abstract: Using a simple $SI$ infection model, I uncover the overall dynamics of the system and how they depend on the incidence function. I consider both an epidemic and endemic perspective of the model, but in both cases, three classes of incidence functions are identified. In the epidemic form, power incidences, where the infective portion $I^p$ has $p\in(0,1)$, cause unconditional host extinction, homogeneous incidences have host extinction for certain parameter constellations and host survival for others, and upper density-dependent incidences never cause host extinction. The case of non-extinction in upper density-dependent incidences extends to the case where a latent period is included. Using data from experiments with rhanavirus and salamanders, maximum likelihood estimates are applied to the data. With these estimates, I generate the corrected Akaike information criteria, which reward a low likelihood and punish the use of more parameters. This generates the Akaike weight, which is used to fit parameters to the data, and determine which incidence functions fit the data the best. From an endemic perspective, I observe that power incidences cause initial condition dependent host extinction for some parameter constellations and global stability for others, homogeneous incidences have host extinction for certain parameter constellations and host survival for others, and upper density-dependent incidences never cause host extinction. The dynamics when the incidence function is homogeneous are deeply explored. I expand the endemic considerations in the homogeneous case by adding a predator into the model. Using persistence theory, I show the conditions for the persistence of each of the predator, prey, and parasite species. Potential dynamics of the system include parasite mediated persistence of the predator, survival of the ecosystem at high initial predator levels and ecosystem collapse at low initial predator levels, persistence of all three species, and much more. / Dissertation/Thesis / Doctoral Dissertation Mathematics 2017

abstract: Epidemiological theory normally does not predict host extinction from infectious disease because of a host density threshold below which pathogens cannot persist. However, host extinction can occur when a biotic or abiotic pathogen reservoir allows for density-independent transmission. Amphibians are facing global population decline and extinction from the emerging infectious disease chytridiomycosis, caused by the fungus Batrachochytrium dentrobatidis (Bd). I use the model species Eleutherodactylus coqui to assess the impact of Bd on terrestrial direct-developing frog species, a common life history in the tropics. I tested the importance of two key factors that might influence this impact and then used laboratory experiments and published field data to model population-level impacts of Bd on E. coqui. First, I assessed the ontogenetic susceptibility of E. coqui by exposing juvenile and adult frogs to the same pathogen strain and dose. Juveniles exposed to Bd had significantly lower survival rates compared with control juveniles, while adult frogs often cleared infection. Second, I conducted experiments to determine whether E. coqui can become infected with Bd indirectly from contact with zoospores shed onto vegetation by an infected frog and from direct exposure to an infected frog. Both types of transmission were observed, making this the first demonstration that amphibians can become infected indirectly in non-aquatic habitats. Third, I tested the hypothesis that artificially-maintained cultures of Bd attenuate in pathogenicity, an effect known for other fungal pathogens. Comparing two cultures of the same Bd strain with different passage histories revealed reduced zoospore production and disease-induced mortality rates for a susceptible frog species (Atelopus zeteki) but not for the less-susceptible E. coqui. Finally, I used a mathematical model to project the population-level impacts of chytridiomycosis on E. coqui. Model analysis showed that indirect transmission, combined with either a high rate of zoospore production or low rate of zoospore mortality, is required for Bd to drive E. coqui populations below an extinction threshold. High rates of transmission plus frequent re-infection could lead to poor recruitment of infected juveniles and population decline. My research adds further insight into how emerging infectious disease is contributing to the loss of amphibian biodiversity. / Dissertation/Thesis / Ph.D. Biology 2013

Orientador: Luís Felipe de Toledo Ramos Pereira / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-25T14:12:31Z (GMT). No. of bitstreams: 1 Lambertini_Carolina_M.pdf: 6513639 bytes, checksum: 76f60b46e28d803521c2f06db67818b3 (MD5) Previous issue date: 2014 / Resumo: Os anfíbios são os animais mais ameaçados do planeta, tendo como fatores mais impactantes a perda de habitat e a quitridiomicose. Esta doença em anuros é causada pelo fungo Batrachochytrium dendrobatidis (Bd), que já foi registrado em todos os continentes e no Brasil estende-se por toda a Mata Atlântica, e já foi encontrado no Cerrado e na Amazônia. Existem diversos fatores que podem influenciar a dinâmica de infecção na natureza, e como fatores intrínsecos ao Bd podemos citar a variação genotípica, morfológica, variação na virulência e taxas de crescimento. Com isso, o presente estudo teve como objetivos realizar a caracterização genotípica e fenotípica de cepas de Bd provenientes de um gradiente de elevação na Mata Atlântica, e verificar se existem associações entre o tamanho dos zoósporos e zoosporângios das cepas isoladas com dados de prevalência e intensidade de infecção e taxa de crescimento do Bd. Adicionalmente, foi analisado se as taxas de infecção aumentam conforme o aumento da elevação. Para tanto, foram isoladas e genotipadas cinco cepas de Bd. Foi realizado o diagnóstico e quantificação da doença, assim como a confecção de suas curvas de crescimento. Com base nos resultados foram desenvolvidos dois índices relacionados à variação em tamanho e dados de prevalência e intensidade de infecção. Todas as cepas isoladas pertencem à linhagem Bd-GPL2. Foi detectada variação fenotípica entre as cepas e associações entre tamanho das cepas com prevalência e intensidade de infecção, e não houve associação com as taxas de crescimento. A prevalência e intensidade de infecção não aumentaram com a altitude. Os índices de tamanho e infecção também foram associados. Com isso, a variação fenotípica pode ser considerada preditora da dinâmica de infecção do Bd na natureza. Isto poderá ser utilizado como eficiente ferramenta de baixo custo para estudos que busquem reconhecer áreas nas quais o fungo pode ser mais prevalente e apresentar maior intensidade de infecção. Estes fatores podem ser cruciais para determinar populações de anfíbios ameaçadas / Abstract: Amphibians are the most threatened animals of the world. Habitat loss and chytridiomicosys are considered the main causes for their declines. Chytridiomycosis is a disease caused in anurans by the fungus Batrachochytrium dendrobatidis (Bd), detected worldwide. In Brazil it was registered all over the Atlantic rainforest and in some localities of the Cerrado and Amazonia. Several factors influence this disease dynamics, such as intrinsic factors related to Bd: different lineages, morphology, virulence, and growth rate. Given these factors, the present study aimed to genotype and analyze Bd strain¿s size from an elevational range, and verify if these factors were correlated to Bd size, prevalence, infection load, growth rate, and elevation. We isolated and genotyped five strains, and quantified growth rates. We developed two indexes related to Bd size, prevalence, and infection load. The five strains were Bd-GPL2. We found size variation and correlations between Bd size, prevalence, and infection load, but not with growth rate. Prevalence and infection load did not increase with elevation. The size and infection indexes were correlated. The phenotypic variation can be considered as a predictor to disease dynamics on environment. It can be used in other studies as a tool to efficiently (cheap and fast) identify areas where the fungus can develop faster and be more deleterious to amphibian populations / Mestrado / Biodiversidade Animal / Mestra em Biologia Animal

AN ABSTRACT OF THE DISSERTATION OF GRETCHEN ELIZABETH PADGETT-FLOHR for the Doctor of Philosophy degree in Zoology, presented on March 5, 2009 at Southern Illinois University of Carbondale. TITLE: BATRACHOCHYTRIUM DENDROBATIDIS IN CENTRAL CALIFORNIA AMPHIBIANS MAJOR PROFESSOR: Eric Schauber Amphibian chytridiomycosis has been identified as a disease responsible for the decline and extinction of many amphibian taxa world wide, but little research has been conducted on the disease in Mediterranean climates. To address this gap in the data I studied the amphibian assemblage present across a ~6,475 ha site in central California and investigated the occurrence of the etiological agent, Batrachochytrium dendrobatidis (BD) from organismal, community, landscape and historical perspectives. I initially tested the accuracy and reliability of a proposed diagnostic screening test for BD in four larval species that occur on the site. The screening test proposed by Fellers et al. (2001) and Vredenburg and Summers (2001) consisted of examining larval amphibian mouthparts for abnormalities and or defects, based on their hypothesis that mouthpart defects are clinical signs of BD infection. Sensitivity and specificity of the diagnostic screening test were 76% and 58%, respectively, indicating that the proposed screening test was not a reliable diagnostic test for BD infection for the four species I examined. I conducted controlled laboratory experiments to examine the consequences of BD infection in the two threatened California species that occur on my study site: Rana draytonii and Ambystoma californiense. Both species were susceptible to infection, but all infected animals survived the 18-month study. Infected A. californiense sloughed skin at three times the rate of uninfected salamanders, a pattern that may have long-term energetic costs potentially leading to population-level consequences of sublethal infection by BD. I conducted a retrospective survey of the California Academy of Sciences' (San Francisco, California, USA) amphibian collection, testing for BD in four amphibian species collected from central California between 1897 and 2005 to assess whether the pathogen is novel versus endemic. The earliest detection of BD was in two Rana catesbeiana collected in 1961, and the data support the hypothesis that BD was a novel pathogen introduced into central California prior to 1961 that spread geographically and taxonomically from at least one point of introduction and is now endemic throughout most of central California. I analyzed how environmental factors, amphibian community composition, land use practices, and landscape structure affect the dynamics of the pathogen's distribution on my study site in central California. The distribution of BD in ponds within the landscape varied markedly between years and increases were associated with precipitation, mean minimum and maximum temperatures, and presence of particular species. Pseudacris regilla infection patterns were highly indicative of overall patterns of pond BD status. Fourteen ponds were identified as BD hotpots (BD-positive three of four years). Occurrences of the pathogen within the landscape were spatially autocorrelated and ponds in close proximity to BD hotspots were more likely to test positive. Local land use, (presence/absence of grazing or recreational activity and developed lands), apparently did not influence BD status of a pond. My studies show that BD was likely a novel pathogen introduced into California ca. 1961 that has since become established as an endemic pathogen throughout most of central California. The listed amphibian species that occur in central California can be infected with BD but appear to be resistant to manifesting amphibian chytridiomycosis, and the data from the studies herein could support one of two hypotheses: that natural selection acting over the past 48 years has selected for those individuals that were resistant to the disease; or that the species on my site have always been resistant to BD. The research I conducted further supports the hypothesis that BD is locally vectored by native amphibians (e.g. P. regilla) moving between ponds and that local ecological constraints likely limit vectoring of BD by non-native species. These findings contribute substantially to elucidating and understanding the responses of amphibian populations to disease/pathogen introduction and lay groundwork for future investigations into the host-pathogen-environment relationship as it relates to declining amphibian populations.

<p>Infectious  diseases  are  becoming  increasingly  common  and  problematic  for  wildlife populations in many parts of the world. Disease prevalence and severity fluctuate over time, often due to the ubiquitous pressure of seasonality, or  the cyclic changes in ecological systems. However, for  many  host-pathogen  systems,  our  understanding of important  seasonal  drivers of  disease remains fragmented. For example, when seasonality of a disease is studied in medium and high latitudes, winter is often neglected, despite this being a period of physiological and immunological challenges associated with extreme environmental conditions. Therefore, my aim is to examine  drivers  of winter Infection dynamics of the amphibian fungal pathogen <em>Batrachochytrium dendrobatidis</em>. First, <em>B. dendrobatidis </em>infection prevalence was measured from spring through fall to understand local infection levels and anthropogenic influences (Chapter 1). Next, laboratory  experiments examined the effects of <em>B. dendrobatidis </em>on critical thermal minimum of two anuran species to identify how this pathogen influences the ability of hosts to survive winter (Chapter 2). Another laboratory experiment tested how road de-icing salt (commonly used in winter) influences <em>B. dendrobatids </em>and a tadpole host condition and subsequent host-pathogen interactions (Chapter 3). Finally, a field-based exploration of <em>B. dendrobatidis </em>infection dynamics was conducted during winter to understand how infections vary during in this season and between two overwintering strategies (Chapter  4). Winter  infection  dynamics  in  many  host-pathogen  systems are largely understudied, but the knowledge gained by this work can demonstrate how harsh environmental conditions of winter that can exacerbate otherwise benign infections, and affect the ability of hosts to sustain infections during winter. This work can therefore inform predictions and investigations of  infection  dynamics in  subsequent  seasons to  better  understand  the  seasonality  of  wildlife diseases.</p>

Batrachochytrium dendrobatidis (Bd) is a fungus that causes chytridiomycosis in amphibians. This fungus is an invasive species that has spread all over the world and is causing mass deaths in many areas. Bd has an efficient life cycle which targets suitable hosts and causes disturbances in amphibian osmoregulation and eventually death. It has caused more than 90 presumed extinctions worldwide. However, even though this fungus is commonly found in Sweden, no cases of chytridiomycosis has been found in the wild. This raises the questions i) what factors makes the infection in amphibians more severe and ii) does temperature affect the severity of Bd infection? According to the thermal mismatch hypothesis pathogens should be more efficient at temperatures where the performance gap between the host and pathogen is the widest. We chose the common toad as a study organism since it can be found all over Sweden and is generally considered to be adapted to colder climates. Our aim was to test the thermal mismatch hypothesis and see if the effect of infection differed between different temperatures. Common toad eggs were collected from two ponds in southern Sweden, raised in the laboratory and metamorphosed juvenile toads were reared in three different temperature rooms, either 14℃, 18℃ or 22℃. There they were either infected with Bd or exposed to a sham infection and monitored for 40 days. We show that Bd infection negatively affects growth and survival of common toads in accordance with previous studies. Furthermore, we found that temperature affects the effect of infection and shows patterns of the thermal mismatch hypothesis.

Emergent infectious diseases are a major driver of the accelerated rates of biodiversity loss that are being documented around the world. Global losses of amphibians provide evidence of this, especially those associated with chytridiomycosis, a lethal skin disease caused by the fungus Batrachochytrium dendrobatidis (Bd). Amphibian skin can harbor diverse bacterial communities that, in some cases, can inhibit the growth of Bd. Thus, there is interest in using skin bacteria as probiotics to mitigate Bd infections in amphibians. However, experiments testing this conservation approach have yielded mixed results, suggesting a lack of understanding about the ecology of these microbial communities. My dissertation research aimed to assess basic ecological questions in microbial ecology and to contribute to the development of probiotics using amphibian skin bacteria. First, to assess whether environmental conditions influence the function of amphibian skin bacterial communities, I conducted a field survey across low and high elevation populations of an amphibian host to assess their skin bacterial communities and metabolite profiles. I found that similar bacterial communities produced different metabolites at different locations, implying a potential functional plasticity. Second, since culturing is critical for characterizing bacteria, I aimed to identify the culture media (low vs high nutrient concentration) that recovers the most representative fraction of the amphibian skin bacterial community. I found that media with low nutrient concentrations cultured a higher diversity and recovered a more representative fraction of the diversity occurring on amphibian skin. I also determined that sampling more individuals is critical to maximize culture collections. Third, I assessed the diversity of the amphibian skin fungal community in relation to Bd infection across eight amphibian species. I determined that amphibian species was the most important predictor of fungal diversity and community structure, and that Bd infection did not have a strong impact. My dissertation highlights the importance of environmental conditions in the function of amphibian skin bacteria, expands our knowledge of the understudied fungal component of the amphibian skin microbiome, and complements current efforts in amphibian conservation. / Ph. D. / In light of the global losses of amphibian diversity due to, in part, the skin disease chytridiomycosis (caused by the fungus Batrachochytrium dendrobatidis [Bd]); the discovery that some amphibian-skin bacteria can inhibit Bd growth provides hope for amphibian conservation via their use as probiotics to control Bd infections. However, experiments testing these bacteria have yielded inconsistent results, suggesting a limited understanding about the factors influencing the diversity of amphibian-skin microbes and their ability to inhibit Bd. Also, efforts to identify effective candidates for probiotic therapy are still premature. Thus, my dissertation had an ecological emphasis and focused on complementing conservation efforts focused on probiotics. First, I assessed whether environmental conditions influence bacteriallyproduced products, which can have antifungal properties. Specifically, I surveyed low and highelevation populations of an amphibian species to assess the skin-bacteria and their products. I determined that, while skin bacterial communities were similar across an environmental gradient, their products differed, suggesting potential different antifungal properties. Second, I assessed the ability of different culture media types (low vs high nutrient concentrations) to grow a high portion and most representative fraction of the amphibian-skin bacteria. I found that culture media with low nutrient concentrations allowed the growth of a higher diversity of the bacteria occurring on the amphibian-skin, including the abundant members, and also determined that including a large number of amphibians is the best way to improve culture collections. Third, I assessed the fungal diversity occurring in the skin of different amphibian species and how it might response to Bd infections, and examined whether skin-fungi interact with co-occurring bacteria. I found that the amphibian species was the most important driver of the fungal diversity, and that Bd infection did not influence the diversity of these communities. Moreover, I identified the most diverse fungal phyla occurring in the amphibian-skin and determined that these fungi might interact with co-occurring bacteria. My dissertation contributes to our understanding about the influence of the environmental conditions in the amphibian-skin bacteria, expands our limited knowledge on the amphibian-skin fungi, and complement current amphibian conservation efforts.

No description available.

Depuis plusieurs décennies, un déclin important de la biodiversité est observé à l'échelle mondiale. Les amphibiens constituent aujourd'hui le groupe le plus vulnérable sur la planète. Près d'un tiers des espèces recensées dans le monde est à ce jour menacé d'extinction. Dans le cadre de ce doctorat, nous nous sommes intéressés dans un premier temps à l'émergence d'une maladie infectieuse des amphibiens, la chytridiomycose, provoquée par le champignon pathogène Batrachochytrium dendrobatidis (Bd). Nous avons étudié la répartition actuelle de ce champignon en France, démontré son impact sur la batrachofaune locale et mis en évidence le rôle de la Grenouille taureau (Lithobates catesbeianus) comme vecteur de transmission du pathogène. Afin de limiter la dissémination de Bd, nous avons également élaboré deux protocoles d'hygiène à mettre en oeuvre lors d'interventions sur le terrain ou lors d'utilisations d'amphibiens en laboratoire. Dans une seconde partie, nous avons développé une nouvelle méthode d'inventaire de la biodiversité basée sur la détection de l'ADN environnemental (ADNe). Nous avons mis en évidence que la persistance de l'ADNe dans un écosystème d'eau douce était d'environ 15 jours et que cette méthode innovante permettait d'améliorer la détection d'espèces exotiques envahissantes, comme la Grenouille taureau. Nous avons ensuite développé cette approche pour le suivi d'autres groupes taxonomiques (poissons, macro-invertébrés, chiroptères, etc.), dans des milieux différents et en utilisant notamment les technologies de séquençage nouvelle génération. / Since several decades, a significant decline in biodiversity is observed worldwide. Amphibians are now the most vulnerable group on the planet. Nearly a third of known species in the world is today threatened of extinction. Among many causes, diseases appear as an emerging threat worldwide. As part of this PhD, we were interested at first to the emergence of an infectious disease of amphibians, chytridiomycosis, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd). We studied the current distribution of this fungus mostly in France, showed the impact on local batrachofauna and highlighted the role of the Bullfrog (Lithobates catesbeianus) as a vector for transmission of the pathogen. To limit the spread of Bd, we also developed two hygiene protocols to implement during field trips or use of amphibians in the laboratory. In the second part of this thesis, we have developed a new method of biodiversity inventory based on the detection of environmental DNA (eDNA). We have shown that the persistence of vertebrates (fish and amphibian) eDNA in freshwater ecosystems was about 15 days and that this innovative method greatly improves the detection of invasive alien species, such as Bullfrog. We are then developed this approach for monitoring other taxonomic groups (fish, macro-invertebrates, bats, etc...), in various environments, taking advantage of bio-technological developments such as next generation DNA sequencing.

Chytridiomycosis is an emerging cutaneous fungal disease that contributes to recent global declines and extinction of amphibian species, caused by infection of the skin with a fungus known as Batrachochytrium dendrobatidis (Bd). Many species of frogs secrete antimicrobial peptides onto their skin that are capable of killing Bd. This thesis is an investigation of the effect of corticosteroids on cutaneous innate immunity in frogs, in the context of infection with Bd. The general hypothesis was that injections of glucocorticoids would impair the cutaneous synthesis of these antimicrobial peptides, thereby increasing susceptibility to Bd infection. The objective of the first experiment was to measure and compare gene expression levels of cutaneous AMP’s in frogs treated with glucocorticoids with sham-treated controls. Wild-caught Lithobates pipiens were acclimatized and administered either the corticosteroid methylprednisolone or saline every 48 hours. Norepinephrine-elicited cutaneous secretions were collected prior to the first injection of corticosteroid or saline, and then every 8 days for 40 days. Gene expression of the AMP’s brevinin and ranatuerin in the cutaneous secretions was quantified relative to the reference genes EF1-α and RPL8 using reverse transcription quantitative polymerase chain reaction (RT-qPCR). Corticosteroid treatment was associated with a significant (P<0.027) increase in brevinin gene expression, which was most notable at 24-40 days of corticosteroid administration. Ranatuerin expression followed a similar but nonsignificant trend. The second experiment was a pilot study intended to establish a Bd challenge protocol in L. pipiens. Frogs were immersed in water containing 0, 104, 105 or 106 zoospores of Bd strain JEL 423. Cutaneous swabs were collected prior to challenge and tested for Bd by qPCR; unexpectedly, some tested positive, indicating pre-challenge infection. The analysis was complicated by an identified cross-reactivity of the assay with other fungi. The findings of the first experiment refuted the hypothesis, and suggested that corticosteroids promote rather than impair AMP gene expression in the skin of L. pipiens, under these experimental conditions. Further, the second study demonstrated that none of the frogs showed clinical abnormalities or died, despite exposure to Bd zoospores and despite molecular and histologic evidence of cutaneous Bd infection in some frogs. / NSERC Discovery Grant