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[Culture medium and grading culture technics for bioflocculant production by Paenibacillus polymyxa GA1].

A bacterial strain named GA1 which can produce bioflocculant with high flocculating activity was isolated from soil. The strain was identified as Paenibacillus polymyxa according to its morphological, physiological and biochemical characters, as well as 16S rDNA sequence (GenBank Accession number: DQ166375) similarity comparison. The results indicated that sucrose and yeast extract were the optimal carbon and nitrogen sources for bioflocculant production. Furthermore, the mass ratio of sucrose to yeast extract and the optimal sucrose concentration were ascertained. The optimum component proportion of medium (g/L) is sucrose 40.0, yeast extract 4.0, K2HPO4 5.0, KH2PO4 2.0, NaCl 0.1, MgSO4 0.2. The culture conditions including initial pH, temperature, agitation rate and inoculation quantity of strain GA1 were ascertained. Based on the relation of bacterium growth and bioflocculant production, grading culture was applied to bioflocculant production of GA1. The experimental result show that grading culture can keep high bioflocculant yield as well as shorten time of flocculant production.

Bacillus↗

[Immobilization of microorganisms on latex for producing artificial flocs].

Microorganisms belonging to the genera Pseudomonas, Micrococcus, Bacillus and Rhodotorula are flocculated with divinyl styrene latex. The process of flocculation depends on a number of physico-chemical factors but, all in all, is universal for microorganisms with different taxonomy, morphology and physiology. The structure of artificial bioflocs has been studied using optical and scanning electron microscopy.

Bacteria↗

[Comparative study on methods to recover indigenous viruses from samples of activated sewage sludge].

Samples of activated sewage sludge were investigated for their contents of indigenous enteric viruses. Therefore sewage sludge solids were collected by centrifugation and the embedded viruses eluted by blending (Waring blendor), glycine buffer (0.05 M) treatment at pH 11.5 or Freon1 treatment. Eluted viruses were concentrated on microfiberglass filters ("Balston" filters) at pH 3.5 or by flocculation with beef extract (3%, pH 3.5). Simple blending of the activated sludge was found being not very efficient, while the other two methods gave better results in freeing the viruses from the sewage flocs. Usually highest virus titers were obtained after glycine treatment and concentration with Balston filters. Freon treatment of activated sewage sludge produces aqueous supernatants containing much denaturated floc material which seems to cause a certain number of viruses to remain in the supernatant after flocculation.

Centrifugation↗

Mapping of the FLO5 gene in saccharomyces cerevisiae.

The FLO5 gene has been assigned to chromosomes I and VIII in Saccharomyces cerevisiae. According to the results obtained, flocculation of strain STX347-ID (reported as strain FLO5) is the result of two different genes: one of them is allelic to the previously reported FLO1 and located at 4.7 cM from PHO11 and 37 cM from ADE1 on chromosome I. The other was mapped on chromosome VIII at 36.8 cM from PET3 and 30.5 cM from FUR1. This second gene generated a constitutive flocculation phenotype that was not under the control of the fsu3 suppressor. The latter has been reported to suppress certain FLO1 genes.

Alleles↗

Nonflocculent versus total biomass ratio as a criterion for starting the biomass separation process

We introduce the ratio of nonflocculent versus total biomass as a criterion for starting cell separation from the medium. This criterion can be applied for the automation of the process regardless of the process dynamics. Its minimum indicates the optimum period of time for the start of the separation process with regard not only to nonflocculent cell concentration, but also medium attributes. In contrast to the concentration of nonflocculent cells, which has two minima, first at the beginning of the process and another broader one in the period during which maximum flocculation is present, the ratio has a single minimum and can therefore be implemented as a criterion for cell separation. To calculate the ratio value, in addition to an on-line method for nonflocculent biomass measurement described elsewhere, an on-line method for the total biomass of flocculent yeast is proposed. It is based on the absorbency measurement of the cell biomass, previously deflocculated by EDTA. Therefore, it can be applied in bioprocesses with transparent media and yeast that can be deflocculated by EDTA. Copyright 1998 John Wiley & Sons, Inc.

Journal Article↗

Postsynaptic actin filaments at the giant mossy fiber-unipolar brush cell synapse.

The unipolar brush cell (UBC), a small interneuron occurring at high density in the granular layer of the mammalian vestibulocerebellum, receives a giant glutamatergic synapse from a single mossy fiber (MF) rosette, usually on a brush of dendritic branchlets. MF stimulation produces a current in the UBC several orders of magnitude greater in duration than at other glutamatergic synapses. We assumed that the cytoskeleton would have a special role in plasticity of the MF-UBC synapse. Neurofilaments and microtubules are enriched in the UBC somatodendritic compartment but are conspicuously absent in close proximity to the giant synapse, where standard electron microscopy reveals a granulo-flocculent material. Because osmium tetroxide fixation during sample preparation for standard electron microscopy destabilizes actin filaments, we hypothesized that this subsynaptic granulo-flocculent material is actin-based. After actin stabilization, we observed prominent, but loosely organized, bundles of microfilaments at the subsynaptic region of the MF-UBC synapse that linked the postsynaptic density with the cytoskeletal core of the dendritic branchlets. Confocal fluorescence microscopy and pre- and postembedding immunogold labeling with phalloidin and actin antibodies showed that these microfilaments consist of f-actin and contain little beta-actin. This extraordinary postsynaptic actin apparatus is ideally situated to form a dynamic framework for glutamate receptors and other postsynaptic molecules, and to mediate activity-dependent plastic rearrangements of the giant synapse.

Actin Cytoskeleton↗

Role of epithelial clear cells of the rat epididymis in the disposal of the contents of cytoplasmic droplets detached from spermatozoa.

Upon release from the seminiferous epithelium, spermatoza show a small droplet of cytoplasm attached to the neck region. During transit of spermatozoa in the caput epididymidis, this cytoplasmic droplet migrates along the middle piece of the flagellum. In the corpus epididymidis, the droplet shows a lateral displacement, while in the cauda epididymidis it detaches from the spermatozoon. In the electron microscope, cytoplasmic droplets attached to spermatozoa were seen to contain numerous, short, straight or C-shaped, flattened membranous elements referred to as lamellae, small vesicles, and small particles (35-nm diameter) with a diffuse wall showing no apparent unit membrane. The lamellae were stacked closely on one another or arranged in a loose array. Structurally as well as cytochemically, with different cytochemical markers, the lamellae and vesicular elements failed to show any evidence of being components of the Golgi apparatus or elements of the endoplasmic reticulum. The lamellae, vesicular elements, and 35-nm particles were also seen free in the lumen of the corpus epididymidis but were especially prominent in the cauda epididymidis at a time when droplets were being released from spermatozoa. The lumen of the epididymis, as spermatozoa passed from the caput to the cauda epididymidis, was also noted to acquire progressively a flocculent background material. The epididymal epithelium is composed predominantly of principal and clear cells. The endocytic activity of clear cells was examined in rats at different time intervals after a single injection of cationic ferritin into the lumen of the cauda epididymidis. At 2 min the tracer was bound to the microvilli of these cells and was also observed within large coated and uncoated pits, subsurface coated vesicles, and numerous subsurface small uncoated vesicular membranous elements (150-200-nm diameter). At 5 min, in addition to the above structures, the tracer was present in endosomes, while at 15 and 30 min, pale and dense multivesicular bodies appeared labeled, respectively. At 1 and 2 hr, but more so at 6 hr large dense membrane-bound bodies identified cytochemically as secondary lysosomes became labeled. All of the above endocytic structures were also seen to contain the 35-nm particles, flattened or vesicular membranous profiles, and a fine flocculent background material reminiscent of those seen free in the lumen or found in cytoplasmic droplets attached to spermatozoa. (ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Improved ultrastructural preservation of epiphyseal chondrocytes by the freeze-substitution method.

The ultrastructure of epiphyseal chondrocytes was studied following quick-freezing and freeze-substitution, and was compared to that of cells fixed with aqueous aldehydes. The former approach provided an improved ultrastructural preservation whereby every type of chondrocyte exhibited a smoother cell contour. The plasma membrane as well as intracytoplasmic membranes revealed a trilaminar substructure. The intracytoplasmic ground substance was composed of flocculent materials which were in direct contact with the inner leaflet of the plasma membrane. Within the extracellular matrix the proteoglycan network adhered to the outer leaflet of the plasma membrane. Whenever cellular shrinkage took place, the flocculent matrix within the cytoplasm and the proteoglycan network in the pericellular matrix disappeared. The contents of the RER, the Golgi apparatus, and the intracellular vesicles and vacuoles were well retained. In the proliferative zone, the Golgi saccules of young cells contained a thread-like structure showing a clear periodicity. The cytoplasmic vesicles and vacuoles showed marked variation in their electron density. Intramitochondrial granules were sensitive to aqueous treatments, as evidenced by the observation that they disappeared after either floating on water or staining with aqueous solution. In the calcifying zone, mitochondrial granules were noted within hypertrophic chondrocytes, a feature that was not observed following conventional processing. Cytoskeletal elements were well preserved in all types of cells. A dense microfilamentous network occupied the pericellular cytoplasm. Bundles of microfilaments were seen in the cellular peripheral processes. Microtubules were distributed throughout the cytoplasm, and the Golgi complex was intimately associated with the microtubule network; it appears that the secretory processes are involved with the microtubules.

Animals↗

Synaptic contacts on glial cells in the abdominal ganglion of Aplysia californica.

Two types of glial cells have been identified in the neuropil and in the subcapsular space of the abdominal ganglion of Aplysia californica. In the neuropil, a first type of glial cell has a light staining cytoplasm and gives rise to processes which may contain bundles of filaments, a flocculent material or bundles of microtubules and membranous sacs. Synaptic contacts have been seen very rarely on their somata and more frequently on their "filamentous" and "flocculent" processes. A second type of glial cell has a darkly staining cytoplasm and gives rise to dense processes. Synaptic contacts are frequently seen on these processes. In the subcapsular zone, there are also two types of glial cells, one with a clear and the other with a dense cytoplasm. The processes of the clear glial cells contain glycogen-like particles and sometimes bundles of filaments. These processes surround the bundles of neuronal profiles which form the perisomatic plexus. Such enveloping processes receive synaptic contacts from the neuronal profiles in the bundles. The dense glial cells of the subcapsular zone also receive synaptic contacts, but this is a rare occurrence.

Animals↗

Aspiration cytology diagnosis of a primary amyloid tumor of thoracic vertebra: a case report.

Isolated primary amyloid tumor of bone is rare; however, preoperative diagnosis can be rewarding because the prognosis is excellent. There are no clinical or roentgenographic criteria that can establish this diagnosis. There are few previous reports of fine-needle aspiration (FNA) where diagnosis of amyloidoma was made retrospectively. They describe the presence of extracellular hyaline material along with plasma cells and lymphocytes. A 35-year-old female was referred to the FNA clinic with swelling in the right upper chest wall. Radiologic findings revealed a destructive lytic lesion involving the bodies of D1 and D2 vertebrae with extension into surrounding soft tissue. Repeated FNA smears were hypocellular but had abundant homogeneous flocculent material, which stained positive with Congo red. A few plasma cells and foreign-body giant cells were also seen. We conclude that preoperative FNA cytology diagnosis of amyloid tumor is possible. Hypocellular smears with flocculent material, plasma cells, and foreign-body giant cells in absence of granulation tissue should suggest the diagnosis.

Adult↗

Elaboration of an oviductin by the oviductal epithelium in relation to embryo development as visualized by immunocytochemistry.

The hamster oviduct secretes a high molecular weight antigen that belongs to the family of glycoproteins known as oviductins. In the present study, using immuno-electron microscopy, we examined the location of this hamster oviductin-1 (Hm Ov-1) in hamster oviductal oocytes and early embryos up to the blastocyst stage. The immunoreactive pattern of Hm Ov-1 changes markedly during the embryo development. In oviductal oocytes prior to fertilization, Hm Ov-1 was associated exclusively with the zona pellucida. Following fertilization, immunolabeling was detected in the perivitelline space and over the plasma membrane of 2-cell, 4-cell, and 8-cell embryos as well as young blastocysts. The change of the immunoreactive pattern was accompanied by the formation of an abundant number of coated pits, endocytic vesicles, multivesicular bodies, and lysosomal-like structures which were strongly labeled by gold particles. These immunogold-labeled cytoplasmic organelles characteristic of the endosomal-lysosomal apparatus were particularly evident in 2-cell, 4-cell, and 8-cell embryos and showed a decrease in number in the blastocysts. The close resemblance between the labeled flocculent material detected in the perivitelline space and that found in the zona matrix of early embryos and blastocysts suggested that the Hm Ov-1-associated electron-dense, flocculent material in the perivitelline space originated from the zona pellucida and was later endocytosed by the blastomeres through coated pits and endocytic vesicles. The detection of Hm Ov-1 in numerous multivesicular bodies and lysosomal structures indicated that the oviductin is eventually degraded. Although the exact functional role of Hm Ov-1 is not known, the presence of a copious amount of Hm Ov-1 in early hamster embryos may be ascribed to a special relationship between this particular oviductin and embryo development.

Animals↗

Interaction of xanthan gum with suspended solids.

Xanthan gum was adsorbed significantly by magnesium carbonate, aluminum hydroxide, zinc oxide, and calcium carbonate, giving Langmuir-type isotherms. Saturation adsorption was higher from 0.9% NaCl than from water due to reduced mutual repulsion of polymer segments in the presence of the salt. Adsorption resulted from electrostatic attraction between positively charged particles and the negatively charged polymer. zeta-Potential measurements correlated with the adsorption data but were not predictive of the flocculation state. The result indicate that flocculation of magnesium carbonate and aluminum hydroxide by xanthan gum is consistent with a bridging mechanism.

Adsorption↗

Conductivity studies of suspension systems in different states of aggregation.

The electrical conductivity effects of dispersed, coagulated, and flocculated systems were investigated using sulfamerazine powder, an insoluble, hydrophobic drug to prepare the suspension systems. For the dispersed systems, a peak in conductivity was observed at a drug concentration between 5 and 15%. The critical coagulating concentration was defined as the concentration of drug at which a maximum in specific conductance was observed. At this concentration, a maximum number of charged particles were in the system. Coagulated suspensions showed higher conductance values than the dispersed systems at equivalent concentrations; however, the critical coagulating concentration value appeared to be the same. For flocculated suspensions there was an increase in conductance with drug concentration with no perceptible peak conductance value.

Chemistry, Pharmaceutical↗

Dissolution dialysis studies of metronidazole-montmorillonite adsorbates.

In the context of the potential usefulness of clays in retarding the rate of release of adsorbed drugs, dissolution dialysis studies of the release of metronidazole from montmorillonite adsorbates have been conducted. The goal was to develop a means for improving local gastrointestinal therapy of amebiasis while concurrently maintaining efficacy in treating hepatic amebiasis. At acidic pH, the clay was in a flocculated state and the rate of drug release was inhibited. This effect was apparently due to slow diffusion of the drug throughout the clay flocculate. A physical admixture of montmorillonite and metronidazole was also effective in inhibiting the rate of release of metronidazole. Upon increasing the pH to 7, the clay particles progressively deflocculated and the rate of release increased significantly.

Adsorption↗

Combined enzyme histochemical and ultrastructural study on cryostat sections of pig heart to detect early reperfusion damage after ischaemia.

In cardiac surgery, recognition of peroperative myocardial infarction may improve patient selection for prolonged circulatory support. The value of enzyme histochemistry to discriminate between reversible and irreversible myocardial damage at short periods of reperfusion was studied in an in vivo model of regional ischaemia in pig hearts. The left anterior descending coronary artery (LADCA) was clamped for 45 min followed by 2 h reperfusion (group 1, n = 3). Post-mortem heart tissue showed markedly decreased activities of lactate dehydrogenase (LDH) and beta-hydroxybutyrate dehydrogenase (BDH) as demonstrated in cryostat sections, accompanied by massive leakage of LDH in the venous effluent. The depleted areas showed irreversible cell damage based on the presence of flocculent densities in mitochondria. In group 2 (n = 6), LADCA flow was reduced to 40 per cent of the base-line value followed by 2 h reperfusion. Heart tissue showed normal LDH and BDH activities, except for some cells surrounding blood vessels, which activity was minimally decreased. Flocculent densities in mitochondria were never observed. We conclude that enzyme histochemistry of LDH and BDH activity on cryostat sections is a useful tool for detecting irreversible myocardial cell damage as early as 2 h reperfusion after ischaemia of the pig heart. The technique has potential applications in the detection of peroperative infarction in human biopsies.

Animals↗

Biological treatment of wastewater from the cassava meal industry.

Biological treatments of cassava meal processing wastewaters were investigated by aerobic and combined anaerobic/aerobic reactors. As a pretreatment, flocculation and sedimentation with aluminum salts and natural polyelectrolytes were employed, in order to change the effluent concentration of organics from 14,000 to 2000 mg/L in the bench scale reactor, with a hydraulic retention time of 37 min and an influent rate of 0.56 L/cm. Biological degradation by an activated sludge process was carried out using a reactor volume of 20 L. The observed organic matter removal rates were 89 to 93%, the cyanide removal was 95 to 99%, and the food to microorganisms ratio was found to be in the range of 0.166 to 0.242/day, with a hydraulic residence time of 1.4 to 4.2 days. As a posttreatment, flocculation and coagulation were employed, resulting in an effluent of good quality, as shown by turbidity measurements and negative tests for fecal and total coliforms.

Bacteria, Anaerobic↗

Micromechanics: A New Approach to Studying the Strength and Breakup of Flocs

Micromechanical techniques were developed to pull apart single floc particles 6 to 40 μm in size in order to measure floc strengths and fractal dimensions. Flocs were formed by aggregating aqueous precipitated calcium carbonate sols with two water-soluble polymers. The tensile strengths ranged from 20 to 200 nN and showed no correlation with floc sizes when plotted on logarithmic scales. Flocs created by the two flocculant systems were found to have fractal dimensions of 1.8 and 2.4 based on analysis of fragment sizes. The compact flocs favored breakup by surface erosion, whereas flocs characterized by the lower fractal dimension tended to undergo large scale fragmentation. Comparison of the two polymeric flocculants revealed that both the fractal dimension and the tendency for ruptured fragments to reflocculate were sensitive to the polymer used.

Journal Article↗

Stability of Concentrated Colloids: The Controlling Parameters.

A theoretical framework has been developed for predicting and explaining the colloidal stability of concentrated dispersions. It is based on multiparticle interaction models for the electrical double layer (EDL) and van der Waals (vdW) interactions. An asymmetric cell model is used for estimating EDL interaction energy from the solution of the Poisson-Boltzmann equation. The unretarded vdW interaction energy is calculated via Hamaker's method, adopting the pairwise-additivity approach. Various dispersion parameters are investigated for their effects on the flocculation energy barrier. These include volume fraction, size, and surface potential of particles, as well as the ionic strength of the dispersing medium. The results show several new trends borne out of multiparticle interaction effects. A maximum in the energy barrier is predicted as a function of salinity, contrasting the monotonic dependence from the classical DLVO theory. This salinity dependence of the energy barrier provides the first comprehensive explanation of reported experimental findings on stability and rheological properties of concentrated dispersions. Other predictions include the occurrence of flocculation at a secondary minimum and the particle-size dependent variation of suspension stability with particle volume fraction. Copyright 1998 Academic Press.

Journal Article↗