Evaluation of individual protein errors in silver-stained two-dimensional gels

The relationship between spot volume and variation for all protein spots observed on large format 2D gels when utilising silver stain technology and a model system based on mammalian NSO cell extracts is reported. By running multiple gels we have shown that the reproducibility of data generated in this way is dependent on individual protein spot volumes, which in turn are directly correlated with the coefficient of variation. The coefficients of variation across all observed protein spots were highest for low abundant proteins which are the primary contributors to process error, and lowest for more abundant proteins. Using the relationship between spot volume and coefficient of variation we show it is necessary to calculate variation for individual protein spot volumes. The inherent limitations of silver staining therefore mean that errors in individual protein spot volumes must be considered when assessing significant changes in protein spot volume and not global error. (C) 2003 Elsevier Science (USA). All rights reserved.

Metamorphosis of a scleractinian coral in response to microbial biofilms

Microorganisms have been reported to induce settlement and metamorphosis in a wide range of marine invertebrate species. However, the primary cue reported for metamorphosis of coral larvae is calcareous coralline algae (CCA). Herein we report the community structure of developing coral reef biofilms and the potential role they play in triggering the metamorphosis of a scleractinian coral. Two-week-old biofilms induced metamorphosis in less than 10% of larvae, whereas metamorphosis increased significantly on older biofilms, with a maximum of 41% occurring on 8-week-old microbial films. There was a significant influence of depth in 4- and 8-week biofilms, with greater levels of metamorphosis occurring in response to shallow-water communities. Importantly, larvae were found to settle and metamorphose in response to microbial biofilms lacking CCA from both shallow and deep treatments, indicating that microorganisms not associated with CCA may play a significant role in coral metamorphosis. A polyphasic approach consisting of scanning electron microscopy, fluorescence in situ hybridization (FISH), and denaturing gradient gel electrophoresis (DGGE) revealed that coral reef biofilms were comprised of complex bacterial and microalgal communities which were distinct at each depth and time. Principal-component analysis of FISH data showed that the Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, and Cytophaga-Flavobacterium of Bacteroidetes had the largest influence on overall community composition. A low abundance of Archaea was detected in almost all biofilms, providing the first report of Archaea associated with coral reef biofilms. No differences in the relative densities of each subdivision of Proteobacteria were observed between slides that induced larval metamorphosis and those that did not. Comparative cluster analysis of bacterial DGGE patterns also revealed that there were clear age and depth distinctions in biofilm community structure; however, no difference was detected in banding profiles between biofilms which induced larval metamorphosis and those where no metamorphosis occurred. This investigation demonstrates that complex microbial communities can induce coral metamorphosis in the absence of CCA.

Lineages of acidophilic archaea revealed by community genomic analysis

Novel, low-abundance microbial species can be easily overlooked in standard polymerase chain reaction (PCR)-based surveys. We used community genomic data obtained without PCR or cultivation to reconstruct DNA fragments bearing unusual 16S ribosomal RNA ( rRNA) and protein-coding genes from organisms belonging to novel archaeal lineages. The organisms are minor components of all biofilms growing in pH 0.5 to 1.5 solutions within the Richmond Mine, California. Probes specific for 16S rRNA showed that the fraction less than 0.45 micrometers in diameter is dominated by these organisms. Transmission electron microscope images revealed that the cells are pleomorphic with unusual folded membrane protrusions and have apparent volumes of < 0.006 cubic micrometer.

Do predators aggregate in response to pest density in agroecosystems? Assessing within-field spatial patterns

1. The spatial heterogeneity of predator populations is an important component of ecological theories pertaining to predator-prey dynamics. Most studies within agricultural fields show spatial correlation (positive or negative) between mean predator numbers and prey abundance across a whole field over time but generally ignore the within-field spatial dimension. We used explicit spatial mapping to determine if generalist predators aggregated within a soybean field, the size of these aggregations and if predator aggregation was associated with pest aggregation, plant damage and predation rate. 2. The study was conducted at Gatton in the Lockyer Valley, 90 km west of Brisbane, Australia. Intensive sampling grids were used to investigate within-field spatial patterns. The first row of each grid was located in a lucerne field (10 m from interface) and the remaining rows were in an adjacent soybean field. At each point on the grid the abundance of foliage-dwelling and ground-dwelling pests and predators was measured, predation rates [using sentinel Helicoverpa armigera (Hubner) egg cards] and plant damage were estimated. Eight grids were sampled across two summer cropping seasons (2000/01, 2001/02). 3. Predators exhibited strong spatial patterning with regions of high and low abundance and activity within what are considered to be uniform soybean fields. Ground-dwelling and foliage-dwelling predators were often aggregated in patches approximately 40 m across. 4. Lycosidae (wolf spiders) displayed aggregation and were consistently more abundant within the lucerne, with a decreasing trap catch with distance from the lucrene/soybean interface. This trend was consistent between subsequent grids in a single field and between fields. 5. The large amount of spatial variability in within-field arthropod abundance (pests and predators) and activity (egg predation and plant damage) indicates that whole field averages were misleading. This result has serious implications for sampling of arthropod abundance and pest management decision-making based on scouting data. 6. There was a great deal of temporal change in the significant spatial patterns observed within a field at each sampling time point during a single season. Predator and pest aggregations observed in these fields were generally not stable for the entire season. 7. Predator aggregation did not correlate consistently with pest aggregation, plant damage or predation rate. Spatial patterns in predator abundance were not associated consistently with any single parameter measured. The most consistent positive association was between foliage-dwelling predators and pests (significant in four of seven grids). Inferring associations between predators and prey based on an intensive one-off sampling grid is difficult, due to the temporal variability in the abundance of each group. 8. Synthesis and applications. This study demonstrated that generalist predator populations are rarely distributed randomly and field edges and adjacent crops can have an influence on within-field predator abundance. This must be considered when estimating arthropod (pest and predator) abundance from a set of samples taken at random locations within a field.

Purification and characterisation of two cancer cachexia factors

Cachexia is a wasting phenomenon that often accompanies malignant disease. Its manifestation is associated with shortened survival and reduced responsiveness to anti-tumour therapy and as yet there is no established, effective amelioratory treatment. The MAC 16 model of cancer cachexia has been shown by many studies to closely mirror the human condition. Thus, cachexia is mediated by the presence of a small, slow-growing solid tumour that is mainly resistant to chemotherapy. In addition, the condition is largely attributable to aberrations in metabolic processes, while weight loss due to anorexia is negligible. Cachexia induced by the MAC 16 tumour, has been shown to be mediated by the production of tumour-derived circulatory catabolic factors, and the further elucidation of the structure of these molecules contributes towards the main content of this report. Thus, a factor with in vitro lipid-mobilising activity has been purified from the MAC 16 tumour, and has been found to have similarities to tumour-derived lipolytic factors published to date. Further work demonstrated that this factor was also purifiable from the urine of a patient with pancreatic cancer, and that it was capable of inducing weight loss in non tumour-bearing mice. Sequence analysis of the homogeneous material revealed an identity to Zn-α-2-glycoprotein, the significance of which is discussed. An additional factor, first detected as a result of its specific reactivity with a monoclonal antibody produced by fusion of splenocytes from MAC 16 tumour-bearing mice with mouse BALB/c myeloma cells, was identified as a co-purificant during studies to isolate the lipolytic factor. Subsequent purification of this material to homogeneity resulted in the determination of 18 of the N-terminal amino acids and revealed the highly glycosylated nature of its structure. Thus, this material (P24) was found to have an apparent molecular mass of 24kD of which 2kD was due to protein, while the remainder (92%) was due to the presence of carbohydrate groups. Sequence analysis of the protein core of P24 revealed an identity with Streptococcal pre-absorbing antigen (PA-Ag) in 11 of the amino acids, and the significance of this is discussed. P24 was shown to induce muscle protein breakdown in vitro and to induce cachexia in vivo, as measured by the depletion of fat (29%) and muscle (14%) tissue in the absence of a reduction of food and water intake. Further studies revealed that the same material was purifiable from the urine of patients with pancreatic cancer and was found to be detectable in the urine of cancer patients with weight loss greater than l.Skg/month. Thus, cachexia induced by the MAC 16 tumour in mice and by malignant disease in humans may be induced by similar mediators. Attempts to isolate the gene for P24 using information provided by the N-terminal protein sequence were unsuccessful. This was probably due to the low abundance o[ the material, as determined by protein purification studies; and the nature of the amino acids of the N-terminal sequence, which conferred a high degree o[ degeneracy to the oligonucleotides designed for the polymerase chain reaction.

The amyloid precursor protein (APP) binds the PIKfyve complex and modulates its function

Phosphoinositides are important components of eukaryotic membranes that are required for multiple forms of membrane dynamics. Phosphoinositides are involved in defining membrane identity, mediate cell signalling and control membrane trafficking events. Due to their pivotal role in membrane dynamics, phosphoinositide de-regulation contributes to various human diseases. In this review, we will focus on the newly emerging regulation of the PIKfyve complex, a phosphoinositide kinase that converts the endosomal phosphatidylinositol-3-phosphate [PI(3)P] to phosphatidylinositol-3,5-bisphosphate [PI(3,5)P2)], a low abundance phosphoinositide of outstanding importance for neuronal integrity and function. Loss of PIKfyve function is well known to result in neurodegeneration in both mousemodels and human patients. Our recent work has surprisingly identified the amyloid precursor protein (APP), the central molecule in Alzheimer s disease aetiology, as a novel interaction partner of a subunit of the PIKfyve complex, Vac14. Furthermore, it has been shown that APP modulates PIKfyve function and PI(3,5)P2 dynamics, suggesting that the APP gene family functions as regulator of PI(3,5)P2 metabolism. The recent advances discussed in this review suggest a novel, unexpected, â-amyloid-independent mechanism for neurodegeneration in Alzheimer s disease.

The interactive effects of predators, resources, and disturbance on freshwater snail populations from the Everglades

The origins of population dynamics depend on interplay between abiotic and biotic factors; the relative importance of each changing across space and time. Predation is a central feature of ecological communities that removes individuals (consumption) and alters prey traits (non-consumptive). Resource quality mitigates non-consumptive predator effects by stimulating growth and reproduction. Disturbance resets predator-prey interactions by removing both. I integrate experiments, time-series analysis, and performance trials to examine the relative importance of these on the population dynamics of a snail species by studying a variety of their traits. A review of ninety-three published articles revealed that snail abundance was much less in the Everglades and similar ecosystems compared to all other freshwater ecosystems considered. Separating consumptive from non-consumptive (cues) predator effects at different phosphorous levels with an experiment determined that phosphorous stimulated, but predator cues inhibited snail growth (34% vs. 23%), activity (38% vs. 53%), and reproductive effort (99% vs. 90%) compared to controls. Cues induced taller shells and smaller openings and moved to refugia where they reduced periphyton by 8%. Consumptive predator effects were minor in comparison. In a reciprocal transplant cage experiment along a predator cue and phosphorous gradient created by a canal, snails grew 10% faster and produced 37% more eggs far from the canal (fewer cues) when fed phosphorous-enriched periphyton from near the canal. Time-series analysis at four sites and predator performance trials reveal that phosphorous-enriched regions support larger snail populations, seasonal drying removes snails at all sites, crayfish negatively affect populations in enriched regions, and molluscivorous fish consume snails in the wet season. Combining these studies reveals interplay between resources, predators, and seasonality that limit snail populations in the Everglades and lead to their low abundance compared to other freshwater ecosystems. Resource quality is emerging as the critical factor because improving resources profoundly improved growth and reproduction; seasonal drying and predation become important at times and places. This work contributes to the general understanding in ecology of the relative importance of different factors that structure populations and provides evidence that bolsters monitoring efforts to assess the Comprehensive Everglades Restoration Plan that show phosphorous enrichment is a major driver of ecosystem change.

A molecular and stable isotopic approach to investigate the importance of algal and detrital energy pathways in a freshwater marsh

The relative importance of algal and detrital energy pathways remains a central question in wetlands ecology. We used bulk stable isotope analysis and fatty acid composition to investigate the relative contributions of periphyton (algae) and floc (detritus) in a freshwater wetland with the goal of determining the inputs of these resource pools to lower trophic-level consumers. All animal samples revealed fatty acid markers indicative of both microbial (detrital) and algal origins, though the relative contributions varied among species. Vascular plant markers were in low abundance in most consumers. Detritivory is important for chironomids and amphipods, as demonstrated by the enhanced bacterial fatty acids present in both consumers, while algal resources, in the form of periphyton, likely support ephemeropteran larvae. Invertebrates such as amphipods and grass shrimp appear to be important resources for small omnivorous fish, while Poecilia latipinna appear to strongly use periphyton and Ephemeroptera larvae as food sources. Both P. latipinna and Lepomis spp. assimilated small amounts of vascular plant debris, possibly due to unintentional ingestion of floc while foraging for invertebrates and insect larvae. Physid snails, Haitia spp., were characterized by considerably different fatty acid compositions than other taxa examined, and likely play a unique role in Everglades’ food webs.

(Table 1) Grain size composition of surface sediments from the Barents Sea shelf

The surface layer of bottom sediments on the Barents Sea shelf has an irregular but generally very low abundance of diatoms. Tests of species belonging to present-day diatom flora were absent in nearly half of samples; their abundance was only a few shells per gram of dry sediment in 30% of the samples, it was up to 100 shells per gram in 9% of the samples, and was in thousands of shells per gram in only 13% of the samples. The lowest abundances of diatom shells were found in sediments of the eastern and northeastern parts of the sea owing to unfavorable sedimentation conditions and deficiency of dissolved silica in water. But distribution of diatom species on the surface of bottom sediments is strictly consistent with their present-day ranges. About 30% of the samples contained re-deposited Cretaceous and Paleogene diatoms indicating that bottom sediments have largely formed by scouring and re-deposition of underlying material.

(Table 2) Salinity, pH, alkalinity and major ion concentrations in ANDRILL core AND-2A pore water

The compositional record of the AND-2A drillcore is examined using petrological, sedimentological, volcanological and geochemical analysis of clasts, sediments and pore waters. Preliminary investigations of basement clasts (granitoids and metasediments) indicate both local and distal sources corresponding to variable ice-volume and ice-flow directions. Low abundance of sedimentary clasts (e.g., arkose, litharenite) suggests reduced contributions from sedimentary covers while intraclasts (e.g., diamictite, conglomerate) attest to intrabasinal reworking. Volcanic material includes pyroclasts (e.g., pumice, scoria), sediments and lava. Primary and reworked tephra layers occur within the Early Miocene interval (1093 to 640 metres below sea floor mbsf). The compositions of volcanic clasts reveal a diversity of alkaline types derived from the McMurdo Volcanic Group. Finer-grained sediments (e.g., sandstone, siltstone) show increases in biogenic silica and volcanic glass from 230 to 780 mbsf and higher proportions of terrigenous material c. 350 to 750 mbsf and below 970 mbsf. Basement clast assemblages suggest a dominant provenance from the Skelton Glacier - Darwin Glacier area and from the Ferrar Glacier - Koettlitz Glacier area. Provenance of sand grains is consistent with clast sources. Thirteen Geochemical Units are established based on compositional trends derived from continuous XRF scanning. High values of Fe and Ti indicate terrigenous and volcanic sources, whereas high Ca values signify either biogenic or diagenic sources. Highly alkaline and saline pore waters were produced by chemical exchange with glass at moderately elevated temperatures.

Acoustic and Magnetic Techniques for the Isolation and Analysis of Cells in Microfluidic Platforms

Cancer comprises a collection of diseases, all of which begin with abnormal tissue growth from various stimuli, including (but not limited to): heredity, genetic mutation, exposure to harmful substances, radiation as well as poor dieting and lack of exercise. The early detection of cancer is vital to providing life-saving, therapeutic intervention. However, current methods for detection (e.g., tissue biopsy, endoscopy and medical imaging) often suffer from low patient compliance and an elevated risk of complications in elderly patients. As such, many are looking to “liquid biopsies” for clues into presence and status of cancer due to its minimal invasiveness and ability to provide rich information about the native tumor. In such liquid biopsies, peripheral blood is drawn from patients and is screened for key biomarkers, chiefly circulating tumor cells (CTCs). Capturing, enumerating and analyzing the genetic and metabolomic characteristics of these CTCs may hold the key for guiding doctors to better understand the source of cancer at an earlier stage for more efficacious disease management.

The isolation of CTCs from whole blood, however, remains a significant challenge due to their (i) low abundance, (ii) lack of a universal surface marker and (iii) epithelial-mesenchymal transition that down-regulates common surface markers (e.g., EpCAM), reducing their likelihood of detection via positive selection assays. These factors potentiate the need for an improved cell isolation strategy that can collect CTCs via both positive and negative selection modalities as to avoid the reliance on a single marker, or set of markers, for more accurate enumeration and diagnosis.

The technologies proposed herein offer a unique set of strategies to focus, sort and template cells in three independent microfluidic modules. The first module exploits ultrasonic standing waves and a class of elastomeric particles for the rapid and discriminate sequestration of cells. This type of cell handling holds promise not only in sorting, but also in the isolation of soluble markers from biofluids. The second module contains components to focus (i.e., arrange) cells via forces from acoustic standing waves and separate cells in a high throughput fashion via free-flow magnetophoresis. The third module uses a printed array of micromagnets to capture magnetically labeled cells into well-defined compartments, enabling on-chip staining and single cell analysis. These technologies can operate in standalone formats, or can be adapted to operate with established analytical technologies, such as flow cytometry. A key advantage of these innovations is their ability to process erythrocyte-lysed blood in a rapid (and thus high throughput) fashion. They can process fluids at a variety of concentrations and flow rates, target cells with various immunophenotypes and sort cells via positive (and potentially negative) selection. These technologies are chip-based, fabricated using standard clean room equipment, towards a disposable clinical tool. With further optimization in design and performance, these technologies might aid in the early detection, and potentially treatment, of cancer and various other physical ailments.

Floral and faunal characteristics and content of photosynthetic pigment in soils of northern Victoria Land, Antarctica

Although soil algae are among the main primary producers in most terrestrial ecosystems of continental Antarctica, there are very few quantitative studies on their relative proportion in the main algal groups and on how their distribution is affected by biotic and abiotic factors. Such knowledge is essential for understanding the functioning of Antarctic terrestrial ecosystems. We therefore analyzed biological soil crusts from northern Victoria Land to determine their pH, electrical conductivity (EC), water content (W), total and organic C (TC and TOC) and total N (TN) contents, and the presence and abundance of photosynthetic pigments. In particular, the latter were tested as proxies for biomass and coarse-resolution community structure. Soil samples were collected from five sites with known soil algal communities and the distribution of pigments was shown to reflect differences in the relative proportions of Chlorophyta, Cyanophyta and Bacillariophyta in these sites. Multivariate and univariate models strongly indicated that almost all soil variables (EC, W, TOC and TN) were important environmental correlates of pigment distribution. However, a significant amount of variation is independent of these soil variables and may be ascribed to local variability such as changes in microclimate at varying spatial and temporal scales. There are at least five possible sources of local variation: pigment preservation, temporal variations in water availability, temporal and spatial interactions among environmental and biological components, the local-scale patchiness of organism distribution, and biotic interactions.

Color reflectance and XRF drill core scanner data of ANDRILL core AND-2A

The compositional record of the AND-2A drillcore is examined using petrological, sedimentological, volcanological and geochemical analysis of clasts, sediments and pore waters. Preliminary investigations of basement clasts (granitoids and metasediments) indicate both local and distal sources corresponding to variable ice-volume and ice-flow directions. Low abundance of sedimentary clasts (e.g., arkose, litharenite) suggests reduced contributions from sedimentary covers while intraclasts (e.g., diamictite, conglomerate) attest to intrabasinal reworking. Volcanic material includes pyroclasts (e.g., pumice, scoria), sediments and lava. Primary and reworked tephra layers occur within the Early Miocene interval (1093 to 640 metres below sea floor mbsf). The compositions of volcanic clasts reveal a diversity of alkaline types derived from the McMurdo Volcanic Group. Finer-grained sediments (e.g., sandstone, siltstone) show increases in biogenic silica and volcanic glass from 230 to 780 mbsf and higher proportions of terrigenous material c. 350 to 750 mbsf and below 970 mbsf. Basement clast assemblages suggest a dominant provenance from the Skelton Glacier - Darwin Glacier area and from the Ferrar Glacier - Koettlitz Glacier area. Provenance of sand grains is consistent with clast sources. Thirteen Geochemical Units are established based on compositional trends derived from continuous XRF scanning. High values of Fe and Ti indicate terrigenous and volcanic sources, whereas high Ca values signify either biogenic or diagenic sources. Highly alkaline and saline pore waters were produced by chemical exchange with glass at moderately elevated temperatures.

The distribution of meiofauna in surface sediments of the Fladen Ground, North Sea

A general study of structure, biomass, and dynamic estimates on meiofauna was carried out during PREFLEX (1975) and FLEX (1976), in 117- 141 m water depth. On the basis of these data an attempt was made to estimate meiofauna production, and this is discussed in relation to the energy input from the spring phytoplankton bloom. Sampling was performed at five stations, but only the stations 1, 4, and 5 were covered by a complete series from August 1975 to July 1976. At each station, from four replicate box core samples, two were withdrawn to study the abundance, distribution, and biomass of meiofauna, the content of chloroplastic pigment equivalents (CPE), and chemical and grain size analyses. At all stations grain size fell in the range of fine sand having median diameters (MD) of < 125 µm. From station 1 to 5 an increase in MD was observed. Highest values of CPE (7.81 µg m l**-1) and organic matter (4.7 %) were obtained in June and July (1976)/ August (1975), respectively. Meiofauna abundance was fairly uniform at all stations examined. Station 1 displayed maximal numbers during the whole investigation period. The abundance per 100 cm**2 varied between 15,550 and 34,900 organisms. All meiofauna studied both in total and as separate taxa showed annual cycles of abundance. Low abundance values were recorded during early summer, and maximum values during winter. High numbers of Foraminifera were obtained for August 1975 (9,460 per 100 cm**2) and July 1976 (9,710 per 100 cm**2). From December to June the values decreased from 3,280 to 1,030 per 100 cm**2. At station 1 maximum values of meiofauna biomass were recorded ranging from 1.5 to 2.7 g DWT m**-2. The mean meiofauna dry weight amounted to 2.1 g DWT m**-2. Based on minimum production, the P/B ratio for the area of station 1 might have a mean of 1.4. Taking into consideration generation times we believe that a turnover ratio of 2 is a conservative value for the Fladen Ground meiofauna. The annual production would amount to 4.2 g DWT m**-2 yr**-1. This is 27.5 % of the energy supply during the spring phytoplankton bloom, which is channelled into the meiofauna. The hypothesis is put forward that the energetic strategy of deep offshore meiofauna differs distinctively from that of shallow inshore meiofauna. While the shallow inshore meiofauna show a relatively fast response to organic matter input, the deep offshore meiofauna reacts much more slowly, the food energy consumption seems to be spread out over a longer period.

The evolution of imperfect mimicry in hoverflies

Ideas about the evolution of imperfect mimicry are reviewed. Their relevance to the colours patterns of hoverflies (Diptera, Syrphidae) are discussed in detail. Most if not all of the hoverflies labelled as mimetic actually are mimics. The apparently poor nature of their resemblance does not prevent them from obtaining at least some protection from suitably experienced birds. Mimicry is a dominant theme of this very large family of Diptera, with at least a quarter of all species in Europe being mimetic. Hoverfly mimics fall into three major groups according to their models, involving bumblebees, honeybees and social wasps. There are striking differences in the general levels of mimetic fidelity and relative abundances of the three groups, with accurate mimicry, low abundance and polymorphism characterizing the bumblebee mimics: more than half of all the species of bumblebee mimics are polymorphic. Mimics of social wasps tend to be poor mimics, have high relative abundance, and polymorphism is completely absent. Bumblebee models fall into a small number of Muellerian mimicry rings which are very different between the Palaearctic and Nearctic regions. Social wasps and associated models form one large Muellerian complex. Together with honeybees, these complexes probably form real clusters of forms as perceived by many birds. All three groups of syrphid mimics contain both good and poor mimics; some mimics are remarkably accurate, and have close morphological and behavioural resemblance. At least some apparently 'poor' mimetic resemblances may be much closer in birds' perception than we imagine, and more work needs to be done on this. Bumblebees are the least noxious and wasps the most noxious of the three main model groups. The basis of noxiousness is different, with bumblebees being classified as non-food, whereas honeybees and wasps are nasty-tasting and (rarely) stinging. The distribution of mimicry is exactly what would be expected from this ordering, with polymorphic and accurate forms being a key feature of mimics of the least noxious models, while highly noxious models have poor-quality mimicry. Even if the high abundance of many syrphid mimics relative to their models is a recent artefact of man-made environmental change, this does not preclude these species from being mimics. It seems unlikely that bird predation actually controls the populations of adult syrphids. Being rare relative to a model may have promoted or accelerated the evolution of perfect mimicry: theoretically this might account for the pattern of rare good mimics and abundant poor ones, but the idea is intrinsically unlikely. Many mimics seem to have hour-to-hour abundances related to those of their models, presumably as a result of behavioural convergence. We need to know much more about the psychology of birds as predators. There are at least four processes that need elucidating: (a) learning about the noxiousness of models; (b) the erasing of that learning through contact with mimics (extinction, or learned forgetting); (c) forgetting; (d) deliberate risk-taking and the physiological states that promote it. Johnston's (2002) model of the stabilization of imperfect mimicry by kin selection is unlikely to account for the colour patterns of hoverflies. Sherratt's (2002) model of the influence of multiple models potentially accounts for all the patterns of hoverfly mimicry, and is the most promising avenue for testing.