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The bacteria in host-associated microbial communities influence host health through various mechanisms, such as immune stimulation or the release of metabolites. However, viruses that target bacteria, called bacteriophages (phages), may also shape the animal microbiome. Most phage lifecycles can be classified as either lytic or temperate. Lytic phages infect and directly kill bacterial hosts and can directly regulate bacterial population size. Temperate phages, in contrast, have the potential to undergo either a lytic cycle or integrate into the bacterial genome as a prophage. As a prophage, the phage may alter bacterial host phenotypes by carrying novel genes associated with auxiliary metabolic functions, virulence-enhancing toxins, or resistance to other phage infections. Lytic phages may also carry certain auxiliary metabolic genes, which are instead used to takeover bacterial host functions to better accommodate the lytic lifecycle. In either case, the ability to alter bacterial phenotypes may have important ramifications on host-associated communities. This dissertation focused on the genetic contributions that phages, and particularly prophages, provide to the bacterial members of two separate host-associated communities: the honey bee (Apis mellifera) gut microbiome and the amphibian skin microbiome. My second chapter surveyed publicly available whole genome sequences of common honey bee gut bacterial species for prophages. It revealed that prophage distribution varied by bacterial host, and that the most common auxiliary metabolic genes were associated with carbohydrate metabolism. In chapter three, this bioinformatic pipeline was applied to the amphibian skin microbiome. Prophages were identified in whole genome bacterial sequences of bacteria isolated from the skin of American bullfrogs (Lithobates catesbeianus), eastern newts (Notophthalmus viridescens), Spring peepers (Pseudacris crucifer) and American toads (Anaxyrus americanus). Prophages were additionally identified in publicly available genomes of non-amphibian isolates of Janthinobacterium lividum, a bacteria found both on amphibian skin and broadly in the environment. In addition to a diverse set of predicted prophages across amphibian bacterial isolates, several Janthinobacterium lividum prophages from both amphibian and environmental isolates appear to encode a chitinase-like gene undergoing strong purifying selection within the bacterial host. While identifying the specific function of this gene would require in vitro isolation and testing, its high homology to chitinase and endolysins suggest it may be involved in the breakdown of either fungal or bacterial cellular wall components. Finally, my fourth chapter revisits the honey bee gut system by investigating the role of geographic distance in bacteriophage community similarity. A total of 12 apiaries across a transect of the United States, from Virginia to Washington, were sampled and honey bee viromes were sequenced, focusing on the lytic and actively lysing temperate community of phages. Although each apiary possessed many unique bacteriophages, apiaries that were closer together did have more similar communities. Each bacteriophage community also carried auxiliary carbohydrate genes, especially those associated with sucrose degradation, and antimicrobial resistance genes. Combined, the results of these three studies suggest that bacteriophages, and particularly prophages, may be contributing to the genetic diversity of the bacterial community through nuanced relationships with their bacterial hosts. / Doctor of Philosophy / The microbial communities of animals, called "microbiomes", play important roles in the health of animals. The bacteria in these microbiomes can help strengthen the immune system, provide resistance to dangerous pathogens, and break down nutrients. However, bacteria are not alone in the microbiome; viruses are also present. Surprisingly, the vast majority of the world's viruses, even those living inside animals, infect bacteria. These viruses, called "bacteriophages" or "phages", can impact the bacterial communities in a microbiome. Phages can be grouped in to two broad categories based on lifecycle. Lytic phages kill the bacterial host directly after infection. Temperate phages, on the other hand, can either immediately kill the host like lytic phages or alternatively, become a part of the bacterial genome and live as prophages. Phages with both lifecycles can sometimes carry genes that, although not essential to the phage, may change the traits of the bacteria during infection. For example, some phages carry toxin genes, which bacteria use to cause disease in animals. Other phages might carry genes that provide antibiotic resistance or alter the metabolism of the infected bacteria. If a phage gene benefits the infected bacteria, the bacteria may begin interacting with its environment in a new way or may even become more abundant. Alternatively, phages that directly kill infected bacteria may have a negative effect on bacterial population sizes. To begin unraveling how phages influence bacterial species in microbiomes, I investigated two different animal systems: the Western honey bee (Apis mellifera) gut microbiome and the amphibian skin microbiome. I first identified prophages of several common bacterial species that reside in the honey bee gut (Chapter 2). Prophages were more common in certain bacterial species than others, and some possessed genes associated with the breakdown of sugars or pollen, suggesting they help honey bees process their food. Using similar techniques, I then identified prophages in bacteria isolated from the skin microbiomes of several amphibian species common in the eastern United States (American bullfrogs, Eastern newts, Spring peepers, and American toads) (Chapter 3). Most notably, the bacteria Janthinobacterium lividum may benefit from prophages that carry genes for potentially antifungal chitinase enzymes that destroy the fungal cell wall. Finally, I returned to the honey bee gut microbiome system by investigating how honey bee bacteriophage communities change over large geographic distances (Chapter 4). This study, which examined honey bees from 12 apiaries sampled from the east to west coast of the United States, looks primarily at lytic phage and temperate phage that are not integrated as prophage, but are instead seeking a bacterial host to infect. I found that nearby apiaries tended to have more similar communities of bacteriophages, compared to apiaries far away. Additionally, most bacteriophage communities carry genes associated with the breakdown of sugars like sucrose. Overall, these three studies show that phages, and especially prophages, contribute to the genetic landscape of the microbiome by broadly providing bacterial hosts with access to a diverse set of genes.

Current literature suggests that little, if any, research has been conducted in South Africa to determine the impact of alien invasive trout on indigenous amphibian biodiversity. The primary aim of the research project was to establish whether waterfalls are seasonally important in protecting the indigenous Natal Cascade Frog Hadromophryne natalensis tadpole populations from predation by alien rainbow trout Oncorhynchus mykiss and brown trout Salmo trutta at two sampling areas located at Injesuthi and Monk‟s Cowl Nature Reserve within the uKhahlamba Drakensberg Park World Heritage Site (UDPWHS). The dissertation argues in favour of a biotic rationale, namely trout predation, as being the primary cause for the decline of H. natalensis tadpoles below the waterfalls and systematically negates the influence of geo-physical (abiotic) environmental factors on tadpole abundance. Habitat isolation and fragmentation is identified as a latent threat to the continued persistence of H. natalensis populations in the UDPWHS. / Environmental Sciences / M. Sc. (Environmental Science)

Les plantes envahissantes sont considérées comme l'une des plus grandes menaces sur les espèces, mais leurs impacts sur les amphibiens demeurent peu connus. L’objectif de ce projet est de déterminer l’effet de l’établissement du roseau commun (Phragmites australis) sur la répartition des amphibiens et sur leur développement larvaire. Il est présumé que cette plante s'accapare l'espace et les ressources disponibles en produisant une importante biomasse, qu’elle peut modifier l’hydrologie des marais et la structure des communautés d’amphibiens. J’ai évalué les facteurs influençant la répartition des amphibiens selon les caractéristiques des étangs et du paysage dans 50 sites envahis ou non à divers degrés. Des expériences ont également été menées afin d’étudier les répercussions de trois densités de roseau sur des têtards de la Grenouille des bois (Lithobates sylvaticus) et sur la qualité de son habitat. Mes résultats suggèrent que le roseau à forte densité ralentit le développement larvaire de la grenouille des bois et influence les assemblages phytoplanctoniques. Cependant, il n’y a aucune relation entre la densité de la plante et la survie, la morphologie des têtards et les caractéristiques biotiques et abiotiques de l’eau. Dans notre aire d’étude, le paysage autour des étangs a une plus grande influence sur la répartition des amphibiens que l'établissement du roseau. Toutefois, la probabilité d’assèchement est plus élevée lorsque la plante est établie en grande quantité, ce qui, si l’envahissement s’intensifie, aura un effet néfaste sur la survie des têtards et mettra en péril la persistance des populations. / Invasive plants are considered one of the greatest threats to species, but their impact on amphibians is still poorly understood. The objective of this project is to determine the effect of the establishment of common reed (Phragmites australis) on amphibian distribution and larval development. It is thought that this plant monopolizes space and resources by producing a large biomass, and may alter wetland hydrology and amphibian community structure. I evaluated the factors influencing amphibian distribution according to the characteristics of ponds and the surrounding landscape in 50 sites invaded or not by reeds to varying degrees. Experiments were also conducted to study the impacts of three reed densities on wood frog tadpoles (Lithobates sylvaticus) and the quality of their habitat. My results suggest that high reed density slows wood frog larval development and influences phytoplankton assemblages. However, there is no relationship between, plant density and survival, tadpole morphology and water biotic and abiotic characteristic. In our study area, the landscape surrounding ponds has a greater influence on amphibian distribution than does reed establishment. However, the desiccation probability is higher when the plant is established in high quantities, which, if the invasion intensifies, will have an adverse effect on tadpole survival and therefore population persistence.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Na Colômbia ocorrem 787 espécies de anfíbios. Por causa da preocupação com o estado de conservação de muitas dessas espécies, tem se sugerido que na Colômbia deveriam-se priorizar as pesquisas em taxonomia e ecologia em regiões sub-amostradas a fim de intensificar o conhecimento e conservação dos anfíbios colombianos. Baseados em uma análise cienciométrica de 319 trabalhos sobre a ecologia dos anfíbios colombianos publicados entre 1840 e 2014 (No Capítulo 1), identificamos as tendências nos esforços realizados em distintos temas de pesquisa, e a distribuição regional e taxonômica desses estudos. A maioria dos estudos (67%) foi realizada na região Andina colombiana em comparação com outras regiões naturais da Colômbia. Apenas 46% das espécies de anfíbios ocorrendo na Colômbia foi tratada nos estudos analizados, e a maioria (58%) delas é da região Andina. Entre as publicações analizadas identificamos 14 temas de pesquisa em ecologia, dos quais ecologia reprodutiva (26%), conservação de espécies (23%) e dieta (14%) foram os mais pesquisados. Nossos dados mostraram que na Colômbia há um considerável avanço na pesquisa sobre a ecologia dos anfíbios do país, mas ainda são necessários esforços para cobrir muitos vazios de informação para muitas regiões e para muitas espécies de anfíbios que possuem dados incipientes. No sudoeste da Cordilheira Ocidental colombiana há pouca informação ecológica sobre os anfíbios ali ocorrendo. A fim de saber alguns aspectos ecológicos dessas espécies, desenvolvimos três estudos sobre a diversidade e ecologia de anfíbios presentes na Reserva Natural Río Ñambí (a seguir RNRÑ). No Capítulo 2 apresentamos uma análise sistemática do gênero Andinophryne (Família Bufonidae), composto por três espécies, A. atelopoides, A. colomai (presente na RNRÑ) e A. olallai. As filogenias mostraram que Andinophryne está incorporado dentro de Rhaebo. Portanto, sinonimizamos Andinophryne sob Rhaebo e discutimos as sinapomorfias morfológicas putativas para Rhaebo. Além, fornecemos informações ecológicas e sobre o estado de conservação das três espécies incluídas na nova combinação taxonômica. No Capítulo 3 apresentamos uma lista de 19 espécies de anfíbios pertencentes a oito famílias, com uma dominância numérica da família Craugastoridae e do gênero Pristimantis. As espécies com a maior abundância relativa (> 25%) foram Pristimantis labiosus e P. verecundus. Sete diferentes modos de reprodução foram reconhecidos, com a maioria das espécies (68%) possuindo desenvolvimento direto de ovos. Cinco (26%) das espécies registradas estão classificadas dentro das categorias de maior ameaça de extinção. Reportamos para sete espécies a extensão da faixa de distribuição geográfica latitudinal na Colômbia. No Capítulo 4 comparamos a dieta de jovens e adultos de P. labiosus para identificar se houve uma mudança ontogenética no tamanho de presa consumido com o aumento na largura da boca. A dieta foi composta por 19 categorias de presas (> artrópodes), com as duas classes de idade consumindo um similar espectro de categorias. Os jovens têm um nicho trófico maior (0,45) do que os adultos (0,25), com uma sobreposição de nicho relativamente baixa (0,39) entre eles. Apesar da diferencia na largura da boca entre jovens e adultos, não houve uma correspondente mudança ontogenética no tamanho de presa consumida. Consideramos P. labiosus como um predador generalista que parece consumer uma ampla gama de tipos e tamanhos de presas / Colombia harbors 787 species of amphibians. Because of concerns with the conservation status of many of these species, it has been suggested that in Colombia should prioritize research in taxonomy and ecology in undersampled regions in order to enhance the knowledge and conservation of Colombian amphibians. Based on a scientometric analysis of 319 works on ecology of Colombian amphibians published between 1840 and 2014 (Chapter 1), we identified trends in the efforts made in different research topics, and both regional and taxonomic distribution of these studies. Most studies (67%) was conducted in the Colombian Andean region compared to other natural regions of Colombia. Only 46% of amphibian species occurring in Colombia was included in the reviewed studies, and the most of them (58%) is from the Andean region. Among the reviewed publications, we identified 14 research topics in ecology, of which reproductive ecology (26%), species conservation (23%), and diet (14%) were the most investigated. Our data showed that in Colombia, there is a considerable advance in research on amphibian ecology, but we need further efforts to fill many gaps in information for many regions and for many amphibian species possessing incipient data. In southwestern of the Colombian western Andes there is little information about the species occurring there. In order to know some ecological aspects of these species, we conducted three studies on diversity and ecology of the amphibians present in the Reserva Natural Río Ñambí (hereafter RNRÑ). In the Chapter 2, we show an analysis of the systematics of the genus Andinophryne (Family Bufonidae), composed by three species, A. atelopoides, A. colomai (present in the RNRÑ) and A. olallai. The phylogenies showed that Andinophryne is embedded within Rhaebo. Therefore, we synonymize Andinophryne under Rhaebo and we discussed the putative morphological synapomorphies for Rhaebo. In addition, we provide ecological informations and on the conservation status of the three species included in the new taxonomic combination. In the Chapter 3, we show a list of 19 amphibian species from eight families, with a numerical dominance of the Family Craugastoridae and of the genus Pristimantis. The species with the highest relative abundance (>25%) were Pristimantis labiosus and P. verecundus. Seven different reproductive modes were recognized, with most species (68%) presented direct development of eggs. Five (26%) of the species recorded are classified in the categories of greatest threat of extinction. We report for seven species an extension of the latitudinal distribution range in Colombia. In the Chapter 4, we compared the diet of juveniles and adults of P. labiosus in order to identify if there was an ontogenetic shift in the prey size consumed as mouth width increases. The diet was composed of 19 prey categories (> arthropods), and the two age-classes consumed a similar array of prey categories. Juveniles had a broader trophic niche (0.57) than adults (0.25), with a relatively low trophic overlap among them (0.39). Although we found differences in mouth width between juveniles and adults, we did not observe a corresponding ontogenetic shift in prey size consumption. We consider that P. labiosus is a generalist predator, consuming a wide range of prey types and sizes

Surfactants are one of the more ubiquitous contaminants in aquatic systems. Their importance as toxic components of pesticide formulations has, however, been largely overlooked. Amphibians particularly, as inhabitants of shallow, temporary and often lentic aquatic environments may be at risk from exposure to these chemicals when they enter aquatic systems. This thesis presents data on the toxicity of surfactants to amphibians. Several experimental exposures were conducted with embryo-larval, tadpole and adult developmental stages of the Australian species- Crinia insignifera, Helcioporus eyrei, Limnodynastes dorsalis and Litoria moorei and the exotic species- Bufo marinus and Xenopus laevis. Animals were variously exposed to glyphosate formulations that contain a high proportion of nonionic surfactants, or commercial pesticide wetting agents (alcohol alkoxylate and nonylphenol ethoxylate (NPE) surfactants). Feeding stage tadpoles of C.insignifera, H. eyrei, L. dorsalis and L. moorei were exposed to three commercial glyphosate formulations, glyphosate isopropylamine and glyphosate acid in static-renewal acute toxicity tests. The 48-h LC50 values for Roundup Herbicide (MON 2139) tested against tadpoles of C. insignifera, H. eyrei, L. dorsalis and L. moorei ranged between 8.1 and 32.2 mg/L (2.9 and 11.6 mg/L glyphosate acid equivalent (ae)), while the 48-h LC50 values for Roundup Herbicide tested against adult and newly metamorphosed C. insignifera ranged from 137-144 mg/L (49.4-51.8 mg/L ae). / Touchdown Herbicide (4 LC-E) tested against tadpoles of C. insignifera, H. eyrei, L. dorsalis and L. moorei was slightly less toxic than Roundup with 48-h LC50 values ranging between 27.3 and 48.7 mg/L (9.0 and 16.1 mg/L ae). Roundup Biactive (MON 77920) was practically non-toxic to tadpoles of the same four species producing 48-h LC50 values of 911 mg/L (328 mg/L ae) for L. moorei and >1000 mg/L (>360 mg/L ae) for C. insignifera, H. eyrei and L. dorsalis. Glyphosate isopropylamine was practically non-toxic producing no mortality amongst tadpoles of any of the four species over 48 h, at concentrations between 503 and 684 mg/L (343 and 466 mg/L ae). The toxicity of technical grade glyphosate acid (48-h LC50, 81.2-121 mg/L) is likely to be due to acid intolerance. Feeding stage tadpoles of B. marinus, X laevis, C. insignifera, H.eyrei, L. dorsalis and L. moorei were exposed to NPE and alcohol alkoxylate in static renewal acute toxicity tests. All species exhibited non-specific narcosis following exposure to both these surfactants. The 48-h EC50 values for NPE ranged between 1.1 mg/L (mild narcosis) and 12.1 mg/L (full narcosis). The 48-h EC50 values for alcohol alkoxylate ranged between 5.3 mg/L (mild narcosis) and 25.4 mg/L (full narcosis). Xenopus laevis was the most sensitive species tested. The sensitivity of the other five species was size dependent with larger species displaying greater tolerance. Replicate acute toxicity tests with B. marinus exposed to NPE at 30 degrees celsius over 96 hours indicated that the narcotic effects were not particularly time dependant. / The mean 24, 48, 72 and 96-h EC50 (mild narcosis) were 3.6, 3.7, 3.5 and 3.5 mg/L respectively. The mean 24, 48, 72, and 96-h EC50 (full narcosis) values were 4.0, 4.1, 4.2 and 4.0 respectively. Acute toxicity tests with B. marinus exposed to NPE at 30 degrees celsius under conditions of low dissolved oxygen (0.8-2.3 mg/L) produced a two to threefold increase in toxicity. The 12-h EC50 values ranged from 1.4 to 2.2 mg/L. The embryotoxicity of NPE was determined in X. laevis, L. adelaidensis and C. insignifera using a Frog Embryo Teratogenesis assay-Xenopus (FETAX). The 96-h LC50, EC50 and MCIG (LOEC) values for X. laevis were 3.9 to 5.4 mg/L, 2.8 to 4.6 mg/L and 1.0 to 3.0 mg/L respectively. The 140-h LC50, ECSO and MCIG values for L. adelaidensis were 9.2 mg/L, 8.8 mg/L and 5.1 to 6.0 ing/L respectively. The 134-h LC50, EC50 and MCIG values for C. insignifera were 6.4 mg/L, 4.5 mg/L and 4.0 mg/L respectively. Teratogenicity indices for the three species ranged between 1.0 and 1.6 indicating either no or low teratogenicity. Xenopus laevis was the more sensitive of the three species and the only species that displayed indisputable terata. The acute toxicity data indicated that the amphibian species tested were of similar sensitivity to fish and some invertebrates. Developmental retardation and oestrogenic effects following exposure to nonylphenol ethoxylate were indicated by sublethal toxicity tests. Crinia insignifera embryos were exposed during early embryogenesis to sublethal concentrations of NPE. / Exposure to NPE did not affect either weight nor size (snout-vent length) at metamorphosis. Exposure to 5.0mg/L NPE resulted in a significant delay in the time required to reach metamorphosis. Also, exposure to 3.0 mg/L NPE for the first 6 days of embryonic development or exposure to 5.0 mg/L NPE from day 2 to day 6 resulted in a statistically significant predominance in the female phenotype amongst metamorphosing froglets. Exposure for the first five days to 1.5 ing/L or 3.0 mg/L NPE had no effect on sex ratio. The results indicated that exposure to NPEs has endocrine disruptive effects in this species and that a narrow window of susceptibility exists for the induction of predominantly female phenotype. This study has also followed the degradation of a mixture of NPE oligomers and the concomitant formation of individual oligomers in static die-away tests with and without illumination in freshwater. Over 33 days in darkness there was a progressive and complete loss of long chain oligomers (NPEO(subscript)8-17), transient increases and subsequent loss of short to medium chain oligomers (NPE0(subscript)4-7), and large persistent increases (approximately 1000%) in short chain oligomers (NPE0(subscript)1-3). In the presence of illumination, biodegradation was retarded and heterotrophic bacterial proliferation was inhibited. After 33 days there was complete loss of long chain oligomers (NPE0(subscript)9-17), incomplete loss of medium chain oligomers (NPE0(subscript)6.8) and increases in short chain oligomers (NPE0(subscript)1-5). / This thesis discusses the importance of persistent metabolites of NPE degradation as it pertains to the habitat, developmental time frame and ecology of amphibians. Degradation of NPE is likely to occur over a time frame that is longer than that required for complete embryogenesis and metamorphosis of many species of amphibians, and may easily encompass those critical stages of development during which oestrogenic metabolites can affect development.

Les plantes envahissantes sont considérées comme l'une des plus grandes menaces sur les espèces, mais leurs impacts sur les amphibiens demeurent peu connus. L’objectif de ce projet est de déterminer l’effet de l’établissement du roseau commun (Phragmites australis) sur la répartition des amphibiens et sur leur développement larvaire. Il est présumé que cette plante s'accapare l'espace et les ressources disponibles en produisant une importante biomasse, qu’elle peut modifier l’hydrologie des marais et la structure des communautés d’amphibiens. J’ai évalué les facteurs influençant la répartition des amphibiens selon les caractéristiques des étangs et du paysage dans 50 sites envahis ou non à divers degrés. Des expériences ont également été menées afin d’étudier les répercussions de trois densités de roseau sur des têtards de la Grenouille des bois (Lithobates sylvaticus) et sur la qualité de son habitat. Mes résultats suggèrent que le roseau à forte densité ralentit le développement larvaire de la grenouille des bois et influence les assemblages phytoplanctoniques. Cependant, il n’y a aucune relation entre la densité de la plante et la survie, la morphologie des têtards et les caractéristiques biotiques et abiotiques de l’eau. Dans notre aire d’étude, le paysage autour des étangs a une plus grande influence sur la répartition des amphibiens que l'établissement du roseau. Toutefois, la probabilité d’assèchement est plus élevée lorsque la plante est établie en grande quantité, ce qui, si l’envahissement s’intensifie, aura un effet néfaste sur la survie des têtards et mettra en péril la persistance des populations. / Invasive plants are considered one of the greatest threats to species, but their impact on amphibians is still poorly understood. The objective of this project is to determine the effect of the establishment of common reed (Phragmites australis) on amphibian distribution and larval development. It is thought that this plant monopolizes space and resources by producing a large biomass, and may alter wetland hydrology and amphibian community structure. I evaluated the factors influencing amphibian distribution according to the characteristics of ponds and the surrounding landscape in 50 sites invaded or not by reeds to varying degrees. Experiments were also conducted to study the impacts of three reed densities on wood frog tadpoles (Lithobates sylvaticus) and the quality of their habitat. My results suggest that high reed density slows wood frog larval development and influences phytoplankton assemblages. However, there is no relationship between, plant density and survival, tadpole morphology and water biotic and abiotic characteristic. In our study area, the landscape surrounding ponds has a greater influence on amphibian distribution than does reed establishment. However, the desiccation probability is higher when the plant is established in high quantities, which, if the invasion intensifies, will have an adverse effect on tadpole survival and therefore population persistence.

L’eugénol permet d’induire une anesthésie chirurgicale chez la grenouille africaine à griffes (Xenopus laevis) sans causer de lésions chez des grosses grenouilles (90-140g). Le premier objectif de la présente étude était de déterminer la durée de l’anesthésie et d’évaluer la dépression du système nerveux central ainsi que les changements de saturation en oxygène et de fréquence cardiaque chez des petites (7.5 ± 2.1 g) et moyennes (29.2 ± 7.4 g) grenouilles Xenopus laevis en fonction du temps d’exposition à un bain d’eugénol de 350 µL/L. Suite à une immersion de 5 ou 10 minutes, la réponse au test à l’acide acétique, au réflexe de retrait et au réflexe de retournement était absente pendant 1 heure (petites grenouilles) et 0,5 heure (moyennes) et l’anesthésie chirurgicale durait au maximum 15 et 30 minutes chez les petites et moyennes grenouilles respectivement. La saturation en oxygène n’était pas affectée de façon significative, mais la fréquence cardiaque était diminuée jusqu’à 1 heure post-immersion dans les deux groupes. Le deuxième objectif était de déterminer la toxicité de l’eugénol chez des grenouilles de taille moyenne après une ou trois administrations à une dose anesthésique, avec ou sans période de récupération d’une semaine. Histologiquement, il y avait de l’apoptose tubulaire rénale et des membranes hyalines pulmonaires après une administration, et de la nécrose hépatique et des hémorragies dans les tissus adipeux après trois administrations. Ces résultats suggèrent que le poids corporel est un paramètre important à considérer lors de l’anesthésie de grenouilles Xenopus laevis par immersion dans l’eugénol. / Eugenol has been shown to induce surgical anesthesia in African clawed frogs (Xenopus laevis) without causing lesions after a single administration in large frogs (90-140g). The first objective of this study was to determine the duration of anesthesia in small (7.5 ± 2.1 g) and medium (29.2 ± 7.4 g) Xenopus laevis frogs and evaluate CNS depression and changes in oxygen saturation and heart rate relative to exposure time in a eugenol bath (350 µL/L). After immersion for 5 or 10 minutes, no responses to the acetic acid test (AAT), withdrawal reflex, and righting reflex were seen for 1 h (small frogs) or 0.5 h (medium frogs), and small and medium frogs were under surgical anesthesia for a maximum of 15 and 30 minutes respectively. Oxygen saturation was not significantly affected by anesthesia, but heart rate was depressed for as long as 1 hour post-exposure in both groups of frogs. The second objective was to determine the toxicity of eugenol in medium frogs after one or three administrations at anesthetic doses, with or without a 1 week recovery period. Histopathology revealed renal tubular apoptosis and pulmonary hyaline membranes after 1 administration, as well as hepatic necrosis and adipose tissue hemorrhages after 3 administrations. These results suggest that body weight is an important parameter to consider when using a eugenol bath for anesthesia of Xenopus laevis frogs.

L'écotoxicité de différentes nanoparticules de carbone (NPC) a été évaluée chez des organismes aquatiques, en particulier chez Xenopus laevis. Il a été montré que la surface des NPC est le paramètre le plus pertinent pour décrire l'inhibition de croissance chez le xénope, indépendamment de leur forme allotropique et de leur état de dispersion. L'induction des micronoyaux a aussi été étudiée chez le xénope, et l'oxyde de graphène (GO) s'est révélé génotoxique à faible dose, résultat corroboré par l'étude de l'expression des gènes. Les mécanismes de toxicité impliqués seraient notamment liés aux fonctions oxygénées de la particule. De plus, le GO a aussi entrainé de la génotoxicité chez Pleurodeles waltl. et de la tératogénicité, des retards de développement et de l'inhibition de croissance chez Chironomus riparius. La mise en interaction de ces organismes au sein d'un mésocosme a également conduit à l'observation de génotoxicité chez le pleurodèle en présence de GO. / The ecotoxicity of different carbon-based nanoparticles (CNPs) was assessed in freshwater organisms, especially in Xenopus laevis. The surface of the CNPs was shown to be the more relevant parameter to describe the growth inhibition in Xenopus, regardless of their allotropic form and their state of dispersion. Micronucleus induction was also studied in Xenopus and graphene oxide (GO) was found genotoxic at low dose. This result was in compliance with the study of genes expression. The involved toxicity mechanisms would be related to the oxidized functions of the CNP. Moreover, GO was also found responsible for genotoxicity in Pleurodeles waltl. and for teratogenicity, development delay and growth inhibition in Chironomus riparius. These organisms have finally been put together in a mesocosm, which has also led to genotoxicity in Pleurodeles in the presence of GO.