Alginate embedding and subsequent sporulation of in vitro-produced Conidiobolus thromboides hyphae using a pressurised air-extrusion method

Conidiobolus thromboides is an entomophthoralean fungus with potential as a biological control agent of aphids. However, its application in biological control is limited due to its formulation requirements. The objective of this study was to develop and optimise a novel air-extrusion method to embed C. thromboides hyphae at high density in alginate pellets. An orthogonal experimental design was used to investigate selected combinations of parameters known to affect hyphal density within pellets. The diameter of pellets produced, and the calculated density of hyphae within them, ranged from 0.18 ± 0.09 to 3.17 ± 0.06 mm and from 0.02 to 350.56 mg/mm³ respectively. These data were used to predict the optimal parameter combination to deliver the greatest density of hyphae of C. thromboides per pellet: 1% sodium alginate, a 1:2 ratio of hyphae to sodium alginate, an orifice diameter of 0.232 mm and an air pressure of 0.05 MPa. Pellets made under the optimal conditions predicted produced a mean total of 4.3 ± 0.6 × 10⁵ conidia per pellet at 100% relative humidity which was significantly greater than the mean total number of conidia produced from infected aphid cadavers of comparable size (9.35 ± 0.85 × 10⁴) (p < 0.001). In conclusion, air-extrusion embedding appears to be a promising method for formulating in vitro-produced hyphae of C. thromboides for use in biological control.     

Fitness Is Strongly Influenced by Rare Mutations of Large Effect in a Microbial Mutation Accumulation Experiment

Our understanding of the evolutionary consequences of mutation relies heavily on estimates of the rate and fitness effect of spontaneous mutations generated by mutation accumulation (MA) experiments. We performed a classic MA experiment in which frequent sampling of MA lines was combined with whole genome resequencing to develop a high-resolution picture of the effect of spontaneous mutations in a hypermutator (ΔmutS) strain of the bacterium Pseudomonas aeruginosa. After ∼644 generations of mutation accumulation, MA lines had accumulated an average of 118 mutations, and we found that average fitness across all lines decayed linearly over time. Detailed analyses of the dynamics of fitness change in individual lines revealed that a large fraction of the total decay in fitness (42.3%) was attributable to the fixation of rare, highly deleterious mutations (comprising only 0.5% of fixed mutations). Furthermore, we found that at least 0.64% of mutations were beneficial and probably fixed due to positive selection. The majority of mutations that fixed (82.4%) were base substitutions and we failed to find any signatures of selection on nonsynonymous or intergenic mutations. Short indels made up a much smaller fraction of the mutations that were fixed (17.4%), but we found evidence of strong selection against indels that caused frameshift mutations in coding regions. These results help to quantify the amount of natural selection present in microbial MA experiments and demonstrate that changes in fitness are strongly influenced by rare mutations of large effect.     

Metabolic modeling of single Th17 cells reveals regulators of autoimmunity

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Étude électrophorétique de l'hybride expérimental entre Papio anubis et P. cynocephalus

Hybrids between Papio anubis and P. cynocephalus occur in nature. We have studied laboratory bred hybrids, together with their backcrossed offspring. Electrophoretic experiments on the proteins, serum enzymes and erythrocytes obtained from both hybrids and parents lead to three conclusions. First, both parents and hybrids possess proteins having identical mobilities; this case is the most frequent in the 23 loci studied. Second, individual polymorphism caused by, for example, carbonic anhydrase I or transferrin in one or both parents results in the appearance of a codominant phenotype in the hybrid, when the parental components are distinguishable. Third, when other enzymes such as albumin, alkaline phosphatase, haemoglobin and the serum esterases have particular electrophoretic mobilities in the parents, then the hybrid is again codominant; a particularly interesting case among the esterases is that of pseudocholinesterase, where the paternal component is partially repressed in the hybrid. The hybrid than exhibits the phenomenon of allelic repression of at least one gene, which confirms the taxonomic position of species of the two forms of Papio studied.     

What paths do advantageous alleles take during short‐term evolutionary change?

Combining experimental evolution with whole‐genome resequencing is a promising new strategy for investigating the dynamics of evolutionary change. Published studies that have resequenced laboratory‐selected populations of sexual organisms have typically focused on populations sampled at the end of an evolution experiment. These studies have attempted to associate particular alleles with phenotypic change and attempted to distinguish between different theoretical models of adaptation. However, neither the population used to initiate the experiment nor multiple time points sampled during the evolutionary trajectory are generally available for examination. In this issue of Molecular Ecology, Orozco‐terWengel et al. (2012) take a significant step forward by estimating genome‐wide allele frequencies at the start, 15 generations into and at the end of a 37‐generation Drosophila experimental evolution study. The authors identify regions of the genome that have responded to laboratory selection and describe the temporal dynamics of allele frequency change. They identify two common trajectories for putatively adaptive alleles: alleles either gradually increase in frequency throughout the entire 37 generations or alleles plateau at a new frequency by generation 15. The identification of complex trajectories of alleles under selection contributes to a growing body of literature suggesting that simple models of adaptation, whereby beneficial alleles arise and increase in frequency unimpeded until they become fixed, may not adequately describe short‐term response to selection.     

Invertebrate seedbanks: rehydration of soil from an unregulated river floodplain in the south-eastern U.S.

1. We investigated the responsiveness of aquatic invertebrates to rehydration of floodplain soil in a south‐eastern U.S. river floodplain. Non‐inundated soil divots containing invertebrate seedbanks were collected from three floodplain elevations with different inundation histories (inundated for 2, 38, and 78% of a year), and subjected to rehydration (experimental inundation) in aquaria for 10 weeks. 2. Before rehydration, samples were collected to assess the initial density and composition of invertebrates in non‐inundated soil at each elevation. After rehydration, benthic samples were collected biweekly and emergence of aquatic insects was collected weekly from aquaria. 3. The surviving invertebrates were classified as aquatic, semi‐aquatic and terrestrial. Some aquatic invertebrates survived the non‐inundated period in an active state (e.g. Chironomidae and Ceratopogonidae), while others appeared to be dormant (e.g. Heptageniidae, microcrustaceans and Chironomidae). The response for several invertebrates (e.g. Chironominae, Oligochaeta, Collembola and terrestrial invertebrates) depended on the length of rehydration and inundation history. 4. The capacity of aquatic invertebrates to survive and recover was greatest at the most frequently inundated site. Despite variation in floodplain environments, both active and dormant invertebrates persisting in non‐inundated floodplain soil contributed to the floodplain assemblage during flooding. Thus, maintaining the connection between river and floodplain appears to be important in sustaining the invertebrate seedbank and its capacity to recover during inundation. River channelisation and regulation that severs this connection may result in a loss of diversity and abundance.     

Phosphorylated Thiamine Derivatives and Cortical Activity in the Baboon Papio papio: Effect of Intermittent Light Stimulation

The effect of intermittent light stimulation (ILS) on the distribution of thiamine derivatives in three brain areas (occipital, motor, and premotor) was compared in photosensitive and nonphotosensitive baboons. ILS induces paroxysmal discharges in the motor and premotor areas of photosensitive animals only. In baboons submitted to ILS, thiamine triphosphate (TTP) decreases in both photosensitive and nonphotosensitive animals; thiamine monophosphate (TMP) increases in photosensitive animals, which present ILS-induced paroxysmal discharges, whereas it is unaffected in nonphotosensitive animals. The variations are the most significant in the occipital (visual) cortex. A consumption of TTP may result from electrical activity induced by light stimulation in the occipital area. No correlation between ILS-induced paroxysmal activity and a decrease in TTP contents was found. However, photosensitive animals are affected differently from nonphotosensitive animals, as their content of TMP in the cerebral cortex increases on stimulation. However, as long as the exact role of thiamine compounds in relation to membrane excitability in the nervous system remains unknown, it is impossible to conclude whether the differences observed in the metabolism of thiamine compounds are the cause or the consequence of the photosensitivity in the baboon Papio papio.