Identification of a Microsporidian Polar Tube Protein Reactive Monoclonal Antibody

ABSTRACT. The microsporidia are characterized by spores containing a single polar tube that coils around the sporoplasm. When triggered by appropriate stimuli, the polar tube rapidly discharges out of the spore forming a hollow tube. The sporoplasm passes out of the spore through this tube serving as a unique vehicle of infection. Due to the unusual functional and solubility properties of the polar tube, the proteins comprising it are likely to be members of a protein family with a highly conserved amino acid composition among the various microsporidia. Polar tube proteins were separated from the majority of other proteins in glass bead disrupted spores of Glugea americanus using sequential 1% sodium dodecyl sulfate (SDS) and 9M urea extractions. The resultant spore pellet demonstrated broken, empty spore coats and numerous polar tubes in straight and twisted formations by negative stain transmission electron microscopy. After subsequent incubation of the pellet with 2% dithiothreitol (DTT), empty spore coats were still observed but the polar tubes were no longer present in the pellet. The DTT supernatant demonstrated four major protein bands by SDS-PAGE: 23, 27, 34 and 43 kDa. Monoclonal antibodies were produced to these proteins using Hunter's Titermax adjuvant. Mab 3C8.23.1 which cross-reacted with a 43-kDa antigen by immunoblot analyis, demonstrated strong reactivity with the polar tube of G. americanus spores by immunogold electron microscopy. This antibody will be useful in further characterization of polar tube proteins and may lead to novel diagnostic and therapeutic reagents.     

Coral reef bleaching: facts, hypotheses and implications

Coral reef bleaching, the temporary or permanent loss of photosynthetic microalgae (zooxanthellae) and/or their pigments by a variety of reef taxa, is a stress response usually associated with anthropogenic and natural disturbances. Degrees of bleaching, within and among coral colonies and across reef communities, are highly variable and difficult to quantify, thus complicating comparisons of different bleaching events. Small-scale bleaching events can often be correlated with specific disturbances (e.g. extreme low/high temperatures, low/high solar irradiance, subaerial exposure, sedimentation, freshwater dilution, contaminants, and diseases), whereas large scale (mass) bleaching occurs over 100s to 1000s of km², which is more difficult to explain. Debilitating effects of bleaching include reduced/no skeletal growth and reproductive activity, and a lowered capacity to shed sediments, resist invasion of competing species and diseases. Severe and prolonged bleaching can cause partial to total colony death, resulting in diminished reef growth, the transformation of reef-building communities to alternate, non-reef building community types, bioerosion and ultimately the disappearance of reef structures. Present evidence suggests that the leading factors responsible for large-scale coral reef bleaching are elevated sea temperatures and high solar irradiance (especially ultraviolet wavelengths), which may frequently act jointly.     

A feasible Global Carbon Cycle Observing System: a plan to decipher today's carbon cycle based on observations

A design is presented to continuously monitor the transfers of carbon between the atmosphere and the terrestrial biosphere and oceans on large spatial scales. A consideration of the expected signal-to-noise suggests that a very cost effective method is to perform repeated measurements of the mixing ratios of trace gases in the atmospheric column at an increased number of sites compared to the present geographical coverage. This can be accomplished with existing technology, provided that more automation is applied to the sampling process as well as to the analytical procedures. The resulting data will be crucial to test extrapolations based on direct flux measurements to regional and global scales.     

Plant coexistence in coastal heaths: Habitat segregation in the post-fire environment

Abstract Previous studies of heaths on Pleistocene coastal sands showed consistent variation in patterns of floristic composition between areas on ridges and slopes but not within them. A large wild-fire that swept this system provided an opportunity to observe temporal processes in habitat segregation of species. Ridges and slopes were found to differ in species richness of seedlings in 0.0625 m2 quadrats and there was no evidence that the species richness in the two habitats was converging over 3 years. This suggests that initially these processes differ in space between the two habitats with more species in the wet heath being packed into a smaller area than in the dry heath. Seed-banks of species did not saturate available space for recruitment in either habitat, but seedling densities differed asymmetrically between habitats across two pairs of species studied. Experimental manipulation of seeds among habitats also showed distinct differences in establishment and survival among representative species from different habitats. The presence of seed-banks of wet-heath species in dry heath suggested that recruitment of their seedlings may occur there, but experiments showed that seedlings that arise from them do not survive there. Conversely, although no seed-bank of either dry-heath species studied was found in wet heath, our manipulations showed that their seedlings could grow there. Thus, superficially, physiological tolerance appeared to limit regeneration of wet-heath species in dry heath. However, there appeared to be no physiological limit for dry-heath plants to survive in wet heath. This paper shows that segregation of species may operate at either of two stages of the life cycle: at dispersal when safe sites for establishment are required, and at recruitment after seedlings have established. These findings highlight the importance of the regeneration niche in structuring community composition in coastal heaths, and contrast with traditional explanations of species segregation invoking physiological tolerance and competition at later life-history stages.     

Risk-Sensitive Foraging, Fitness, and Life Histories: Where Does Reproduction Fit into the Big Picture?

Risk-sensitive foraging may occur whenever feeding success hasnon-linear effects on fitness. Models of sensitivity to variationin amount of food obtained have concentrated on foraging inorder to survive even though feeding during growth, migration,or reproduction can have strong and non-linear effects on fitness.I illustrate how risk-sensitive foraging for reproduction coulddiffer from risk-sensitive foraging for survival using two simplemodels. Each model assumes that organisms must accumulate somethreshold amount of resources before they can reproduce. Inthe first model, additional resources above the threshold leadto increased reproductive success. Here variance in feedingsuccess can be advantageous even when the mean gain would alloworganisms to reproduce. In the second model, early breedingis superior to late breeding because recruitment rate declinesover time. Here a symmetrical distribution in foraging ratesresults in a skewed distribution of breeding times. Despitethis, variance in feeding success may be advantageous even whenthe seasonal decline in recruitment is linear. The advantageof risk is much larger if foragers can switch freely betweenfeeding options. These two models are united by "the bankruptcyeffect" because all failures to reproduce carry the same consequences.These simple models suggest that risk-prone foraging may belikely when factors other than survival are important for fitness.Finally, I advocate using a mix of modelling techniques, notethe implicitly risk-sensitive nature of stochastic dynamic programming,and speculate on potential connections between risk-sensitiveforaging and theories of life histories in variable environments.INTRODUCTION visit restaurants that provided twice     

Disturbance and the community structure of stream invertebrates: patch-specific effects and the role of refugia

1. We have previously shown that the impact of spates on stream invertebrates may differ among patches separated by distances of metres or less. Here we analyse the species-specific flood responses of larval chironomids and adult and near mature copepods living in different patch types. Four patch types (with eight replicates of each) were compared: the sandy mid-channel, fine sediments around dams, coarse sediments around dams, and dam debris. Additionally, since some fine sediment patches had been shown previously to act as flow refugia while others did not, we also examined species-specific responses in refugium vs. non-refugium fine sediment patches. Detrended correspondence analysis was used to test for changes in assemblage structure (species composition and relative abundance). 2. Species richness was not altered in a predictable manner by floods; the least stable patch types (mid-channel and coarse patches) did not necessarily show reduced species richness during the spate. 3. As indicated by the spread of DCA ordination scores, there was generally a high degree of overlap in the species composition among the four patch types. Nevertheless, copepod species composition and relative abundance were more similar among patch types during the spate than pre-spate. Spates may induce a re-distribution of copepod species among the patch types. Chironomid species composition and relative abundance were no more similar among patch types during the spate than pre- or post-spate. 4. For both chironomids and copepods, species composition and relative abundance (as assessed by DCA ordination scores) in refugium patches changed more in response to the spate than in the non-refugium patches. An influx of individuals from just a few species for each group was responsible for the change in assemblage structure. Thus, despite the fact that our past work has shown that refugia may confer enhanced resistance and resilience of copepod and chironomid assemblages in terms of total faunal abundances, the present work suggests that resistance and resilience of the species composition of the community apparently are no greater in refugium patches than in non-refugium patches.     

Fossil record of cladoceran and algal responses to fishery management practices

1. We hypothesized that the fishery management practices of toxaphene application and trout stocking would affect non-target organisms in lakes. Because these practices were rarely monitored in the past, cladoceran and algal assemblages were quantified in sediment cores from two lakes treated 30+ years ago to determine the long-term response of organisms near the base of the food chain. 2. Chydorids were remarkably resistant over the short term (a few years) in both the oligotrophic and eutrophic lakes despite toxaphene treatments that extirpated native fish and other invertebrates. In the oligotrophic lake (Annette Lake), six chydorid taxa were less abundant in the years following treatment, although no loss of species richness was detected. In the eutrophic lake (Chatwin Lake), the dominant Chydorus cf. sphaericus declined coincident with toxaphene treatment, but longer-term declines of all taxa were probably related to food web or other changes rather than to toxaphene toxicity. Cause and effect coupling was complicated by the fact that many chydorids were present at low concentrations in some pretreatment samples. 3. The algal communities (as fossil pigments) responded to treatment differently in the two lakes. In the oligotrophic lake, planktonic diatoms, dinoflagellates and chlorophytes were replaced as dominants by deep-water or benthic blooming cryptophytes, chrysophytes and cyanobacteria. This shift occurred along with increases in large daphnids and the ‘grazing indicator’, pheophorbide a. While both lakes appear to have had enhanced pigment preservation following treatment, the eutrophic lake encountered few long-term changes in its fossil pigment assemblage. Redundancy analysis estimated that the presence or absence of stocked trout explained much of the variation in the algal assemblages, particularly in the oligotrophic lake. 4. Toxaphene remained elevated in profundal sediments from these lakes 30 and 35 years after treatment.     

The anatomy and systematics of the New Caledonian land snail genus Draparnaudia Montrouzier, 1859 (Pulmonata: Orthurethra)

A comprehensive study of the anatomy, ecology and distribution of the orthurethran genus Draparnaudia confirms its family-group status, and suggests a probable sister-group relationship with another Pacific family, the Partulidae; its previous position as a subfamily of the Enidae is refuted. Draparnaudia is endemic to New Caledonia, but records from the New Hebrides are thought to result from introductions. A total of six species are recognized in the taxonomic revision, including two new species. Spermatophores are recorded for the first time.     

Bioassay testing for Australia as part of water quality assessment programmes

Abstract Bioassay testing includes both toxicity (one or more effects is measured) and bioaccumulation (the phenomenon of tissue accumulation is measured) tests. Both types of bioassay tests have an important place in water quality assessment programmes, ideally beginning with initial screening and as part of tiered testing that includes contaminant analyses and field studies. Both have advantages (e. g. measure bioavailability, provide quantitative data, experimental manipulations can address cause-effect relationships) and disadvantages (e. g. laboratory exposures do not necessarily reflect field conditions, not all organisms can be tested). Both provide essential, but not all, information necessary for holistic water quality assessments. This paper provides a review of the use of bioassays including definitions, applications, advantages and disadvantages, utility and relevance. The utility of bioassays in monitoring programmes is illustrated by two case studies. The first case study, involving novel toxicity studies conducted in the high Arctic, illustrates the versatility of bioassays such that testing conducted in a tent on the ice resulted in new knowledge and a change in industrial discharge limits. The second case study further illustrates the importance of bioassays in decision making. In this case, a decision not to immediately spend hundreds of millions of dollars for sewage treatment, but rather to concentrate on source control, was made based in large part on bioassay testing conducted as part of an integrated environmental assessment.     

Can the 15N Dilution Technique be used to Study N2 Fixation in Tropical Tree Symbioses as Affected by Water Deficit?

Three methods were used to study N₂ fixation and effects ofwater deficit on N₂ fixation: C₂H₂ reduction assay (ARA), ¹⁵Ndilution technique and accumulated N content. In addition, ¹⁵Ndilution was calculated both in a traditional way and in a modifiedway, which takes into consideration N and ¹⁵N content for theplants before the experiment started. The three methods wereapplied on the following Rhizobium-symbioses: Acacia albidaDel (Faidherbia albida (Del) A. Chev.) and Leucaena leucocephala(Lam) de Wit., and the Frankia-symbiosis Casuarina equisetifoliaL. The plants wereabout 4-months-old when they were harvested. Nitrogen derived from N₂ fixation in control plants of Acaciaalbida was 54·2 mg as measured with ARA, while it was28·5 mg as measured with the ¹⁵N dilution technique,compared to 30·7 mg calculated as accumulated N. In comparison,L. leucocephala fixed 41·6 mg N (ARA), 53·5 mgN(15N dilution technique) and 56·3 mg N (accumulatedN). The Frankia-symbiosis had fixed 27·4 mg N as measuredby ARA, 8·1 mg N as measured by ¹⁵N dilution techniqueand 12·3 mg N as accumulated N. There were no differencesbetween the estimates based ontraditional and modified waysof calculating ¹⁵N dilution. The immediate effect of water deficit treatment on N₂ fixationwas continuously measured inall species with ARA, which startedto decrease approximately 10 d after the initiation of the treatment,and declined to less than 5% of the initial level after 21–28d. The decrease in the amount of N derived from N₂ fixation wasstudied in L. leucocephala during the period of treatment. Therewas a 26% decrease in amount of N derived from N₂ fixation asresult of water deficit (as measured with ARA), while the decreasewas 23% when measured withboth the ¹⁵N dilution method and asaccumulated N. The three different methods for measuring N₂ fixation and effectsof water deficit on N₂ fixation are discussed. Key words: Acacia albida, ARA, Casuarina equisetifolia, Leucaena leucocephala, ¹⁵N dilution, N₂N fixation, water deficit