Genetics of berry colour and anthocyanin content variation in grapevine (Vitis vinifera L.subsp. vinifera)

Dissertation presented in fulfillment of the requirements for the Degree of Doctor of Philosophy in Biology (Molecular Genetics) at the Instituto de Tecnologia Química e Biológica da Universidade Nova de Lisboa

A systems biology framework for pathway level culture media engineering: pplication to Pichia pastoris cultures

Dissertação para obtenção do Grau de Doutor em Engenharia Química e Bioquímica

Biology of Triatominae (Reduviidae Hemiptera) from North of Formosa County (Goiás-Brazil) I. Length of life cycle of Triatoma sordida (Stal. 1859)

In the present paper the life cycle of Triatoma sordida was studied. The mean length from egg to adult was 213 days. The mean length in days from each stage was: 24.3 (± 1.30) for the first. 32.8 (± 1.45) (2nd), 36.1 (± 1.50) (3rd), 44.6 (± 1.85) (4th) and 52.0 (± 1.92) (5th). The mean egg incubation períod was 23.2 (± 1.40). Overall mortality was 18.8% and egg viability was 82.5%.

Biology of Triatominae (Reduviidae, Hemiptera) from north of formosa county (Goiás - Brazil) II. Length of life cycle of Rhodnius Neglectus lent, 1954

In the present paper the life cycle of Rhodnius neglectus was studied. The mean length from egg to adult was 119 days. The mean length in day from each stage was: 17.4 (± 1.15) for first, 18.2 (± 1.28) for second, fourth and 29.8 (± 1.46) for fifth.

Biology of triatominae (Reduviidade, Hemiptera) from north of Formosa county (Goiás - Brazil). III: length of life cycle of Psammolestes tertius Lent and Jurberg 1965

In the present work the life cycle of Psammolestes tertius was studied. The mean length, in days, fromeach stage was: 26.3 (± 1.7) (1st), 28.6 (± 1.8) (2nd), 28.4 (± 1.8) (3rd), 32.2 (± 1.9) (4th) and 33.5 (± 5.8) (5th). The mean egg incubation period was 15.7 days (± 1.7). Overall mortality was 48.9% and egg viability was 65.7%.

FSR Quorum Sensing: Role in Enterococcus faecalis Biology & Host Infection

Dissertation presented to obtain the Ph.D degree in Biology.

Comparative biology of the two sister species of Triatominae (Hemiptera: Reduviidae)

INTRODUCTION: Triatoma pseudomaculata and T. wygodzinskyi (Hemiptera: Reduviidae: Triatominae) are two Brazilian vectors of Chagas disease. The first is an arboricolous species in sylvatic environment and considered a vector of T. cruzi in peridomestic structures; the second, a rupicolous species in the wild environment of no epidemiological importance. In order to test the assumption that sister species share biological traits, comparative studies of their development cycle and blood ingestion were conducted. METHODS: Eggs laid by five field females of each species were randomly selected. The nymphs were observed daily and fed on mice weekly. The time required to pass through the different stages to adulthood was recorded in days. The triatomines were weighed individually before and after feeding. The mortality rate according to each nymphal stage was calculated. RESULTS AND CONCLUSIONS: Analysis of the results shows that they display only minor biological differences even though they exhibit a distinct ecology. This suggests that the biological traits are important criteria to determine the relationship between species.

Biology of Triatoma carcavalloi Jurberg, Rocha & Lent, 1998 under laboratory conditions

Introduction Triatoma carcavalloi is a wild species that is found in sympatry with Triatoma rubrovaria and Triatoma circummaculata, which are vectors of Trypanosoma cruzi currently found in rural areas of Rio Grande do Sul, Brazil. Methods Fertility was assessed and to determine the incubation period, the eggs were observed until hatching. The first meal was offered to 1st stage nymphs. The intermolt period was also determined. The number of blood meals was quantified at each nymphal stage and the resistance to fasting as the period between ecdysis and death. Mortality was assessed and longevity was determined by recording the time that elapsed from molting to the adult stage and until death. The developmental cycle was assessed by recording the length in days of each stage from molting to adult hood. Results The average incubation period was 22.7 days. The average first meal occurred 3.1 days after hatching. The 5th stage nymph to adult intermolting period was the longest at 193.4 days. The average number of feedings during nymphal development was 13.4. The resistance to fasting assay indicated that the 3rd, 4th and 5th stage nymphs presented higher resistance than did adults. The highest mortality rate was observed in the 3rd stage nymphs (22.2%). The average length of adult survival was 25.6 weeks, and the average total life cycle lasted 503.4 days. Conclusions This study is the first report on the biology of T. carcavalloi that fed on mice. The presented findings expand the bionomic knowledge of these species.

Ecoepidemiology and biology of Eratyrus mucronatus Stål, 1859 (Hemiptera: Reduviidae: Triatominae), a sylvatic vector of Chagas disease in the Brazilian Amazon

Introduction Eratyrus mucronatus Stål, 1859 is a wild triatomine vector of Trypanosoma cruzi Chagas, 1909. However, little is known regarding the biology and ecoepidemiology of this triatomine in the Brazilian Amazon. The present study describes the biology of E. mucronatus grown under laboratory conditions and the epidemiological aspects of its natural breeding sites. Methods Five colonies were monitored in the field for 3 years. Temperature and humidity measurements were taken in the mornings and afternoons at the natural breeding sites, and the behavior and distribution of the nymphs and adults were observed in the wild colony. We also monitored the life cycle under controlled laboratory conditions. Results Some factors that were considered decisive for the establishment of these colonies were present at all of the colonies studied in the field. These factors included an active termite nest, a vertebrate for repast, and dry and shaded substrates with temperatures of 24-28°C and with humidity of 80-90%. A generation was developed in 274 days under these microclimatic conditions in the laboratory. Conclusions The climatic variables described in the field indicate that these environmental parameters have a limiting effect on the dispersal and colonization of E. mucronatus to new environments. In addition, the long period of development to adulthood demonstrates that only one generation can develop per year even under the more favorable laboratory conditions.

Development of process control strategies exploiting knowledge from systems biology: application to MDCK suspension cells

Madine Darby Canine Kidney (MDCK) cell lines have been extensively evaluated for their potential as host cells for influenza vaccine production. Recent studies allowed the cultivation of these cells in a fully defined medium and in suspension. However, reaching high cell densities in animal cell cultures still remains a challenge. To address this shortcoming, a combined methodology allied with knowledge from systems biology was reported to study the impact of the cell environment on the flux distribution. An optimization of the medium composition was proposed for both a batch and a continuous system in order to reach higher cell densities. To obtain insight into the metabolic activity of these cells, a detailed metabolic model previously developed by Wahl A. et. al was used. The experimental data of four cultivations of MDCK suspension cells, grown under different conditions and used in this work came from the Max Planck Institute, Magdeburg, Germany. Classical metabolic flux analysis (MFA) was used to estimate the intracellular flux distribution of each cultivation and then combined with partial least squares (PLS) method to establish a link between the estimated metabolic state and the cell environment. The validation of the MFA model was made and its consistency checked. The resulted PLS model explained almost 70% of the variance present in the flux distribution. The medium optimization for the continuous system and for the batch system resulted in higher biomass growth rates than the ones obtained experimentally, 0.034 h-1 and 0.030 h-1, respectively, thus reducing in almost 10 hours the duplication time. Additionally, the optimal medium obtained for the continuous system almost did not consider pyruvate. Overall the proposed methodology seems to be effective and both proposed medium optimizations seem to be promising to reach high cell densities.