The mission of the Department of Biology at Indiana State University is to nurture the academic potential of every student, advance knowledge through quality research, and serve the university and broader communities. Through experiential learning both within and outside the classroom, we motivate our students to become critically thinking, informed citizens with a heightened interest in science. Through one-on-one mentoring, we engage both undergraduate and graduate students in the excitement and challenge of original research. Through research and outreach education, we make valuable contributions at both local and national scales to the scientific community and the well-being of the general public.

Recent Submissions

  • ACOUSTIC COMMUNICATION IN THE TEMPERATE TREEFROGS PSEUDACRIS CRUCIFER AND ACRIS CREPITANS

    Keating, John (Indiana State University, 2014-08)
    Spring peepers and cricket frogs produce advertisement calls to attract females. As ectotherms their body temperatures are greatly affected by ambient air temperature. Some characteristics of their advertisement calls are correlated with temperature. I analyzed advertisement calls of both species recorded in western-central Indiana. I compared call characteristic correlations with temperature found in our populations to those in populations in other geographic areas and found similar trends throughout the range of both species. Secondly, I examined aggressive calls in the spring peepers. Aggressive calls are used in male-male interactions, and in the spring peeper are a distinct call type different from advertisement calls in two characteristics. I used a habituation-discrimination protocol to test which of the two call characteristics, that differ between advertisement and aggressive calls, males use to distinguish advertisement and aggressive calls. I found that males responded with intermediate aggression to calls that only differ from advertisement in one of the two characteristics and responded with the most aggression to calls that differed from advertisement calls in both characteristics.
  • FACTORS AFFECTING THE PROBABILITY OF ACOUSTIC DETECTION AND SITE OCCUPANCY OF BATS IN CENTRAL INDIANA

    Kaiser, Zachary David Epping (Indiana State University, 2014-05)
    Documenting the presence of rare bat species can be difficult. The current summer survey protocol for the federally endangered Indiana bat (Myotis sodalis) requires passive acoustic sampling with directional microphones (e.g., Anabats), but there are still questions about best practices for choosing survey sites and appropriate detector models. Indiana bats are capable of foraging in an array of cover types, including structurally-complex, interior forests. Further, data acquisition among different commercially available bat detectors is likely highly variable, due to the use of proprietary microphones with different frequency responses, sensitivities, and directionality. We paired omnidirectional Wildlife Acoustic SM2BAT+ (SM2) and directional Titley Scientific Anabat SD2 (Anabat) detectors at 71 random points near Indianapolis, Indiana from May-August 2012-2013 to compare data acquisition by phonic group (low, mid, Myotis) and to determine what factors affect probability of detection and site occupancy for Indiana bats when sampling with acoustics near an active maternity colony (0.20-8.39 km away). Weatherproofing for Anabat microphones was 45° angle PVC tubes and for SM2 microphones was their foam shielding; microphones were paired at 2 m and 5 m heights. Habitat and landscape covariates were measured in the field or via ArcGIS. We adjusted file parameters to make SM2 and Anabat data comparable. Files were identified using Bat Call ID software, with visual inspection of Indiana bat calls. The effects of detector type, phonic group, height, and their interactions on mean files recorded per site were assessed using generalized estimating equations and LSD pairwise comparisons. We reduced probability of detection (p) and site occupancy (ψ) model covariates with Pearson’s correlation and PCA. We used Presence 6.1 software and Akaike’s Information Criteria to assess models for p and ψ. Anabats and SM2s did not perform equally. Anabats recorded more low and midrange files, but fewer Myotis files per site than SM2s. When comparing the same model of detectors, deployment height did not impact data acquisition. Weatherproofing may limit the ability of Anabats to record Myotis, but Anabat microphones may have greater detection ranges for low and midrange bats. Indiana bat detections were low for both detector types, representing only 4.4% of identifiable bat files recorded by SM2s. We detected Indiana bats at 43.7% of sampled sites and on 31.4% of detector-nights; detectability increased as “forest closure” and mean nightly temperature increased, likely due to reduced clutter and increased bat activity, respectively. Proximity to colony trees and specific cover types generally did not affect occupancy, suggesting that Indiana bats use a variety of cover types in this landscape. Omnidirectional SMX-US microphones may be more appropriate for Indiana bat surveys than directional Anabat microphones. However, we conclude that 2 nights of passive acoustic sampling per site may be insufficient for reliably detecting this species when it is present. In turn, the use of acoustic monitoring as a means to document presence or probable absence should be reassessed.
  • PHYTOL DERIVED IMMUNOADJUVANTS AS OIL-IN-WATER MICRO-EMULSIONS FOR USE IN VACCINES

    Johnson, Dylan M. (Indiana State University, 2014-05)
    The main objective of this study is to evaluate an oil-in-water micro-emulsion of phytanol which we have named PHIS59 with multiple antigens: ovalbumin, a protein and microorganism that include a reference laboratory strain of methicillin susceptible Staphylococcus aureus (MSSA) and methicillin resistant S. aureus (MRSA). PHIS59 was compared with the previously described crude oil-in-water emulsion PHIS-01 and the similarly formulated squalene as an oil-in-water micro-emulsion based on the commercial product MF59 referred to here as NV59. Safety and efficacy of PHIS59 and NV59 was evaluated in a mouse and rabbit vaccine models. Determinations of safety were reported as the range of in vivo mouse LD50s for each compound. Vaccine efficacy was assessed in terms of antibody response titers and isotype profiles in sera of vaccinated animals. Vaccines consisted of ovalbumin or heat killed S. aureus adjuvanted with either PHIS59, NV59 or PHIS-01. Control animals received unadjuvanted controls. Currently, there is no approved MRSA vaccine. PHIS-01 has shown promise in preventing MRSA-associated mortality in a mouse model; this study provides cross species validation. PHIS59 is easier to handle than PHIS-01 because it does not bind the syringe plunger, can be preloaded into syringes and is a stable emulsion which allows consistent dose delivery. PHIS59 has a lower LD50 but is effective at lower doses giving a similar therapeutic index. This could be explained by increased bioavailability due to the micro-emulsion formulation. This is an improvement over PHIS-01 in that it is effective at lower doses. Finally, we tested a novel water soluble phytol derivative, sodium phytanyl sulfate and characterized a safe working dose range. Based on the doses determined through this study, sodium phytanyl sulfate (PHIS-SO4) is currently being studied as an adjuvant with an effective mouse dose of less than 1mg. Additionally, because it is soluble in water, it requires no emulsification for formulation in vaccines.
  • CHARACTERIZATION OF HYPOTHETICAL PROTEINS SAS0760 AND SAS1738 FROM “COMMUNITY ASSOCIATED STAPHYLOCOCCUS AUREUS” MSSA476

    Ramalinga, Anupama B. (Indiana State University, 2013-12)
    Staphylococcus aureus is known to cause a wide range of infections from simple, curable skin infections like carbuncles, furuncles, and impetigo to deadly infections such as bacteremia, osteomyelitis, endocarditis and post-operative infections. Recently, the emergence of multiple antibiotic resistances has posed a great challenge to therapeutics and infection management. Very early in therapeutics of Staphylococcus aureus infections, it acquired resistance to the penicillin, including methicillin [1], that was designed specifically for the penicillin resistant strains. These strains were designated methicillin resistant Staphylococcus aureus (MRSA). Originally, MRSA was only seen in hospital-associated infections (HA MRSA) [2-4], but recently, MRSA found in community settings (CA MRSA) are more invasive infections [5, 6] with greater genetic diversity and hence different antibiotic resistance patterns [4, 7]. MSSA476 is a CA MRSA, though evolutionarily, it is closely related to MRSA252, a HA MRSA [2, 8]. Hence we decided to identify hypothetical proteins from MSSA476. Disease management involves understanding the properties of bacteria in order to tackle the infection efficiently and finding new effective drugs to kill the pathogen. Further understanding of such unique, unexplored hypothetical proteins and their characterization with relation to virulence will help understand the increased virulence of CA MRSA.
  • Acoustic Communication in the Temperate Treefrogs Pseudacris Crucifer and Acris Crepitans

    Keating, John (2015-01-07)
    Spring peepers and cricket frogs produce advertisement calls to attract females. As ectotherms their body temperatures are greatly affected by ambient air temperature. Some characteristics of their advertisement calls are correlated with temperature. I analyzed advertisement calls of both species recorded in western-central Indiana. I compared call characteristic correlations with temperature found in our populations to those in populations in other geographic areas and found similar trends throughout the range of both species. Secondly, I examined aggressive calls in the spring peepers. Aggressive calls are used in male-male interactions, and in the spring peeper are a distinct call type different from advertisement calls in two characteristics. I used a habituation-discrimination protocol to test which of the two call characteristics, that differ between advertisement and aggressive calls, males use to distinguish advertisement and aggressive calls. I found that males responded with intermediate aggression to calls that only differ from advertisement in one of the two characteristics and responded with the most aggression to calls that differed from advertisement calls in both characteristics.
  • Using Stable Isotope Analysis to Study Altitudinal and Latitudinal Bat Migration

    Arias, Lily (2014-10-03)
    The general lack of knowledge on basic aspects of the biology of temperate and tropical bats, their low reproductive rates, and threats such as white nose syndrome, wind farms, and habitat loss, make them very susceptible to population declines.My research uses an innovative technique, the analysis of stable isotopes, to study the ecology of bat migration with the main goals of contributing significantly to the understanding of bat biology and assessing the conservation status and susceptibility of bats. In the first chapter,I measured the content of hydrogen isotopes in fur samples of migratory bat species killed at a wind farm in northern Indiana to determine their geographic origin.North American tree bats ( Lasiurus borealis, L. cinereus,and Lasionycteris noctivagans)are considered long distance migrants. In North America, peaks in bat mortality at wind farms occur between mid-July and mid-September. This period is associated with fall migration of bats from their summer (breeding) grounds to their wintering grounds. Thus, wind turbines may have serious negative effects on a strategic event in the life of bats by interrupting migratory connectivity and thereby imperiling the long-term persistence of migratory bat species at large scales.The analysis accurately predicted the known origin of control samples and estimated that non-control bats killed at the wind farm originated from several populations in the United States as well as in Canada. My results highlighted the threat of wind farms to local bat populations as well as to bats originating far from those farms, and emphasized the need for conservation policies across borders.High variation in stable hydrogen isotopes in migrant individuals of all 3 species was observed, suggesting that individuals or populations from a variety of regions pass through the wind farm. In the second chapter,I evaluated the triple-isotopic (hydrogen, carbon, and nitrogen) composition of the tissues of 7 bat species collected at 3 altitudes in the Central Andes of Peru,and the variation of these isotopes across an altitudinal gradient,the application of isotope analysis to migration studies, and trophic effect. Previous studies had demonstrated that iv hydrogen isotopes were a reliable tool to track altitudinal movements of birds, and there was evidence from soil and plant studies that nitrogen and carbon isotopes could serve the same purpose. However, studies focused on bats were lacking. Hydrogen and nitrogen isotopes in the sanguinivorous control were found to be enriched relative to those of the syntopic frugivores.Carbon isotopes in the sanguinivorous bat were depleted when compared to frugivores.Differences in hydrogen found between trophic groups are the first reported for the species studied and support results found elsewhere in the Neotropics.My results demonstrated that, in spite of the wide array of physiological and environmental factors producing temporal and spatial variation, the analysis of hydrogen isotopes is a promising tool to study altitudinal movements of bats when used over long distances. Neither stable isotopes of nitrogen or carbon appear to be reliable to track movements along short gradients such as those along mountains. The contrast of these findings with the results of previous studies suggests that isotopic gradients may be specific to given taxon and localities.My results contributed to the understanding of bat movement patterns and therefore to assessing their sensitivity to potential threats such as habitat loss and connectivity.
  • Diurnal and nocturnal avian antipredator behavior in thermally challenging conditions

    Carr, Jennie M. (2014-03-18)
    Diurnal avian antipredator behavior has been the focus of much past research, yet the influence of the thermal environment on such behaviors is often overlooked. Far less is known about nocturnal avian antipredator behavior, including how these behaviors are influenced by challenging thermal environments. The first portion of my research focused on how the thermal environment influences the diurnal antipredator behavior of wintering birds while (i) exposed to high wind speeds, (ii) foraging in sunlit and shaded microhabitats, and (iii) when using thermoregulatory postures to conserve body heat. In addition to increasing convective heat loss, high wind speeds increase the prevalence of background movements in the environment. My research demonstrated that wintering sparrows exposed to a moving stimulus are less likely to flush to cover on windy days than on calm days, suggesting that wind-driven visual noise may interfere with predator detection. Predator detection may also vary when feeding in sunlight and shade, and the thermal benefits of foraging in direct sunlight on cold winter days may also play an important role in dictating microhabitat choice. Regardless of the thermal benefits of foraging in sunlight, wintering sparrows preferred to feed in shaded microhabitats even at ambient temperatures well below thermoneutrality. However, these birds foraged in sunlight more frequently as ambient temperatures fell, suggesting a trade-off between thermoregulation (solar input) and predation risk. Additional evidence of such a thermoregulation-predation trade-off was evident in the use of heat-conserving thermoregulatory postures by wintering sparrows. Fluffing the feathers or standing on one foot will reduce the amount of heat lost to the environment. However, such postures slow take-off time and likely result in an increase in predation risk. As such, these risky postures were only used when feeding at relatively low ambient temperatures and when near protective cover. In general, these results indicate that characteristics of the thermal environment play an important role in dictating diurnal antipredator behavior. To address how the thermal environment influences nocturnal avian antipredator behavior, I examined the predation-related costs of using energy-saving nocturnal hypothermia. Many species of birds reduce their nighttime body temperature, thus reducing metabolic rate and conserving energy. Such drops in body temperature may be quite substantial and likely influence a bird’s ability to respond to a potential threat during the night. To examine the potential costs of hypothermia, I conducted nocturnal flight tests on hypothermic mourning doves (Zenaida macroura). In general, doves that cooled by more than 5 °C flew poorly or were unable to fly, but were able to fly well once re-warmed to near their normal daytime body temperatures. Thus, low body temperatures during energy-saving hypothermia likely result in an increase in the risk of nocturnal predation. Nocturnal antipredator behavior was also examined in ruby-throated hummingbirds (Archilochus colubris). These hummingbirds frequently use nocturnal torpor (i.e., deep hypothermia), with significant reductions in body temperature and corresponding inability to respond behaviorally to external stimuli. Although hummingbirds altered torpor use seasonally and over the course of the observation period, hummingbirds did not consistently reduce their use of torpor following an experimental increase in perceived predation risk. Thus, although hypothermia is behaviorally costly, further studies are needed to clarify the role of predation on nocturnal behavior in birds.
  • Evaluation of an Avian Radar System

    Gerringer, Michael B. (2014-03-18)
    The problem of bird strikes in aviation is becoming an increasing threat, both to aircraft and to human safety. Management efforts have reduced wildlife hazards below 500 feet and within the immediate airport environment, but traditional methods of monitoring bird activity are limited to an observer’s field of view. Avian radar systems could potentially be useful in monitoring bird activity at great distances from the airport, at higher altitudes, and at night (Dolbeer 2006), but little work has been done to validate the tracking capabilities of avian radar systems. Thus, the goal of this research is to evaluate the detection and tracking abilities of the Merlin Avian Radar System provided by DeTect Inc. Radio-controlled (RC) aircraft flights were used to systematically test the tracking abilities of the Merlin System with respect to distance and altitude. Transits by free-flying birds provided an equally important test of the Merlin System, allowing for the assessment of tracking performance as influenced by flock size, altitude, and distance from the radar unit. Overall tracking performance regarding the RC aircraft and single large bird targets was poor across all study distances and altitudes. However, flocks of large birds such as geese and cranes were tracked well, even those several miles away from the radar unit. Given these results, avian radar could be a useful tool for monitoring bird flock activity at airports, but less so for single large bird targets such as thermalling raptors.
  • Summer Indiana Bat Ecology in the Southern Appalachians: An Investigation of Thermoregulation Strategies and Landscape Scale Roost Selection

    Hammond, Kristina (2014-03-18)
    In the southern Appalachians there are few data on the roost ecology of the federally endangered Indiana bat (Myotis sodalis). During 2008-2012, we investigated roosting ecology of the Indiana bat in ~280,000 ha in the Great Smoky Mountains National Park, Cherokee National Forest, and Nantahala National Forest in the southern Appalachians Mountains of Tennessee and North Carolina. We investigated 2 aspects of the Indiana bat’s roosting ecology: thermoregulation and the extrinsic factors that influence body temperature, and landscape-scale roost selection. To investigate thermoregulation of bats at roost, we used data gathered in 2012 from 6 female Indiana bats (5 adults and 1 juvenile) to examine how reproductive condition, group size, roost characteristics, air temperature, and barometric pressure related to body temperature of roosting bats. We found that air temperature was the primary factor correlated with bats’ body temperatures while at roost (P < 0.01), with few differences detected among reproductive classes in terms of thermoregulatory strategies. To understand how Indiana bats select roosts on a landscape-scale, we created a presence-only model through the program MaxENT using 76 known roost locations to identify areas important to summer roosting habitat within our study area and to identify important landscape-scale factors in habitat selection. The final model showed that Indiana bats selected roosts on the upper portion of ridges on south facing slopes in mixed pine-hardwood forests at elevations of 260-700 meters. Unfortunately, due to small sample size and the large effort required to fully investigate thermoregulation of Indiana bats in the southern Appalachians, we only were able run correlations with temperature data, and further investigation is needed to make concrete conclusions. However, the new advancements in resolution of landscape cover data and new programs in spatial modeling have enabled us to produce a large scale spatial model for identifying Indiana bat summer roosting habitat within our study area. Our findings have added to our understanding of Indiana bat roosting ecology, particularly in the southern Appalachian Mountains, and will aid land managers in effective management for this federally endangered species.
  • Levels of Selection in a Polymorphic Species

    Korody, Marisa L. (2013-09-05)
    Phenotype is affected by many factors, including but not limited to environment, conspecifics, and genetics. Evidence of phenotypic variation is everywhere, some of which is controlled solely by environment, and others that are fixed by genetics. Genetic polymorphisms are rare, but very useful for the study of selection and genetics. These genetic polymorphisms provide a phenotypic link to the underlying genetics and are even more useful when there are associated behavioral differences. I examine multiple levels of selection that are acting upon a polymorphic passerine, the white-throated sparrow (Zonotrichia albicollis). Males and females of this species occur in two morphs, white or tan, based upon the color of their crown strips. This plumage polymorphism is absolutely correlated with a complex chromosomal rearrangement on the second largest autosome. Within this dissertation I explore how climate needs to be addressed in ecological studies to fully understand the mechanisms behind variation. I explore whether sexual selection is acting within this species and the differences between the morphs through the use of Bateman Gradients. Darwin suggested that sex ratios influence sexual selection, but what about morph ratios? I examine the frequency variation of morphs within this species. Variation in morph production may be favored by a potential tradeoff between the number of males and the number of white offspring produced in a clutch that suggests greater costs associated with producing white morph individuals. Mendelian segregation is inconsistent in this species, and transmission distortion may contribute to morph ratio variation. I show that white male sperm varies in production from 0% - 100% white sperm/individual consistent with transmission distortion. Finally, candidate gene mapping was used to identify the genes sequestered in this rearrangement that may be responsible for the polymorphism and the evolution behind the rearrangement.
  • A morphological and cultural study of a species of Nigrospora

    Patrick, Mildred (2013-03-08)
    Not Available.
  • Characterization of Novel Extracellular Matrix (Ecm) Proteins (Mgp and Lumican) and Their Implications in Vascular Development, Angiogenesis, and Cancer

    Sharma, Bikram (2013-01-30)
    Extracellular matrix (ECM) constitutes a large component of our tissue structure. Primarily, ECM provides structural and adhesive support to our cells, but it also controls cellular signaling and behavior. Homeostasis of extracellular matrix composition and function is maintained by our body through a balanced synthesis, degradation and remodeling of ECM. However, under pathological conditions and genetic mutations, ECM homeostasis is disrupted due to deregulation in ECM synthesis, assembly, remodeling, and degradation. A number of diseases, including cardiovascular diseases and cancer, are found to occur due to alterations in ECM. Therefore, targeting ECM can be an attractive therapeutic approach to treat these diseases, and it requires our complete understanding of the ECM molecules and the molecular mechanism it employs in controlling cellular functions. To this end, this study is aimed at the characterization of two ECM proteins—Matrix Gla Protein (MGP) and Lumican—for their roles in vascular development, angiogenesis, and cancer. Findings from this study show that MGP is a critical ECM regulator that promotes angiogenic resolution by suppressing endothelial sprouting and stabilizing vascular lumen formation. In addition, MGP also inhibits tumor growth by inhibiting tumor angiogenesis. On the other hand, our findings show Lumican suppresses tumor growth and has anti-angiogenic activity in a context specific manner.
  • Cloning and Characterization of Hypothetical Exported Proteins from Community Associated Staphylococcus Aureus

    Kaur, Haninder (2013-01-30)
    Methicillin resistant Staphylococcus aureus (MRSA), a major cause of nosocomial infections, has acquired resistance to beta-lactam and other antibiotics. Recently, community associated MRSA (CA-MRSA) has developed independent of hospital associated MRSA (HA-MRSA). One of the major differences between the hospital and community strains is that the former is multi-resistant to antibiotics while the latter is not as resistant but is significantly more invasive. This increased invasiveness and the ability to cause life-threatening infections, even in immunocompetent individuals, makes CA-MRSA critically important as a public health problem. CA-MRSA is known to cause skin and soft tissue infections; bacteria interact with host skin cells and gain access to deeper tissues causing invasive infections. During this process the bacteria may secrete proteins that aid in the interaction with the host by adhering, invading or causing host cell death and lysis. To understand the virulence mechanisms involved in invasion, we investigated genes described as hypothetical proteins in MSSA476. The bioinformatics-selected proteins showed high probability of being secreted and most were unique to CA-MRSA. Our analysis showed 24 such genes. This study shows primer design for 15 of the genes (7 of the 24 had already been cloned in our laboratory). Using gateway cloning, the 15 genes were cloned into BL-21 expression clones. CA-MRSA’s are known for causing invasive skin infections. To further understand the involvement of our proteins of interest in invasion, human keratinocyte cell lines were used in a study of virulence and interaction with skin. To understand the involvement of our hypothetical secreted proteins, we investigated the mRNA expression level, using RT-qPCR and Livak method, of 20 hypothetical exported proteins in presence of human dermal keratinocyte cell line. Our investigation revealed two genes that showed increased mRNA expression in the presence of keratinocytes, which may be due to factors associated with keratinocytes that may have triggered increased mRNA expression. Keratinocytes are capable of forming cell-cell junctions and producing antimicrobial peptides and cytokines in response to microbes. The increased mRNA expression of two genes may be towards binding to junctions for invasion or may be expressed in response to antimicrobial peptides or cytokines.
  • A self improvement sheet for biology teachers in the secondary schools

    Roberts, Ernestine Winifred (2013-01-22)
    Not available

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