The genetics of behaviour and other adaptive traits in nine-spined sticklebacks (Pungitius pungitius)

One of the main goals in current evolutionary biology research is to identify genes behind adaptive phenotypic variations. The advances in genomic technologies have made it possible to identify genetic loci behind these variations, also concerning non-model species. This thesis investigates the genetics of the behaviour and other adaptive traits of the nine-spined stickleback (Pungitius pungitius) through the application of different genetic approaches. Fennoscandian nine-spined stickleback populations express large phenotypical differences especially in behaviour, life –history traits and morphology. However the underlying genetic bases for these phenotypical differences have not been studied in detail. The results of the project will lay the foundation for further genetics studies and provide valuable information for our understanding of the genetics of the adaptive divergence of the nine-spined stickleback. A candidate gene approach was used to develop microsatellite markers situating close to candidate genes for behaviour in the nine-spined stickleback. Altogether 13 markers were developed and these markers were used in the subsequent studies with the anonymous random markers and physiologically important gene markers which are already currently available for nine-spined sticklebacks. It was shown that heterozygosity correlated with behaviour in one of the marine nine-spined stickleback populations but with contrasting effects: correlations with behaviour were negative when using physiological gene markers and positive with random markers. No correlation was found between behavioural markers and behaviour. From the physiological gene markers, a strong correlation was found between osmoregulation-related gene markers and behaviour. These results indicate that both local (physiological) and general (random) effects are important in the shaping of behaviour and that heterozygosity– behaviour correlations are population dependent. In this thesis a second linkage map for nine-spined sticklebacks was constructed. Compared to the earlier nine-spined stickleback linkage map, genomic rearrangements were observed between autosomal (LG7) and sex-determing (LG12) linkage groups. This newly constructed map was used in QTL mapping studies in order to locate genomic regions associated with pelvic structures, behaviour and body size/growth. One major QTL was found for pelvic structures and Pitx1 gene was related to these traits as was predicted from three-spined stickleback studies, but this was in contrast to earlier nine-spined stickleback study. The QTL studies also revealed that behaviour and body size/growth were genetically more complex by having more QTL than pelvic traits. However, in many cases, pelvic structure, body size/growth and behaviour were linked to similar map locations indicating possible pleiotropic effects of genes locating in these QTL regions. Many of the gene related markers resided in the QTL area. In the future, studying these possible candidate genes in depth might reveal the underlying mechanism behind the measured traits.

Glyphosate translocation in hairy fleabane (Conyza bonariensis) biotypes

The objective of this work was to evaluate the translocation of glyphosate in C. bonariensis plants resistant and susceptible to that herbicide. The 14C-glyphosate was mixed with commercial gyhphosate (800 g ha-1) and applied on the center of the adaxial face of a third node leaf, using a micro syringe, and adding 10 µL of a solution with specific activity of 1,400 Bq, 45 days after plant emergence. The concentration of the glyphosate translocated in the plant was evaluated at time intervals of 6, 12, 36 and 72 hours after being applied on the application leaf, stem, roots and leaves. Ten hours after treatment application, the distribution of the product in the application leaf, divided into base, center and apex, was also evaluated by measuring the radiation emitted by 14C-glyphosate in a liquid scintillation spectrometer. Greater glyphosate retention was observed in the resistant biotype leaf, approximately 90% of the total absorbed up to 72 hours. In the susceptible biotype, this value was close to 70% in the same period. Susceptible biotype leaves, stem and roots showed greater concentration of glyphosate, indicating greater translocation efficiency in this biotype. In the resistant biotype, the herbicide accumulated in greater quantity at the apex and center of the application leaf, while in the susceptible biotype greater accumulation was observed at the base and center leaf. Thus, it can be stated that the resistance mechanism is related to the differential translocation of this herbicide in the biotypes.

Control of glyphosate resistant hairy fleabane (Conyza bonariensis) with dicamba and 2,4-D

Auxyn type herbicides such as dicamba and 2,4-D are alternative herbicides that can be used to control glyphosate-resistant hairy fleabane. With the forthcoming possibility of releasing dicamba-resistant and 2,4-D-resistant crops, use of these growth regulator herbicides will likely be an alternative that can be applied to the control of glyphosate resistant hairy fleabane (Conyza bonariensis). The objective of this research was to model the efficacy, through dose-response curves, of glyphosate, 2,4-D, isolated dicamba and glyphosatedicamba combinations to control a brazilian hairy fleabane population resistant to glyphosate. The greenhouse dose-response studies were conducted as a completely randomized experimental design, and the rates used for dose response curve construction were 0, 120, 240, 480, 720 and 960 g a.i. ha-1 for 2,4-D, dicamba and the dicamba combination, with glyphosate at 540 g a.e. ha-1. The rates for glyphosate alone were 0, 180, 360, 540, 720 and 960 g a.e. ha-1. Herbicides were applied when the plants were in a vegetative stage with 10 to 12 leaves and height between 12 and 15 cm. Hairy fleabane had low sensitivity to glyphosate, with poor control even at the 960 g a.e. ha-1 rate. Dicamba and 2,4-D were effective in controlling the studied hairy fleabane. Hairy fleabane responds differently to 2,4-D and dicamba. The combination of glyphosate and dicamba was not antagonistic to hairy fleabane control, and glyphosate may cause an additive effect on the control, despite the population resistance.

Genetic interactions involving major genes at the dwarf locus in egg-type chickens

Three egg-type stocks segregating dwarf (dw) and bantam (dwB) genes in female progeny were produced from the same 18 heterozygous (dwB/dw) sires used to inseminate dams of three different genotypes: normal (dw+), dwarf (dw) and bantam (dwB) dams. The heritability of 8-week body weight estimated from full-sibs of the same phenotype of progeny was 0.40, and that estimated from paternal half-sibs of the same phenotype (dwarf or bantam), and from the same genotype of dam was 0.38. Therefore, maternal and non-additive effects within genotypic classes of dam made little contribution to the genetic variance for 8-week body weight among their progeny. The interaction of sires (S) with genotypes (dw+, dw and dwB) of dam (G) was significant at the 5% level. This indicates that the rankings of the sires within each one of the three genotypes of dam were not the same, probably due to non-additive genetic variation among genotypes of dams. The evidence indicated that in general the genes from individual sires combined differently with each type of dam (G). Those genes which combined well with the genes from normal (dw+) dams combined poorly with both the genes from the dwarf (dw) and the genes from the bantam (dwB) dams. The interaction of sires (S) with phenotypes (dwarf and bantam) of progeny (P) was significant at the 10% level. The results indicated a probable gene x genotype interaction for 8-week weight between genes at the dwarf locus (dw and dwB) and the background genotype (single and/or polygenes). The correlation among paternal half-sibs was influenced more by the S x G than by the S x P interaction, but the effects tended to be cumulative

Synthesis of stocks and phenotypic effects of dwarf and bantam sex-linked major genes in egg-type chickens

The present study describes the production of stocks segregating dwarf (dw), bantam (dwB) and normal (dw+) alleles, as well as the characters, shank length, adult body weight, age at sexual maturity and egg production. Heterozygous K dw+/k dwB sires were mated to normal (dw+) dams to produce stock D6.a, and mated to dwB females to produce stock D6.b. Stock D4.a came from mating F1 heterozygous dwB dw sires to dwarf Leghorns. In a third series of matings, 7/8 Sebright and 1/8 dw-Leghorn dwB dw sires were crossed to three groups of dams of different genotypes. The progeny of the normal (dw+), dwarf (dw), and bantam (dwB) dams were designated as stocks D4.b, D4.c and D4.d, respectively. The dw+ dams were White Leghorn strain cross females. The difference between the rate of laying of normal (69.7%) and their bantam sisters (68.6%) was not statistically significant when the average 32-week body weight of the dw+ sisters was 1,897 g. However, when the 32-week body weight of the normal daughters from the same sires and smaller dams was around 1,646 g, the difference between the rate of laying of the normal (78.1%) and their bantam sisters (75.9%) was significant (P < 0.05). The dwB gene may have a similar but smaller effect on the rate of egg laying than its dwarf allele. The difference between sexual maturity of normal and bantam daughters of either the largest or the smallest dams was not statistically significant, even though the smallest dwB pullets were in average 2.9 days older at first egg. The use of shank length combined with adult body weight allowed a precise discrimination between bantams and dwarfs

Tangential microfiltration of orange juice in bench pilot

The aim of this study was to introduce the tangential microfiltration (TMF) technique on the production of orange juice (TMFJ), and compare it with pasteurised juice (control) as regards chemical composition and sensorial characteristics. We used a TMF pilot equipped with four monotubular ceramic membranes (0.1, 0.2, 0.8 and 1.4mm) arranged in series with a filtering area of 0.005 m² each. Commercial flash-pasteurised orange juice was used as the initial product. Experiments were divided into three parts: a) the characterisation of the TMF pilot; b) optimisation of operational conditions; c) production of the TMFJ. In the second part, membrane with 0.8-mm pores presented best flux followed by those with 1.4-, 0.1-, and 0.2-mm pores. However, to guarantee permeate sterility, we chose the membrane with 0.1-mm pores for TMFJ production. Initially, the orange juice was sieved in order to separate part of the pulp, being subsequently submitted to TMF. A mixture of retentate and pulp was made, and was subsequently pasteurised. We obtained the TMFJ by adding the permeate to the mixture. TMFJ presented soluble solids content (°Brix), pulp, pH, and titrable acidity similar to the initial pasteurised juice (control). Nevertheless, 28% of vitamin C was lost during the TMFJ production. According to the juice taster panel, the control juice presented best sensorial characteristics (greater aroma intensity and fruity flavour) when compared with the TMJF.

Dynamics of gas levels inside packages containing minimally processed Pera orange

The purpose of this study was to evaluate the changes in concentrations of O2 and CO2 inside packages of minimally processed Pera orange. Previously selected oranges that were washed, sanitized, and chilled were peeled using hydrothermal treatment (immersion of fruits in water at 50 °C for 8 minutes). The peeled oranges were then packed in five different plastic packages under passive and active modified atmosphere (5% O2 + 10% CO2 + 85% N2). The fruits were stored at 6 °C and 12 °C. The package headspace gas composition was evaluated for twelve days at 6 °C and nine days at 12 °C. The polypropylene film (32 µm) promoted modified atmosphere similar to that initially injected (5% O2 + 10% CO2 + 85% N2) at 6 °C and 12 °C. With regard to the atmosphere modification system, the injection of a gas mixture anticipated achieving an equilibrium atmosphere inside the packages at 12 °C. At 6 °C, the gas composition inside the packages was kept close to that of the injection, but the equilibrium was not verified.

Quality of 'Delta Valencia' orange grown in semiarid climate and stored under refrigeration after coating with wax

The effects of carnauba-based wax on the quality of 'Delta Valencia' orange produced in Ceará state, Brazil, were studied. The fruits were coated with carnauba-based wax and refrigerated (7 ± 2 ºC and 85 ± 2% R.H.) for 28 days. The quality attribute parameters assessed were weight loss, peel color (brightness, hue angle, and chromaticity), peel moisture, pH, soluble solids (SS), titratable acidity (TA), SS/TA ratio, ascorbic acid, total soluble sugars, reducing sugars, yellow flavonoids, and polyphenols. The results showed that 'Delta Valencia' oranges grown in the dry climate of Ceará state has excellent quality. The coated fruits lost mass at a lower rate than the the control fruits. No significant loss of soluble solids, titratable acidity, pH, and SS/TA ratio was observed, while ascorbic acid, soluble sugars, reducing sugars, yellow flavonoids, and polyphenols increased during storage in both the coated and control fruits. Carnauba-based wax coated fruits showed no signal of dehydration keeping their shiny green peel up to the end of the storage. The use of coating was crucial for the maintenance of visual quality by reducing mass loss, as well as keeping peel moisture.

Study of the enthalpy-entropy mechanism from water sorption of orange seeds (C. sinensis cv. Brazilian) for the use of agro-industrial residues as a possible source of vegetable oil production

Orange seeds are a promising agroindustry-waste which can be implemented in the extraction and production of vegetable oil. The relationship between moisture content and water activity provides useful information for the processing and storage of this waste item. The aim of this study was to determine the mechanism of water sorption enthalpy-entropy of orange seeds (C. sinensis cv. Brazilians) according to the moisture content. Therefore, desorption isotherms were determined at five different temperature (30, 40, 50, 60, and 70 ºC) under a wide range of moisture content (0.005-0.057 kg kg-1 d.b.) and water activity (0.02-0.756). Theoretical and empirical models were used for modeling the desorption isotherms. An analytical solution of the Clausius-Clapeyron equation was proposed to compute the isosteric heat of sorption, the differential entropy, and Gibbs free energy using the Oswin model when the effect of temperature on the hygroscopic equilibrium was considered.

Orange fiber as a novel fat replacer in lemon ice cream

Orange fiber was used as a novel fat replacer in light lemon ice cream. Nine ice cream formulations were compared: standard control ice cream (IC); ice cream with fiber (F1) from the peel, bagasse, and orange seed (ICA and ICB); ice cream with fiber (F2) from the orange peel alone (ICC and ICD); ice cream with fiber (F3) from the peel, bagasse, and orange seed pretreated with hydro-distillation (ICE and ICF); and ice cream with fiber (F4) from the orange peel pretreated with hydro-distillation (ICG and ICH).The orange fiber reduced the ice cream fat content (50 %) and the overrun ratio and increased the fiber content and the hardness, gumminess, and springiness values, but it did not affect the adhesiveness and odor of the samples. The samples with 1.0 % of orange fiber showed low melting rate values than those of the control ice cream. The overall acceptance of the ice cream with 1.0 % of pre-treated orange peel fiber did not differ from that of the control ice cream (80 %). The orange fiber proved a promising food ingredient since it can be used to decrease the fat content and increase bioactive compounds content, such as fiber and carotenoids.

Effects of algal turbidity on foraging and antipredator behaviour of the three-spined stickleback (Gasterosteus aculeatus)

The aim of this thesis was to examine how aquatic organisms, such as fish, behave in an altered environmental condition. Many species of fish use vision as their primary tool to gain information about their surrounding environment. The visual conditions of aquatic habitats are often altered as a result of anthropogenic disturbance, such as eutrophication that initiates algal turbidity. In general, turbidity reduces the visibility and can be hypothesized to have an influence on the behaviour of fish. I used the three-spined stickleback (Gasterosteus aculeatus) as a model species and conducted four studies in the laboratory to test how algal turbidity affects its behaviour. In this thesis, two major behavioural aspects are discussed. The first is antipredator behaviour. In study I, the combined effects of turbidity and shoot density on habitat choice (shelter vs open) behaviour was tested on a group of sticklebacks (20 fish) in the presence and absence of piscivorous perch (Perca fluviatilis). In study II, I examined the behavioural responses of feeding sticklebacks when they were exposed to the sudden appearance of an avian predator (the silhouette of a common tern, Sterna hirundo). The study was done in turbid and clear water using three different groups sizes (1, 3 and 6 fish). The second aspect is foraging behaviour. Study III & IV focused on the effects of algal turbidity on the foraging performance of sticklebacks. In study III, I conducted two separate experiments to examine the effects of turbidity on prey consumption and prey choice of sticklebacks. In this experiment turbidity levels and the proportion of large and small prey (Daphnia spp.) were manipulated. In study IV, I studied whether a group of six sticklebacks can distribute themselves according to food input at two feeding stations in a way that provided each fish with the same amount of food in clear and turbid water. I also observed whether the fish can follow changes in resource distribution between the foraging patches. My results indicate an overall influence of algal turbidity on the antipredator and foraging behaviour of sticklebacks. In the presence of a potential predator, the use of the sheltered habitat was more pronounced at higher turbidity. Besides this, sticklebacks reduced their activity levels with predator presence at higher turbidity and shoot density levels, suggesting a possible antipredator adaptation to avoid a predator. When exposed to a sudden appearance of an avian predator, sticklebacks showed a weaker antipredator response in turbid water, which suggests that turbidity degrades the risk assessment capabilities of sticklebacks. I found an effect of group size but not turbidity in the proportion of sticklebacks that fled to the shelter area, which indicates that sticklebacks are able to communicate among group members at the experimental turbidity levels. I found an overall negative effect of turbidity on food intake. Both turbidity and changes in the proportion of prey sizes played a significant role in a stickleback’s prey selection. At lower turbidity levels (clear <1 and 5 NTU) sticklebacks showed preferences for large prey, whereas in more turbid conditions and when the proportion of large to small prey increased sticklebacks became increasingly random in their prey selection. Finally, my results showed that groups of sticklebacks disperse themselves between feeding stations according to the reward ratios following the predictions of the ideal free distribution theory. However, they took a significantly longer time to reach the equilibrium distribution in turbid water than in clear water. In addition, they showed a slower response to changes in resource distribution in a turbid environment. These findings suggest that turbidity interferes with the information transfer among group foragers. It is important to understand that aquatic animals are often exposed to a degraded environment. The findings of this thesis suggest that algal turbidity negatively affects their behavioural performance. The results also shed light on the underlying behavioural strategies of sticklebacks in turbid conditions that might help them adapt to an altered environmental situation and increase their survival. In conclusion, I hold that although algal turbidity has detrimental effects on the antipredator and foraging behaviour of sticklebacks, their behavioural adjustment might help them adapt to a changing environment.

Influence of agro-ecological production areas on antioxidant activity, reducing sugar content, and selected phytonutrients of orange-fleshed sweet potato cultivars

Sweet potato is an important staple, and it is mainly known for its contribution of β-carotene in human diet. The effects of cultivar and habitat on this pigment and other nutritional characteristics of the crop still require investigation. In this study, three locally bred cultivars of sweet potato, two of which are orange-fleshed, were grown in three different agro-ecological areas to determine soluble sugar content, β-carotene, and total antioxidants of roots. In addition antioxidant activity, total carotenoids, and chlorophyll content were determined in edible leaves. Reducing sugars, β-carotene, total antioxidants capacity, total carotenoids, and chlorophyll content were significantly affected by environmental conditions. The location at lower altitude and closer to the coastline showed high evapotranspiration, thus reducing sugar content, antioxidant activity, and phytonutrients in both storage roots and leaves. Absence of water stress in agro-ecological locations further inland and at higher altitudes was associated with an increase in these compounds. Free radical scavenging activity of DPPH was higher in the storage roots (610.49 µmoles TE/100g) than in the leaves (426.06 µmoles TE/100g); nevertheless, opposite results were found for the ferric ion reducing activity (FRAP). The deep orange-fleshed cultivar A45 contained high β-carotene (15 mg/100g), which is enough to meet RDA for vitamin A. There is evidence of agro-ecological effect on sweet potato nutritional value.

Effects of orange winemaking variables on antioxidant activity and bioactive compounds

AbstractAscorbic acid, carotenoids and polyphenols stand out among the orange juice natural antioxidants. The winemaking process affects their bioavailability and bioactivity. Antioxidant activities (AA) were estimated in different process conditions to asses those properties. The AA and their correlation with ascorbic acid, total phenolics and carotenoids content were calculated. The variables and levels analyzed were: pasteurized and natural must (PJ and NJ), pH 3.5 and 4.0 and fermentation temperatures at 10°C and 20°C. Statistically significant differences (α=0.05) were found among bioactive compounds concentrations. Antioxidant compounds concentration was higher in raw material than in orange wine. Juice pasteurization caused the major losses while subsequent vinification affects them to a lesser extent. Highest antioxidants retention was measured in wines from JN fermented at pH 3.5 and 10 °C (JN-3.5-10) followed by wines from JP and fermented at the same conditions (JP-3.5-10). AA determined by DPPH showed a positive and close correlation with FRAP, while ABTS showed a low correlation with former assays. Juice pasteurization process and fermentation temperature influenced bioactive compound reduction which correlates with the AA variation.

Effects of orange by-product fiber incorporation on the functional and technological properties of pasta

AbstractThe incorporation of fiber into products consumed every day by the general population is important and viable. The aim of the present work was to evaluate the impact of incorporating orange juice industry dietary fiber byproducts in fettuccini of fresh pasta. Three different fiber concentrations were added to fresh pastas (25 g/kg, 50 g/kg and 75 g/kg). The results showed a significant increase in solid loss content when the incorporation of orange fiber was greater than 50 g/kg. This difference did not occur regarding weight increase values and color parameters. The pasta with 75 g/kg orange fiber can be considered a “high fiber” product, with the total dietary fiber content of the pasta increasing by 99% compared to control pasta. The carotenoid and phenolic contents of pasta increased significantly with the incorporation of fiber at 75 g/kg, but only the pasta formulation with 25 g/kg of orange fiber did not differ from control pasta in relation to all of the sensory attributes and presented an acceptance greater than 75%. The addition of orange fiber byproducts to pastas is an interesting alternative because fiber has a high nutritional value and an abundance of antioxidants.