ALBERT

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: We examined the long-term success of prairie planting on a former strip mine in northeastern Illinois. The site was reclaimed and planted with prairie species in the 1970s. Total biomass increased over time, largely as a result of an increase in biomass of non-prairie species. Biomass of prairie species remained unchanged because of an increase in Panicum virgatum (switchgrass) offsetting decreases in Sorghastrum nutans (Indian grass). Total biomass was less than values published for other restored prairies (78 ± 4 g/m2to 298 ± 72 g/m2 for our site, as opposed to 302-489 g/m2 for the Trelease Prairie). Mycorrhizal inoculum potential (MIP) was variable across the site. There were also relatively few species of mycorrhizal fungi present as spores. Gigaspora sp., Scutellospora sp., Glomus sp., Glomus geosporum, and Glomus cf. fasciculatum were identified from spores. On a transect dominated by warm-season (C4) prairie grasses, MIP of rhizosphere soil collected under these species was lower than the MIP of rhizosphere soil collected under exotic cool-season (C3) grasses on a transect dominated by C3 species. On a transect with mixed warm-and cool-season vegetation, however, MIP did not differ under the two vegetation types. These results suggest that within-site patchiness rather than cover type is influencing MIP. Values of MIP were lower than those reported for native Illinois prairie.

Notes: Twenty-three “vital ecosystem attributes” (VEAs) were previously proposed to aid in quantitative evaluation of whole ecosystem structure, composition, and functional complexity over time. We here introduce a series of 16 quantifiable attributes for use at a higher spatial scale and ecological organizational level, the landscape. “Vital landscape attributes” (VLAs) should be useful in evaluating the results of ecological restoration or rehabilitation undertaken with a landscape perspective, provided that clear definitions and boundaries are agreed upon for the different spatial and ecological entities involved. Like VEAs, VLAs should be sensitive to changes wrought by human as well as to nonhuman factors leading to ruptures in flow processes or vegetation “switches.” They should be applicable over a wide range of landscape types and therefore aid in conducting rigorous interlandscape comparisons. We present three groups of VLAs: (1) landscape structure and biotic composition, (2) functional interactions among ecosystems within the landscape, and (3) degree, type, and causes of landscape fragmentation and degradation. Ecotones between ecosystems are touched upon by several different VLAs. Because conflicting terminology abounds in this area, we append a glossary defining the problematic terms used.

Notes: Heightening human impacts on the Earth result in widespread losses of production and conservation values and make large-scale ecosystem restoration increasingly urgent. Tackling this problem requires the development of general guiding principles for restoration so that we can move away from the ad hoc, site- and situation-specific approach that now prevails. A continuum of restoration efforts can be recognized, ranging from restoration of localized highly degraded sites to restoration of entire landscapes for production and/or conservation reasons. We emphasize the importance of developing restoration methodologies that are applicable at the landscape scale. Key processes in restoration include identifying and dealing with the processes leading to degradation in the first place, determining realistic goals and measures of success, developing methods for implementing the goals and incorporating them into land-management and planning strategies, and monitoring the restoration and assessing its success. Few of these procedures are currently incorporated in many restoration projects. The concept that many ecosystems are likely to exist in alternative stable states, depending on their history, is relevant to the setting of restoration goals. A range of measures, such as those being developed to measure ecosystem health, could be used to develop “scorecards” for restoration efforts. Generalizable guidelines for restoration on individual sites could be based on the concepts of designed disturbance, controlled colonization, and controlled species performance. Fewer explicit guidelines are available at the landscape scale, beyond nonquantitative generalities about size and connectivity. Development of these guidelines is an important priority so that urgent large-scale restoration can be planned and implemented effectively.

Notes: Seagrass ecosystems fulfill ecologically and economically valuable functions in coastal marine environments. Unfortunately, seagrass beds are susceptible to natural and human disturbances, and their distrubution is declining worldwide. Although intentional disturbance of seagrass beds must be mitigated pursuant to U.S. law, to date mitigation of seagrass beds has not prevented a net loss of habitat. Transplantation of vegetative material from small areas of nearby beds is the primary method of seagrass mitigation. Restoration research on seagrasses has focused primarily on establishment of the plants and secondarily on the functional equivalency of the habitats. We questioned whether transplanted seagrass beds were comparable to “natural” beds in terms of genetic diversity and structure. We sampled Zostera marina L. (eel-grass) from 12 sites in the highly urbanized area of San Diego County and from pristine sites in Baja California. Using allozyme electrophoresis, we determined that genetic diversity (percentage of polymorphic loci, allele richness, expected and observed heterozygosities, and proportion of genetically unique individuals) was significantly reduced in transplanted eelgrass beds. Eelgrass from Baja California exhibited the highest genetic diversity. Based on Wright's F statistics, most of the genetic variation was distributed within rather than among sites (FST= 0.139), and the degree of genetic structure was only moderate at the greatest geographical scale (San Diego—Baja). Using a spatial statistical analysis (second-order analysis), we found virtually no evidence for nonrandom distribution of alleles or genotypes at scales of 3–50 m within beds. We discuss several hypotheses for reduced genetic diversity in transplanted eelgrass beds, including transplantation protocol, small size of transplantations, and reduced or failed sexual reproduction.

Notes: Few early examples of forest restoration projects are extant in the formerly forested parts of eastern North America. In this paper I present the history and status of an early forest restoration project in a denuded gravel pit in Ontario, Canada. The site was part of a deciduous forest until 1840, at which time forest clearing occurred. From 1874 to 1886 the site was exploited as a gravel pit. In 1887, under the direction of William Brown, it was planted with 14 species of coniferous and deciduous trees, of which 10 are still present. No soil preparation was carried out. The trees were pruned for 7 years, but in 1892 intensive maintenance ceased. In the 107 years since planting, the site has acquired some structural characteristics similar to the surrounding native deciduous forest, but it retains characteristics of an artificial community. Canopy cover has increased from 85% to over 95% since 1930 and is primarily Juglans nigra and Acer platan aides, Survivorship and current growth rates of native and exotic taxa have been similar. Recruitment patterns suggest that J. nigra and A. platanoides will dominate the canopy over the next century. The site is a useful example of the progress and problems created by attempts at forest restoration today.

Notes: We monitored several herbaceous species for revegetating motorway slopes in Catalonia, Spain, a Mediterranean country. Two main kinds of treatments were applied: hydroseeding on bare marl, chalk, and slates, and hydroseeding on embankments over gentler slopes, where soil materials previously removed were spread before hydroseeding. The greatest herbaceous cover was obtained by hydroseeding after soil replacement, and marl was the most suitable bare substratum for hydroseeding. Physical characteristics such as schistosily plane in slates and softness or surface irregularity in chalk determined the outcome of revegetation efforts. The most immediate stabilization of soils was obtained on southern exposures with autumnal applications. Grasses, markedly of the genera Lolium and Festuca, were dominant in the herbaceous cover at the end of the monitoring period. Natural invasive vegetation was composed of ruderal species, but no representatives of the adjacent forest or maquis community were found.

Notes: Books reviewed in this article: Restoration Ecology in Europe Krystyna M. Urbanska and Krystyna Grodzinskn, editors Ecology and Management of Invasive Riverside Plants Louise C. de Wnal, Lois E. Child, P. Max Wnde, and Joint H. Brock, editors Restoring Prairie Wetlands: An Ecological Approach Susan G. Galatowitsch and Arnold G. van der Valk

Notes: A rehabilitation procedure designed to reestablish resource control processes in a degraded Acacia aneura woodland was successful in improving soil nitrogen and carbon content, exchange properties, and water infiltration rates. Soil respiration rates and soil fauna populations increased, and soil temperatures were moderated. The procedure comprised laying piles of branches in patches on the contour of bare, gently sloping landscapes, with the expectation that soil, water, and litter would accumulate in these branch piles, thus improving the soil habitat and its productive potential. The procedure was derived from landscape function analysis, indicating that surface water flow was the principal means of resource transfer in these landscapes. Under degradation such overland flow results in a loss of resources. This rehabilitation procedure reversed loss processes, resulting in gains in the productive potential of soils within patches. This procedure was successful despite grazing pressure being maintained throughout the experiment.

Notes: Ursus americanus(black bear) predation could limit the success of the proposed restoration of Rangifer tarandus (woodland caribou) to Minnesota. The problem was recently identified as a major factor in the failure of a similar restoration effort in Maine. During the summer of 1991 we conducted a survey in the region of the proposed restoration, using bait stations to identify bear presence. Four settings were sampled: islands with campsites, islands without campsites, mainland areas with campsites, and mainland areas without campsites. Results from the survey suggest that black bears use areas with campsites more than those without. Whereas caribou may use islands preferentially for calving to escape predation, islands with campsites may be unfavorable for caribou calf survival due to frequent bear visitation.

Notes: The past 40 years have seen the legal and policy framework for nature conservation in Britain extend from protection and preservation to include enhancement through techniques such as ecological restoration and habitat creation. Clear objectives need to be set for ecological restoration and habitat creation schemes because the processes involve human intervention in combination with natural factors operating over time. Objectives are required for both management and monitoring in order to enable measurement of success or failure. The most effective way to achieve high-quality restoration and creation schemes is to define the output of the process—a habitat, vegetation type, or biological community. The better the definition of the output, the greater the need to define the inputs and the nature of the intervening processes. Ecologists and environmental managers have a key role to play in establishing the degree of definition necessary in order to achieve a particular objective.

Notes: Whetstone Brook is a trout stream located in north-central Massachusetts that is degraded by acid precipitation. The stream was treated with 56 tonnes of powdered limestone by a prototype, water-powered doser as part of the U.S. Fish and Wildlife Service's Acid Precipitation Mitigation Program. The goal of liming Whetstone Brook was to raise the pH to 6.5 and acid neutralizing capacity (ANC) to at least 50 μeq/1 in a 3.2-km reach. This goal was achieved despite the fact that during the 31 months of treatment stream flow was 37% higher than during the pretreatment period. During the treatment period, pH averaged 6.54 and ANC averaged 69.75. During the pretreatment period average pH was 5.97 and average ANC was 20.26. In the control section of Whetstone Brook, both pH and ANC were lower during the treatment period than during the pretreatment period. During treatment, monomeric aluminum, a form toxic to fish, declined in the treated section and increased in the control section. Total calcium, sediment calcium, and pore-water calcium increased in the treated section during treatment but declined in the control section. The other base anions and cations, nutrients, and physical parameters were not significantly affected by liming.

Notes: Books reviewed in this article: Redesigning the American Lawn: A Search for Environmental Harmony. F. H. Bormann, D. Balmori, and G. T. Geballe Seasonally Dry Tropical Forests Stephen H. Bullock, Harold A. Mooney, and Ernesto Medina, editors

Notes: A limestone slurry was sprayed on the surface of Thrush Lake, a small headwater lake in northeastern Minnesota, to test a treatment designed to protect acid-sensitive waters from anthropogenic acidification. The 6-year study, consisting of pretreatment, transition, and post-treatment phases, was part of the four-state Acid Precipitation Mitigation Program directed by the U.S. Fish and Wildlife Service. Measured water-chemistry parameters, including acid-neutralizing capacity, pH, dissolved calcium, and dissolved inoroganic carbon, increased following treatment, although local climatic conditions influenced the magnitude and duration of the chemical changes. Physical changes to the lake, other than an increase in conductivity and a short-term alteration of water clarity subsequent to treatment, were not documented. The composition of the zooplankton community was altered, with the proportion of rotifers increasing after treatment. Individual zooplankton species showed a variety of changes in abundance that were associated with treatment over both seasonal and multi-year intervals. For example, Holopedium gibbemm was absent from lake samples immediately following treatment and recovered within a season, whereas Diaptomus minutus and Keratella taurocephala populations were reduced after treatment and had not recovered by the end of the study. Alternately, Asplanchna priodonta increased in abundance after treatment. These observed abundance patterns were generally consistent with previzous acidification or base-addition studies. In contrast, the changes in community composition of zooplankton did not consistently fit patterns developed from regional studies across water-chemistry gradients. These differences emphasize the importance of biotic as well as abiotic factors in controlling zooplankton community composition.

Notes: The restoration of chemically degraded rivers, lakes, and estuaries with large watersheds and pollution sources that are primarily diffuse in nature requires the grading of thousands of kilometers of tributary streams. Many population-and community-oriented biomonitoring methods have been developed that avoid the cost limitations of chemical/biomarker/bioassay approaches and the serious limitations of single-factor analysis as related to complex systems. In this study of the coastal plain and piedmont geomorphologic provinces of the Chesapeake Bay watershed, we have demonstrated a set of quantitative measures based on analysis of macrophyte populations that provide statistically significant separation of streams in accordance with their state-issued water quality rating. Macrophytes can be abundant and diverse in lower-order streams, and they demonstrate patterns of community structure and diversity similar to those of other organisms developed for biomonitoring of stream degradation. Unlike organisms previously and extensively used in biomonitoring techniques, however, macrophytes are considerably easier to identify and quantify. In addition, macrophyte techniques provide a range of measures of increasing sensitivity from species numbers at a few sites, to the presence/absence and abundance of indicator species, and, finally, to a diversity analysis based on easily identified species at an extended number of sites. We suggest that the ease of utilization of this methodology will allow repeated surveys of all streams in large watersheds with the invertebrate, fish and diatom biomonitoring to biomarking and chemical bioassays and finally analytical chemistry, progressively applied to verify and then identify specific pollution sources (“hot spots”) in a more limited number of problem streams.

Notes: Alliaria petiolata (garlic mustard), an exotic plant species, has invaded woodlands in several areas in mid-western and northeastern United States and adjacent Canada, and it is displacing the indigenous under-story flora. This study was conducted to provide information about the species' biology that might be useful in controlling its spread in native woodlands. The plant is a strict biennial in North America, spending the first year of growth as a basal rosette. This period of relatively slow growth is followed by a period of rapid shoot elongation (1.9 cm/day) during early spring of the second growing season. The plant is capable of cross- and self-pollination, although pollination and stigma receptivity occur before the flower is open, so autogamy is the most likely breeding system. Garlic mustard invests 20.4% of its biomass in reproductive effort, with an annual seed rain of 15,000 seeds/m2. Seed dispersal from fruits begins in early July and continues into October. Most seeds germinate in the spring following the year in which they were produced. Seedling recruitment is high (8.3–18.0 seedlings/cm2), but only about 7.5% of the plants survive to maturity. The success of the plant in invading woodlands appears to be related to (1) its autogamous breeding system that allows a single individual, or a few individuals, to establish populations of genetically similar but interfertile individuals; (2) high seed production, permitting establishment of large numbers of individuals; and (3) rapid growth during the second growing season, which increases its competitive ability. Because of garlic mustard's ability to occupy understory habitats successfully, it may be unrealistic to expect to eliminate the plant from many habitats it has already invaded.

Notes: This study investigates the influence of gap width and turf type on the growth of planted seedlings of the Australian forb Bulbine bulbosa (bulbine lily) and subsequent recruitment of this species from self-sown seed. In a low-productivity turf of Danthonia setacea (bristly wallaby grass), planted Bulbine seedlings established satisfactorily in all gaps 50 mm wide or larger. In highly productive turfs of Festuca arundinacea (tall fescue) successful, establishment and growth of planted seedlings required a competition-free gap more than 200 mm wide. Successful recruitment of self-sown Bulbine seedlings was observed in all gap widths in Danthonia turfs. In Festuca, however, seedling recruitment was low, irrespective of gap width. The results are related to establishing Bulbine bulbosa in habitat reconstruction programs in southeastern Australia.

Notes: We conducted a study of the flood tolerance of nine wetland tree species on seven soil types. Seedlings were subjected to 11 months of continuous shallow inundation or moist soil conditions on three mineral soils, two organic soils, a manufactured soil designed to mimic the practice of layering muck over mineral soil, and a stockpiled topsoil. Taxodium ascendens, T. distichum, Acer rubrum, and Pinus serotina suffered no mortality; Fraxinus carolininna (1%), Liquidambar styraciflua (8%), P. elliottii (8%), and Gordonia lasianthus (24%) suffered low to moderate mortality; and Persea palustris (46%) suffered significant mortality. In general, greatest net height and total biomass were achieved on moist organic soils, and least net height and total biomass were achieved on stockpiled topsoil and inundated soils. Responses to hydrological conditions were less pronounced for Taxodium spp. If the results of this experiment are transferable to the field, then Acer rubrum, Fraxinus caroliniana, Pinus serotina, Taxodium ascendens, and Taxodium distichum seedlings can reasonably be expected to survive at least one year under a broad range of hydrological and edaphic conditions. With the exception of Taxodium spp., first-year growth for the species of this study can be facilitated by maintaining moist but not inundated conditions. These findings suggest that transfer of organic soils will benefit restoration and creation efforts, and that layering organic soil over mineral soil is more effective than using mineral soils or stockpiled topsoil.

Notes: Experimental plots of pulverized fuel ash (PFA), alone or mixed with flue gas desulphurization (FGD) gypsum, were seeded with topsoil from areas where PFA had been revegetated naturally, or with estuarine soil. Plots containing fresh PFA became more saline during the first three years due to formation of a salt crust, and these plots experienced colonization by halophytic species. Plots initially containing 2-year-old PFA declined in salinity throughout the experiment and were colonized by a richer plant community dominated by legumes. A total of 57 plant species was recorded in the two experiments. Of these, Melilotus officinalis (ribbed melilot), Medicago lupulina (black medick), Vulpia myuros (rat's tail fescue), and Puccinellia maritima (common saltmarsh grass) showed commercial potential for stabilizing these wastes, and by implication other saline or high-boron materials.

Notes: Limestone neutralization of the acidic water of Dog-way Fork in West Virginia resulted in significant improvements in both water quality and fish populations. Pretreatment water chemistry showed the stream to be highly acidified by acid precipitation, with pH under 5.0 and high aluminum concentrations. During treatment, the goals for the target area of pH 6.5 and acid-neutralizing capacity of 50 μmeq/L were met for 75.8% and 67% of the time, respectively. A pH 6.0 or above was maintained over 93% of the time. Monomeric aluminum concentrations were reduced significantly, and calcium to hydrogen ionic ratios were over 10 in the target area. Prior to treatment, no resident fish population was found in Dogway Fork. During five years of treatment, conditions were favorable for fish reproduction and survival. Eight fish species inhabited the stream, six were reproducing there, and a fishable brook trout population was established.

Notes: Duopolistic interaction between a small firm and a large established firm is considered and compared to guerrilla warfare, The paper investigates a “hit and run” equilibrium in which the small firm enters the market, stays there for several periods, exits, stays out for several periods, and then reenters. Occasionally there may be a price war (or retaliation), but the small firm may also exit voluntarily, thereby avoiding possible confrontation. The amount of time that the small firm stays in the market and the timing of the price wars do not follow any predictable pattern, which is part of the mixed strategies that both firms play in equilibrium.

Notes: This paper explores the potential of a digital camera to produce multiple images suitable for plotting. The objective was to build a three dimensional database by linking triangulated images from a Kodak DCS420 digital camera with a computer aided measurement system. The system then supplied X, Y, Z data from x, y image co-ordinates captured off two or more images. Relative accuracy in object space was around 1;50 000.

Notes: For centuries the ruins of Stonehenge have proved to be both a source of fascination and mystery for the many visitors to this World Heritage Site. The monument itself sets a puzzle that even to this day has not been solved, although there have been numerous theories suggested as to its construction. There is little accurate survey data currently available on the stones themselves, against which these many theories could be tested. The Photogrammetric Unit of English Heritage was therefore requested, in 1993, to commence a project to digitally map all the external faces of all the stones that form the visible remains of this internationally important monument. Photogrammetry was the chosen survey technique and this paper outlines the work of the Unit in using the latest digital photogrammetric equipment to generate a basic three dimensional model of Stonehenge.

Notes: The XVIIIth International Congress of Photogrammetry and Remote Sensing was held in the Austria Center, Vienna from 9th to 19th July, 1996. Reports on the Technical Commission Activities, on the Congress Exhibition and on the General Assembly were given at a meeting of the Photogrammetric Society on 7th October, 1996. Papers from the Congress were published in the International Archives of Photogrammetry and Remote Sensing, 31.

Notes: The paper deals with the determination of the metric quality of a 409634096 pixel high resolution digital camera back achieved from testfield measurements.

Notes: Digital photogrammetric techniques have opened fresh horizons and have enabled photogrammetry to become a more user friendly tool for a wide range of applications. However cost remains a critical issue, because new digital photogrammetric workstations and their related software are considered expensive. This paper examines the problem of digital rectification and presents low cost solutions involving the use of commercial off the shelf software packages, which include graphics and painting software, image processing tools and a CAD system. Additionally, specially developed software performing vector rectification is described and discussed. All the packages have been used for rectifying digital images and the results are presented. An attempt is also made to assess the accuracy and reliability of the results. Finally, several alternative digital products are presented, mainly concerning the use of the methods described in the restitution of cultural monuments.

Notes: Detailed understanding of the processes which control river bank erosion requires high resolution information concerning temporal changes in bank morphology. This paper describes the successful use of digital photogrammetry to extract high resolution digital elevation models (DEMs) from terrestrial oblique stereopairs of rapidly eroding river banks, using the commercial software package Erdas Imagine. This software was developed for use with aerial photography and satellite imagery; problems relating to the use of oblique terrestrial images are discussed and solutions presented. Photography was acquired using semi-metric cameras, mounted on tripods and positioned about 15m from the eroding bank. Data for DEM point spacings of 20mm were obtained, with accuracies of approximately ±12mm in depth. Digital photogrammetry can permit faster analysis, provide better accuracies and involve less ground disturbance than conventional methods of monitoring river channel change. Most importantly, DEM generation is considered to be more useful than traditionally acquired points or profiles for landform monitoring strategies.

Notes: This eighth and penultimate extract from the Archive is an unusual one for a number of reasons. Of these the most important one is the fact that it is concerned solely with instruments produced in Great Britain. It is therefore concerned mainly with the Thompson-Watts plotter, the only British instrument to go into full-scale commercial production. Strictly speaking the other instrument included in this extract should not have had a separate entry at all, simply because no instrument of this type exists now and so cannot have an entry in the catalogue of instruments. However, in order to trace the development of the Thompson-Watts Mk 2 plotter, produced in 1963, from the remarkable work of Henry Fourcade carried out in the 1920s, the Barr and Stroud ZA2 plotter based on his work has been accorded an Appendix of its own.Both instruments in this extract are based on an optical-mechanical solution to the photogrammetric problem of space intersection and the Thompson-Watts is in fact the only instrument of this type to be found in the catalogue. This particular solution did not find great favour on the continent and so few instruments of this type were produced commercially.

Notes: Internal eliminated segments (IESs) are sequences that interrupt coding and noncoding regions of germline (micronuclear) genes of ciliated protozoa. IESs are flanked by short, unique repeat sequences, which are presumably required for precise IES excision during macronuclear development. Coding and noncoding segments of genes separated by IESs are called macronuclear-destined segments, or MDSs. We have compiled the characteristics of 89 individual IESs in 12 micronuclear genes in the Oxytricha and Stylonychia genera to define the IES phenomenon precisely, a first step in determining the origin, function and significance of IESs. Although all 89 IESs among the 12 different genes are AT-rich, they show no other similarity in sequence, length, position or number. Two main types of IESs are present. IESs that separate scrambled MDSs are significantly shorter and more frequent and have longer flanking repeat sequences than IESs that intervene between nonscrambled MDSs. A comparison of the nonscrambled gene encoding β-telomere binding protein in three species of hypotrichs shows that even in the same gene IESs are not conserved in sequence, length, position, or number from species to species. A comparison of IESs in the scrambled gene encoding actin I in the three species shows that the evolutionary behavior of IESs in a scrambled gene may be more constrained. However, IESs in the scrambled actin I gene have shifted along the DNA molecule during evolution. In total, the various studies show that IESs are hypermutable in sequence and length. They insert, excise, and shift along DNA molecules more or less randomly during evolution, with no discernible function or consequences.

Notes: The phylogenetic relationships between major slime mould groups and the identification of their unicellular relatives has been a subject of controversy for many years. Traditionally, it has been assumed that two slime mould groups, the acrasids and the dictyostelids were related by virtue of their cellular slime mould habit; a view still endorsed by at least one current classification scheme, However, a decade ago, on the basis of detailed ultrastructural resemblances, it was proposed that acrasids of the family Acrasidae were not relatives of other slime moulds but instead related to a group of mostly free-living unicellular amoebae, the Schizopyrenida. The class Heterolobosea was created to contain these organisms and has since figured in many discussions of protist evolution. We sought to test the validity of Heterolobosea by characterizing homologs of the highly conserved glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from an acrasid, Acrasis rosea; a dictyostelid, Dictyostelium discoideum; and the schizopyrenid Naegleria andersoni. Phylogenetic analysis of these and other GAPDH sequences, using maximum parsimony, neighbour-joining distance and maximum likelihood methods strongly supports the Heterolobosea hypothesis and discredits the concept of a cellular slime mould grouping. Moreover, all of our analyses place Dictyostelium discoideum as a relatively recently originating lineage, most closely related to the Metazoa, similar to other recently published phylogenies of protein-coding genes. However, GAPDH phylogenies do not show robust branching orders for most of the relationships between major groups. We propose that several of the incongruencies observed between GAPDH and other molecular phylogenies are artifacts resulting from substitutional saturation of this enzyme.

Notes: We have developed a new staining procedure that combines the traditional Gram staining for bacteria and the Weber's chromotrope staining method, the standard technique for the detection of microsporidia spores in clinical Specimens. This “Gram-chromotrope” staining technique enhances the staining characteristics of microsporidia spores and facilitates the easy detection and differentiation of spores from other microorganisms that are found in clinical specimens, especially stool samples. This new technique is fast, reliable, and simple to perform, and can be easily adapted for use in clinical laboratories.

Notes: . The effect of conditioned media (media aspirated from a variety of cell cultures after 4 d of growth) on cellular invasion by sporozoites of the turkey coccidium, Eimeria adenoeides, was examined. Conditioned medium from turkey kidney cells and baby hamster kidney cells failed to alter invasion. However, conditioned medium from turkey cecal cell cultures produced a significant (P ≤ 0.05), two-fold increase in invasion over control medium in a variety of cell types. Retentates of conditioned medium from the turkey cecal cells that were passed through microconcentrators having molecular mass cutoffs of 50, 100, and 300 kDa similarly enhanced invasion over retentates from control medium. However, retentates from microconcentrators with a cutoff of 1,000 kDa failed to enhance invasion. Pretreatment in conditioned medium, followed by washing of sporozoites prior to inoculation into cultures, did not result in enhanced invasion. Moreover, when the interval between inoculation of sporozoites into cells and fixation of cultures was reduced to less than 3 h, no enhancement of invasion occurred. Conditioned medium from turkey cecal cells that were grown in the presence of 35S-translabel had at least two labeled bands at 150 kDa and 〉 200 kDa that were absent in conditioned media from turkey kidney and baby hamster kidney cells.

Notes: The virulence of Leishmania mexicana is determined by the concerted action of several parasite molecules. These cells lose their infectivity to host macrophages after prolonged cultivation in axenic growth media. Both virulent and attenuated variants of the parasite cells were cloned. The differential display reverse transcription-polymerase chain reaction technique was employed to understand whether this natural attenuation of the parasite cells is accompanied by differential expression of selected genes in those cells. Twelve different dinucleotide-anchored oligo(dT) antisense primers were used to make cDNAs from poly(A)+ mRNAs isolated from a clonal population of virulent and avirulent cells following a protocol optimized for Leishmania mRNAs. Those cDNAs were subjected to amplifications using each of the three different arbitrary decanucleotide primers and the corresponding anchored oligo(dT) primer. This procedure revealed four virulent-specific cDNA probes and one avirulent-specific cDNA probe. Differential expressions of these genes were confirmed by northern hybridization using the cloned cDNA probes. These results indicate that differential expression of genes may be the key in determining the molecular basis of leishmanial virulence.

Notes: . A unique group of entodiniomorph protozoa was found in forestomach contents from quokka (Setonix brachyurus), western grey kangaroo (Macropus fuliginosus), red kangaroo (Macropus rufus) and euro (Macropus robustus erubescens). A new genus, Macropodinium n.g., containing five new species, is described. Three species are described from forestomach contents of the quokka: Macropodinium baldense n. sp., Macropodinium moiri n. sp. and Macropodinium setonixum n. sp. A single species, Macropodinium ennuensis n. sp., is described from the red kangaroo and euro. The last species, Macropodinium yalanbense n. sp., is described in forestomach contents from the western grey kangaroo. At least three distinct features in the new genus are incompatible with any of the described families in the order Entodiniomorphida. On this basis, the new family Macropodiniidae has been created.

Notes: . Modifications of the arbitrarily primed polymerase chain reaction assay (i.e. a low annealing temperature and a very slow increase in the temperature during the elongation steps during the amplification cycles) allowed it to be used with the AT-rich Plasmodium falciparum DNA. The analysis of the products by polyacrylamide—urea gels, after silver staining, resulted in high resolution and sensitivity. Eighteen single and six combined pairs of arbitrary primers were tested. Two produced polymorphic patterns complex enough to differentiate between close Colombian isolates in a single assay. This method may be useful in studying the distribution and migration of strains in endemic areas, and for identifying intralaboratory cross-contamination of cultures.

Notes: . The 44-kDa regulatory subunit (R44) of one form of cAMP-dependent protein kinase of Paramecium was purified, and two partial internal amino acid sequences from it were used to clone the corresponding cDNA. This R44 cDNA clone was 1022-bp long, including 978 bp of coding sequence and 7 bp and 37 bp of 5′ and 3′ untranslated sequences, respectively. A 1.1-kb mRNA was labeled on a Northern blot. The deduced R44 amino acid sequence had 31%–38% positional identity to the sequences of other cloned cAMP-dependent protein kinase regulatory subunits. R44 sequence showed equal sequence similarity to mammalian types I and II regulatory subunits. The N-terminal sequence encoding the regulatory subunit dimerization domain found in most regulatory subunits is not present in the R44 clone, confirming the lack of regulatory subunit dimer formation previously reported for the Paramecium cAMP-dependent protein kinase. The putative autophosphorylation site of R44 contains the amino acid sequence TRTS, distinct from the consensus sequence RRXS, where X is any residue, found in other autophosphorylated cAMP-dependent protein kinase regulatory subunits and many cAMP-dependent protein kinase substrates.

Notes: We have constructed an arrayed, large insert, multiple coverage genomic library of Pneumocystis carinii DNA using the bacteriophage P1 cloning system. The library consists of ∽4800 independent clones with an average insert size of ∽55 kbp individually arrayed in 50 microtiter plates, and is readily screened on ten or fewer microtiter plate-sized filters using a high density colony replicating device. Screening of the library for unique P. carinii sequences detected an average of 4–5 positive clones for each, consistent with a several-fold coverage of the ∽10-mbp P. carinii genome. Restriction and hybridization analyses demonstrated that the P1 clones in this library are quite stable and contain few, if any, chimeric inserts. Thus, this arrayed, large insert library off. carinii genomic DNA will be a valuable tool in the future genetic dissection of this important pathogen.

Notes: The lipids of purified preparations of Pneumocystis carinii carinii freshly isolated from infected rats were analyzed and compared with those of whole lungs from normal and methylprednisolone-immunosuppressed uninfected rats. In this study, the neutral lipid fraction was examined in detail; the relative concentrations of individual classes making up this fraction were quantified. Of particular interest was the nature of the organism's ubiquinone (coenzyme Q, CoQ) fraction because atovaquone, a hydroxynaphtho-quinone (566C80) analog of ubiquinone, is efficacious in the treatment of P. carinii pneumonia. The ubiquinone concentration in both P. carinii and lung tissues was relatively low compared to that present in rat heart and liver tissues. Two homologs were identified in the organism: CoQ10 was the predominant homolog with lesser amounts of CoQ9 present. In contrast, the lungs of normal and immunosuppressed uninfected rats had CoQ9 and lesser amounts of CoQ8, but no detectable CoQ10. Furthermore, radiolabeled mevalonic acid was incorporated in vitro into the ubiquinone fraction of P. carinii indicating that the organism has the de novo branch of the isoprenoid biosynthetic pathway leading to polyprenyl formation. Hence, it was concluded that CoQ10 (if not both CoQ110 and CoQ9) in P. carinii as not scavenged from the host but was synthesized by the organism. Although lung tissues contained substantial free fatty acids, the organism was enriched in these lipids. The high concentration of free fatty acids and relatively low level of triglycerides in P. carinii suggest that fatty acids may represent major carbon sources for ATP production by the organism.

Notes: . Paulinella indentata n. sp. is described from benthic sandy sediments of Kames Bay, Scotland. The description is based on light and scanning electron microscopical observations. This marine, testate amoeba has filose pseudopodia, sometimes branched, extending up to 50 μm from the aperture. Tests are oval in outline (c. 15.8 × 9.8 μm) pale yellow in colour with a short collar. The most important diagnostic feature is the morphology of the surface scales which are arranged in staggered rows. Each cell is covered with around 22 scales (c. 5.7 × 2.9 μm). Scales are rectangular, curved with rounded corners and markedly indented along their median axis. They have a hollow channel running below each ridge and their surface is punctuated with rows of pores. This is the first isolation of Paulinella from benthic marine sediments.

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

Notes: Until recently, Tetrahymena thermophila has rarely been isolated from nature. With improved sampling procedures, T. thermophila has been found in ponds in many northeastern states. The availability of resident populations makes possible both population and ecological genetic studies. All seven known mating types have been recovered; no eighth mating type has been found. Crosses among whole-genome homozygotes derived from Pennsylvania isolates reveal a spectrum genotypes with mating type alleles resembling traditional A (IV- and VII-) and B(I-) categories. The genotypes differ significantly with respect to mating type frequency, both among themselves and from previously described genotypes. One A-category genotype appears to lack mating type II, while one A-category and all B-category genotypes have low frequencies of mating type III, thus accounting for the low frequency of III in the pond. The low frequency of III in all five B-category genotypes examined suggests that the founding allele in this region was low for III. These and other differences are discussed both in terms of mating type frequencies in the pond and in terms of the possible molecular structure of mat alleles. By contrast, numerous variants of the cell surface immobilization antigen are found in addition to the previously described i-antigens. Variants of the known SerH alleles include those with restriction fragment length polymorphisms and temperature sensitivity as well as alleles with new antigenic specificity. Multiple alleles are present in single ponds. Genes exhibiting serially dominant epistasis over SerH genes also are found. In two instances (K and C), families of antigenically similar polypeptides are expressed in place of H i-antigen. Molecular weight differences suggest that these paralogous i-antigen genes evolve by gene duplication and unequal crossing over within central repeats. The existence of complex patterns of epistasis together with seasonal changes in i-ag frequencies suggest that i-ag play an important, but as yet unknown, ecological role related to the occurrence of frequent conjugation.

Notes: DNA polymorphisms of different strains of Blastocystis isolated from humans, a chicken, and a reptile were examined by an arbitrary primer PCR method. Two strains of Blastocystis hominis isolated from humans in the USA and Japan yielded nearly identical PCR products. However, one strain of B. hominis (isolated from a human in Singapore) yielded quite different PCR products. Blastocystis sp. isolated from a chicken yielded PCR products similar to those of the former two strains, while Blastocystis lapemi, isolated from a reptile, shared no bands with any of the other isolates. These results indicate the possibility that our isolate from the chicken is a zoonotic strain, and that there is intraspecific variation of Blastocystis hominis.

Notes: In searching the genomes of early-diverging protists to study whether the possession of calmodulin is ancestral to all eukaryotes, the gene for calmodulin was identified in Trichomonas vaginalis. This flagellate is a member of the Parabasalia, one of the earliest lineages of recognized eukaryotes to have diverged. This sequence was used to isolate a homologous 1.250-kb fragment from the T. vaginalis genome by inverse polymerase chain reaction. This fragment was also completely sequenced and shown to contain the 3′ end of the single-copy calmodulin gene and the 3′ end of a gene encoding a protein with high similarity to E2 ubiquitin-conjugating enzymes, a family which has previously only been identified in animals, plants, and fungi. Phylogenetic analysis of 50 members of the E2 family distinguishes at least nine separate subfamilies one of which includes the T. vaginalis E2-homologue and an uncharacterized gene from yeast chromosome XII.

Notes: Classification of Acanthamoeba at the subgenus level has been problematic, but increasing reports of Acanthamoeba as an opportunistic human pathogen have generated an interest in finding a more consistent basis for classification. Thus, we are developing a classification scheme based on RNA gene sequences. This first report is based on analysis of complete sequences of nuclear small ribosomal subunit RNA genes (Rns) from 18 strains. Sequence variation was localized in 12 highly variable regions. Four distinct sequence types were identified based on parsimony and distance analyses. Three were obtained from single strains: Type T1 from Acanthamoeba castellanii V006, T2 from Acanthamoeba palestinensis Reich, and T3 from Acanthamoeba griffini S-7. T4, the fourth sequence type, included 15 isolates classified as A. castellanii, Acanthamoeba polyphaga, Acanthamoeba rhysodes, or Acanthamoeba sp., and included all 10 Acanthamoeba keratitis isolates. Interstrain sequence differences within T4 were 0%–4.3%, whereas differences among sequence types were 6%–12%. Branching orders obtained by parsimony and distance analyses were inconsistent with the current classification of T4 strains and provided further evidence of a need to reevaluate criteria for classification in this genus. Based on this report and others in preparation, we propose that Rns sequence types provide the consistent quantititive basis for classification that is needed.

Notes: Nucleoside diphosphate kinase (NDP kinase) from Paramecium was purified to homogeneity. The native enzyme was 80 kDa (by gel filtration), with subunits of 18 and 20 kDa. Near the amino terminus, 15 of 20 residues were identical with those in human NDP kinase, and 17 of 20 with the awd gene product from Drosophila. NDP kinase bound α-labeled ATP and GTP, and a photoreactive GTP analog labeled both subunits. Purified NDP kinase underwent autophosphorylation on a histidine and a serine residue using either ATP or GTP as a substrate. The enzyme also catalyzed acid-stable phosphorylation of casein and phosvitin. This protein kinase activity is distinct from the histidine phosphorylation that is part of the NDP kinase catalytic cycle. Antiserum against the purified protein from Paramecium cross-reacted with 16- to 20-kDa proteins in most species tested, and with a larger protein (44 kDa) in Paramecium, Xenopus, and two human lines. The multiple forms (20 and 44 kDa) of the NDP kinase in Paramecium and its protein kinase activity, suggest that the protein is more than a housekeeping enzyme; it may have regulatory roles such as those of the NDP kinase-like awd protein of Drosophila and Nm23 protein of humans.