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PubMed2003

Determination of some rheological parameters for the characterization of activated sludge.

The rheological characterization of activated sludge originating from different aeration tanks and from a lab-scale plant was carried out using rotation tests. Two rheological parameters were used: limit viscosity (mu(infinity)) and reduced hysteresis area (rHa). Mu(infinity) was strongly influenced by the total suspended solids (TSS) content of sludge and an exponential relation was found between TSS and the rheological parameter mu(infinity). Significant differences in sludge viscosity were found for sludge of different origins, but all of 10 g/l TSS content, indicating ability of viscosity to show different sludge qualities. Marked changes in activated sludge microorganisms communities were shown to have an influence on rheological parameters. During aerobic digestion of sludge, variations in mu(infinity) were noticed, indicating change of sludge quality. An over-growth of filamentous bacteria species was shown to induce a particular rheological behavior which could be characterized by rHa. This parameter was proposed as an index of filamentous bacteria proliferation in activated sludge aeration tanks.

Bacterial Physiological Phenomena↗
PubMed2006

Meganema perideroedes gen. nov., sp. nov., a filamentous alphaproteobacterium from activated sludge.

An industrial wastewater treatment plant at Grindsted, Denmark, has suffered from bulking problems for several years caused by filamentous bacteria. Five strains were isolated from the sludge by micromanipulation. Phylogenetic analysis of the 16S rRNA gene sequences showed that the strains formed a monophyletic cluster in the Alphaproteobacteria, and they were phenotypically different from their closest relatives and from all hitherto known filamentous bacteria described (closest relative Brevundimonas vesicularis ATCC 11426(T), 89.8 % sequence similarity). In pure culture, the cells (1.5-2.0 microm) in filaments are Gram-negative and contain polyphosphate and polyhydroxyalkanoates. The optimum temperature for growth is 30 degrees C and the strains grow in 2 % NaCl and are oxidase- and catalase-positive. Ubiquinone 10 is the major quinone. The major fatty acid (C(18 : 1)omega7c) and smaller amounts of unsaturated fatty acids, 3-hydroxy fatty acids with a chain length of 16 and 18 carbon atoms and small amounts of 10-methyl-branched fatty acids with 18 carbon atoms (C(19 : 0) 10-methyl) affiliated the strains with the Methylobacterium/Xanthobacter group in the Alphaproteobacteria. The G+C content of the DNA is 42.9 mol% (for strain Gr1(T)). The two most dissimilar isolates by 16S rRNA gene comparison (Gr1(T) and Gr10; 97.7 % identical) showed 71.5 % DNA-DNA relatedness. Oligonucleotide probes specific for the pure cultures were designed for fluorescence in situ hybridization and demonstrated that two filamentous morphotypes were present in the Grindsted wastewater treatment plant. It is proposed that the isolates represent a new genus and species, Meganema perideroedes gen. nov., sp. nov. The type strain of Meganema perideroedes is strain Gr1(T) (=DSM 15528(T)=ATCC BAA-740(T)).

Alphaproteobacteria↗
PubMed1997

Cell-specific respiratory activity of aquatic bacteria studied with the tetrazolium reduction method, cyto-clear slides, and image analysis.

We present an improvement of the INT [2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride)] reduction method using Cyto-Clear slides, the fluorochrome DAPI (4(prm1),6(prm1)-diamidino-2 phenylindole), and an image analysis system. With this method we were able to simultaneously measure cell dimensions and formazan crystals as indicators of the respiratory activity of single bacteria. The method was tested on a natural bacterioplankton community of an oligotrophic high mountain lake (Gossenkollesee, Tyrolean Alps, Austria, 2,417 m above sea level) in midwinter ((symbl)1-m-thick ice and snow layer; dissolved organic carbon, 0.51 mg liter(sup-1); water temperature, 2(deg)C). About 25% of planktonic bacteria were respiratorily active, and a complex pattern of bacterial morphologies and specific respiratory activities was observed during a time series of INT incubation. Rod-shaped bacteria with cell lengths of between 1.6 and 4.8 (mu)m already showed visible activity after 0.5 h of INT incubation. Small cells (rods and cocci) in the size fraction <1.6 (mu)m and long filamentous bacteria (up to 120 (mu)m) were visibly active only after a 2-h incubation period. After 8 h of incubation, more than 90% of all cells between 3.2 and 6.4 (mu)m in cell length were respiratorily active, whereas only 5% of cells <1.6 (mu)m and 50% of filamentous bacteria contained formazan grains. We could distinguish five major bacterial phenotypes that showed distinct activity patterns with respect to incubation period and numbers and sizes of formazan crystals. There was no correlation between the total formazan volume per active cell and bacterial cell volume, and for any size class of active bacteria, total formazan volumes varied by about 2 orders of magnitude after 8 h of incubation. This indicates that cell-specific activity is extremely variable and is not related to size and that a small portion of all cells may account for the overall activity.

Journal Article↗
PubMed1992

Effect of temperature and medium composition on mycelial growth of Streptomyces tendae in submerged culture.

To investigate the influence of culture conditions on growth of filamentous bacteria in submerged culture, S. tendae was cultivated at various temperatures and in various media. For this purpose, a temperature-controlled growth chamber was constructed, which allows the cultivation of filamentous bacteria and fungi oxygen saturated medium. To observe the development of mycelia emerging from spores, this growth chamber was mounted on a microscope stage and series of images were analyzed by an image processing system. Growth kinetics obtained in liquid culture were identical to those determined on solid media. Specific growth rate and apical extension rates of individual hyphae seemed to be higher than those observed on solid media, due to a better supply of medium in submerged culture. A two-fold increase in specific growth rate, mean apical extension rate and branching rate was observed when the temperature was increased from 27 degrees C to 37 degrees C, therefore, the length of the hyphal growth unit was not changed. Growth in synthetic media used here was considerably slower compared with complex medium. A decrease in glucose concentration from 40.0 g l-1 to 1.0 g l-1 resulted in an increase in branching rate and specific growth rate, while apical extension rate of individual hyphae was unchanged.

Bacteriological Techniques↗
PubMed2006

Pathological findings in the lens capsules and intraocular lens in chronic pseudophakic endophthalmitis: an electron microscopy study.

PURPOSE: The aim of this study was to describe the pathological findings in lens capsules and intraocular lens (IOL) studied by scanning and/or transmission electron microscopy (SEM and TEM, respectively) in a series of four eyes with chronic pseudophakic endophthalmitis (CPE). PATIENTS AND METHODS: We performed a retrospective study of four patients presenting CPE in whom surgical treatment with pars plana vitrectomy, capsulectomy with extraction of the IOL, and intravitreous antibiotic therapy was thereafter performed. The extracted IOL and the capsular remains were studied by SEM and/or TEM and microbiologic analysis of aqueous humour and vitreous aspirate was also carried out in all the cases. RESULTS: The presence of microorganisms was observed in the material analysed in all the cases studied. The use of TEM identified bacterial contamination by Staphylococcus spp and mixed contamination with microorganisms presenting a bacillar morphology suggestive of infection by Propionibacterium acnes in addition to the presence of cocci in the capsular remains. In another two cases, SEM localized colonies of Staphylococcus spp on the surface of the IOL in one case and mixed bacterial colonization with cocci plus filamentous bacteria in the other. The presence of macrophages associated with bacteria was observed in the capsular remains. CONCLUSIONS: Microorganisms were found in the IOL or the capsular material in the four cases studied, thereby explaining the refractoriness and severity of infection. The possible presence of polymicrobial infections, especially in the cases with filamentous bacteria, also explains the recurrence of infection.

Aged↗
PubMed1998

Acid fast filaments in stool samples from an AIDS patient.

The presence of filamentous bacteria morphologically similar to Nocardia in a fresh stool sample from an AIDS patient with pulmonary nocardiosis is here reported. The material was submitted to our laboratory for a parasitologic examination and was stained by the Kinyoun method, revealing numerous delicate, irregularly stained, branching acid-fast filaments. Nocardia asteroides had been isolated from sputum samples of this patient. The patient was a 32 year-old HIV+ female admitted to our center on June 1997 because of productive cough, right-sided thoracic pain and weight loss. Chest X rays showed the presence of right superior lobe excavated pneumonia. This was the first time we had observed filamentous bacteria similar to Nocardia in a stool sample submitted to parasitologic examination. For similar cases, and when its presence was not detected in other specimens collected from the same patient, intestinal endoscopy and biopsy should be performed for eventual lesions and smear examination repeated with Kinyoun stain and cultures for Nocardia.

AIDS-Related Opportunistic Infections↗
PubMed2003

Performance and microbial dynamics in the coarse pore filtration activated sludge process at different SRTs (solids retention times).

In this research, three SRTs (about 10, 30 and 75 days (without wasting the sludge except for sampling)) were applied to three reactors equipped with non-woven and coarse pore filter modules. The flux was adjusted to about 1 m/d during operation. The main objective of the study was to compare the performance and microbial population dynamics under different SRTs in this process. The results of reactors with SRTs of about 10 and 30 days have shown very good effluent quality without any clogging problem for more than 4 months of operation. For the reactor with long SRT (75 days), the filter clogging was observed after about 80 days of operation and caused an increase in the operation pressure and deterioration effluent quality on some days. Excessive abundance of filamentous bacteria was observed in the reactor with SRT of about 10 days, which had the best effluent quality. According to the FISH results, type 021N was predominant in the reactor with long SRT, which had the clogging problem. On the other hand, other reactors (with SRTs of about 10 and 30 days) did not contain much type 021N, but some other filamentous bacteria dominated. Maximum EPS concentration (as mg/L) was observed in the reactor with long SRT. Also the abundance of two types of metazoa (Pristina sp. and tardigrades) was observed in the reactor with long SRT, which had the clogging problem and poor effluent quality.

Animals↗
PubMed2005

Isotope labeling and microautoradiography of active heterotrophic bacteria on the basis of assimilation of 14CO(2).

Most heterotrophic bacteria assimilate CO(2) in various carboxylation reactions during biosynthesis. In this study, assimilation of (14)CO(2) by heterotrophic bacteria was used for isotope labeling of active microorganisms in pure cultures and environmental samples. Labeled cells were visualized by microautoradiography (MAR) combined with fluorescence in situ hybridization (FISH) to obtain simultaneous information about activity and identity. Cultures of Escherichia coli and Pseudomonas putida assimilated sufficient (14)CO(2) during growth on various organic substrates to obtain positive MAR signals. The MAR signals were comparable with the traditional MAR approach based on uptake of (14)C-labeled organic substrates. Experiments with E. coli showed that (14)CO(2) was assimilated during both fermentation and aerobic and anaerobic respiration. The new MAR approach, HetCO(2)-MAR, was evaluated by targeting metabolic active filamentous bacteria, including "Candidatus Microthrix parvicella" in activated sludge. "Ca. Microthrix parvicella" was able to take up oleic acid under anaerobic conditions, as shown by the traditional MAR approach with [(14)C]oleic acid. However, the new HetCO(2)-MAR approach indicated that "Ca. Microthrix parvicella," did not significantly grow on oleic acid under anaerobic conditions with or without addition of NO(2)(-), whereas the addition of O(2) or NO(3)(-) initiated growth, as indicated by detectable (14)CO(2) assimilation. This is a metabolic feature that has not been described previously for filamentous bacteria. Such information could not have been derived by using the traditional MAR procedure, whereas the new HetCO(2)-MAR approach differentiates better between substrate uptake and substrate metabolism that result in growth. The HetCO(2)-MAR results were supported by stable isotope analysis of (13)C-labeled phospholipid fatty acids from activated sludge incubated under aerobic and anaerobic conditions in the presence of (13)CO(2). In conclusion, the novel HetCO(2)-MAR approach expands the possibility for studies of the ecophysiology of uncultivated microorganisms.

Actinobacteria↗
PubMed1996

Characterization of Bacterial Communities from Activated Sludge: Culture-Dependent Numerical Identification Versus In Situ Identification Using Group- and Genus-Specific rRNA-Targeted Oligonucleotide Probes

The structures of bacterial communities were studied in activated sludge samples obtained from the aerobic and anaerobic zones of a wastewater treatment plant showing enhanced phosphorous removal. Samples were analyzed by in situ hybridization with oligonucleotide probes complementary to selected regions of the 16S and 23S ribosomal RNA (rRNA) characteristic for defined phylogenetic entities (genera and larger groups). The microbial community structures revealed by molecular techniques were compared with the compositions of culturable bacterial communities, obtained from the characterization of 255 isolates from tryptone-soy (TS) agar and R2A agar. These isolates were characterized by 89 physiological tests and their cellular fatty acid patterns, and identified. Culture-dependent techniques indicated the following distribution: different Aeromonas spp. (2.7-8.3% on R2A agar; 45.0-63.7% on TS agar), Acinetobacter spp. (5.4-9.0% on R2A agar; 5.0-9.1% on TS agar), Pseudomonas spp. (up to 10% on R2A agar) and Shewanella putrefaciens (up to 3.0% on R2A agar), all members of the gamma subclass of Proteobacteria, were isolated most frequently. The relatively rare isolates of the beta subclass were identified as Acidovorax spp., Alcaligenes spp., and Comamonas spp.. The Gram-positive bacteria (high DNA G+C) were assigned mainly to Arthrobacter spp., Microbacterium spp., and Mycobacterium phlei. In order to assess the in situ abundance of the most frequently isolated genus, Aeromonas, two rRNA-targeted oligonucleotide probes were developed. The two gamma proteobacterial genera Aeromonas and Acinetobacter constituted less than 5% of all bacteria. In situ, Proteobacteria belonging to the beta subclass and high G+C Gram-positive bacteria were dominant. From filamentous bacteria, Sphaerotilus spp. and Leptothrix spp. could be detected occasionally. In addition, one sample contained a high proportion of the morphologically distinct filaments of Microthrix parvicella.

Journal Article↗
PubMed2000

Evaluation of the SOS/umu-test post-treatment assay for the detection of genotoxic activities of pure compounds and complex environmental mixtures.

This study presents an evaluation of the SOS/umu-test after introducing an additional dilution and incubation in the post-treatment assay. This treatment reduces the influence of coloured test compounds that otherwise affect the colorimetric determination of the beta-galactosidase activity and the bacterial growth measurement during the testing of complex environmental samples. The post-treatment assay significantly increased the beta-galactosidase activity and consequently the enzyme induction ratios at higher doses of model genotoxins 4-nitroquinoline-N-oxide, N-methyl-N'-nitro-N-nitrosoguanidine, 2-aminoanthracene, benzo(a)pyrene with low or no effect on the sensitivity of the test itself. On the other hand tests of environmental extracts indicated significant increases in sensitivity after additional incubation. 4-Nitroquinoline-N-oxide treatments of bacteria in the test affected cell division and caused filamentous growth. The size of filamentous bacteria and incidence rate of the length categories was positively correlated with the concentrations of genotoxins. Presence of filamentous tester bacteria proved induction of SOS response and genotoxic activity of environment samples in SOS/umu-test.

4-Nitroquinoline-1-oxide↗
PubMed1999

Phylogenetic relationships of filamentous sulfur bacteria (Thiothrix spp. and Eikelboom type 021N bacteria) isolated from wastewater-treatment plants and description of Thiothrix eikelboomii sp. nov., Thiothrix unzii sp. nov., Thiothrix fructosivorans sp. nov. and Thiothrix defluvii sp. nov.

The relationship of mixotrophic and autotrophic Thiothrix species to morphologically similar chemoorganotrophic bacteria (e.g. Leucothrix species, Eikelboom type 021N bacteria) has been a matter of debate for some years. These bacteria have alternatively been grouped together on the basis of shared morphological features or separated on the basis of their nutrition. Many of these bacteria are difficult to maintain in axenic culture and, until recently, few isolates were available to allow comprehensive phenotypic and genotypic characterization. Several isolates of Thiothrix spp. and Eikelboom type 021N strains were characterized by comparative 16S rRNA sequence analysis. This revealed that the Thiothrix spp. and Eikelboom type 021N isolates formed a monophyletic group. Furthermore, isolates of Eikelboom type 021N bacteria isolated independently from different continents were phylogenetically closely related. The 16S rRNA sequence-based phylogeny was congruent with the morphological similarities between Thiothrix and Eikelboom type 021N. However, one isolate examined in this study (Ben47) shared many morphological features with the Thiothrix spp. and Eikelboom type 021N isolates, but was not closely related to them phylogenetically. Consequently, morphology alone cannot be used to assign bacteria to the Thiothrix/type 021N group. Comparative 16S rRNA sequence analysis supports monophyly of the Thiothrix/type 021N group, and phenotypic differences between the Thiothrix spp. and Eikelboom type 021N bacteria are currently poorly defined. For example, both groups include heterotrophic organisms that deposit intracellular elemental sulfur. It is therefore proposed that the Eikelboom type 021N bacteria should be accommodated within the genus Thiothrix as a new species, Thiothrix eikelboomii sp. nov., and three further new Thiothrix species are described: Thiothrix unzii sp. nov., Thiothrix fructosivorans sp. nov. and Thiothrix defluvii sp. nov.

Base Composition↗
PubMed1986

Ultrastructural development of dated plaque in case of denture stomatitis.

Using special acrylic discs glued into the palatal surface of a denture the development of denture plaque in a case of denture stomatitis as well as the relationships between the denture surface, pellicle, plaque and palatal epithelium were studied with transmission electron microscopy after 30 mn, 1 h, 4 h, 8 h, 24 h, 48 h, 9 days and 29 days. A thin pellicle which increased in thickness from 9 hours to 28 days was visible at the surface of the acrylic denture. It appeared that bacteria retained in the palatal epithelial intercellular spaces were the source of the plaque which developed at the surface of the acrylic denture. Initially loosely packed cocci-like and a few rod-shaped Gram positive bacteria appeared in 8 hours samples at the denture-epithelium interface. An important increase in denture plaque thickness was noted between 24 and 48 hours. In 9 and 28 days samples, coccoid and rod-shaped Gram positive and Gram negative bacteria were present and filamentous bacteria began to be apparent. In all the samples studied, Candida was rare.

Acrylic Resins↗
PubMed2001

Chlorine-susceptible and chlorine-resistant type 021N bacteria occurring in bulking activated sludges.

Two filamentous bacteria causing bulking in two activated sludges were examined. Investigations using morphological features, staining techniques, and fluorescent in situ hybridization identified both filaments as type 021N. However, an examination of the effect of chlorine on the sludges revealed a chlorine-susceptible type 021N in one sludge and a chlorine-resistant type 021N in the other.

Chlorine↗
PubMed2005

Structural and spectroscopic properties of a reaction center complex from the chlorosome-lacking filamentous anoxygenic phototrophic bacterium Roseiflexus castenholzii.

The photochemical reaction center (RC) complex of Roseiflexus castenholzii, which belongs to the filamentous anoxygenic phototrophic bacteria (green filamentous bacteria) but lacks chlorosomes, was isolated and characterized. The genes coding for the subunits of the RC and the light-harvesting proteins were also cloned and sequenced. The RC complex was composed of L, M, and cytochrome subunits. The cytochrome subunit showed a molecular mass of approximately 35 kDa, contained hemes c, and functioned as the electron donor to the photo-oxidized special pair of bacteriochlorophylls in the RC. The RC complex appeared to contain three molecules of bacteriochlorophyll and three molecules of bacteriopheophytin, as in the RC preparation from Chloroflexus aurantiacus. Phylogenetic trees based on the deduced amino acid sequences of the RC subunits suggested that R. castenholzii had diverged from C. aurantiacus very early after the divergence of filamentous anoxygenic phototrophic bacteria from purple bacteria. Although R. castenholzii is phylogenetically related to C. aurantiacus, the arrangement of its puf genes, which code for the light-harvesting proteins and the RC subunits, was different from that in C. aurantiacus and similar to that in purple bacteria. The genes are found in the order pufB, -A, -L, -M, and -C, with the pufL and pufM genes forming one continuous open reading frame. Since the photosynthetic apparatus and genes of R. castenholzii have intermediate characteristics between those of purple bacteria and C. aurantiacus, it is likely that they retain many features of the common ancestor of purple bacteria and filamentous anoxygenic phototrophic bacteria.

Amino Acid Sequence↗
PubMed2006

Microbial community change of sulfate reduction and sulfur oxidation bacteria in the activated sludge cultivated with acetate and peptone.

The growth of sulfate reducing bacteria (SRB) and filamentous sulfur bacteria was monitored on a laboratory scale in activated sludge reactors using acetate and peptone as the artificial wastewater. When the artificial wastewater contained acetate and peptone, filamentous bacteria increased in the sludge and the SVI values increased. There was a good correlation between sulfate reducing activity and sulfur oxidation activity in the produced sludge. The microbial community change of filamentous sulfur bacteria and sulfate reducing bacteria was analyzed using the fluorescent in situ hybridization (FISH) method. The tendency for the growth of filamentous sulfur bacteria Thiothrix eikelboomii following the growth of SRB was observed. The percentage of SRB385- hybridized cells and DNMA657-hybridized cells found in the total cell area increased from 2-3% to 7-10% when the filamentous bulking occurred.

Acetates↗
PubMed1978

Ultrastructure of the subgingival microflora in juvenile periodontitis.

The ultrastructure of the subgingival deposits on the root surfaces of teeth affected by juvenile periodontitis was studied on 12 teeth from nine individuals, 15--30 years of age. The deposits consisted of either microbial masses associated with a pellicle, or of a cuticular material almost free of bacteria. Gram-negative rods and filaments were the predominant microorganisms. "Corncob" configurations consisting of filamentous bacteria surrounded by Gram-positive cocci, and "bristle brush" formations comprising corncobs surrounded by long rods were observed in the superficial layer of the plaque. Spirochetes and flagellated rods constituted a major segment of the microflora. The present data indicated that the deep pockets in juvenile periodontitis harbor a sparse but relatively characteristic microbial population.

Adolescent↗