Ringed seals (Phoca hispida) in the Canadian Arctic are subject to a variety of environmental and anthropogenic stressors that stand to potentially compromise population health and survival. Typically, animals exposed to chronic stressors initiate a stress response resulting in cortisol production, which results in physiological and behavioural changes designed to maintain homeostasis under the influence of the stressor. Cortisol extraction techniques were developed for Ringed seal blubber and fur samples. Blubber cortisol was found to be a reliable indicator of the condition factor ratio of blubber depth to core diameter. Ringed seal blubber cortisol concentration and condition was shown to alter depending on season and age class. The findings of this study are an important first step in developing an understanding of how this ice obligate species has and may respond to environmental stressors and will assist with developing conservation strategies. / October 2016

Thesis (Ph.D.) - University of St Andrews, April 2010.

Thesis (M.S.)--University of North Carolina at Wilmington, 2004. / Includes bibliographical references.

Dissertação(mestrado) - Universidade Federal do Rio Grande, Programa de Pós–Graduação em Oceanografia Biológica, Instituto de Oceanografia, 2005. / Submitted by Cristiane Gomides (cristiane_gomides@hotmail.com) on 2013-10-09T16:00:18Z No. of bitstreams: 1 Denise.pdf: 786595 bytes, checksum: 5f8cea4a7c4ffd5cc4c20c56dd9c2352 (MD5) / Approved for entry into archive by Sabrina Andrade (sabrinabeatriz@ibest.com.br) on 2013-10-17T03:04:44Z (GMT) No. of bitstreams: 1 Denise.pdf: 786595 bytes, checksum: 5f8cea4a7c4ffd5cc4c20c56dd9c2352 (MD5) / Made available in DSpace on 2013-10-17T03:04:44Z (GMT). No. of bitstreams: 1 Denise.pdf: 786595 bytes, checksum: 5f8cea4a7c4ffd5cc4c20c56dd9c2352 (MD5) Previous issue date: 2005 / A camada de gordura das toninhas, Pontoporia blainvillei, foi estudada partir de medidas de espessura e análise da composição de ácidos graxos. Os animais analisados foram capturados acidentalmente pele frota pesqueira de emalhe do Rio Grande (32º 05’S e 52º 08’W), entre 1999 e 2003. Com objetivo de investigar a possibilidade do uso da espessura da camada de gordura na estimativa da condição corporal nesta espécie, foram tomadas medidas de espessura em 116 indivíduos. As medidas de espessura foram obtidas nas regiões dorsal, ventral e lateral de uma mesma circunferência do corpo, localizada entre as nadadeiras peitoral e dorsal. A maturidade sexual dos animais foi determinada a partir do exame macroscópico e histológico das gônadas. A composição e a estratificação de ácidos graxos (AG) foi estudada a partir de amostras da região médio-ventral da camada de gordura de 30 indivíduos, sendo a idade destes estimada através dos grupos de camadas de crescimento dos dentes. As amostras da camada de gordura foram subdivididas em três estratos de profundidade (pele-músculo). Os lipídios de cada estrato foram extraídos utilizando-se solução (2:1) de clorofórmiometanol e os ésteres metílicos de ácidos graxos foram extraídos com solução BF3 – metanol e posteriormente analisados por cromatografia gás-líquida. A identificação de cada ácido graxo foi feita pela comparação dos tempos de retenção das amostras com o de misturas-padrão comercializadas. Os filhotes apresentaram menor espessura de camada de gordura na região dorsal em relação aos maduros e imaturos. Enquanto os machos não apresentaram diferenças entre maduros e imaturos com relação à espessura, fêmeas maduras possuíram maior espessura da camada de gordura dorsal do que as imaturas. Foi detectada variação sazonal a partir da espessura da camada de gordura apenas na região ventral, sendo que as menores espessuras foram registradas no outono e as maiores no inverno. A variação interanual foi evidenciada na espessura dorsal da camada, sendo que os indivíduos capturados em 2002 apresentaram melhor condição corporal do que os indivíduos capturados em 2000 e 2001, o que possivelmente foi resultante de mudanças nas condições ambientais e na disponibilidade de presas. Um total de 27 ácidos graxos foram identificados, sendo que o 16:1n-7 e o 18:1n-9 estiveram presentes em maiores concentrações (em média 37,14% e 19,70%, respectivamente). A maioria dos AG apresentou gradiente contínuo através dos estratos, aumentando ou diminuindo de concentração do sentido externo (adjacente à pele) para o interno (adjacente ao músculo). Os AG saturados e monoinsaturados de cadeias mais curtas (12:0, 18:0, 14:1n-5 e 15:1) apresentaram os maiores graus de estratificação, aumentando de concentração no sentido do estrato externo. Já os AG poliinsaturados (PUFAs) 20:3, 20:5n-3, 22:6n-3, 20:4n-6 e 22:2 e o monoenóico 24:1n-9 apresentaram maior estratificação no sentido do estrato interno. A composição e a estratificação dos AG também foram diferentes entre as classes etárias e entre os sexos. O total de PUFAs no estrato interno aumentou com a idade e esta relação foi mais forte em machos. O total de AG saturados presentes no estrato externo diminuiu com a idade, sendo esta relação mais forte em fêmeas. Não foram encontradas correlações entre a idade e a razão de omega-3/ omega-6 para os estratos externo ou interno. Entre os adultos, os machos apresentaram maior grau de estratificação do que fêmeas. O presente estudo mostra a possibilidade de utilizar-se a espessura da camada de gordura na região dorsal como um indicador da condição corporal em toninhas e evidencia variabilidade na composição de AG dos indivíduos, dando aporte para futuras investigações sobre análises de dieta ou diferenciação de estoques populacionais nesta espécie através da composição de AG. / In order to investigate the usefullness of blubber thickness to estimate the body fat condition in Pontoporia blainvillei, axillary girth measurements were taken from dorsal, ventral and lateral regions, between the pectoral flipper and the dorsal fin. Blubber measurements were obtained for 116 franciscana dolphins, accidentally caught by the coastal artisanal fishery from Rio Grande (32º 05’S e 52º 08’W), between 1999 e 2003. Sexual maturity was determined from macroscopic and histological analysis of gonads. Dorsal blubber thickness (DBT) had the highest variation among the 3 measurements taken, and was considered the best measurement assess body fat condition in this species. DBT was lowest for pups and highest for mature females. Immature and mature males did not differ in DBT measurements. Dorsal blubber thickness also exhibited inter-annual variation such that 2002 individuals had thicker layers and thus better body condition that those accidentally caught in 2000 and 2001. Ventral blubber thickness (VBT) did not vary as much as DBT but seemed to be affected by seasonality. VBT was lowest in the fall and highest in the winter. Interannual variation in blubber thickness may be the result of changing environment conditions and/or prey composition, condition and availability. This study showed the potential use of blubber thickness as a proxy to estimate body fat condition in franciscana dolphins. Additionally, the fatty acid composition and stratrification of the blubber layer (skin-muscle) at the middle-ventral region was studied in 30 franciscanas. The lipids were extracted using chloroform-methanol and fatty acids methyl esters were obtained using BF3-methanol. Analyses were performed by gas-liquid chromatography. Fatty acids were identified by comparison of the retention time against commercial standards. Age determination estimated by counting GLGs (Growth Layer Groups) from teeth. Twenty-seven fatty acids were quantified (27/37), of which 16:1n-7 (37,14%) and 18:1n-9 (19,70%) exhibited the highest concentrations. Stratification of fatty acid concentration across the blubber layer was found where a continuous gradient of FA concentrations was evident either increasing or decreasing from the inner layer towards the skin. The shorter-chain saturated and monounsaturated fatty acids (12:0, 18:0, 15:1 and 14:1n-5) exhibited the highest relative amounts in the outer layer whereas polyunsaturated fatty acids (PUFA) 20:3, 20:5n-3, 22:6n-3, 20:4n-6 and 22:2 and the monounsaturated fatty acid 24:1n-9 exhibited the highest relative amounts in the inner blubber layer. The array of fatty acids identified differed among age categories and between females and males. Total PUFA in the inner layer increased with the age and this relationship was stronger for male franciscanas. On the other hand, total saturated fatty acids found in the outer layer decreased with the age an this relationship was stronger for females. There was no correlation between the ratio omega-3/ omega-6 and the age of the individuals for either the outer or inner blubber layer. The strongest degree of stratification was exhibited by adult males. This study describes for the first time the existence of stratification and the fatty acid composition of the blubber layer of franciscana dolphins from FMA IIIs.

This thesis examines fatty acid (FA) and polychlorinated biphenyl (PCB) dynamics in a marine top predator, the grey seal (Halichoerus grypus,) and their transfer during lactation from mother to offspring. It examines regional and annual variations in FA composition and PCB loads, and also how the physical and chemical characteristics of these molecules (e.g. their polarity and size) can affect the rates of accumulation, mobilization and transfer of specific FAs or PCBs. Two UK grey seal colonies (North Rona (NR) and Isle of May (IOM) were studied during three consecutive years (1996-1998 and 2004-2006). Lactating grey seals and their pups were repeatedly captured during the lactation period and sampled for blubber, serum and milk and analysed for FAs and PCBs. Overall, the two colonies were clearly distinguished from each other, suggesting that the main prey species had different FA composition, and possibly that the seals from these colonies had different diets . These differences are probably a direct consequence of differences in prey community structure in the two regions where seals from these two colonies are thought to feed. Within each colony, annual differences could be detected between some years but not between others. During 1996-98, IOM seals showed a clear change in their FA profiles while NR seals did not. In contrast, during 2004-2006 NR seals showed a clear change while IOM seals did not. The changes observed in IOM during 1996-1998 are consistent with the large-scale regime shift that occurred in the North Sea during the 1990‟s. The relative proportions of each FA that were mobilized from blubber and transferred to the milk during lactation were very similar between colonies, and could be explained to a large degree by their physico-chemical properties. For a given carbon chain length the mobilization increased with increasing number of double bonds; and for a given number of double bonds the mobilization decrease with increasing carbon chain length. However, the mobilization also appeared to be influenced by the specific nutritional requirements of the growing pups. For instance, FAs that are considered essential for pup development or efficient energy storage (e.g. saturated FAs) were more highly mobilised than expected. This selectivity was also reflected in the FA composition of the different body compartments (maternal blubber and milk, pup blubber) that persisted throughout lactation. These changes were also similar between the colonies. Colonies could also be clearly distinguished by their blubber PCB profiles. IOM seals had higher total concentrations on average than NR seals (1327.9 vs. 680.2 ng/g lipid in 2005 and 1199.7 vs. 819.0 ng/g lipid in 2006). IOM seals also had higher total amounts in both years (79.2 vs. 38.0 mg in 2005 and 61.7 vs. 53.4 mg in 2006). One of the main differences between colonies was that females from IOM had higher concentrations of highly chlorinated congeners than NR seals. PCB concentrations in blubber increased towards the end of lactation. Serum and milk PCB concentrations also increased rapidly, especially for the highly chlorinated congeners. These results were consistent with other studies showing the increase in concentrations as a result of lipid loss. Serum concentrations stayed constant during the first part of lactation and increased at late lactation. This was also observed in milk PCB concentrations. The changes in the PCB profiles in the three body compartments were very similar between colonies. However IOM seals always had higher total concentrations of PCBs in all of the body compartments. The concentrations of individual congeners relative to PCB-153 showed that blubber contained higher proportions of the highly chlorinated PCBs relative to other tissues. There were no clear changes in these proportions in blubber during lactation, but the relative proportions of highly chlorinated PCB In serum and milk increased throughout lactation while the less chlorinated PCBs stayed constant. The highly chlorinated PCBs were found in lower concentration in the milk compared to the less chlorinated compounds suggesting a selective release from blubber to blood and a selective transfer of PCBs to the milk.

Orientador: Carolina Pacheco Bertozzi / Abstract: The Franciscana dolphin (Pontoporia blanvillei) is a small odontocete impacted by anthropic activities (bycatch, habitat degradation). The hormonal measurement in biological matrices (feces, urine, blubber, milk, saliva, blow) has been increasingly used in aquatic mammals for certain reproductive parameters. The present study validated the method of extraction and measurement of testosterone and progesterone in blubber kept in two different temperatures (-20°C and -196°C), besides correlating the hormone concentration with gender, sexual maturity, age, stage of development, body length and occurrence of pregnancy. The carcasses were collected from stranded and bycatch dolphins of the coast of São Paulo. Blubber samples were collected and frozen at -20°C (n=101) and -196°C (liquid nitrogen) (n=28). The extraction was performed with a method described in literature and the hormonal analyses were done by enzyme immunoassay for the two hormones. An analysis was performed on two units of hormone concentration (ng/g and ng/mg). The inter-assay coefficient of variation was 6.89% for progesterone and 1.67% for testosterone, the intra-assay coefficient of variation was 7.82% for progesterone and 3.29% for testosterone. The progesterone assay presented parallelism F1.5= 0.87, p= 0.45 and testosterone presented F1.5= 3.79, p= 0.11. The mean extraction efficiency was 66% for progesterone and 70% for testosterone. The method of extraction and hormonal analysis was feasible for Pontoporia ... (Complete abstract click electronic access below) / Resumo: A toninha (Pontoporia blanvillei) é um pequeno odontoceto impactado pelas atividades antrópicas (capturas acidentais, degradação de habitat). A dosagem hormonal em diferentes matrizes (fezes, urina, tecido adiposo, leite, saliva, borrifo) tem sido cada vez mais utilizada em mamíferos aquáticos para determinar parâmetros reprodutivos. O objetivo deste estudo foi validar o método de extração e mensuração de testosterona e progesterona em camada de gordura subcutânea de carcaças de toninha armazenadas em duas temperaturas (-20°C e -196ºC), além de correlacionar as dosagens com parâmetros de sexo, maturidade gonadal, idade, estágio de desenvolvimento, comprimento total e ocorrência de prenhez. As amostras foram colhidas de carcaças provenientes de encalhe e captura acidental do litoral de São Paulo. Foram colhidos fragmentos de gordura, armazenados em freezer a -20°C (n=101) e em nitrogênio líquido (-196ºC) (n=28). A extração hormonal foi realizada com modificações a partir do método descrito em literatura e a dosagem foi feita por enzimaimunoensaio para os dois hormônios. Os resultados das análises estatísticas foram expressos em duas unidades de concentração de hormônio (ng/g e ng/mg). O coeficiente de variação inter-ensaio foi de 6,89% para progesterona e 1,67% para testosterona, o coeficiente de variação intra-ensaio foi de 7,82% para progesterona e 3,29% para testosterona. Os resultados do paralelismo para o ensaio de progesterona foi F1,5 =0,87 e p=0,45 e para testosterona ... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

The conservation of marine mammals relies on the knowledge of species ecology in order to assess the impacts of anthropogenic activites and make appropriate species management decisions. Blubber biopsies are a relatively non-invasive sampling protocol to provide ecological information; two particular uses are for dietary analysis via fatty acid signature analysis (FASA) and investigating the uptake of organochlorine (OC) and polychlorinated biphenyl (PCB) environmental contaminants. Blubber composition and structure may vary according to body site and depth due to its dynamic functioning. This may result in the vertical stratification and heterogenous distribution of blubber FAs, OCs and PCBs between body sites, giving variable interpretations of diet and contaminant levels depending on biopsy sample site and depth. The aim of this thesis is to determine the FA distribution of blubber from two body sample sites (dorsal pelvic and ventral thoracic) currently used for FASA of the New Zealand sea lion, Phocarctos hookeri (NZSL) and to determine the level of OCs and PCBs in the blubber of healthy NZSLs for comparison to diseased NZSLs recorded in the 1997/98 epidemic. Blubber samples were collected from 29 by-caught NZSLs incidentally captured by the squid fishery around the Auckland Islands (50º42’S, 166º5’E) during the years 2005 to 2007 (not all NZSLs were able to be analysed for each chapter). Full blubber cores from both sample sites were collected from 18 NZSLs. Both sites showed a relative homogeneity of FA profiles, indicating the similar deposition and mobilisation of FAs at the two sample sites. To determine if FA stratification occurred, full blubber cores from both sample sites of 20 NZSLs were divided into inner and outer halves. Both sites displayed the same pattern of vertical stratification or biochemical layering of FAs between the two divisions, indicating that stratification of FAs occurs in the blubber of the NZSL. A range of OCs and PCBs were then determined in full thoracic blubber cores of seven NZSLs. The levels were higher than those previously recorded in NZSLs affected by disease during a 1997/98 epidemic. The SPCB in NZSLs was 0.034-0.192 mg/kg lipids, below the suggested threshold of 17 mg/kg for adverse health affects in marine mammals. From the results of this study I can support current blubber biopsy sampling techniques for FASA in NZSL. Obtaining full blubber cores from either the thoracic or pelvic sample site will give a comparable interpretation of diet. The low levels of blubber OCs and PCBs suggest a minor role of contaminants acting as a possible causative agent toward disease outbreaks in the NZSL. This research provides important information for developing correct and consistent blubber sampling techniques for NZSL and other pinniped species. This will ensure more accurate interpretations of ecological information obtained from blubber biopsies and therefore improved species management and conservation decisions that may be based on such research.

No / Shallow oval bowls used on the Baltic coast in the Mesolithic have been suggested as oil lamps, burning animal fat. Here researchers confirm the use of four coastal examples as lamps burning blubber-the fat of marine animals, while an inland example burned fat from terrestrial mammals or freshwater aquatics-perhaps eels. The authors use a combination of lipid biomarker and bulk and single-compound carbon isotope analysis to indicate the origin of the residues in these vessels.