Investigation of two new putative pheromone components of the invasive Australian redback spider,Latrodectus hasseltii,with potential applications for control

The Australian redback spider, Latrodectus hasseltii preys on at least 10 endemic species in New Zealand, highlighting a need for control. Male redbacks are attracted to virgin females by an airborne pheromone. The aim of this study was to analyse the response of male redback spiders to two volatile chemicals found on the silk of virgin but not mated females, to determine whether these compounds constitute components of the airborne pheromone. Mature male redback spiders were placed in an olfactometer where they had a choice of two stimuli. We compared their response to paired combinations of a control, virgin silk, butyric acid and isovaleric acid. Male redbacks responded equally strongly to butyric acid and virgin silk, in terms of time spent near the stimulus. The identification of butyric acid as a component of the airborne sex pheromone of L. hasseltii provides the groundwork for developing a pheromone-based control.     

Using continuous directed evolution to improve enzymes for plant applications

Continuous directed evolution of enzymes and other proteins in microbial hosts is capable of outperforming classical directed evolution by executing hypermutation and selection concurrently in vivo, at scale, with minimal manual input. Provided that a target enzyme’s activity can be coupled to growth of the host cells, the activity can be improved simply by selecting for growth. Like all directed evolution, the continuous version requires no prior mechanistic knowledge of the target. Continuous directed evolution is thus a powerful way to modify plant or non-plant enzymes for use in plant metabolic research and engineering. Here, we first describe the basic features of the yeast (Saccharomyces cerevisiae) OrthoRep system for continuous directed evolution and compare it briefly with other systems. We then give a step-by-step account of three ways in which OrthoRep can be deployed to evolve primary metabolic enzymes, using a THI4 thiazole synthase as an example and illustrating the mutational outcomes obtained. We close by outlining applications of OrthoRep that serve growing demands (i) to change the characteristics of plant enzymes destined for return to plants, and (ii) to adapt (“plantize”) enzymes from prokaryotes—especially exotic prokaryotes—to function well in mild, plant-like conditions.

Continuous directed evolution using the yeast OrthoRep system is a powerful way to improve enzymes for use in plant engineering as illustrated by “plantizing” a bacterial thiamin synthesis enzyme.     

A framework to evaluate whether to pool or separate behaviors in a multilayer network

A multilayer network approach combines different network layers,which are connected by interlayer edges,to create a single mathematical object.These networks can contain a variety of information types and represent different aspects of a system.However,the process for selecting which information to include is not always straightforward.Using data on 2 agonistic behaviors in a captive population of monk parakeets(Myiopsitta monachus),we developed a framework for investigating how pooling or splitting behaviors at the scale of dyadic relationships(between 2 individuals)affects individual-and group-level social properties.We designed 2 reference models to test whether randomizing the number of interactions across behavior types results in similar structural patterns as the observed data.Although the behaviors were correlated,the first reference model suggests that the 2 behaviors convey different information about some social properties and should therefore not be pooled.However,once we controlled for data sparsity,we found that the observed measures corresponded with those from the second reference model.Hence,our initial result may have been due to the unequal frequencies of each behavior.Overall,our findings support pooling the 2 behaviors.Awareness of how selected measurements can be affected by data properties is warranted,but nonetheless our framework disentangles these efforts and as a result can be used for myriad types of behaviors and questions.This framework will help researchers make informed and data-driven decisions about which behaviors to pool or separate,prior to using the data in subsequent multilayer network analyses.     

Effect of increased gas density on pulmonary gas exchange in man.

Pulmonary gas exchange was measured in seven resting supine subjects breathing air or a dense gas mixture containing 21% O2 in sulfur hexafluoride (SF6). The mean value of the alveolar-arterial oxygen difference (AaDO2) decreased from 12.4 on air to 7.0 on SF6 (P less than 0.01), and increased again to 13.4 when air breathing resumed (P less than 0.01). No differences occurred between gas mixtures for O2 consumption, respiratory quotient, minute ventilation, breathing frequency, heart rate, or blood pressure, and the improved oxygen transfer could not be attributed to changes in cardiac output or mixed venous oxygen content in the one subject in which they were measured. These results are best explained by an altered distribution of ventilation during dense gas breathing, so that the ventilation-perfusion ratio (VA/Q) variance was reduced. Of several considered mechanisms, we favor one in which SF6 promotes cardiogenic gas mixing between peripheral parallel units having different alveolar gas concentrations. This mechanism allows for observed increases in arterial carbon dioxide tension and dead space-to-tidal volume ratio during dense gas breathing, and suggests that intraregional VA/Q variance accounts for at least one-half of the resting AaDO2 in healthy supine young men.     

Spatial ecology of wolverines at the southern periphery of distribution

Wolverines (Gulo gulo) in the conterminous United States have experienced range contraction, are uncommon, and have been designated as warranted for protection under the United States Endangered Species Act. Data from the southern edge of the wolverine's circumpolar distribution is sparse, and development of effective conservation strategies would benefit from a more complete understanding of the species' ecology. We captured and radio-monitored 30 wolverines in the Greater Yellowstone Ecosystem (GYE), tested for seasonal habitat selection by elevation band, and examined a suite of spatial characteristics to clarify our understanding of the wolverine's niche. Wolverines in GYE selected for areas >2,600 m latitude-adjusted elevation (LAE; n = 2,257 wolverine locations [12 F, 6 M]). Wolverines avoided areas <2,150 m LAE, including during winter when the vast majority of ungulates are pushed to these elevations by deep snow. Wolverine home ranges were large relative to body size, averaging 303 km² for adult females and 797 km² for adult males (n = 13 [8 F, 5 M] and 33 wolverine-years). Resident adults fit with Global Positioning System (GPS) collars used an area >75% the size of their multi-year home range in an average of 32 days (n = 7 [5 F, 2 M]). Average movement rates of 1.3 km/2-hr indicated that both sexes move distances equivalent to the diameter of their home range every 2 days or the circumference of their home range in <1 week (n = 1,329 2-hr movements, n = 12 individuals [7 F, 5 M]). This capability for movement, the short time-frame over which home ranges were developed, and a lack of home range overlap by same sex adults ( , 90% CI = 0.0–4.8%, n = 22 pairs) suggested territoriality. We estimated wolverine density to be 3.5/1,000 km² of area >2,150 m LAE (95% CI = 2.8–9.6). Dispersal movements extended to at least 170 km for both sexes (n = 5 F, 2 M). At the southern edge of distribution, where suitable and unsuitable conditions exist in close proximity, wolverines selected high-elevation areas near alpine tree-line where a mix of forest, meadow, and boulder fields were present, deep snow-cover existed during winter, and low temperatures near freezing can occur throughout the year. Persistence in these areas where the growing season is brief requires large home ranges that are regularly patrolled, a social system that provides exclusive access to resources, and low densities. These characteristics, along with low reproductive rates, are prevalent throughout the species range, indicating that wolverines are specialists at exploiting a cold, unproductive niche where interspecific competition is limited. The vulnerability inherent in occupying this unproductive niche was likely influential in previous declines within the conterminous United States and will remain a factor as wolverines encounter modern human influences. Conserving wolverines in the conterminous United States will require collaborative management over a large geographic scale. © 2011 The Wildlife Society.     

Clostridium difficile clinical isolates exhibit variable susceptibility and proteome alterations upon exposure to mammalian cationic antimicrobial peptides

Clostridium difficile is a leading cause of hospital-acquired bacterial infections in the United States, and the increased incidence of recurrent C. difficile infections is particularly problematic. The molecular mechanisms of C. difficile colonization, including its ability to evade host innate immune responses, is poorly understood. We hypothesized that epidemic-associated C. difficile clinical isolates would exhibit increased resistance to mammalian, gut-associated, cationic antimicrobial peptides such as the cathelicidin LL-37. Standardized susceptibility tests as well as comparative proteomic analyses revealed that C. difficile strains varied in their responses to LL-37, with epidemic-associated 027 ribotype isolates displaying greater resistance. Further, exposure of C. difficile strains to sub-lethal concentrations of LL-37 resulted in increased resistance to subsequent peptide challenge, suggesting the presence of inducible resistance mechanisms. Correspondingly, LL-37 exposure altered the C. difficile proteome, with marked changes in abundance of cell wall biosynthesis proteins, surface layer proteins, ABC transporters and lysine metabolism pathway components. Taken together, these results suggest that innate immune avoidance mechanisms could facilitate robust colonization by C. difficile.     

Phylogenetic community structure during succession: Evidence from three Neotropical forest sites

The phylogenetic structure of communities can reveal forces shaping community assembly, but the vast majority of work on phylogenetic community structure has been conducted in mature ecosystems. Here, we present an analysis of the phylogenetic structure of three Neotropical rain forest communities undergoing succession. In each site, the net relatedness of the community is initially high and consistently declines during succession. This pattern is evident both when comparing plots of different age classes and when comparing stem size classes within each plot: the oldest plots and the youngest stem cohorts, representing the most advanced stages of succession, have the lowest relatedness. Our results suggest that succession leaves a distinct signature in the phylogenetic structure of communities, which may reflect an increasing role of biotic interactions in community assembly during succession. We discuss theoretical explanations for the decline in community phylogenetic relatedness during succession, and suggest directions for future study.