Drivers of intrapopulation variation in resource use in a generalist predator, the macaroni penguin

Intrapopulation variation in resource use occurs in many populations of generalist predators with important community and evolutionary implications. One of the hypothesised mechanisms for such widespread variation is ecological opportunity, i.e. resource availability determined by intrinsic constraints and extrinsic conditions. We combined tracking data and stable isotope analysis to examine how breeding constraints and prey conditions influenced intrapopulation variation in resource use among macaroni penguins Eudyptes chrysolophus. Isotopic variation was also examined as a function of breeding success, individual traits and individual specialisation. Variation in isotope ratios was greatest across multiple tissue types when birds were able to undertake mid-range foraging trips (i.e. during incubation and pre-moult). This variation was highly consistent between years that spanned a 3-fold difference in local krill Euphausia superba density and was also highly consistent at the individual level between 2 years that had similar krill densities. However, by comparing our results with previous work on the same population, it appeared that a decrease in local prey availability can increase intrapopulation variation in resource use during periods with more restricted foraging ranges (i.e. during brood-guard and crèche). This study highlights the importance of considering ecological interactions that operate on multiple spatio-temporal scales when examining the drivers of resource use in populations of generalist predators

Interplay of Staphylococcal and Host Proteases Promotes Skin Barrier Disruption in Netherton Syndrome.

Netherton syndrome (NS) is a monogenic skin disease resulting from loss of function of lymphoepithelial Kazal-type-related protease inhibitor (LEKTI-1). In this study we examine if bacteria residing on the skin are influenced by the loss of LEKTI-1 and if interaction between this human gene and resident bacteria contributes to skin disease. Shotgun sequencing of the skin microbiome demonstrates that lesional skin of NS subjects is dominated by Staphylococcus aureus (S. aureus) and Staphylococcus epidermidis (S. epidermidis). Isolates of either species from NS subjects are able to induce skin inflammation and barrier damage on mice. These microbes promote skin inflammation in the setting of LEKTI-1 deficiency due to excess proteolytic activity promoted by S. aureus phenol-soluble modulin α as well as increased bacterial proteases staphopain A and B from S. aureus or EcpA from S. epidermidis. These findings demonstrate the critical need for maintaining homeostasis of host and microbial proteases to prevent a human skin disease

Stochastic Expression of Sae-Dependent Virulence Genes during Staphylococcus aureus Biofilm Development Is Dependent on SaeS

The intricate process of biofilm formation in the human pathogen Staphylococcus aureus involves distinct stages during which a complex mixture of matrix molecules is produced and modified throughout the developmental cycle. Early in biofilm development, a subpopulation of cells detaches from its substrate in an event termed “exodus” that is mediated by SaePQRS-dependent stochastic expression of a secreted staphylococcal nuclease, which degrades extracellular DNA within the matrix, causing the release of cells and subsequently allowing for the formation of metabolically heterogenous microcolonies. Since the SaePQRS regulatory system is involved in the transcriptional control of multiple S. aureus virulence factors, the expression of several additional virulence genes was examined within a developing biofilm by introducing fluorescent gene reporter plasmids into wild-type S. aureus and isogenic regulatory mutants and growing these strains in a microfluidic system that supplies the bacteria with a constant flow of media while simultaneously imaging developing biofilms in 5-min intervals. This study demonstrated that multiple virulence genes, including nuc, were expressed stochastically within a specialized subpopulation of cells in nascent biofilms. We demonstrated that virulence genes regulated by SaePQRS were stochastically expressed in nearly all strains examined whereas Agr-regulated genes were expressed more homogenously within maturing microcolonies. The commonly used Newman strain contains a variant of SaeS (SaeSP) that confers constitutive kinase activity to the protein and caused this strain to lack the stochastic expression pattern observed in other strain backgrounds. Importantly, repair of the SaeSP allele resulting in reversion to the well-conserved SaeSL allele found in other strains restored stochastic expression in this strain

Eye movements and hazard perception in active and passive driving

Differences in eye movement patterns are often found when comparing passive viewing paradigms to actively engaging in everyday tasks. Arguably, investigations into visuomotor control should therefore be most useful when conducted in settings that incorporate the intrinsic link between vision and action. We present a study that compares oculomotor behaviour and hazard reaction times across a simulated driving task and a comparable, but passive, video-based hazard perception task. We found that participants scanned the road less during the active driving task and fixated closer to the front of the vehicle. Participants were also slower to detect the hazards in the driving task. Our results suggest that the interactivity of simulated driving places increased demand upon the visual and attention systems than simply viewing driving movies. We offer insights into why these differences occur and explore the possible implications of such findings within the wider context of driver training and assessment

Low levels of β-lactam antibiotics induce extracellular DNA release and biofilm formation in Staphylococcus aureus.

UNLABELLED: Subminimal inhibitory concentrations of antibiotics have been shown to induce bacterial biofilm formation. Few studies have investigated antibiotic-induced biofilm formation in Staphylococcus aureus, an important human pathogen. Our goal was to measure S. aureus biofilm formation in the presence of low levels of β-lactam antibiotics. Fifteen phylogenetically diverse methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) strains were employed. Methicillin, ampicillin, amoxicillin, and cloxacillin were added to cultures at concentrations ranging from 0× to 1× MIC. Biofilm formation was measured in 96-well microtiter plates using a crystal violet binding assay. Autoaggregation was measured using a visual test tube settling assay. Extracellular DNA was quantitated using agarose gel electrophoresis. All four antibiotics induced biofilm formation in some strains. The amount of biofilm induction was as high as 10-fold and was inversely proportional to the amount of biofilm produced by the strain in the absence of antibiotics. MRSA strains of lineages USA300, USA400, and USA500 exhibited the highest levels of methicillin-induced biofilm induction. Biofilm formation induced by low-level methicillin was inhibited by DNase. Low-level methicillin also induced DNase-sensitive autoaggregation and extracellular DNA release. The biofilm induction phenotype was absent in a strain deficient in autolysin (atl). Our findings demonstrate that subminimal inhibitory concentrations of β-lactam antibiotics significantly induce autolysin-dependent extracellular DNA release and biofilm formation in some strains of S. aureus. IMPORTANCE: The widespread use of antibiotics as growth promoters in agriculture may expose bacteria to low levels of the drugs. The aim of this study was to investigate the effects of low levels of antibiotics on bacterial autoaggregation and biofilm formation, two processes that have been shown to foster genetic exchange and antibiotic resistance. We found that low levels of β-lactam antibiotics, a class commonly used in both clinical and agricultural settings, caused significant autoaggregation and biofilm formation by the important human pathogen Staphylococcus aureus. Both processes were dependent on cell lysis and release of DNA into the environment. The effect was most pronounced among multidrug-resistant strains known as methicillin-resistant S. aureus (MRSA). These results may shed light on the recalcitrance of some bacterial infections to antibiotic treatment in clinical settings and the evolution of antibiotic-resistant bacteria in agricultural settings

Minimising the limitations of using dietary analysis to assess foodweb changes by combining multiple techniques

Dietary studies of marine predators offer an immediate signal of foodweb changes occurring at lower trophic levels, and therefore are often used to assess the ecosystem status of marine systems. Conventionally, these studies are based on morphological analysis of prey remains in stomach contents, involving invasive and destructive techniques to collect samples. More recently, the number of dietary studies based on less invasive biochemical and molecular approaches has dramatically increased. However, all three methods, morphological, biochemical and molecular, have well-documented limitations for resolving taxonomy, temporal variation or biomass composition. In this study, we minimise these limitations by considering multiple techniques in combination. As a case study, we report the target prey species and diet composition of a marine predator that has been used to assess annual change in managed fishing areas for several decades, the macaroni penguin Eudyptes chrysolophus. We use biochemical (stable isotope) and molecular (DNA) analysis of faecal samples collected across the different phases of a single breeding season, and compare the resolved diet to a 26-year dataset of stomach contents collected from a closely located colony (0.25 km apart) that exploits identical foraging grounds. Molecular analysis increased the known target prey species for this highly monitored population by 31%, including a fish species of commercial importance. Biochemical analysis detected subtle changes in the proportion of fish and krill in the diet, demonstrating promising opportunities for using a combined molecular and biochemical method to assess inter-annual foodweb changes at lower trophic levels. The combined approach offers a less invasive sampling methodology, compared to morphological analysis, and provides more information regarding prey species diversity and the overall trophic signature of the diet. Further studies are required to examine the feasibility of using this approach for long-term dietary studies of different marine predator species

The sensitivity of seabird populations to density-dependence, environmental stochasticity and anthropogenic mortality

The balance between economic growth and wildlife conservation is a priority for many governments. Enhancing realism in assessment of population‐level impacts of anthropogenic mortality can help achieve this balance. Population Viability Analysis (PVA) is commonly applied to investigate population vulnerability, but outcomes of PVA are sensitive to formulations of density‐dependence, environmental stochasticity and life history. Current practice in marine assessments is to use precautionary models that assume no compensation from density‐dependence or rescue‐effects via “re‐seeding” from other colonies. However, if we could empirically quantify regulatory population processes, the responses of populations to additional anthropogenic mortality may be assessed with more realism in PVA. Using Bayesian state‐space models fitted to population time series from three sympatric seabird populations, selected for varied life histories, we inferred the extent to which their dynamics are driven by environmental stochasticity and density‐dependence. Based on these inferences, we conducted an exhaustive PVA across credible parameterizations for intrinsic and extrinsic population regulation, simulated as a closed and re‐seeded system. Scenarios of anthropogenic mortality, along a sliding scale of precaution, were applied both proportionally and as a fixed quota using Potential Biological Removal (PBR). Baseline results from fitting revealed clear environmental regulation in two of our three species. Crucially, we found that for our empirically derived, realistic model parameterizations there are risks of decline to real populations even under very precautionary mortality scenarios. We find that PBR is dubious in application as a sustainable tool for population assessment when we account for regulation. Closed versus re‐seeded models showed a large divergence in outcomes, with sharper declines in closed simulations. Fixed‐quota mortality typically induced greater population declines comparative to proportional mortality, subject to regulation and re‐seeding. Synthesis and applications. Practitioners using arbitrary formulations of population regulation risk over‐precaution (economic constraint) or under‐precaution (endangering populations). The demands of increased economic development and preservation of wildlife require that methodologies apply techniques that confer reality and rigour to assessment. The current practice of employing models lacking density‐dependence and empirical environmental information imposes limitations in the efficacy of estimating impacts. Here, we provide a method to quantify the conditions that predominantly regulate a population and exacerbate the risk of decline from anthropogenic mortality. It is in the interests of both developers and conservationists to apply methods in population impact assessments that capture realism in the processes driving population dynamics

Identification of Extracellular DNA-Binding Proteins in the Biofilm Matrix.

We developed a new approach that couples Southwestern blotting and mass spectrometry to discover proteins that bind extracellular DNA (eDNA) in bacterial biofilms. Using Staphylococcus aureus as a model pathogen, we identified proteins with known DNA-binding activity and uncovered a series of lipoproteins with previously unrecognized DNA-binding activity. We demonstrated that expression of these lipoproteins results in an eDNA-dependent biofilm enhancement. Additionally, we found that while deletion of lipoproteins had a minimal impact on biofilm accumulation, these lipoprotein mutations increased biofilm porosity, suggesting that lipoproteins and their associated interactions contribute to biofilm structure. For one of the lipoproteins, SaeP, we showed that the biofilm phenotype requires the lipoprotein to be anchored to the outside of the cellular membrane, and we further showed that increased SaeP expression correlates with more retention of high-molecular-weight DNA on the bacterial cell surface. SaeP is a known auxiliary protein of the SaeRS system, and we also demonstrated that the levels of SaeP correlate with nuclease production, which can further impact biofilm development. It has been reported that S. aureus biofilms are stabilized by positively charged cytoplasmic proteins that are released into the extracellular environment, where they make favorable electrostatic interactions with the negatively charged cell surface and eDNA. In this work we extend this electrostatic net model to include secreted eDNA-binding proteins and membrane-attached lipoproteins that can function as anchor points between eDNA in the biofilm matrix and the bacterial cell surface.IMPORTANCE Many bacteria are capable of forming biofilms encased in a matrix of self-produced extracellular polymeric substances (EPS) that protects them from chemotherapies and the host defenses. As a result of these inherent resistance mechanisms, bacterial biofilms are extremely difficult to eradicate and are associated with chronic wounds, orthopedic and surgical wound infections, and invasive infections, such as infective endocarditis and osteomyelitis. It is therefore important to understand the nature of the interactions between the bacterial cell surface and EPS that stabilize biofilms. Extracellular DNA (eDNA) has been recognized as an EPS constituent for many bacterial species and has been shown to be important in promoting biofilm formation. Using Staphylococcus aureus biofilms, we show that membrane-attached lipoproteins can interact with the eDNA in the biofilm matrix and promote biofilm formation, which suggests that lipoproteins are potential targets for novel therapies aimed at disrupting bacterial biofilms

Coping with strangers: how familiarity and active interactions shape group coordination in Corydoras aeneus.

Social groups composed of familiar individuals exhibit better coordination than unfamiliar groups; however, the ways familiarity contributes to coordination are poorly understood. Prior social experience probably allows individuals to learn the tendencies of familiar group-mates and respond accordingly. Without prior experience, individuals would benefit from strategies for enhancing coordination with unfamiliar others. We used a social catfish, Corydoras aeneus, that uses discrete, observable tactile interactions to assess whether active interactions could facilitate coordination, and how their role might be mediated by familiarity. We describe this previously understudied physical interaction, 'nudges', and show it to be associated with group coordination and cohesion. Furthermore, we investigated nudging and coordination in familiar/unfamiliar pairs. In all pairs, we found that nudging rates were higher during coordinated movements than when fish were together but not coordinating. We observed no familiarity-based difference in coordination or cohesion. Instead, unfamiliar pairs exhibited significantly higher nudging rates, suggesting that unfamiliar pairs may be able to compensate for unfamiliarity through increased nudging. By contrast, familiar individuals coordinated with comparatively little nudging. Second, we analysed nudging and cohesion within triplets of two familiar and one unfamiliar individual (where familiar individuals had a choice of partner). Although all individuals nudged at similar rates, the unfamiliar group-mate was less cohesive than its familiar group-mates and spent more time alone. Unfamiliar individuals that nudged their group-mates more frequently exhibited higher cohesion, indicating that nudging may facilitate cohesion for the unfamiliar group-mate. Overall, our results suggest that nudges can mitigate unfamiliarity, but that their usage is reduced in the case of familiar individuals, implying a cost is associated with the behaviour