Natural variation of gliding motility in a centimetre-scale population of Myxococcus xanthus

A major challenge in microbial evolutionary ecology is to understand how fitness-related traits vary in natural populations of microorganisms at defined spatial scales and subsequently to identify the forces that maintain such variation. The Gram-negative soil bacterium Myxococcus xanthus is a model system for the study of gliding motility, which is driven by two complementary motility systems in this species and is central to its social lifestyle. We tested whether the ecological context of a centimetre-scale M. xanthus population allows the coexistence of diverse motility-related phenotypes. Swarming rates among 26 clones isolated at the centimetre scale were found to vary greatly in multiple laboratory environments. This variation appears to be motility-specific, as it is not explained by a correlated variation in intrinsic growth rate. In contrast to the common reference strain DK1622, most isolates swarmed faster on hard agar than on soft agar, highlighting the difficulty of inferring species characteristics from laboratory reference strains. These isolates also varied greatly in swarm morphology and in the effect of nutrient limitation on swarming rate. Our results show that diverse swarming phenotypes can coexist in a small-scale bacterial population.     

Toxoplasma gondii Hsp20 is a stripe-arranged chaperone-like protein associated with the outer leaflet of the inner membrane complex

Background information. Toxoplasma gondii is among the most successful parasites, with nearly half of the human population chronically infected. T. gondii has five sHsps [small Hsps (heat‐shock proteins)] located in different subcellular compartments. Among them, Hsp20 showed to be localized at the periphery of the parasite body. sHsps are widespread, constituting the most poorly conserved family of molecular chaperones. The presence of sHsps in membrane structures is unusual. Results. The localization of Hsp20 was further analysed using high‐resolution fluorescent light microscopy as well as electron microscopy, which revealed that Hsp20 is associated with the outer surface of the IMC (inner membrane complex), in a set of discontinuous stripes following the same spiralling trajectories as the subpellicular microtubules. The detergent extraction profile of Hsp20 was similar to that of GAP45 [45 kDa GAP (gliding‐associated protein)], a glideosome protein associated with the IMC, but was different from that of IMC1 protein. Although we were unable to detect interacting protein partners of Hsp20 either in normal or stressed tachyzoites, an interaction of Hsp20 with phosphatidylinositol 4‐phosphate and phosphatidylinositol 4,5‐bisphosphate phospholipids could be observed. Conclusions. Hsp20 was shown to be associated with a specialized membranous structure of the parasite, the IMC. This discontinuous striped‐arrangement is unique in T. gondii, indicating that the topology of the outer leaflet of the IMC is not homogeneous.     

Destructive and non-destructive measurements of residual crop residue and phosphorus effects on growth and composition of herbaceous fallow species in the Sahel

Little is known about the residual effects of crop residue (CR) and phosphorus (P) application on the fallow vegetation following repeated cultivation of pearl millet [Pennisetum glaucum (L.) R. Br.] in the Sahel. The objective of this study, therefore, was (i) to measure residual effects of CR, mulched at annual rates of 0, 500, 1000 and 2000 kg CR ha^–1, broadcast P at 0 and 13 kg P ha^–1 and P placement at 0, 1, 3, 5 and 7 kg P ha^–1 on the herbaceous dry matter (HDM) 2 years after the end of the experiment and (ii) to test a remote sensing method for the quantitative estimation of HDM. Compared with unmulched plots, a doubling of HDM was measured in plots that had received at least 500 kg CR ha^–1. Previous broadcast P application led to HDM increases of 14% compared with unfertilised control plots, whereas no residual effects of P placement were detected. Crop residue and P treatments caused significant shifts in flora composition. Digital analysis of colour photographs taken of the fallow vegetation and the bare soil revealed that the number of normalised green band pixels averaged per plot was highly correlated with HDM (r = 0.86) and that red band pixels were related to differences in soil surface crusting. Given the traditional use of fallow vegetation as fodder, the results strongly suggest that for the integrated farming systems of the West African Sahel, residual effects of soil amendments on the fallow vegetation should be included in any comprehensive analysis of treatment effects on the agro-pastoral system.     

COMPARISON OF METHODS USED TO ESTIMATE NUMBERS OF WALRUSES ON SEA ICE

The US and former USSR conducted joint surveys of Pacific walruses on sea ice and at land haul-outs in 1975, 1980, 1985, and 1990. One of the difficulties in interpreting results of these surveys has been that, except for the 1990 survey, the Americans and Soviets used different methods for estimating population size from their respective portions of the sea ice data. We used data exchanged between Sovier and American scientists to compare and evaluate the two estimation procedures and to derive a set of alternative estimates from the 1975, 1980, and 1985 surveys based on a single consistent procedure. Estimation method had only a small effect on total population estimates because most walruses were found at land haul-outs. However, the Sovievr method is subject to bias that depends on the distribution of the population on the sea ice and this has important implications for interpreting the ice portions of previously reported surveys for walruses and other pinniped species. We recommend that the American method be used in future surveys. Future research on survey methods for walruses should focus on other potential sources of bias and variation.     

AERIAL VIDEOGRAMMETRY FROM A TETHERED AIRSHIP TO ASSESS MANATEE LIFE-STAGE STRUCTURE

Aerial videogrammetry from an airship tethered to a boat was used to assess the life-stage structure of manatees in the Blue Waters area of the Homosassa River on the west coast of central Florida. Individual frames of videos were loaded onto a computer and lengths of manatees measured using graphics software. All images of manatees were measured independently by three observers. Measurements were transformed to total manatee lengths by calculating the ratio between the measurements of a reference object of known length and the measurement of the manatee. Length estimates were highly variable but were unbiased with respect to observer. Results of an analysis of variance suggested that the hypothesis that lengths differed among days and among observers should not be rejected. Life-stage structures were represented in three schemes. One scheme-which included four life stages: dependent calves, juveniles, animals in tramsition between juvenile and adult, and adults-was included to provide the most information without sacrificing confidence in the life-stage distribution. Overall, the video system was effective at capturing images of manatees for life-stage characterization and, with improvements in image resolution, could become a valuable tool for photo-identification in sight-resight experiments.     

Anti-Predator Behavioral Responses of Mosquito Pupae to Aerial Predation Risk

Aquatic insects have two potential sources of predation risk: aquatic predators and aerial predators. Our goal was to assess anti-predator responses of Culex pipiens to aerial predation. By simulating predator attacks, we assessed (a) the distance fled in relation to depth and group size, (b) the distribution of individuals at different depths, and (c) the duration of surfacing events to obtain air in scenarios with varying predation risk. Pupae located closer to the surface fled deeper into the water, and the number of conspecifics decreased the distance fled. When the risk of predation increased, more individuals were found deeper in the water column, and the interval between two consecutive surfacing events increased. Culex pipiens shows a trade-off between avoiding aerial predation and maintaining oxygen acquisition, which may be regulated by the need to conserve energy reserves.     

Multiple selection pressures influence Trinidadian guppy (Poecilia reticulata) antipredator behavior

Animals alter their behavior to avoid a variety of differenttypes of predators. Trinidadian guppies (Poecilia reticulata)have been an important system for examining the evolution ofantipredator behavior because geographically isolated populationsexperience different amounts of aquatic predation. Althoughthe influence of aquatic predators has been well documented,selective pressures from other types of predators are less wellunderstood. We examined the response of wild-caught individualsfrom upstream and downstream populations to an aquatic predatorand a simulated aerial predator. As previously documented, fishfrom the downstream population responded more strongly to theaquatic predator than did fish from the upstream population,inspecting for longer periods of time. Guppies also exhibiteda strong behavioral response to the potential aerial predator.Although both populations displayed a similar magnitude of response,they differed in the specific behaviors used. Upstream fishtended to freeze on the tank bottom, whereas downstream fishtended to hide under shelter. Field observations suggest thatthese strategies are related to habitat features specific toeach site. The behaviors used against aerial predators differedsubstantially from the behaviors used against aquatic predators,suggesting that different types of predators exert conflictingselection pressures. This research demonstrates the importanceof considering multiple selection pressures acting on an organismwhen trying to understand the evolutionary history of behavioraland morphological traits.     

Processive kinesins require loose mechanical coupling for efficient collective motility

Processive motor proteins are stochastic steppers that perform actual mechanical steps for only a minor fraction of the time they are bound to the filament track. Motors usually work in teams and therefore the question arises whether the stochasticity of stepping can cause mutual interference when motors are mechanically coupled. We used biocompatible surfaces to immobilize processive kinesin-1 motors at controlled surface densities in a mechanically well-defined way. This helped us to study quantitatively how mechanical coupling between motors affects the efficiency of collective microtubule transport. We found that kinesin-1 constructs that lack most of the non-motor sequence slow each other down when collectively transporting a microtubule, depending on the number of interacting motors. This negative interference observed for a motor ensemble can be explained quantitatively by a mathematical model using the known physical properties of individual molecules of kinesin-1. The non-motor extension of kinesin-1 reduces this mutual interference, indicating that loose mechanical coupling between motors is required for efficient transport by ensembles of processive motors.     

Simultaneous production of β-carotene, vitamin E and vitamin C by the aerial microalga Trentepohlia aurea

The biomass production and antioxidant accumulation of the aerial microalga Trentepohlia aurea were examined. Using ammonium chloride as nitrogen source for growth in liquid medium, the growth rate was exponential and accompanied by a marked fall in the pH of the medium. The highest growth rate of 152 mg L^-1 day^-1 (logarithmic phase) was attained when T. aurea was cultured with aeration of 5% CO₂-enriched air in medium buffered with HEPES. The growth rate and antioxidant content were enhanced under 5% CO₂ by switching the light intensity from 43 to 143 μmol photon m^-2 s^-1 for the two-stage culture. As a result, T. aurea cells accumulated 2.1 mg β-carotene, 0.3 mg L-ascorbic acid and 2.4 mg tocopherols, respectively, per g dry cell. The simultaneous production of useful materials, such as β-carotene, vitamin E and vitamin C, was demonstrated. This revised version was published online in June 2006 with corrections to the Cover Date.     

Contribution of aerial hyphae of Aspergillus oryzae to respiration in a model solid-state fermentation system

Oxygen transfer is for two reasons a major concern in scale-up and process control in industrial application of aerobic fungal solid-state fermentation (SSF): 1) heat production is proportional to oxygen uptake and it is well known that heat removal is one of the main problems in scaled-up fermenters, and 2) oxygen supply to the mycelium on the surface of or inside the substrate particles may be hampered by diffusion limitation. This article gives the first experimental evidence that aerial hyphae are important for fungal respiration in SSF. In cultures of A. oryzae on a wheat-flour model substrate, aerial hyphae contributed up to 75% of the oxygen uptake rate by the fungus. This is due to the fact that A. oryzae forms very abundant aerial mycelium and diffusion of oxygen in the gas-filled pores of the aerial hyphae layer is rapid. It means that diffusion limitation in the densely packed mycelium layer that is formed closer to the substrate surface and that has liquid-filled pores is much less important for A. oryzae than was previously reported for R. oligosporus and C. minitans. It also means that the overall oxygen uptake rate for A. oryzae is much higher than the oxygen uptake rate that can be predicted in the densely packed mycelium layer for R. oligosporus and C. minitans. This would imply that cooling problems become more pronounced. Therefore, it is very important to clarify the physiological role of aerial hyphae in SSF.