 u gg in , mi .
	20	28.9	0 .30 1	().72:|	3.1	42.2	21.3	36.3	10.4	0.65a	0.09
Random sites									^ MflBM 1111 m m 1 Sill ' 1
	75	179.5	1.22	10.6	1.1	9.4	9.8	75.5		— No Data	—
aValue significantly different from other ranges/sites (see text)

We also used ANOVA tests to compare forest characteristics (Table 2) present in the natal ranges with the parturition and new home ranges selected by our marked females. Using one-way ANOVA, we compared similar characteristics of ranges selected by females (N = 15) that appeared to be stopped in their travels by an obstacle such as open agricultural land (e.g., a lack of forest), the presence of a large urban area, or a large river valley dominated by cultivated lands with those females (N = 10) where no obvious obstacle to further travel existed.

Costs of Emigration

Stepwise regression analysis using life span as the dependent variable was compared with 13 independent variables describing the human activities, general landscape characteris¬ tics found in the selected ranges, and possible social encounters (females present or not) (Table 2).

The possible effects of landscape selection on subsequent fawn survival was examined using one-way ANOVA to compare the landscape characteristics (total patch richness, forest patch nearest neighbor, forest patch density and average size) chosen by females that successfully raised 1 or more fawns with those that lost 1 or more fawns before fawn independence at 1 yr old.

RESULTS

Twenty-five radio marked females (4 adults, 7 yearlings, 14 fawns when marked) were tracked for an average of 20.9 mo. The usable radio fixes averaged 76.2 (range 7-286) per female (Table 1). The 14 dispersing females moved an average of 44.9 ± SE 6.4 km from their natal range, whereas the 1 1 migrators moved an average of 14.0 ± SE 7.2 km.

136

One female moved in February, 5 females moved in March, 5 in April, and 7 each in May and June. The February-April movements were made by females >20 mo old. The fawns (10-12 mo old) moved in May or June.

Nineteen of the 25 females were marked on the edge of the study areas where subordinate groups tended to reside (Nixon et al., 2010), and 18 of these 19 females were members of social groups considered subordinate. Only 5 females were offspring of a dominant matriarch or 1 of her daughters (social status was unknown for 2 females). Sixteen females were pregnant when they emigrated (6 pregnant for the first time), 5 were barren, and the reproductive status was unknown for the remaining 4 females. Twelve females left the natal range without kin support, whereas 13 traveled with a relative (2 with mother, 4 with a sibling, 7 with offspring of the previous year).

For the new home ranges selected by dispersing and migrating deer, there were no significant differences in the composition of home ranges selected by migrating (N = 1 1) versus dispersing (N = 14) females in the total patch richness, nearest neighbor distance, average forest patch density or size, forest type (upland vs bottomland), the percentage of landscape in forest, grassland, or cropland, or in the roads and houses present (r = 0.04- 1 .65, P = 0.1 1-0.98). Thus, we treated the 2 groups as a single sample.

Travel Characteristics

A comparison between a straight line drawn from the center of the natal range to the cen¬ ter of the new range indicated that travel deviated only 8.2% from a straight line. How¬ ever, the matched pair test indicated the actual path deviated by 2.33 ± SE 0.53 km and was significantly different from zero (P = 0.0003). For deer emigrating from the EC study area, with the general river corridor at 45°, the mean direction for the 14 females was 59° or eastward, a significant deviation from the direction of the river (P = 0.002).

Comparing the actual route with the simulated alternate routes (30°, 60°, or 90°), EC females crossed rivers significantly more often (F = 6.9, P = 0.009) and encountered significantly more forest patches (6.6 vs. 1 .8, F = 26.8, P < 0.0001), but did not differ in the number of roads (F = 0.74-1.52, P = 0.23-0.39) or power lines crossed (F = 0.04- 1.26, P = 0.39-0.85). Routes selected by females from the WC and NO areas, with no river corridor, did not differ from the simulated routes in the number of forests, rivers, roads, and high voltage power lines crossed during their travels.

A comparison between forest and cropland portions of the natal ranges, travel corridors, and new home ranges found that the travel corridor had more cropland (63% in travel corridor, 43% in new home range, 30% in natal range; F = 8.4, df = 2, 72, P = 0.0005), but all 3 locations were similar in the amount of forest cover encountered (35-42%), and all differed from random sites (P < 0.001) (Table 3).

The stepwise regression for distance traveled using 13 variables resulted in 3 significant variables (vegetative stage, movement type, and number of houses encountered) account¬ ing for nearly all the variance (R: = 0.83). However, these results were biased by the longer distances moved by dispersing females and the disparity in initiation of movement between fawns (May-June) and older deer (February-April). The farther deer dispersed,

137

the more houses were present in the selected new home range, perhaps an indication that these females were becoming less discriminating as parturition approached.

Home Range Characteristics

Females selected a new home range with significantly more forest (35.7% vs. 9.0%), more permanent grassland (19.7% vs. 2.2%), and less cropland (43.5% vs. 75.5%) than was indicated as available in our random sections from 10 counties where the 25 females selected a new home range (F = 4.7-34.2, P = 0.03-< 0.001) (Table 3). Females in EC Illinois settled on areas with more total patch richness, forest patch density, and forest patch size compared to random sites (P = 0.007). Northern Illinois females also selected sites featuring more forest patches of larger size (P = 0.01) compared to random sites. Home range size was positively associated with the percent of cropland present in new home range (F = 10.6, P = 0.003) and negatively associated with the percent of forest cover present (F = 7.3, P = 0.01). This result was biased by the large ranges occupied by the 3 females that settled on landscapes entirely devoid of forest cover. However, final home range size was not influenced by patch richness, forest patch number and size, for¬ est type and age, or the distance between forest patches (P > 0.05) (Table 3).

On new home ranges, the presence or absence of conspecific females (17 present, 8 absent) did not differ as to patch richness, nearest neighbor distance, size or density of forest patches, or with random sites (Table 3). In a comparison of natal, new home, and parturition ranges, patch richness was significantly higher in the new home range (F = 7.6, df = 2, 65, P = 0.001) (Table 3). There were more forest patches present in the parturition ranges than in the other 2 ranges (F = 15.7, df = 2, 64, P < 0.0001), but the natal ranges contained larger patches (F = 15.2, df = 2, 62, P < 0.001). Parturition sites featured fewer houses (P = 0.04) and roads (P < 0.001) than the natal or new home ranges.

Five females stopped emigrating because their way was blocked by an obvious obstacle, 3 by an urban area, 1 by the expansive Interstate-72 highway which she never crossed, and 1 female by the 1.6 km wide cultivated Illinois River valley (Table 3). An additional 7 females traveled within a watershed that progressively lost forest cover. These females stopped at the site of the last reasonably abundant forest cover after first probing ahead and then returning along the more open watershed. Six additional females stopped in the first woodlot encountered after leaving the natal range. There was no obvious reason for stopping for 7 females (5 pregnant, 1 barren, 1 reproductive status unknown) other than, in some cases, impending parturition for 5 of 7 females. We compared the landscape composition of home ranges of females stopped by an obvious obstacle (N = 5) with those that were not stopped (N = 20) to see if females that stopped had home range characteristics different from those with a more open choice. We found no significant difference in the amount of forest, grasslands, and croplands present between the 2 groups (P > 0.05).

Costs of Emigration

We used life span and fawn survival to assess the costs of locating away from the natal range and the loss of kin support. Stepwise regression using life span as the dependent variable and 13 landscape and human intrusion variables occurring in each new home range, found only the type of movement (disperse or migrate) significant (R = 0.50). We

138

did not find any significant effect of composition in the new home range on whether or not our females lost 1 or more fawns before weaning (P > 0.20). Of 6 pregnancies (by 5 females) where fawns were lost, 4 occurred by the end of August.

DISCUSSION

Emigrating females in WC Illinois did not select habitats significantly different in composition from random sites. Apparently when forest cover is abundant (in this case about 20% of the landscape), dispersing female's selection of a parturition site depends on factors other than habitat composition such as obstacles encountered, impending birth, or avoiding human intrusions. However, the more forested landscape in WC Illinois did not reduce the distance females traveled before settling on a new home range (Table 1).

In areas of Illinois where forest covers <5% of the landscape, emigrating females must search for forest cover sufficient to protect fawns from disturbance and predation. Marked females from EC and NO Illinois selected a landscape with more forest and permanent grassland present than was available in the general landscape. Three NO females (#159, #155, #196) each selected a range with no permanent forest cover except fence lines and narrow hedgerows. Two of these were from subordinate social groups, and both of these deer were pregnant. The remaining female dispersed from a dominant social group, her immediate reproductive status unknown. Selections of forest-free areas indicate some females in Illinois appear to use agricultural lands in a similar fashion as roe deer do in Europe, including settling in areas entirely devoid of forests (Harris and Woollard, 1990).

Home range habitat characteristics selected by migrating and dispersing females differed only in the distance between forest patches, with migrating females selecting the more scattered forest cover. These migrating females did not have to winter on these ranges and could accommodate a summer-fall range with less forest cover. Migratory females live longer than dispersing females, a result of their vacating hunted summer ranges for the more protected natal range (Nixon et al., 2008). We do not know how or when deer determine the suitability of a site for year-round occupancy, but selection of a cover-defi¬ cient site necessitates movement away in late fall. We suspect deer do not initially deter¬ mine this suitability but depend on circumstances to dictate a movement response as the seasons change. The return to the natal range is the easier choice because females are familiar with the route back to the natal range.

Both types of movement behavior used nearly a straight line during emigration, readily crossing rivers, roads, and high voltage power line rights-of-way. Travel corridors trav¬ ersed significantly more cropland than was present in the selected new home range, an expected result given Illinois’ agriculturally dominated landscape. Travel distance was influenced by plant phenology (farther distance after leaf out), but age was the primary cause, with fawns moving later and farther than older deer.

There is almost no literature on how emigrating deer orient themselves during a dispersal movement (Andreassen et al., 2002). Long et al. (2010) found that male immigrants were influenced in their selection of a travel route by the presence of mountain ridges on 1 study area in Pennsylvania but not from a second area. These Pennsylvania deer also

139

tended to avoid 4-lane highways, a feature of emigration detected for only 1 of our females (#262). The presence of roads can alter behaviors by affecting home range size, escape responses, and physiology for deer exposed to them (Trombulak and Frissell, 2000), but Illinois deer are exposed to many road crossings almost from birth. The natal ranges for our study females were surrounded by roads with at least moderate traffic.

A forested travel corridor existed within the EC study area but not on the WC and NO study areas, and 10 of 14 EC females used the river corridor to exit their natal range. Four of these females then left the river corridor and traversed agricultural fields to the site of new home range selection. Three of these females selected ranges without other females present, perhaps because of resident female harassment during travel or that 3 of these females were considered subordinate in the social order on their natal range. One female (#507) was born into a dominant group, but also selected a female-free location for a parturition site.

Most of our study females moved rapidly and in nearly a straight line between the natal and final home range, considered the simplest and most efficient way to complete an emigration movement (Zollner and Lima, 1999; Wiems, 2001). Long et al. (2010) also observed that emigrating male white-tails moved quickly without much wandering and, because of this behavior, felt that males were not searching for specific habitats. Kilgo et al. (1996) found that dispersing deer also traveled rapidly in nearly a straight line while moving through unfamiliar landscapes in Florida. There appeared to be no avoidance of rivers, major roads, or high voltage power lines during female travels between ranges in Illinois. In fact, 10 of 25 females settled on ranges with a high voltage power line present and were seen bedded and feeding within the power line rights-of-way. Except during flooding episodes, rivers encountered were generally <50 m wide and presented no obsta¬ cle to continued movement.

Parturition ranges for all 25 females contained fewer houses and roads and more forest patches than either the natal ranges or the larger new home ranges. Storm et al. (2007) reported that breeding females avoided occupied houses during the parturition season in southern Illinois. More forest cover has been associated with increased fawn survival in Illinois (Rohn et al ., 2007) and a somewhat higher ratio of resident females to migrating females in the northern plains (Grovenburg et al., 2011). Coyotes are less efficient hunt¬ ers in forest cover, so more forest cover should improve fawn survival (Richer et al ., 2002; Rohn et al., 2007). Rohn et al. (2007) found that more edge and larger forest patches improved fawn survival by reducing predator risk in southern Illinois. They also noted that smaller forest patches surrounding the larger forest blocks reduced home range size and provided more feeding opportunities (Rohn et al., 2007). Our marked females also selected ranges for increased patch density and fewer human activities in the parturi¬ tion range. Fawn survival appeared to be independent of landscape composition, similar to observations in Pennsylvania where patch diversity, amount of forest edge, and road density had no significant affect on fawn survival (Vreeland et al., 2004).

Life span of females was affected by the amount of forest cover, with larger forests more likely to offer increased hunting opportunities and a lower life expectancy. This is the reverse situation found for deer in South Dakota where more forest cover improved life span (Klaver et al., 2008). In Illinois, even the larger forest patches were generally <100

140

ha in size (Iverson et al., 1989). Harden et al. (2005) found that the larger blocks of forest cover attracted more deer hunters in southern Illinois.

The new home range selected by our marked females tended to be elongate in shape, considered an optimum shape for exploiting a patchy environment (Ford, 1983). Home range size was affected by the amount of cropland encountered with females encompass¬ ing a much larger home range as forest cover declined, similar to that observed in SW Minnesota (Walter et al., 2009). Home range size for the 3 females with no forest cover present ranged up to 1,000 ha with daily movements often 2-3 km, similar to home ranges and movements described for deer in South Dakota (Sparrowe and Springer, 1970; Grovenburg et al., 2009). Two of the 3 females were migrants, returning to their natal range each winter after corn harvest. The third female sheltered in unpicked corn during winter.

A first pregnancy for 6 females that delayed emigration until late May-early June meant that these females did not have a lot of time before parturition to find a new range. As noted by Orians and Wittenberger (1991), the later emigration is delayed before some important event (parturition, winter weather, etc.), the poorer the options available. Estro¬ gen levels continue to increase from 6 wk prepartum to parturition, and the estro¬ gen/progesterone ratio rises steeply during the final weeks of pregnancy (Plotka et al., 1977) which may stimulate pregnant females to disperse quickly close to parturition. Five of these females were dispersers, and 1 was a migrant. Even though forest cover was quite limited for 3 of these females, all 6 reared fawns to weaning age, and 2 of these females survived >2 yr on their new range.

Some of the marked fawns and yearlings departed the natal range without kin support (10 moved alone, 2 with unknown deer), relying on instinct and perhaps the knowledge gained by familiarity with the characteristics of the natal range (forest age and composi¬ tion) to guide their selection process. Forests surrounding all sites were very similar to the natal ranges for these females in age and flooding frequencies (Nixon et al., 1991). Deer obviously use olfactory cues, sounds emanating from the environment, and their vision to settle on or avoid areas as they travel (Andreassen et al., 2002). Six females had favorable winds blowing from the site of final settlement during at least 2 days while emigrating.

Six of our 25 females were known to have stopped for 1-3 days along the journey, per¬ haps accessing the qualities of the site for permanent occupancy (Opdam, 1990; Stamps, 2001). Emigration then continued perhaps because of resident deer aggression (Ozoga et al., 1982), insufficient cover before crops are high enough for concealment before a June movement (Nixon et al., 2007), insufficient forage (unlikely in an agriculture-dominated landscape), excessive human disturbance (Etter, 2001), or, for 5 of the 6 females, impending parturition.

Dispersal theory states that dispersing individuals should disperse to the first vacant terri¬ tory and stop because longer movements incur more risks (Murray, 1967). However, in areas where forests cover is <5% of the landscape and human disturbances are frequent, dispersing females are forced to travel long distances before settling (Sparrowe and Springer, 1970; Zagata and Haugen, 1973; Kernohan et al., 1994; Nixon et al., 2007).

141

Migrating females move shorter distances and can afford to be less selective as to cover because they return to their natal range in winter. Unlike males, females, because of fawn rearing requirements, must be more selective in order to protect fawns when selecting a new range. Life threatening events such as hunting are not apparent when females select new ranges in late spring-early summer, but even so, life history attributes like dispersal or migratory behaviors that evolved as protection from predation, variable climates, or habitat alterations from the Pleistocene to the present day are still effective options in today’s environment in Illinois (Danchin et al ., 2001 ; Stamps, 2001).

ACKNOWLEDGEMENTS

This study would not have been possible without the able assistance of J. M. Nelson, M. Challand, S. Rueff, M. E. Billman, R. Sawtelle, M. Nelson, W. Iko, J. Cruise, R. Conover, B. Martin, S. White, D. Tazik, J. Dewalt, D. Dillow, G. Potts, R. Seimers, J. Brown, M. Schmierbach, J. Legare, C. Nixon, J. Nixon, J. McNamara, J. Bothwell, R. Harrison, R. Koerkenmeir, and J. Ver Steeg who volunteered numerous times to capture deer and hunt for new born fawns. J. Seets, Illinois Natural History Survey, assisted in many ways. D. Bowman, J. Assell, and J. Sandine, site superintendents of Robert Aller- ton Park, Siloam Springs State Park, and Shabbona Lake State Park, respectively, pro¬ vided study areas and logistical support. This paper is a contribution (in part) of Federal Aid in Wildlife Restoration Project W-87-R, The Illinois Department of Natural Resources, U.S. Fish and Wildlife Service, and the Illinois Natural History Survey cooperating.

LITERATURE CITED

Anderson, D. P., M. G. Turner, J. D. Forester, J. Zhu, M. S. Boyce, H. Beyer and L. Stowell. 2005. Scale-dependent summer resource selection by reintroduced elk in Wisconsin, USA. J. Wildl. Manage., 69:296-310.

Andreassen, H. P., N. C. Stenseth and R. A. Ims. 2002. Dispersal behavior and population dynam¬ ics of vertebrates, p. 237-256. In: J. M. Bullock, R. E. Kenwood and R. S. Hails (eds.). Dispersal Ecology. Blackwell Publ., Malden, Massachusetts.

Brinkman, T. J., C. S. Deperno, J. A. Jenks, B. S. Haroldson and R. G. Osborn. 2005. Movement of female white-tailed deer: effects of climate and intensive row-crop agriculture. J. Wildl. Man¬ age., 69:1099-\UI.

Burda, H., S. Begall, J. Cerveny, J. Neef and P. Nemec. 2009. Extremely low-frequency electromagnetic fields disrupt magnetic alignment of ruminants. PNAS, 106:5708-5713.

Clutton-Brock, T. H., F. E. Guinness and S. D. Albon. 1982. Red deer: behavior and ecology of two sexes. University of Chicago Press, Chicago, Illinois.

Danchin, E. D., D. Heg and B. Doligez. 2001 . Public information and breeding habitat selection, p. 243-258. In: J. Clobert, E. Danchin, A. Dhondt and J. Nichols, (eds.) Dispersal. Oxford Univer¬ sity Press, New York.

Dusek, G. L., R. J. Mackie, J. D. Herriges, Jr. and B. B. Compton. 1989. Population ecology of white-tailed deer along the lower Yellowstone River. Wildl. Monog., 104. The Wildlife Society, Washington, D.C.

Etter, D. R. 2001. Ecology and management of overabundant white-tailed deer from suburban Chi¬ cago, Illinois. Ph.D. Dissertation, University of Illinois, Urbana, Illinois.

Etter, D. R., C. M. Nixon, J. B. Sullivan and J. A. Thomas. 1995. Emigration and survival of orphaned female deer in Illinois. Can. J. Zool., 73:440-445.

Ford, R. G. 1983. Home range in a patchy environment: optimal foraging predictions Amer. Zool., 23:315-326.

142

Gladfelter, H. L. 1978. Movement and home range of deer as determined by radio telemetry. Iowa Wildl. Res. Bull, 23.

Godvik, I. M. R., L. E. Loe, J. O. Vik, V. Veiberg, R. Langvator and A. Mysterud. 2009. Temporal scales, trade-offs, and functional responses in red deer habitat selection. Ecology, 90:699-710.

Grovenburg, T. W., J. A. Jenks, R. W. Klaver, C. C. Swanson, C. N. Jacgues and D. Todey. 2009. Seasonal movements and home ranges of white-tailed deer in north-central South Dakota. Can. J.Zool, 87:876-886.

Grovenburg, T. W., J. A. Jenks, R. W. Klaver, C. S. DePerno, T. J. Brinkman, C. C. Swanson and J. A. Jenks. 2011. Influence of landscape characteristics on migration strategies of white-tailed deer. J. Mammal, 92:534-543.

Grovenburg, T. W., C. N. Jacques, C. C. Swanson, R. W. Klaver and J. A. Jenks. 2010. Use of late season standing corn by female white-tailed deer in the northern great plains during a mild win¬ ter. Prairie Nat., 42:8-18.

Harden, C. D., A. Woolf and J. Roseberry. 2005. Influence of exurban development on hunting opportunity, hunter distribution, and harvest efficiency of white-tailed deer. Wildl. Soc. Bull., 33:233-242.

Harris, S. and T. Woollard. 1990. The dispersal of mammals in agricultural habitats in Britain, p. 159-188. In: R. Bunce and D. Howard (eds). Species dispersal in agricultural habitats. Belhaven Press, New York.

Hawkins, R. E., L. D. Martoglio and G. G. Montgomery. 1968. Cannon netting deer. J. Wildl. Man¬ age., 32:191-195.

Illinois Commerce Commission. 1971. Electric utilities in Illinois, 34,500 or higher voltage. Ill. Commerce Comm., Springfield.

Illinois Commerce Commission. 1997. Electric utilities in Illinois, 34,500 or higher voltage. Ill. Commerce Comm., Springfield.

Iverson, L. R., R. L. Oliver, D. P. Tucker, P. G. Risser, C. D. Burnett and R. G. Rayburn. 1989. The forest resources of Illinois: an atlas and analysis of spatial and temporal trends. III. Nat. Hist. Surv. Spec. Publ., 11:181.

Kilgo, J. C., R. F. Labisky and D. E. Fritzen. 1996. Directional long-distance movements by white¬ tailed deer ( Odocoileus virginianus ) in Florida. Wildlife Biology, 3:289-292.

Kernohan, B. J., J. A. Jenks and D. E. Naugle. 1994. Movement patterns of white-tailed deer at Sand Lake National Wildlife Refuge, South Dakota. Prairie Nat, 26:293-300.

Klaver, R. W., J. A. Jenks, C. S. Deperno and S.L. Griffin. 2008. Associating seasonal range characteristics with survival of female deer. J. Wildl. Manage., 72:343-353.

Lesage, L., M. Crete, J. Huot and J. P. Ouellet. 2002. Use of forest maps versus field surveys to measure summer habitat selection and sexual segregation in northern white-tailed deer. Can. J. Zool., 80:717-726.

Long, E. S., D. R. Diefenbach, B. D. Wallingford and C. S . Roseberry. 2010. Influence of roads, rivers, and mountains on natal dispersal of white-tailed deer. J. Wildl. Manage., 74:1242-1249.

Morrison, M. L., B. G. Plarcot and R. W. Mannan. 2006. Wildlife-habitat relationships: concepts and applications. 3rd edition. Island Press, Washington, D.C.

Murray, B. G., Jr. 1967. Dispersal in vertebrates. Ecology, 48:975-978.

Nams, V. O. 1990. Locate II user’s guide. Pacer, Truro, Nova Scotia, Canada.

Nelson, M. E. and L. D. Mech. 1992. Dispersal in female white-tailed deer. J. Mammal., 73:89 1— 894.

Nixon, C. M., L. P. Hansen, P. A. Brewer and J. E. Chelsvig. 1991. Ecology of white-tailed deer in an intensively farmed region of Illinois. Wildl. Monog. 118. The Wildlife Society, Washington, D.C.

Nixon, C. M., L. P. Hansen, P. A. Brewer and J. E. Chelsvig. 1992. Stability of white-tailed doe parturition ranges on a refuge in east-central Illinois. Can. J. Zool., 70:968-973.

Nixon, C. M., P. C. Mankin, D. R. Etter, L. P. Hansen, P. A. Brewer, J. E. Chelsvig, T. L. Esker and J. B. Sullivan. 2007. White-tailed deer dispersal behavior in an agricultural environment. Amer. Midi. Nat., 157:212-220.

Nixon, C. M., P. C. Mankin, D. R. Etter, L. P. Hansen, P. A. Brewer, J. E. Chelsvig, T. L. Esker and J. B. Sullivan.. 2008. Migration behavior among female white-tailed deer in central and northern Illinois. Amer. Midi Nat., 160:178-190.

143

Nixon, C. M., P. C. Mankin, D. R. Etter, L. P. Hansen, P. A. Brewer, J. E. Chelsvig, T. L. Esker and J. B. Sullivan.. 2010. Characteristics of dominant and subordinate led social groups of white-tailed deer in Illinois. Amer. Midi. Nat., 163:388-399.

Opdam, P. 1990. Dispersal in fragmented populations: the key to survival, p. 3-17. In: R. G. H. Bunce and D. C. Howard, (eds.) Species dispersal in agricultural habitats. Belhaven Press, New York.

Orians, G. H. and J. F. Wittenberger. 1991. Spatial and temporal scales in habitat selection. Amer. Nat., 137:529-549.

Ozoga, J. J., L. J. Verme and C. S. Bienz. 1982. Parturition behavior and territoriality in white¬ tailed deer: impact on neonatal mortality. J . Wildl. Manage., 46: 1-1 1 .

Plotka, E. D., U. S. Seal, L. J. Verme and J. J. Ozoga. 1977. Reproductive steroids in the white-tailed deer ( Odocoileus virginianus borealis). II. Progesterone and estrogen levels in peripheral plasma during pregnancy. Biol. Reprod., 17:78-83.

Richer, M. C., M. Crete, L. P. Rivest and J. Huot. 2002. The low performance of forest versus rural coyotes in northern North America: inequality between presence and availability of prey. Ecosci- ence, 9:44-54.

Rohn, J. H., C. K. Neilsen and A. Woolf. 2007. Survival of white-tailed deer fawns in southern Illinois. J . Wildl. Manage., 71:851-860.

Severinghaus, C. W. 1949. Tooth development and wear as criteria of age in white-tailed deer. J. Wildl. Manage., 13:195-216.

Sparrowe, R. D. and P. F. Springer. 1970. Seasonal activity patterns of white-tailed deer in eastern South Dakota. J. Wildl. Manage., 34:420-431 .

Stamps, J. A. 2001. Habitat selection by dispersers: integrating proximate and ultimate approaches, p. 230-242. In: J. Clobert, E. Dauchin, A. A. Dhondt and J.D. Nichols (eds.). Dispersal. Oxford University Press, New York.

Storm, D. J., C. K. Nielsen, E. M. Schulier and A. Woolf. 2007. Space use and survival of white¬ tailed deer in an exurban landscape. J. Wildl. Manage., 71:1 170-1 176.

Tierson, W. C., G. F. Mattfeld, R. W. Sage, Jr. and D. F. Behrend. 1985. Seasonal movements and home ranges of white-tailed deer in the Adirondacks. J. Wildl. Manage., 49:760-769.

Trombulak, S. C. and C. A. Frissell. 2000. Review of ecological effects of roads on terrestrial and aquatic communities. Conserv. Biol., 14:18-30.

VerCauteren, K. C. and S. E. Hygnstrom. 1994. A review of white-tailed deer movements in the Great Plains relative to environmental conditions. Great Plains Research, 4:1 17-132.

Vreeland, J. K., D. R. Diefenbach and B. D. Wallingford. 2004. Survival rates, mortality causes, and habitats of Pennsylvania white-tailed deer fawns. Wildl. Soc. Bull., 32:542-553.

Walter, W. D., K. C. VerCauteren, J. M. Gilsdorf and S. E. Hygnstrom. 2009. Crop, vegetation and biofuels: response of white-tailed deer to changing management priorities. J. Wildl. Manage., 73:339-344.

Wiems, J. A. 2001 . The landscape context of dispersal, p. 96-109. In: J. Clobert, E. Danchin, A. A. Dhondt and J. Nichols (eds.). Oxford University Press, New York.

Wood, A. K., R. E. Short, A. Darling, G. L. Dusek, R. G. Sasser and C. A. Ruder. 1986. Serum assays for detecting pregnancy in mule and white-tailed deer. J . Wildl. Manage., 50:684-687.

Zagata, M. D. and A. O. Haugen. 1973. Winter movement and home range of white-tailed deer at Pilot Knob State Park, Iowa. Proc. Iowa Acad. Sci., 79:74-78.

Zar, J. H. 1999. Biostatistical analysis. Prentice Hall, New Jersey.

Zollner, P. A. and S. L. lima. 1999. Search strategies for landscape level interpatch movements. Ecology, 80:1019-1030.

144

Transactions of the Illinois State Academy of Science (2012) Volume 105,#3&4 pp. 145-152

received 5/19/12 accepted 12/3/12

Illinois Status Survey of the Redside Dace Clinostomus elongatus: The Newest Addition to the State's Native Fish Fauna

Jeremy S. Tiemann* and Mark H. Sabaj Perez1 Illinois Natural History Survey, Prairie Research Institute University of Illinois, 1816 South Oak Street, Champaign, IL 61820 ’Present Address: The Academy of Natural Sciences of Drexel University 1900 Benjamin Franklin Parkway, Philadelphia, PA 19103-1 195 Correspondence: jtiemann@illinois.edu

ABSTRACT

The redside dace Clinostomus elongatus is a small, laterally compressed cyprinid com¬ monly found in small streams with moderate to high gradients, clear and cool water, and substrates of clean gravel, sand, or bedrock. Fish surveys in Winnebago County, Illinois, and Rock County, Wisconsin, conducted from 1997-2000 and 2010-2011 discovered the dace at a single site in Illinois in East Fork Raccoon Creek (Pecatonica River - Rock River Drainage). This is the first documented record of this species in Illinois and raises the known total of state native fishes to 192. Based on these surveys, C. elongatus is expected to periodically occur in the Illinois portion of the Raccoon Creek basin, and therefore should be considered a peripheral species in Illinois.

INTRODUCTION

The redside dace Clinostomus elongatus is a small, laterally compressed minnow (Cyprinidae) with a large oblique mouth and long pointed snout. It commonly inhabits small streams with moderate to high gradients, clear and cool water, and substrates of clean gravel, sand, or bedrock (Trautman, 1981; Becker, 1983). The dace is drawn to particular habitat features, and generally orients toward stream positions with slower cur¬ rent velocities, greater depths, and closer to woody debris (Novinger and Coon, 2000; Zimmerman, 2009). Clinostomus elongatus is distributed across once glaciated regions of the northeastern United States and southern Ontario in watersheds draining into the Mississippi River, Ohio River, Great Lakes, and Atlantic Ocean (COSEWIC, 2007). The western-most populations occur in a few isolated, disjunct populations throughout the upper Mississippi River basin, including the Raccoon Creek basin (Pecatonica River - Rock River Drainage) in Rock County, Wisconsin (Gilbert, 1980; Becker, 1983). How¬ ever, the species had never been documented in Illinois. We performed fish surveys in the Raccoon Creek basin to determine the presence of C. elongatus in Illinois.

146

METHODS

Two assessment surveys were preformed in the Raccoon Creek basin (Pecatonica River - Rock River Drainage). The first assessment occurred from November 1997 to April 2000 and was conducted by MHSP (Sabaj, 2000), whereas the second was conducted by JST and occurred from June 2010 to October 2011. During the first assessment, fishes were collected at 10 sites in the basin with a 2.43 m minnow seine and a Smith-Root backpack electrofisher (model 12-B) powered by a 24 volt, 12 Ah battery for up to 4 hours per site; each site was visited 1-5 times during the Spring-Summer and/or Fall-Winter (Sabaj, 2000). During the second assessment, fishes were collected at 20 sites (Table 1) for up to 2 hours per site using a barge electro-shocker set at 200 volts, a Smith-Root Model 12 DC backpack electrofisher, or a 3.05 m minnow seine; each site was visited four times. During each site-visit, all fishes collected were identified to species in the field. A subsample of fishes representing the total diversity collected at each site was vouchered and deposited in the Illinois Natural History Survey Fish Collection, Champaign (INHS). Fishes collected but not vouchered were returned without harm to their native habitat. Nomenclature follows Page and Burr (2011). In addition, the Wisconsin Department of Natural Resources’ (WDNR) Fish Mapper was accessed (http://infotrek.er.usgs.gov /wdnrfish/map/index), and fishes housed at the following museums were searched for voucher specimens of C. elongatus collected from Illinois and Wisconsin: Academy of Natural Sciences, Philadelphia (ANSP), Field Museum of Natural History, Chicago (FMNH), INHS, Milwaukee Public Museum (MPM), University of Kansas Museum of Natural History, Lawrence (KU), Southern Illinois University Fish Collection, Carbon- dale (SIUC), University of Michigan Museum of Zoology, Ann Arbor (UMMZ), and United States National Museum - Smithsonian Institution, Washington D.C. (USNM). Museum names and codes follow Sabaj Perez (2012).

RESULTS

The two assessments of the Raccoon Creek basin yielded 55 native species of fishes, including C. elongatus (Table 1). However, only once was C. elongatus collected in Illi¬ nois. Eight juveniles were taken on 31 May 1998 in East Fork Raccoon Creek, 4.3 km NW Rockton, Winnebago County, 42.48508°N, 89.13744°W (Figure 1 - Site 17). Seven specimens were vouchered (INHS 46430) and one was returned live to the stream at the place of capture. All specimens were collected in a relatively deep (ca. 3-3.5 ft.) and par¬ tially shaded pool (side pocket of the main channel) with a thick silt and mud substrate. In the previous year, a beaver dam stretched across the channel just below the mouth of the pocket, inundating a short stretch of the creek. The beaver dam had been removed by the landowner prior to the 1998 visit; however, the pool retained water and provided ref¬ uge for the redside dace and many other juvenile minnows including bigmouth shiners Notropis dorsalis , southern redbelly dace Chrosomus erythrogaster , fathead minnows Pimephales promelas, creek chubs Semotilus atromaculatus and brassy minnows Hybog- nathus hankinsoni. The same pool was sampled on eight additional occasions, once before collecting the redside dace and seven times after, but only the one visit yielded any dace.

147

Table 1. Sampling sites for the Clinostomus elongatus survey. Streams include Raccoon Creek (RC), unnamed tributaries to Raccoon Creek (Trib RC), East Fork Raccoon Creek (EFRC), and unnamed tributary to East Fork Raccoon Creek (Trib EFRC). Latitude and longitude are in decimal degrees. Reference Point is approximate location of site on gazetteer. Point on map (PoM) refers to site number on Figure 1 . Asterisks (*) are the sites sampled by Sabaj (2000). Last observed (Last obs.) is when Clinostomus elongatus was last observed at the site and includes the 1997-2000 and 2010-201 1 surveys as well as those preformed by Fago (1982).

Stream	State: County	Latitude	Longitude	Reference Point	PoM	Last obs.
RC	WI: Rock	42.5751	-89.2079	S Luther Valley Rd	1	Fago (1982)
RC	WI: Rock	42.5416	-89.2072	W Beloit Newark Rd	2	2011
RC	WI: Rock	42.5265	-89.1955	WI Rte 81	3
RC	WI: Rock	42.5092	-89.1696	Mill Pond Rd	4	Fago (1982)
RC	WI: Rock	42.5038	-89.1609	W St Lawrence Rd	5
trib RC	WI: Rock	42.5234	-89.1781	WI Rte 81	6
trib RC	WI: Rock	42.5278	-89.1692	Co Hwy H	7
EFRC	WI: Rock	42.5628	-89.1446	W Gravedale Rd	8	Fago (1982)
EFRC	WI: Rock	42.5408	-89.1326	W Beloit Newark Rd	9	2011
EFRC	WI: Rock	42.5261	-89.1256	W Spring Creek Rd	10*	2011
EFRC	WI: Rock	42.5041	-89.1173	W St Lawrence Rd	11	2011
EFRC	WI: Rock	42.4974	-89.1202	State line	12*	2011
trib EFRC	WI: Rock	42.5255	-89.0949	W Spring Creek Rd	13*
RC	IL: Winn.	42.4916	-89.1482	Power line crossing	14*
RC	IL: Winn.	42.4760	-89.1377	Yale Bridge Rd	15*
RC	IL: Winn.	42.4557	-89.1270	W Rockton Rd	16*
EFRC	IL: Winn.	42.4850	-89.1374	Confluence with RC	17*	1998
Trib RC	IL: Winn.	42.4939	-89.1708	Pomeroy Rd	18*
Trib RC	IL: Winn.	42.4768	-89.1266	Yale Bridge Rd	19*
Trib RC	IL: Winn.	42.4676	-89.1381	Clover Rd	20*

The dace was collected throughout the Wisconsin side of the Raccoon Creek basin including <0.5 km from the state line during the second survey (Table 1). Three adults were collected on 1 August 2011 in East Fork Raccoon Creek, 7.3 km W Beloit, Rock County, 42.49969°N, 89.11987°W (Figure 1 - Site 12). One specimen was vouchered (INHS 104899) and two were returned live to the stream at the place of capture. These three individuals were collected in a clear pool containing gravel/cobble substrates. This site was ~1 km upstream of where the Illinois specimens were collected in 1998. Of the museums examined, only INHS and MPM had redside dace specimens from the Raccoon Creek basin. On 8 June 1998, 24 adult specimens were collected in the East Fork Rac¬ coon Creek, 8.0 km W Beloit, in Rock County, Wisconsin, 42.52617°N, 89.12562°W (INHS 46977). MPM had specimens collected by Fago (1982), who reported the dace from another locality in the upper East Fork and 3 localities in upper Raccoon Creek, Rock County, Wisconsin (Table 1; Figure 1). According to the WDNR’s Fish Mapper, the only stream flowing into Illinois that contained C. elongatus was Raccoon Creek.

148

Figure 1. Map of the study area. Solid stars indicate sites where Clinostomus elongatus was collected during the 1997-2000 and 2010-201 1 surveys, open stars show where Fago (1982) reported the fish, and open circles designate those sites where we failed to collect the dace during our surveys.

DISCUSSION

This study is the first to document C. elongatus in Illinois, and increases the total number of fish species reported as native to Illinois waters to 192 (Burr and Page, 2009). Resident populations of C. elongatus occur in the Wisconsin headwaters of East Fork Raccoon Creek and Raccoon Creek; however, the dace is evidently a peripheral species in Illinois. The substrate changed from predominantly gravel and cobble over mud and sand on the Wisconsin side of the basin to largely sand with scattered patches of gravel and small cobble in Illinois. Although its occurrence in the state has been confirmed, it remains undetermined whether this species reproduces in Illinois and maintains a resident popula¬ tion or is just an occasional transient that is washed downstream.

As reported by Koster (1939), C. elongatus was commonly collected with blacknose dace Rhinichthys atratulus , creek chubs, white suckers Catostomus commersoni, and Johnny

149

darters Etheostoma nigrum. Redside dace typically reproduce over the pebble-nests of other minnows in small headwater streams dominated by gravel substrates (Koster, 1939). Two pebble nest-building species (hornyhead chub Nocomis biguttatus and creek chub) were found throughout the Raccoon River basin in Illinois. However, habitat suita¬ ble for pebble-nests (e.g., clean gravel substrates) was rather uncommon in the Illinois portion of this system, which is dominated by sand. Two small tributaries to Raccoon Creek (see sites 18 and 20, Figure 1) offered the best habitats for pebble-nests and are thereby the best candidates for supporting viable Illinois populations of C. elongatus. However, these streams were often dry and collections in these tributaries yielded no specimens.

Several other state native fishes have been discovered since Smith's (1979) comprehen¬ sive summary of the state's ichthyofauna. The bleeding shiner Luxilus zonatus , taillight shiner Notropis maculatus, and fringed darter Etheostoma crossopterum were not reported in Smith (1979) but have been collected in Illinois in the last 30 years (Burr et al., 1988; Poly and Wilson, 1998; Hiland and Poly, 2000). The cypress minnow Elybog- nathus hayi , bigeye chub Hybopsis amblops, and crystal darter Cystallaria asprella were considered extirpated by Smith (1979), but since have been sporadically collected in Illi¬ nois (Warren and Burr, 1989; Burr et al., 1996; Tiemann et al., 2004; Stewart et al., 2005; Steuck et al., 2010). Although the northern studfish Eundulus catenatus is listed as part of the state’s ichthyofauna, Smith (1979) stated that there was no evidence of a population in Illinois and the species was a “straggler from a Missouri tributary.” A single specimen was collected in July 2007 in the West Fork Richland Creek (Kaskaskia River Drainage), St. Clair County (Randy Sauer, Illinois Department of Natural Resources, personal communication). These recent additions to the biodiversity of Illinois provide a refresh¬ ing, albeit fleeting departure from more common statewide trends such as species extirpa¬ tions (e.g., Burr and Warren, 1986) and invasions by non-indigenous fishes (e.g., Laird and Page, 1996; Chick et al., 2003; Irons et al., 2006).

The Pecatonica River drainage, which includes the Raccoon Creek basin, has seen an increase in native fish species richness during the last 100 years (Retzer, 2005). In an evaluation of Illinois streams based on aquatic biodiversity, Page et al. (1992) listed Rac¬ coon Creek as a Biologically Significant Stream so it is no surprise that the basin sup¬ ports a diverse fish fauna, including the Illinois state-threatened starhead topminnow Fundulus dispar and Iowa darter Etheostoma exile. Though not formally sampled, we also encountered a diverse mussel fauna with 10 live species, including the Illinois state- threatened slippershell Alasmidonta viridis. However, the Pecatonica River basin, includ¬ ing Raccoon Creek, has been threatened by siltation and agricultural pollution (Smith, 1971; Page et al., 1992). Additionally, Raccoon Creek is altered by a headwater impoundment (Mill Pond) near our Site 4. In addition to altering habitat and blocking fish dispersal, dam effects include stocking of sportfish (Taylor et al., 2001; Tiemann et al., 2007). We collected several top predators near Site 4, including largemouth bass Microp- terus salmoides and northern pike Esox lucius, that were not collected elsewhere in the basin.

Multiple authors have noted a decrease in the overall range and abundance of the redside dace, primarily as a result of activities that increase turbidity, silt deposition, and mean water temperature in small streams (either as a result of dams, climate change, or removal

150

of riparian areas), or introduction of top predators (Harlan and Speaker, 1956; Trautman, 1981; Lyons et al., 2000; COSEWIC, 2007). Because of these threats and its limited distribution, the American Fisheries Society listed C. elongatus as vulnerable, the status applied to a taxon that is in imminent danger of becoming threatened throughout all or a significant portion of its range (Jelks et al. 2008). Within the upper Midwest, the dace is extirpated from Iowa, listed as endangered in Canada, Michigan, and Indiana, and listed as a species of special concern in Wisconsin (Harlan and Speaker, 1956; Lyons et al., 2000; COSEWIC, 2007). Several populations of C. elongatus have disappeared from the upper Rock River basin in Dane County, Wisconsin (Lyons et al., 2000).

The continued occurrence of the redside dace in Illinois is largely dependent upon the integrity of the headwater habitats in Wisconsin that support reproductively viable populations. If the Wisconsin populations remain intact, one might expect the redside dace to periodically appear in Illinois waters. While it is possible that the specimens col¬ lected in East Fork Raccoon Creek (all juveniles) had been washed downstream from source populations in Wisconsin headwaters, it is also likely that C. elongatus occurs in extremely patchy, isolated schools that are easily missed even during extensive sampling.

ACKNOWLEDGEMENTS

This study was supported by funds provided by the Illinois Department of Natural Resources’ Illinois Wildlife Preservation Fund and by the Illinois Department of Transportation. Permission to collect in Wisconsin was given by Wisconsin Department of Natural Resources. Many families, especially the Williams family, kindly granted permission to sample the waterways on their property. B.D. Cheek, A.R. Kuhns, H. Sabaj, M.D. Sabaj, J.L. Sherwood, and A. Stultz assisted in the field. C.A. Mayer assisted in compiling records of vouchered specimens. J. Haas and R. Sauer shared accounts of collecting rare fishes. B. Zimmerman shared experiences with Clinostomus elongatus. S.M. Jaworski offered suggestions and assisted with map preparation. P. Willink (FMNH) and M. Pauers (MPM) generously provided access to specimens under their care.

LITERATURE CITED

Becker, G.C. 1983. Fishes of Wisconsin. University of Wisconsin Press, Madison. 1052 pp.

Burr, B.M., K.M. Cook, D.J. Eisenhour, K.R. Piller, W.J. Poly, R.W. Sauer, C.A. Taylor, E.R. Atwood, and G.L. Seegert. 1996. Selected Illinois fishes in jeopardy: new records and status evaluations. Transactions of the Illinois State Academy of Science 89:169-186.

Burr, B.M. and L.M. Page. 2009. Illinois fish communities: more than a century of change. Pp 147- 162 in C.A. Taylor, J.B. Taft, and C.E. Warwick, editors. Canaries in the catbird seat: the past, present, and future of biological resources in a changing environment. Illinois Natural History Survey Special Publication 30, Champaign. 306 pp.

Burr, B.M. and M.L. Warren, Jr. 1986. Status of the bluehead shiner (. Notropis hubbsi ) in Illinois. Transactions of the Illinois Academy of Science 79:129-136.

Burr, B.M., M.L. Warren, Jr., and K.S. Cummings. 1988. New distributional records of Illinois fishes with additions to the known fauna. Transactions of the Illinois State Academy Science 81:163-170.

Chick J.H., R.J. Maher, B.M. Burr, and M.R. Thomas. 2003. First black carp captured in US. Sci¬ ence 300:1876-1877.

151

Committee on the Status of Endangered Wildlife in Canada (COSEWIC). 2007. COSEWIC assess¬ ment and update status report on the redside dace Clinostomus elongatus in Canada. Committee on the Status of Endangered Wildlife in Canada, Ottawa, vii + 59 pp.

Fago, D. 1982. Distribution and relative abundance of fishes in Wisconsin, I. Greater Rock River Basin. Wisconsin Department Natural Resource Technical Bulletin 136:1-120.

Gilbert, C.R. 1980. Clinostomus elongatus (Kirtland), redside dace. Pp 148 in D.S. Lee, C.R. Gil¬ bert, C.H. Hocutt, R.E. Jenkins, D.E. McAllister, and J.R. Stauffer, Jr., editors. Atlas of North American freshwater fishes. North Carolina State Museum Natural History, Raleigh. 854 pp.

Harlan, J.R. and E.B. Speaker. 1956. Iowa fish and fishing. 3rd edition. Iowa State Conservation Commission, Des Moines. 377 pp.

Hiland, T.M. and W.J. Poly. 2000. The bleeding shiner, Luxilus zonatus, in Illinois (Cyprinidae). Transactions of the Illinois State Academy of Science 93:145-147.

Irons, K.S., M.A. McClelland, and M.A. Pegg. 2006. Expansion of round goby in the Illinois water¬ way. American Midland Naturalist 156:198-200.

Jelks, H.L., S.J. Walsh, N.M. Burkhead, S. Contreras-Balderas, E. Diaz-Pardo, D.A. Hendrickson, J. Lyons, N.E. Mandrak, F. McCormick, J.S. Nelson, S.P. Platania, B.A. Porter, C.B. Renaud, J.J. Schmitter-Soto, E.B. Taylor and M.L. Warren Jr. 2008. Conservation Status of Imperiled North American Freshwater and Diadromous Fishes. Fisheries 33:372-407.

Koster, W.J. 1939. Some phases of the life history and relationships of the cyprinid, Clinostomus elongatus. Copeia 1939:201-208.

Laird, C.A. and L.M. Page. 1996. Non-native fishes inhabiting the streams and lakes of Illinois. Illinois Natural History Survey Bulletin 35:1-51.

Lyons, J., P.A. Cochran, and D. Fago. 2000. Wisconsin fishes 2000: status and distribution. Univer¬ sity of Wisconsin Sea Grant Institute, Madison, Wisconsin. 87 pp.

Novinger, D.C. & T.G. Coon. 2000. Behavior and physiology of the redside dace, Clinostomus elongatus, a threatened species in Michigan, Environmental Biology of Fishes 57:315- 326.

Page, L.M. and B.M. Burr. 2011. A field guide to freshwater fishes of North America north of Mexico. 2nd Edition. The Peterson Field Guide Series, Houghton Mifflin Company, Boston, Massachusetts. 663 pp.

Page, L.M., K.S Cummings, C.A. Mayer, S.L. Post, and M.E. Retzer. 1992. Biologically significant Illinois streams. An evaluation of the streams of Illinois based on aquatic biodiversity. Illinois Natural History Survey, Center for Biodiversity Technical Report 1992(1). Champaign. 485 pp.

Poly, W.J. and A.K. Wilson. 1998. The fringed darter, Etheostoma crossopterum , in the Cache River Basin of Southern Illinois (Percidae: Subgenus Catonotus). Ohio Journal of Science 98:6- 9.

Retzer, M.E. 2005. Changes in the diversity of native fishes in seven basins in Illinois, USA. American Midland Naturalist 153:128-141.

Sabaj, M.H. 2000. Illinois status survey of the redside dace, Clinostomus elongatus, the newest addition to the state’s native fauna. Technical Report submitted to the Illinois Department of Natural Resources, Division of Natural Heritage. 12 pp + appendix.

Sabaj Perez, M.H. (editor). 2012. Standard symbolic codes for institutional resource collections in herpetology and ichthyology: an Online Reference. Verson 3.0 (23 February 2012). Electroni¬ cally accessible at http://www.asih.org/, American Society of Ichthyologists and Herpetologists, Washington, DC.

Smith, P.W. 1971. Illinois streams: a classification based on their fishes and an analysis of factors responsible for disappearance of native species. Illinois Natural History Survey Biological Notes 76:1-14.

Smith, P. W. 1979. The fishes of Illinois. University of Illinois Press, Urbana. 314 pp.

Steuck, M.J., S. Yess, J. Pitlo, A. Van Vooren, and J. Rasmussen. 2010. Distribution and relative abundance of upper Mississippi River fishes. Upper Mississippi River Conservation Committee, Onalaska, Wisconsin. 21 pp.

Stewart, J.G., V.B. Barko, D.B. Henry, D.P. Herzog, J.W. Ridings, A.F. Kelley, and J.E. Wallace. 2005. New records of the crystal darter ( Crystallaria asprella) in the middle Mississippi River. American Midland Naturalist 154:471-473.

Taylor, C.A. J.H. Knouft, and T.M. Hiland. 2001. Consequences of stream impoundment on fish communities in a small North American Stream. Regulated Rivers: Research and Management 17:687-698.

152

Tiemann, J.S., H.R. Dodd, N. Owens, and D.H. Wahl. 2007. Effects of lowhead dams on unionids in the Fox River, Illinois. Northeastern Naturalist 14:125-138.

Tiemann, J.S., M.E. Retzer, and B.L. Tiemann. 2004. Range expansion of the state endangered bigeye chub Hybopsis amblops (Rafinesque) in Illinois. Transactions of the Illinois State Acad¬ emy of Science 97:255-257.

Trautman, M.B. 1981. The fishes of Ohio with illustrated keys, rev. ed. The Ohio State University Press, Columbus. 782 pp.

Warren, M.L., Jr. and B.M. Burr. 1989. Distribution, abundance, and status of the cypress minnow, Hybognathus hayi, an endangered Illinois species. Natural Areas Journal 9:163-168.

Zimmerman, B.J. 2009. Microhabitat use by the redside dace ( Clinostomus elongatus ) in Ohio. Master’s thesis. Bowling Green State University, Bowling Green, Ohio. 28 pp.

153

BOOK REVIEW 201 2- #2

Side Channels. A Collection of Nature Writing and Memoir by Thomas V. Lerczak (201 1). 186 pp. Mill City Press, Minneapolis, MN. $15.95.

This book is an unusual hybrid: part birder’s field guide, part general natural history, and part memoir. As a study in contradictions we learn that the author grew up in urban Chi¬ cago and how he came to live in the open country near the Illinois River. We learn how, from an early age, he was drawn to nature, and especially to observation of birds. Follow¬ ing his ecological graduate studies Lerczak became a state biologist. In his duties as a staff member of the Nature Preserves Commission, he has come into close contact with many habitat types. Through his work and through his writing for Illinois Audubon Magazine, he has emerged as one of Illinois’ premier naturalists.

The volume is a collection of 30 chapters, fourteen of which originally appeared in Illi¬ nois Audubon magazine. Others are personal observations and reflections seen here for the first time. In the memoir-based chapters he notes that he is especially drawn to rivers. His descriptions of them, and their wildlife, are especially evocative.

Longtime Illinois residents will doubtless find much they did not already know. Lerczak always digs into the back story as he describes topics such as the habits of Great Blue Herons, eagles in winter refuges, the life of the six species of swallows along the Illinois River, the reasons why we have bur oaks in Mason County, or the territories of Red¬ headed Woodpeckers. We learn why hawks don’t glide over large lakes, and of the habi¬ tat needs of vultures vs. eagles. We learn why suppression of fires has the unintended consequence of changing the composition of oak forests.

Many chapters are essays on birds of this state. In part II, he travels more widely offering interesting observations of Colorado, Wisconsin, the North Cascades, the Great Lakes including Isle Royale, the Porcupine Mountains wilderness, and New York at Niagara Falls. He feels the contentment that comes from a lifetime of kinship with the earth’s community of life.

The sense of peace that one can feel in nature has led many of us to proceed down the road of environmental activism. Lerczak has stood at the entrance of this road but, in the end, rejected the politics and egos of agenda driven groups. This may puzzle those who are more politically engaged with environmental protection. But his reluctance overshad¬ ows the fact that, professionally, he has contributed a great deal to the actual preservation of thousands of acres of natural habitat within his home state. This is a major and endur¬ ing legacy that few are in a position to claim.

I note, in full disclosure, that Tom Lerczak was once a student in my Colorado field course. His sense of humor and earnest thoughtfulness were evident then as was the fact that he never went anywhere without his binoculars. It was clear in those days that his future career would bear watching.

Lerczak’s book displays an understated, companionable style as he introduces habits and habitats in a way that will appeal to novices as well as those more expert. Each chapter is

154

a good stand-alone read. This book should be read by any natural history buff, and by anyone looking for ideas of places to explore, both near and far.

Review by Richard C. Keating, Research Associate at the Missouri Botanical Garden and Professor Emeritus at Southern Illinois University - Edwardsville.

155

ADDITIONAL 2012 ISAS STUDENT GRANT AWARD RECIPIENTS

Kendall Annetti - University of Illinois - Urbana-Champaign

Avian Hematozoa of Illinois and Their Impacts on Health

ADDITIONAL 2012 ANNUAL MEETING STUDENT PRESENTATION AWARDS

SCIENCE, MATHEMATICS, & TECHNOLOGY EDUCATION DIVISION

Oral Presentations

1st Place: Gerry Mestek - Aurora University

A Study on the Arboreal Flora and the Environmental Conditions in a Bottomland Forest Site, Duck Creek Nature Trail,

Silver Springs State Park, Kendall County, Illinois

New York Botanical Garden Library

3 51

85 00342 6085

INFORMATION FOR CONTRIBUTORS

Submission Information

1. Beginning 2013 Transactions will be moving to an online format for submission and publication of manuscripts. As of April 1, 2013, manuscripts can be submitted online at http://www.ilacadofsci.com. You may also mail hardcopy manuscripts to the Transactions Editor but must include an electronic copy. The Editor’s contact information is Dr. Tere North; email terenl956@gmail.com; postal mail to 1713 W. Adams Rd., Macomb IL 61455.

2. Submission of a manuscript to the Editor will be held to imply that it has not been previously published except in Abstract form and, if accepted, will not be published elsewhere in the same form without written consent of the Editor.

3. Manuscript format should follow a form appropriate to the specific discipline: most commonly, e.g., Abstract, Introduction, Materials and Methods, Results, Discussion, and Summary.

4. Tables and figures must each be numbered consecutively in the order they are referred to in the text. At manuscript submission, authors may include tables/figures either at the end of or in the body of the manuscript. At publication all tables and figures will be incorporated into the body of the manuscript as close to first reference as possible.

5. Genus and species must be given for all organisms used in the investigation. Common names are also recommended.

6. In the text, references should be quoted by the author’s name and date, e.g., (Smith, 1960). Footnotes are to be avoided.

7. At the end of the manuscript, full references, must be listed in alphabetical order and include the names of the authors, date of publication, full title of the paper, title of the journal, volume number, and page numbers. References to books must include the number of the edition, publisher, and place of publication.

8. Authors are encouraged to submit names and addresses of potential content reviewers to add to the Transactions reviewers database. All reviews are blind to the author to maintain the confidentiality of the reviewer.

Acceptance Information

9. Each manuscript will be reviewed by no less than two anonymous reviewers.

10. For manuscripts deemed acceptable for publication, authors must address the reviewers comments and provide a revised copy to the Editor.

Publication Information

1 1 . Upon final acceptance of a manuscript for publication, the author shall provide the Editor with electronic copy (e.g., floppy, CD-ROM, or electronic e-mail attachment) of the final version of the manuscript including all text and tables.

12. All graphs, charts, and diagrams should likewise be provided in suitable electronic format (JPEG, TIFF, PICT, etc.), with a minimum 300 dpi resolution for images. For copy not available electronically, high quality, glossy originals must be provided for direct scanning.

13. Pre-press proofs will be sent to the author or designee for review prior to publication. Corrections to the page proofs are restricted to minor revision and due to printer error. Other alterations will be made at cost and charged to the author.

14. Academy member page charges are $50.00 per page. The charge for non-members is doubled.

15. Color figures, etc. may be used for the online publication format. When a year end hard copy of all manuscripts is printed for limited distribution it will be in grey-scale format. An electronic PDF copy will provided to the corresponding author following receipt of page charges and publication.

16. Credit card payment will be available for manuscript publication via secure PayPal beginning April 1 , 2013, at http://www.ilacadofsci.com.

Transactions of the Illinois State Academy of Science Volume 105 (2012)

CONTENTS

Academy of Science Editorial Information . inside front cover

Debut of Manuscript Submission and Publication Announced . 77

Botany Papers

Petersen, Chris E., Sharon M. Bauzys, Felicia A. Speranske, and Barbara A. Petersen The Varying Phenology and Growth Patterns of Baptisia bracteata (Fabaceae) in Reconstructed Prairie . 79

Environmental Science Papers

McClain, William E., John E. Ebinger, Roger Jansen, and Gordon C. Tucker

Vascular Flora of Capel Glacial Drift Hill Prairie Natural Area, Shelby County,

Illinois . 85

Bollinger, Eric K. and Brian Conklin

The Effects of Beaver-created Wetlands on Surface Water Quality of Lotic Habitats

in Northern Illinois . 93

Capparella, Angelo P., Todd Springer, and Lauren E. Brown

Discovery of New Localities for the Threatened Kirtland’s Snake ( Clonophis

kirtlandii ) in Central Illinois . 101

Collins, Kathleen A. and David Joseph Horn

The Role of Oil Content and Size in Seed Selection by Wild Birds . 107

Nielsen, Clayton K. and Susan E. Cooper

Comparison of Methods to Detect Mesocarnivores in Southern Illinois . 1 19

Nixon, Charles M., Philip C. Mankin, Dwayne R. Etter, Lonnie P. Hansen,Paul A. Brewer,

James E. Chelsvig, Terry L. Esker, and Joseph B. Sullivan

Selection of Parturition Sites by Migrating and Dispersing Female White-tailed

Deer in Illinois . 129

Tiemann, Jeremy S. and Mark H. Sabaj Perez

Illinois Status Survey of the Redside Dace Clinostomus elongatus: The Newest Addition to the State's Native Fish Fauna . 145

Book Reviews

Keating , Richard C.

Side Channels. A Collection of Nature Writing and Memoir by Thomas V. Lerczak.153

Announcements

Additional 2012 Student Research Awards and Grants . 155

Additional 2012 Annual Meeting Student Awards . 155

Contributor Information

inside back cover