Ants are ubiquitous in most communities and many form opportunistic mutualisms with honeydew-producing hemipterans (e.g. treehoppers). Hemipterans excrete honeydew, a carbohydrate rich substance, that ants harvest and, in return, ants protect their honeydew-producing partners from parasitoids, predators, and competitors. Given the efficacy of tending ants in removing hemipteran antagonists, and the strong roles that ants play within their communities as predators, competitors, and seed dispersers, surprisingly little is known of the effects of ant-hemipteran mutualisms (AHM) on the invertebrate communities in which they are embedded or on the plants that host AHM. Using observational and manipulative field experiments, I examined the long-term effect of AHM on their host plant’s, honey mesquite (Prosopis glandulosa), reproductive potential and quality. In addition, I measured how the presence of AHM affects the abundance, richness, diversity, and composition of the invertebrate communities living on honey mesquite. Plants hosting AHM may indirectly benefit (through the removal of herbivore arthropods) or suffer (through the loss pollinators) due to the defensive behavior of tending ants. To determine the effects of AHM on their host plant, I established a four-year press experiment in which I removed AHM from 50 randomly trees, while leaving 50 as controls. In addition, I marked and followed 30 trees from which AHM were naturally absent. To assess if mesquite quality differed between trees hosting AHM and trees in which AHM were naturally absent, in 2012 I assayed foliar condensed tannin concentrations, a secondary defense compound, and, in 2015, I measured foliar nitrogen, phosphorous, potassium, magnesium, and iron as they are essential for growth and reproduction. I compared the reproductive potential between AHM present and removed trees by counting flowers and fruits across all 4 years of the study. Mesquite that hosted AHM contained significantly less condensed tannins and significantly higher concentrations of N%, Mg, and Fe. Furthermore, over the duration of the study mesquite hosting AHM contained significantly more flowers than those from which AHM were removed or naturally absent. My results indicate that AHM select trees of high quality and their continued presence is associated with high levels of reproductive potential. Most studies that have evaluated community-level effects of AHM compare total abundance and species richness in communities (or host plants) with and without AHMs. However, both measures are dependent on sampling effort, complicating comparisons across different studies. To examine the effects of AMH on the arthropod community in mesquite, I first compared family richness and alpha diversity using standardized rarefaction and extrapolation curves. I then measured beta diversity and turnover in community composition from one year to the next. The removal of AHM increased invertebrate diversity and significantly altered community composition. Although treatments did not statistically differ in turnover rates, replacements occurred among treatments at the family level which may be biologically meaningful. Furthermore, herbivore and predator populations increased, and pollinator populations decreased following the removal of AHM. These results suggest that the presence of AHM can alter the composition of arthropod communities and food-web dynamics. However, these effects were significant in some years and not others, suggesting the importance of temporal variation in drivers of communities. Overall, my work demonstrates that AHM can be drivers of community composition and illustrate the importance of examining their effects across multiple seasons.

In insects, ecdysis is an important feature of growth and development and is tightly controlled by a variety of neuropeptides. In holometabolous insects, crustacean cardioactive peptide (CCAP) is one of many factors that regulate ecdysis behaviours; however, not much is known about the control of ecdysis in hemimetabolous insects. In this thesis, the CCAP-signaling pathway is shown to be essential for successful ecdysis in the hemimetabolous insect, Rhodnius prolixus. The cDNA sequence of the CCAP gene has been cloned from the R. prolixus central nervous system (CNS) and the functional role of CCAP as a neuromodulator/neurotransmitter demonstrated. Specifically, the expression of RhoprCCAP in CNS neurons producing extensive CCAP-like immunoreactive processes within the neuropile indicates that CCAP plays central roles in coordination of other neurons. RhoprCCAP also acts as a neurohomone/neuromodulator released peripherally to coordinate many tissues. Thus, CCAP-like immunoreactive processes are found in neurohemal sites and also on peripheral tissues. The RhoprCCAP receptor (RhoprCCAPR) has been cloned and shown to be a G-protein coupled receptor (GPCR). RhoprCCAPR expression is observed in the CNS and certain peripheral tissues of R. prolixus. Also, CCAP stimulates hindgut contractions and increases the heartbeat rate in a dose-dependent manner. The involvement of CCAP in R. prolixus ecdysis has been investigated. Up-regulation of the RhoprCCAP transcript in the CNS and the RhoprCCAP receptor (RhoprCCAPR) transcript in the CNS and specific peripheral tissues was observed immediately prior to ecdysis. Also, decreasing staining intensity of CCAP-like immunoreactivity in neurons immediately following ecdysis indicates the release of CCAP during ecdysis. The critical importance of the CCAP-signalling pathway was further demonstrated by knockdown of the RhoprCCAP and RhoprCCAPR transcripts utilizing double stranded RNA interference. Insects with these transcripts knocked down have high mortality (up to 84%), typically at the expected time of ecdysis, or have ecdysis extremely delayed. Taken together, this thesis demonstrates that RhoprCCAP plays a crucial role in regulating ecdysis behaviours in R. prolixus, and clearly shows the conserved nature of the CCAP-signaling pathway in ecdysis for both holometabolous and hemimetabolous insects.

In insects, ecdysis is an important feature of growth and development and is tightly controlled by a variety of neuropeptides. In holometabolous insects, crustacean cardioactive peptide (CCAP) is one of many factors that regulate ecdysis behaviours; however, not much is known about the control of ecdysis in hemimetabolous insects. In this thesis, the CCAP-signaling pathway is shown to be essential for successful ecdysis in the hemimetabolous insect, Rhodnius prolixus. The cDNA sequence of the CCAP gene has been cloned from the R. prolixus central nervous system (CNS) and the functional role of CCAP as a neuromodulator/neurotransmitter demonstrated. Specifically, the expression of RhoprCCAP in CNS neurons producing extensive CCAP-like immunoreactive processes within the neuropile indicates that CCAP plays central roles in coordination of other neurons. RhoprCCAP also acts as a neurohomone/neuromodulator released peripherally to coordinate many tissues. Thus, CCAP-like immunoreactive processes are found in neurohemal sites and also on peripheral tissues. The RhoprCCAP receptor (RhoprCCAPR) has been cloned and shown to be a G-protein coupled receptor (GPCR). RhoprCCAPR expression is observed in the CNS and certain peripheral tissues of R. prolixus. Also, CCAP stimulates hindgut contractions and increases the heartbeat rate in a dose-dependent manner. The involvement of CCAP in R. prolixus ecdysis has been investigated. Up-regulation of the RhoprCCAP transcript in the CNS and the RhoprCCAP receptor (RhoprCCAPR) transcript in the CNS and specific peripheral tissues was observed immediately prior to ecdysis. Also, decreasing staining intensity of CCAP-like immunoreactivity in neurons immediately following ecdysis indicates the release of CCAP during ecdysis. The critical importance of the CCAP-signalling pathway was further demonstrated by knockdown of the RhoprCCAP and RhoprCCAPR transcripts utilizing double stranded RNA interference. Insects with these transcripts knocked down have high mortality (up to 84%), typically at the expected time of ecdysis, or have ecdysis extremely delayed. Taken together, this thesis demonstrates that RhoprCCAP plays a crucial role in regulating ecdysis behaviours in R. prolixus, and clearly shows the conserved nature of the CCAP-signaling pathway in ecdysis for both holometabolous and hemimetabolous insects.

Rhodnius prolixus, the principal Chagas disease vector, requires a blood meal to complete its moult cycle into the next stage. Allatotropins (ATs), a family of peptides first isolated from Manduca sexta, have been shown to regulate the biosynthesis of juvenile hormone, an insect growth and development hormone; however, ATs, being multimodal peptides, also exhibit myotropic effects on some insect visceral muscles. Here, this AT family of peptides has been examined in R. prolixus. Genomic analysis revealed a cDNA fragment of 973bp encoding one mature amidated AT tridecapeptide (Rhopr-AT) with high transcript levels observed, via RT-PCR, in the central nervous system (CNS) and pool of fat body and trachea. AT-like immunoreactive neurons were found throughout the CNS and AT-like immunoreactive processes were present on some peripheral tissues. Bioassays using hindgut and dorsal vessel contraction, however, failed to demonstrate any effect of Rhopr-AT on these tissues. Future work will examine the effects of Rhopr-AT on JH production.

Rhodnius prolixus, the principal Chagas disease vector, requires a blood meal to complete its moult cycle into the next stage. Allatotropins (ATs), a family of peptides first isolated from Manduca sexta, have been shown to regulate the biosynthesis of juvenile hormone, an insect growth and development hormone; however, ATs, being multimodal peptides, also exhibit myotropic effects on some insect visceral muscles. Here, this AT family of peptides has been examined in R. prolixus. Genomic analysis revealed a cDNA fragment of 973bp encoding one mature amidated AT tridecapeptide (Rhopr-AT) with high transcript levels observed, via RT-PCR, in the central nervous system (CNS) and pool of fat body and trachea. AT-like immunoreactive neurons were found throughout the CNS and AT-like immunoreactive processes were present on some peripheral tissues. Bioassays using hindgut and dorsal vessel contraction, however, failed to demonstrate any effect of Rhopr-AT on these tissues. Future work will examine the effects of Rhopr-AT on JH production.

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