Cumulative sedimentation analysis of Escherichia coli debris size

A new method to measure Escherichia coil cell debris size after homogenization is presented. It is based on cumulative sedimentation analysis under centrifugal force, coupled with Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis of sedimented proteins. The effects that fermentation and homogenization conditions have on the resulting debris distributions were investigated using this method. Median debris size decreased significantly from approximately 0.5 mu m to 0.3 mu m as the number of homogenization passes increased from 2 to 10. Under identical homogenization conditions, uninduced host cells in stationary phase had a larger debris size than exponential cells after 5 homogenizer passes. This difference was not evident after 2 or in passes, possibly because of confounding intact cells and the existence of a minimum debris size for the conditions investigated. Recombinant cells containing protein inclusion bodies had the smallest debris size following homogenization. The method was also used to measure the size distribution of inclusion bodies. This result compared extremely well with an independent determination using centrifugal disc photosedimentation (CDS), thus validating the method. This is the first method that provides accurate size distributions of E. coli debris without the need for sample pretreatment, theoretical approximations (e.g. extinction coefficients), or the separation of debris and inclusion bodies prior to analysis. (C) 1997 John Wiley & Sons, Inc.

Holocene sediments of Wistari Reef: towards a global quantification of coral reef related neritic sedimentation in the Holocene

Wistari Reef. within the southern Great Barrier Reef. is a shallow coral reef platform featuring a very clearly defined leeward accretionary wedge of carbonate sediments. The total global area of shallowly submerged coral reef has been quantified as 255 000 km(2). The question then becomes: What additional area of sediment of significant thickness is associated with the measured shallow reef areas T At Wistari Reef, the leeward sedimentary wedge has an area and a thickness that are roughly equal to the Holocene sediments that have accumulated on the platform. Several important observations can be made from these data. Firstly. the area of significant neritic carbonate sedimentation ( > 1 m/ka) associated with coral reefs is near 500000 km(2). Secondly, the production rate of neritic carbonates at Wistari Reef is almost 50%, less than the accumulation rate needed to obtain the volume of Holocene reef sediments observed. This implies that both production and accumulation neritic carbonate must have been more than a factor of two higher in the early to mid Holocene. (C) 2001 Elsevier Science B.V. All rights reserved.

Studies of solute self-association by sedimentation equilibrium: allowance for effects of thermodynamic non-ideality beyond the consequences of nearest-neighbor interactions

A sedimentation equilibrium study of a-chymotrypsin self-association in acetate-chloride buffer, pH 4.1 I 0.05, has been used to illustrate determination of a dimerization constant under conditions where thermodynamic non-ideality is manifested beyond the consequences of nearest-neighbor interactions. Because the expressions for the experimentally determinable interaction parameters comprise a mixture of equilibrium constant and excluded volume terms, the assignment of reasonable magnitudes to the relevant virial coefficients describing non-associative cluster formation is essential for the evaluation of a reliable estimate of the dimerization constant. Determination of these excluded volume parameters by numerical integration over the potential-of-mean-force is shown to be preferable to their calculation by approximate analytical solutions of the integral for this relatively small enzyme monomer with high net charge (+ 10) under conditions of low ionic strength (0.05 M). (C) 2001 Elsevier Science B.V. All rights reserved.

Integrated treatment of shrimp effluent by sedimentation, oyster filtration and macroalgal absorption: a laboratory scale study

Effluent water from shrimp ponds typically contains elevated concentrations of dissolved nutrients and suspended particulates compared to influent water. Attempts to improve effluent water quality using filter feeding bivalves and macroalgae to reduce nutrients have previously been hampered by the high concentration of clay particles typically found in untreated pond effluent. These particles inhibit feeding in bivalves and reduce photosynthesis in macroalgae by increasing effluent turbidity. In a small-scale laboratory study, the effectiveness of a three-stage effluent treatment system was investigated. In the first stage, reduction in particle concentration occurred through natural sedimentation. In the second stage, filtration by the Sydney rock oyster, Saccostrea commercialis (Iredale and Roughley), further reduced the concentration of suspended particulates, including inorganic particles, phytoplankton, bacteria, and their associated nutrients. In the final stage, the macroalga, Gracilaria edulis (Gmelin) Silva, absorbed dissolved nutrients. Pond effluent was collected from a commercial shrimp farm, taken to an indoor culture facility and was left to settle for 24 h. Subsamples of water were then transferred into laboratory tanks stocked with oysters and maintained for 24 h, and then transferred to tanks containing macroalgae for another 24 h. Total suspended solid (TSS), chlorophyll a, total nitrogen (N), total phosphorus (P), NH4+, NO3-, and PO43-, and bacterial numbers were compared before and after each treatment at: 0 h (initial); 24 h (after sedimentation); 48 h (after oyster filtration); 72 h (after macroalgal absorption). The combined effect of the sequential treatments resulted in significant reductions in the concentrations of all parameters measured. High rates of nutrient regeneration were observed in the control tanks, which did not contain oysters or macroalgae. Conversely, significant reductions in nutrients and suspended particulates after sedimentation and biological treatment were observed. Overall, improvements in water quality (final percentage of the initial concentration) were as follows: TSS (12%); total N (28%); total P (14%); NH4+ (76%); NO3- (30%); PO43-(35%); bacteria (30%); and chlorophyll a (0.7%). Despite the probability of considerable differences in sedimentation, filtration and nutrient uptake rates when scaled to farm size, these results demonstrate that integrated treatment has the potential to significantly improve water quality of shrimp farm effluent. (C) 2001 Elsevier Science B.V. All rights reserved.

Exact theory of sedimentation equilibrium made useful

The exact description of the thermodynamics of solutions has been used to describe, without approximation, the distribution of all the components of an incompressible solution in a centrifuge cell at sedimentation equilibrium. Thermodynamic parameters describing the interactions between solute components of known molar mass can be obtained by direct analysis of the experimental data. Interpretation of the measured thermodynamic parameters in terms of molecular interactions requires that an arbitrary distinction be made between nonassociative forces, like hard-sphere volume-exclusion and mean-field electrostatic repulsion or attraction, and specific short-range forces of association that give rise to the formation of molecular aggregates. Provided the former can be accounted for adequately, the effects of the latter can be elucidated in the form of good estimates of the equilibrium constants for the reactions of aggregation.

Sedimentation pelagique et encroûtements cryptalgaires: les calcaires gremeleux du Carixien portugais

In Portugal, Carixian is generally represented by alternative layers of marly limestones characterized by nodule and lumpy levels. These layers are particularly developped [show preferential development] on passage areas to a sedimentary basin, particularly along the slope of tilted blocks between the Meseta and Berlenga's horst. This facies is included in the range of the «nodular limestone» and of the «ammonitico-rosso». Limestones are radiolaria micrites with fragments of pelagic organisms (ammonoids, thin shelled gastropods). These layers can be affected by intensive bioturbation (Brenha) which is responsible for dismantlement, specially where the initial thickness does not exceed a few centimetres. This process can lead to the isolation of residual nodules (Brenha, São Pedro de Muel, Peniche) which can be mobilised by massive sliding (Peniche). The isolated elements, shell fragments or residual nodules, can also be incrustated, thus developing oncolitic cryptalgal structures. At Brenha the lump structure developed progressively into a sequence overlapping the normal sedimentary one (thick limestone beds alternating with bituminous shales). Cryptalgal structures correspond to rather unstable environment conditions on mobile margins. These structures are known in deep pelagic sediments corresponding to well defined events of the geodynamic evolution (end of the initial rifting). Cryptalgal accretions disappear towards the sedimentary basin, and the nodular levels are less important. In the articulation areas with the Tomar platform, small mounds and cupules (Alcabideque) developed within the alternating marly-limestone levels. They represent the so called «mud mounds» of metric dimensions. The upper part of these «mud mounds» is hardened, showing track remains and supporting some brachiopods and pectinids. Hence the lumpy facies of Portugal is included among the range of sedimentaty environments and can be used as «geodynamic tracer».

Le gisement de Asseiceira, Portugal (Miocène supérieur) Données complémentaires sur Freiria de Rio Maior

A new upper Miocene locality at Asseiceira (Rio Maior), near the top of the "Calcários de Almoster e Santarém" unit (Almoster and Santarém limestones) is studied. Animal and plant fossils are described. Comparisons are drawn to other localities related to the same unit: Freiria and Azambujeira (middle and upper levels, both with large mammals). Small mammals from Asseiceira and Freiria point out to a rather old age amidst the upper Vallesian, MN 10 mammal-unit. This gives a fairly accurate datation for the "Calcários de Almoster e Santarém" and for the short time span of the corresponding sedimentation. Climate was warm and quite dry, with contrasting seasons and arid events. During upper Vallesian times, climate in Iberian Peninsula was varied but drier than in France, and specially so in the inner basins. However in Portugal and in Catalonia climate would he less different in comparison with that of the Rhône basin. Environmental evolution has been important: at Freiria and Azambujeira (middle level) there were mainly shallow lacustrine environments that received ressurgence waters from the nearby "Maciço calcário". Humid areas were closely surrounded by dense forests and these by broader and drier savanna or steppe areas. Still later, carbonate sedimentation ended. For some time there was still a river system with oxbows; humid areas probably were decreasing in favour of surrounding, rather dry environments. This study stressed the nced for revision of the geology of the region of Rio Maior and for a new geological mapping of that area. Environmental evolution has been important: at Freiria and Azambujeira (middle level) there were mainly shallow lacustrine environments that received ressurgence waters from the nearby "Maciço calcário".

A sedimentação e a gliptogénese no registo cenozóico continental do Minho (NW Portugal)

This study identifies and describes the major Cenozoíc glyptogenesis and sedimentation episodes in the Minho region (NW Portugal). The fluvial processes of exorheic network were mainly the builders of Cenozoic landscape. This paper presents a chronological sequence of the major fluvial events based on geomorphology and sedimentary characteristics obtained in former studies and new ones. The oldest Cenozoic sedimentation (Placencian) remains on local depressions and was generated by fluvial and fluvio-lacustrine processes. Quaternary glyptogenesis begins probably with a major European cooling (Gunz?). So, the Placencian infilling was eroded and a new episode of sedimentation was accumulated in the same early paleovalley. There were three more cycles of quaternary glyptogenesis and sedimentation. The last glyptogenesis episode records the last glacial period. The post-glacial alluvion contains clasts of granites, schists, feldspats, and other alterable lithologies and minerals; on clay dimensions there are abundant illite and interstratified minerals. This composition differs from the older ones, more siliciclastic and kaolinitic. Other minor erosion episodes were identified, controlled manly by eustasy; evidences of tectonic movements were observed only in rio Minho valley.

The Neogene of Algarve (Portugal)

A synthesis about the Neogene and Quaternary of Algarve (Southern Portugal) is presented. New isotopic 87Sr/86Sr ages as well as biostratigraphic data about the Miocene deposits allow to present a new stratigraphic frame for the previously characterized units. The Lagos-Portimão Formation corresponds to deposits of temperate carbonate platform sedimentological type, developed during a long time span (Lower Burdigalian to Upper Serravallian). A major change in sedimentation conditions (carbonate to siliciclastic environments) occurred in the Lower Tortonian with the deposition of yellowish sands. Spongoliths rich in microfossils are represented in Algarve inland. Their age is not well established; calcareous nannofossils indicate the CN5a zone (Upper Serravallian) while foraminifera point out at least Nl6 zone (Lower Tortonian). In the Upper Tortonian, the sedimentation is widespread in Eastern Algarve, related with the Guadalquivir Basin infill. The deposits begin with detrital limestones, locally very rich in Heterostegina, passing to fossiliferous conglomerates and siltstones (Cacela Formation). Coarse-grained conglomerates at Galvana (Faro) pose some age problems. K/Ar age on glauconite indicates 6.72±0.17 Ma. However, glauconites may be reworked from older deposits (Cacela Formation). The Galvana Conglomerate could be related to Pliocene deposits are not well characterized. Olhos de Água sands, with a thin marine intercalation rich in marine vertebrates (fishes, a crocodile, cetaceans, sirenians), may be Upper Pliocene; however, the vertebrates point out to a Serravallian to Tortonian age. 87Sr/86Sr ages on oysters from above the level with vertebrates point out to 3.0(+2.5-1.0) Ma. Similar sand deposits occur at Morgadinho (Luz de Tavira). These sands are overlain by marls, lignite clays, lacustrine limestones and a silty calcareous crust. A small mammals association indicate an age span between Upper Pliocene and Lower Middle Pleistocene (MN17-MN20). A Biharian mammal fauna (Lower Pleistocene) was collected at Algoz in similar deposits. In the present state of knowledge, Morgadinho and Algoz deposits may be correlative.

Sedimentation in parasitological coproscopy

A sedimentation technique is described, in which a fecal suspension is placed on top of an aqueous sucrose solution of specific gravity 1.015 g/cm³. Using 100 by 15 mm test tubes, duplicate gravity sedimentation experiments were made using homogenized fecal suspensions (single-columns) and fecal suspensions placed on top of clear columns (double-columns). Egg- and cyst-counts, and turbidity determinations were made in the sediments obtained after definite time intervals. Most Ascaris lumbricoides, Trichuris trichiura and Ancylostomidae eggs sedimented within 20 minutes in single - and between 30 and 60 minutes in double-columns. Giardia duodenalis cysts required longer periods to sediment in double - than in single-columns; after 180 minutes (the maximum period of observation), double-column sediments produced 60.0% of the counts of single-columns. Double-column sediments were consistently less turbid than single-column ones

Parasitological stool sample exam by spontaneous sedimentation method using conical tubes: effectiveness, practice, and biosafety

INTRODUCTION: Spontaneous sedimentation is an important procedure for stool examination. A modification of this technique using conical tubes was performed and evaluated. METHODS: Fifty fecal samples were processed in sedimentation glass and in polypropylene conical tubes. Another 50 samples were used for quantitative evaluation of protozoan cysts. RESULTS: Although no significant differences occurred in the frequency of protozoa and helminths detected, significant differences in protozoan cyst counts did occur. CONCLUSIONS: The use of tube predicts a shorter path in the sedimentation of the sample, increases concentration of parasites for microscopy analysis, minimizes the risks of contamination, reduces the odor, and optimizes the workspace.

Oligocene sedimentation, stratigraphy, paleoecology and paleoclimatology in the big Badlands of South Dakota [by] John Clark, James R. Beerbower [and] Kenneth K. Kietzke.

v.5(1967)