Self-assembly of bovine beta-casein below the isoelectric pH

Beta-casein is an intrinsically unstructured amphiphilic protein that self-assembles into micelles at neutral pH. This paper reports that beta-casein self-organizes into micelles also under acidic conditions. The protein association behavior and micelle characteristics at pH 2.6, well below the p I, are presented. The pH was found to strongly affect the micelle shape and dimensions. Cryogenic transmission electron microscopy (cryo-TEM) experiments revealed disk-like micelles of 20-25 nm in length and approximately 3.5 nm in height in acidic conditions. An aggregation number of 6 was determined by sedimentation equilibrium under these conditions. Isothermal titration calorimetry experiments verified the association below the p I and allowed determination of the micellization enthalpy, the critical micellar concentration, and the micellization relative cooperativity (MR). Small-angle X-ray scattering results at concentrations below the critical micellization concentration (CMC) suggest that the monomeric protein is likely in a premolten globule state at low pH. Calculations of the protein charge at acidic and neutral pH reveal a similar high net charge but considerable differences in the charge distribution along the protein backbone. Overall the results show that beta-casein is amphiphilic at low pH, but the distribution of charge along the protein chain creates packing constraints that affect the micelle organization, leading at concentrations above the CMC to the formation of disk micelles.     

Isolation,characterization and effect of plant-growth-promoting rhizobacteria on pine seedlings(Pinus pseudostrobus Lindl.)

In this study,10 bacterial strains were isolated from the rhizosphere of coniferous trees on Mount Tla′loc in Mexico.The strains were characterized by their capacity to produce auxins,solubilize phosphates and stimulate mycelial growth of the ectomycorrhizal fungus Suillus sp.All isolates were identified at the molecular level.Moreover,an experiment was established to evaluate the response of Pinus pseudostrobus seedlings to inoculation with the rhizobacteria strains.The isolated strains belonged to the species Cupriavidus basilensis,Rhodococcus qingshengii,R.erythropolis,Pseudomonas spp.,P.gessardii,Stenotrophomonas rhizophila and Cohnella sp.All of the strains produced auxins;the best producer was R.erythropolis CPT9(76.4 lg mL^-1).P.gessardii CPT6 solubilized phosphate at a significant level(443 lg mL^-1).The strain S.rhizophila CPT8 significantly increased the radial growth of the ectomycorrhizal fungus Suillus sp.by 18.8%.Five strains increased the dry mass of the shoots;R.qingshengii CPT4 and R.erythropolis CPT9 increased growth the most,by more than 20%.Inoculation with plant-growth-promoting rhizobacteria can be a very useful practice in a forest nursery to produce healthy,vigorous plants.     

Mercury in the Aquatic Environment of the Village of Caimito at the Mojana Region,North of Colombia

During May–September 1999, several aquatic environmental samples were collected from the village of Caimito, in the San Jorge River basin area, state of Sucre (Colombia), and analyzed for total mercury (T-Hg). Mean T-Hg concentration in sediments from surrounded marshes was 0.155 ± 0.016 g Hg/g. Low Hg concentrations were found in the phytoplanktivorous fish species Prochilodus magdalenae (0.087 ± 0.01 g Hg/g), while four-fold greater concentrations were found in all the carnivorous species. In average, T-Hg concentrations for fish samples did not exceed the limit consumption level (0.5 g Hg/g). However, risk assessment based on the hazard index suggested that the consumption of 0.1 kg per day of carnivorous fish could increase the risk of mercury poisoning in local population.     

Modelling the effects of land-use change on runoff and sediment yield for a meso-scale catchment in the Southern Pyrenees

The Southern Pre-Pyrenees experienced a substantial land-use change over the second half of the 20th century owing to the reduction of agricultural activities towards the formation of a more natural forest landscape. The land-use change over the last 50 years with subsequent effects on water and sediment export was modelled with the process-based, spatially semi-distributed WASA-SED model for the meso-scale Canalda catchment in Catalonia, Spain. It was forwarded that the model yielded plausible results for runoff and sediment yield dynamics without the need of calibration, although the model failed to reproduce the shape of the hydrograph and the total discharge of several individual rainstorm events, hence the simulation capabilities are not yet considered sufficient for decision-making purposes for land management. As there are only a very limited amount of measured data available on sediment budgets with altered land-use and climate change settings, the WASA-SED model was used to obtain qualitative estimates on the effects of past and future change scenarios to derive a baseline for hypothesis building and future discussion on the evolution of sediment budgets in such a dryland setting. Simulating the effects of the past land-use change, the model scenarios resulted in a decrease of up to 75% of the annual sediment yield, whereas modelled runoff remained almost constant over the last 50 years. The relative importance of environmental change was evaluated by comparing the impact on sediment export of land-use change, that are driven by socio-economic factors, with climate change projections for changes in the rainfall regime. The modelling results suggest that a 20% decrease in annual rainfall results in a decrease in runoff and sediment yield, thus an ecosystem stabilisation in regard to sediment export, which can only be achieved by a substantial land-use change equivalent to a complete afforestation. At the same time, a 20% increase in rainfall causes a large export of water and sediment resources out of the catchment, equivalent to an intensive agricultural use of 100% of the catchment area. For wet years, the effects of agricultural intensification are more pronounced, so that in this case the intensive land-use change has a significantly larger impact on sediment generation than climate change. The WASA-SED model proved capable in quantifying the impacts of actual and potential environmental change, but the reliability of the simulation results is still circumscribed by considerable parameterisation and model uncertainties.     

Genetics of litter size in three maternal lines of rabbits: repeatability versus multiple-trait models

Variance components were estimated in 3 lines of rabbits selected for litter size at weaning (A, Prat, and V) to test one of the assumptions of the models used for selection: that litter size data at different parities are repeated measurements of the same trait. Multiple-trait analyses were performed for the total number of kits born (TB), the number of kits born alive (BA), and the number of kits weaned (NW) per litter. Estimates were obtained by REML in multivariate analyses, including all of the information of the selection criteria, under a repeatability model or a multiple-trait model, considering litter size at the first, second, and subsequent parities as different traits. Models included the fixed effects of the physiological status of the female and the year-season of mating day, buck and doe random permanent environmental effects, and doe additive genetic effects. Results indicated that prolificacy was determined mainly by doe components and that the service sire had a very small effect. Heritabilities for the first and second parities were greater than the estimates obtained under the repeatability model (0.04 to 0.14 for the repeatability model). In the A and V lines, similar values of heritability were found at the first and second parities, but in the Prat line heritability at the second parity was always greater than at the first and greater parities (values of 0.21, 0.17, and 0.15 for TB, BA, and NW, respectively, in second parities of the Prat line). Genetic correlations between the same traits at different parities were approximately 0.8 for all traits in line A, but much lower in the other 2 lines. On average, the values were 0.64 for TB, 0.48 for BA, and 0.39 for NW between the first and second parities, and 0.65 for TB, 0.56 for BA, and 0.45 for NW between the first and third and greater parities. Genetic correlations between the second and greater parities showed the greatest values (approximately 0.8) in lines A and Prat for all traits, but they were lower in line V (0.63 for BA and 0.37 for NW). The heterogeneity of heritabilities and genetic correlations between parities lower than 0.9 suggests that litter size at different parities could be considered as different traits when genetic evaluations are performed. However, when the accuracies of predicted breeding values under a multiple-trait and a repeatability model were calculated, assuming the first to be the true model, the values obtained were nearly the same for all traits in all lines.     

Differences in swimming performance and energetic costs between an endangered native toothcarp (Aphanius iberus) and an invasive mosquitofish (Gambusia holbrooki)

Swimming performance is a key feature that mediates fitness and survival in many fish species. Using a swim tunnel respirometer, we compared prolonged swimming performance and energy use for two competing species: an endangered, endemic toothcarp (Aphanius iberus) and a worldwide invasive mosquitofish (Gambusia holbrooki). Critical (U_crit) and optimal swimming speeds, standard and maximal metabolic rates, absolute aerobic scope, as well as the minimum cost of transport were estimated and compared between species and sexes. Body streamlining and caudal peduncle depth were also measured to explain the differences in swimming performance and efficiency. Both sexes of A. iberus presented similar swimming capacity and metabolic traits, whereas males of G. holbrooki showed higher critical swimming speeds, maximal metabolic rate and absolute aerobic scope than females. We also found marked differences between species in most of the response variables examined. Aphanius iberus showed lower swimming capacity (U_crit mean <10 cm s⁻¹), higher maximal metabolic rate and absolute aerobic scope than the invasive species. By contrast, G holbrooki swam faster and had lower cost of transport at a given fish mass and speed, thereby leading to a higher swimming efficiency. The observed differences in swimming efficiency were closely related to differences in morphological characteristics and therefore to drag pressures and propulsion. Our results add a mechanistic basis to the ecological understanding of these two species and suggest that although both are poor swimmers compared to many other similarly sized species, the native species likely has more restricted water flow tolerance and dispersal capacities.     

Elliptical selection experiment for the estimation of genetic parameters of the growth rate and feed conversion ratio in rabbits

Two elliptical selection experiments were performed in two contemporary sire lines of rabbits (C and R) in order to optimize the experimental design for estimating the genetic parameters of the growth rate (GR) and feed conversion ratio (FCR). Twelve males and 19 females from line C, and 13 males and 23 females from line R, were selected from an ellipse defined by a quadratic index based on these traits. Data from 160 rabbits of each of the parental generations of lines C and R and their offspring (275 and 266 animals, respectively) were used for the analysis. A Bayesian framework was adopted for inference. Marginal posterior distributions of the genetic parameters were obtained by Gibbs sampling. An animal model including batch, parity order, litter size, and common environmental litter effects was assumed. Posterior means (posterior standard deviations) for heritabilities of GR and FCR were estimated to be 0.31 (0.10) and 0.31 (0.10), respectively, in line C and 0.21 (0.08) and 0.25 (0.12) in line R. Posterior means of the proportion of the variance due to common litter environmental effects were 0.14 (0.06) and 0.21 (0.06) for GR and FCR, respectively, in line C and 0.17 (0.06) and 0.22 (0.06) in line R. Posterior means of genetic correlation between both traits were -0.49 (0.25) in line C and -0.47 (0.32) in line R, indicating that selection for GR was expected to result in a similar correlated response in FCR in both lines.     

Genetic parameters of fertility in two lines of rabbits with different reproductive potential

A Bayesian analysis with a threshold model was performed for fertility defined as a binary trait (1 = successful mating, 0 = unsuccessful mating) in two populations of rabbits of different reproductive potential and different genetic origin: Line P selected for litter size and Line C selected for growth rate. There were 20,793 records of natural mating (86.2% successful) in Line C between 1983 and 2003, and 17,548 records (80.5% successful) in Line P, between 1992 and 2003. Data related to 5,388 and 3,848 females and 1,021 and 685 males in Lines C and P, respectively. The pedigree included 6,409 and 4,533 individuals in Lines C and P, respectively. The binary response was modeled under a probit approach. The model for the latent variable included male and female additive genetic effects, male and female permanent environmental effects, and the year-season and physiological status of the female (nulliparous, multiparous lactating, or multiparous nonlactating) as systematic effects. Means (standard deviation in parentheses) of the estimated marginal posterior distribution (EMPD) of male heritability were 0.013 (0.006) and 0.010 (0.008) in Lines C and P, respectively, and those of EMPD of female heritability were 0.056 (0.013) and 0.062 (0.018) in Lines C and P, respectively. Means of the EMPD of the proportion of the phenotypic variance due to environmental male and female effects were, respectively, 0.031 (0.007) and 0.128 (0.018) in Line C and 0.053 (0.010) and 0.231 (0.024) in Line P. Means (standard deviations in parentheses) of the EMPD of genetic correlation between male and female fertility were 0.733 (0.197) in Line C and 0.434 (0.381) in Line P. The posterior distribution of genetic correlations presents a huge dispersion, and the estimates should be taken with caution because of the almost negligible estimate of the male genetic component. Results indicate that little genetic variation exists for female fertility, and practically none for male fertility. It would, therefore, be possible to improve reproductive performance by including female fertility in a breeding program, but response to selection would be very small.     

Xenotransplantation of goat ovary as an alternative to analyse follicles after vitrification

The aim of this study was to evaluate the caprine preantral follicles enclosed on vitrified/warmed ovarian cortex grafted to nude BALB/mice during 1 month. The ovarian cortex from goats was fragmented (3 × 3 × 0.5 mm) and divided into four groups: fresh control, vitrified control, fresh transplant and vitrified transplant. Follicular morphology, development and density, fibrosis as well as apoptosis, and tissue revascularization were evaluated. It was also observed a significant decrease in morphologically normal preantral (primordial, transition, primary and secondary) follicles in both vitrified control and vitrified transplant treatments when compared with both fresh control and fresh transplant. However, fresh control and fresh transplant exhibited a similar percentage of developing follicles. Additionally, Vitrified control showed a significant increase in developing follicles in comparison with both fresh control and fresh transplant. Follicular density significantly decreased in all treatments in comparison with fresh control. We observed high fibrosis in both fresh transplant and vitrified transplant. The mRNA expression of caspase 3 was lower in both fresh transplant and vitrified transplant in comparison with vitrified control. In conclusion, xenotransplantation is an excellent strategy to maintain normal preantral follicle morphology after vitrification/warming of goat ovarian tissue. Yet, in order to ensure the survival and development of these follicles, it is essential to improve the revascularization of the graft.