AN ABSTRACT OF THE THESIS OF MARCUS C. ABSTON, for the Master of Science degree in ZOOLOGY, presented on 18 April 2017, at Southern Illinois University Carbondale. TITLE: EFFECTS OF OLFACTORY ENRICHMENT ON AFRICAN CHEETAHS (ACINONYX JUBATUS) MAJOR PROFESSOR: Dr. Carey Krajewski Environmental enrichment has been an essential part of felid husbandry in zoos, serving to reduce both physiological and psychological stress. Olfactory enrichment is one of many interventions used to prevent stereotypic behavior caused by stress in felids. However, little research has been done on this practice. The purpose of this study was to compare behavioral responses of four captive African cheetahs to six types of commercial fragrances used as olfactory enrichment stimuli. Three of these fragrances are marketed as “men’s cologne” and three as “women’s perfume”. The fragrances were also categorized as musky, spice, and floral types. Behavioral responses were recorded by live observations during 36 bouts at the St. Louis Zoo. There was a significant difference between cheetahs’ interaction time with male and female fragrances; they seemed to prefer male colognes. However, there was no significant difference in interaction time among fragrance types. Engagement behaviors varied among individuals (e.g. sniffing, scent rolling, pawing), but were similar to those reported by previous authors. These findings suggest that commercial fragrances may be a useful option for African cheetah olfactory enrichment.

Swede midge, Contarinia nasturtii Kieffer (Diptera: Cecidomyiidae), is an invasive pest causing marketable losses on Brassica crops in the Northeastern United States and throughout southern Canada. Heading brassicas, like cauliflower and broccoli, are particularly susceptible because larvae feed concealed inside meristematic tissues of the plant, where head formation occurs. Our work details the development of a sustainable, affordable pest management tactic for swede midge – plant derived repellents. First, it was necessary to establish both a damage and marketability threshold for swede midge, so we developed a technique to manipulate larval density of swede midge on cauliflower, We asked: (1) What is the swede midge damage threshold? (2) How many swede midge larvae can render cauliflower crowns unmarketable? and (3) Does the age of cauliflower at infestation influence the severity of damage? We found that even a single larva causes mild twisting and scarring rendering cauliflower unmarketable 52% of the time, with more larvae causing more severe damage and additional losses, regardless of cauliflower age at infestation. Repellency is an important management approach to consider for swede midge. Since the host range of specialist insects appears constrained by plant phylogeny, we hypothesized that odors from less phylogenetically related plants would be more repellent to swede midge. To test our hypothesis, we performed no-choice and choice biological assays, asking: (1) How do essential oils from different plant species influence midge densities on broccoli? (2) What is the relationship between phylogenetic distance of non-host odors and larval densities on broccoli? Biological assays identified multiple essential oils that reduced larval densities, and phylogenetic analyses showed that less related plants were more effective. In addition to the biological assays, we tested 15 essential oils for their ability to repel gravid females from broccoli tissue in y-tube olfactometer assays. While most of the essential oils reduced the frequency at which females chose host plant meristems, wintergreen, thyme, lemongrass, eucalyptus lemon, garlic, cinnamon, and star anise were most effective. Additionally, we used chemical fingerprints (physical/chemical properties) from PubChem to compare the essential oil volatile compounds and develop an index for their similarity. We found that physicochemical similarity was predictive of repellency. Finally, for repellency to be an effective, long-term strategy, it was important to consider how and whether the repellent response of midges changes over time or previous experience. In our final chapter, we performed electroantennography trials testing how previous experience with garlic or eucalyptus lemon odor for one or 10 s influences the neurophysiological response of swede midge to host (broccoli) or non-host (garlic or eucalyptus lemon) odors. We asked: (1) Does previous experience with garlic or eucalyptus lemon influence the physiological response of swede midge to host or non-host odors? (2) Does the time of previous exposure to non-host odors influence their physiological response to host or non-host odors? Our findings show that swede midge, after 10 s of exposure to either repellent, was more responsive to repellents than host compounds, suggesting that the effectiveness of repellents will not diminish over time.

Elasmobranchs (sharks, skates and rays) possess a sophisticated array of sensory systems that are, undoubtedly, of great importance to their survival. Representing the earliest group of extant jawed vertebrates, detailed study of elasmobranch sensory biology can provide much-needed information on the evolution of vertebrate sensory systems. Some sensory modalities have been studied in detail in several species, but few studies have examined and compared multiple sensory systems within a particular genus. By examining the morphology and physiology of the different sensory systems, correlations can be made within both an ecological and a phylogenetic context. The primary advantage of studying the sensory systems of closely related species is that any differences between them are more likely to reflect functional ecological adaptations rather than the effects of phylogenetic separation. Wobbegongs sharks (Orectolobidae) are a distinctive group of benthic sharks that are characterised by a highly patterned, dorso-ventrally compressed body. Wobbegongs are ambush predators that employ a unique ‘sit and wait’ strategy. Their morphologically distinct body shape, sedentary lifestyle and mode of predation suggest that wobbegong sharks may differ from other elasmobranchs in how they employ their different sensory systems. In this study, four wobbegong species that vary in life-history and/or habitat were examined: the Western wobbegong, Orectolobus hutchinsi, the spotted wobbegong, O. maculatus, the ornate wobbegong, O. ornatus and the dwarf spotted wobbegong, O. parvimaculatus. Vision and olfaction were assessed in all four species. Detailed assessment of electroreception and mechanoreception (lateral line) was conducted only for O. maculatus and O. ornatus. Morphology, physiology and molecular genetics were examined in the visual system, and morphological assessment was conducted for the olfactory, electroreceptive and mechanosensory lateral line systems. The retinae of all four wobbegong species are duplex; rod and cone photoreceptors can be distinguished easily on the basis of morphology. The wavelength of maximum absorbance (λmax) of the rod visual pigment is 496 nm in O. hutchinsi, 484 nm in O. maculatus, 498 nm in O. ornatus and 494 nm in O. parvimaculatus. Absorbance spectra of cone visual pigments were only obtained from O. maculatus and O. ornatus. Only one spectral type of cone was measured in each species, with max values at 553 nm and 560 nm, respectively. Partial sequences were obtained for the rh1 opsin gene in all four species, and for the lws opsin gene in every species except O. parvimaculatus. The apparent presence of only one cone pigment raises the possibility that wobbegongs do not have colour vision. The topographic distribution of cells within the ganglion cell layer of Orectolobus hutchinsi show a weakly elongated central visual streak of increased cell density, mediating a higher spatial resolving power of 2.06 cycles deg-1 in the frontal visual field. Retinal topography of O. maculatus and O. parvimaculatus are similar, with both possessing a dorsal horizontal streak facilitating increased spatial resolving power in the lower visual field. Orectolobus parvimaculatus also possesses an area of increased cell density in the naso-ventral region of the retina mediating acute vision in the upper caudal region of the visual field. Spatial resolving power reaches 3.51 cycles deg-1 and 3.91 cycles deg-1 in O. maculatus and O. parvimaculatus, respectively. The topographical variation in retinal sampling indicates that different regions of the visual field are relatively more important and may reflect interspecific differences in behaviour and habitat. The mean number of lamellae in the olfactory rosette is 47.0 for Orectolobus hutchinsi, 48.7 for O. maculatus, 40.7 for O. ornatus and 55.7 for O. parvimaculatus. Olfactory sensory epithelial surface area is comparable in O. hutchinsi, O. maculatus and O. ornatus, while O. parvimaculatus has a significantly larger surface area, relative to body size, compared to the other three species. Olfaction appears to be relatively more important in O. parvimaculatus, especially during low light conditions, when vision is limited. The distribution of ampullary electroreceptive pores and mechanosensory lateral line pores (pored and non-pored canals) is almost entirely concentrated on the dorsal region of the head in both O. maculatus and O. ornatus. This suggests that both sensory systems are well-adapted and specialised to detect prey swimming overhead when the wobbegong is sitting motionless, thereby facilitating its unique, predatory, “lie-in-wait’ ambush strategy. Orectolobus hutchinsi and O. ornatus appear to be well-suited to both diurnal and nocturnal activities, whereas O. maculatus and O. parvimaculatus are probably most active under low light conditions. Sensory system information inferred from this study correlates well with what is known of the diet and habitat of the four wobbegong species examined. Therefore, in the absence of other biological data, sensory neurobiological approaches can be used to predict such bio-ecological factors as predatory strategy, habitat preference, and behaviour. Electrophysiology and behavioural approaches will provide major advances in future studies in order to understand how each of the different senses is integrated at both peripheral and central levels and how such studies are vital to our understanding of evolutionary and ecological processes.

Les interactions perceptuelles entre différentes modalités sensorielles affectent la perception des aliments. Si ce phénomène a été largement étudié entre des stimuli olfactifs et gustatifs en solution aqueuse, peu d'études ont été menées avec des produits existants dans le commerce. Similairement, peu de travaux sont dédiés à l'étude des interactions multi-sensorielles impliquant l'olfaction, le goût et la perception tactile en bouche et à l'étude des perceptions dites complexes, c'est-à-dire qui impliquent plusieurs modalités sensorielles. Le premier objectif de ma thèse était l'exploration des mécanismes sous-jacents aux interactions perceptuelles existant entre l'olfaction et la gustation durant la consommation de produits réels, et en solution avec des stimuli olfactifs à une concentration infraliminaire. Cette approche a ensuite été étendue aux perceptions olfactive, gustative et tactile (en bouche) et à la perception complexe "rafraîchissante". Il a été mis en évidence que la familiarité d'un produit et la stratégie d'attention durant l'exposition affectent de façon critique les interactions perceptuelles. Pour la première fois il a été démontré qu'une concentration infraliminaire d'odorant associé au goût sucré (fraise) augmente la perception sucrée d'une solution de sucrose. La multiplicité des interactions sensorielles présente dans un milieu alimentaire complexe a également été mis en évidence à un niveau bimodal (ex: entre la perception amer et le froid) et tri-modal (ex: entre la perception olfactive, sucrée et froide). Finalement il été démontré que la perception rafraîchissante est construite sur la base d'une combinaison de déterminants sensoriels, d'habitudes alimentaires ainsi que de facteurs hédoniques et psychophysiologiques tels que l'énergie mental. Pour conclure, les connaissances acquises par ce travail soulèvent d'autres interrogations notamment à propos des mécanismes neuronaux sous-tendant la mémorisation des associations perceptuelles et des conditions requises en termes de durée et de fréquence d'exposition pour la mise en place de ces interactions.

<p>Olfactory function is impaired in aging and dementia. The general aim of this thesis was to identify variables that predict olfactory function and dysfunction (assessed with an odor identification test) in middle-aged and elderly adults. The thesis investigated whether odor identification performance was associated with demographic variables, cognitive function, the ApoE gene, dementia, and other health-related variables. The ApoE-ε4 allele is associated with cognitive impairment and Alzheimer’s disease, the most common form of dementia. The studies included in this thesis used data from the Betula study, a large-scale, population-based prospective study on aging, memory, and health. Study 1 investigated demographic and cognitive predictors of odor identification ability in non-demented participants. The results showed that younger age, female sex, and high education contributed to better odor identification ability. Cognitive speed and vocabulary had a small additional influence. Study 2 included information about ApoE genotypes, dementia and other health-related variables. The results indicated that the ApoE-ε4 allele was associated with odor identification impairment among the elderly, but not middle-aged adults. Participants who were demented at the time of testing or became demented within five years after testing exhibited olfactory impairments. Interestingly, the age-related olfactory impairment in ε4-carriers was independent of clinical dementia within five years. In Study 3, decline in global cognitive status over a five-year test-retest interval was predicted in a sample of elderly participants. The major result was a three-way interaction reflecting that odor identification impairment, old age, in combination with the ε4 allele predicted a larger cognitive decline. However, odor identification impairment did not predict cognitive change in elderly who were non-carriers of the ε4 allele. Overall, the results indicate that odor identification impairment in elderly is related to ApoE-ε4, cognitive decline, and clinical and pre-clinical stages of dementia. Theoretical and practical implications of the results are discussed. Furthermore, it is proposed that in order to effectively predict clinical dementia or cognitive decline from olfactory assessment in the elderly, variables that mediate (e.g. neuropathology) or moderate (e.g. age) the associations between olfactory function, the ε4 allele, and dementia need to be further evaluated, preferably in studies using longitudinal assessment.</p>

<p>In this study I assessed the effect of objects and odors as environmental enrichment for two captive snow leopards (Uncia uncia) at Kolmården Wildlife Park. Five odors (lavender, lemon balm, cumin, cinnamon and catnip) were used to impregnate four different types of enrichment objects (boomer balls®, tennis balls, ropes, logs). During test sessions, one odorized and one non-odorized enrichment object of the same type were introduced in the snow leopards’ outdoor enclosure. The behavior, activity and location of the snow leopards were recorded and compared to their behavior during baseline sessions. During the test sessions I recorded the number and types of interactions with the enrichment objects. Both snow leopards interacted more often with odorized than with non-odorized enrichment objects. The number of interactions differed markedly between the types of enrichment objects and between the different odors. Both snow leopards interacted most often with boomer balls® and least often with logs. They interacted most often with cinnamon and least often with lemon balm (Brahma) and catnip (Binu). The results suggest that both the type of object and the odor play a role in capturing the interest of the snow leopards. The snow leopards behavioral diversity increased during the study and they could increase their performance of species-specific behavior. I did not see any indications of habituation during the testing period. The results indicate that enrichment objects impregnated with odors can be an effective environmental enrichment for captive snow leopards.</p>

<p>Odor discrimination ability was tested in four female South African fur seals (<em>Arctocephalus pusillus pusillus</em>) using a food-rewarded two-choice instrumental conditioning paradigm. The seals’ ability to distinguish between members of homologous series of aliphatic ketones (2-butanone to 2-heptanone) and alcohols (1-butanol to 1-heptanol) was assessed. The results showed that three out of four seals successfully discriminated between all of their stimulus combinations in both classes of odorants. One seal succeeded to reach the discrimination criterion with all 2-ketones but failed with all 1-alcohols.</p><p>No significant correlation between odor discrimination performance and structural similarity of the odorants in terms of differences in carbon chain length was found in either of the two chemical classes. Furthermore, it was found that the 2-ketones were significantly better discriminated than the 1-alcohols. The fact that both classes of odorants are known to be present in the natural environment of seals provides a possible explanation as to why most of the seals were able to successfully discriminate between them. The results of the present study support the notion that the sense of smell may play an important role in behavioral contexts such as social communication, foraging and reproductive behavior of fur seals.</p>

No description available.

<p>In this thesis lateralization of olfactory functions was investigated by both behavioral and electrophysiological assessment, the latter with the olfactory event-related potential (OERP) technique. The olfactory sense is primarily ipsilateral in that a stimulus that is presented to one nostril is initially processed in the same hemisphere. This makes it possible to observe differences between stimulated nostrils as an indication of hemispheric difference. Study I explored differences in olfactory cognitive functions with respect to side of rhinal stimulation and demonstrated that familiarity ratings are higher at right- compared to left-nostril stimulation. No differences were found in episodic recognition memory or free identification, possibly reflecting inter-hemispheric interactions in higher cognitive functions. Effects of repetition priming were present in odor identification and tended to be more pronounced when tested via left nostril. Study II further investigated the effect of previous exposure in odor identification by a different experimental set-up, and demonstrated effects of repetition priming when tested via left- but not right-nostril stimulation. This finding indicates the importance of reconsidering possible sequential effects in olfactory research. Study III examined methodological aspects of an OERP protocol with respect to stimulus duration, which was used in Study IV. No differences in amplitudes or latencies where found between the stimulus durations of 150, 200 and 250 ms, suggesting the commonly used duration of 200 ms in a standard protocol. Study IV investigated laterality effects in OERPs with respect to side of stimulation and electrode site. The results showed consistent amplitudes and latencies regardless of rhinal side of stimulation. Larger amplitudes were demonstrated on left hemisphere and midline compared to right hemisphere, possibly explained by smaller N1/P2 amplitudes at the right-hemisphere sites at left-nostril stimulation. Apart from a proposed OERP protocol, the findings support the notions of a right-hemisphere predominance in processes related to olfactory perception and indicate, in accordance with other findings, a left-side advantage in conceptual repetition priming.</p>

It has been proposed that the various 14-3-3 isotypes and isoforms present in all eukaryotes are largely functionally equivalent. However, this is not consistent with the conservation of multiple isoforms and isotypes, especially in vertebrates with seven 14-3-3 encoding genes and nine isotypes. The hypothesis tested in this thesis is that both isoform-specific and overlapping functions are likely mediated through tissue specific expression, colocalization and dimerization of 14-3-3 proteins occur in vivo. Drosophila melanogaster was selected because it offers a simple, but representative system to study these proteins functionally. This thesis focuses primarily on D14-3-3?, although the expression pattern and phenotypes associated with all three Drosophila 14-3-3s were determined. I first determined the expression pattern of the three different 14-3-3 isotypes (leoI, leoII and D14-3-3?) and described developmental phenotypes associated with mutations in 14-3-3 isotypes in Drosophila. I found that there is partial redundancy with respect to lethality. Both LEO and D14-3-3? appear required for normal germ-line and somatic gonadal development. However, they do not appear to be functionally equivalent with respect to this phenotype since LEO is unable to compensate for the loss of D14-3-3?. I also determined that D14-3-3? mutants have unique phenotypes including deficits in adult cross-vein formation and rapid habituation to olfactory and footshock stimuli. To further understand the unique role that D14-3-3? plays in the adult CNS, I mapped the areas in the brain involved in olfactory and footshock habituation. I found that although the mushroom bodies (MBs) are necessary to inhibit premature habituation such as that exhibited by D14-3-3? mutants, D14-3-3? expression specifically in the MBs is not sufficient to rescue premature habituation. Although the loss of either LEO or D14-3-3? appears to cause a deficit in olfactory associative learning, premature habituation is the cause of the deficit seen in D14-3-3? mutants. As leo mutants do not exhibit a premature habituation phenotype, it appears that within the MBs LEO and D14-3-3? are not functionally equivalent. Therefore, the data supports the hypothesis that 14-3-3s have functional specificity and redundancy likely to represent use of homo and heterodimers in different processes within the tissues of an organism.