ALBERT

Notes: Successful long-term wetland restoration efforts require consideration of hydrology and currounding land use during the site selection process. This article describes an approach to initial site selection in the San Luis Rey River watershed in southern California that uses watershed-level information on basin topography and land cover to rank the potential suitability of all sites within a watershed for either preservation of restoration. This approach requires the use of a geographic information system (GIS)to map relative wetness and land cover within a watershed. Relative potential wetness values were derived from U.S. Geological Survey (USGS) 30-m digital elevation models by calculating the flow that would potentially accumulate at all 30-m × 30-m pixels within the watershed. Land cover was derived from a Landsat scene covering the 1500 Km2 study area. We ranked sites (contiguous groups of pixels 〉 1 ha with similar land cover) in terms of their potential for restoration or preservation based on their wetness values (Iow, medium, and high), size, and proximity to existing riparian vegetation. Sites with medium or high wetness values and extant vegetation were identified as potential preservation sites. Agiricultural or barren sites with medium to high wetness were identified as potential restoration sites. Approximately 5500 ha (3.67% of the total watershed) were prioritized for preservation or resloration.

Notes: This historical and conceptual overview of riparian ecosystem restoration discusses how riparian ecosystems have been defined, describes the hydrologic, geomorphic, and biotic processes that create and maintain riparian ecosystems of the western USA, identifies the main types of anthropogenic desturbances occurring in these ecosystems, and provides an overview of restoration methods for each disturbance type. We suggest that riparian ecosystems consist of two zones: Zone I occupies the active floodplain and is frequently inundated and Zone II extends from the active floodplain to the valley wall. Successful restoration depends n understanding the physical and biological processes that influence natural riparian ecosystems and the types of disturbance that have degraded riparian areas. Thus we recommend adopting a process-based approach for riparian restoration. Disturbances to riparian ecosystems in the western USA result from streamflow modifications by dams, reservoirs, and diversions; stream channelization; direct modification of the riparian ecosystem; and watershed disturbances. Four topics should be addressed to advance the state of science for restoration of riparian ecosys-tems: (1) interdisciplinary approaches, (2) a unified framework, (3) a better understanding of fundamental riparian ecosystem processes, and (4) restoration po-tential more closely related to disturbance type. Three issues should be considered regarding the cause of the degraded environment: (1) the location of the causative disturbance with respect to the degraded riparian area, (2) whether the disturbance is ongoing or can be elim-inated, and (3) whether or not recovery will occur nat-urally if the disturbance is removed.

Notes: A two-stage system for selecting stream reaches and riparian communities for restoration was applied to the 80-km San Luis Rey River below the Lake Henshaw dam in southern California. In the first satge, data from topographic quadrangles and aerial photographs were analyzed to define and classiy reaches. These analyses concluded that (1) 28 km of the river and adjacent floodplain were suitable for second-stage evaluation of restoration needs and (2) 32 km met criteria for reference conditions at the stream reach scale and should be protected from further impacts. The remaining 20 km of the river and floodplain were considered unsuitable for restoration to reach-scale reference conditions; individual sites may be restored under existing regulatory review. Second-stage field sampling provided data on vegetation and floodplain landforms and substrate from more thatn 3000 plots within the 28 km of river and 1120 ha of floodplain selected for further Study. Classification of floristic samples stratified by landform/substrate class indicated six primary riparian communities on the floodplain, some associated with particular floodplain landform/substrate classes and others ubiquitous. Reference conditions for these communities were interpreted from the data. There were two major departures from reference conditions: tree-dominated communities were less extensive than historic levels and exotic plants had significantly invaded some landforms and communities, displacing natural com-munities. General goals would include restoration of tree communities and removal of exotics, with further consideration of site-specific objectives. The results included estimates of the areas by community type re-quiring restoration. The approach was developed for streams in the semi-arid western United States, but it may be adapted for use elsewhere.

Notes: This study compares the results of Olson and Harris (1997) and Russell et al. (1997) in their work to prioritize sites for riparian restoration in the San Luis Rey River watershed. Olson and Harris defined reaches of the mainstem and evaluated the relative potential for restoration and protection based on cover of natural vegetation, land use, and connectivity. Then they used data on geomorphic conditions, plant species composition, and community structure to prescribe strategies for restoration. Russell et al. used a modeling approach within a geographic information system to combine data on wetness and land use/land cover to identify areas with potential for protection and restoration. They prioritized the areas based on patch size and proximity to extant riparian habitat. The main-stem and associated floodplain defined by Olson and Harris was more than twice the size of the area defined by Russell et al., because Olson and Harris considered the entire valley floor, whereas Russell et al. used a wetness index to identify saturated zones within the floodplain. For seven of the twelve management units delineated along the mainstem, the two studies agreed on a strategy of restoration or protection. They differed on two. No comparison could be made of the three units for which Olson and Harris used project review, a unique category. Agreement of the results is due to the similarity of criteria used to identify and rank sites for protection and restoration; disagreement is due primarily to the level of resolution of the data. Both approaches have potential for use in watershed-level planning. The predictive power of the two approaches may be maximized when they are used in a complementary fashion.

Notes: We present a conceptual model for identifying restoration sites for riparian wetlands and discuss its application to reaches within the Upper Arkansas River basin in Colorado. The model utilizes a Geographic Information System (GIS) to analyze a variety of spatial data useful in characterizing geomorphology, hydrology, and vegetation of riparian wetland sites. The model focuses on three basic properties of riparian wetland sites: relative soil moisture, disturbance regime, and vegetative characteristics. A relative wetness index is used to define nominal soil moisture classes within the watershed. These classes generally coincide with uplands (low), channel margins (moderate), and channels or open water (high). Vegetative conditions are characterized using color infrared aerial photographs. Land cover types are grouped into five major land cover classes: riparian, moist herbaceous, bare ground, upland, and open water. Disturbance regime is characterized by a reach-based index of specific power (ω). Preliminary results indicate that reaches within the Upper Arkansas River basin can be classified as high energy (ω≥ 8 W/m2) or low energy (ω≤ 3W/m2), using discharge estimates that reflect the 10-year flood event. Field surveys of channel and floodplain conditions show that high-energy reaches (ω≥ 8 W/m2) are characterized by sites where the channel occupies a large proportion of the valley bottom. By contrast, low-energy reaches (ω≤ 3 W/m2) are characterized by meandering channels with wide alluvial valleys. Restoration potential is evaluated as a combination of nominal scores from wetness, land cover, and disturbance indices. Application of these methods to field sites within the Upper Arkansas River basin identifies a wide range of riparian wetland sites for preservation or restoration. Potential sites within identified reaches are prioritized using size and proximity criteria.

Notes: A major goal of population biologists involved in restoration work is to restore populations to a level that will allow them to persist over the long term within a dynamic landscape and include the ability to undergo adaptive evolutionary change. We discuss five research areas of particular importance to restoration biology that offer potentially unique opportunities to couple basic research with the practical needs of restorationists. The five research areas are: (1) the influence of numbers of individuals and genetic variation in the initial population on population colonization, establishment, growth, and evolutionary potential; (2) the role of local adaptation and life history traits in the success of restored populations; (3) the influence of the spatial arrangement of landscape elements on metapopulation dynamics and population processes such as migration; (4) the effects of genetic drift, gene flow, and selection on population persistence within an often accelerated, successional time frame; and (5) the influence of interspecific interactions on population dynamics and community development. We also provide a sample of practical problems faced by practitioners, each of which encompasses one or more of the research areas discussed, and that may be solved by addressing fundamental research questions.

Notes: Cool smoke treatments were applied to unmined Eucalyptus marginata (jarrah) forest soils, rehabilitated bauxite mine soils, and broadcast seed to determine if enhancement in germination could be effected with a view toward maximizng the establishment of species in bauxite mines in Western Australia.Forest sites showed a 48-fold increase in total germinants from the soil seed bank when treated with aerosol smoke. Newly returned bauxite mine soils showed a greater than threefold increase in total germinants after the same treatment. There were also significant increases in the number of species germinating in response to the aerosol smoke treatment in both the forest and the mined soils. Similarly, application of smoked water to the soil seed bank in previously mined sites elicited a significant positive germination response, increasing total germinants and species numbers by 56 and 33%, respectively.Treatment of mixed seed lots with aerosol smoke before broadcast resulted in highly significant improvement in germination when compared to untreated seed. Both total number of germinants, and number of species emerging from mined sites were positively influenced (85% and 34% increases, respectively).Ten target species were used to determine the relative effectiveness of different methods of smoke treatment on the germination of broadcast seed. Nine of the species involved displayed a promotive effect with at least two of the treatments. Generally, however, aerosol smoking of seed before broadcast proved to be the more effective approach. As a result of these findings, all broadcast seed for use in Alcoa's bauxite mined areas in the southwest of Western Australia is now routinely smoke treated before application.

Notes: Understanding Cladium jamaicense (sawgrass) seedling establishment is an important component of an Everglades restoration program because the degree of sawgrass recovery and concurrent Typha domingensis (cattail) decline will be used to evaluate restoration success. To understand sawgrass recovery at locations with increased soil nutrients, we tested the effects of transplanting sawgrass seedlings to pots at different densities and investigated how nutrient additions affect seedling growth. Survivorship of seedlings transplanted into moist commercial potting soil at three densities ranged from 61% to 95%. After 6 months, maximum survivorship (90%) occurred at medium densities (2–4 seedlings per pot 16 cm in diameter). Nutrient additions, totaling 6.5 N g/m2, 9.8 P g/m2, 6.5 g/m2, were applied approximately 4 months after seedlings were transplanted. The biomass of the plants receiving nutrient additions (pulsed) was significantly higher (by over 30%) than plants with no nutrient addition (control). Photosynthetic rates for nutrient-enriched plants (measured 6-weeks after the nutrient additions) were significantly greater (by 32–45%) than for control plants. Instantaneous leaf water use efficiency increased significantly (by more than 20%) in pulsed plants. The results suggest that preventing root damage is crucial for the success of trans planted sawgrass seedlings and that nutrient additions enhanced seedling growth.

Notes: I evaluated responses by 16 native woody species to differential soil compaction and density of ground cover. The trees and shrubs studied represent sites in southern Illinois that commonly have restrictions to root growth from soil or drainage conditions. The study site was a restored surface coal mine in southern Illinois with a rooting medium compacted by grading and a dense ground cover of pasture species. Soil compaction was alleviated in half the study area before tree planting by mechanically ripping the soil to a depth of 1.2 m. Roots of half the trees and shrubs were dipped in a Terra® slurry before planting, and the ground cover around all planting spots was afterwards sprayed with herbicide. In year 2 after planting the ground cover in half of the unripped and half of the ripped area was further controlled by repeated application of herbicides. Ripping significantly increased height growth of all trees combined and all species individually in each year of the study. Second-year control of ground cover increased height growth of all trees combined and of seven species individually. Some species were damaged by herbicides. Terra® had little evident effect on species performance. Animal damage reduced early survival and growth, especially of Acer (maple) and Cornus (dogwood) species, and later growth of Quercus rubra (red oak). Removal of ground cover with herbicides tended to increase deer browse. Soil ripping, herbicide application, and choosing tree species unattractive to deer can be recommended to increase success in planting trees for forest restoration.

Notes: Woodlands dominated by Eucalyptus spp. in temperate southeastern and southwestern Australia have been extensively cleared for agriculture and are often badly degraded by livestock grazing. This has resulted in the loss of biodiversity and widespread land degradation. The continuing decline of these woodlands has become a concern for the conservation of biodiversity, and there is a growing interest among farmers, land managers, and researchers in developing techniques for restoring them. Currently few scientific guidelines exist for undertaking woodland restoration programs. We use a state and transition model to develop hypotheses on restoration strategies for salmon gum (Eucalyptus salmonophloia) woodlands. We consider that this approach provides a suitable framework for organizing knowledge and identifying areas where further information is needed, and hence provides a useful starting point for a restoration program. The model has the potential to provide a tool for land managers with which they can assess the action and effort needed to undertake woodland restoration in agricultural landscapes.

Notes: Most of the world's forest has been cleared, cultivated, and then often abandoned. In many instances these areas have changed to successionally arrested grasslands, shrublands, or fernlands maintained by frequent fires and high herbivore populations. Many studies have shown that various herbaceous, nitrogen-fixing legumes can protect soil surfaces, retain soil moisture, improve soil fertility, and retard ground fires. Our objective was to ascertain if some of these species can potentially inhibit herbivory and satisfactorily establish in these arrested grassland areas to serve as sites for reforestation. We evaluated the potential for four species of nitrogen-fixing legumes (Calapogonium mucunoides, Centrosema pubescens, Desmodium ovalifolium, and Pueraria phaseoloides) to establish on exposed soil within successionally arrested grasslands of Panicum maximum and Cymbopogon nardus in the central hills of Sri Lanka. Four different sites within rectangular grassland areas were cleared of graminoids and sown with seed of each legume. Half of each clearing was protected from browsing rabbits and porcupines, and half was not protected. After 6 months, certain plots were destructively sampled to determine dry biomass gain for each species and treatment. Analyses of variance were performed to test for differences among sites, treatments, and species. All three factors revealed differences, indicating that species must be matched to site. On sites with high amounts of herbivory, D. ovalifolium had the greatest dry biomass gain after 6 months of growth, possibly because of its relatively low nitrogen and moisture content. Where herbivory was absent, P. phaseoloides and C. muconoides had the greatest dry biomass gain. Dry biomass gain of all four legume ground covers was low on sites with lowest pH and nutrient concentrations. Under conditions of low relative fertility and low pH, establishment of the tested legumes failed. Though soil moisture availability was not measured, we speculate that these low fertility sites were also prone to drought. Findings support the site-specific establishment of legume species for purposes of reforestation and watershed protection in central Sri Lanka. This work is applicable to other regions particularly dominated by successionally arrested grasslands with similar circumstances in other parts of south and southeast Asia.

Notes: Oyster cultch was added to the lower intertidal fringe of three created Spartina alterniflora marshes to examine its value in protecting the marsh from erosion. Twelve 5-m-wide plots were established at each site, with six randomly selected plots unaltered (non-cultched) and cultch added to the remaining (cultched) plots. Within each cultched plot, cultch was placed along the low tide fringe of the marsh during July 1992, in a band 1.5 m wide by 0.25 m deep. Marsh-edge vegetation stability and sediment erosion were measured for each plot from September 1992 to April 1994. Significant differences (p 〈 0.05) in marsh-edge vegetation change were detected at the only south-facing site after a major southwester storm. Significantly different rates of sediment erosion and accretion also were observed at this same site. Areas upland of the marsh edge in the cultched areas showed an average accretion of 6.3 cm, while noncultched treatment areas showed an average loss of 3.2 cm. A second site, with a northern orientation, also experienced differential sediment accretion and erosion between treatment type, caused instead by boat wakes that were magnified by the abutment of a dredge effluent pipe across the entire front fringe of the site. During this period we observed significant differences in sediment accumulation, with the areas upland of the marsh edge in the cultched treatment having an average accretion of 2.9 cm and the noncultched an average loss of 1.3 cm.

Notes: Books reviewed in this article: The Significance and Regulation of Soil Biodiversity. Harold P. Collins, G. Philip Robertson, and Michael J. Klug, editors

Notes: Declines in native plant and animal communities have prompted new interest in the restoration of aquatic and riparian ecosystems. Past restoration activities typically have been site specific, with little thought to processes operating at larger scales. A watershed analysis process developed in the Pacific northwest identifies four operating scales useful in developing restoration priorities: region, basin, water-shed, and specific site. Wtershed analysis provides a template for restoration practitioners to use in prioritizing restoration activities. The template identifies seven key steps necessary to understand and develop restoration priorities: (1) characterization, (2) identification of key issues and questions, (3) documentation of current conditions, (4) description of reference conditions, (5) identification of objectives, (6) summary of conditions and determination of causes, and (7) recommendations. When a similar process was used in the Uinta Mountains, Utah, and in the Sluslaw National Forest, Orgon, specialists were able to identify key habitat conditions and habitat forming processes and then to establish restoration priorities and implement the appropriate activities. Watershed analysis provides a valuable set of tools for identifying restoration activities and is currently being used throughout the Pacific Northwest to develop management strategies and restoration priorities. Although the analysis requires significant time, money, and personnel, experience suggests that watershed analysis provides valuable direction for managing aquatic and riparian resources.

Notes: Landscape ecology focuses on questions typically addressed over broad spatial scales. A landscape approach embraces spatial heterogeneity, consisting of a number of ecosystems and/or landscape structures of different types, as a central theme. Such studies may aid restoration efforts in a variety of ways, including (1) provision of better guidance for selecting reference sites and establishing project goals and (2) suggestions for appropriate spatial configurations of restored elements to facilitate recruitment of flora/fauna. Likewise, restoration efforts may assist landscape–level studies, given that restored habitats, possessing various patch arrangements or being established among landscapes of varying diversity and conditions of human alteration, can provide extraordinary opportunities for experimentation over a large spatial scale. Restoration studies can facilitate the rate of information gathering for expected changes in natural landscapes for which introduction of landscape elements may be relatively slow. Moreover, data collected from restoration studies can assist in validation of dynamic models of current interest in landscape ecology. We suggest that restoration and landscape ecology have an unexplored mutualistic relationship that could enhance research and application of both disciplines.

Notes: Restoration ecologists use reference information to define restoration goals, determine the restoration potential of sites, and evaluate the success of restoration efforts. Basic to the selection and use of reference information is the need to understand temporal and spatial variation in nature. This is a challenging task: variation is likely to be scale dependent; ecosystems vary in complex ways at several spatial and temporal scales; and there is an important interaction between spatial and temporal variation. The two most common forms of reference information are historical data from the site to be restored and contemporary data from reference sites (sites chosen as good analogs of the site to be restored). Among the problems of historical data are unmeasured factors that confound the interpretation of historical changes observed. Among the problems of individual reference sites is the difficulty of finding or proving a close match in all relevant ecological dimensions. Approximating and understanding ecological variation will require multiple sources of information. Restoration, by its inherently experimental nature, can further the understanding of the distribution, causes, and functions of nature's variation.

Notes: Books reviewed in this article: Water Quality: Management of a Natural Resource James Perry and Elizabeth Vanderklein

Notes: The U.S. Bureau of Reclamation created a shallow, 110-m channel to provide habitat for two endangered fishes, Cyprinodon elegans (Comanche Springs pupfish) and Gambusia nobilis (Pecos gambusia), at the site of the fishes' former natural habitat. The ciénega (marsh) associated with Phantom Lake Spring in Jeff Davis County, Texas, was destroyed by the creation of an irrigation canal system. In 1993, the endangered fishes were stocked into the refuge with individuals from the irrigation canals, and in the case of C. elegans, hatchery stocks. The condition of habitat, status of fish populations, and fish ecology within the refuge were then monitored for two years. The abundance and density of both species increased in accordance with aquatic plant development. Cyprinodon elegans abundance peaked after one year and stabilized at an average density of 14.7/m2 by the end of our study. Juvenile C. elegans were always rare, which may indicate that the population reached the refuge's carrying capacity and that recruitment is low. Gambusia nobilis was the most abundant fish in the refuge (average density 96/m2), used the entire refuge, and outcom-peted nonindigenous G. geiseri. The two Gambusia species used similar habitats but showed almost no dietary overlap. High densities of aquatic plants reduced the amount of open water areas necessary for C. elegans. The refuge will sustain the two endangered fishes at this historic site of endemism while maintaining flow to the irrigation system; however, the refuge is not equivalent to a restored ciénega.

Notes: No guidelines are currently available that advise which soil properties of arable land can be used to suggest suitable locations for heathland reconstruction. This paper reviews studies comparing soil properties of heathland or semi-natural grassland with those of adjacent arable fields, investigations in the autecology of the dominant heathland plant, Calluna vulgaris (common heather), and long-term experiments of fertilizer inputs on arable soils. Three properties must be assessed before the suitability of a field can be determined: extractable phosphorus, exchangeable calcium, and pH. A number of other nutrients may also be important, but evidence is currently insufficient to substantiate this. Natural changes in levels of extractable phosphorus, exchangeable calcium, and pH appear to be very slow, so nutrient stripping and acidification will be necessary where recommended levels are exceeded to successfully restore heathland vegetation.

Notes: Ants are widely used as bioindicators in Australian land assessment and monitoring programs, particularly in relation to ecosystem restoration following mining. Little is known, however, about the relationship between ant community development and key ecological processes such as nutrient cycling. We have examined the relationship between ant species richness and soil microbial biomass at 17 sites subject to disturbance by mining in the Kakadu region of Australia's Northern Territory. The number of ant species recorded ranged from 7 at an unvegetated site undergoing restoration to 43 at a site that was undisturbed except for edge effects. Soil microbial biomass ranged from 19.3 to 134.3 μgC/g. Ant species richness was positively correlated with soil microbial biomass (r= 0.638), more so than was plant species richness (r= 0.342 for total plant species, r= 0.499 for woody species only). Our findings demonstrate a correlation between aboveground ant activity and belowground decomposition processes at disturbed sites, thereby providing support for the use of ants as indicators of restoration success following disturbance. Interestingly, when a range of undisturbed sites in the region was considered, a negative rather than positive relationship between ant richness and soil microbial biomass was found. This illustrates the importance of distinguishing between variation within a habitat due to disturbance and variation across different habitats when searching for indicators of ecological change.

Notes: Bottomland hardwood forests in the southcentral United States have been cleared extensively for agriculture, and many of the remaining forests are fragmented and degraded. During the last decade, however, approximately 75,000 ha of land—mainly agricultural fields—have been replanted or contracted for replanting, with many more acres likely to be reforested in the near future. The approach used in most reforestation projects to date has been to plant one to three overstory tree species, usually Quercus spp. (oaks), and to rely on natural dispersal for the establishment of other woody species. I critique this practice by two means. First, a brief literature review demonstrates that moderately high woody species diversity occurs in natural bottomland hardwood forests in the region. This review, which relates diversity to site characteristics, serves as a basis for comparison with stands established by means of current reforestation practices. Second, I reevaluate data on the invasion of woody species from an earlier study of 10 reforestation projects in Mississippi, with the goal of assessing the likelihood that stands with high woody species diversity will develop. I show that natural invasion cannot always be counted on to produce a diverse stand, particularly on sites more than about 60 m from an existing forest edge. I then make several recommendations for altering current reforestation practices in order to establish stands with greater woody species diversity, a more natural appearance, and a more positive environmental impact at scales larger than individual sites.

Notes: We evaluated the ecological and reproductive characteristics of plant species occurring in severely disturbed areas that were revegetated with exotic grasses. We identified those species with the best combination of attributes that increase their probability of success in degraded lands. Fifteen degraded areas were studied in two different bioclimatic regions, a high premontane humid bioclimate and a low premontane humid bioclimate. The frequency of native colonizing species and the presence of arbuscular mycorrhizal colonization in their roots were evaluated. The sexual and breeding system, pollination mode, fruit set, and dispersal syndrome of ten of the most frequent colonizing species were also studied. The floristic survey of the colonizing species revealed a similarity to the reported flora of the treeless savannas that are dominant in the region. Bioclimatic conditions prevailing in the degraded lands seem to be an important factor for the presence of colonizing species and for species richness. All colonizing species studied were mycorrhizal, and for this reason the restoration program in these degraded areas should take mycorrhizae into account, reintroducing them or manipulating the soils to increase the mycorrhizal inoculum. We suggest Scleria cyperina and Trachypogon plumosus to start or promote the natural succession in the degraded areas from La Gran Sabana. Because their frequency is high and their reproductive system is less dependent on biotic factors, these species stand out in the studied areas.

Notes: Disturbance of coastal sage scrub in southern California has led to extensive displacement of native shrubs by exotic annual grasses. The initial conversion from shrubland to exotic grassland is typically associated with disturbance caused by intense grazing, high fire frequency, or mechanical vegetation removal. While native shrubs have been shown to recolonize annual grasslands under some conditions, other annual grasslands are persistent and show no evidence of shrub recolonization. This study examined the mechanisms by which annual grasses may exclude native shrubs and persist after release from disturbance. Grass density was manipulated in experimental plots to achieve a series of prescribed densities. Artemisia californica, a dominant native shrub, was seeded or planted into the plots and responses to the grass density treatments were measured over two growing seasons. A. californica germination, first season growth, and survival were all negatively related to the density of neighboring annual grasses. The most probable mechanism underlying the reduction of first season growth and survival was depletion of soil water by the grasses. The effects of the grasses on A. californica were no longer significant in the second season. The results of this study indicate that Mediterranean annual grasses reduce recruitment and can persist by inhibiting post-disturbance establishment of A. californica from seed. Although succession alone may not return disturbed annual grasslands to their former shrubland composition, the results suggest that restoration can be achieved by using container plantings or grass removal followed by seeding.

Notes: This study evaluated the genetic consequences of a reintroduction of the endangered annual plant Cordylanthus maritimus ssp. maritimus to Sweetwater Marsh (San Diego County, California). A survey of 21 enzyme loci in natural populations revealed that genetic diversity is very low and is primarily found as rare alleles at a few loci, making this species especially susceptible to the loss of alleles and heterozygosity through genetic drift. The reintroduction was performed in 1991 and 1992 by sowing seeds (collected from Tijuana Estuary) in numerous small patches of suitable habitat. For this study, leaf tissue was collected from all plants in all patches during flowering in 1995 and surveyed for genotype at the three enzyme loci that are polymorphic at Tijuana Estuary. Rare alleles were absent in 27 out of 30 patches for Pgm-1, in 17 out of 30 patches for Pgm-2, and in 10 out of 11 patches for Mdh-1. In all, half of the patches lacked any rare allele. Rare alleles tended to occur in patches with few individuals. Overall rare allele frequency was lower than in the colonies from which seeds were collected at two of the three loci, and heterozygosity was reduced. The Sweetwater Marsh population is at risk of losing most of its genetic variation at enzyme loci through the extinction of patches with few individuals. Future reintroduction attempts should attempt to create contiguous sets of patches or to periodically reseed existing patches to reduce the loss of genetic variation.

Notes: Books reviewed in this article: Riparian Ecosystem Recovery in Arid Lands: Strategies and References. Mark K. Briggs Land Restoration and Reclamation: Principles and Practice James A. Harris, Paul Birch, and John Palmer

Notes: Effects of waterlogging were studied in the field and under glasshouse conditions on two clonal lines of Eucalyptus camaldulensis Dehnh (river red gum), which are used in the rehabilitation of damaged agricultural catchments in Western Australia. The plantation of 9-year-old trees was in a position that covered a range of waterlogging and salinity conditions. Up-slope the water table was deeper (0.65–1.5 m), whereas the water table was closer to the ground surface down-slope (0.45 m in winter; 1.25 m in summer). Salinity was greater downslope and increased at the end of the dry summer, remaining high until diluted by the winter rains. Trees of both clonal lines were smaller downslope and used less water over the year. Clone M80 used more water in winter; clone M66 more in summer. In the field, the roots of clone M80 were evenly distributed through the soil profile, while roots of clone M66 decreased with increasing depth. Production of new root terminals varied with season. Greatest production was in spring and early summer, with much lower production over late autumn and winter. Only clone M66 produced new root terminals at depth (60–75 cm) during the drier months of late summer and early autumn. At this time, saline ground-water was the main source for water uptake. To explore clonal differences more closely, the effects of prolonged waterlogging were studied under glasshouse conditions. Clone M80 grew similarly under freely drained and continuously waterlogged conditions for the experimental period (21 weeks). The response under continuously waterlogged conditions was achieved through adventitious root production. By contrast, growth of clone M66 was suppressed under continuous waterlogging, a response associated with the lack of adventitious root production. The results from field and glasshouse studies suggest that clone M80 is more adapted to waterlogging by relatively fresh water than clone M66, but that clone M66 may use water of higher salinity than clone M80. Clone M80 would be better suited to higher positions in partially cleared catchments, where rainfall provides relatively fresh soil water. Clone M66 is better suited to lower catchment positions due to its ability to utilize more saline groundwater. Restoration of the water balance of damaged agricultural catchments can be best managed by matching specialized genotypes with particular catchment positions.

Notes: I conducted small-mammal trapping surveys on a desert scrub restoration site in Palm Springs, California, to document concomitant recovery of the rodent community. These surveys were conducted following quantitative vegetation sampling efforts that indicated that a predefined successful restoration criterion of 15% total shrub cover had been met throughout most of the area. But shrub cover, native shrub cover, herb cover, native herb cover, total cover, and total native cover remained significantly lower in the restoration area than in undeveloped desert scrub immediately surrounding the site. Native herb species richness was also generally lower in the restoration area. Despite these vegetation differences, rodent diversity, evenness, and abundance were very similar between the restoration and natural areas (they were consistently slightly higher in the restoration area). More diverse microhabitats, proximity to water, and reduced competition with harvester ants may have contributed to this outcome. If ecosystem restoration is the goal, reestablishment of a faunal community in restored habitat, rather than surpassing a predefined percent cover of vegetation, may be a better indicator of success, because plant cover proved to be a poor predictor of mammal success.

Notes: Books reviewed in this article: California Rivers and Streams: The Conflict Between Fluvial Processes and Land Use. Jeffery F. Mount Evolution and the Aquatic Ecosystem: Defining Unique Units in Population Conservation. J. L. Nielson, editor

Notes: Closed or abandoned landfills represent significant land areas, often in or near urban centers, that are potential sites for ecological restoration of native woodlands. But current guidelines in many jurisdictions do not allow for the installation of trees or shrubs above landfill clay caps, although these plants have many environmental, functional, and aesthetic advantages, including a rapid start to community succession. Typical closure procedures for capped landfills include only a grass cover to control moisture infiltration and impede soil erosion. The main concern that limits the application of a woody cover to a closed landfill is that roots may penetrate and weaken the clay cap. As part of a comprehensive experimental program on woodland restoration, we installed 22 tree and shrub species on Staten Island, New York (the Fresh Kills Sanitary Landfill). We found no evidence that roots of the transplanted woody plants penetrate caps used on these landfills. Root growth requirements and dynamics stop penetration of these materials. Anoxic and acidic conditions were found in the sandy subsoil above the cap, as indicated by corrosion patterns on steel test rods. Also, the intensity of mycorrhizal infection on the experimental plants was high in the surface soil and decreased progressively with increasing soil depth. The potential vertical rooting depth during this time period was greater than that occurring over the clay cap. This was shown from data collected on a nearby control site, where seven of the species were installed on an engineered soil lacking a clay barrier layer, and roots of all seven species penetrated deeper than on the landfill. The engineered landfill soils are poor growth media for roots, and below ground constraints that limit restoration on these sites must be addressed.

Notes: The Northern Rocky Mountain Wolf Recovery plan proposed reintroduction of Canis lupus (gray wolf) to Yellowstone National Park and central Idaho as part of a wolf restoration plan for the northern Rocky Mountains of the United States. Strong opposition from some factions within the region forestalled the action for two decades. An environmental impact statement, conducted in 1992–1994 with extensive public input, culminated in a proposal to reintroduce wolves designated as “non-essential—experimental” under Section 10 (j) of the federal Endangered Species Act. This approach, approved by the Secretary of the Interior in 1994, provided for wolf restoration while allowing management flexibility to deal with concerns of the local public. A reintroduction plan was developed in the summer and fall of 1994. Acquiring, holding, transporting, and releasing suitable wolves for reintroduction presented a myriad of technical and logistical challenges that required effective planning and coordination by all participants. In January 1995, 29 wolves were captured in Alberta and transported to Yellowstone National Park (14) and central Idaho (15). Idaho wolves were freed immediately upon arrival; Yellowstone wolves (three family groups) were held in acclimation pens in the park until late March. Most Idaho wolves traveled extensively within the area intended for them, averaging 82 km net distance away from release sites after 5 months (range = 30–220 km), and three male-female pairs formed by July. After 5 months in the wild, at least 13 of 15 Idaho-released wolves were alive within the intended area, as were 13 of 14 Yellowstone wolves; one wolf was known to have been illegally killed in each area. No livestock were killed. Wolves released into Yellowstone Park continued to live as packs, stayed closer to their release sites (x = 22 km at end of June), and settled into home ranges; two packs produced a total of nine pups. The progress of the reintroduction program in its first year far exceeded expectations. Reintroductions of about 15 wolves to each area for 2–4 more years are scheduled, but the project may be shortened because of early successes. Future reintroduction planners can expect sociocultural issues to pervade the effort, but they can be optimistic that, from a biological standpoint, reintroduction of wolves has strong potential as a restoration technique.

Notes: This paper summarizes the accumulation of nutrient capital during the natural afforestation of iron-smelting slag. The study site was the Beckley Furnace (North Caanan, Connecticut, U.S.A.) slag dump, which was abandoned around 1918. Currently, a Pinus strobus (white pine)-dominated forest occupies the site. Our primary objective was to determine the relationships among stand development and belowground nutrient pools to evaluate controls on the rates at which nitrogen, calcium, magnesium, and potassium accumulate in the developing forest. Along a developmental gradient of increasing stand basal area, we measured (1) aboveground biomass, (2) root biomass, (3) above-ground nutrient capital, (4) belowground nutrient capital, (5) soil weight, (6) soil organic carbon weight, (7) soil moisture, and (8) understory richness and density. Regression analysis suggests, as expected, that the addition of organic matter controls the accumulation of above- and belowground nutrients, soil water, and understory plant composition and distribution. The rather rapid rate of nitrogen accumulation suggests that allochthonous as well as autochthonous organic matter is an important source of nutrients for the developing forest soil and vegetation. Compared to those of other northeastern forests, soils in the most developed areas at the Beckley site have accumulated more than enough labile nutrient capital in 75 years to support a forest typical of the region. In the most developed stands, understory composition indicates mesic soil conditions, and that within the next 75 years or so the Beckley slag dump will be floristically very similar to the surrounding forest.

Notes: Extensive land loss, which is mostly wetland loss, has taken place during this century in the Mississippi River delta and other river deltas. Our purpose was to evaluate the effectiveness of constructing “artificial” crevasses, or cuts in the natural levee, made by the U.S. Fish and Wildlife Service in the Delta National Wildlife Refuge (DNWR) to slow or reverse this type of land loss. Land growth of the crevasses was determined from aerial photographs and was related to crevasse-site characteristics. The newly constructed crevasses create emergent wetlands after 2 years of subaqueous growth at about 4.7 ha/year and an average cost of $21,377 per crevasse. The present total cost per hectare declines with age as new land builds, and it will equal $48 per hectare if all the open water in the receiving ponds fills in. At these rates, the net land loss rates in the DNWR measured from 1958 to 1978 would be compensated for by the building of 63 crevasses, 24 of which are already in place.

Notes: This paper describes an ecosystem approach (the Bradbury framework) to prioritizing watersheds for watershed restoration and salmon recovery, and gives an example of its application. The framework was applied at three spatial scales (in descending urder) to prioritize (1) river basins within the north coast geographic area of Oregon (USA), (2) watersheds within the Tillamook Bay basin, and (3) restoration activities at the watershed level, Implementing the framework identified the Nehalem and Tillamook Bay basins as high priority for the north coast of Oregon. Within the Tillamook Bay basin, the Wilson, Kilchis, and Trask river watersheds emerged as high priority. Preliminary analysis indicated that controlling sediment sources by addressing upland road conditions and allowing floodplain and riparian ecosystems to recover are highest priority protection and restoration activities within the Tillamook Bay basin. The sample application demonstrates that an ecosystem approach (the Bradbury framework) is particularly advantageous where data are limited, although previous identification of relatively intact areas is required. Implementing the framework is intended to lead to restoration of native species, but it may not provide immediate assistance for some species or populations of concern.

Notes: This paper describes a two-stage system for prioritizing stream reaches and riparian communities along a given river for protection or restoration. The system uses associations between geomorphology and riparian vegetation at stream reach and community scales as a basis for defining reference conditions. First-stage reach classification involves collecting and analyzing data from topographic maps and aerial photographs. These data, along with judgment-based criteria for ranking reaches relative to reference conditions, are used to classify stream reaches as suitable for protection, recommended for mitigation or restoration within existing site-specific regulatory procedures, or requiring further analysis to evaluate community-scale restoration needs. Second-stage field sampling is conducted on the reaches needing further analysis to determine the riparian communities present, the associations between communities and floodplain landforms, and reference community conditions. This stage requires collection of fields data on geomorphic conditions, plant species composition, and plant community structure. Cluster analysis or a comparable technique is used to classify plant communities associated with floodplain landforms and identify reference conditions for each landform. Community structure and species compositiotion are compared to reference conditions to define restoration possibilities at the community scale. The combined results from stream reach and community scale analysis provide a strategy for protecting and restoring riparian resources for a whole river. Implementation requires further site-specific information on hydrology, geomorphology, and other factors.

Notes: In this study, we examined the site-specific effects of a large episodic flood on the riparian vegetation within the floodplain of the San Luis Rey River in southern California. Using multispectral airborne videography, we quantified percent cover of riparian vegetation, cultivated agricultural land, urban surfaces, upland vegetation, bare soil, and water within 22 sections of flood-plain, both before and after a large flood (January 13, 1993). We also quantified the amount of these cover types within bands of the watershed 1 km wide × 5 km long directly upstream of each floodplain site. The amount of riparian vegetation destroyed by the flood within each section varied from nearly zero to almost 40% of pre-flood coverage. The magnitude of loss in riparian vegetation was most strongly related to the amount of riparian vegetation initially present in the floodplain and the amount of urban surfaces in the nearby watershed. These results suggest that riparian vegetation within the San Luis Rey River floodplain is generally at high risk of destruction from large floods, and that this risk is exaggerated in areas with high urban development. We infer from these results that sites near existing large areas of intact riparian vegetation and away from urban development will have the highest potential for successful long-term restoration.

Notes: Airborne multispectral videography has become a useful tool for classifying and mapping riparian vegetation because its spectral and digital format has high spatial resolution. This paper explains the processing steps and procedures required to use this type of spectral imagery for riparian vegetation classification and mapping. A description of the Utah State University airborne system is given, along with the image processing steps used for developing finalized products, because these are used in other articles in this volume. Examples of riparian applications using high-resolution imagery are included.

Notes: Nine Articles in the special issue of Restoration Ecology addrssing the subject of site selection for riparian restoration activities were critically examined for this review. The approaches described make significant and original contributions to the field of riparian restoration. All are interdisciplinary to some extent, often combining the fields of hydrology, geomorphology, and biology in the design of restorationss. A common component among the articles is that they take a broad view, if not a watershed view, of restoration site selection. The approaches can be generally descrobed as top-down strategic approaches to siting restorations, as opposed to the more methods- and site-driven bottom-up, or tactical, approach. All the articles recognize the importance of developing endpoints related to the ecological function of riparian ecosystems. they succeed in their quest for these indicators of ecological function to varying degrees. The most common indica-for used in these papers is riparian vegetation. Several additional elements of scientific investigation, if successfully pursued, could provide vital information and advance our understanding of riparian restoration: developing interdisciplinary approaches more fully; defining endpoints and reference conditions; implementing multiple scale approaches; viewing restorations as experimental ecosystem manipulations; developing a philosophy regarding exotic species; incorporating geographic information systems more often; and integrating science, society, and politics. the foundation provided by the contributions in this issue should provide a strong basis for the rapid advancement of future research in the area of riparian restoration.

Notes: Successional models are used to predict how restoration projects will achieve their goals. These models have been developed on different spatial and temporal scales and consequently emphasize different types of dynamics. This paper focuses on the restoration goal of self-sustainability, but only in the context of a long-term goal. Because of the temporal scale of this goal, we must consider the impact of processes arising outside of the restoration site as of greater importance than restoration itself. Because ecological systems are open, restoration sites will be subjected to many external influential processes. Depending on the landscape context, the impact of these processes may not be noticeable, or, at the other extreme, they may prevent the achievement of restoration objectives. A second issue is to emphasize the nature of processes in the long term, that they are a complex of characteristics such as magnitude, frequency, and extent. Ecological systems are only adapted to a range of values in each of these characteristics. Restoration often combines goals that are of different scales. Models appropriate to these goals need consideration.

Notes: The ecosystem perspective provides a framework within which most other aspects of the ecology of restoration can be incorporated. By considering the ecosystem functions of a restoration project, the restorationist is forced to consider the placement of the project in the landscape—its boundaries, its connections or lack thereof to adjoining ecosystems, and its receipts and losses of materials and energy from its physical surroundings. These characteristics may set limits on the kind(s) of biotic communities that can be created on the site. The ecosystem perspective also gives restorationists conceptual tools for structuring and evaluating restorations. These include the mass balance approach to nutrient, pollutant, and energy budgets; subsidy/stress effects of inputs; food web architecture; feedback among ecosystem components; efficiency of nutrient transfers, primary productivity and decomposition as system-determining rates; and disturbance regimes. However, there are many uncertainties concerning these concepts, their relation to each other, and their relationships to population- and community-level phenomena. The nature of restoration projects provides a unique opportunity for research on these problems; the large spatial scale of restorations and the freedom to manipulate species, soil, water, and even the landscape could allow ecosystem-level experiments to be conducted that could not be performed otherwise.

Notes: Community ecological theory may play an important role in the development of a science of restoration ecology. Not only will the practice of restoration benefit from an increased focus on theory, but basic research in community ecology will also benefit. We pose several major thematic questions that are relevant to restoration from the perspective of community ecological theory and, for each, identify specific areas that are in critical need of further research to advance the science of restoration ecology. We ask, what are appropriate restoration endpoints from a community ecology perspective? The problem of measuring restoration at the community level, particularly given the high amount of variability inherent in most natural communities, is not easy, and may require a focus on restoration of community function (e.g., trophic structure) rather than a focus on the restoration of particular species. We ask, what are the benefits and limitations of using species composition or biodiversity measures as endpoints in restoration ecology? Since reestablishing all native species may rarely be possible, research is needed on the relationship between species richness and community stability of restored sites and on functional redundancy among species in regional colonist “pools.” Efforts targeted at restoring system function must take into account the role of individual species, particularly if some species play a disproportionate role in processing material or are strong interactors. We ask, is restoration of habitat a sufficient approach to reestablish species and function? Many untested assumptions concerning the relationship between physical habitat structure and restoration ecology are being made in practical restoration efforts. We need rigorous testing of these assumptions, particularly to determine how generally they apply to different taxa and habitats. We ask, to what extent can empirical and theoretical work on community succession and dispersal contribute to restoration ecology? We distinguish systems in which succession theory may be broadly applicable from those in which it is probably not. If community development is highly predictable, it may be feasible to manipulate natural succession processes to accelerate restoration. We close by stressing that the science of restoration ecology is so intertwined with basic ecological theory that practical restoration efforts should rely heavily on what is known from theoretical and empirical research on how communities develop and are structured over time.

Notes: Conceptual and logistical challenges associated with the design and analysis of ecological restoration experiments are often viewed as being insurmountable, thereby limiting the potential value of restoration experiments as tests of ecological theory. Such research constraints are, however, not unique within the environmental sciences. Numerous natural and anthropogenic disturbances represent unplanned, uncontrollable events that cannot be replicated or studied using traditional experimental approaches and statistical analyses. A broad mix of appropriate research approaches (e.g., long-term studies, large-scale comparative studies, space-for-time substitution, modeling, and focused experimentation) and analytical tools (e.g., observational, spatial, and temporal statistics) are available and required to advance restoration ecology as a scientific discipline. In this article, research design and analytical options are described and assessed in relation to their applicability to restoration ecology. Significant research benefits may be derived from explicitly defining conceptual models and presuppositions, developing multiple working hypotheses, and developing and archiving high-quality data and metadata. Flexibility in research approaches and statistical analyses, high-quality databases, and new sampling approaches that support research at broader spatial and temporal scales are critical for enhancing ecological understanding and supporting further development of restoration ecology as a scientific discipline.

Notes: Along rural roadsides of the Sacramento Valley of California, we seeded native and non-native perennial grasses to gauge their potential value in roadside vegetation management programs. In trial I (polycultures), three seeded complexes and a control (resident vegetation only) were tested. Each seeded plant complex included a different mix of perennial grasses seeded into each of several roadside topographic zones. The seeded levels of plant complex were: native perennial grasses 1 (8 species); native perennial grasses 2 (13 species); and non-native perennial grasses (3 species). In trial II, plots were seeded to monocultural plots of 15 accessions of native Californian and three cultivars of non-native perennial grasses. Plots in both trials were seeded during January 1992 and evaluated for three successive years.

Notes: We studied the utility of gap formation and soil disturbance as methods to enhance establishment of plant species in the understory of a northern Kentucky forest where Lonicera maackii (Amur honeysuckle) produced dense thickets. In May 1994, gaps (5 m diameter) were cut in the shrub thicket. In adjacent areas, the shrub canopy remained intact. Subplots were established where soil was either turned with a spade to a depth of 15 cm or not disturbed. We monitored plant establishment for three growing seasons (1994, 1995, and 1996). Shrub removal increased light availability to about 10% of full sun. Gap formation had a significant (p 〈 0.05) and positive influence on total plant density (exclusive of L. maackii), and soil disturbance did not (p 〉 0.05). After three growing seasons, the most important species were L. maackii, Alliaria petiolata, Parthenocissus quinquefolia, Vitis vulpina, and Acer negundo. Of these species, only V. vulpina showed significantly (p 〈 0.05) higher densities in gaps. Other less important species such as Phytolacca americana, Campsis radicans, and Eupatorium rugosum occurred almost exclusively in gaps. Of the 44 taxa observed in this study, most were generalist species that also occur in early successional habitats. Long-term dominance of the understory by L. maackii has likely modified system attributes with corresponding effects on community development. Shrub removal provides a window of establishment for various plant species, but successful restoration may require further management species availability and to control new invaders.

Notes: Many forest roads are being closed as a step in watershed restoration. Ripping roads with subsoilers or rock rippers is a common practice to increase the infiltration capacity of roads before closure. When considering the effectiveness of ripping for reducing runoff and erosion and the potential reduction in slope stability by saturating road fills, it is important to know how ripping changes the infiltration capacity of forest roads. Hydrographs from simulated rainfall on 1 × 1 m plots were analyzed to find the saturated hydraulic conductivity, an indicator of infiltration capacity. I examined saturated hydraulic conductivity for three treatments on two different soils. One road was built in a soil derived from the metamorphic belt series geology of northern Idaho, a soil noted for its high rock fragment content. The second road was built in a sandy soil derived from decomposed granitics of the Idaho batholith. On each soil, five plots were installed on a road before ripping, and nine plots were installed on the same road segment following ripping, four covered with a heavy straw mulch and five without. Three half-hour rainfall events with intensities near 90 mm/hr were simulated on each plot. Results show that ripping increases hydraulic conductivities enough to reduce risk of runoff but does not restore the natural hydraulic conductivity of a forested slope. The unripped road surfaces had hydraulic conductivities in the range of 0–4 mm/hr, whereas ripped roads were in the range of 20–40 mm/hr after the second event. Surface sealing and tilled soil subsidence processes are important in reducing the hydraulic conductivity of the soils with repeated wetting. Subsidence appears to be important on the granitic soil, whereas surface sealing was more important on the belt series soil.

Notes: Rainforest restoration is a relatively new endeavor, and few attempts have been made to assess the success of such restoration efforts in terms of the reestablishment of an ecosystem. Small plantings of rainforest tree species have been carried out adjacent to mature rainforest at Lake Barrine National Park in North Queensland, Australia, since 1988. The aim of this project was to assess the leaf litter invertebrate fauna of these plantings as indicators of the success of the restoration process. Plots planted in 1988, 1989, and 1990, as well as adjacent mature rainforest, were sampled in the wet and dry seasons of 1994. Invertebrates were extracted from leaf litter samples with Berlese Funnels and sorted to order. Diversity, the abundance of different size classes and orders of invertebrates, and the abundance of different functional groups were examined. In most respects the 1988 plot was found to differ little from the mature rainforest plots, whereas the 1990 plot lacked small and predatory invertebrates, especially in the dry season. The 1989 plot was intermediate in invertebrate abundance and diversity. The use of partially deciduous trees in the 1989 and 1990 plots, resulting in lower canopy cover at the driest time of the year, may have contributed significantly to the differences found between the early and later plantings. It is recommended that trees that provide good canopy cover year-round be used as dominant species in plantings to facilitate the development of a leaf litter ecosystem that can be sustained throughout the year.

Notes: Tidal wetland mesocosms at Tijuana River National Estuarine Research Reserve failed to elucidate effects of hydrologic treatments (excluded, impounded, and fully tidal systems) for most parameters measuring Salicornia virginica (pickleweed). Although soil salinity increased where tidal flushing was excluded for 10 months (salinities rose ∼20 to 50%), pickleweed cover and algal chlorophyll did not differ among treatments. Effects were seen only in pickleweed growth rates (∼30% decrease where tides were excluded) and normalized difference vegetation index (NDVI) measurements. We failed to show any differences between impounded and fully tidal conditions, because the mesocosms had coarse sediments, and impounded water drained easily via subsurface flow. However, the problems that we encountered with the mesocosms led to the following advice for future wetland restoration projects: (1) Mesocosms are useful for testing restoration techniques before an actual restoration project takes place. (2) Mesocosms should be used to test factors that may lead to more successful restoration in the future, including planting techniques, substrate conditions, and hydrology. (3) Mesocosms should be used to develop new assessment methods for monitoring wetland ecosystems. Because of the ability to control some environmental parameters while maintaining seminatural conditions, mesocosms offer great potential for the future evaluation of experimental restoration techniques.

Notes: Habitat protection is a major component of the Exxon Valdez oil spill restoration process. The acquisition of private lands, or partial interests in private lands, is intended to promote natural recovery of spill-injured resources and services by removing the threat of additional development impacts. The Comprehensive Habitat Protection Process is the method that was designed to achieve this objective. Over one million acres within the oil spill affected area were evaluated, scored, and ranked by a multi-criteria evaluation process. Initially, lands were divided into large parcels encompassing entire bays and watersheds. Criteria were then used to assess the habitat and human-use values associated with each parcel and the protection benefit that acquisition would provide for 19 injured resources and services. This process has been the basis for the acquisition of 41,549 acres of land on Afognak Island and 23,800 acres on the Kenai Peninsula and for agreements that, if consummated, will result in the acquisition of fee or lesser rights on over a half million acres of land in the Kodiak Archipelago, on the Kenai Peninsula, and in Prince William Sound. All of these lands or rights, if acquired, will be incorporated into parks or refuges or otherwise managed in a manner that will facilitate the recovery of the resources and services injured by the oil spill.

Notes: Agricultural set-aside and compensation land provide restoration ecologists with opportunities to re-create semi-natural habitats. Restoration sites often have high soil fertility and inadequate seed banks of desirable species. Sowing additional seed is a proven method for establishing chalk grassland vegetation. If seed of local provenance is required, it may be collected by hand or by using specialized machines. Ultimately, the mix collected must provide seed suitable for recreating vegetation similar to that of the donor site. We examine the ability of a vacuum machine to meet this requirement by comparison with hand collection, and we discuss possible effects on invertebrates. Microscope analysis and glasshouse and field trials were used to compare the abundance of seed of different species in harvested mixes with the vegetation composition of the donor site. Seed heads of individual species were examined to determine the number of viable seeds per head and attack rates by phytophagous insects. The mix contained seed of over half the species recorded on the donor site. The seed of taller, more common species was overrepresented in the mix, at the expense of some smaller, mat-forming plants. After one season, however, the vegetation of the field trial plots was of the same type as that of the donor site, although the proportions of the constituent species differed slightly and certain species were absent. Mechanical collection is more efficient than hand collection. Endophagous invertebrates are unlikely to be affected by the machine. Seed collection requires a combination of methods, precise timing, and careful planning to provide a full range of species and to minimize impacts on plant and invertebrate populations.

Notes: Abandoned limestone quarries are hostile environments for plant and invertebrate colonization and establishment. The length of time taken for successful establishment by natural processes may be unacceptable for reclamation purposes; several techniques are used to reduce the time scales involved. A new technique, restoration blasting, aims to replicate natural daleside landforms by selective blasting of modern production quarry faces. We compare the flora and invertebrate fauna of restoration-blasted sites, hydro-seeded with daleside species, with naturally regenerating disused quarries and a natural daleside. Restoration-blasted sites were found to have less plant cover, more bare ground, fewer orders of invertebrates, and generally fewer animals within each order than the other two types of site. The disused quarries tended to have intermediate characteristics between the restoration-blasted sites and the natural daleside. The age of the site may be important in determining the plants and invertebrates occurring there. This may be related to the time available for establishment or a greater degree of settlement or stability within the biotic and abiotic components of the site. Although most of the results indicate that time since establishment may be important, some variations occur. In particular, the development of vegetation cover in areas grazed by rabbits is problematic. These results are important in the assessment of successful reclamation because the invertebrate fauna may contribute greatly to the overall system. Both plant and animal communities appear to be establishing well on the sites reclaimed by restoration blasting. Further monitoring will identify the speed at which such environments achieve the desired aim of replicating daleside communities and the communities best able to be sustained following this technique.

Notes: . Gram- symbiotic X-bacteria present in the xD strain of Amoeba proteus as required cell components, synthesize and export a large amount of a 29-kDa protein (S29x) into the host's cytoplasm across bacterial and symbiosome membranes. The S29x protein produced by E. coli transformed with the s29x gene is also rapidly secreted into the culture medium. Inside amoebae, S29x enters the host's nucleus as detected by confocal and irnmunoelectron microscopy, although it is not clear if S29x is selectively accumulated inside the nucleus. The deduced amino-acid sequence of S29x has a stretch of basic amino acids that could act as a nuclear localization signal, but there is no signal peptide at the N-terminus and the transport of S29x is energy independent. The functions of S29x are not known, but in view of its prominent presence inside the amoeba's nucleus, S29x is suspected to be involved in affecting the expression of amoeba's nuclear gene(s).

Notes: . We used a photoactivatable fluid-phase marker to isolate a new collection of thermosensitive endocytosis mutants in the cellular slime mold Dicfyostelium discoideum. All the strains were thermosensitive for growth on bacteria or axenic medium at 27° C. Initial rates of endocytosis rapidly decreased upon incubation at the restrictive temperature, but surprisingly most of the strains showed a transient recovery of activity with prolonged exposure to 27° C. Endocytosis and exocytosis activities were uncoupled for some of the cell lines at 27° C whereas the others had to be shifted to 29° C. Further molecular analysis of these mutants could lead to the discovery of new proteins involved in endocytosis and its regulation.

Notes: . Monoclonal antibodies against spores of Glugea atherinae were obtained after lymphocytic hybridization made from immunized mouse splenocytes. Screening using an indirect enzyme linked immunosorbent assay (ELISA), revealed seven monoclonal antibodies with an intense but variable reaction with the spores of fish microsporidia, and a moderate reaction with those of an insect microsporidium (Nosema sp.). The reaction was weaker with spores of Encephalitozoon intestinalis found in HIV' patients. FITC and Dot Blot confirmed the majority of these results. After biotinylation of the seven antibodies, inhibition tests allowed the localization of two different recognition domains on the spores of Glugea atherinae. The multiple antigenic determinants and their probable polysaccharide nature seem to be in accord with the class IgM of the antibodies produced. This work confirms the potential of these antibodies for microsporidian taxonomy and diagnosis, especially the use of Mabs 12F9 and 12H5 for detection of spores in stools of HIV+ patients.

Notes: . Leidyana canadensis n. sp. (Apicomplexa: Eugregarinida) is described from larvae of the eastern hemlock looper, Lambdina fiscellaria (Lepidoptera: Geometridae) collected near St. Stephen, Charlotte County, New Brunswick, Canada. Gamonts solitary, located between host ventricular peritrophic membrane and ventricular epithelium. Protomerite very broadly ellipsoid with transverse posterior margin; length 43.7 μm, width 42.7 μm; protomerite-deutomerite septum strongly constricted. Deutomerite narrowly obovoid with anterior transverse margin; length 186.6 μm, width 58.7 μm; total length 227.1 μm. Nucleus spherical to broadly ellipsoid; length 26.3 μm, width 20.2 μm; placement abaxial in the posterior 2/3 of the deutomerite. Nucleus often obscured in late gamonts. Association late, caudofrontal, ephemeral and leading directly to syzygy. Gametocysts roughly spherical; diameter 216.7 μm; hyaline coat increasing diameter to 359.1 μm. Gametocysts mature and dehisce through six short spore tubes within 48 hours. Oocysts axially symmetric, dolioform in dorsal aspect, compressed in the plane perpendicular to the major axis, very uniform in size and shape; length along major axis 5.2 μm, terminal width 1.8 μm, medial width 3.8 μm; extruded in chains.

Notes: . Distribution of membrane cholesterol at the attachment site of Cryptosporidium muris was investigated by freeze-fracture cytochemistry using a polyene antibiotic filipin. Since the host plasma membrane enveloped C. muris, the inner and outer membranes were continuous with the parasite plasma membrane at the annular ring and with host membrane at the dense band, respectively. Although many filipin-cholesterol complexes were observed on the plasma membrane of host cells and parasites, a line showing no complexes was evident at the above two membrane junctures. These observations indicate that parasitic infection of C. muris altered the organization of membrane cholesterol.

Notes: . Cultivation of Pneumocystis carinii from rabbit onto cell monolayers was intended. Three cell types were used: rat lung derived cell line L2 and canine kidney derived cell line MDCK, and primary epithelial rabbit lung cells derived from 20 days old foetuses. These primary cells were also immortalized by transfection of the coding sequences for the large and small T antigens of the SV40 virus. P. carinii were extracted from lungs of corticoid treated young rabbits and were purified on Ficoll. In vitro development of P. carinii was assessed by counting the number of attached parasites on cells after staining. No development was observed on L2 and MDCK cell lines. On the contrary, a development was observed on rabbit derived cells with a threefold increase of attached parasites on the third day of culture. Immortalized cells allowed also multiplication of attached P. carinii. These results are similar to those obtained with culture of rodent P. carinii onto cell monolayers.

Notes: . A large number of developmentally regulated DNA rearrangements occur during the development of the macronucleus in Tetrahymena thermophila, Tlr1 is a deletion element which has large inverted repeats near the rearrangement junctions and deletes more than 13 kbp of internal DNA. Previous analysis of caryonidal lines revealed alternate left junctions for the Tlr1 rearrangement in B strain cells. We show here that C2 strain Tetrahymena also use alternate rearrangement junctions. We have mapped and sequenced two additional rearrangement variants and find that both the left and right can vary over a range of approximately 200 bp. We also demonstrate the presence of sequence microheterogeneity in the most commonly found Tlr1 rearrangement product.

Notes: . In recognizing food particles, Dictyostelium cell-surface molecules initiate cytoskeletal rearrangements that result in phagosome formation. After feeding D. discoideum cells latex beads, early phagosomes were isolated on sucrose step gradietns. Protein analyses of these vesicles showed that they contained glycoproteins and surface-labeled species corresponding to integral plasma membrane proteins. Cytoskeletal proteins also were associated with phagosomes, including myosin II, actin and a 30 kDa-actin bundling protein. As seen by the acridine orange fluorescence of vesicles containing bacteria, phagosomes were acidified rapidly by a vacuolar H+-ATPase that was detected by immunoblotting. Except for the loss of cytoskeletal proteins, few other changes over time were noted in the protein profiles of phagosomes, suggesting that phagosome maturation was incomplete. The indigestibility of the beads possibly inhibited further endocytic processing, which has been observed by others. Since nascent phagosomes contained molecules of both the cytoskeleton and plasma membrane, they will be useful in studies aimed at identifying specific protein associations occurring between membrane proteins and the cytoskeleton during phagocytosis.

Notes: . In situ hybridization using fluorescent oligonucleotide probes complementary to unique regions of 16S rRNA molecules provides a way of identifying the food vacuole contents of bactivorous protists. Laboratory experiments with Tetrahymena showed rRNAs in food vacuoles are degraded slowly enough to permit their use as hybridization targets for such probes. A probe specific for a hypervariable region of the small subunit rRNA of an unnamed proteobacterium abundant in a local lake was then synthesized. It was used to probe the food vacuoles of the ciliates present in fixed water samples collected from the same lake. The vacuoles of several filter-feeding ciliates bound the probe, indicating that such probes can be used to identify the food vacuole contents of ciliates collected from natural samples.

Notes: . Oxymonas has the characteristic structures and organization of other oxymonads including two separated pairs of basal bodies/flagella, a preaxostylar lamina, a paracrystalline axostyle, and an absence of mitochondria and Golgi. Like other Oxymonadinae genera it possesses a long proboscis, the rostellum which is terminated by the holdfast. Like the genera Pyrsonympha and Streblomastix, Oxymonas possesses a holdfast which permits it to attach to the cuticle of the termite hind-gut. This holdfast is subdivided into rhizoids and is filled with microfilaments. The rostellum is variable in length and contains two distinct microtubular bundles. One bundle is composed of convoluted microtubular ribbons which originate at the base of the holdfast and extend posteriorly along the rostellum and before penetrating into the cell body. The second bundle is composed of flexuous free microtubules which originate at different levels of the rostellum, increasing in number from top to base. They occupy the axial part of the rostellum and incorporate into the axostylar rows at the basal body/flagellar level. Microtubules of the paracrystalline axostyle are cross-linked by bridges forming parallel rows like in the contractile axostyles of other oxymonads such as Pyrsonympha and Saccinobaculus. Most of the microtubules of the axostyle originate at the flagellar/preaxostylar level but some originate from the axial flexous free microtubules of the rostellum, as indicated above. The possibility of an extension/retraction of the rostellum, suggested by other authors, is discussed.

Notes: . The effects of platelet-activating factor (PAF), at doses ranging from 10−6 M to 10−10 M, on cell growth and on cell differentiation of Herpetomonas muscarum muscarum were investigated. Cell differentiation was evaluated by both light and electron microscopy. At the concentrations used, PAF did not interfere with the protozoan growth. However, parasites grown in the presence of PAF (10−6 M) were significantly more differentiated than those grown in the absence of PAF, since the first day of culture. On the first two days of culture, PAF doses ranging from 10−10 M to 10−7 M, did not significantly interfere with the differentiation of these parasites, although after the third day of culture, all PAF doses used significantly increased the protozoan differentiation. Specific PAF receptor antagonists totally abrogated (WEB 2086 and WEB 2170)or significantly decreased (BN 52021) PAF effect on cell differentiation. These findings indicate PAF triggers the process of cell differentiation in Herpetomonas muscarum muscarum and suggest these parasites have receptors for PAF.

Notes: . In this study we have examined the distribution of epitopes defined by monoclonal antibodies raised against Trypanosoma cruzi amastigotes during the intraceullar life cycle of the parasite. We have raised monoclonal antibodies towards amastigote forms and performed preliminary immunochemical characterization of their reactivities. MAB 1D9, 3G8, 2B7, 3B9, and 4B9, and 4B9 react with carbohydrate epitopes of the parasite major surface glycoprotein—Ssp-4 defined by MAB 2C2 [5]: MAB 4B5 reacts with a noncarbohydrate epitope in all developmental stages of the parasite, and MAB 3B2 also detects a noncarbohydrate epitope preferentially in T. cruzi flagellared forms. Vero cells infected with tissue culture-derived trypomastigotes of clone D11 (G strain) were fixed at different times during the intraceullular proliferation of parasites, and processed for immjno-electron microscopy and confocal immunoflurescence with the different monoclonal antibodies. We observed that while the surface distribution of MAB 2C2 and 4B9 epitopes was uniform throughout the cycle, MAB 1D9, 3G8, and 2B7 reacted with cytoplasmic membrance-bound compartments of the amastigotes. MAB 3B9 displayed a unique surface dentate pattern in some amastigotes. MAB 4B5 recognized a curved-shaped structure at the flagellar pocket region in some intracellular amastigotes and localized to the membrane in dividing forms. In intracellular trypomastigotes, MAB 4B5 also displayed a punctate pattern near the flagellar pocket.

Notes: . The fate of the [methyl-14C] group of S-adenosylmethionine (AdoMet) in bloodstream forms of Trypanosoma brucei brucei, was studied. Trypanosomes were incubated with either [methyl-14C]methionine, [U-14C]methionine, S-[methyl-14C]AdoMet or [35S]methionine and incorporation into the total TCA precipitable fractions was followed. Incorporation of label into protein through methylation was estimated by comparing molar incorporation of [methyl-14C] and [U-14C]methionine to [35S]methionine. After 4-h incubation with [U-14C]methionine, [methyl-14C]methionine or [35S]methionine, cells incorporated label at mean rates of 2,880 pmol, 1,305 pmol and 296 pmol per mg total cellular protein, respectively. Cells incubated with [U-14C] or [methyl-14C]methionine in the presence of cycloheximide (50 μg/ml) for four hours incorporated label eight- and twofold more rapidly, respectively, than cells incubated with [35S]methionine and cycloheximide. [Methyl-14C] and [U-14C]methionine incorporation were 〉 85% decreased by co-incubation with unlabeled AdoMet (1 mM). The level of protein methylation remaining after 4-h treatment with cycloheximide was also inhibited with unlabeled AdoMet. The acid precipitable label from [U-14C]methionine incorporation was not appreciably hydrolyzed by DNAse or RNAse treatment but was 95% solubilized by proteinase K. [U-14C]methionine incorporated into the TCA precipitable fraction was susceptible to alkaline borate treatment, indicating that much of this label (55%) was incorporated as carboxymethyl groups. The rate of total lipid methylation was found to be 1.5 times that of protein methylation by incubating cells with [U-14C]methionine for six hours and differential extraction of the TCA lysate. These studies show T. b. brucei maintains rapid lipid and protein methylation, confirming previous studies demonstrating rapid conversion of methionine to AdoMet and subsequent production of post-methylation products of AdoMet in African trypanosomes.

Notes: . A species of peritrich that attaches to gills of the skate, Raja erinacea, was identified by its original describer as a member of Caliperia, a genus characterized by having a noncontractile skeletal rod within the arms of its cinctum and by not having the cinctal arms bonded to one another at their tips. Our observations of the living ciliates confirmed by protargol impregnation and electron microscopy revealed that their cinctal arms are linked by a bouton and that the cytoskeletal structure within them has the fine structure of a myoneme. These characteristics place this peritrich unequivocally in the genus Ellobiophrya and it is thus renamed Ellobiophrya brevipes (Laird, 1959) n. comb. Clumps of epithelial cells clasped by the cincta of E. brevipes show damage at their bases but not on their luminal surfaces. The known species of Ellobiophrya are compared for significant structural differences that separate species of this genus.

Notes: . The composition and incorporation of lipids in two marine ciliates, Pleuronema sp. and Fabrea salina, was examined following growth on either an algal or bacterial diet. When allowed to feed on a natural bacterial community, Pleuronema sp. synthesized the triterpenoid alcohol gammaceran-3β-ol (tetrahymanol) and two hopanoids (hopan-3β-ol and one uncharacterized hopanoid). When fed the marine alga Isochrysis galbana, F. salina contained the major algal sterol 24-methylcholesta-5, 22-dien-3β-ol and several long chain ketones specific to the alga. In both ciliates, fatty acids composition showed a general correspondence to that of the diet. Using a series of antibiotic treatments to alter the bacterial prey community, and thus fatty acid composition of the ciliate's diet, promoted changes in the fatty acid composition of Pleuronema sp. to resemble that of the bacterial prey. The addition of a mixture of algal sterols to a bacterized culture of another scuticociliate, Parauronema acutum, inhibited tetrahymanol synthesis and resulted in the incorporation of sterols into the ciliate.

Notes: . Ciliates are often important members of aquatic communities in terms of their biomass, productivity, trophic roles, or numerical abundance. The interaction of metazoan predators with ciliates will be mediated by a number of biotic factors, including the potential of ciliate populations for growth, the relative size of ciliates and metazooplankton, the species structure of the metazooplankton, and the defenses of ciliates. This paper reviews some of the recent laboratory an field data pertaining to these particular factor. Studies have generally shown that metazoans can reduce ciliate population growth rates, but this impact varies greatly with the ciliate and metazoans involved. Smaller ciliates are generally more vulnerable to metazoan predators than larger species, although this relationship will be affected by the defenses a ciliate may possess. The structure of the metazooplankton community itself will also affect ciliatemetazoan interactions. The suppression of ciliate populations by metazoans has important ecological consequences, and more study is needed to understand the interaction of these groups in aquatic systems.

Notes: . Survival of a potential prey organism depends on the effectiveness of its physical, chemical, behavioral and life history responses to the appearance of a predator. Inducible defenses are flexible responses in which predator (or competitor)-released substances stimulate potential prey organisms to transform into predator-resistant phenotypes. Induced defenses may be highly protective. Benefits however are often balanced by fitness costs such as decreased growth rates or reduced reproductive potential. Here I discuss inducible defenses in ciliates with particular attention to the hypotrich genera: Aspidisca, Euplotes, Onychodromus, Sterkiella, and an undescribed hypotrich genus. I isolated Sterkiella sp. and the undescribed genus from vernal woodland pools on Saint Anselm College campus. Experimental evidence shows that a signal-induced defensive transformation occurs in these ciliates within hours after exposure to a predator cue and results in a significant decrease in susceptibility to predation. Deployment of ciliate antipredator structures such as spines, keels, ridges and other protuberances requires a large investment of cytoskeletal elements, primarily microtubules, and incurs an evolutionary cost in the form of significantly reduced growth rates. Onychodromus quadricornutus exhibits an extraordinary degree of phenotypic plasticity. In response to different environmental conditions individuals within a clone may express one of three general phenotypes: basic, lanceolate, or giant cells. The predacious giant phenotype releases a morphogenetically active signal substance, Onychodromus-factor, that triggers defensive phenotypic transformation in both intraspecific and interspecific prey. Enzyme degradation and ultrafiltration experiments indicate that Onychodromus-factor is a peptide with a molecular weight below 10,000 Da. Conspecifics develop hypertrophied dorsal spines when exposed to Onychodromus-factor. Sterkiella cells develop two defensive dorsal keels and transform to an enlarged ovoid cell in response to Onychodromus-factor as well as inducing signals released by Stylonychia, Urosyla, and Lembadion. Field studies of two vernal pools show that defensive phenotypic transformation in Sterkiella cells coincides with the appearance of Lembadion magnum during vernal pool succession. An undescribed hypotrich genus also expresses its defended phenotype when Lembadion is present in these pools. Aspidisca turrita (Ehrenberg, 1838) Claparede and Lachmann 1858, closely resembles Aspidisca lynceus (Müller, 1773) except for the possession of a dorsal thorn-like structure. Experimental evidence shows that the dorsal thron is a defensive structure induced by signals released by the predacious ciliates Urostyla grandis and Lembadion magnum. Thus, A. turrita and A. lynceus are alternate phenotypes of the same species. I speculate that inducing signals function in predacious ciliates as lectin-like, carbohydrate-binding adhesion proteins during prey recognition and that prey species have evolved specialized cell surface receptors that allow detection of different predator proteins. I consider consequences for both predator and prey.

Notes: . The development of Toxoplasma gondii was studied in mice fed bradyzoites. At one hour after oral inoculation (HAI), bradyzoites were found in cells of the surface epithelium and the lamina propria of the small intestine, primarily the ileum. Division into two tachyzoites was first observed at 18 HA1 in the intestine. At 24 HAI, organisms were also seen in mesenteric lymph nodes. Organisms were first detected in the brain at six days after oral inoculation with bradyzoites (DAI) but not consistently until 10 DAI. Immunohistochemical staining with bradyzoite specific (BAG-5 antigen) anti-serum showed that bradyzoites retained their BAG-5 reactivity even after the first division into two tachyzoites in the intestine at 18 HAL BAG-5 positive organisms were not seen 2–5 DAI. BAG-5 antigens reappeared in T. gondii at 6 DAI. Whole mice and individual tissues of mice fed bradyzoites were bioassayed in cats and mice for the presence of bradyzoites. Feces of cats fed murine tissues were examined for oocyst shedding for short prepatent periods. Bradyzoites were present in the intestines of mice up to 12 HA1 but not at 18 HAI, and tachyzoites and not bradyzoites disseminated to other tissues from the intestine. Bradyzoites were again detected 6 DAI. Using the mouse bioassay, T. gondii was first detected in peripheral blood at 24 HA1 and more consistently at 48 HAL Using a pepsin-digestion procedure and mouse bioassay, organisms were demonstrated in many tissues of mice 15 and 49 DAI.

Notes: . The release of soluble acid phosphatase from Leishmania donovani promastigotes in axenic culture medium is altered by osmolarity changes: Hypoosmotic conditions increase its release whereas hyperosmotic ones decrease the release of acid phosphatase.

Notes: . The alternating myxosporean and actinosporean stages of the myxozoan parasitc Myxobolus cerebralis (Hofer 1903) from its salmonid fish and aquatic oligochaete hosts, respectively, were compared for sequence homology of the small subunit (18S) ribosomal RNA genes. A 99.8% similarity between the sequences of these two stages was substantially greater than that of M. cerebralis compared to two other Myxobolus sp. from salmonid fish. Our results are the first molecular evidence confirming the alternating stages initially described by Wolf and Markiw [25] for the life cycle of M. cerebralis but found in two different taxonomic classes (Myxosporea and Actinosporea) are indeed forms of the same organism. Sequencing of rRNA genes of the actinosporean stage followed by development of specific primers for DNA amplification of the myxosporean stage, as in our study, should be applied to solve other myxozoan life cycles. Additionally, these approaches will in the future provide useful diagnostic reagents for the detection and study of this important group of fish pathogens.

Notes: . The secretion of cellulose-degrading enzymes by Leishmania promastigotes in culture and in the sandfly vector was demonstrated. Two types of activity of cellulase enzyme-complexes were measured: endoglucanases, which randomly cleave cellulose chains and cellobioydrolases, which remove cellobiose from the nonreducing end of the molecule. The assays demonstrated that enzymes with these activities were secreted into the culture medium by Leishmania major, L. donovani, and L. braziliensis. These activities were also found in cultures of Sauroleishmania agamae, Leptomonas seymouri, Herpetomonas muscarum, Crithidia fasciculata and Trypanosoma brucei brucei that had a relatively low endoglucanase activity. Both endoglucanase and cellobiohydrolase activities were found in the gut of L. major-infected Phlebotomus papatasi, while gut preparations of uninfected sandflies had only cellobiohydrolase activity. The similar growth of L. major parasites in medium supplemented with either cellulose or glucose suggests these parasites can utilize cellulose.

Notes: . Sequences of the small subunit rRNA genes of Amblyospora californica and an Amblyospora sp. from Culex salinarius were determined. These sequences were compared phylogenetically with 16 other microsporidia. The results suggest Amblyospora forms a sister taxon to the rest of the microsporidia examined. The basal position of Amblyospora is discussed with respect to the evolution of microsporidian life cycles. These sequences represent the longest microsporidian small subunit rRNA genes sequenced to date, 1,359 and 1,358 bp, respectively. Structural features and GC content (49% for both) are comparable to those of other microsporidia which have been sequenced.

Notes: . The cell surface of the synhymeniid ciliate, Zosterodasys agamalievi, consists of shallow kinetal grooves separated by low cortical ridges. Numerous electron-opaque bodies are located in the cortical ridges, inside the kinetal grooves, and are distributed in parallel rows between adjacent kineties. Well-developed alveoli are present beneath the cell surface membrane. Zosterodasys agamalievi has a single micronucleus and a homomerous macronucleus. The infraciliature of the somatic monokinetid consists of an anteriorly-directed kinetodesmal fiber, a well-developed divergent postciliary microtubular ribbon, radially-oriented transverse microtubules, and a short striated rootlet, which extends anteriorly from the base of the kinetosome into the cell. Zosterodasys agamalievi has a perioral band of paired cilia, the synhymenium, that winds obliquely across the ventral surface of the body, just posterior to the cytostome. The infraciliature of the anterior kinetosome of the synhymenium consists of two postciliary microtubules; a well-developed, divergent post-ciliary ribbon of microtubules and a short kinetodesmal fiber are associated with the posterior kinetosome. The cytopharynx is supported by 14-16 nematodesmata which are capped distally by a capitulum. The cytopharynx is bound proximally by a fibrous sheath and is lined by radially-arranged microtubular ribbons. No obvious oral ciliature is present.

Notes: . The presence of 14 enzymes was investigated using purified spores of the microsporidian Nosema grylli from fat body of the crickets Gryllus bimaculatus. Glucose 6-phosphate dehydrogenase (EC 1.1.1.49), phosphoglucomutase (EC 5.4.2.2), phosphoglucose isomerase (EC 5.3.1.9), fructose 6-phosphate kinase (EC 2.7.1.11), aldolase (EC 4.1.2.13), 3-phosophoglycerate kinase (EC 2.7.2.3), pyruvate kinase (EC 2.7.1.40) and glycerol 3-phosphate dehydrogenase (EC 1.1.1.8) were detected with activities of 15 ± 1, 7 ± 1, 1,549 ± 255, 10 ± 1, 5 ± 1, 16 ± 4, 6 ± 1 and 16 ± 2 nmol/min. mg protein, respectively. Hexokinase (EC 2.7.1.1), NAD-dependent malate dehydrogenase (EC 1.1.1.37), malic enzyme (EC 1.1.1.40), lactate dehydrogenase (EC 1.1.1.27), alcohol dehydrogenase (EC 1.1.1.1) and succinate dehydrogenase (EC 1.3.99.1) were not detectable. These results suggest the catabolism of carbohydrates in microsporidia occurs via the Embden-Meyerhof pathway. Glycerol 3-phosphate dehydrogenase may reoxidize NADH which is produced by glyceraldehyde 3-phosphate dehydrogenase in glycolysis.

Notes: . We have determined the DNA sequence of the gene encoding the protein of the plasma membrane Ca2+-ATPase in Paramecium tetraurelia. The predicted amino acid sequence of the plasma membrane Ca2+-ATPase shows homology to conserved regions of known plasma membrane Ca2+-ATPases and contains the known binding sites for ATP (FITC), acylphosphate formation, and calmodulin, as well as the “hinge” region: all characteristics common to plasma membrane Ca2+-ATPases. The deduced molecular weight for this sequence is 131 kDa. The elucidation of this gene will assist in the studies of the mechanisms by which this excitable cell removes calcium entering through voltage gated calcium channels and the pump functions in chemosensory signal transduction.

Notes: . Chemosensory adaptation is seen in Tetrahymena thermophila following prolonged exposure (ten minutes) to micromolar concentrations of the chemorepellents lysozyme or guanosine triphosphate (GTP). Since these cells initially show repeated backward swimming episodes (avoidance reactions) in these repellents, behavioral adaptation is seen as a decrease in this repellent-induced behavior. The time course of this behavioral adaptation is paralleled by decreases in the extents of surface binding of either [32P]GTP or [3H]lysozyme in vivo. Scatchard plot analyses of repellent binding in adapted cells suggests the behavioral adaptation is due to a dramatic decrease in the number of surface binding sites, as represented by decreased Bmax values. The estimated KD values for nonadapted cells are 6.6 μM and 8.4 μM for lysozyme and GTP binding, respectively. Behavioral adaptation and decreased surface receptor binding are specific for each repellent. The GTP adapted cells (20 μM for ten minutes) still respond behaviorally to 50 μM lysozyme and bind [3H]lysozyme normally. Lysozyme adapted cells (50 μM for ten minutes) still bind [32P]GTP and respond behaviorally to GTP. All the behavioral and binding changes seen are also reversible (deadaptation). Neomycin was shown to be a competitive inhibitor of [3H]lysozyme binding and lysozyme-induced avoidance reactions, but it had no effect on either [32P]GTP binding or GTP-induced or avoidance reactions. These results are consistent with the hypothesis that there are two separate repellent receptors, one for GTP and the other for lysozyme, that are independently downregulated during adaptation to cause specific receptor desensitization and consequent behavioral adaptation.

Notes: . Cell division in higher eukaryotes is mainly controlled by CDK (serine/threonine) protein kinases and by other components of these kinase complexes. Previously, we showed homologous kinases also occur in the ciate Paramecium tetraurelia. In this report, a polyclonal antibody was produced against a GST fusion protein which contained the N-terminal third of the previously isolated CDC2FTA protein. It recognized a 36 kDa polypeptide on western blots and did not cross-react with the related 34 kDa polypeptide. This 36 kDa polypeptide showed no affinity for yeast P13suc1 protein. The CDC2PTA level was invariant throughout the cell cycle and decreased slightly when cells entered stationary phase. Indirect immunofluorescence showed CDC2PTA localized in the macronucleus, but not in micronuclei. Native CDC2FTA was immunoprecipitated and the kinase activity was assayed using histone HI as substrate in elutriation synchronized samples. The kinase activity was high during the early stages of the cell cycle and reached a peak at 2.5 h after elutriation, which corresponds to the time of initiation of macronuclear DNA synthesis. This suggests CDC2PTA kinase may be associated with the regulation of macronuclear DNA synthesis. These results allow us to draw concrete comarisons for the first time between the role of CDK in ciliates and higher eukaryotes.

Notes: . Programmed DNA rearrangements, including DNA degradation, characterize the development of the soma from the germline in a number of developmental systems. Pddl p (programmed DNA degradation 1 protein), a development-specific polypeptide in Tetrahymena, is enriched in developing macronuclei (anlagen) and has been implicated in DNA elimination and nucleolar biogenesis. Here, immunocytochemistry and fluorescent in situ hybridization (FISH) were employed to follow Pddl p and two nucleolar markers (Nopp52 and rDNA) during macronuclear development. Both Pdd 1p and Nopp52 localize to subnuclear structures, each of which resemble nucleoli. However, while true nucleoli form and persist during development, Pdd 1p-positive structures are only present for a brief period of macronuclear differentiation. Accordingly, two distinct organelles can be recognized in anlagen: (1) Pdd 1p-positive structures, which lack Nopp52 and rDNA, and (2) developing nucleoli which contain rDNA and Nopp52 but lack Pdd 1p. Taken together with recent data corroborating Pdd 1p's role in DNA elimination, we favor the hypothesis that Pdd 1p structures are unique, short-lived organelles, likely to function in programmed DNA degradation and not in nucleolar biogenesis.

Notes: . The protist Prorocentrum lima, a primary producer of the tumour promoter okadaic acid, is a member of the dinoflagellate class of marine microorganisms. Herein, we have identified and characterized a protein tyrosine kinase (designated PLIK 1A) in P. lima that autophosphorylates almost exclusively on tyrosine residues. PLIK 1A was shown to have an approximate molecular mass of 38 kDa by SDS-PAGE and a native molecular mass within the range of 47–55 kDa by Superdex-75 gel filtration. Phosphoamino acid analysis of autophosphorylated PLIK 1A revealed the presence of phosphotyrosine and autophosphorylated PLJK 1A reacted with monoclonal anti-phosphotyrosine antibodies in a Western immunoblot. In addition, two protein tyrosine phosphatases were identified in P. lima that had apparent molecular masses within the ranges of 150–168 kDa and 73–82 kDa as determined by Superdex-200 gel filtration. These P. lima phosphatases, termed PLPTP-I and PLPTP-II, efficiently dephosphorylated tyrosine phosphorylated myelin basic protein. owever, only PLPTP-I was capable of dephosphorylating the tyrosine phosphorylated substrate angiotensin. Both PLPTP-I and PLPTP-II were able to dephosphorylate tyrosine autophosphorylated PLIK 1A. These data provide the first evidence for reversible tyrosine protein phosphorylation in P. lima by protein tyrosine kinases and phosphatases

Notes: . The energy metabolism was compared among four different representatives of the genus Phytomonas isolated from different plants and localities: the sieve tubes of the hartrot-infected coconut palm in French Guyana, the latex fluid of Euphorbia hyssopifolia in French Guyana and the fruits of tomato and cherimoya in Spain. All four isolates produced acetate, ethanol, glycerol and glycine as metabolic end-products. In addition, small amount of succinate and pyruvate were excreted. Only minor quantitative differences were observed in the four isolates. Glycosomes, harboring the glycolytic enzymes, were present in all isolates. No evidence was found for an active involvement of the mitochondrion in metabolism. Respiration was insensitive to the classical inhibitors of the respiratory chain, such as antimycin and potassium cyanide, but inhibited by salicylhydroxamic acid. No evidence was found for the functioning of a citric-acid cycle. It is concluded that representative of this genus share the same highly active carbohydrate metabolism combined with a complete suppression of mitochondrial activity.

Notes: To establish an in vitro culture system for the precystic phase of Sarcocystis singaporensis, we initially tested various excysting fluids for sporocysts. An excysting fluid containing 2.5% bovine taurocholate and 10% bile of the specific intermediate host, Rattus norvegicus, in RPMI medium was the most suitable resulting in excystation of 80% of the sporozoites. Subsequently, we identified brain endothelial cells and pneumonocytes of the rat to promote growth of sporozoites to schizonts. Hepatoma, fibroblastic, or myoblastic cells were not suitable for the parasite's development. First-generation schizonts were seen at days 3-10 postinoculation (PI); a distinct second peak of schizogonic development only occurred in endothelial cells at days 14-18 PI. First-generation schizonts were 26.0 (± 3.8) μm in diameter and contained 32-50 merozoites, second-generation schizonts measured 34.4 (± 10.6) μm and contained 54-72 merozoites. Merozoite yield at large-scale culture conditions (75 cm2 flasks) using pneumonocytes as host cells was relatively low. Ultrastructurally, sporozoites and merozoites were quite similar to corresponding stages of other Sarcocystis species. With regard to host cell specificity and developmental kinetics, in vitro cultivation showed close similarities to the situation in vivo.

Notes: . The establishment of chloroplasts as cellular organelles in the dinoflagellate, heterokont (stramenopile), haptophyte, and cryptophyte algae is widely accepted to have been the result of secondary endosymbiotic events, that is, the uptake of a photosynthetic eukaryote by a phagotrophic eukaryote. However, the circumstances that promote such associations between two phylogenetically distinct organisms and result in the integration of their genomes to form a single functional photosynthetic cell is unclear. The dinoflagellates Peridinium foliaceum and Peridinium balticum are unusual in that each contains a membrance-bound eukaryotic heterokont endosymbiont. These symbioses have been interpreted, through data derived from ultrastructural and biochemical investigations, to represent an intermediate stage of secondary endosymbiotic chloroplast acquistion. In this study we have examined the phylogenetic origin of the P. foliaceum and P. Balticum heterokont endosymbionts through analaysis of their nuclear small subunit ribosomal RNA genes. Our analyses clearly demonstrate both endosymbionts are pennate diatoms belonging to the family Bacillariaceae. Since members of the Bacillariaceae are usually benthic, living on shallow marine sediments, the manner in which establishment of a symbiosis between a planktonic flagellated dinoflagellate and a botton-dwelling diatom is discussed. In particular, specific environmentally associated life strategy stages of the host and symbiont, coupled with diatom food preferences by the dinoflagellate, may have been vital to the formation of this association.

Notes: . Sexual reproduction of Ascogregarina taiwanensis occurred in pupal Malpighian tubules of its natural host Aedes albopictus, resulting in the formation of gametocysts within which oocysts developed. Sporogony proceeded in each newly formed unsporulated oocyst; eight sporozoites were formed after completion of nuclear divisions followed by the cytokinesis. Developing oocysts were separated by gradient centrifugation on percoll based on different buoyant densities. The slender sporozoite had a typical apical complex composed of a coiled conoid, polar rings, rhoptries with ductules, subpellicular microtubules and micronemes. An apical cavity was seen in the gland-like rhoptries. Mitochondria of gregarines were not seen in any stage during the sporogony. Howeever, amylopectin granules were frequently seen in the cytoplasm. These starch-related granules became scant when the sporozoite was formed. We assumed they were associated with the energy source. Since the apical complex was only present in the sporozoite stage, it was most likely related to the invasion of host epithelial cells of the midgut during the early phase of infection.

Notes: . Surface plaque matrix (PQM) and a tubular arrangement of filaments border Trachipleistophora hominis parasites during growth within host muscle. The PQM at the parasite surface forms a network of processes which can be associated with filamentous tubules. Peroxidase tracer delineated the PQM and showed apparent connections with the tubules. The tubules at the interface of T. hominis-infected cells are structurally similar to the extrasporular tubules of the microsporidian, Ameson michaelis. The extrasporular tubules of A. michaelis and the proteins from T. hominis-infected muscle reacted to keratin antibodies, K8.13, K4 and K13. Conversely, antibodies produced to T. hominis-infected muscle, reacted with the extrasporular tubular proteins of A. michaelis. The PQM and tubular elements are thought to play an important role in affecting molecular traffic between the host and parasite.

Notes: . The fate of methionine in eukaryotic cells is divided between protein synthesis and the branched pathway encompassing polyamine synthesis, methylation of proteins and lipids, and transsulphuration reactions. Aside from protein synthesis, the first step to all other uses of methionine is conversion to S-adenosylmethionine. Blockade of polyamine synthesis in African trypanosomes by the ornithine decarboxylase inhibitor DL-α-difluoromethylomithine (Ornidyl, DFMO) the AdoMet decarboxylase inhibitor 5′-{[(Z)-4-amino-2-butenyl]-methylamino}-5′-deoxyadenosine or the protein methylase inhibitor sinefungin induces dramatic increases in intracellular AdoMet. In a previous study, distribution and pool sizes of [15S] or [U-14C]methionine were followed in bloodform trypanosomes as incorporation into the total TCA precipitable fraction. In the present study, the effects of pretreatment with DFMO (1 mM), MDL 73811 (1 μM) and sinefugin (2 nM) on [35S] and [U-14C]methionine incorporation were studied in blood forms. DFMO or MDL 73811 pretreatment increased protein methylation 1.5-fold through incorporation of [U14C]methionine, while sinefungin caused a 40% reduction of incorporation. The increases in incorporation of [U-14C]methionine due to DFMO and MDL 73811 were reduced 40% to 70% by including cold AdoMet (1 mM) in the incubation medium, an indication of AdoMet transport by bloodform trypanosomes and the utilization of [U-14C]methionine as AdoMet. Exogenous AdoMet had no effect on [35S]methionine incorporation. The agents studied are curative for African trypnosomiasis infections, either clinically (DFMO) or in model infections (MDL 73811, sinefungin) and thus highlight interference with AdoMet metabolism and methylation reactions as biochemical consequences of these agents.

Notes: . Intracellular crystals are conspicuous refractile “inclusion bodies” commonly found in many protozoans, but very few have been identified mineralogically. We have isolated crystals from axenically grown mass cultures of Paramecium tetraurelia, and purified them using differential centrifugation. The crystals’ structure and chemistry were analyzed using x-ray powder diffraction and energy-dispersive electron microprobe techniques. The morphology was studied by means of scanning electron microscopy. The crystals were identified as the orthorhombic mineral, calcian struvite, (Mg, Ca)NH4PO4.6H2O. Struvite from P. tetraurelia exhibited a variety of crystal habits, including hemimorphic forms, epitactic overgrowths and several types of twins. A linear correlation between computed hydration number and Mg content suggests that the crystal composition may reflect the range of conditions under which struvite nucleation and growth occur. The mineral struvite also occurs in association with guano and other rich organic products, and can be biologically induced to precipitate extracellularly. Extracellular struvite has been well characterized in pathogenic calculi (kidney stones) of humans and cats, where precipitation is enhanced by bacterial urease activity that produces ammonia in the urinary tract. This is the first study demonstrating that struvite is also biologically controlled to form as an intracellular mineral. These crystals may have formed within lipid-rich, membrane-bound vesicles in Paramecium.

Notes: . Rules of zoological nomenclature, morphological data, and robosomal DNA sequence data support the validity of the genus Nucleospora, and its placement in the family Enterocytozoonidae. Although Nucleospora exhibits most of the distinguishing morophological characteristics of the family Enterocytozoonidae Cali and Owne, 1990, the distinctively different hosts (fish and humans, respectively) and sites of development (the nuclei of immature blood cells and the cytoplasm of enterocutes) supprot the placement Nucleospora and Enterocytozoon into separate genera. Ribosomal DNA sequence comparisons between Nucleospora salmonis and Enterocytozoon bieneusi showed 19.8% genetic divergece in the large and small subunit regions. Although more inter- and intrageneric divergence between the two species is sufficiently larg to deter suppression of the genus Nucleospora as a junior synonym of Enterocytozoon. A polymerase chain reaction test for the detection of N. salmonis in chinook salmon (Oncorhynchus tshawytscha), based on N. salmonis-specific ribosomal DNA sequence, is described.

Notes: . Lactoferrin and its derived N-terminal peptide may be important host defenses against Giardia lamblia. We showed earlier that lactoferrin and the derived peptides have potent giardicidal activity in vitro. Using indirect immunofluorescence, we now demonstrate binding of lactoferrin and the peptides to the live trophozoite surface. Iron strongly inhibited binding of lactoferrin, and decreased binding of the peptides, while certain divalent metal ions decreased binding of all forms by about half. Lactoferrin and the peptedes caused striking and complex morphologic changes in the trophozoite plasmalemma, endomembranes and cytoskeleton, and increases the electron density of the lysosome-like peripheral vacuoles.

Notes: . Following mating or conjugation, the hypotrichous ciliate Euplotes crassus undergoes a massive genome reorganization process. While the nature of the rearrangement events has been well studied, little is known concerning proteins that carry out such processes. As a means of identifying such proteins, differential screening of a developmental cDNA library, as well as construction of a cDNA subtraction library, was used to isolate genes expressed only during sexual reproduction. Five different conjugation-specific genes have been identified that are maximally expressed early in conjugation, during the period of micronuclear meiosis, which is just prior to macronuclear development and the DNA rearrangement process. All five genes are retained in the mature macronucleus. Micronuclear, macronuclear, and cDNA clones of one gene (conZ47) have been sequenced, and the results indicate that the gene encodes a putative DNA binding protein. In addition, the presence of an internal eliminated sequence in the micronuclear copy of the conZ47 gene indicates that this conjugation-specific gene is transcribed from the old macronucleus.