MEMBRANE LIFTING TECHNIQUE FOR RAPID DETECTION OF ESCHERICHIA COLI 0157:H7 IN INOCULATED GROUND MEATS

A membrane lifting technique was developed for direct rapid detection of Escherichia coli 0157:H7 in inoculated ground meats. Duplicate groups of 2 meatballs were inoculated with volumes of 0.1-ml of a serial dilution (1:10) of E. coli 0157:H7 or a mixed culture containing one strain of E. coli 0157:H7 and a non-0157:H7E. coli serotype (E. coli ATCC 25922). Each meatball was sampled by sandwiching with 2 pieces of nitrocellulose membranes and pressing against each other to the center of the meatball. The membranes were in contact with the meats for 10 min to lift the bacteria. The membranes were removed and incubated on MacConkey-sorbitol agar plates with the meat contact side up. After 18 h incubation at 37C, an immunostain was performed directly on the membranes for detection of the presence or absence of E. coli 0157:H7. This method was found to be sensitive enough to detect as few as 1 to 2 cells of E. coli 0157:H7 inoculated on surfaces of 18-g meatballs. This method might be used as a rapid presumptive test for E. coli 0157:H7.     

Biofilm development and invertebrate colonization of wood in four New Zealand streams of contrasting pH

1. Biofilm development and activity on wood substrata (Nothofagus menziesii) were examined at four forested sites in a South Island, New Zealand, river catchment over a period of 6 months. Two of the sites had brown waters and mean pH of 3.7 and 4.5, whereas the other two had clear waters and mean pH of 6.3 and 6.8. 2. Fungi and other filamentous heterotrophs were the dominant colonizers of wood at all sites; few algal cells were present. Incorporation of ¹⁴C-glucose by biofilms was greatest in all four streams after 3 months, whereas endocellulase activity fluctuated over time and temporal patterns differed among streams. 3. No clear relationship was found between the incorporation of ¹⁴C-glucose or endocellulase activity of biofilms and pH, although at one near-neutral pH site ¹⁴C-glucose uptake increased in response to nutrient (N + P) additions. 4. After 6 months, incorporation of ¹⁴C-glucose and endocellulase activity of biofilms on Pinus radiata dowels buried vertically in the stream beds did not differ at depths of 3–9 cm and 19–25 cm in each stream. 5. Radiotracer experiments with a grazing amphipod (Paraleptamphopus sp.) demonstrated that biofilms on wood from all four sites could be ingested and at least partially assimilated. Chironomid larvae and harpacricoid copepods were the most abundant invertebrates colonizing wood substrata at all sites. Different chironomid species dominated at acidic and near-neutral pH sites. 6. Overall, our findings provide little support for the hypothesis that microbial activity on organic substrata is necessarily lower in streams of low pH.     

The encrustation of fossil and recent sea-urchin tests: ecological and taphonomic significance

A comparison of fossil ( Echitwlampas sp. from the Lower Miocene Zogelsdorf Formation, Austria) and Recent ( Schizaster canaliferus from the northern Adriatic Sea) irregular sea-urchin tests shows that, upon their death, burrowing echinoids can serve as a substrate for a dense epigrowth. Size, shape, stable orientation, and surface residence-time were identified as key factors governing encrustation. The encrusters on fossil Echinolampas were bryozoans, serpulid polychaetes, barnacles, and coralline algae, while the recent material was initially encrusted by serpulids and hydrozoan colonies, and ultimately covered by the full range of sessile, hemi-sessile, and vagile species characterizing the benthic community in the Adriatic. In Echinolampas , epigrowth was more abundant on the lower (oral) surface; this specific distribution was echoed in S. canaliferus , where epigrowth started on the bottom side and grew upward. This indicates that the tests have a stable orientation and a surface residence-time long enough to allow intense encrustation. A taphonomic model is developed, and the role of encrustation on such special substrates for overall community structure is discussed. The Recent/fossil comparison provides new insights for both fields of study: the recent material indicates the role of soft-bodied faunas as well as the complexity of small-scaled ecological processes; the fossil material reflects many of the above phenomena and adds important taphonomic details on the fate of encrusted biogenic structures and on encruster growth patterns and distributions.     

Seasonal changes of osmotic pressure, symplasmic water content and tissue elasticity in the blades of dune grasses growing in situ along the coast of Oregon

Abstract Midday water potentials of blades of the dune grasses Ammophila arenaria (L.) Link and Elymus mollis Trin. ex Spreng. growing in situ declined over the summer growing period, indicating a trend of increasing water stress. An analysis of the water relations characteristics of these blades using pressure-volume techniques demonstrated that both species increased bulk osmotic pressure at full hydration () and, therefore, bulk turgor as an acclimation response. In A. arenaria, however, the increase of osmotic pressure (+ 0.35 MPa) was entirely the result of decreasing symplasmic water content. The increase of osmotic pressure (+ 0.54 MPa) observed in E. mollis blades was due to solute accumulation (72% of Δ) and to a lesser degree, decreased symplasmic water content (28% of Δ). Osmotic adjustment in E. mollis blades was accompanied by a significant decrease in tissue elasticity (_max went from 12 to 19 MPa). The elastic properties of A. arenaria blades remained constant over the same period and had a maximum modulus (10 MPa) that was always less than that of E. mollis, As estimated from Höfler plots, these seasonal adjustments of osmotic pressure and differences in tissue elasticity enabled plants in situ to maintain turgor pressure in the range of 0.5–0.6 MPa at the lowest water potentials of mid-August. Laboratorygrown plants exhibited the species-specific differences in osmotic pressure, turgor pressure, and tissue elasticity observed in field plants. Although certain alterations of leaf structure were expected to coincide with the observed changes and species-specific differences in symplasmic water content and tissue elasticity, these could not be detected by measurements of specific leaf weight or the ratio of dry matter to saturated water content.     

Calcification in aquatic plants

Abstract. The CaCO₃ deposits of aquatic plants may be intra-, inter- and extracellular. Calcification is mainly the result of photosynthetic CO₂ or HCO⁻₃ assimilation. This raises the local pH and CO²⁻₃ concentration resulting from shifts in the dissolved inorganic carbon equilibrium, due to either net CO₂ depletion as in Halimeda or localized OH⁻ efflux (or H⁺ influx) as in Chara. The plant cell wall may be important in CaCO₃ nucleation by acting as an epitaxial substratum or template, or by creating a microenvironment enriched in Ca²⁺ compared to Mg²⁺. Hypotheses on the reason for the lack of calcification in many aquatic plants are presented.     

The Possible Involvement of Cyclic AMP and Volatile Substance (s) in the Development of a Macrocyst-Forming Strain of Dictyostelium mucoroides

A mutant MF1 previously isolated from Dictyostelium mucoroides -7 (Dm7) formed macrocysts with or without light when plated on agar at high cell dinsities. At lower cell densities, however, the MF1 cells formed only fruiting bodies. This failure to form macrocysts was shown to be due to the subthreshfold concentration of a volatile substance(s) required for macrocyst formation. Although ammonia is a volatile substance produced by both the Dm7 and MF1 cells, no evidence of its involvement in macrocyst formation was obtained. Mixing the Dm7 and MF1 in a one-to-one ratio resulted only in fruiting body formation suggesting that the Dm7 cells produced a factor which allowed MF1 cells to form fruiting bodies. This factor may be cyclic AMP (cAMP) since addition of cAMP to the medium directed development of MF1 cells to fruiting body formation. The effect of cAMP was exhibited most conspicuously when MF1 cells were exposed at the aggregation stage. Based on these results it is suggested that developmental pathway of the D. mucoroides macrocystforming strain Dm7 and its mutant MF1 may be determined by the relative concentrations of the volatile, macrocyst-inducing substance(s) and cAMP at the aggregation stage.     

Algal borings and framboidal pyrite in Upper Ordovician brachiopods

Algal borings in the shells of the articulate brachiopods Plaesiomys subquadrata (Hall) and Hebertella sinuata (Hall) from the Richmond Formation of Ohio are empty, or partially to entirely filled with pyrite. The pyrite occurs as single framboids and other crystal forms, or in chains filling the bores. The borings provide some insight into the early diagenetic history of the Richmond sediments near Cincinnati. Pyritization probably occurred within a few years, only a short distance (a few centimeters) below the sediment surface through the activities of sulfur-reducing bacteria. Pyrite precipitated around a nucleus such as a bacterium or algal cell, or developed within an organic structure such as an algal cell or organic membrane.     

Why change mates?

In some wading birds (Charadrii) each adult rears a brood alone. The female leaves her first clutch with her first mate before herself rearing a clutch fertilized by a second male, who is already tending its first mate's first clutch. We here develop a simple model to account for the exchange of mates between clutches, and relate it to reported field studies. We suggest that females are attempting to be polygamous, rather than simply bigamous. Provided a female has a chance of obtaining a second, unmated male and that the costs of leaving the first male before remating are not high, matechanging will be favoured over monogamy. The implications of this model to the evolution of the more usual forms of polyandry and of male-like females are discussed.     

Distribution of Fibronectin and Collagen during Mouse Limb and Palate Development

Indirect immunofluorescence has been used to study the distribution of fibronectin and collagen types I, II, and III in the developing primary and secondary palatal processes and forelimb buds of the Swiss Webster (NIH) mouse. In the palatal processes fibronectin and types I and III collagen are distributed throughout the mesenchyme. Fibronectin is present in the basement membrane, while types I and III collagen are localized in a linear, discontinuous fashion beneath the basement membrane. Fibronectin is not observed in the epithelium, including the presumptive fusion areas. In the forelimb bud these components show a similar distribution prior to chondrogenesis (early day 11). When chondrogenesis commences (late day 11 or early day 12) fibronectin and, to a lesser degree, types I and III collagen are apparently concentrated in the core mesenchyme, suggesting that fibronectin has a role in initiating chondrogenesis, perhaps by increasing cellular aggregation. Type II collagen is observed only in chondrogenic regions. The codistribution of fibronectin and types I and III collagen supports in vitro studies which indicate that cells use fibronectin to bind to collagen in the matrix. The developing chondrogenic regions appear to lose fibronectin gradually, concomitant with the appearance of type II collagen, suggesting that fibronectin is not involved in the maintenance of functional chondrocytes in their matrices.