Emergent synergistic lysosomal toxicity of chemical mixtures in molluscan blood cells (hemocytes).

The problem of effective assessment of risk posed by complex mixtures of toxic chemicals in the environment is a major challenge for government regulators and industry. The biological effect of the individual contaminants, where these are known, can be measured; but the problem lies in relating toxicity to the multiple constituents of contaminant cocktails. The objective of this study was to test the hypothesis that diverse contaminant mixtures may cause a greater toxicity than the sum of their individual parts, due to synergistic interactions between contaminants with different intracellular targets. Lysosomal membrane stability in hemocytes from marine mussels was used for in vitro toxicity tests; and was coupled with analysis using the isobole method and a linear additive statistical model. The findings from both methods have shown significant emergent synergistic interactions between environmentally relevant chemicals (i.e., polycyclic aromatic hydrocarbons, pesticides, biocides and a surfactant) when exposed to isolated hemocytes as a mixture of 3 & 7 constituents. The results support the complexity-based hypothesis that emergent toxicity occurs with increasing contaminant diversity, and raises questions about the validity of estimating toxicity of contaminant mixtures based on the additive toxicity of single components. Further experimentation is required to investigate the potential for interactive effects in mixtures with more constituents (e.g., 50-100) at more environmentally realistic concentrations in order to test other regions of the model, namely, very low concentrations and high diversity. Estimated toxicant diversity coupled with tests for lysosomal damage may provide a potential tool for determining the toxicity of estuarine sediments, dredge spoil or contaminated soil.

Optimization of foraging and diet by the piscivorous Othos dentex (Serranidae).

The aim of this study was to determine the dietary characteristics and mouth morphology of Othos dentex and to use these data, together with in situ observations of feeding behaviour, to elucidate how foraging and diet are optimized by this piscivorous serranid. Seasonal spear and line fishing over reefs in south-western Australia yielded 426 O. dentex (total length, LT , 183-605 mm), among which the stomachs of 95 contained food. The food in the stomachs of 76 fish was sufficiently undigested to be seen to contain, almost invariably, a single fish prey, which was typically identifiable to family and often to species. The prey of O. dentex, which were measured (LT ), represented 10 families, of which the Labridae and Pempheridae constituted nearly two-thirds of the prey volume. Two-way crossed analysis of similarities of volumetric data for stomach contents showed that the dietary compositions of the different length classes of O. dentex in the various seasons were significantly related to length class of prey, but not to prey family, length class within the various prey families or season. Furthermore, an inverse (Q-mode) analysis, including one-way analysis of similarities, showed that the patterns in the prey consumed by the different length classes of O. dentex in the various seasons were related more strongly to length class than prey family. The former trend is exemplified in a shade plot, by a marked diagonality of the length classes of prey with increasing predator size. The ingestion of typically a single teleost prey, whose body size increases as that of O. dentex increases, reduces the frequency required for seeking prey, thus saving energy and reducing the potential for intraspecific competition for food. The ability of O. dentex to ingest large prey is facilitated by its possession of a very large gape, prominent recurved teeth, dorsal and independently-moveable eyes, cryptic colouration and effective ambush behaviour. Othos dentex has thus evolved very cost-effective mechanisms for optimizing its foraging and diet.

Interdecadal changes in the community, population and individual levels of the fish fauna of an extensively modified estuary.

This study examined inter-period changes over two to three decades in the fish fauna of an urbanized estuary experiencing rapid population growth and a drying climate (Swan-Canning Estuary, Western Australia). Responses were compared at the fish community level (species composition; 1978-2009 in the shallows and 1993-2009 in deeper waters) and at the population and individual levels of an estuarine indicator species, black bream Acanthopagrus butcheri (biomass-abundance and per capita mass at age, respectively; 1993-2009). All three levels showed distinct shifts from earlier to later periods, but their patterns, sensitivity and breadth differed. Community composition changed markedly in the shallows of the lower-middle estuary between the late 1970s and all later periods and moderately between more disparate periods from 1995 to 2009. Several species trends could be linked to the increasing salinity of the estuary or declining dissolved oxygen levels in its middle-upper reaches. Community changes were, however, small or insignificant in the shallow and deeper waters of the upper estuary and deeper waters of the middle estuary, where environmental perturbations are often most pronounced. This may reflect the resilience of the limited suite of species that typify those reaches and thus their lack of sensitivity in reflecting longer-term change at the coarser level of mean abundance. One such species, the selected indicator, A. butcheri, did, however, show marked temporal changes at both the population and individual levels. Biomass decreased markedly in deeper waters while increasing in the shallows from earlier to later periods, presumably reflecting an onshore movement of fish, and per capita body mass in the 2+, 3+ and 4+ year classes fell steadily over time. Such changes probably indicate deteriorating habitat quality in the deeper waters. The study outcomes provide support for a multifaceted approach to the biomonitoring of estuaries using fishes and highlight the need for complementary monitoring of relevant stressors to better disentangle cause-effect pathways.

Dietary variations within a family of ambush predators (Platycephalidae) occupying different habitats and environments in the same geographical region.

This study has determined the extents and basis for variations in the composition of the prey ingested by the abundant species of a family highly adapted for ambush predation, i.e. Platycephalidae, in a region (south-western Australia) where that family is found in different habitats and environments. Dietary data were thus collected for Leviprora inops and Platycephalus laevigatus from seagrass in marine embayments and for Platycephalus westraliae from over sand in an estuary. These were then collated with those recorded previously for Platycephalus speculator from over sand and in seagrass in an estuary and for Platycephalus longispinis from over sand in coastal marine waters. While crustaceans and teleosts together dominated the diet of all five species, their percentage volumetric dietary contributions varied greatly, with those of crustaceans ranging from 7% for L. inops to 65% for P. speculator and those of teleosts ranging from 29% for P. longispinis to 91% for L. inops. For analyses, the data were separated into two sets. The first comprised the 17 dietary categories of invertebrates and all identified and unidentified teleosts collectively, while the second consisted of the 23 identified teleost families, both of which were subjected to permutational analysis of variance (PERMANOVA), analysis of similarities (ANOSIM) and a new (two-way) version of the RELATE procedure. The diets of three species changed seasonally, when using invertebrate dietary categories and teleosts collectively, but with only one species, when employing identified teleost families, probably reflecting a greater tendency for invertebrate than teleost prey abundance to change during the year. On the basis of dietary data for invertebrate taxa + teleosts collectively, the diets of three of the five species changed serially with body size, with a fourth species feeding, throughout life, predominantly on the carid Palaemonetes australis. Based on identified teleost families, the diets of the three species that fed predominantly on teleosts underwent serial size-related changes. Although L. inops and the co-occurring P. laevigatus both consume large volumes of teleosts, the former ingests larger, less demersal and more mobile prey, e.g. the labrids Haletta semifasciata and Neoodax balteatus, than the latter, e.g. the scorpaenid Gymnapistes marmoratus, reflecting the possession by L. inops of a far longer head and larger buccal cavity. Circumstantial evidence suggests that the large differences in the volumes of crustaceans and teleosts consumed by each platycephalid species are related to differences in the relative availability of these prey in the different habitats or environments of each species.

Characteristics of the ichthyofauna of a temperate microtidal estuary with a reverse salinity gradient, including inter-decadal comparisons.

Data on the fish fauna of the Leschenault Estuary on the lower west coast of Australia were collected and used as a model to elucidate the characteristics of permanently open estuaries with a reverse salinity gradient, which undergo seasonal changes similar to many other estuaries with Mediterranean climate. Focus was placed on determining (1) the relationships of the number of species, density, life cycle category and species composition of fishes with region (within estuary), season and year and salinity, (2) whether species are partitioned along the lengths of such systems and (3) the extent and significance of any inter-decadal changes in species composition. The analyses and interpretation involved using multi-factorial permutational multivariate analysis of variance (PERMANOVA) and analysis of similarity (ANOSIM) designs, and three new or recently published visualization tools, i.e. modified non-metric multidimensional scaling (nMDS) plots, coherent species curves and segmented bubble plots. The base, lower, upper and apex regions of the Leschenault Estuary, along which the salinity increased in each season except in winter when most rainfall occurs, were sampled seasonally for the 2 years between winter 2008 and autumn 2010. Estuarine residents contributed twice as many individuals, but less than half the number of species as marine taxa. While the numbers of marine species and estuarine residents declined between the base or lower and apex regions, the individuals of marine species dominated the catches in the base region and estuarine residents in the other three regions. Ichthyofaunal composition in each region underwent conspicuous annual cyclical changes, due to time-staggered differences in recruitment among species, and changed sequentially along the estuary, both paralleling salinity trends. Different groups of species characterized the fauna in the different regions and seasons, thereby partitioning resources among species. The ichthyofauna of the apex region, in which salinities reached 54 and temperatures 36° C, recorded the highest maximum density and, in terms of abundance, was dominated (90%) by three atherinid species, emphasizing the ability of this family to tolerate extreme conditions. Comparisons between the data for 2008-2010 and 1994 demonstrate that the spotted hardyhead Craterocephalus mugiloides and the common hardyhead Atherinomorus vaigiensis had colonized and become abundant in the Leschenault Estuary in the intervening period. This represents a southwards extension of the distribution of these essentially tropical species during a period of increasing coastal water temperatures as a result of climate change. The abundance of weed-associated species, e.g. the western gobbleguts Ostorhinchus rueppellii and the soldier Gymnapistes marmoratus, increased, whereas that of the longfinned goby Favonigobius lateralis decreased, probably reflecting increases in eutrophication and siltation, respectively.

To what extent are the dietary compositions of three abundant, co-occurring labrid species different and related to latitude, habitat, body size and season?

This study demonstrated that the dietary composition of each of three abundant reef-associated labrid species in temperate Western Australia differed significantly with latitude and changed with increasing body size and almost invariably differed among those species when they co-occurred. These results were derived from comparisons and multivariate analyses of volumetric dietary data, obtained from the foregut contents of Coris auricularis, Notolabrus parilus and Ophthalmolepis lineolatus from the Jurien Bay Marine Park (JBMP) and waters off Perth, 250 km to the south. Latitudinal differences in the dietary compositions of each species in exposed reefs typically reflected greater contributions by large crustaceans, bivalve molluscs, echinoids and annelids to the diets in the waters off Perth than in the JBMP, whereas the reverse was true for gastropods and small crustaceans. The diet of each species exhibited similar, but not identical, quantitative changes with increasing body size, with the contributions of small crustaceans declining and those of large crustaceans and echinoids increasing, while that of gastropods underwent little change. Within the JBMP, the dietary compositions of both C. auricularis and N. parilus were similar in exposed and sheltered reefs and the same was true for N. parilus in the sheltered reefs and interspersed areas of seagrass. The latter similarity demonstrated that, in both of those divergent habitat types, N. parilus feeds on prey associated with either the sand or the macrophytes that cover and lie between the reefs. Although the main dietary components of each species were the same, i.e. gastropods, small crustaceans (mainly amphipods and isopods), large crustaceans (particularly penaeids and brachyuran crabs) and echinoids, their contributions varied among those species, which accounts for the significant interspecific differences in diet. Coris auricularis had the most distinct diet, due mainly to an ingestion of greater volumes of small crustaceans, e.g. amphipods and isopods, and lesser volumes of large crustaceans, e.g. brachyuran crabs, which was associated with a relatively narrower mouth and smaller teeth and the absence of prominent canines at the rear of the jaw. The above intra and interspecific differences in dietary composition would reduce, on the south-west coast of Australia, the potential for competition for food among and within these three abundant labrids, each of which belongs to different genera within the Julidine clade.

Benthic macroinvertebrates as indicators of environmental deterioration in a large microtidal estuary.

This study tested the hypothesis that, during recent years, the benthic macroinvertebrate fauna of the large basin of the Swan-Canning Estuary has changed in ways consistent with deteriorating environmental conditions in that estuary. Between 1986/7 and 2003/4, the compositions of that fauna altered markedly at the species and even family levels. Thus, the densities and number of species of molluscs, and especially of crustaceans, which are particularly susceptible to environmental stress, declined, while those of the more tolerant polychaetes increased. However, taxonomic distinctness declined consistently only at one of the four widely-spaced sampling sites and the dispersion of samples did not differ markedly between periods, indicating that the benthic fauna has not undergone such extreme changes as in the nearby Peel-Harvey Estuary. It is thus proposed that benthic macroinvertebrates can act as important indicators of the severity of environmental degradation in microtidal estuaries in regions where such perturbations are increasing.

Changes in the benthic macroinvertebrate fauna of a large microtidal estuary following extreme modifications aimed at reducing eutrophication.

An artificial channel was opened in 1994 between the microtidal Peel-Harvey Estuary and the Indian Ocean to increase tidal exchange and thus ameliorate the problems of eutrophication. Although this greatly reduced macroalgal and cyanobacterial growths and the amount of particulate organic matter, our data indicate that, contrary to managerial expectations, the benthic environment has deteriorated. Thus, although macroinvertebrate density has declined as predicted, taxonomic distinctness (Delta( *)) has also declined and species composition has become more variable. Macroinvertebrate composition has also changed markedly at the species, family and even phylum levels. The Crustacea, the most sensitive of the major macrobenthic taxa to environmental stress, has become proportionally less abundant and speciose, whereas the Polychaeta, the least sensitive, was unique in showing the reverse trend. The benthos of the Peel-Harvey Estuary is thus apparently more stressed than previously, probably due to the multiple effects of a great increase in system use.

Acute anhydrous ammonia intoxication in cattle.

Intoxication with anhydrous ammonia (AA) is a common occupation-related health problem affecting farmers and fertilizer applicators, however, very few descriptions of animal exposure to this toxicant exist. Thieves entered a feedlot and damaged a liquid AA storage tank, resulting in the accidental leakage of gas from a valve. Overnight, 12 cattle were found dead, with a total mortality of 64 cattle out of 260 on the premises dying or euthanized in < 1 week. Signs of affected cattle included blindness, drooling, inappetence, respiratory distress, recumbency, and death. Two cattle were submitted live for diagnostic evaluation 3 days after initial exposure. Gross lesions included corneal ulcers, fibrinonecrotizing rhinitis, hemorrhages within the nasal sinuses, and anterioventral bronchopneumonia. Microscopic lesions consisted predominantly of degeneration and necrosis of superficial epithelium lining nasal passages, trachea, and pulmonary airways. This case illustrates the acute AA effects on cattle, which is likely to be an increasingly encountered problem because of on-farm storage of AA and its access by illicit drug manufacturers.

Experimental inoculation of wild turkeys (Meleagris gallopavo) with Mycobacterium bovis.

Although avian species are known to be susceptible to infection with Mycobacterium spp. organisms, much remains unknown about the susceptibility of birds to infection with M. bovis. The objective of this current study was to determine if wild turkeys (Meleagris gallopavo) can be infected with M. bovis when inoculated by the oral or intratracheal route. Six turkeys were orally inoculated and another six were inoculated via the trachea with a high dose of M. bovis, 1 x 10(5) CFU/ml. Six turkeys were sham-inoculated controls. Two turkeys from each treatment group were sacrificed on days 30, 60, and 90 postinoculation. There were no gross or microscopic lesions consistent with mycobacteriosis in the 23 inoculated turkeys over the 90-day duration of this study. Fecal cultures were also consistently negative for M. bovis when sampled before inoculation and on days 1, 30, and 60 postinoculation. Two intratracheally inoculated turkeys were positive for M. bovis in visceral tissues at 30 days postinoculation. However, this finding was only indicative of passive persistence of mycobacteria in the tissues and not of infection, as there were no attendant lesions or clinical compromise to support infection. Thus, it can be concluded that young wild turkeys are resistant to infection with M. bovis and, therefore, pose minimal threat as reservoir or spillover hosts for this organism.

Ecological consequences of dredged material disposal in the marine environment: a holistic assessment of activities around the England and Wales coastline.

This study provides a holistic perspective on the ecological effects of dredged material disposal, both intertidally and subtidally. A number of numerical techniques (univariate, distributional, multivariate and meta-analysis) were used to assess impacts at 18 different disposal sites. The analyses revealed that ecological effects associated with dredged material disposal were dependent on the numerical techniques used, and that impacts were disposal-site specific. Disposal-site communities were generally faunistically impoverished to varying degrees, and impacts following intertidal placement were comparable to those of subtidal placement. We conclude that any assessment of the consequences of dredged material disposal to the coastal environment must take account of site-specific variation in prevailing hydrographic regimes and in ecological status, along with information on the disposal activity itself (mode, timing, quantity, frequency and type of material). As would be expected, variability in the latter presents a significant challenge in attempts to generalise about environmental and ecological impacts.

Resistance of Mallard ducks (Anas platyrhynchos) to experimental inoculation with Mycobacterium bovis.

The purpose of this study was to investigate whether mallard ducks (Anas platyrhynchos) are susceptible to infection with Mycobacterium bovis by either oral or intratracheal inoculation and to assess their potential role in the spread of bovine tuberculosis. Six ducks were orally inoculated with 1.0 x 10(5) colony-forming units of M. bovis, six ducks were intratracheally inoculated with the same dose, and six ducks served as sham-inoculated controls. The study length was 90 days postinoculation, with samples of two birds from each group necropsied at 30-day intervals. Both fecal and tissue samples were collected for mycobacterial culture. None of the inoculated ducks shed M. bovis in their feces at any culture point (days 1, 30, and 60) during the study. No evidence of illness or weight loss was present during the course of the study, and only one duck had M. bovis isolated from any tissue, although there were no associated microscopic lesions. Mallard ducks were highly resistant to infection with M. bovis following high-dose inoculation and did not shed the organism in their feces. This study was conducted using high-dose inoculation; therefore, it appears that ducks are unlikely to play any significant role in the transmission of M. bovis between infected and uninfected mammalian hosts.

Plant-like mating in an animal: sexual compatibility and allocation trade-offs in a simultaneous hermaphrodite with remote transfer of sperm.

The importance of sexual compatibility between mates has only recently been realized in zoological research into sexual selection, yet its study has been central to botanical research for many decades. The reproductive characteristics of remote mating, an absence of precopulatory mate screening, internal fertilization and embryonic brooding are shared between passively pollinated plants and a phylogenetically diverse group of sessile aquatic invertebrates. Here, we further characterize the sexual compatibility system of one such invertebrate, the colonial ascidian Diplosoma listerianum. All 66 reciprocal pairings of 12 genetic individuals were carried out. Fecundities of crosses varied widely and suggested a continuous scale of sexual compatibility. Of the 11 animals from the same population c. 40% of crosses were completely incompatible with a further c. 20% having obvious partial compatibility (reduced fecundity). We are unaware of other studies documenting such high levels of sexual incompatibility in unrelated individuals. RAPD fingerprinting was used to estimate relatedness among the 12 individuals after a known pedigree was successfully reconstructed to validate the technique. In contrast to previous results, no correlation between genetic similarity and sexual compatibility was detected. The blocking of many genotypes of sperm is expected to severely modify realized paternity away from 'fair raffle' expectations and probably reduce levels of intra-brood genetic diversity in this obligatorily promiscuous mating system. One adaptive benefit may be to reduce the bombardment of the female reproductive system by outcrossed sperm with conflicting evolutionary interests, so as to maintain female control of somatic : gametic investment.

Tuberculous lesions in free-ranging white-tailed deer in Michigan.

Descriptions of the anatomical distribution of Mycobacterium bovis gross lesions in large samples of white-tailed deer (Odocoileus virginianus) are lacking in the scientific literature. This report describes the distribution of gross lesions in the 58 white-tailed deer that cultured positive for M. bovis among the 19,500 submitted for tuberculosis testing in Michigan (USA) in 1999. For the vast majority (19,348) of those tested, only the head was submitted; for others, only extracranial tissues (33) or both the head and extracranial tissues (119) were available. Among those deer that cultured positive, cranial gross lesions were noted most frequently in the medial retropharyngeal lymph nodes, although solitary, unilateral parotid lymph node lesions also were found. Extracranial lesions occurred most commonly in the thorax. The distribution of lesions largely agreed with the few existing case reports of the M. bovis in white-tailed deer, although gross lesions were also found in sites apparently not previously reported in this species (liver, spleen, rumen, mammary gland). Some practical issues that may assist future surveillance and public education efforts are also discussed.

Hepatic metallothionein as a biomaker for metal contamination: age effects and seasonal variation in European flounders (Pleuronectes flesus) from the Severn Estuary and Bristol Channel.

Hepatic concentrations of metallothionein [MT] and three metals (Cu, Zn, Cd) were determined in 242 European flounders (Pleuronectes flesus) collected from power stations at Oldbury-upon-Severn and Hinkley Point, located in Severn Estuary and Bristol Channel, UK, respectively, between March 1996 and February 1998. A model involving three-factor analysis of variance (ANOVA) was used to examine variation in MT and metal concentrations with respect to season, year and site; with age-class included as a covariate in the analysis. Hepatic concentrations of MT and Cd (and to some degree, Cu, but not Zn) increased significantly with age. The model explained 38, 25, 17 and 26% of the variation in MT, Cu, Zn and Cd, respectively, with significant effects due to season, and to a lesser extent, to year. Site was only a significant factor for Cd which was higher in fish from Hinkley. Correlation between the individual concentration of MT and each metal alone, or in combination, was poor, and explained only an additional 3.0% of the residual variation in MT, most of which was attributable to Cu (2.7%). Compared to other industrialised estuaries, Cd concentrations were high (>20 micro g-1 in some individuals). The study emphasises the importance of seasonal variation and other factors in biomonitoring programmes and highlights the limitations of using [MT] as a biomarker for metal contamination in flounders from the Severn Estuary.

Fish fauna of the Severn Estuary. Are there long-term changes in abundance and species composition and are the recruitment patterns of the main marine species correlated?

Fish were collected from the intake screens of the Oldbury Power Station in the Severn Estuary in each week between early July 1972 and late June 1977 and at least twice monthly between early January 1996 and late June 1999. The annual catches, after adjustment to a common sampling effort, demonstrate that the abundance of fish at Oldbury was far greater in the 1990s than 1970s, mainly due to marked increases in the numbers of certain marine species, such as sand goby, whiting, bass, thin-lipped grey mullet, herring, sprat and Norway pout. These increases may reflect the great improvement that occurred in the water quality of the Severn Estuary between these decades. The only species that declined markedly in abundance was poor cod. Modest declines in flounder and River lamprey paralleled those occurring elsewhere in the UK. The species composition in the two decades also differed, reflecting changes not only in the relative abundances of the various marine estuarine-opportunistic species, which dominated the ichthyofauna, but also in those of the suite of less abundant species in the estuary. The cyclical changes undergone each year by the species composition of the fish fauna of the Severn Estuary reflect sequential intra-annual changes in the relative abundances of species representing each of the marine, diadromous and freshwater categories. New approaches have been developed to test whether or not large sets of correlations between patterns of recruitment amongst abundant marine species (internal correlations), and between those patterns and salinity and water temperature within the estuary (cross-correlations), were significant. The correlation profile analyses found no evidence that the annual recruitment strengths of these species were either intercorrelated, or correlated with either one or a combination of both of the above environmental variables. Yet, the timings of the recruitment of these species into the estuary were intercorrelated, i.e. a slightly earlier or later than normal immigration by one species in a given year was paralleled by the same trend in other species. However, this association in recruitment times could be linked neither to salinity nor water temperature within the estuary, nor to a combination of these two variables. These results indicate that, while the factors that influence the annual recruitment strengths of the juveniles of different marine species vary, inter-annual differences in the phasing of events that regulate spawning times and/or larval dispersal influence, in the same direction, the times when marine species are recruited into the estuary.

Comparison of postmortem techniques for the detection of Mycobacterium bovis in white-tailed deer (Odocoileus virginianus).

A retrospective study of various diagnostic postmortem techniques used in a 4-year surveillance program for detection of Mycobacterium bovis infection in wild white-tailed deer (Odocoileus virginianus) was conducted. The tests evaluated were routine histopathology, acid-fast staining, detection of acid-fast bacilli in culture, and an M. tuberculosis group-specific genetic probe applied to pure cultures. Each of these techniques were compared with a reference or "gold standard" of mycobacterial culture and identification. Histopathology, the most rapid form of testing for M. bovis infection in white-tailed deer samples, had a sensitivity of 98% and a specificity of 87%, resulting in a positive predictive value of 94%. The detection of acid-fast bacilli by staining was less sensitive than histopathology (90%), but its higher specificity (97%) resulted in a positive predictive value of 99%. The detection of acid-fast bacilli on culture was both highly specific (93%) and sensitive (100%). The group-specific genetic probe had the highest sensitivity and specificity and produced results in complete agreement with those of mycobacterial culture, suggesting that this technique could be used as the new "gold standard" for this particular wildlife tuberculosis surveillance program.

Quantifying structural redundancy in ecological communities.

In multivariate analyses of the effects of both natural and anthropogenic environmental variability on community composition, many species are interchangeable in the way that they characterise the samples, giving rise to the concept of structural redundancy in community composition. Here, we develop a method of quantifying the extent of this redundancy by extracting a series of subsets of species, the multivariate response pattern of each of which closely matches that for the whole community. Structural redundancy is then reflected in the number of such subsets, which we term "response units", that can be extracted without replacement. We have applied this technique to the effects of the Amoco-Cadiz oil-spill on marine macrobenthos in the Bay of Morlaix, France, and to the natural interannual variability of macrobenthos at two stations off the coast of Northumberland, England. Structural redundancy is shown to be remarkably high, with the number and sizes of subsets being comparable in all three examples. Taxonomic/functional groupings of species within the differing response units change in abundance in the same way over time. The response units are shown to possess a wide taxonomic spread and, using two different types of randomisation test, demonstrated to have a taxonomically and functionally coherent structure. The level of structural redundancy may therefore be an indirect measure of the resilience or compensation potential within an assemblage.

Methodology for estimating numbers of free-living and attached bacteria in estuarine water.

A fundamental problem in estuarine microbiology studies is the accurate determination of the density in the water column of both free-living bacteria and those attached to suspended particulate matter. When a water sample is filtered and the filter is viewed by epifluorescence microscopy, counts can be made of the numbers of bacteria which are seen on the filter background (free-living) and those which appear to lie on sediment particles (both free-living and attached). With only the additional knowledge of the proportion of the filter area covered by particles (a quantity that is straightforwardly determined by stereological point counting), results from geometric probability were used to determine the expected number of bacteria which are hidden by particles and hence to provide an estimation scheme for the true densities of free-living and attached bacteria. Variance equations based on a Taylor series are given, and a partial check of the method is attempted with controlled mixtures of bacteria and sediment. An alternative procedure is also proposed, in which the natural attached/free-living ratio is altered by an intervention experiment, allowing an estimation which is less model dependent but more labor intensive. Both methods are applied to a series of samples from the Tamar estuary, United Kingdom, taken in April 1985. A notable conclusion is that there are always more free-living than attached bacteria in the water column throughout the estuary.

An investigation of adhesion and detachment in slime mould amoebae using columns of hydrophobic beads.

Vegetative amoebae of Dictyostelium discoideum (strain AX2) adhere to columns of hydrophobic beads (Bio-Beads SM-2) which have been coated with axenic growth medium. This binding is blocked by sodium azide and other inhibitors of aerobic respiration. Bound cells are not detached by lowering the temperature but they can be eluted from the column by including azide in the medium. However, this effect is critically influenced by temperature. The elution profiles become progressively shallower as the temperature is lowered. The profiles represent a set of probability distributions whose means and variances increase with decreasing temperature. Below 20 degrees C, the mean time to cell detachment is a linear function of temperature. A statistical model of this azide-stimulated detachment has been developed which closely predicts the real data. It assumes that each amoeba has k binding sites whose stochastically independent behaviour is governed by a parameter theta; the mean time to detachment of each site is 1/theta. Both parameters have been estimated. The value of k is close to 3 and does not vary with temperature below 20 degrees C. In contrast, theta varies in such a way that the value of 1/theta increases linearly with decreasing temperature, so paralleling the behaviour of the mean cell detachment time.