The Influence of Parasite Infection on Mating Success in Spadefoot Toads, Scaphiopus couchii

The desert toads, Scaphiopus couchii, have an annual activityseason of less than 8 weeks and experience only one significantparasite infection: the monogenean Pseudodiplorchis americanusis transmitted during host spawning and provides a natural systemfor testing the influence of parasite burden on host matingsuccess. The 10 month hibernation involves total starvationduring which the blood-feeding parasites reduce fat reservesand haematocrit. The toads emerge and spawn on the first nightafter rainfall, before they replenish depleted reserves. Malechorusing is energetically very demanding and mate selection,limited to a 7 hr nocturnal assembly, is determined principallyby female choice. Parasite transmission triggered by host sexualactivity results in 100% prevalence and high intensities ofinfection amongst males. Around 50% of toads lose their burdensentirely but the rest carry chronic infections throughout hosthibernation. Field data show a consistent reduction in intensityeach year strongly suggesting resistance. Parasite infectionis pathogenic and creates extra stress during hibernation; therefore,to the extent that elimination of infection is heritable, toadsentering spawning assemblies with heavy burdens should makepoor mates. However, extensive field studies show no correlationbetween mate success and parasite burden. Although infectioncan prejudice survival, it is only one of several inter-relatedfactors (including feeding success, tolerance of hibernation).The condition of successful and unsuccessful males in spawningassemblies indicates that all exceed a threshold at which parasite-inducedpathology is significant. Males which are debilitated by infection—orother factors—are selected against before mate choicebegins     

Preservation of Human Tracks in Arid Environments

Observations were made on the preservation of modern human tracks in a salina near Cuatro Cienegas, Coahuila, Mexico. Track making potential is relatively high in such arid, ephemeral lake basin settings where salinas or alkali flats typically develop. However, because of the instability of evaporite minerals, long-term preservation potential of such tracks is compromised. The substrate in the study area is composed of a mixture of sand and a high percentage of sulfate crystals (gypsum, bloedite, epsomite). At the sediment-air interface, there is a thin white, crystalline sulfate crust that overlies the original track-bearing surface and prevents water loss. The size and surface growth of sulfate crystals in the crust is affected by the differential pressure caused by the track maker. Such tracks represent a good example of a biogenic process (track formation) affecting a physical sedimentological process.     

Inter-Annual Precipitation Variability Decreases Switchgrass Productivity from Arid to Mesic Environments

Cellulosic biofuels are an important source of renewable biomass within the alternative energy portfolio. Switchgrass (Panicum virgatum L.), a perennial C₄ grass native to North America, is widely studied as a biofuel feedstock for its consistently high yields and minimal input requirements. The influences of precipitation amount and temporal variability on the fertilizer response of switchgrass productivity are not fully understood. Moreover, global climate models predict changes in rainfall patterns towards lower and increasingly variable soil water availability in several productive areas worldwide, which may impact net primary production of biofuel crops. We conducted a meta-analysis of aboveground net primary production of switchgrass from 48 publications encompassing 82 different locations, 11 soil types, 52 switchgrass cultivars, fertilizer inputs between 0 to 896 kg N ha^?1 year^?1, and 1 to 6 years of annual productivity measures repeated on the same stand. Productivity of the lowland ecotype doubled with N rates >?131 kg N ha^?1 year^?1, but upland ecotype productivity increased only by 50%. Results showed an optimum N rate of 30 to 60 kg N ha^?1 year^?1 for both ecotypes, after which biomass gain per unit of N added decreased. Growing season precipitation (GSPPT) and inter-annual precipitation variability (inter-PPTvar) affected both ecotypes similarly. Long-term mean annual precipitation (MAP) differentially affected lowland and upland productivity, depending on the N level. Productivity responses to MAP and GSPPT were similar for both upland and lowland ecotypes at none or low N rates. When N increased beyond 60 kg N ha^?1 year^?1, lowland cultivars had a greater growth response to MAP than uplands. Productivity increased with increasing GSPPT and MAP and had a positive linear response to MAP ranging from 600 to 1200 mm year^?1. One third of the variability in switchgrass production was accounted for by inter-PPTvar. After accounting for MAP, sites with higher inter-PPTvar had lower switchgrass productivity than sites with lower inter-PPTvar. Increased inter-annual variation in precipitation reduced production of both ecotypes. Predicted changes in the amount and timing of precipitation thus likely will exert greater influence on production of upland than lowland ecotypes of switchgrass.     

Improved Climate Risk Simulations for Rice in Arid Environments

We integrated recent research on cardinal temperatures for phenology and early leaf growth, spikelet formation, early morning flowering, transpirational cooling, and heat- and cold-induced sterility into an existing to crop growth model ORYZA2000. We compared for an arid environment observed potential yields with yields simulated with default ORYZA2000, with modified subversions of ORYZA2000 and with ORYZA_S, a model developed for the region of interest in the 1990s. Rice variety ‘IR64’ was sown monthly 15-times in a row in two locations in Senegal. The Senegal River Valley is located in the Sahel, near the Sahara desert with extreme temperatures during day and night. The existing subroutines underestimated cold stress and overestimated heat stress. Forcing the model to use observed spikelet number and phenology and replacing the existing heat and cold subroutines improved accuracy of yield simulation from EF = −0.32 to EF =0.70 (EF is modelling efficiency). The main causes of improved accuracy were that the new model subversions take into account transpirational cooling (which is high in arid environments) and early morning flowering for heat sterility, and minimum rather than average temperature for cold sterility. Simulations were less accurate when also spikelet number and phenology were simulated. Model efficiency was 0.14 with new heat and cold routines and improved to 0.48 when using new cardinal temperatures for phenology and early leaf growth. The new adapted subversion of ORYZA2000 offers a powerful analytic tool for climate change impact assessment and cropping calendar optimisation in arid regions.     

Rapid Decline of Ceftazidime Resistance in Antibiotic-Free and Sublethal Environments Is Contingent on Genetic Background

Trade-offs of antibiotic resistance evolution, such as fitness cost and collateral sensitivity (CS), could be exploited to drive evolution toward antibiotic susceptibility. Decline of resistance may occur when resistance to other drug leads to CS to the first one and when compensatory mutations, or genetic reversion of the original ones, reduce fitness cost. Here we describe the impact of antibiotic-free and sublethal environments on declining ceftazidime resistance in different Pseudomonas aeruginosa resistant mutants. We determined that decline of ceftazidime resistance occurs within 450 generations, which is caused by newly acquired mutations and not by reversion of the original ones, and that the original CS of these mutants is preserved. In addition, we observed that the frequency and degree of this decline is contingent on genetic background. Our results are relevant to implement evolution-based therapeutic approaches, as well as to redefine global policies of antibiotic use, such as drug cycling.     

Tumor Stiffness Is Unrelated to Myosin Light Chain Phosphorylation in Cancer Cells

Many tumors are stiffer than their surrounding tissue. This increase in stiffness has been attributed, in part, to a Rho-dependent elevation of myosin II light chain phosphorylation. To characterize this mechanism further, we studied myosin light chain kinase (MLCK), the main enzyme that phosphorylates myosin II light chains. We anticipated that increases in MLCK expression and activity would contribute to the increased stiffness of cancer cells. However, we find that MLCK mRNA and protein levels are substantially less in cancer cells and tissues than in normal cells. Consistent with this observation, cancer cells contract 3D collagen matrices much more slowly than normal cells. Interestingly, inhibiting MLCK or Rho kinase did not affect the 3D gel contractions while blebbistatin partially and cytochalasin D maximally inhibited contractions. Live cell imaging of cells in collagen gels showed that cytochalasin D inhibited filopodia-like projections that formed between cells while a MLCK inhibitor had no effect on these projections. These data suggest that myosin II phosphorylation is dispensable in regulating the mechanical properties of tumors.     

Nonfermentative Thermoalkaliphilic Growth Is Restricted to Alkaline Environments

Caldalkalibacillus thermarum strain TA2.A1 grew in pH-controlled batch culture containing a fermentable growth substrate (i.e., sucrose) from pH 7.5 to 10.0 with no significant change in the specific growth rate, suggesting that this bacterium was a facultative alkaliphile. However, when strain TA2.A1 was grown on a nonfermentable carbon source, such as succinate or malate, no growth was observed until the external pH was >9.0, suggesting that this bacterium was an obligate alkaliphile. Succinate transport and sucrose transport by strain TA2.A1 showed pH profiles similar to that of growth on these carbon sources, and the molar growth yield on sucrose was higher at pH 9.5 than at pH 7.5, despite the increased energy demands on the cell for intracellular pH regulation. Succinate transport, succinate-dependent oxygen consumption, and succinate dehydrogenase and F₁F_o-ATPase specific activities were all significantly lower in cultures of strain TA2.A1 grown at pH 7.5 than in those cultured at pH 9.5. No significant ATP synthesis via the F₁F_o-ATP synthase was detected until the external pH was >8.5. On the basis of these results, we propose that nonfermentative thermoalkaliphilic growth is specialized to function at high pH values, but not at pH values near neutral pH.Alkaliphilic microorganisms have been isolated from a diverse range of environments and have traditionally been classified into two distinct groups based on their pH profile for growth (8). Bacteria that grow across a broad pH range from 7.0 to 11.0 have been classified as facultative alkaliphiles (e.g., Bacillus pseudofirmus OF4) (28), and those that are able to grow only above pH 9.0 have been classified as obligate alkaliphiles (e.g., Bacillus alcalophilus) (4). The reasons why obligate alkaliphiles fail to grow below pH 9.0 remain speculative.While the classification of alkaliphilic bacteria based on pH profiles for growth has gained universal acceptance, it does not consider the nature of the carbon source that is used to grow the cells, and for aerobic alkaliphiles, this may have important consequences. For example, growth on succinate in aerobic bacteria is strictly coupled to oxidative phosphorylation and ATP is produced in the cell via the membrane-bound F₁F_o-ATP synthase. Growth on fermentable carbon sources, such as glucose, allows the cells to bypass this machinery, as ATP can be produced via substrate-level phosphorylation and incomplete oxidation of glucose to acetate can occur.A thermoalkaliphilic bacterium, Bacillus sp. strain TA2.A1, capable of optimal aerobic growth at a temperature of 65°C at pH 9.5 was isolated from an alkaline thermal bore at Mt. Te Aroha, New Zealand (19). The 16S rRNA gene sequence of strain TA2.A1, compared with those available in the EMBL database, shows 99.5% similarity to Caldalkalibacillus thermarum strain HA6^T, an aerobic, heterotrophic, thermophilic bacterium isolated from an alkaline hot spring in China (30). On the basis of the similarity of its phenotypic and genotypic characteristics to those of strain HA6^T, we assign strain TA2.A1 to the genus and species Caldalkalibacillus thermarum. C. thermarum strain TA2.A1 grows on sucrose, common C₄-dicarboxylates, glutamate, pyruvate, and trehalose; however, glucose and fructose fail to support growth (19). We originally described strain TA2.A1 as a facultative alkaliphile based on its pH profile for growth on glutamate or sucrose (18-20); however, both are substrates whose metabolism is not strictly coupled to oxidative phosphorylation.In this communication, we determine the pH profile for growth of C. thermarum strain TA2.A1 on nonfermentable (i.e., succinate and malate) and fermentable carbon sources (i.e., sucrose) using pH-controlled batch culture and demonstrate that strain TA2.A1 was unable to grow below pH 9.0 in pH-controlled batch culture on succinate but grew from pH 7.5 to 10 on sucrose. The physiological and biochemical bases for this phenomenon were investigated.     

Molybdenum Availability Is Key to Nitrate Removal in Contaminated Groundwater Environments

The concentrations of molybdenum (Mo) and 25 other metals were measured in groundwater samples from 80 wells on the Oak Ridge Reservation (ORR) (Oak Ridge, TN), many of which are contaminated with nitrate, as well as uranium and various other metals. The concentrations of nitrate and uranium were in the ranges of 0.1 μM to 230 mM and <0.2 nM to 580 μM, respectively. Almost all metals examined had significantly greater median concentrations in a subset of wells that were highly contaminated with uranium (≥126 nM). They included cadmium, manganese, and cobalt, which were 1,300- to 2,700-fold higher. A notable exception, however, was Mo, which had a lower median concentration in the uranium-contaminated wells. This is significant, because Mo is essential in the dissimilatory nitrate reduction branch of the global nitrogen cycle. It is required at the catalytic site of nitrate reductase, the enzyme that reduces nitrate to nitrite. Moreover, more than 85% of the groundwater samples contained less than 10 nM Mo, whereas concentrations of 10 to 100 nM Mo were required for efficient growth by nitrate reduction for two Pseudomonas strains isolated from ORR wells and by a model denitrifier, Pseudomonas stutzeri RCH2. Higher concentrations of Mo tended to inhibit the growth of these strains due to the accumulation of toxic concentrations of nitrite, and this effect was exacerbated at high nitrate concentrations. The relevance of these results to a Mo-based nitrate removal strategy and the potential community-driving role that Mo plays in contaminated environments are discussed.     

Rapid Intrahost Evolution of Human Cytomegalovirus Is Shaped by Demography and Positive Selection

Populations of human cytomegalovirus (HCMV), a large DNA virus, are highly polymorphic in patient samples, which may allow for rapid evolution within human hosts. To understand HCMV evolution, longitudinally sampled genomic populations from the urine and plasma of 5 infants with symptomatic congenital HCMV infection were analyzed. Temporal and compartmental variability of viral populations were quantified using high throughput sequencing and population genetics approaches. HCMV populations were generally stable over time, with ∼88% of SNPs displaying similar frequencies. However, samples collected from plasma and urine of the same patient at the same time were highly differentiated with approximately 1700 consensus sequence SNPs (1.2% of the genome) identified between compartments. This inter-compartment differentiation was comparable to the differentiation observed in unrelated hosts. Models of demography (i.e., changes in population size and structure) and positive selection were evaluated to explain the observed patterns of variation. Evidence for strong bottlenecks (>90% reduction in viral population size) was consistent among all patients. From the timing of the bottlenecks, we conclude that fetal infection occurred between 13–18 weeks gestational age in patients analyzed, while colonization of the urine compartment followed roughly 2 months later. The timing of these bottlenecks is consistent with the clinical histories of congenital HCMV infections. We next inferred that positive selection plays a small but measurable role in viral evolution within a single compartment. However, positive selection appears to be a strong and pervasive driver of evolution associated with compartmentalization, affecting ≥34 of the 167 open reading frames (∼20%) of the genome. This work offers the most detailed map of HCMV in vivo evolution to date and provides evidence that viral populations can be stable or rapidly differentiate, depending on host environment. The application of population genetic methods to these data provides clinically useful information, such as the timing of infection and compartment colonization.     

Evolution of Integrated Causal Structures in Animats Exposed to Environments of Increasing Complexity

Natural selection favors the evolution of brains that can capture fitness-relevant features of the environment''s causal structure. We investigated the evolution of small, adaptive logic-gate networks (“animats”) in task environments where falling blocks of different sizes have to be caught or avoided in a ‘Tetris-like’ game. Solving these tasks requires the integration of sensor inputs and memory. Evolved networks were evaluated using measures of information integration, including the number of evolved concepts and the total amount of integrated conceptual information. The results show that, over the course of the animats'' adaptation, i) the number of concepts grows; ii) integrated conceptual information increases; iii) this increase depends on the complexity of the environment, especially on the requirement for sequential memory. These results suggest that the need to capture the causal structure of a rich environment, given limited sensors and internal mechanisms, is an important driving force for organisms to develop highly integrated networks (“brains”) with many concepts, leading to an increase in their internal complexity.     

Characterization of Insecticidal Genes of Bacillus thuringiensis Strains Isolated from Arid Environments

This study aimed at characterizing the insecticidal genes of eight Bacillus thuringiensis isolates that were recovered from the local environment of western Saudi Arabia. The screening for the presence of lepidopteran-specific cry1A family and vip3A genes, dipteran-specific cry4 family and coleopteran-specific cry3A, vip1A and vip2A genes, was carried out by PCR. All eight isolates produced PCR products that confirmed the presence of cry1Aa, cry1Ab, cry1Ac, cry4A, cry4B genes, but not cry3A, vip1A and vip2A genes. However, three isolates only were found to carry vip3A genes as revealed by PCR. The observation of cry1 and cry4 genes suggests that these eight isolates may have dual activity against Lepidoptera and Diptera species, while three isolates possessed vip3 genes in addition to cry1 and cry4 which suggests that these three isolates have toxic crystals and vegetative proteins. The results of this study are interesting in the sense that they may help developing new strategies for controlling insects of economic and medical importance in Saudi Arabia, using B. thuringiensis strains that naturally exist in the local environment instead of the current control strategies that are based solely on chemical insecticides.     

Dragonfly Assemblages in Arid Tropical Environments: A Case Study from Western Namibia

Dragonflies have been proposed as indicators for the ecosystem health of freshwater wetlands. For their useful functioning as indicators it is, however, necessary to identify species compositions in specific habitats and species-habitat associations, particularly in the tropics, where such knowledge is still weak. We examined the dragonfly species composition of 133 localities in the arid environment of western Namibia. An analysis of nestedness indicated that distinct, and predictable patterns of species associations can be expected. Discriminant analyses revealed that most of the nine habitat types separated by structural and hydrological parameters are well discriminated by their dragonfly assemblages. Spring brooks in particular host a specific assemblage, which is threatened due to the habitat restriction of several species, as well as by recent habitat loss and degradation. Using a hierarchical method of several criteria we demonstrated the selection of a set of potential indicator species from the species set, most of these being useful indicators for spring brook assemblages. The conservation status of certain habitats and species is discussed. We propose that dragonflies will have a high indicator potential for threatened freshwater wetlands in such areas and may also serve as an indication of the sustainable use of water resources including evaluating measures to rehabilitate environments.     

Patterns of Reproductive Isolation in Toads

Understanding the general features of speciation is an important goal in evolutionary biology, and despite significant progress, several unresolved questions remain. We analyzed an extensive comparative dataset consisting of more than 1900 crosses between 92 species of toads to infer patterns of reproductive isolation. This unique dataset provides an opportunity to examine the strength of reproductive isolation, the development and sex ratios of hybrid offspring, patterns of fertility and infertility, and polyploidization in hybrids all in the context of genetic divergence between parental species. We found that the strength of intrinsic postzygotic isolation increases with genetic divergence, but relatively high levels of divergence are necessary before reproductive isolation is complete in toads. Fertilization rates were not correlated to genetic divergence, but hatching success, the number of larvae produced, and the percentage of tadpoles reaching metamorphosis were all inversely related with genetic divergence. Hybrids between species with lower levels of divergence developed to metamorphosis, while hybrids with higher levels of divergence stopped developing in gastrula and larval stages. Sex ratios of hybrid offspring were biased towards males in 70% of crosses and biased towards females in 30% of crosses. Hybrid females from crosses between closely related species were completely fertile, while approximately half (53%) of hybrid males were sterile, with sterility predicted by genetic divergence. The degree of abnormal ploidy in hybrids was positively related to genetic divergence between parental species, but surprisingly, polyploidization had no effect on patterns of asymmetrical inviability. We discuss explanations for these patterns, including the role of Haldane''s rule in toads and anurans in general, and suggest mechanisms generating patterns of reproductive isolation in anurans.     

Memory in Elementary School Children Is Improved by an Unrelated Novel Experience

Education is the most traditional means with formative effect on the human mind, learning and memory being its fundamental support. For this reason, it is essential to find different strategies to improve the studentś performance. Based on previous work, we hypothesized that a novel experience could exert an enhancing effect on learning and memory within the school environment. Here we show that novel experience improved the memory of literary or graphical activities when it is close to these learning sessions. We found memory improvements in groups of students who had experienced a novel science lesson 1 hour before or after the reading of a story, but not when these events were 4 hours apart. Such promoting effect on long-term memory (LTM) was also reproduced with another type of novelty (a music lesson) and also after another type of learning task (a visual memory). Interestingly, when the lesson was familiar, it failed to enhance the memory of the other task. Our results show that educationally relevant novel events experienced during normal school hours can improve LTM for tasks/activities learned during regular school lessons. This effect is restricted to a critical time window around learning and is particularly dependent on the novel nature of the associated experience. These findings provide a tool that could be easily transferred to the classroom by the incorporation of educationally novel events in the school schedule as an extrinsic adjuvant of other information acquired some time before or after it. This approach could be a helpful tool for the consolidation of certain types of topics that generally demand a great effort from the children.     

Rhodopsin Molecular Evolution in Mammals Inhabiting Low Light Environments

The ecological radiation of mammals to inhabit a variety of light environments is largely attributed to adaptive changes in their visual systems. Visual capabilities are conferred by anatomical features of the eyes as well as the combination and properties of their constituent light sensitive pigments. To test whether evolutionary switches to different niches characterized by dim-light conditions coincided with molecular adaptation of the rod pigment rhodopsin, we sequenced the rhodopsin gene in twenty-two mammals including several bats and subterranean mole-rats. We compared these to thirty-seven published mammal rhodopsin sequences, from species with divergent visual ecologies, including nocturnal, diurnal and aquatic groups. All taxa possessed an intact functional rhodopsin; however, phylogenetic tree reconstruction recovered a gene tree in which rodents were not monophyletic, and also in which echolocating bats formed a monophyletic group. These conflicts with the species tree appear to stem from accelerated evolution in these groups, both of which inhabit low light environments. Selection tests confirmed divergent selection pressures in the clades of subterranean rodents and bats, as well as in marine mammals that live in turbid conditions. We also found evidence of divergent selection pressures among groups of bats with different sensory modalities based on vision and echolocation. Sliding window analyses suggest most changes occur in transmembrane domains, particularly obvious within the pinnipeds; however, we found no obvious pattern between photopic niche and predicted spectral sensitivity based on known critical amino acids. This study indicates that the independent evolution of rhodopsin vision in ecologically specialised groups of mammals has involved molecular evolution at the sequence level, though such changes might not mediate spectral sensitivity directly.     

Retrotransposon mdg3 Is Transferred between Somatic Cells of Unrelated Species in Mixed Culture

Since retrovirus-like particles of gypsy (mdg4) are capable of interspecific transfer, other Drosophila melanogaster gypsy-related retrotransposons were tested for this property. As a donor and a recipient, D. melanogaster and D. virilis cultured cells were used. Recipient cell DNA was analyzed with probes directed to mdg1, mdg3, 17.6, 297, 412, or B104/roo. Transfer was demonstrated for mdg3, which lacks env. The possible mechanism of transfer is discussed.     

Rapid Evolution of Novel Traits in Microorganisms

The use of natural microorganisms in biotransformations is frequently constrained by their limited tolerance to the high concentrations of metabolites and solvents required for effective industrial production. In many cases, more robust strains have to be generated by random mutagenesis and selection. This process of directed evolution can be accelerated in mutator strains, which carry defects in one or more of their DNA repair genes. However, in order to use mutator strains, it is essential to restore the normal low mutation rate of the selected organisms immediately after selection to prevent the accumulation of undesirable spontaneous mutations. To enable this process, we constructed temperature-sensitive plasmids that temporarily increase the mutation frequency of their hosts by 20- to 4,000-fold. Under appropriate selection pressure, microorganisms transformed with mutator plasmids can be quickly evolved to exhibit new, complex traits. By using this approach, we were able to increase the tolerance of three bacterial strains to dimethylformamide by 10 to 20 g/liter during only two subsequent transfers. Subsequently, the evolved strains were returned to their normal low mutation rate by curing the cells of the mutator plasmids. Our results demonstrate a new and efficient method for rapid strain improvement based on in vivo mutagenesis.