Feeding strategies for <Emphasis Type="Italic">E. coli</Emphasis> fermentations demanding an enriched environment

The addition of a carbon nutrient feed to a fed-batch cultivation is often not enough to obtain satisfactory growth and/or production. In some cases, an additional feed with for example supplementary amino acids or complex media is required. This work presents the development of feeding strategies where more than one feed is required and the knowledge of the growth requirements is low. Simulations and cultivations with E. coli are shown using the proposed feed controllers which are based on a probing control concept. The strategies work well and they can be used to shorten the process development phase considerably.     

Observing growth and division of large numbers of individual bacteria by image analysis

We describe a method that enabled us to observe large numbers of individual bacterial cells during a long period of cell growth and proliferation. We designed a flow chamber in which the cells attached to a transparent solid surface. The flow chamber was mounted on a microscope equipped with a digital camera. The shear force of the flow removed the daughter cells, making it possible to monitor the consecutive divisions of a single cell. In this way, kinetic parameters and their distributions, as well as some physiological characteristics of the bacteria, could be analyzed based on more than 1,000 single-cell observations. The method which we developed enabled us to study the history effect on the distribution of the lag times of single cells.     

Early Stress Causes Sex-Specific,Life-Long Changes in Behaviour,Levels of Gonadal Hormones,and Gene Expression in Chickens

Early stress can have long-lasting phenotypic effects. Previous research shows that male and female chickens differ in many behavioural aspects, and respond differently to chronic stress. The present experiment aimed to broadly characterize long-term sex differences in responses to brief events of stress experienced during the first weeks of life. Chicks from a commercial egg-laying hybrid were exposed to stress by inducing periods of social isolation during their first three weeks of life, followed by a broad behavioural, physiological and genomic characterization throughout life. Early stressed males, but not females, where more anxious in an open field-test, stayed shorter in tonic immobility and tended to have delayed sexual maturity, as shown by a tendency for lower levels of testosterone compared to controls. While early stressed females did not differ from non-stressed in fear and sexual maturation, they were more socially dominant than controls. The differential gene expression profile in hypothalamus was significantly correlated from 28 to 213 days of age in males, but not in females. In conclusion, early stress had a more pronounced long-term effect on male than on female chickens, as evidenced by behavioral, endocrine and genomic responses. This may either be attributed to inherent sex differences due to evolutionary causes, or possibly to different stress related selection pressures on the two sexes during commercial chicken breeding.     

Emergence of Polymorphic Mating Strategies in Robot Colonies

Polymorphism has fascinated evolutionary biologists since the time of Darwin. Biologists have observed discrete alternative mating strategies in many different species. In this study, we demonstrate that polymorphic mating strategies can emerge in a colony of hermaphrodite robots. We used a survival and reproduction task where the robots maintained their energy levels by capturing energy sources and physically exchanged genotypes for the reproduction of offspring. The reproductive success was dependent on the individuals'' energy levels, which created a natural trade-off between the time invested in maintaining a high energy level and the time invested in attracting mating partners. We performed experiments in environments with different density of energy sources and observed a variety in the mating behavior when a robot could see both an energy source and a potential mating partner. The individuals could be classified into two phenotypes: 1) forager, who always chooses to capture energy sources, and 2) tracker, who keeps track of potential mating partners if its energy level is above a threshold. In four out of the seven highest fitness populations in different environments, we found subpopulations with distinct differences in genotype and in behavioral phenotype. We analyzed the fitnesses of the foragers and the trackers by sampling them from each subpopulation and mixing with different ratios in a population. The fitness curves for the two subpopulations crossed at about 25% of foragers in the population, showing the evolutionary stability of the polymorphism. In one of those polymorphic populations, the trackers were further split into two subpopulations: (strong trackers) and (weak trackers). Our analyses show that the population consisting of three phenotypes also constituted several stable polymorphic evolutionarily stable states. To our knowledge, our study is the first to demonstrate the emergence of polymorphic evolutionarily stable strategies within a robot evolution framework.     

Optimizing the Refractive Index Sensitivity of Plasmonically Coupled Gold Nanoparticles

Plasmonics - The possibility to enhance the local refractive index sensitivity using plasmonic coupling between spherical gold nanoparticles (Au-NPs) has been investigated. A strong and distinct...     

The Strong Selective Sweep Candidate Gene ADRA2C Does Not Explain Domestication Related Changes In The Stress Response Of Chickens

Analysis of selective sweeps to pinpoint causative genomic regions involved in chicken domestication has revealed a strong selective sweep on chromosome 4 in layer chickens. The autoregulatory α-adrenergic receptor 2C (ADRA2C) gene is the closest to the selective sweep and was proposed as an important gene in the domestication of layer chickens. The ADRA2C promoter region was also hypermethylated in comparison to the non-selected ancestor of all domesticated chicken breeds, the Red Junglefowl, further supporting its relevance. In mice the receptor is involved in the fight-or-flight response as it modulates epinephrine release from the adrenals. To investigate the involvement of ADRA2C in chicken domestication, we measured gene expression in the adrenals and radiolabeled receptor ligand in three brain regions comparing the domestic White Leghorn strain with the wild ancestor Red Junglefowl. In adrenals ADRA2C was twofold greater expressed than the related receptor gene ADRA2A, indicating that ADRA2C is the predominant modulator of epinephrine release but no strain differences were measured. In hypothalamus and amygdala, regions associated with the stress response, and in striatum, receptor binding pIC50 values ranged between 8.1–8.4, and the level was not influenced by the genotyped allele. Because chicken strains differ in morphology, physiology and behavior, differences attributed to a single gene may be lost in the noise caused by the heterogeneous genetic background. Therefore an F₁₀ advanced intercross strain between White Leghorn and Red Junglefowl was used to investigate effects of ADRA2C alleles on fear related behaviors and fecundity. We did not find compelling genotype effects in open field, tonic immobility, aerial predator, associative learning or fecundity. Therefore we conclude that ADRA2C is probably not involved in the domestication of the stress response in chicken, and the strong selective sweep is probably caused by selection of some unknown genetic element in the vicinity of the gene.     

The effect of thermal stress on protein composition in dogwhelks (Nucella lapillus) under normoxic and hyperoxic conditions

In this laboratory study, dogwhelks (Nucella lapillus) were collected from the intertidal zone and exposed to 16 °C (ambient), 26.5 °C and 30 °C under normal and hyperoxic conditions respectively. It was shown that there was no thermally induced mortality at 26.5 °C, but that the mortality rate was 40–50% in 30 °C. This mortality rate was reduced to 10% if extra oxygen was provided, indicating that oxygen supply was setting the limit for whole organism thermal tolerance. Tissue samples were then analysed for protein features using two-dimensional gel electrophoresis, and both up and down regulation of proteins were visualised by silver staining and crosswise comparisons of gels from control vs. treated animals. The results clearly show that the protein profiles from dogwhelks exposed to increased water temperatures differ from those of the control, but that increased oxygen availability alleviates these differences thus increasing the similarity between heat-shocked and control animal protein pattern. This implies a more stable protein metabolism and might explain the increased survival of heat-shocked individuals when extra oxygen is supplied.     

Regional differences in mRNA responses in blue mussels within the Baltic proper

Mussels (Mytilus sp.) from two regions along the permanent salinity gradient within the Baltic proper were exposed to copper (35 ppb) or petrol (0.3 mL/L) for 10 days and analyzed for mRNA expressions in gill tissue. Expression of mRNAs for the heat shock proteins HSP70 and HSP90 was significantly induced by copper, but not by petrol. For the metallothioneins MT10 and MT20, regional differences in mRNA expressions could be seen. In mussels from the northern Baltic proper, MT20 expression increased 2.8 and 3.4 times, after exposure to copper and petrol, respectively. In contrast, no change could be seen in MT20 expression for mussels from the southern Baltic proper. MT10 showed a peculiar expression not previously described. For some mussels, no expression at all was detected, some showed a weak expression and for some individuals a strong expression could be seen. For the mussels from the southern Baltic proper, the number of individuals with a strong expression of MT10 increased from 1 out of 18 (control), to 7 and 8, after exposure to copper and petrol, respectively. The results clearly show that responses vary between different regions within the Baltic proper, which emphasises the importance to study interactions between contaminants, populations and regions.     

The effect of thermal stress on protein composition in dogwhelks (Nucella lapillus) under normoxic and hyperoxic conditions.

In this laboratory study, dogwhelks (Nucella lapillus) were collected from the intertidal zone and exposed to 16 degrees C (ambient), 26.5 degrees C and 30 degrees C under normal and hyperoxic conditions respectively. It was shown that there was no thermally induced mortality at 26.5 degrees C, but that the mortality rate was 40-50% in 30 degrees C. This mortality rate was reduced to 10% if extra oxygen was provided, indicating that oxygen supply was setting the limit for whole organism thermal tolerance. Tissue samples were then analysed for protein features using two-dimensional gel electrophoresis, and both up and down regulation of proteins were visualised by silver staining and crosswise comparisons of gels from control vs. treated animals. The results clearly show that the protein profiles from dogwhelks exposed to increased water temperatures differ from those of the control, but that increased oxygen availability alleviates these differences thus increasing the similarity between heat-shocked and control animal protein pattern. This implies a more stable protein metabolism and might explain the increased survival of heat-shocked individuals when extra oxygen is supplied.