Environmental fluctuations in the eastern Pacific Ocean are reflected in the tissues of some of its most vulnerable apex predators, the Peruvian fur seal (PFS) Arctocephalus australis ssp.and the South American sea lion (SASL) Otaria byronia.These large pinnipeds live in sympatry along the Pacific coastline of South America and forage within the neritic waters over the continental shelf. The coastal waters off Peru are a region of great environmental fluctuations due to periodic, oscillating El Niño- La Niña- Southern Oscillation (ENSO) events, which result in ecosystem-wide food web changes. Pinniped vibrissae (whiskers) are continuously growing keratinous tissues and reflect the incorporation of prey from the region. Fine resolution sampling along their length provided trophic information on a weekly to monthly time scale over several years. Vibrissae were obtained from 2010-2016 from female (n=47), male (n=17) and newborn pup (n=6) fur seals and male (n=6) sea lions collected during pinniped rookery health assessments. Stable isotope ratios (δ13C and δ15N) in vibrissae infer temporal primary production and dietary variations in individuals. ENSO conditions were dictated by the sea surface temperature anomaly (SSTA) recordings from the Niño 1+2 Index region over 12 years. Fluctuations in vibrissae δ15N were correlated to varying SSTA readings (p15N (p15N signatures (p13C revealed minor fluctuations ranging from -18.13 to -13.17‰over the 12-year period, δ13C did reveal a significant depletion and enrichment oscillation during specific points in time, such as the 2014-2016 El Niño event, which ranged from -15.09 to -13.83‰(p=0.040). Stable isotope signatures varied between genders, but not by species. Female fur seal stable isotope signatures were significantly more depleted in both δ13C and δ15N than males (p13C and δ15N were inversely correlated to each other from 2014 to 2016; this was during the strongest El Nino-Southern Oscillation event on record (p=0.002). As δ13Csignatures became more enriched, δ15N signatures depleted from 2014 until 2016. This suggests that when ENSO warm phase conditions occur, the environments resources change in historic foraging grounds, possibly forcing pinnipeds to travel farther distances offshoreto forage or altering what they feed on throughout these stressful conditions.This study serves to help understand of the impact of oceanographic influences on these pinnipeds.ENSO conditions influence the trophic dynamics and resultant survival adaptations in both of these vulnerable Peruvian marine mammals.

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Craniometrics is a very reliable and effective tool for studying the difference in animal morphology. Previously, traditional craniometrics were conducted with the aid of calipers in two dimensional format (2D). Such discounting of actual three-dimensional 3D form may result in loss of some relevant and critical information leading to compromised and unreliable results for studies such as population variation analysis of morphology. The employment of 3D photogrammetry allows a close to complete representation of the physical dimensions of a specimen. The use of photogrammetry in mammalogy concentrated on measuring of body size/mass, but little has been done on animal skull delineation through photogrammetry. This dissertation describes advances in morphometrics and 3D photogrammetry application in craniometrics, investigates the craniometric variation between closely related species (Arctocephalus gazella and A. tropicalis), and A. tropicalis interpopulation craniometrics between two geographically distinct populations, at Marion Island and Gough Island, using Photomodeler Scanner® (PMSc®) three-dimensional (3D) modelling software to produce accurate, high resolution 3D skull models. A total of 117 3D models were created from adult male fur seal crania, and 16 traditional measurements recorded, using specimens archived at the Port Elizabeth Museum, Bayworld, South Africa. Sixteen linear measurements, (8 caliper recordings and 8 3D recordings) were used for PMSc® methodology testing, 16 (A. gazella n= 8 and A. tropicalis n= 8) used for species cranial comparison and 85 (Marion Island n= 54 and Gough Island n= 31) used for interpopulation variation. The craniometric variations were analysed using the Statistica® v11 software package, StatSoft, Inc. The comparison between linear traditional caliper cranial measurements and 3D measurements produced significantly similar results, attesting to the accuracy of the PMSc® 3D model production. Photomodeler Scanner® therefore produces accurate and high resolution 3D models of skulls which allow 3D measurements. I predicted that PMSc® would detect the existing significant differences between the skulls of adult male A. gazella and A. tropicalis and modelled and compared their 3D models, and I further predicted that PMSc® would detect any existing differences between the skulls of A. tropicalis from Gough and Marion islands by comparison of their 3D models. The Gough Island and Marion Island A. tropicalis populations could not be discriminated based on linear 3D cranial I conclude that PMSc® is a reliable and effective tool for accurate and high resolution 3D modelling. The present study confirms previous findings and contributes additional evidence that suggests that adult A. tropicalis males from Gough Island and Marion Island cannot be discriminated based on linear measurements of craniometrics, and deviates from the Bergmanian rule as applied to large mammals. The present study, however, makes several noteworthy contributions to the use of PMSc® and 3D modelling in morphometrics. Taken together, these findings suggest a role for PMSc® 3D modelling in promoting accurate digitization of museum specimens and creation of online museum libraries. This research will serve as a baseline for future studies and usefulness of PMSc® in 3D morpho-volumetric measurements. / Dissertation (MSc (Zoology))--University of Pretoria, 2014. / Zoology and Entomology / MSc (Zoology) / Unrestricted

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[pt] O cabelo humano pode ser utilizado para verificação do estado de saúde do indivíduo. Para tal, são quantificados os elementos essenciais e tóxicos por espectrometria de massa com plasma indutivamente acoplado (ICP-MS) em uma amostra de cabelo e os resultados comparados com valores de referência. Com essas informações, o médico pode avaliar se há algum tipo de desequilíbrio mineral ou intoxicação. O cachorro pode ser um bom bioindicador de exposição ambiental já que ele compartilha do mesmo ambiente que o ser humano, bebe a mesma água e, em alguns casos, come o mesmo alimento. O pelo canino tem quase a mesma composição que o cabelo humano, variando somente sua taxa de crescimento. Neste trabalho, desenvolveu-se um método para determinação dos elementos essenciais e tóxicos em pelo canino por ICP-MS. Para tanto, foram otimizados os procedimentos de lavagem do pelo, a massa mínima de amostra para garantir a homogeneidade, e o procedimento de decomposição. Verificou-se que as melhores condições foram: lavagem com água, massa de amostra entre 100 e 300 mg, e decomposição com HNO3 e H2O2. Os resultados foram comparados com os obtidos por espectrometria de emissão óptica com plasma indutivamente acoplado (ICP OES), técnica já bem estabelecida. O método mostrou-se versátil devido a sua capacidade analítica multielementar, com sensibilidade e acurácia adequadas (os valores de recuperação em relação aos valores certificados ficaram entre 80 e 120 por cento) para a amostra de referência. O trabalho apontou a necessidade de novos estudos para estabelecer valores de referência do mineralograma para cães, assim como avaliar a diferença entre raças. / [en] Human hair can be used to check the individual s health status. For this purpose, the essential and toxic elements are quantified in the inductively coupled plasma mass spectrometer (ICP-MS) and compared to a reference table to identify any possible mineral imbalance or intoxication. The dog can be a good bioindicator of environmental exposure since it shares the same environment as the human being, drink the same water and in some cases eat the same food. Canine fur has almost the same composition as human hair, varying only its growth. In this work, a method was developed to determine the essential and toxic elements in canine by ICP-MS. For that, the hair washing procedures were optimized, the minimal sample mass to ensure homogeneity, and the decomposition procedure. It was found that the best conditions were: washing with water, sample mass between 150 and 300 mg, and decomposition with HNO3 and H2O2. The results were compared with those obtained by optical emission spectrometry with inductively coupled plasma (ICP OES), a well-established technique. The method proved to be versatile due to its multi-element analytical capacity, with adequate sensitivity and accuracy (the recovery values in relation to the certified values were between 80 and 120 per cent) for the reference sample. The work indicated the necessity for further studies to established reference values of the mineralogram for dogs, as well evaluated the difference between races.

Assessment of trophic interactions between increasing populations of New Zealand fur seals (Arctocephalus forsteri) and fisheries in southern Australia is limited due to a lack of species specific demographic data and an understanding of the factors influencing population growth. To establish species specific demographic parameters a cross-sectional sample of New Zealand fur seal females (330) and males (100) were caught and individually-marked on Kangaroo Island, South Australia between 2000 and 2003. The seals were aged through examination of a postcanine tooth, which was removed from each animal to investigate age-specific life-history parameters. Annual formation of cementum layers was confirmed and accuracy in age estimation was determined by examination of teeth removed from individuals of known-age. Indirect methods of assessing reproductive maturity based on mammary teat characteristics indicated that females first gave birth between 4-8 years of age, with an average age at reproductive maturity of 5 years. Among reproductively mature females, age-specific reproductive rates increased rapidly between 4-7 years of age, reaching maximum rates of 70-81% between 8-13 years, and gradually decreased in older females. No females older than 22 years were recorded to pup. Age of first territory tenure in males ranged from 8-10 years. The oldest female and male were 25 and 19 years old, respectively. Post-weaning growth in females was monophasic, characterised by high growth rates in length and mass during the juvenile growth stage, followed by a gradual decline in growth rates after reproductive maturity. In contrast, growth in males was biphasic and displayed a secondary growth spurt in both length and mass, which coincided with sexual and social maturation, followed by a rapid decline in growth rates. Age-specific survival rates were high (0.823-0.953) among prime-age females (8-13 yrs of age) and declined in older females. Relative change in annual pup production was strongly correlated with reproductive rates of prime-age females and adult female survival between breeding seasons.

Les facteurs de transcription NikR et FUR sont impliqués dans l'homéostasie des métaux. Les propriétés de métallation et de liaison à l'ADN de NikR d'E.coli (Ec) et d'H.pylori (Hp) ont été comparées. EcNikR, protéine tétramérique, lie 8 nickels par sous unité dont un nickel dans un site dit de haute affinité ayant un Kd submicromolaire. Les autres sites métalliques, de plus basse affinité, sont impliqués dans le processus l'agrégation Ni- et pH-dépendant. HpNikR, partage des propriétés de métallation avec EcNikR, dont la liaison possible de différents métaux dans son site de haute affinité (Cu(II), Ni(II), Co(II)). Les 4 sites de haute affinité de cette protéine semblent égaux 2 à 2. La métallation des premiers sites faciliterait une « fermeture de la protéine » permettant la métallation des deux derniers sites. La métallation de ces 4 sites semble suffire pour qu'HpNikR puisse lier l'ADN.<br />Cependant cette liaison serait améliorée en présence d'un métal stabilisateur dans d'autres sites. Selon les séquences opératrices, le métal et la technique employée, l‘affinité d'HpNikR métallée pour l'ADN varie. En présence d'un excès de Ni(II), HpNikR se lie à pureA et à pnixA avec un Kd de 2,5 et 1,7 nM mais se lie faiblement à pnikR et pexbB. En présence d'un excès de Ni(II) puis de Mn(II), HpNikR se lie à pnikR et à pexbB avec un Kd de 38 et 24 nM. FUR d'H.pylori (HpFUR), un métallorégulateur Fe dépendant, a aussi été caractérisé dans le but d'étudier sa co-régulation avec HpNikR de la région intergénique nikR-exbB. HpFUR est dimérique et contient au moins deux sites métalliques : un site structural et un site de régulation permettant l'activation pour la liaison à l'ADN.

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.

La régulation de l’homéostasie du fer est cruciale chez les bactéries. Chez Salmonella, l’expression des gènes d’acquisition et du métabolisme du fer au moment approprié est importante pour sa survie et sa virulence. Cette régulation est effectuée par la protéine Fur et les petits ARN non codants RfrA et RfrB. Le rôle de ces régulateurs est d’assurer que le niveau de fer soit assez élevé pour la survie et le métabolisme de Salmonella, et assez faible pour éviter l’effet toxique du fer en présence d’oxygène. Les connaissances concernant le rôle de ces régulateurs ont été principalement obtenues par des études chez S. Typhimurium, un sérovar généraliste causant une gastro-entérite chez les humains. Très peu d’informations sont connues sur le rôle de ces régulateurs chez S. Typhi, un sérovar humain-spécifique responsable de la fièvre typhoïde. Le but de cette étude était de déterminer les rôles de Fur, RfrA et RfrB dans l’homéostasie du fer et la virulence de Salmonella, et de démontrer qu’ils ont une implication distincte chez les sérovars Typhi et Typhimurium. Premièrement, Fur, RfrA et RfrB régulent l’homéostasie du fer de Salmonella. Les résultats de cette étude ont démontré que Fur est requis pour la résistance au stress oxydatif et pour une croissance optimale dans différentes conditions in vitro. La sensibilité du mutant fur est due à l’expression des petits ARN RfrA et RfrB, et cette sensibilité est beaucoup plus importante chez S. Typhi que chez S. Typhimurium. Également, Fur inhibe la transcription des gènes codant pour les sidérophores en conditions riches en fer, tandis que les petits ARN RfrA et RfrB semblent être importants pour la production d’entérobactine et de salmochélines chez S. Typhi lors de conditions pauvres en fer. Ensuite, ces régulateurs affectent la virulence de Salmonella. Fur est important pour la motilité de Salmonella, particulièrement chez S. Typhi. Fur est nécessaire pour l’invasion des deux sérovars dans les cellules épithéliales, et pour l’entrée et la survie de S. Typhi dans les macrophages. Chez S. Typhimurium, Fur ne semble pas impliqué dans l’interaction avec les macrophages. De plus, les petits ARN RfrA et RfrB sont importants pour la multiplication intracellulaire de Salmonella dans les macrophages pour les deux sérovars. Finalement, la protéine Fur et les petits ARN RfrA et RfrB régulent l’expression de l’opéron fimbriaire tcf, absent du génome de S. Typhimurium. Un site de liaison putatif de la protéine Fur a été identifié dans la région promotrice de tcfA chez S. Typhi, mais une régulation directe n’a pas été confirmée. L’expression de tcf est induite par le fer et par Fur, et est inhibée par les petits ARN RfrA et RfrB. Ainsi, ces régulateurs affectent des gènes de virulence qui sont retrouvés spécifiquement chez S. Typhi. En somme, ce projet a permis de démontrer que les régulateurs de l’homéostasie du fer de Salmonella peuvent affecter la résistance de cette bactérie pathogène à différents stress, notamment le stress oxydatif, la croissance en conditions de carence en fer ainsi que la virulence. Ces régulateurs jouent un rôle distinct chez les sérovars Typhi et Typhimurium. / Regulation of iron homeostasis is crucial for bacteria. For Salmonella, proper timing of the expression of iron acquisition and metabolism genes is important for survival and virulence. This regulation is mediated by the protein Fur and the small non-coding RNAs (sRNAs) RfrA and RfrB. The role of these regulators is to assure that the iron level is high enough for survival and metabolism of Salmonella, and low enough to avoid the toxic effect of iron in the presence of oxygen. Thus far, information on the role of these regulators was principally obtained by studying S. Typhimurium, a generalist serovar causing gastro-enteritis in humans. Very little is known about the role of these regulators in S. Typhi, a human-specific serovar which causes typhoid fever. The goal of this study was to determine the roles of Fur, RfrA and RfrB in iron homeostasis and virulence of Salmonella, and to determine if they have a distinct implication in the serovars Typhimurium and Typhi. First, Fur, RfrA and RfrB regulate iron homeostasis in Salmonella. The results of this study have shown that Fur is required for resistance to oxidative stress and for optimal growth in different in vitro conditions. The sensitivity of the fur mutant is due to the expression of the sRNAs RfrA and RfrB, and this sensitivity is worse in S. Typhi than in S. Typhimurium. Also, Fur represses the transcription of the genes encoding siderophores in high-iron conditions, and the sRNAs RfrA and RfrB are required for enterobactin and salmochelins production in S. Typhi in low-iron conditions. Secondly, these regulators affect the virulence of Salmonella. Fur is important for the motility of Salmonella, especially in S. Typhi. Fur is required for the invasion of both serovars in epithelial cells, and for the uptake and survival of S. Typhi in macrophages. In S. Typhimurium, Fur is not required for the interaction with macrophages. Moreover, the sRNAs RfrA and RfrB are important for the intracellular multiplication of Salmonella within macrophages for both serovars. Finally, the Fur protein and the sRNAs RfrA and RfrB regulate the expression of the tcf fimbrial operon, absent from the genome of S. Typhimurium. A putative Fur binding site was identified in the tcfA promoter region of S. Typhi, but direct regulation has not been confirmed. tcf expression is activated by iron and Fur, and is inhibited by the sRNAs RfrA and RfrB. Therefore, these regulators affect virulence genes that are found specifically in S. Typhi. To conclude, this project demonstrates that the iron homeostasis regulators of Salmonella can affect the bacterial resistance to different stresses, espacially oxidative stress, the growth in iron-limiting conditions and virulence. These regulators have a distinct role in the serovars Typhi and Typhimurium.