Plant phylogeny as a surrogate for turnover in beetle assemblages

The ability to extrapolate from the known to the unknown is essential if we are to use the turnover of overall biodiversity, as opposed to a few well-known groups, to inform conservation planning. We investigated the usefulness of using evolutionary relationships of plants as a surrogate for the turnover of their associated beetle assemblages. If plant traits that are important to insects are phylogenetically conserved, it follows that there will be a positive relationship between insect faunal dissimilarity and plant evolutionary distance. We collected beetles using pyrethrum knock-down methods from 40 plant species belonging to four plant families in the Sydney region of Eastern Australia. We developed a novel approach for estimating variance in the dissimilarity of beetle assemblages, as explained by plant phylogeny, by using phylogenetic eigenvectors as explanatory variables in a distance-based redundancy analysis. We found a highly significant relationship between faunal dissimilarity and plant evolutionary distance for the entire beetle assemblage, the herbivorous component, and the non-herbivorous component, indicating that beetles generally showed some preference for particular plant clades as habitat, regardless of feeding guild. When comparing observed dissimilarities with those predicted from 40 jack-knife replicates of a Generalised Dissimilarity Model, we were often able to predict beetle turnover from plant phylogenetic relationships, although the reliability of this result was highly variable. Nevertheless, the broad response of beetle assemblages to plant evolutionary relatedness indicates real potential for plant phylogenetic pattern to act as a useful surrogate for insect biodiversity, especially when supplemented with other environmental correlates.     

Effect of Streptococcus salivarius K12 on the in vitro growth of Candida albicans and its protective effect in an oral candidiasis model

Oral candidiasis is often accompanied by severe inflammation, resulting in a decline in the quality of life of immunosuppressed individuals and elderly people. To develop a new oral therapeutic option for candidiasis, a nonpathogenic commensal oral probiotic microorganism, Streptococcus salivarius K12, was evaluated for its ability to modulate Candida albicans growth in vitro, and its therapeutic activity in an experimental oral candidiasis model was tested. In vitro inhibition of mycelial growth of C. albicans was determined by plate assay and fluorescence microscopy. Addition of S. salivarius K12 to modified RPMI 1640 culture medium inhibited the adherence of C. albicans to the plastic petri dish in a dose-dependent manner. Preculture of S. salivarius K12 potentiated its inhibitory activity for adherence of C. albicans. Interestingly, S. salivarius K12 was not directly fungicidal but appeared to inhibit Candida adhesion to the substratum by preferentially binding to hyphae rather than yeast. To determine the potentially anti-infective attributes of S. salivarius K12 in oral candidiasis, the probiotic was administered to mice with orally induced candidiasis. Oral treatment with S. salivarius K12 significantly protected the mice from severe candidiasis. These findings suggest that S. salivarius K12 may inhibit the process of invasion of C. albicans into mucous surfaces or its adhesion to denture acrylic resins by mechanisms not associated with the antimicrobial activity of the bacteriocin. S. salivarius K12 may be useful as a probiotic as a protective tool for oral care, especially with regard to candidiasis.     

Reliability of genetic bottleneck tests for detecting recent population declines

The identification of population bottlenecks is critical in conservation because populations that have experienced significant reductions in abundance are subject to a variety of genetic and demographic processes that can hasten extinction. Genetic bottleneck tests constitute an appealing and popular approach for determining if a population decline has occurred because they only require sampling at a single point in time, yet reflect demographic history over multiple generations. However, a review of the published literature indicates that, as typically applied, microsatellite-based bottleneck tests often do not detect bottlenecks in vertebrate populations known to have experienced declines. This observation was supported by simulations that revealed that bottleneck tests can have limited statistical power to detect bottlenecks largely as a result of limited sample sizes typically used in published studies. Moreover, commonly assumed values for mutation model parameters do not appear to encompass variation in microsatellite evolution observed in vertebrates and, on average, the proportion of multi-step mutations is underestimated by a factor of approximately two. As a result, bottleneck tests can have a higher probability of 'detecting' bottlenecks in stable populations than expected based on the nominal significance level. We provide recommendations that could add rigor to inferences drawn from future bottleneck tests and highlight new directions for the characterization of demographic history.     

An ecological approach to assessing the epidemiology of antimicrobial resistance in animal and human populations

We examined long-term surveillance data on antimicrobial resistance (AMR) in Salmonella Typhimurium DT104 (DT104) isolates from concurrently sampled and sympatric human and animal populations in Scotland. Using novel ecological and epidemiological approaches to examine diversity, and phenotypic and temporal relatedness of the resistance profiles, we assessed the more probable source of resistance of these two populations. The ecological diversity of AMR phenotypes was significantly greater in human isolates than in animal isolates, at the resolution of both sample and population. Of 5200 isolates, there were 65 resistance phenotypes, 13 unique to animals, 30 unique to humans and 22 were common to both. Of these 22, 11 were identified first in the human isolates, whereas only five were identified first in the animal isolates. We conclude that, while ecologically connected, animals and humans have distinguishable DT104 communities, differing in prevalence, linkage and diversity. Furthermore, we infer that the sympatric animal population is unlikely to be the major source of resistance diversity for humans. This suggests that current policy emphasis on restricting antimicrobial use in domestic animals may be overly simplistic. While these conclusions pertain to DT104 in Scotland, this approach could be applied to AMR in other bacteria-host ecosystems.     

Preferential induction of CD4+ T cell responses through in vivo targeting of antigen to dendritic cell-associated C-type lectin-1

Targeting of Ags and therapeutics to dendritic cells (DCs) has immense potential for immunotherapy and vaccination. Because DCs are heterogeneous, optimal targeting strategies will require knowledge about functional specialization among DC subpopulations and identification of molecules for targeting appropriate DCs. We characterized the expression of a fungal recognition receptor, DC-associated C-type lectin-1 (Dectin-1), on mouse DC subpopulations and investigated the ability of an anti-Dectin-1 Ab to deliver Ag for the stimulation of immune responses. Dectin-1 was shown to be expressed on CD8alpha-CD4-CD11b+ DCs found in spleen and lymph nodes and dermal DCs present in skin and s.c. lymph nodes. Injection of Ag-anti-Dectin-1 conjugates induced CD4+ and CD8+ T cell and Ab responses at low doses where free Ag failed to elicit a response. Notably, qualitatively different immune responses were generated by targeting Ag to Dectin-1 vs CD205, a molecule expressed on CD8alpha+CD4-CD11b- DCs, dermal DCs, and Langerhans cells. Unlike anti-Dectin-1, anti-CD205 conjugates failed to elicit an Ab response. Moreover, when conjugates were injected i.v., anti-Dectin-1 stimulated a much stronger CD4+ T cell response and a much weaker CD8+ T cell response than anti-CD205. The results reveal Dectin-1 as a potential targeting molecule for immunization and have implications for the specialization of DC subpopulations.     

In vivo rates of erythrocyte glutathione synthesis in adults with sickle cell disease

Despite reports of lower GSH concentration in sickle cell disease (SCD), the in vivo kinetic mechanism(s) responsible for GSH deficiency is unknown. To determine whether suppressed synthesis was responsible for the lower erythrocyte GSH concentration, we used a primed intermittent infusion of [(2)H(2)]glycine to measure erythrocyte GSH synthesis in vivo in 23 individuals with homozygous beta(s) SCD and 8 healthy controls. Erythrocyte cysteine concentration, the rate-limiting precursor for GSH synthesis, plasma markers of oxidant damage, and dietary intakes of energy and protein were also measured. Compared with values of controls, SCD subjects had significantly lower erythrocyte GSH (P < 0.04) and cysteine concentrations (P < 0.004) but significantly faster fractional rates of GSH synthesis (P < 0.02). The absolute rates of GSH synthesis in SCD subjects compared with control subjects was greater by approximately 57% (P = 0.062). However, the concentrations of markers of oxidative damage, plasma derivatives of reactive oxygen metabolites, plasma nitrotyrosine, urinary isoprostane-to-creatinine ratio, and GSH-to-GSSG ratio, as well as dietary intakes of energy, protein, and GSH precursor amino acids, were not different between SCD subjects and controls. The findings of this study suggest that the lower erythrocyte GSH of SCD patients is not due to suppressed synthesis or impaired regeneration but rather to increased consumption. In addition, the lower erythrocyte cysteine concentration plus the faster rate of GSH synthesis strongly suggest that the endogenous cysteine supply is not sufficient to meet all anabolic demands; hence, cysteine may be a conditionally essential amino acid in individuals with SCD.     

Megaplasmids encode differing combinations of lantibiotics in Streptococcus salivarius

Streptococcus salivarius strains commonly produce bacteriocins as putative anticompetitor or signalling molecules. Here we report that bacteriocin production by the oral probiotic strain S. salivarius K12 is encoded by a large (ca. 190 kb) plasmid. Oral cavity transmission of the plasmid from strain K12 to a plasmid-negative variant of this bacterium was demonstrated in two subjects. Tests of additional S. salivarius strains showed large (up to ca. 220 kb) plasmids present in bacteriocin-producing isolates. Various combinations (up to 3 per plasmid) of loci encoding the known streptococcal lantibiotics salivaricin A, salivaricin B, streptin and SA-FF22 were localised to these plasmids. Since all bacteriocin-producing strains of S. salivarius tested to date appear to harbour plasmids, it appears that they may function as mobile repositories for bacteriocin loci, especially those of the lantibiotic class.     

Macroinvertebrate abundance, water chemistry, and wetland characteristics affect use of wetlands by avian species in Maine

Our objective was to determine use by avian species (e.g., piscivores, marsh birds, waterfowl, selected passerines) of 29 wetlands in areas with low (<200 μeq l⁻¹) acid-neutralizing capacity (ANC) in southeastern Maine. We documented bird, pair, and brood use during 1982–1984 and in 1982 we sampled 10 wetlands with a sweep net to collect invertebrates. We related mean numbers of invertebrates per wetland to water chemistry, basin characteristics, and avian use of different wetland types. Shallow, beaver (Castor canadensis)-created wetlands with the highest phosphorus levels and abundant and varied macrophyte assemblages supported greater densities of macroinvertebrates and numbers of duck broods (88.3% of all broods) in contrast to deep, glacial type wetlands with sparse vegetation and lower invertebrate densities that supported fewer broods (11.7%). Low pH may have affected some acid-intolerant invertebrate taxa (i.e., Ephemeroptera), but high mean numbers of Insecta per wetland were recorded from wetlands with a pH of 5.51. Other Classes and Orders of invertebrates were more abundant on wetlands with pH > 5.51. All years combined use of wetlands by broods was greater on wetlands with pH ≤ 5.51 (77.4%) in contract to wetlands with pH > 5.51 that supported 21.8% of the broods. High mean brood density was associated with mean number of Insecta per wetland. For lentic wetlands created by beaver, those habitats contained vegetative structure and nutrients necessary to provide cover to support invertebrate populations that are prey of omnivore and insectivore species. The fishless status of a few wetlands may have affected use by some waterfowl species and obligate piscivores.     

Spatial variation in demography and population growth rate: the importance of natal location

1. Understanding the pattern and magnitude of spatial variation in demography and population growth rate (lambda) is key to understanding the structure and dynamics of natural populations. However, such spatial variation is challenging to quantify. We use>20 years of individual life-history data to quantify small- and large-scale spatial variation in demography and lambda within a single population of red-billed choughs Pyrrhocorax pyrrhocorax on Islay, Scotland. Critically, we demonstrate a major importance of an individual's natal rather than current location in driving observed spatial variation. 2. Breeding success (the number of offspring fledged per breeding attempt) varied among individual chough nest sites but did not vary on a larger spatial scale across Islay. 3. The proportion of fledglings observed to survive to recruiting age varied markedly among individual nest sites and also varied more widely across Islay. Spatial capture-mark-recapture models defined two discrete geographical regions where fledgling survival differed significantly: choughs fledged in region 'BGE' were more likely to survive than choughs fledged in region 'CNSW' as both subadults and adults. 4. The asymptotic lambda attributable to breeding attempts in region BGE exceeded unity, and exceeded that attributable to breeding attempts in region CNSW. Relatively productive and unproductive regions therefore exist within this population. 5. Spatial variation in adult survival was better explained by an individual's natal region than the region where that individual settled to breed. Spatial variation in lambda would consequently have remained undetected had survival been measured across resident breeders rather than across individuals fledged in each region. Furthermore, breeding success was a weak predictor of a nest site's estimated productivity of recruits. 6. We therefore describe marked spatial variation in demography and lambda within a single population of a territorial vertebrate, mediated partly by long-term links between an individual's natal location and its subsequent life-history. Life-long monitoring of individuals of known origin may therefore be necessary to identify accurately subpopulations of intrinsically high and low lambda.     

A proton delivery pathway in the soluble fumarate reductase from Shewanella frigidimarina

The mechanism for fumarate reduction by the soluble fumarate reductase from Shewanella frigidimarina involves hydride transfer from FAD and proton transfer from the active-site acid, Arg-402. It has been proposed that Arg-402 forms part of a proton transfer pathway that also involves Glu-378 and Arg-381 but, unusually, does not involve any bound water molecules. To gain further insight into the importance of this proton pathway we have perturbed it by substituting Arg-381 by lysine and methionine and Glu-378 by aspartate. Although all the mutant enzymes retain measurable activities, there are orders-of-magnitude decreases in their k(cat) values compared with the wild-type enzyme. Solvent kinetic isotope effects show that proton transfer is rate-limiting in the wild-type and mutant enzymes. Proton inventories indicate that the proton pathway involves multiple exchangeable groups. Fast scan protein-film voltammetric studies on wild-type and R381K enzymes show that the proton transfer pathway delivers one proton per catalytic cycle and is not required for transporting the other proton, which transfers as a hydride from the reduced, protonated FAD. The crystal structures of E378D and R381M mutant enzymes have been determined to 1.7 and 2.1 A resolution, respectively. They allow an examination of the structural changes that disturb proton transport. Taken together, the results indicate that Arg-381, Glu-378, and Arg-402 form a proton pathway that is completely conserved throughout the fumarate reductase/succinate dehydrogenase family of enzymes.