This thesis examines three factors that may influence the change in protein content and size of the brood food glands in honey bees. Effects on the mandibular gland, involved in the production of brood food and in royal jelly, have not been examined in relation to primer pheromones while effects on the hypopharyngeal glands, also involved in the production of brood food, have not been examined in relation to queen mandibular pheromone. This thesis provides preliminary insight into how these pheromones affect the extractable protein content of brood food glands. The first study in this thesis assessed the effects of brood pheromone (BP), queen mandibular pheromone (QMP), and pollen presence on the protein content of hypopharyngeal and mandibular glands of the honey bee. In this study, newly emerged bees were caged for 12 days in one of eight treatments: Queenless state: 1) control (no pollen + no pheromone), 2) pollen, 3) BP, 4) BP + pollen; Queenright state: 1) QMP, 2) QMP + pollen, 3) BP + QMP, 4) BP + QMP + pollen. This study indicated that regardless of pheromone treatment, the most influential factor on gland protein content and size was pollen. The second experiment examined effects of varying pollen dilution on hypopharyngeal and mandibular gland protein content, bee mass, and lipid content of the honey bee. In this experiment, newly emerged bees were caged for 7 days and fed one of five treatments: pollen, 1:1 pollen: cellulose (vol:vol), 1:2 pollen: cellulose (vol:vol); 1:3 pollen: cellulose (vol:vol), and cellulose. This study indicated that bees on the pollen diet were significantly greater than all other diluted diets in measurements of hypopharyngeal gland protein content, lipid content, and mass with significantly less consumption. However, mandibular gland protein content of bees on the pollen diet was significantly greater only from pure cellulose.

One lesser-known but universal post transcriptional modification carried out in yeast and higher eukaryotes is the methylation of mRNA, as mediated by the Ime4 protein and its orthologs. Ime4 protein is essential for sporulation in yeast cells and for viability of higher eukaryotic cells. The precise locations of the Ime4 protein and the functions of the methylated mRNA are still largely unknown. Whereas Ime4 protein is believed to be exclusively nuclear in higher eukaryotes, we have observed the yeast Ime4 protein in the nucleus, in the cytosol and within cytosolic particles. These observations suggest that Ime4 could be a shuttling RNA binding protein, playing roles in the cytosol as well as the nucleus. As a first step to examining this idea, we tested the hypothesis that the punctuate cytosolic particles formed by Ime4 are P bodies. P bodies are transient aggregates of proteins and RNAs that form as a result of stresses such as glucose deprivation. This experiment was carried out using fluorescence microscopy using Ime4 tagged with GFP (green fluorescent protein) and the known P -body proteins Edc3, tagged with mCherry. We expected that if the proteins thus produced localized in the same place in the yeast cell, we could then deduce that Ime4 is present in P-bodies. We observed that Ime4 and Edc3 did not colocalize in the majority of cells, and thus concluded that the Ime4 granules are not P-bodies. However, our experiments showed instances of Ime4 signals near or around the P-bodies in some cells. Hence, the Ime4-containing aggregates are not likely to be P-bodies but could rather represent a different type of granule.

Two distinct nonapeptide systems, consisting of the vasotocin- and oxytocin-related peptides have evolved in vertebrates, and their role in male reproduction is well-described in mammals. In contrast, their comparative role in reproduction in basal vertebrate species, and teleost fishes in particular, has not been investigated in great detail. Using two cyprinid species, the zebrafish (D. rerio) and the goldfish (C. auratus), I address the hypothesis that the teleost nonapeptides vasotocin and isotocin stimulate male cyprinid reproductive physiology by affecting central neuronal and/or peripheral endocrine pathways. To test this hypothesis in zebrafish, an indeterminate breeder, I conducted pharmacological inhibition experiments employing vasotocin and isotocin-specific antagonists in males, a treatment predicted to inhibit reproductive success in mating trials. Because nonapeptides can act both as central peptide neuromodulators and as secreted hormone, I further quantified indices of male courtship behavior (nudging, circling and chasing) and major androgens (testosterone and 11-keto-testosterone) as key endocrine indices of the male reproductive axis. Together, these experiments revealed a dose-dependent, differential inhibition of spawning success, with significant reductions (-65%) in egg fertilization rate observed in pairs in which males had been i.p. injected with 5 ng/g vasotocin and significant reductions (-79%) observed at 500 ng/g i.p injected isotocin. In either case, these partial inhibitions of reproductive success were correlated with significant decreases in specific indices of male courtship behavior, but not endocrine indices, suggesting that individual nonapeptides mediate their effects via central modulation of behavioural neurocircuits. Interestingly, a co-administration of vasotocin and isotocin antagonists completely abolished reproductive success, however this effect was neither correlated with decreases in male courtship behavior, nor endocrine indices, suggesting a separate mode of action, possibly at the level of male pheromone release. To further probe the role of nonapeptides in male zebrafish reproduction, I subsequently tested the hypothesis that nonapeptide systems are acutely activated by key reproductive cues, specifically the releaser pheromone PGF2α, which serves as a chemoattractant and acutely stimulates male reproductive behavior in male cyprinids. Using a chemoattractant choice assay in conjunction with immunohistochemistry and gene expression approaches, I determined whether male zebrafish are attracted to pheromonal cues and acutely activate isotocinergic neurons in the short term and/or regulate nonapeptide gene expression in the longer term. My results show that individual male zebrafish are attracted to PGF2α in an acute choice test. Furthermore, an increase in p-ERK immunoreactivity, a marker of neuronal activation, was observed in the olfactory bulb 10 min following exposure, suggesting a specific response to the pheromone compared to EtOH vehicle. However, no co-localization of p-ERK and IT-positive perikarya was observed in the preoptic area (POA), refuting the hypothesis that PGF2α exposure acutely activates isotocinergic neurons in zebrafish. Analysis of whole brain relative mRNA transcript abundance revealed that PGF2α exposure time-dependently regulates whole brain isotocin, but not vasotocin transcript abundance, suggesting secondary longer-term effects of PGF2α exposure on the isotocinergic system. Using an analogous experimental approach, I further tested the hypothesis that nonapeptides stimulate male reproductive physiology in goldfish, a determinate breeder. Sexually mature male goldfish pretreated with saline or vasotocin or isotocin antagonists were exposed to saline or PGF2α-injected stimulus females and male courtship behavior (chasing, circling), endocrine indices (circulating testosterone) and milt release were quantified. Both nonapeptide antagonists reduced strippable male milt quantity in response to PGF2α-injected females, suggesting a neuronal or hormonal action of both nonapeptides on goldfish milt release. Together, I show that nonapeptides contribute to male reproductive physiology in two species of cyprinids with different reproductive tactics. However, the mode of action may differ from one species to another, with evidence suggesting that nonapeptides play a role in the regulation of reproductive behavior and, possibly, male pheromone, release in zebrafish, while effects on male goldfish seem to be exclusively related to the release of milt. Future studies should compare other teleost species with specific reproductive biology and focus on the gonadal roles of nonapeptides in sperm maturation and/or release.

Two field experiments determined an effective intertrap distance (ITD) for early detection and delimitation sparse gypsy moth (Lepidoptera: Lymantriidae, Lymantria dispar L.) populations in mountainous terrain. This study found that current Animal and Plant Health Inspection Service trapping guidelines are not sufficient for early detection of small gypsy moth populations in mountainous terrain. Detection trapping in mountainous terrain should have an ITD of not more than 804 m. Delimiting trapping should use a grid design with an ITD of 152 m. A related study determined natural adult male mortality in the climate of the intermountain West, which includes Utah, Nevada, western Wyoming, and southern Idaho. An interaction was found between mortality, temperature, and humidity. During high temperatures, most mortality occurred on the second day. When lower temperatures prevailed, the largest percent mortality occurred on the third or fourth day.

This dissertation examines some important physiological and pheromonal factors regulating foraging and colony growth in honey bee colonies. The first study analyzed effects of soybean trypsin inhibitor (SBTI) on the development of hypopharyngeal gland, midgut enzyme activity and survival of the honey bee. In this study newly emerged caged bees were fed pollen diets containing three different concentrations of SBTI. Bees fed 1% SBTI had significantly reduced hypopharyngeal gland protein content. This study indicated that nurse bees fed a pollen diet containing at least 1% SBTI would be poor producers of larval food. In the second study nurse bee biosynthesis of brood food was manipulated using SBTI, and the resulting effects on pollen foraging were measured. Experimental colonies were given equal amounts of SBTI treated and untreated pollen. SBTI treatments had significantly lower hypopharyngeal gland protein content than controls. There was no significant difference in the ratio of pollen to non-pollen foragers and pollen load weights collected between the treatments. These results supported the pollen foraging effort predictions generated from the direct independent effects hypothesis. In the third study we tested whether brood pheromone (BP) regulated queen egg laying via modulation of worker-queen interactions and nurse bee rearing behaviors. This experiment had BP and control treatments. Queens in the BP treatment laid greater number of eggs, were fed for a greater amount of time and were less idle. Significantly more time was spent in cell cleaning by the bees in BP treatments. The results suggest that brood pheromone regulated queen egg-laying rate by modulating worker-queen interactions and nurse bee rearing behavior. The final study of this dissertation focused on how dose-dependent BP-mediated division of labor affected the partitioning of non-foraging and foraging work forces and the amount of brood reared. Triple cohort colonies were used and there were three treatments, Low BP, High BP and Control. Low BP treatments had significantly higher ratio of pollen to non-pollen foragers and greater pollen load weights. Low BP treatment bees foraged at a significantly younger age. This study has shown that BP elicits dose-dependent modulation of foraging and brood rearing behaviors.

This thesis reports the effects of disturbance and harbourage on the fitness of Tribolium confusum and T. castaneum, as well as the the efficacy of pheromone monitoring traps for monitoring for populations of Tribolium in laboratory and mill settings. Behavioural studies were also carried out on mill and laboratory-reared beetles and the distributions of both species in a mill were examined. Twenty-four hour sieving disturbance decreased the rates of dispersal for both species, and decreased T. castaneum fecundity. Rolling disturbance decreased T. confusum dispersal rate while shaking disturbance decreased T. castaneum dispersal rate. When undisturbed beetles were given differing amounts of flour in the presence or absence of harbourage, beetles laid more eggs in larger amounts of flour, but harbourage only affected T. castaneum at one level of flour (2 g). Throughout disturbance and harbourage experiments, T. castaneum laid more eggs than T. confusum. Pheromone monitoring traps placed in three Canadian flour mills were not useful in predicting the degree of infestation inside Simons rollstands. Pheromone monitoring traps also showed low efficacy (i.e. caught few beetles) in both mill and laboratory settings, and T. confusum was caught less often than T. castaneum in both mills and in a warehouse. Mill-strain beetles of both species were caught less often than laboratory-strain beetles in a warehouse. In one Canadian flour mill, both T. castaneum and T. confusum were found inside rollstands but the two species were spatially segregated from one another, rarely being found together in the same rollstand. In contrast to this, both species were consistently found together in samples taken from the same mill less than a year beforehand. In behavioural laboratory studies, beetles collected directly from a mill moved slower than beetles collected from a laboratory culture and this response was shown to be phenotypic. Mill-strain and laboratory-strain beetles also differed in burrowing tendencies, with T. confusum from the laboratory strain burrowing less than T. confusum from a mill and T. castaneum from different mills sometimes burrowing more and sometimes less than T. castaneum from the laboratory strain. / October 2008

This thesis presents a novel algorithm called ant colony optimization with dual pheromone tables (ACODPT) for improving the quality of ant colony optimization (ACO). The proposed algorithm works by adding a so-called ¡§negative¡¨ pheromone table to ACO to avoid the problem of ACO easily falling into local optima. By using the ¡§negative¡¨ pheromone table to eliminate the most impossible path to search for the new solution, the probability of selecting the remaining paths is increased, and so is the quality. To evaluate the performance of the proposed algorithm, ACODPT is compared with several state-of-the-art algorithms in solving the clustering problem. The experimental results show that the proposed algorithm can eventually prevent ACO from falling into local optima in the early iterations, thus providing a better result than the other algorithms in many cases.

This dissertation examines some important physiological and pheromonal factors regulating foraging and colony growth in honey bee colonies. The first study analyzed effects of soybean trypsin inhibitor (SBTI) on the development of hypopharyngeal gland, midgut enzyme activity and survival of the honey bee. In this study newly emerged caged bees were fed pollen diets containing three different concentrations of SBTI. Bees fed 1% SBTI had significantly reduced hypopharyngeal gland protein content. This study indicated that nurse bees fed a pollen diet containing at least 1% SBTI would be poor producers of larval food. In the second study nurse bee biosynthesis of brood food was manipulated using SBTI, and the resulting effects on pollen foraging were measured. Experimental colonies were given equal amounts of SBTI treated and untreated pollen. SBTI treatments had significantly lower hypopharyngeal gland protein content than controls. There was no significant difference in the ratio of pollen to non-pollen foragers and pollen load weights collected between the treatments. These results supported the pollen foraging effort predictions generated from the direct independent effects hypothesis. In the third study we tested whether brood pheromone (BP) regulated queen egg laying via modulation of worker-queen interactions and nurse bee rearing behaviors. This experiment had BP and control treatments. Queens in the BP treatment laid greater number of eggs, were fed for a greater amount of time and were less idle. Significantly more time was spent in cell cleaning by the bees in BP treatments. The results suggest that brood pheromone regulated queen egg-laying rate by modulating worker-queen interactions and nurse bee rearing behavior. The final study of this dissertation focused on how dose-dependent BP-mediated division of labor affected the partitioning of non-foraging and foraging work forces and the amount of brood reared. Triple cohort colonies were used and there were three treatments, Low BP, High BP and Control. Low BP treatments had significantly higher ratio of pollen to non-pollen foragers and greater pollen load weights. Low BP treatment bees foraged at a significantly younger age. This study has shown that BP elicits dose-dependent modulation of foraging and brood rearing behaviors.

Brood pheromone of the honey bee, (Apis mellifera) has been shown to elicit a wide array of primer and releaser effects on non‐foragers and foragers leading to the regulation of nursing, pollen foraging, and behavioral development such that the behavior of the colony may be regulated by the amount and condition of the larvae. To date, all studies into the effects of brood pheromone have either used uncharacterized whole extracts or a single blend of brood pheromone characterized from a population of honey bees in France. The variation in the relative proportions of the ten fatty‐acid ester components that characterize brood pheromone and some effects of this variation on pollen foraging and sucrose response thresholds were therefore observed. The objectives met in this dissertation were to determine whether changes in brood pheromone component proportions (blend) or amount communicates larval nutritional status and reports the results of observations of nurses and foragers in response to blends of brood pheromone from deprived and‐non deprived larvae, to measure how changes in brood pheromone blend changed pollen foraging behavior and if such changes could account for the pollen foraging differences between Africanized and European bees, and finally to observe the effects of exposure time on brood pheromone blend and to observe whether non‐foragers made contact with the pheromone. Brood pheromone was found to vary by larval rearing environment, but did not elicit the expected behaviors that would support a cue of nutritional status. Brood pheromone also varied significantly by mitochondrial lineage/population source and responses to brood pheromone appeared to be coadapted to blend, suggesting that brood pheromone may be important in race recognition. Finally, brood pheromone varied significantly over time and was found to be removed from sources by bees, suggesting possible mechanisms for loss of effect. Combined the results of this research indicate that brood pheromone blend differences lead to profound changes in colony behavior related to pollen foraging and food provisioning, providing novel tools for colony manipulation and mechanisms for understanding brood rearing division of labor and chemical communication.

The aggregation-attachment-pheromone (AAP) of two geographic strains of the Gulf Coast tick, Amblyomma maculatum Koch, was investigated to evaluate practicality of using solid-phase-microextraction (SPME) in an AAP study of Gulf Coast tick. Solid-phase microextraction was used to compare the AAP production in two strains of fed male Gulf Coast tick and demonstrate and confirm the presence of AAP in bioassays. A solid-phase-microextraction (SPME) headspace collection technique was sufficient to capture volatile organic compounds produced by fed and unfed male Gulf Coast ticks. Gas chromatography analysis revealed three major volatile organic compounds were produced in significantly greater amounts (p < 0.05) by fed males than those produced by unfed males. These volatile compounds were produced in significantly higher amount by the third day of feeding by male ticks. However, two of these volatiles remained relatively constant in their production while the primary volatile compound increased in its production until the eighth day of feeding by male Gulf Coast ticks. Also, the relative abundances of these three volatile organic compounds were different between Oklahoma and Texas strains of Gulf Coast ticks. The activity of AAP from fed male Gulf Coast ticks was confirmed using two bioassay techniques. A petri dish bioassay revealed significantly higher numbers of female Gulf Coast ticks attracted to fed-males which also produced significantly greater amounts (p < 0.001) of volatile organic compounds detected by GC analysis. The Ytube olfactometer bioassay revealed that significantly higher numbers of females responded to fed-males or to CO2 when compared to purified air (p < 0.001), but the differences in female response to fed-males and CO2 were not significant (p < 0.391 in Oklahoma strain and p < 0.458 in Texas strain). However, female responses to stimuli containing both fed-males and CO2 were significantly higher when compared to either stimulus alone (p < 0.001).