Developmental and molecular aberrations associated with deterioration of oocytes during FSH-receptor deficiency

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

The effect of nitrate and phosphate availability on Emiliania huxleyi(NZEH) physiology under different CO2 scenarios

Growth and calcification of the marine coccolithophorid Emiliania huxleyi is affected by ocean acidification and macronutrients limitation and its response varies between strains. Here we investigated the physiological performance of a highly calcified E. huxleyi strain, NZEH, in a multiparametric experiment. Cells were exposed to different CO2 levels (ranging from 250 to 1314 µatm) under three nutrient conditions [nutrient replete (R), nitrate limited (-N), and phosphate limited (-P)]. We focused on calcite and organic carbon quotas and on nitrate and phosphate utilization by analyzing the activity of nitrate reductase (NRase) and alkaline phosphatase (APase), respectively. Particulate inorganic (PIC) and organic (POC) carbon quotas increased with increasing CO2 under R conditions but a different pattern was observed under nutrient limitation. The PIC:POC ratio decreased with increasing CO2 in nutrient limited cultures. Coccolith length increased with CO2 under all nutrient conditions but the coccosphere volume varied depending on the nutrient treatment. Maximum APase activity was found at 561 ?atm of CO2 (pH 7.92) in -P cultures and in R conditions, NRase activity increased linearly with CO2. These results suggest that E. huxleyi's competitive ability for nutrient uptake might be altered in future high-CO2 oceans. The combined dataset will be useful in model parameterizations of the carbon cycle and ocean acidification.

Seawater carbonate chemistry, physiology and skeletal mineralogy of coralline alga Corallina elongata in a laboratory experiment

Marine organisms inhabiting environments where pCO2/pH varies naturally are suggested to be relatively resilient to future ocean acidification. To test this hypothesis, the effect of elevated pCO2 was investigated in the articulated coralline red alga Corallina elongata from an intertidal rock pool on the north coast of Brittany (France), where pCO2 naturally varied daily between 70 and 1000 µatm. Metabolism was measured on algae in the laboratory after they had been grown for 3 weeks at pCO2 concentrations of 380, 550, 750 and 1000 µatm. Net and gross primary production, respiration and calcification rates were assessed by measurements of oxygen and total alkalinity fluxes using incubation chambers in the light and dark. Calcite mol % Mg/Ca (mMg/Ca) was analysed in the tips, branches and basal parts of the fronds, as well as in new skeletal structures produced by the algae in the different pCO2 treatments. Respiration, gross primary production and calcification in light and dark were not significantly affected by increased pCO2. Algae grown under elevated pCO2 (550, 750 and 1000 µatm) formed fewer new structures and produced calcite with a lower mMg/Ca ratio relative to those grown under 380 µatm. This study supports the assumption that C. elongata from a tidal pool, where pCO2 fluctuates over diel and seasonal cycles, is relatively robust to elevated pCO2 compared to other recently investigated coralline algae.

Ultrasound imaging in teaching cardiac physiology

This laboratory session provides hands-on experience for students to visualize the beating human heart with ultrasound imaging. Simple views are obtained from which students can directly measure important cardiac dimensions in systole and diastole. This allows students to derive, from first principles, important measures of cardiac function, such as stroke volume, ejection fraction, and cardiac output. By repeating the measurements from a subject after a brief exercise period, an increase in stroke volume and ejection fraction are easily demonstrable, potentially with or without an increase in left ventricular end-diastolic volume (which indicates preload). Thus, factors that affect cardiac performance can readily be discussed. This activity may be performed as a practical demonstration and visualized using an overhead projector or networked computers, concentrating on using the ultrasound images to teach basic physiological principles. This has proved to be highly popular with students, who reported a significant improvement in their understanding of Frank-Starling's law of the heart with ultrasound imaging.

Molecular cloning and expression of leptin in gray and harbor seal blubber, bone marrow, and lung and its potential role in marine mammal respiratory physiology

Leptin is a multifunctional hormone, produced predominantly in adipocytes. It regulates energy balance through its impact on appetite and fat metabolism, and its concentration indicates the size of body fat reserves. Leptin also plays a vital role in stretch-induced surfactant production during alveolar development in the fetus. The structure, expression pattern, and role of leptin have not previously been explored in marine mammals. Phocid seals undergo cyclical changes in body composition as a result of prolonged fasting and intensive foraging bouts and experience rapid, dramatic, and repeated changes in lung volume during diving. Here, we report the tissue-specific expression pattern of leptin in these animals. This is the first demonstration of leptin expression in the lung tissue of a mature mammal, in addition to its expression in the blubber and bone marrow, in common with other animals. We propose a role for leptin in seal pulmonary surfactant production, in addition to its likely role in long-term energy balance. We identify substitutions in the phocine leptin sequence in regions normally highly conserved between widely distinct vertebrate groups, and, using a purified seal leptin antiserum, we confirm the presence of the leptin protein in gray seal lung and serum fractions. Finally, we report the substantial inadequacies of using heterologous antibodies to measure leptin in unextracted gray seal serum.

Ecology, physiology and performance in high-rate anaerobic digestion

The design demands on water and sanitation engineers are rapidly changing. The global population is set to rise from 7 billion to 10 billion by 2083. Urbanisation in developing regions is increasing at such a rate that a predicted 56% of the global population will live in an urban setting by 2025. Compounding these problems, the global water and energy crises are impacting the Global North and South alike. High-rate anaerobic digestion offers a low-cost, low-energy treatment alternative to the energy intensive aerobic technologies used today. Widespread implementation however is hindered by the lack of capacity to engineer high-rate anaerobic digestion for the treatment of complex wastes such as sewage. This thesis utilises the Expanded Granular Sludge Bed bioreactor (EGSB) as a model system in which to study the ecology, physiology and performance of high-rate anaerobic digestion of complex wastes. The impacts of a range of engineered parameters including reactor geometry, wastewater type, operating temperature and organic loading rate are systematically investigated using lab-scale EGSB bioreactors. Next generation sequencing of 16S amplicons is utilised as a means of monitoring microbial ecology. Microbial community physiology is monitored by means of specific methanogenic activity testing and a range of physical and chemical methods are applied to assess reactor performance. Finally, the limit state approach is trialled as a method for testing the EGSB and is proposed as a standard method for biotechnology testing enabling improved process control at full-scale. The arising data is assessed both qualitatively and quantitatively. Lab-scale reactor design is demonstrated to significantly influence the spatial distribution of the underlying ecology and community physiology in lab-scale reactors, a vital finding for both researchers and full-scale plant operators responsible for monitoring EGSB reactors. Recurrent trends in the data indicate that hydrogenotrophic methanogenesis dominates in high-rate anaerobic digestion at both full- and lab-scale when subject to engineered or operational stresses including low-temperature and variable feeding regimes. This is of relevance for those seeking to define new directions in fundamental understanding of syntrophic and competitive relations in methanogenic communities and also to design engineers in determining operating parameters for full-scale digesters. The adoption of the limit state approach enabled identification of biological indicators providing early warning of failure under high-solids loading, a vital insight for those currently working empirically towards the development of new biotechnologies at lab-scale.

Studies on bovine oocytes and ovarian tissues cryopreservation : fundamental and applied aspects

Tese de Doutoramento, Ciências Agrárias (Reprodução Animal), 26 de Junho de 2013, Universidade dos Açores.

Effect of environmental variables on buffaloes physiology

The objective of this work was to evaluate the effect of environmental variables on dairy buffaloes physiology in two different places after milking, shaded plus artificial ventilation and another one non-shaded, in Ribeira Valley, São Paulo State, Brazil. Data on the respiratory rate (RR) and the surface temperature (ST) at udder, neckmiddle, forehead, back middle and rump were collected in 12 dairy buffaloes at autumn. In the same way, it were recorded the black globe temperature in the sun (GTS) and in the shade (GTNS), air temperature and wind speed at padronized height of 1.60 meters. All data were collected at 10:30am and 1:30pm. The results showed statistical difference among black globe temperature, wind speed, RR and ST (P<0.01) in two treatments.The results showed the necessity of protection against the solar radiation in the buffaloes, even in periods of warm climates.

A NOVEL NON-LETHAL LAPAROSCOPIC APPROACH TO DETECT INTERSEX (TESTICULAR OOCYTES) IN LARGEMOUTH BASS (MICROPTERUS SALMOIDES) AND SMALLMOUTH BASS (MICROPTERUS DOLOMIEU)

The appearance of testicular oocytes (TO) in wild fish populations has received considerable attention in the scientific literature and public media. Current methods to quantify TO are lethal; instead, a non-lethal alternative was examined. Laparoscopic insertion into the genital pore allowed internal visualization of the gonad and detection of TO by collecting five testis biopsies in smallmouth bass Micropterus dolomieu and largemouth bass Micropterus salmoides. Overall, biopsies quantified similar levels of TO detection and severity to conventional transverse sectioning with less than 10% mortality. Suitability of surgical anesthetics, tricaine methanesulfonate and electronarcosis were examined in laboratory and field applications. Electronarcosis had the added benefit of rapid sex identification and immediate release of female fish with minimal trauma, representing significant benefits when sampling small or compromised populations. Laparoscopy may be useful for monitoring the prevalence and severity of TO in these fish species when lethal sampling is not a desired outcome.

CHARACTERIZATION OF POSTHARVEST PHYSIOLOGY ATTRIBUTES OF SIX COMMERCIALLY GROWN TOMATO VARIETIES IN KENYA

Tomato ( Lycopersicon esculentum Mill) is the leading vegetable in terms of production in Kenya. The Kenyan local market has a wide variety of tomato cultivars with a wide range of morphological and sensorial characteristics. However, information on the nutritional and postharvest quality of these varieties is lacking. The aim of this research was to investigate and identify tomato varieties of superior postharvest quality and recommend them to small and medium scale farmers. In this study, six tomato varieties were grown in a greenhouse and analyzed at three maturity stages (mature green, turning and red ripe). The tomatoes were analyzed at specific days after harvest and storage at room temperature (25o C). Percentage weight loss, color, respiration and ethylene production rates were analyzed to assess the postharvest quality of the tomatoes. The color was measured using a Minolta Chromameter while the respiration rate and ethylene production rates were determined using the static system approach. Color, weight loss, respiration and ethylene production rates were positively affected by storage time when harvested at the three maturity stages. The percentage weight loss of the tomato fruits was higher in the determinate varieties, and at the turning stage of maturity (3.8 %). Minor color changes were observed after storage of the tomatoes harvested at red stage for six days. Both rates of respiration and ethylene production were low, with the respiration rate ranging between 56-10 ml CO2 Kg-1h-1. The Chonto F1 variety had the highest rate of ethylene production (5.4 μL C2H4 Kg-1h-1) on the 4th day of storage after harvest at the red ripe stage. Overall, the indeterminate tomato varieties displayed better postharvest quality that can prolong the fruits shelf life for marketing. In turn, the turning stage of maturity proved to be a better stage to harvest tomatoes as the color development was more uniform.

Physiology and biochemistry of the effect of abiotic stress on the autochthonous CTP4 strain, a candidate microalga for biofuel production in Algarve

Dissertação de Mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2014

Artificial fertilization of oocytes and sperm activation in pacu: effects of the spermatozoa:oocyte ratio, water volume, and in natura semen preservation

Objetivou-se foi avaliar a fertilização artificial e a duração da motilidade espermática em pacus com diferentes doses inseminantes, volumes de água e preservação do sêmen in natura. Foram realizados quatro experimentos para avaliação do efeito de doses inseminantes (7x10³, 7x10(4), 7x10(5), 7x10(6) e 7x10(7) espermatozoides ovócito-1) sobre a fertilização artificial dos ovócitos; do efeito do volume de água (0,5; 15,0; 30,0; 45,0 e 60,0 mL de água mL-1 de ovócitos) com doses inseminantes de 105.481 e 210.963 espermatozoides ovócito-1; do efeito de diluição do sêmen (0,005; 0,05; 0,5 e 5,0 µL de sêmen mL-1 de água) sobre a duração da motilidade espermática; e do efeito do armazenamento a 15 ºC por 9 h sobre a duração da motilidade espermática e o índice de sobrevivência espermática. Os maiores resultados obtidos foram: doses inseminantes entre 7x10³ e 7x10(7) espermatozoides ovócito-1; 15 a 60 mL de água mL-1 de ovócitos; diluição de 0.005 µL sêmen mL-1 de água e 98,65% de sobrevivência espermática até o tempo de preservação de 2h45min36s. A preservação a 15ºC por 9 horas não influencia a duração da motilidade espermática. As maiores taxas de fertilização podem ser observadas no emprego de 0,27 a 270 µL de sêmen mL-1 de ovócitos, com 15 a 60 mL de água para ativação.

Cold Season Physiology of Arctic Plants

The cold season in the Arctic extends over eight to nine months during which ecosystem gas exchange and water balance of arctic plants have been largely unexplored. The overall objective of this thesis was to examine two critical gaps in our knowledge about tundra cold season processes – ecosystem respiration at very low temperatures and water uptake during the winter-spring transition. I determined the temperature response of ecosystem respiration of tundra monoliths down to temperatures as low as can be expected under snow-covered conditions (-15 °C). Temperature responses fit the Arrhenius function well with Q10 values over the range of -15 to 15 °C varying from 6.1 to 4.8. I used deuterium-enriched water (2H2O) as a tracer to evaluate water uptake of evergreen plants at snowmelt when soils are largely frozen. The results revealed that evergreen plants take up water under snow cover, possibly via roots but undoubtedly by foliar uptake.

Effects of cAMP modulators during in vitro prematuration of bovine oocytes on gap junctional communication and embryo development potential.

2016