Studies on plating efficiency and estimation of viability of suspensions of Paracoccidioides brasiliensis yeast cells

Mild sonication was used to obtain single cell suspensions of Paracoccidioides brasiliensis. These cells were intact by microscopic criteria. Direct cell counts in a given inoculum and colony formation on various media were used to determine plating efficiency. Sonicated and nonsonicated cell suspensions were used to study plating efficiency and to estimate viability by means of vital dyes. Methylene blue, Erythrosin B, and Janus green were unreliable when used with P. brasiliensis, but vital dyes were accurate when tested with Candida albicans.Acridine orange gave more meaningful results of viability. Estimates of viability, however, changed significantly as a result of relatively minor alterations in the composition of the suspending medium.In initial experiments, the plating efficiency of P. brasiliensis was dismally low. It descended abruptly with increasing dilution of inoculum. Efficiency was much improved if horse serum was added to brain heart infusion plates or if glucose glycine yeast extract (GGY) plates were incubated at room temperature and mycelial colonies were counted. With the technique we report, current plating efficiency of sonicated suspensions is of the order of 25 %. Our results and procedures have an important bearing upon those studies concerned with in vitro killing of P. brasiliensis in suspensions or with isolating this fungus from clinical or environmental specimens.     

Cellular immunity to Coccidioides immitis: In vitro lymphocyte response to spherules,arthrospores, and endospores

In vitro stimulation of incorporation of tritiated thymidine by human peripheral lymphocytes in response to two soluble antigens and three different intact but nonviable fungal forms of Coccidioides immitis was studied. Lymphocytes were obtained from three groups of subjects: healthy skin test positive, healthy skin test negative, and disseminated disease. Dose-response relationships to the intact forms (endospores, arthrospores, and spherules) were determined. Responses of lymphocytes from healthy skin test-positive subjects and subjects with disseminated disease were similar. Ranking of antigens by “potency” gave the following results: endospores = spherulin > mycelial filtrate > arthrospores = spherules. Endospores were the most potent of the intact forms in 10 of 11 subjects. The clear superiority of endospores over spherules is not due to differences in the total particle surface area available for presentation to the leukocytes. All antigens tested except spherules could discriminate between skin test-positive and skin test-negative subjects in this in vitro system. A T-cell-enriched, B-cell- and mono-cyte-depleted cell population demonstrated an active response to spherulin and to endospores. The variance of these finding with animal studies demonstrating spherules to be immunogenically superior when compared to endospores is discussed. This may have importance in future studies in humans of vaccines to C. immitis.     

Development of a radiometric metabolic viability testing method for human and porcine skin

A radiometric viability assay based upon the conversion of [¹⁴C]glucose into ¹⁴CO₂ by the viable cells on the dermal side of whole skin has been developed. The assay proved to be sensitive, reproducible, and practical, and was based upon the use of a microbiological growth detection system commonly used in many hospitals and laboratories. Relatively small samples of skin (0.25–1.00 g) were used in the test, and it was found that microbiological contamination did not interfere with the assay under normal conditions. The linear proportionality of the assay with both time and amount of skin assayed precluded the difficulties of nonlinear proportionality in other systems, allowing direct comparisons to be made between skin samples of different sizes and different incubation times. The assay could also detect ¹⁴CO₂ released from many radiolabeled substrates, including glucose, aspartate, glutamate, ornithine, orotic acid, and glycerol. Thus, the method could be used to test a number of cellular functions necessary for viability, including glycolysis, the functioning of the citric acid cycle and the pentose phosphate pathway, sugar and amino acid metabolism, pyrimidine biosynthesis, and cryopreservative agent metabolism. Since any of these tests could be performed in 4 hr, a viability assay based upon glycolysis alone, or in combination with any of the other tested substrates, could be carried out after allograft skin procurement before a decision needed to be made on skin cryopreservation.     

Inland salt waters of southern Africa

Inland salt lakes are widely distributed in southern Africa: they are particularly common in South Africa, but many occur in Namibia and Botswana. All are shallow, and most are ephemeral with salinities that are not very high (mostly < 50 g l⁻¹). Fringing zones of halophytes or submerged macrophytes are neither well-developed nor taxonomically diverse. The Cyanobacteria, especially Nodularia spumigena, often dominate the phytoplankton. The fauna of the Makgadikgadi area (northeast Botswana) is diverse and is similar to that of East African salt lakes. The aquatic fauna of salt water south of the Makgadikgadi Basin, on the other hand, is extremely depauperate, has no well-defined assemblage confined to saline waters, and appears mostly to comprise tolerant freshwater forms. Lovenula falcifera and Metadiaptomus transvaalensis (diaptomid copepods), Moina micrura (Cladocera) and Brachionus plicatilis (Rotifera) are frequently encountered zooplankton species, a few species of insects (Anisops sp., beetles, chironomids and ephydrids) are the principal non-planktonic macroinvertebrates. Artemia ‘salina’ is occasionally present, but may be an introduced form. The avifauna, in contrast to the aquatic macroinvertebrate fauna, is rich, with the greater and lesser flamingo often common.     

Amphipod crustaceans as an important component of zoobenthos of the shallow Antarctic sublittoral

Benthic quantitative samples were taken in 1988 in the soft bottom sublittoral of Admiralty Bay (King George Island, South Shetlands) using a Tvärminne-type bottom sampler and SCUBA-diving technique at 7 successive stations situated at depths from 4 to 30 m.Dominant animal groups in terms of abundance were Amphipoda, Polychaeta and Bivalvia, whereas in terms of biomass Echinoidea were also dominant. Amphipod crustaceans clearly dominated the zoobenthos at depths from 10 to 25 m (the numerical share surpaising 60%) with maximal abundance of abt. 17 000 ind m^–2; in terms of biomass at specific depths amphipods occupied the 1st, 2nd or 3rd place with maximal biomass of abt. 100 g m^–2 where the maximal total biomass of zoobenthos reached 260 g m^–2 (10 m).Amphipoda were the most diversified group with some 35 taxa belonging to 14 families. Most species belonged to Eusiridae s.l. and Lysianassidae s.l. Dominant forms were Pontogeneiella brevicornis, Prostebbingia gracilis, Schraderia gracilis, Hippomedon kergueleni, Orchomenella cf. ultima, Cardenio paurodactylus and Paraphoxus rotundifrons.     

Disturbance regimes as determinants of seed banks in coastal dune vegetation of the southeastern Cape

Soil-stored seed banks of grassland, fynbos and thicket, all growing on calcareous dunes and each subject to different disturbance regimes, were examined. Seed banks were determined from counts of germinants from 50 soil cores from each type. Aboveground estimates of plant species cover in 10 1-m² plots were used in determining vegetation/seed bank similarities. There was no evidence for seed bank densities to be markedly higher in the most frequently disturbed community (grassland -4273 seeds/m²) than the least disturbed community (thicket - 3417 seeds/m²). Highest similarity between seed bank and above-ground vegetation composition in terms of species and growth form/life-span classes was recorded for grassland (CC = 50%). Lowest similarity (CC = 13%) was found in the less frequently disturbed thicket where no seeds of climax trees were recorded in the seed bank. A fynbos community on a north-facing (warm, dry) slope had intermediate-sized seed banks (1683 seeds/m²) with intermediate vegetation/seed bank similarity (CC = 46%). However, on the south-facing slope, which has a large post-fire ephemeral herb component, seed banks were larger (4518 seeds/m²) but less similar to above-ground vegetation (CC = 39%o). Ordination (DCA) of vegetation data from the four communities was different from an ordination of their seed bank data. Fynbos shrub species were absent from seed banks of both grassland and thicket, even though secondary succession proceeds from grassland, through fynbos to thicket. Their seed banks appear less persistent than those of European heath or Californian chaparral shrubs.     

Yearly maps of summer temperatures in Western Europe from A.D. 1750 to 1975 and Western North America from 1600 to 1982:

The primary purpose of this publication is to make available two series of historical maps showing summer (April–September mean) temperatures over western Europe from A.D. 1750 and western North America from A.D. 1600. These maps have been reconstructed using networks of temperature-sensitive tree-ring chronologies as part of a continuing collaboration between the Swiss Federal Institute for Forest, Snow and Landscape Research (FSL) at Birmensdorf, Switzerland and the Climatic Research Unit (CRU) at the University of East Anglia, Norwich, U.K. The maps were reconstructed using relatively long maximum latewood density chronologies selected from the extensive networks of tree-ring data produced at the FSL. The reconstructions were produced using a spatial regression technique developed at CRU.The basis for the reconstructions are wood samples from various conifers growing at cool-moist sites close to the alpine or boreal timberlines. This material was analysed by x-ray densitometry and the resulting maximum latewood density chronologies were calibrated against meteorological data over a 100-year calibration period.These results emphasize the value of dendroclimatological research which can provide continentalscale reconstructions of past climate which represent a potential resource for climatologists, biologists, and historians with an interest in the spatial variability of summer temperatures over the middle to high latitudes.     

Altitudinal replacement of Ephemeroptera in a subtropical river

An altitudinal sampling transect was established in a subtropical mountain river to determine if replacement between two mayfly faunal components (oligostenothermal and polystenothermal) occurred. We detected three main groups: Group A, species with high abundance at higher stations, declining abruptly downstream; Group B, species with maximum abundance at lower elevations; either declining abruptly upstream or absent from higher elevations; and Group C, species with little elevational change. The physical and chemical variables did not change abruptly with altitude during our winter sampling, although the distribution of groups A and B species did. The observed distributional patterns of groups A and B do not appear to be correlated with variations in the physical and chemical variables surveyed.     

Two new species of Viola (Violaceae) from the Intermountain West,U.S.A.

Viola lithion andV. franksmithii are rock-dwelling endemics that belong to the North American perennial, caulescent, simple-leaved, blue-, purple-, or white-flowered group of species.Viola lithion, from the White Pine and Pilot mountain ranges of eastern Nevada and adjacent Utah, belongs to sectionChamaemelanium subsectionCanadenses. It most closely resembles the reniform-leaved, Olympic MountainV. flettii, together with which it possibly shares an ancestry with the widespread and allopatricV. canadensis. Viola franksmithii, restricted to the Logan Canyon drainage of the Bear River Range in northern Utah, belongs to sectionRostellatae subsectionRosulantes. It comes closest morphologically toV. howellii of the Coast Range, but likely evolved from the widespreadV. adunca or a common ancestor.     

Freshwater sponges (Porifera: Spongillidae) from Panamá

Aquatic habitats in the Republic of Panamá were examined for freshwater sponges during 21 Feb. to 6 March 1986. Eunapius carteri, Trochospongilla leidyi, and Trochospongilla horrida were found in the northern canal area. This is the first report of E. carteri and T. horrida from Central America, and the first record of E. carteri from the Western Hemisphere. Taxonomic characters used to distinguish T. leidyi and T. horrida from similar South American species are discussed.