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Item Microbiomes and zoonoses: dynamics of the black flying fox (Pteropus alecto) gastrointestinal microbiome(Montana State University - Bozeman, College of Agriculture, 2022) Jones, Devin Nicole; Chairperson, Graduate Committee: Raina K. Plowright; This is a manuscript style paper that includes co-authored chapters.Land-use change is increasingly recognized as a driver of spillover of zoonotic pathogens. Australian black flying foxes (Pteropus alecto) are experiencing extensive loss of habitat which reduces available food, particularly in winter. Hendra virus (HeV, family: Paramyxoviridae) was isolated from horses and humans in 1994 and P. alecto was later identified as the reservoir host. As habitat loss threatens these bat populations, and Hendra virus continues to spill over to horses annually, it is important to understand factors that influence bat health and viral shedding. Because gastrointestinal tract (GIT) microbiomes are important for host health and are understudied in flying foxes, the goal of this research was to understand the natural dynamics of the P. alecto GIT microbiome and its associations with diet, body composition, markers of inflammation, and viral shedding. We sampled Pteropus alecto near Brisbane from 2018-2020. We captured bats returning from foraging and collected rectal swabs to determine the GIT microbiome using 16S rRNA amplicon sequencing. In addition to feces for dietary analysis, we also collected samples to measure health and infection, including blood to measure neutrophil-to-lymphocyte ratios, urine to detect Hendra virus, and bioelectrical impedance analysis to measure body fat. These data enabled us to determine how the P. alecto GIT microbiome varied within individuals over time and in the context of physiological, ecological, and dietary shifts. Lastly, we asked if we could predict health outcomes using the GIT microbiome. We found that P. alecto GIT microbiomes are highly dynamic over time, through different life stages, between foraging strategies, and that the type of diet is associated with GIT microbiome diversity. Bats consuming native foods had lower GIT microbiome diversity compared to those consuming introduced and cultivated foods. Despite associations between body fat and HeV infection, the GIT microbiome was not able to predict these health outcomes. These results suggest that P. alecto GIT microbiomes are highly dynamic and may not contribute significantly to host health. Future research should incorporate more health metrics or other approaches to microbiome profiling to determine if the GIT microbiome could be used as a biomarker of health.Item Using gastrointestestinal organoids to study infectious diseases in humans and bats(Montana State University - Bozeman, College of Agriculture, 2021) Hashimi, Marziah; Chairperson, Graduate Committee: Diane Bimczok; This is a manuscript style paper that includes co-authored chapters.The gastrointestinal epithelium plays a critical role in protecting the gastrointestinal mucosa from invading microorganism such as bacteria or a viruses. Helicobacter pylori (H. pylori) infection of human gastric epithelium causes gastric cancer, which is the third leading cause of cancer-related mortality worldwide. Dendritic cells (DCs)--which are antigen presenting cells--are responsible for the activation of T cells. However, the mechanism by which DCs are recruited to the gastric epithelium is still unknown. We hypothesized that the DCs are recruited to the gastric epithelium in a chemokine- dependent manner. For my thesis work, I utilized human primary gastric epithelial organoids cells to test this hypothesis and evaluate the recruitment of DCs to the epithelium under normal conditions and upon H. pylori infection. Using monocyte-derived DCs in a chemotaxis assay, I showed that these cells are recruited to H. pylori-infected organoid supernatant. I showed that this recruitment is chemokine- dependent, as it was significantly decreased when a chemokine receptor inhibitor was included in the chemotaxis assay. COVID-19 is caused by severe respiratory syndrome coronavirus-2 (SARS-CoV-2). In addition to respiratory symptoms, COVID-19 patients can also have diarrhea and vomiting, indicating that SARS-CoV-2 may infect the gastrointestinal tract. Bats are thought to be the natural reservoirs for SARS-CoV-2, however there is no known bat gastrointestinal model to study SARS-CoV-2 infection. In the second part of my thesis, I developed Jamaican fruit bat (JFB), Artibeus jamaicensis) gastrointestinal organoids (JFB organoids). I successfully developed organoids from JFB stomach, proximal and distal intestine. I showed via histology and gene expression that developed organoids do indeed recapitulate their corresponding tissues from which they were derived. I also tested whether the JFB distal intestinal organoids were susceptible to SARS-CoV-2 infection. While they do not support the active replication of SARS-CoV-2 infection, they did show antiviral and pro-inflammatory responses. My results also showed that SARS-CoV-2 does not induce programmed cell death in the organoids.Item Effect of landscape fragmentation on bat population dynamics and disease persistence in Uruguay(Montana State University - Bozeman, College of Agriculture, 2021) Botto Nunez, German; Chairperson, Graduate Committee: Raina K. Plowright; This is a manuscript style paper that includes co-authored chapters.The transmission of pathogens into novel host species, a process known as spillover, requires a series of conditions to align in space and time. A series of imperfect barriers prevent the jump of pathogens from one species to others. These may include the distribution and abundance of the primary host, the survival of the pathogen in the environment and the susceptibility of the recipient host to the pathogen. Only when permissive conditions align in time and space can the spillover occur. Spillovers may be relatively rare events and the understanding of the dynamics of the barriers is constrained by the ability of detecting and analyzing such events. Systems where spillover does not occur, despite apparent presence of all required conditions, provide an opportunity to understand barriers preventing inter-species transmission. Vampire bat-borne rabies in Uruguay provides such an opportunity. Despite large and stable livestock density, presence of vampire bats, and circulation of the virus in close proximity, the country did not experience livestock rabies outbreaks until 2007. Here we combined historical review, field sampling, and statistical and mathematical modeling to understand the factors driving the emergence of rabies in Uruguay in 2007 and the previous absence of the disease. Our results suggest that rabies outbreaks in the country are spatially and temporally associated with fragmentation of grasslands. We showed that proposed increased connectivity among colonies, in response to fragmentation, is sufficient to explain longer persistence of the virus in the bat colonies, allowing more opportunities for virus transmission to livestock. We showed that connectivity has a strong effect on rabies persistence and that reproductive seasonality and population turnover have only marginal effects compared to connectivity. As connectivity driven by shared feeding areas might not be detectable by genetic analyses of the bats, we proposed the use of a widespread virus persistently infecting bats as a marker to trace connectivity across colonies. Combined, the results presented here provide tools that can be applied to intervene and apply countermeasures to prevent spillover.Item Characterizing summer roosts of male little brown myotis (Myotis lucifugus) in lodgepole pine-dominated forests(Montana State University - Bozeman, College of Letters & Science, 2020) Hilty, Shannon Lauree; Chairperson, Graduate Committee: Andrea LittAlthough bat roosts have been well-studied in the eastern United States, we know less about roosts in the west. Western bats may make use of trees and snags, as in the eastern US, but the Rocky Mountains provide more exposed rock, which could contribute to western bat species using different roosting features. Additionally, roost studies often focus on maternity colonies, and information on roosts used by male bats is limited. Given that roosting sites may be limiting, we aimed to quantify structural features of roosts used by male little brown myotis (Myotis lucifugus) in forests dominated by lodgepole pine (Pinus contorta) during the summer and determine whether bats are selecting roosts with particular features disproportionately to what is available on the landscape. We mist-netted for bats during the summers of 2017 and 2018 and attached transmitters to 34 male little brown myotis. We located at least 1 roost for 20 individuals (average = 2.85 roosts/bat, range = 1-6). Although snags were available, most bats roosted in rock features (86% in rocks, 14% in snags); rock roosts were mainly in crevices with vertical orientations (85%) instead of rock cavities (15%). Male bats were more likely to select roosts with less canopy closure (mean for used locations = 14.1%, SE = 2.3) that were closer to water (1063.1 m, SE = 136.2). They also selected roosts with more overall rock cover (77%, SE = 3), wider entrances (3.1 cm, SE = 0.3), and access to a skyward-facing crevice, creating warmer microclimates. Our work indicates that rock features provide essential summer habitat for male little brown myotis and that lodgepole pine in this landscape may not provide appropriate roosting features. Understanding how other bat species may be using rock features, during summer and other seasons, remains a sizeable information gap. Learning more about hibernacula is of great importance due to the spread of white-nose syndrome and rock features may be essential autumn transitional roosts and winter hibernacula at higher elevations. Roosts that provide variation in microclimate, including the potential for passive warming, could be very beneficial for bats recovering from white-nose syndrome.Item Fanihi : a cultural digest. Cannibalism or conservation?(Montana State University - Bozeman, College of Arts & Architecture, 2010) Tharp, James Whitney; Chairperson, Graduate Committee: Ronald Tobias.There is a conflict on the island of Rota in the Commonwealth of the Northern Mariana Islands between conservationists and the cultural traditions of Chamorro inhabitants. The traditional model of broadcasting media to a wide audience is not effective in intervening in environmental conflicts within indigenous communities compared to an emerging model of filmmaking that embraces local voices and perspectives. Historically, indigenous depictions in media tend to misrepresent "Others" in order to reinforce the imperialist interests of Western society. Within this essay I intend to analyze how Western media suppresses indigenous voice while investigating strategies for the creation of effective environmental films targeted towards specific local audiences. Awareness of the mistakes of filmmakers of the past combined with the availability of inexpensive production and distribution technologies will allow alternative models of filmmaking to portray a diversity of perspectives. Environmental films that feature indigenous voices allow local communities to define and strengthen their own cultural values while creating texts that broaden global understandings of the diversity of the human experience.