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: One third of a collection of cloned Stylonychia pustulata micronuclear DNA PstI fragments were found to be of a similar size, consistent with their being members of a repetitious sequence family with a repeat size of about 160 base pairs. Cross-hybridization experiments confirmed that these small cloned fragments are related by sequence homology. Hybridization of the cloned repetitious sequences to PstI digested micronuclear DNA revealed a “ladder” of bands (step size = 160 base pairs), indicating that the repeats are found in tandem arrays. This is the first demonstration of highly repetitious, tandemly repeated sequences in a ciliated protozoan.

Notes: The erythrocytic developmental cycle of Plasmodium falciparum can be conveniently divided into the ring, trophozoite, and schizont stages based on morphology and metabolism. Using highly synchronous cultures of P. falciparum, considerable variation was demonstrated among these stages in sensitivity to chloroquine. The effects of timed, sequential exposure to several clinically relevant concentrations of chloroquine were monitored by three techniques: morphological analysis, changes in the rate of glucose consumption, and changes in the incorporation of 3H-hypoxanthine into parasite nucleic acids. All three techniques gave essentially identical results. The trophozoite and schizont stages were considerably more sensitive to the drug than ring-stage parasites. Chloroquine sensitivity decreased as nuclear division neared completion. The increase in chloroquine sensitivity was coincident with a marked rise in the rate of glucose consumption and nucleic acid synthesis. The rate of nucleic acid synthesis decreased as schizogony progressed while glucose consumption continued at high rates during this process. The degree of chloroquine sensitivity was not highly correlated with either metabolic activity.

Notes: Bloodstream trypomastigote and culture procyclic (insect midgut) forms of a cloned T. rhodesiense variant (WRATat 1) were tested for agglutination with the lectins concanavalin A (Con A), phytohemagglutinin P (PP), soybean agglutinin (SBA), fucose binding protein (FBP), wheat germ agglutinin (WGA), and castor bean lectin (RCA). Fluorescence-microscopic localization of lectin binding to both formalin-fixed trypomastigotes and red cells was determined with fluorescein isothiocyanate (FITC)-conjugated Con A, SBA, FBP, WGA, RCA, PNA (peanut agglutinin), DBA (Dolichos bifloris), and UEA (Ulex europaeus) lectins. Electron microscopic localization of lectin binding sites on bloodstream trypomastigotes was accomplished by the Con A-horseradish peroxidase-diaminobenzidine (HRP-DAB) technique, and by a Con A-biotin/avidin-ferritin method. Trypomastigotes, isolated by centrifugation or filtration through DEAE-cellulose or thawed after cryopreservation, were agglutinated by the lectins Con A and PP with agglutination strength scored as Con A 〈 PP. No agglutination was observed in control preparations or with the lectins WGA, FBA or SBA. Red cells were agglutinated by all the lectins tested. Formalin-fixed bloodstream trypomastigotes bound FITC-Con A and FITC-RCA but not FITC-WGA, -SBA, -PNA, -UEA or -DBA lectins. All FITC-labeled lectins bound to red cells. Con A receptors, visualized by Con A-HRP-DAB and Con A-biotin/avidin-ferritin techniques, were distributed uniformly on T. rhodesiense bloodstream forms. No lectin receptors were visualized on control preparations. Culture procyclics lacked a cell surface coat and were agglutinated by Con A and WGA but not RCA, SBA, PP and FBP. Procyclics were not agglutinated by lectins in the presence of competing sugar at 0.25 M. The expression of lectin binding cell surface saccharides of T. rhodesiense WRATat 1 is related to the parasite stage. Sugars resembling α-D-mannose are on the surface of bloodstream trypomastigotes and culture procyclics; n-acetyl-D-galactosamine and D-galactose residues are on bloodstream forms; and n-acetyl-D-glucosamine-like sugars are on procyclic stages.

Notes: The cationic permeant fluorescent dye rhodamine 123 (R123) was used to stain Plasmodium yoelii-infected mouse erythrocytes. Fluorescence microscopic observations demonstrated that the parasite, but not the matrix of the infected erythrocyte, accumulated the dye. Differences in fluorescence intensity could not be found at the various developmental stages of the parasite; however, quantitation of the cell-associated dye revealed an increase in R123 uptake with parasite development. The retention of the parasite-associated dye, as measured by fluorescence microscopy and spectrophotometry after extraction of R123 with butanol, was markedly reduced by treatment of the infected erythrocytes with a proton ionophore, carbonyl cyanide m-chlorophenylhydrazone (CCCP), and an inhibitor of proton ATPase, dicyclohexylcarbodiimide (DCCD). These results indicate that the accumulation and retention of R123 in P. yoelii reflect the parasite membrane potential and suggest that the parasite plasma membrane has a membrane potential-generating proton pump.

Notes: Distinctive organic-walled resting cysts of at least three different types with a highly conservative morphology appear to characterize specific orders or groups of genera within the Class Polyhymenophorea (Protozoa, Ciliophora), contrasting markedly with the great diversity of form seen in trophic stages. Polyhymenophorean ciliates have been considered in the past to form a cohesive class within the Phylum Ciliophora and, possibly, to represent the pinnacle of ciliate evolution. Evidence from cysts challenges the cohesive nature of the class, suggesting that the hypotrichs should be subdivided and that they have a different phylogenetic origin from the heterotrichs, tintinnids, and oligotrichs.

Notes: Yellow-brown, algal symbionts varying in diameter from approximately 5 μ m to 20 μ m, associated with solitary Radiolaria with spongiose skeletons (i.e. Spongodrymus sp.), exhibit fine structural features resembling the Prymnesiida (botanical class, Prymnesiophyceae). A large central vacuole is surrounded by a thin layer of cytoplasm containing plastids with lamellae composed of three thylakoids and granular pyrenoids with internal tubules immersed between the thylakoids. The pyrenoids lack internal thylakoid membranes. The nucleus is surrounded by a dilated cisterna of the nuclear envelope that also encloses the plastids and gives rise to saccules of the endoplasmic reticulum. The algal symbionts appear coccoid; hence no flagella nor surface scales were observed. The symbiont fine structure is compared to similar yellow-brown symbionts associated with Acantharia. Thus far, three kinds of algal symbionts have been observed to be associated with solitary Radiolaria: dinoflagellate, prasinomonad, and this apparent prymnesiomonad.

Notes: Ultrastructural observations of the cortically-located mitochondria of Tetrahymena thermophila revealed associations not only between the mitochondria and certain of the cortical microtubule bands, but also between the mitochondria and the epiplasm of the cortex. Most of the distal mitochondrial surface is close and parallel to the epiplasm; favorable views show bridge-like structures spanning the 20–10 nm gap between the mitochondrion and the epiplasm.Previous studies have shown that the placement of mitochondria in the cortex appears to be determined by certain of the cortical microtubule bands. This study, however, shows that mitochondrion-microtubule interactions account for only a small proportion of the total mitochondrial area associated with the cortex; the rest is accounted for by the epiplasm. A possible analogue of the spectrin layer of erythrocyte membranes, the epiplasm may be important in helping to arrange the intricately organized components of the ciliate cortex. Its involvement in apparently helping to “moor” mitochondria to their cortical sites is the first suggestion of any role in cell patterning played by the epiplasm.

Notes: Ce Tetradimorpha, rencontre en eau douce se présente soit sous forme sphérique pourvue de quatre flagelles et d'axopodes rayonnants, soit sous forme allongée avec a l'avant quatre flagelles associes a quatre axopodes et a l'arriére six a huit axopodes divergents. L'etude ultrastructurale révèle un cytosquelette axopodial de type centroplastidie comprenant un centroplaste lenticulaire homogéne, centre organisateur des quatre axopodes anterieurs et des six a huit axopodes posterieurs, auquel s'ajoutent les quatre cinetosomes des flagelles anterieurs. En outre, un deuxiéme éleément cytosquelettique incluant un microtubule associe chacun des quatre cinetosomes a l'axopode antérieur correspondant. Des cordons microfibrillaires réunissent axopodes et cinetosomes au niveau du centroplaste, puis a quelque distance du centroplaste les axopodes posterieurs. Les axonémes des axopodes comprenant de 5 a 30 microtubules sont constitues de triades, lorsqu'on peut détecter une organisation. Le noyau, a nucléole central est coince dans le cone axopodial posterieur, lui-méme entouré des dictyosomes. Par l'organisation du cytosquelette, par la structure des kinétocystes, par la structure des flagelles dépourvus de mastigonémes tubulaires, Tetradimorpha différe nettement de Ciliophrys marina. Comme le prévoyait Davidson (1975), il represönte bien un des chainons dans la série évolutive des Héliozoaires centrohélidiens. Mais il ne présente guère d'affinites avec les Chrysomonadines considerees comme la souche des Héliozoaires. L'intéret de ce Protiste dans l'étude de la differentiation et de l'evolution du cytosquelette est également présente.〈section xml:id="abs1-2"〉〈title type="main"〉ABSTRACTThis freshwater species of Tetradimorpha has a spherical body with four flagella and radiating axopods; it transforms into a pear-shaped cell that anteriorly has four flagella intercalated between four axopods and posteriorly has six to eight divergent axopods. Ultrastructural study reveals an axopodial cytoskeleton of the centrohelidan type comprising an homogeneous lenticular centroplast which acts as MTOC for axopodial microtubules. A second skeletal element is a microtubular linkage between the kinetosomes and the axonemes of anterior axopods. A microtubule embedded in dense material diverges from near the base of each kinetosomes and parallels the distal portion of the axoneme of each anterior axopod. A microfibrillar envelope around the centroplast links the axopodial bases to the kinetosomes situated just above. Close to the centroplast, microfibrillar strands link the axopodial axonemes to the kinetosomes. Axopodial axonemes are composed of 5 to 30 microtubules irregularly arranged except for some that form equilateral triangles. The nucleus containing a central nucleolus is constrained within a cone formed by the axonemes of the posterior axopods and surrounded by dictyosomes. By the cytoskeletal organization, the structure of kinetocysts, and flagella wthout tubular mastigonemes, Tetradimorpha differs obviously from Ciliophrys marina. As Davidson (1975) predicted, Tetradimorpha is an intermediate link in the centrohelidan lineage: however, it lacks the characteristics of chrysomonads, the supposed ancestors of Heliozoa. The contribution of this genus to the study of the differentiation and the evolution of the cytoskeleton is also presented and discussed.

Notes: Two-dimensional gel electrophoresis was used to identify the patterns of protein synthesis during initiation, and the patterns of membrane protein expression following initiation, in all of the mating types of the Tetrahymena thermophila B family. In addition, one-dimensional analysis was used to survey 125I-Concanavalin A-binding proteins. Although a large number of proteins was identified by each technique, no variation among the mating types was observed.

Notes: Cells of Tetrahymena pyriformis, T. thermophila, and Euglena gracilis were saturated with nitrogen gas at pressures up to 300 atm and rapidly decompressed. Damage was assessed by measuring post-decompression cell fragmentation or viability. Occurrence of intracellular bubbles was determined by cinephotomicrography performed during the decompression or by direct observations afterwards. The extreme gas supersaturations induced led to intracellular bubble formation and rupture in cells of Tetrahymena that contained food vacuoles, but only with supersaturations of 175 atm or higher; 225 atm left few cells intact. Bubbles were never observed in cells of Euglena or in Tetrahymena cells freed of food vacuoles, even when they were decompressed from substantially higher nitrogen supersaturations. Cells of Euglena were most resistant and were unaffected by supersaturations up to 250 atm.

Notes: The rapid, synchronous differentiation of N. gruberi from amoebae to flagellates is a useful paradigm to study aspects of cell differentiation, including regulation of the synthesis of proteins that are related to the changes in cell shape and motility, which occur during differentiation. The differentiation requires synthesis of new RNA and protein molecules to accomplish defined morphogenetic events. Specific new proteins, including the tubulins that form the flagellar microtubules, are synthesized at various times during differentiation, and particular mRNA species appear and disappear. The time course of the synthesis of the α and β subunits of flagellar tubulin is paralleled by the programmed appearance and disappearance of flagellar tubulin mRNAs. The evidence supports the hypothesis that the synthesis of flagellar tubulin is regulated by the transcription, and subsequent disappearance, of flagellar tubulin mRNA. Translatable mRNAs for two calmodulin-like calcium-binding proteins appear and disappear contemporaneously with those for flagellar tubulin. During differentiation the synthesis of actin, the major protein of amoebae, is selectively shut down, and translatable actin mRNA rapidly disappears. This description of the orderly appearance, utilization, and disappearance of the mRNAs for actin, calcium-binding proteins, and flagellar tubulin during differentiation provides means and motivation to investigate the mechanisms that regulate these events.

Notes: Earlier experimental work involving macronuclear implants in Stentor coeruleus has shown that the cytoplasmic cortex of the nuclear site 1) attracts the macronucleus and 2) holds it in place during interphase. Now experiments indicate macronuclei transferred with overlying cortex elongate in the direction of the transferred cortical pigment stripes, whether or not the transferred stripes realign in the direction of the host stentor's stripes. Therefore the third function of the cortex is to determine the direction of elongation and thus assure that both daughter cells at division receive part of the macronucleus.

Notes: During an electron microscopic study of Glugea stephani, three morphologically distinct tubular appendages that are continuous with the sporoblast plasmalemma were observed. The tubules were designated as: type I, 45–50 nm in diameter and 600–900 nm in length; type II, 25–35 nm in diameter, averaging 1300 nm in length; type III, 50–70 nm in diameter and with an indeterminate length, which often exceeds 3000 nm. Type III tubules contain regularly spaced, electron-dense particles that are approximately 30 nm in diameter. Since many genera of microsporida have some type of appendage, which may eventually be utilized for taxonomic purposes, we propose the formation of a system of serially numbered detailed descriptions of these structures to promote uniformity and clarity in future publications.

Notes: Cells of Paramecium tetraurelia, stock hrd, cultured in a micro-capillary containing 1 μl fresh culture medium, expressed mating activity through the whole cell cycle. Mating-reactive G2 phase cells can conjugate with cells of other phases. The G2 phase cells, which have double (4C) the normal micronuclear DNA content, undergo pre-meiotic DNA synthesis when conjugated with G1 phase cells. The micronucleus of the progeny from the cross between a G1 and a G2 cell becomes triploid.

Notes: Buffered solutions of KCl and NaCl were tested for their stimulatory effect on the germination of variously-aged spores of Vavraia culicis. Germination was optimal in 0.2 M KCl, pH 6.5 for one isolate, and, for another isolate, peaks of germination occurred at pH 7.0 and 9.5. Spores incubated for several hours in suboptimal solutions became unable to germinate under optimal conditions. After being returned to water, they regained their ability to germinate. Calcium chloride, magnesium chloride, and ammonium chloride inhibited germination. After ingestion by mosquito larvae, spores germinated near the posterior end of the midgut.

Notes: . Leishmania tropica promastigotes transport α-aminoisobutyric acid (AIB), the nonmetabolizable analog of neutral amino acids, against a substantial concentration gradient. AIB is not incorporated into cellular material but accumulates within the cells in an unaltered form. Intracellular AIB exchanges with external AIB. Various energy inhibitors (amytal, HOQNO, KCN, DNP, CCCP, and arsenate) and sulfhydryl reagents (NEM, pCMB, and iodoacetate) severely inhibit uptake. The uptake system is saturable with reference to AIB-and the Lineweaver-Burk plots show biphasic kinetics suggesting the involvement of two transport systems. AIB shares a common transport system with alanine, cysteine, glycine, methionine, serine, and proline. Uptake is regulated by feedback inhibition and transinhibition.

Notes: . Oxytricha strains used in biochemical studies have traditionally been grown in unaerated, unagitated culture tubes or Fernbach flasks. These cultures are limited in volume to about one liter and have a very nonuniform distribution of cells, with the majority of the cells at the very top or bottom of the medium. We have found conditions in which Oxytricha can be grown in 50-liter fermentation vats. The cultures grow to a uniform density of about 6000 cells/ml.

Notes: . The fine structure of the tomite of Foettingeria actiniarum (Claparède) was examined and compared with that of other apostome tomites. This stage in the life cycle has a unique configuration of kineties that form a spiral through the cytoplasm in the interior of the body. The structure and behavior of this internal spiral were evaluated as a mechanism for the storage of kinetosomes, an adaptation to the ciliate's two-host life cycle. The spiral is composed of nine ribbons of laterally compressed kinetosomes that are in contact with a thin electron-dense fibril. Paralleling the kineties of the spiral are conspicuous, swollen lamellae of the rough endoplasmic reticulum; these lamellae contain moderately electron-dense material. The spiral is associated with the large contractile vacuole and winds about the macronucleus. The tomite of Foettingeria possesses a single, robust, caudal cilium located in a pit, along with the nozzle-like pore of the contractile vacuole. The walls of the pit contain several trichocysts arranged radially about the caudal cilium and aimed into the pit.

Notes: . Fine structural studies of a specialized vesicle system associated with the endoplasmic reticulum (ER) of exo-erythrocytic Plasmodium berghei suggest that this system may be the equivalent of a Golgi apparatus. Patches of ER, randomly distributed in the cytoplasm of developing parasites, are formed of smooth and ribosome-studded cisternae intermingled with each other. The vesicle systems are located between as well as at the edges of ER aggregates and appear to be in different stages of budding from the cisternae. Prolonged osmication reveals distinct staining of the nuclear envelope and ER of the parasites as well as part of the Golgi apparatus of the hepatocytes. However, the small vesicles associated with the parasite's ER are unstained, as are the coated vesicles in the Golgi region of the liver cell. These sites in the parasite cytoplasm seem comparable to the concave surface of the Golgi apparatus in liver cells. The pinched-off vesicles fuse with others to form the prominent peripheral vacuolization characteristic of the nearly mature exo-erythrocytic form. The formation of these peripheral vacuoles and their subsequent fusion with the parasite membrane may be an exocytosis mechanism supplying the rapidly expanding parasite with new plasma membrane material.

Notes: . Ultrastructural cytochemical techniques were used to analyze the nucleus and the kinetoplast of epimastigotes of Trypanosoma cruzi. With the use of ethanolic phosphotungstic acid, which detects basic proteins, reaction product was seen in the chromatin and at the periphery of the kinetoplast. Thallium alcoholate, which interacts with DNA, stained strongly the whole kinetoplast and the chromatin. With the use of a silver impregnation method that detects acidic nucleolar proteins, silver granules were seen preferentially located in the central region of the nucleolus. With the EDTA method, which reveals the presence of ribonucleoproteins, staining was observed in the nuclear pores. Also 6–8 nm fibrils, 25 nm and 40 nm granules, which correspond to the perichromatin fibers, interchromatin granules and the perichromatin granules, respectively, were identified in the nucleus. The EDTA method also revealed the presence of 40 nm granules in the kinetoplast. These granules were seen mainly at the two extremities of the kinetoplast. Freeze-fracture images indicate that the nuclear membrane contains ca. 9 pores/μm2 of nuclear surface area. The mean diameter of the pores was 80 nm. All these results suggest that epimastigotes of T. cruzi have a very active nucleus and a high rate of nucleocytoplasmic interchange.

Notes: Sorogena stoianovitchae is an unusual ciliated protozoan with a life cycle characterized by the aggregation of individual trophic cells to form a multicellular sorogen that rises from the liquid culture medium surface by the secretion of a stalk. The noncellular stalk is a tapered, longitudinally furrowed structure composed of a fibrillar matrix that is initially hydrated, but with time dehydrates, the stalk becoming thin and brittle. This dehydration is of importance from the earliest stages of stalk formation since it results in the formation of the outer sheath-like region of the stalk that appears to provide much of the support of the stalk. Cytochemical tests of the stalk for polysaccharides (including acidic mucopolysaccharides) and proteins are positive. Proteolytic enzymes degrade the stalk. Lectins specific for glucose and N-acetyl-D-glucosamine bind to the stalk. Gas chromatography analysis detected the presence of fucose, glucose, glucosamine, and arabinose, as well as a variety of amino acids, predominantly glycine. The cytochemical and biochemical tests, the ultrastructural data, and the behavior of the stalk material suggest that the staik is composed of a matrix of complex protein-polysaccharide molecules.

Notes: Nosema disstriae, a parasite of the forest tent caterpillar, Malacosoma disstria, was cultured with cell lines UMN-MDH-1 (Malacosoma disstria), IPLB-1075 (Heliothis zea), and BTC-32 (Triatoma infestans). Infected cultured cells were used to infect the healthy cell lines. Electron micrographs of thin sections of 6-day-old cultures revealed infected cells that exocytosed vesicles containing vegetative and immature sporulating forms of the parasite. Some of these forms were believed to be responsible for intercellular transmission of the parasite. The spread of infection was augmented by culturing the cells at high densities; if the density was too low, there was little or no cross infection. Cross infection was inhibited, but not blocked completely, by high osmolality of the culture medium. The yield of spores from a confluent cell monolayer at the end of growth was generally 1–4 × 107 per ml of culture medium.

Notes: The fine structure of the trophozoite of Acanthamoeba palestinensis with a special emphasis on the Golgi complex, microbodies, and mitochondria has been examined. Golgi complexes are distributed throughout the cytoplasm but are most abundant in the perinuclear region. Usually two Goigi complexes are found in the same plane on opposite sides of the nucleus. One of them appears to be in an intimate association with the nuclear membrane. The region of contact contains compact cisternae, vesicles of various sizes, as well as granular and amorphous electron-dense material. Structural changes in the nuclear envelope are also observed in this area. A structure consisting of a Golgi complex and electron-dense microtubule organizing center, comparable to the centrosphere of other Acanthamoeba species, has been observed. Microbodies, surrounded by a single unit membrane and containing a granular matrix and tubular inclusions, are scattered throughout the cytoplasm. These organelles, circular (∼1 μm in diameter) or ovoidal (∼1 μm in length and ∼0.5 μm in width) in section, have often an irregular outline. These microbodies are probably the morphological equivalent of peroxisomes and glyoxysomes. Most mitochondria show a typical structure including tubular cristae and intracristal inclusions. Occasionally mitochondria with two apposed double membranes running through the midline are found. Such atypical cristae have never been reported in small amoebae before.

Notes: Heavy infections with enigmatic mobile organisms have recently been found in the blood of carp (Cyprinus carpio) in Central Europe. The organisms measure up to 15 μm, are variable in shape, and exhibit an unceasing twitching or dancing movement. Their developmental cycle starts with a primary cell enclosing a secondary cell. The former grows while the latter produces inside itself by a series of binary fissions and internal cleavages up to eight secondary cells, each of which encloses an inner (tertiary) cell of its own. In addition, up to four tiny cells with compact nuclei (“residual bodies”) also result from divisions of the secondary cells. Primary cells containing the products of the division of secondary cells finally disintegrate, releasing the secondary cells, which in their turn become new primary cells and repeat the cycle all over again. The structure and behavior of these organisms were so incompatible with existing ideas on myxosporean development that their myxosporean affinity was at first unrecognized. The final proof of their identity–appearance of myxosporean spores in sterile, experimentally infected hosts–is still to be presented. The interpretation of the myxosporean features of their life cycle (i.e., [1] the pericyte nature of the primary cell, [2] proliferation by disintegration of the pseudoplasmodial primary cell, [3] no rigidly fixed pattern in vegetative development), their ultrastructure (i.e., [1] characteristic bundles of microtubules and numerous free ribosomes in secondary cells, [2] lack of centrioles, [3] membranes enclosing the secondary cells within the primary cells), and facts on their epizootiology (i.e., [1] no success at transmission via leeches, [2] the occurrence of these organisms along with Sphaerospora renicola Dykova and Lom) suggest that they are stages of S. renicola from the kidney of carp. Similar mobile organisms were found in the blood of fry of two other fishes (Gobio gobi and Tinca tinca) which are also hosts for a Sphaerospora that infects the kidney. This suggests that these organisms represent an early phase in the developmental cycle in the genus Sphaerospora. The existence of cells enveloped one within the other (secondary and tertiary cells) in the developmental cycle, a characteristic myxosporean feature itself, is an intriguing parallel to similarly enclosed cells in sporogenesis of Paramyxea (Ascetospora).

Notes: The morphology and morphogenesis of the kinetofragminophoran soil ciliates, Fuscheria terricola n. sp. and Spathidium muscorum Dragesco & Dragesco-Kerneis, 1979, are described. Stained specimens (protargol) are characterized biometrically. The new species differs from the other species of the genus in its body size, body shape, number of kineties, length of extrusomes, and habitat. Both species have telokinetal stomatogenesis, which commences with a proliferation of kinetosomes at those kineties which bear the brosse. Fuscheria terricola does not have a complex perioral ciliature; indeed, it might be that this species has only monokinetids. Thus only a proliferation of kinetosomes and the separation of the kineties takes place in the prospective division furrow. In contrast, S. muscorum differentiates short dikinetid kinetofragments in the region of the division furrow, which are arranged to form the perioral kinety of the opisthe in the intermediate and late stages of the stomatogenesis. The right part of the perioral kinety develops first. This and other studies show that telokinetal stomatogenesis proceeds very differently depending on the differentiation of the oral ciliature; however, detailed studies on the morphogenesis of kinetofragminophoran ciliates are still too few in number for subtypes to be defined.

Notes: The apicomplexan family Barrouxiidae Léger, 1911 is reviewed and revised on the basis of present information. It includes the genera Barrouxia Schneider, 1885 with ten named species, Defretinella Henneré, 1966 with one named species, and Goussia Labbé, 1896 with 25 named species. The family is characterized by having bivalved sporocysts with a longitudinal suture line. Available information, admittedly spotty, is given for each species on oocyst, sporocyst and sporozoite structure, and on locus of sporulation. The following seven new combinations are made: Goussia flaviviridis (Setna & Bana, 1935) n. comb. in the gecko Hemidactylus flaviviridis; G. hyalina (Léger, 1898) n. comb. in an unidentified aquatic beetle; G. lacazei (Labbé, 1895) n. comb. in the centipedes Lithobius forficatus and L. martini; G. metchnikovi (Laveran, 1897) n. comb. in the gobies Gobio gobio and G. albipinnatus; G. schaudinniana (Pinto, 1928) n. comb. in the centipede Lithobius forficatus; G. stankovitchi (Pinto, 1928) n. comb. in the small bleak Alburnus alburnus, the bream Abramis brama, and the red roach Scardinius erythrophthalmus; Goussia sp. (Dogel' Akhmerov, 1959) nov. comb. in the freshwater fish Gnathopogon chankaensis.

Notes: Comparison of the electrophoretic migration patterns of proteins of active 40S and 60S ribosomal subunits isolated from nine amicronucleate strains of Tetrahymena of known phenoset revealed strain dependent differences which correlated with the phenoset classification of these strains as determined by Borden, Whitt & Nanney, who compared isoenzyme patterns.

Notes: Chromatin from a uninucleate dinoflagellate, Crypthecodinium cohnii, a binucleate dinoflagellate, Peridinium balticum, and a chromophyte, Olisthodiscus luteus, was examined by nuclease digestion and the results were compared to those from vertebrates. Gel analysis of the products of staphylococcal (micrococcal) nuclease digestion revealed a DNA repeat unit of 220(±5) base pairs for O. luteus and 215(±5) for P. balticum. Limit digestion gave a core particle of 140 base pairs, revealing that these longer repeat sizes are due to longer linker regions. No repeating subunit structure was found upon electrophoresis of digests of C. cohnii nuclei. Examination of the DNA fragments produced by DNAse I digestion of nuclei isolated from P. balticum and O. luteus showed the same ladder of ten base multiples as seen in chromatin from other eukaryotes. Examination of the kinetics of digestion by DNAse II of Peridinium chromatin revealed less susceptibility when compared to DNAse I digestions while 70% of Olisthodiscus chromatin and 35% of C. cohnii chromatin was sensitive to DNAse II. These data, taken together with previous results from Euglena, indicate that while algal chromatin is similar to that of higher eukaryotes in regard to DNAse I and II action, it differs in that the linker DNA is longer. In addition, the Hl-like histone from O. luteus and P. balticum is located in the linker DNA as in higher eukaryotes.

Notes: Changes in mean cell size, DNA and cell density were monitored at 6-h intervals for 72 h in populations of six species (eight clones) of marine dinoflagellates to determine the temporal relationships between the cell cycle events of DNA replication and cytokinesis. Batch cultures were maintained at 15 or 20°C on a 12-h light: 12-h dark photoperiod. Cell densities and size frequency distributions were determined conductimetrically and the amount of DNA within populations was measured fluorometrically. A variety of intra- and interspecific relationships were observed, ranging from parallel phasing of cell cycle processes to variations which involved the temporal uncoupling of DNA synthesis from the phased pattern of cell division which is characteristic of dinoflagellate cell cycles. Daily growth rates of individual populations varied from 0.05 (Gymnodinium nelsoni) to 2.08 (Amphidinium carteri) cell divisions day-1 and DNA doubling rates ranged from 0 to 1.14 day-1. Mean doubling rates for DNA were usually 30–40% lower than those for cells. The degree of difference in these rates and the amount of variability evident in cell cycle sequences may be major factors in determining the rate and extent of development of dinoflagellate populations in nature.

Notes: In Aedes cantator, Amblyospora sp. is transovarially transmitted and has two developmental sequences. The life cycle is initiated in the adult female with the release of sporoplasms from binucleated spores not bounded by membranes, lying free within host oenocytes. Sporoplasms infect the developing oocytes and are transmitted to the filial generation when the eggs are laid. In some of the female progeny that hatch from infected eggs, diplokaryotic cells infect host oenocytes and divide by binary fission during merogony. Sporulation and spore formation do not occur until a blood meal is taken by the host and they coincide with the development and maturation of the oocytes to complete the cycle. In other female and all male progeny, pathogen development occurs within fat body tissue of the host where diplokaryotic cells divide by multiple fission during merogony to spread the infection. Sporulation in this developmental sequence is characterized by the secretion of an accessory membrane and the meiotic division of diplokaryotic sporonts, which result in the formation of octonucleated plasmodia that undergo cytokinesis to form eight haploid spores which are not perorally infectious to other mosquito larvae. There is no increase in the prevalence of either type of infection in field populations during juvenile development, indicating that there is no direct horizontal transmission of the pathogen within any one generation. Data obtained from laboratory rearings of infected progeny, however, show that infections cannot persist relying solely upon maternal-mediated transmission and that some other mode of transmission must be operative for continued maintenance of this microsporidium in A. cantator.

Notes: Cells of Amoeba proteus and Chaos carolinensis that were in the process of phagocytosing large prey organisms were studied to find a structural basis for the generation of mechanical forces exerted by newly forming food cups. It was found that the food-cup walls facing prey organisms have a more prominent network of thin filaments inside the plasmalemma and that the glycocalyx covering the area is more condensed than usual.

Notes: Monoclonal IgG antibodies against sporozoites of Eimeria tenella were obtained from the ascites fluid of BALB/c mice. Oocysts, sporocysts, and sporozoites were exposed to medium 199, normal ascites fluid, or monoclonal antibodies 1A, 9D, 3D3II, or 2G8f. Specimens were then incubated with ferritin-conjugated goat anti-mouse IgG antibody. Ferritin was uniformly distributed over the surface of sporozoites exposed to 1A, 9D, or 3D3II; ferritin was localized in patches on sporozoites exposed to 2G8f. A uniform layer of ferritin was present on the inner layer of oocyst walls and on the Stieda body and outer surface of sporocysts exposed to 1A, 9D or 3D3II. In specimens treated with 2G8f, ferritin was present on the inner layer of the oocyst wall and the Stieda body, but not on the sporocyst wall. No ferritin was found on specimens exposed to medium 199 or normal ascites fluid. Monoclonal antibodies 1A, 9D, and 3D3II, but not 2G8f, caused complement-mediated lysis of sporozoites. These findings indicate that oocysts, sporocysts, and sporozoites of E. tenella contain common antigens specific for each monoclonal antibody tested.

Notes: Several divalent cation-dependent ATP phosphohydrolases associated with cilia, ciliary axonemes, ciliary membranes, pellicles, trichocysts, nuclei, mitochondria, microsomes, and soluble peripheral cell surface fractions of Paramecium tetraurelia were resolved in this study. Fifteen different activity bands were detected in whole cell sonicates or subcellular fractions by Triton polyacrylamide gel electrophoresis and ATPase activity staining. The ciliary surface membrane contained two major ATPase activities that were distinct from the enzymes associated with all other cell fractions.

Notes: During feeding a peritrophic membrane (PM) is formed in the gut of the tick Ixodes dammini, dividing the lumen of the gut into an ecto- and endoperitrophic space. Babesia and all food particles ingested with the blood meal by the tick are retained in the endoperitrophic space, the lumen proper. Only Babesia equipped with a highly specialized organelle, the arrowhead, are able to pass the PM and enter the ectoperitrophic compartment. During the crossing of the PM the arrowhead loses its density, suggesting that enzymes released from it dissolve the polymers in the PM, making passage of the parasite through this barrier possible. In the ectoperitrophic space the arrowhead of Babesia touches the epithelial cell. At the point of contact the membrane of the host cell starts to invaginate, and simultaneously the arrowhead's fine structure loses its highly organized pattern. The growing host membrane encircles the parasite and the arrowhead diminishes progressively in size. When the piroplasm is inside the host cell, the arrowhead can no longer be found. During invasion the host membrane often touches the parasite's plasma membrane at the site of a coiled structure, and the host membrane becomes ruptured and the nearby host cytoplasm appears to be lysed. Babesia inside the host cell is covered solely by its own plasma membrane; the invaginated host membrane is missing. It is postulated that the latter disintegrates during invasion by the parasite through the action of enzymes from the coiled structure. The parasite is surrounded by a halo of homogeneous material deriving most probably from the lysed host cytoplasm.

Notes: A technique for the separation of schizonts of Plasmodium falciparum is described. The different stages of the asexual cell cycle of the parasite were positioned according to their density in a continuous gradient of Percoll. Young trophozoites coincided with erythrocytes in a broad band corresponding to densities from 1.075 to 1.100 g/ml, whereas schizonts were concentrated at a density approximating 1.062 g/ml. The viability of the parasites was unimpaired by this procedure. Young trophozoites and schizonts continued their normal life cycle when cultured after the separation procedure. The percentage of recovery was high, reaching 80% of the initial quantity. Possible applications of the technique are discussed.

Notes: Chemically defined minimal media for the cultivation of high temperature tolerant and pathogenic Naegleria spp. have been developed. A defined minimal medium, identical for N. fowleri and N. lovaniensis, consists of eleven amino acids (arginine, glycine, histidine, isoleucine, leucine, methionine, phenylalanine, proline, threonine, tryptophan, and valine), six vitamins (biotin, folic acid, hemin, pyridoxal, riboflavin, and thiamine), guanosine, glucose, salts, and metals. Three of the four strains of Naegleria fowleri tested (ATCCr̀30100, ATCCr̀30863, and ATCCr̀30896) and two strains of N. lovaniensis (ATCCr̀30467 and ATCCr̀30569) could be cultured beyond ten subcultures on this medium. For N. fowleri ATCCr̀30894 diaminopimelic acid, or lysine, or glutamic acid was also required. Mean generation time was reduced and population density increased for all strains with the introduction of glutamic acid. Glucose could be eliminated from the minimal medium only if glutamic acid was present. Without glucose, mean generation time increased and population density decreased. Diaminopimelic acid could substitute for lysine for ATCCr̀30894, indicating that Naegleria species may synthesize their lysine via the DAP pathway. Naegleria fowleri ATCCr̀30100 could be adapted to grow without serine or glycine in the minimal medium with glutamic acid added, but with mean generation time increased and population density decreased. The strain could be grown in the minimal medium in the absence of metals. For growth of N. australiensis ATCCr̀30958, modification of the medium by increasing metals ten-fold, substituting guanine for guanosine and adding lysine, glutamic acid, and six vitamins (p-aminobenzoic acid, choline chloride, inositol, vitamin B12, nicotinamide, and Ca pantothenate) was required.

Notes: From several surveys of environmental sites, the virulent human pathogen, Naegleria fowleri, was isolated from a pond in Georgia, a sewage treatment plant in Missouri, and from the Potomac and Anacostia rivers near and in Washington, D.C. Widely scattered, sparse populations seemed only a potential threat to human health at the time of sampling. The data support an estimate that the sites sampled contain 10,000 typical, low temperature, bactivorous amoebae for each heat tolerant amoeba able to grow at 45° C. Heat tolerant competitors were much more common than N. fowleri. Naegleria lovaniensis, which is heat tolerant but nonpathogenic, was isolated from and downstream from an open air thermal pollution temperature gradient. Hot piles of composting sewage sludge yielded no amoeboflagellates, many heat tolerant (45–49° C) amoebae, and one thermophilic (52° C) Acanthamoeba. Features of the methods used include two-stage incubation to increase isolation of sparse organisms and distinction of N. fowleri from almost all other amoebae on agar plates. The flagellate-empty habitat hypothesis postulates a general model in which human intervention and/ or natural events remove usual competitors and the ability to transform to a motile flagellate confers an advantage in recolonizing.