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Description
The 18S ribosomal RNA gene is ubiquitous across eukaryotes as it encodes the RNA component of the ribosomal small subunit. It is the most commonly used marker in molecular studies of unicellular eukaryotes (protists) due to its species specificity and high copy number in the protist genome. Recent studies have revealed the widespread occurrence of intragenomic (intra-individual) polymorphism in many protists, an understudied phenomenon which contradicts the assumed homogeneity of the 18S throughout an individual genome. This thesis quantifies and analyzes the level of intragenomic and intraspecific 18S sequence variability in three Trichonympha species (T. campanula, T. collaris, T. postcylindrica) from Zootermopsis termites. Single-cell DNA extractions, PCR, cloning, and sequencing were performed to obtain 18S rRNA sequence reads, which were then analyzed to determine levels of sequence divergence among individuals and among species. Intragenomic variability was encountered in all three species. However, excluding singleton mutations, sequence divergence was less than 1% in 53 of the 56 compared individuals. T. collaris exhibited the most substantial intragenomic variability, with sequence divergence ranging from 0 to 3.4%. Further studies with more clones per cell are needed to elucidate the true extent of intragenomic variability in Trichonympha.
ContributorsBobbett, Bradley (Author) / Gile, Gillian (Thesis director) / Liebig, Juergen (Committee member) / School of International Letters and Cultures (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Adaptation of Camponotus floridanus’ Cuticular Hydrocarbon Profile under High Temperature Conditions
Description
Insects are small creatures highly susceptible to water loss. A major factor in the prevention of water loss through an insect’s cuticle are their cuticular hydrocarbons (CHC), a lipid layer consisting mostly of long-chain hydrocarbons. CHCs consist of different molecules called alkanes, alkenes, and methyl branched hydrocarbons which all have varying levels of hydrophobicity. Ants are a massively abundant family of insects with important roles in the ecosystem that also utilize CHCs. Camponotus floridanus isare athe native ant species of the Florida Keys which areis known to have variable environmental temperature. Being exposed to temperatures as high as 35 °C, these ants are expected to have mechanisms that allow them to adapt to their environment. It was hypothesized that CHCs may change in concentration or composition as a means to combat the changes in cuticular permeability due to the variable temperatures that the ants experience. We therefore used C. floridanus worker ants to learn more about CHC plasticity in insects when exposed to elevated temperatures. We found four CHC componentspeaks that showed a statistically significant increase in concentration when comparing the control to treatment colonies: 3,7 dimethyl C31, an underdetermined methyl branched C31, 3,7,11 trimethyl C31, and an undetermined tetramethylbranched C31. These significant changes in concentration occurred on longer chain hydrocarbons. Under further examination, it was found that there was a strong positive correlation between elution time and the differences in medians of peak area between control and treatment colonies. This shows that there was a shift in the CHC profile resulting in an increased concentration of longer chained methyl-branched hydrocarbons. It also suggests that branched hydrocarbons also play some role in the water proofing mechanism of C. floridanus.
ContributorsOn, Thomas (Co-author) / On, Tyler (Co-author) / Liebig, Juergen (Thesis director) / Harrison, Jon (Committee member) / Murdock, Tyler (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
When ants encounter liquid food, they have two options of transporting that food to their nests. The first is the social bucket method in which liquid is carried in the mandibles of the workers back to the nest. The second is trophallaxis in which liquid is imbibed and then transported inside the ant back to the nest. The liquid is then regurgitated to fellow nestmates. Ectatomma have been observed using the social bucket method of transport and were considered members of the Ponerine family. However, a new phylogeny created by Borowiec and Rabeling places Ectatomma near to Formecinae and Myrmicinae, both know for practicing trophallaxis. This seems to suggest either Ectatomma is able to utilize trophallaxis as well or that the evolutionary practice of trophallaxis is more plastic than previously believed. The ability of Ectatomma ruidum to utilize trophallaxis was examined in two experiments. The first experiment examined E. ruidum’s ability to practice worker to worker trophallaxis and the second examined E. ruidum’s ability to perform worker to larva trophallaxis. The results of both experiments indicated that E. ruidum cannot utilize trophallaxis but the larva of E. ruidum may be able to regurgitate to the workers. These results in turn seem to suggest that trophallaxis is a bit more plastic than originally thought.
ContributorsCunningham, Cassius Alexander (Author) / Pratt, Stephen (Thesis director) / Liebig, Juergen (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Spaceflight and spaceflight analogue culture enhance the virulence and pathogenesis-related stress resistance of the foodborne pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium). This is an alarming finding as it suggests that astronauts may have an increased risk of infection during spaceflight. This risk is further exacerbated as multiple studies indicate that spaceflight negatively impacts aspects of the immune system. In order to ensure astronaut safety during long term missions, it is important to study the phenotypic effects of the microgravity environment on a range of medically important microbial pathogens that might be encountered by the crew. This ground-based study uses the NASA-engineered Rotating Wall Vessel (RWV) bioreactor as a spaceflight analogue culture system to grow bacteria under low fluid shear forces relative to those encountered in microgravity, and interestingly, in the intestinal tract during infection. The culture environment in the RWV is commonly referred to as low shear modeled microgravity (LSMMG). In this study, we characterized the stationary phase stress response of the enteric pathogen, Salmonella enterica serovar Enteritidis (S. Enteritidis), to LSMMG culture. We showed that LSMMG enhanced the resistance of stationary phase cultures of S. Enteritidis to acid and thermal stressors, which differed from the LSSMG stationary phase response of the closely related pathovar, S. Typhimurium. Interestingly, LSMMG increased the ability of both S. Enteritidis and S. Typhimurium to adhere to, invade into, and survive within an in vitro 3-D intestinal co-culture model containing immune cells. Our results indicate that LSMMG regulates pathogenesis-related characteristics of S. Enteritidis in ways that may present an increased health risk to astronauts during spaceflight missions.
ContributorsKoroli, Sara (Author) / Nickerson, Cheryl (Thesis director) / Barrila, Jennifer (Committee member) / Ott, C. Mark (Committee member) / School of Life Sciences (Contributor) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
An intimate view of the unique architecture of Harpegnathos saltwater nest using aluminum nest casts
Description
Abstract:
Given the incredible variety in ant nest architecture, this experiment sought to evaluate how the nest architecture of Harpegnathos saltator differs from other species’ nests. To achieve the ability to evaluate the structure of H. saltator nest, we created experimental colonies varying in size from 20, 40, 60, 80 workers of Harpegnathos saltator in five-gallon buckets of sand and then allowing the colonies to grow for four months and twelve days. To create the nest casts, we developed a charcoal kiln out of a galvanized trash can and used a ceramic crucible to hold the aluminum being melted. Using molten aluminum to create nest casts of each colony produced, we obtained three poorly developed nests and one decent nest. The decent nest cast, the 80 worker H. saltator nest, was lacking key features of H. saltator nests that have been excavated in the field. However, they do share many of the same structures such as the shaping of the chambers. The ability of the experimental colonies to excavate the soil provided in the buckets to them was likely halted by poor penetration of water into superficial layers of the soil, thus making the soil too difficult to excavate and form the structures that are key elements of the species nest architecture. Despite these key challenges which the colonies faced, the 80-worker colony showed extensive vertical development and did display features associated with natural H. saltator colonies. Thus, given the display of some key features associated with characteristics of the H. saltator nests excavated in the field, it can be said that with some modification to technique that this is a viable avenue for future study of nest architecture and colony structure.
Given the incredible variety in ant nest architecture, this experiment sought to evaluate how the nest architecture of Harpegnathos saltator differs from other species’ nests. To achieve the ability to evaluate the structure of H. saltator nest, we created experimental colonies varying in size from 20, 40, 60, 80 workers of Harpegnathos saltator in five-gallon buckets of sand and then allowing the colonies to grow for four months and twelve days. To create the nest casts, we developed a charcoal kiln out of a galvanized trash can and used a ceramic crucible to hold the aluminum being melted. Using molten aluminum to create nest casts of each colony produced, we obtained three poorly developed nests and one decent nest. The decent nest cast, the 80 worker H. saltator nest, was lacking key features of H. saltator nests that have been excavated in the field. However, they do share many of the same structures such as the shaping of the chambers. The ability of the experimental colonies to excavate the soil provided in the buckets to them was likely halted by poor penetration of water into superficial layers of the soil, thus making the soil too difficult to excavate and form the structures that are key elements of the species nest architecture. Despite these key challenges which the colonies faced, the 80-worker colony showed extensive vertical development and did display features associated with natural H. saltator colonies. Thus, given the display of some key features associated with characteristics of the H. saltator nests excavated in the field, it can be said that with some modification to technique that this is a viable avenue for future study of nest architecture and colony structure.
ContributorsAnderson, Clayton Edward (Author) / Liebig, Juergen (Thesis director) / Pratt, Stephen (Committee member) / School of Politics and Global Studies (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Complex animal societies consist of a plethora of interactions between members. To successfully thrive they must be able to recognize members and their kin, and to understand how they do this we need sufficient and reliable methods of testing. Eusocial insects are especially good at recognizing their nestmates, but the exact mechanism or how well they can discriminate is unknown. Ants achieve nestmate recognition by identifying varying proportions of cuticular hydrocarbons. Previous studies have shown ants can be trained to discriminate between pairs of hydrocarbons. This study aims to compare two methodologies previously shown to demonstrate odor learning to identify which one is the most promising to use for future odor learning experiments. The two methods tested were adapted from Sharma et al. (2015) and Guerrieri and d’Ettorre (2010). The results showed that the Guerrieri method demonstrated learning better and was more reliable and faster than the Sharma method. The Guerrieri method should be used in future experiments regarding odor learning discrimination
ContributorsDavis, Cole (Author) / Liebig, Juergen (Thesis director) / Stephen, Pratt (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
For colonies of ponerine ant species, sterility regulation after a founding queen's death is not totally achieved in the worker caste, and the possibility of sexual reproduction is opened to workers. The persisting survival of these colonies is dependent on capturing the optimal reproductive ratio; yet, an informational gap bounds the mechanisms detailing the selection of new reproductives and the suppression of ovarian development in rejected reproductives. We investigated the mechanisms of worker policing, one of the primary methods of ovarian suppression, through continuous video observation for a period of five days at the start of colony instability. Observations suggest policing in H. saltator is performed by a majority of a colony, including potential reproductives, and requires multiple events to fully discourage ovarian growth.
ContributorsChien, Jeffrey (Co-author) / Barat Ali, Fatima (Co-author) / Kang, Yun (Thesis director) / Liebig, Juergen (Committee member) / School of Mathematical and Statistical Sciences (Contributor) / Mechanical and Aerospace Engineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2018-12
Description
Due to the widely accepted trend of urbanization displacing wildlife from their natural habitats and niches, many wildlife conservation organizations have sprouted up, even in Phoenix. Liberty Wildlife Foundation is one that rehabilitates avian wildlife. Several studies have mentioned an opposing theory: that urbanization helps conserve those species that have turned urban environments into a niche of their own. Since these wildlife conservation centers are localized in cities themselves, this brings into question these organizations' definitions of the term "wildlife." This study examined injury and recovery statistics to determine just how many of the patients admitted were conventional wildlife versus urban-dwelling city birds, and whether this classification had any effect on their likeliness of recovery and release. The data showed that out of over 130 species, a few key urban species contributed to an overwhelmingly large majority of injured birds admitted to the center in 2017; urban and non-urban birds, however, had relatively equal average release frequencies, demonstrating then that their likelihood of recovery was predominantly dependent on the injury borne by them.
ContributorsVirdee, Rishika Kaur (Author) / Liebig, Juergen (Thesis director) / Lynch, John (Committee member) / Haight, Kevin (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
The gastrointestinal (GI) tract is home to a complex and diverse microbial ecosystem that contributes to health or disease in many aspects. While bacterial species are the majority in the GI tract, their cohabitants, fungal species, should not be forgotten. Children with autism spectrum disorder (ASD) often suffer from GI disorders and associated symptoms, implying a role the bacterial and fungal gut microbiota play in maintaining human health. The irregularities in GI symptoms can negatively affect the overall quality of life or even worsen behavioral symptoms the children present. Even with the increase in the availability of next-generation sequencing technologies, the composition and diversities of fungal microbiotas are understudied, especially in the context of ASD. We therefore aimed to investigate the gut mycobiota of 36 neurotypical children and 38 children with ASD. We obtained stool samples from all participants, as well as autism severity and GI symptom scores to help us understand the effect the mycobiome has on these symptoms. By targeting the fungal internal transcribed spacer (ITS) and bacterial 16S rRNA V4 regions, we obtained fungal and bacterial amplicon sequences, from which we investigated the diversities, composition, and potential link between two different ecological clades. From fungal amplicon sequencing results, we observed a significant decrease in the observed fungal OTUs in children with ASD, implying a lack of potentially beneficial fungi in ASD subjects. We performed Bray-Curtis principal coordinates analysis and observed significant differences in fungal microbiota composition between the two groups. Taxonomic analysis showed higher relative abundances of Candida , Pichia, Penicillium , and Exophiala in ASD subjects, yet due to a large dispersion of data, the differences were not statistically significant. Interestingly, we observed a bimodal distribution of Candida abundances within children with ASD. Candida's relative abundance was not significantly correlated with GI scores, but children with high Candida relative abundances presented significantly higher Autism Treatment Evaluation Checklist (ATEC) scores, suggesting a role of Candida on ASD behavioral symptoms. Regarding the bacterial gut microbiota, we found marginally lower observed OTUs and significantly lower relative abundance of Prevotella in the ASD group, which was consistent with previous studies. Taken together, we demonstrated that autism is closely linked with a distinct gut mycobiota, characterized by a loss of fungal and bacterial diversity and an altered fungal and bacterial composition.
ContributorsPatel, Jigar (Author) / Krajmalnik-Brown, Rosa (Thesis director) / Kang, Dae Wook (Committee member) / Adams, James (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
The effect of an anaerobic reductive environment produced by the oxidation of zero valent iron (ZVI) on the microbial reductive dechlorination of trichloroethylene and its applicability to in-situ bioremediation processes was investigated using microcosms and soil column studies. I learned that microbial dechlorination requires a highly reductive environment, as represented by negative values for oxidation-reduction potential (ORP), which can be maintained through the addition of reducing agents such as ZVI, or to a lesser extent, the fermentation of added substrates such as lactate. Microcosm conditions represented distance from an in-situ treatment injection well and contained different types of iron species and dechlorinating bioaugmentation cultures. Diminishing efficacy of microbial reductive dechlorination along a gradient away from the injection zone was observed, characterized by increasing ORP and decreasing pH. Results also suggested that the use of particular biostimulation substrates is key to prioritizing the dechlorination reaction against competing microbial and abiotic processes by supplying electrons needed for microbial dechlorination.
ContributorsMouti, Aatikah (Author) / Krajmalnik-Brown, Rosa (Thesis director) / Delgado, Anca (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2017-12