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Mannose-binding activity of Escherichia coli: a determinant of attachment and ingestion of the bacteria by macrophages.

Recently, it was suggested that a mannose-specific lectin on the bacterial cell surface is responsible for the recognition by phagocytic cells of certain nonopsonized Escherichia coli strains. In this study we assessed the interaction of two strains of E. coli at different phases of growth with a monolayer of mouse peritoneal macrophages and developed a direct method with [(14)C]mannan to quantitate the bacterial mannose-binding activity. Normal-sized bacteria were obtained from logarithmic and stationary phases of growth. Nonseptated filamentous cells were formed by growing the organisms in the presence of cephalexin or at a restrictive temperature. Attachment to macrophages of all bacterial forms was inhibited by methyl alpha-d-mannoside and mannan but not by other sugars tested. The attachment of stationary phase and filamentous bacteria to macrophages, as well as their mannose-binding activity, was similar, whereas in the exponential-phase bacteria they were markedly reduced. The results show a linear relation between the two parameters (R = 0.98, P < 0.001). The internalization of the filamentous cells attached to macrophages during 45 min of incubation was much less efficient (20%) compared to that of exponential-phase, stationary-phase, or antibody-coated filamentous bacteria (90%). The results indicate that the mannose-binding activity of E. coli determines the recognition of the organisms by phagocytes. They further suggest that administration of beta-lactam antibiotics may impair elimination of certain pathogenic bacteria by inducing the formation of filaments which are inefficiently internalized by the host's phagocytic cells.

Animals↗

Microbial ecology of bulking and foaming activated sludge treating tannery wastewater.

This study involves a comprehensive survey of the microbial ecology of the activated sludge process treating tannery wastewater, emphasizing evaluation during periods of bulking and foaming. The survey also includes a detailed characterization of process influent and effluent. Observations always associate bulking with significant increase in protozoa species and rotifers, but decrease or disappearance of nematodes. Activated sludge sustained under normal conditions with a sludge volume index lower than 80 ml g(-1), exhibits all the properties of a compact and healthy zoogleal floc structure. Three types of filamentous bacteria prevail in the microscopic examinations. Two of the filamentous bacteria, Microthrix parvicella and Nostocoida limicola II, always appear as components of the floc structure under normal conditions and abundant or even excessive levels. The study identifies Gordona spp. as the main microorganism responsible for bulking and foaming, which starts by a Gordona contamination process inside the floc, later branching out and extending through adjacent flocs. When completed, this process increases sludge volume index values to over 200 ml g(-1). Microscopic examinations indicate significant morphological differences between filamentous microorganisms observed in this study and reported in the literature mostly for domestic sewage treatment.

Data Collection↗

Necrobacillosis in free-living male European bison in Poland.

In the Bialowieza Primeval Forest (Poland) a chronic disease of the external genital organs has been observed in free-living male European bison (Bison bonasus) since 1980. Investigations on this disease started in the late 1980s. The most striking findings are necrotic and ulcerative lesions of the prepuce and penis of bison aged from 6 mo to >10 yr. Histologic examination of tissue samples from the prepuce of six bison (9-mo- to 8-yr-old), and from the penis of two bison (3- and 8-yr-old), were characteristic of necrobacillosis. Masses of slender, Gram-negative, rod-like or filamentous bacteria occurred in necrotic tissue. At the periphery of necrotic tissue filamentous bacteria were often arranged in large clusters and strands that advanced towards healthy tissue. Immunolabeling and electron microscopy also suggest that these organisms are Fusobacterium sp.

Animals↗

Pathology of liver granulomas in turkeys.

Liver granulomas from five- and seven-week-old turkeys were studied by light and electron microscopy and by bacterial culture. Granulomas of five-week-old poults were composed of a caseous, necrotic center surrounded by consecutive zones of heterophils, giant cells, macrophages, and finally by lymphocytes. Tissue Gram and modified Dieterle stains demonstrated numerous gram-positive, filamentous bacteria in necrotic centers with radial extension of the bacteria to the zone of giant cells. Electron microscopic examination revealed septate, filamentous bacteria within an electron-dense milieu between heterophils, macrophages, and giant cells. An anaerobic, gram-positive bacillus and a staphylococcus were isolated from most granulomas examined. In broth, the bacillus formed long filaments or chains morphologically similar to those in tissue sections. The bacillus was identified tentatively as Eubacterium tortuosum. The staphylococcus had cultural reactions typical for Staphylococcus epidermidis. Intravenous inoculation of three-week-old poults with a mixture of E. tortuosum and S. epidermidis produced granulomas in liver and spleen and multiple mucosal ulcers in the duodenum and colon.

Animals↗

Bacteria Associated with the Gut Tract of Larval Stages of the Aquatic Cranefly Tipula abdominalis (Diptera; Tipulidae).

Scanning electron microscopy, light microscopy, and direct isolations were used to examine the distribution and diversity of bacteria in the gut tracts of larval stages of Tipula abdominalis. The animal had an enlarged hindgut which housed a diverse bacterial community in the lumen and directly attached to the gut wall. Distinct localization was noted, with the most dense and most diverse community anterior to the rectum. A distinct architecture of bacteria occurred in this region, characterized by a layering or a "weblike" array of filamentous bacteria overlying mats of bacteria closely associated with the gut wall. Although morphological diversity was high in the hindgut, filamentous bacteria were the dominant morphology observed. The attached microbiota, sloughed during ecdysis, recolonized to the same density and diversity observed before the molt. The majority of the isolatable bacterial types were facultatively anaerobic. The distinct localization and attached nature of the hindgut bacteria and the recolonization after each molt suggest they are indigenous to this region of the gut tract.

Journal Article↗

A new cytochrome subunit bound to the photosynthetic reaction center in the purple bacterium, Rhodovulum sulfidophilum.

The nucleotide sequence of the puf operon, which contains the genes encoding the B870 light-harvesting protein and the reaction center complex of the purple photosynthetic bacterium, Rhodovulum sulfidophilum, was determined. The operon, which consisted of six genes, pufQ, pufB, pufA, pufL, pufM, and pufC, is a new variety in photosynthetic bacteria in the sense that pufQ and pufC coexist. The amino acid sequence of the cytochrome subunit of the reaction center deduced from the pufC sequence revealed that this cytochrome contains only three possible heme-binding motifs; the heme-1-binding motif of the corresponding tetraheme cytochrome subunits was not present. This is the first exception of the "tetraheme" cytochrome family in purple bacteria and green filamentous bacteria. The pufC sequence also revealed that the sixth axial ligands to heme-1 and heme-2 irons were not present in the cytochrome either. This cytochrome was actually detected in membrane preparation as a 43-kDa protein and shown to associate functionally with the photosynthetic reaction center as the immediate electron donor to the photo-oxidized special pair of bacteriochlorophyll. This new cytochrome should be useful for studies on the role of each heme in the cytochrome subunit of the bacterial reaction center and the evolution of proteins in photosynthetic electron transfer systems.

Amino Acid Sequence↗

Genetic manipulation of carotenoid biosynthesis in the green sulfur bacterium Chlorobium tepidum.

The green sulfur bacterium Chlorobium tepidum is a strict anaerobe and an obligate photoautotroph. On the basis of sequence similarity with known enzymes or sequence motifs, nine open reading frames encoding putative enzymes of carotenoid biosynthesis were identified in the genome sequence of C. tepidum, and all nine genes were inactivated. Analysis of the carotenoid composition in the resulting mutants allowed the genes encoding the following six enzymes to be identified: phytoene synthase (crtB/CT1386), phytoene desaturase (crtP/CT0807), zeta-carotene desaturase (crtQ/CT1414), gamma-carotene desaturase (crtU/CT0323), carotenoid 1',2'-hydratase (crtC/CT0301), and carotenoid cis-trans isomerase (crtH/CT0649). Three mutants (CT0180, CT1357, and CT1416 mutants) did not exhibit a discernible phenotype. The carotenoid biosynthetic pathway in C. tepidum is similar to that in cyanobacteria and plants by converting phytoene into lycopene using two plant-like desaturases (CrtP and CrtQ) and a plant-like cis-trans isomerase (CrtH) and thus differs from the pathway known in all other bacteria. In contrast to the situation in cyanobacteria and plants, the construction of a crtB mutant completely lacking carotenoids demonstrates that carotenoids are not essential for photosynthetic growth of green sulfur bacteria. However, the bacteriochlorophyll a contents of mutants lacking colored carotenoids (crtB, crtP, and crtQ mutants) were decreased from that of the wild type, and these mutants exhibited a significant growth rate defect under all light intensities tested. Therefore, colored carotenoids may have both structural and photoprotection roles in green sulfur bacteria. The ability to manipulate the carotenoid composition so dramatically in C. tepidum offers excellent possibilities for studying the roles of carotenoids in the light-harvesting chlorosome antenna and iron-sulfur-type (photosystem I-like) reaction center. The phylogeny of carotenogenic enzymes in green sulfur bacteria and green filamentous bacteria is also discussed.

Alkyl and Aryl Transferases↗

Granulation in an upflow anaerobic sequencing batch reactor treating disintegrated waste activated sludge.

An upflow anaerobic reactor operated with a sequencing batch mode to enhance high rate digestion of raw and thermally disintegrated waste activated sludge with formation of granules. The gas production rate doubled when disintegrated waste activated sludge was introduced. Gradual granulation took place and the dispersed particles become coarse granulation as the operation continued. The granular sludge showed relatively higher specific methanogenic activity than the dispersed sludge. Bacterial morphology by a scanning electron microscope showed diversity of bacteria such as filamentous, rod and spherical shape in the section of granules. Filamentous bacteria, which might support the frame of a granule, were observed as long chains at the outer surface. Meanwhile, rod and spherical bacteria, which might play a role in the initial stage of granule formation, were observed from the inner surface of the granule. High rate digestion of sludge along with efficient liquid-solids separation was achieved due mainly to development of sludge granules within the upflow reactor.

Anaerobiosis↗

Molecular monitoring of bulking sludge in industrial wastewater treatment plants.

Fluorescent In Situ Hybridisation (FISH) was used to monitor the presence of filamentous microorganisms in industrial wastewater treatment plants (WWTPs). Monitoring with a restricted set of FISH probes in WWTPs from potato industry showed growth and decline of Thiothrix populations that could be linked to operational procedures. In a follow up project new FISH probes were developed for filamentous bacteria in industrial WWTPs and 70 WWTPs were analysed for presence of these filaments. Several newly described species of filamentous bacteria appear to be common and dominant in industrial WWTPs. Monitoring of a WWTP from textile industry showed growth and decline of one of these organisms when operational conditions in the plant were varied. The present paper demonstrates that bulking sludge in industrial wastewater treatment plants can effectively be monitored using a combination of standard chemical analyses and the FISH technique.

In Situ Hybridization, Fluorescence↗

Occurrence and control of filamentous bulking in aerated wastewater treatment plants of the French paper industry.

The occurrence of filamentous bacteria was investigated in 15 French pulp and paper activated sludge wastewater treatment plant (WWTP). Large filamentous populations were present in most of the plants. Identification carried out with conventional methods based on morphological features and staining techniques showed that the four main filamentous bacteria encountered in these industrial WWTP and responsible for bulking belong to the genera Thiothrix sp., Type 021 N, Haliscomenobacter hydrossis and Type 0092. During two years a specific survey was performed for three of these WWTP showing recurrent bulking phenomena. Data from WWTP performance, chemical data and filaments characterization were compared to correlate the presence of specific filaments with process operating conditions.

Bacteria↗

Filter clogging in coarse pore filtration activated sludge process under high MLSS concentration.

Coarse pore filtration activated sludge process is a type of hybrid process in which the secondary settling tank of the conventional activated sludge process is replaced by non- woven and coarse pore filter modules. The filter has pores, which are irregular in shape, and much bigger than micro-filtration membrane pores in size. The objective of the study is to find out the effect of the microbial community structure on filter clogging in the coarse pore filtration activated sludge process under high MLSS concentration in aerobic and anoxic/aerobic (A/A) conditions. Filter clogging started from day 65 and 70 in the A/A and aerobic process, respectively, but it was more severe in the A/A process compared to that in the aerobic process. EPS contents of sludge did not change significantly during the operation in both processes, and did not have a crucial effect on the observed filter clogging. There was no strong evidence for direct effect of the type and number of metazoa on filter clogging. The main difference between aerobic sludge and A/A sludge during the filter clogging period was the relative abundance of filamentous bacteria. According to the obtained results, it can be concluded that a higher presence of filamentous bacteria could reduce the severity of filter clogging in a coarse pore filtration activated sludge process.

Bacteria, Aerobic↗

Centrin-like filaments in the cytopharyngeal apparatus of the ciliates Nassula and Furgasonia: evidence for a relationship with microtubular structures.

The cytopharyngeal apparatus in the Nassulinid ciliates Nassula and Furgasonia is a highly specialized microtubular/filamentous organelle designed for ingestion of organisms such as filamentous bacteria. From studies on living cells, it was previously shown that this organelle, also called "feeding basket," guides the filamentous bacteria and manipulates them to some extent during the early steps of ingestion. This results in a complex sequence of movements where the basket is successively dilated and constricted in its upper part. Whereas some of these movements (dilation) seem to be intrinsic to the microtubular components of the basket, others (constriction) are believed to be mediated by contractile filamentous structures [Tucker, 1968: J. Cell Sci. 3:493-514]. In this study, we have used antibodies raised against ciliate centrins to demonstrate these proteins by Western blot and immunocytochemical methods in Nassula and Furgasonia. In both ciliates, a 20-kDa centrin immunoanalog was localized in the upper (contractile) part of the cytopharyngeal apparatus. Immunoelectron microscopy revealed that cytopharyngeal centrin is engaged in filamentous material, forming a sphincter-like structure possibly involved in the movements of contraction. Interestingly, physical links were noted between filaments labeled for centrin and cytopharyngeal microtubules. The mechanistic implications of these findings are discussed.

Animals↗

Influence of transient substrate overloads on the proliferation of filamentous bacterial populations in an activated sludge pilot plant.

Using oligonucleotide probes directed at the rRNA of filamentous bacteria, this study looks at the influence of the components of transient substrate overloads on the growth of the dominant filamentous bacteria of activated sludge fed by a synthetic substrate. By dissociating the massive input of organic matter from the oxygen shortage that the latter generally induces, it is revealed that each of these factors applied alone, induces only transitory, small-scale growth of the filaments Nostocoida limicola, Haliscomenobacter hydrossis. Thiothrix and of type 021N. In contrast, combining them during a reconstituted transient substrate overload with an artificially created oxygen deficit, induces very fast growth of H. hydrossis which is responsible for establishing major proliferation. This massive proliferation was easily reduced by chlorination.

Bacteria↗

Linking floc structure and settling properties to activated sludge population dynamics in an SBR.

Over a period of 227 days properties of activated sludge grown in an sequencing batch reactor (SBR) operated under stable conditions were analyzed. Settling properties (sludge volume index (SVI)) of the activated sludge were compared with on-line measurements of floc size and size distribution obtained by using a laser light scattering technique (Malvern Mastersizer/S, Malvern, UK), and with measurements of microbial community dynamics analyzed by denaturing gradient gel electrophoresis (DGGE) patterns of 16S rRNA genes. In addition, microscopical observations were used to confirm the results. Three distinct stages in the SBR evolution were observed. In the first stage the structural floc properties showed predominant presence of floc-forming bacteria in the activated sludge. A good correlation between floc size, properties and microbial community evolution was observed. The second stage showed a good balance between floc-forming and filamentous bacteria, with good settling properties and a highly dynamic community in the SBR. In the third stage, an increase in the filamentous bacteria, which became predominant in the system was observed. Again, a good correlation between settling properties and floc size distribution was obtained and a new dominant species was observed in the DGGE patterns, which can be assumed to be a filamentous organism.

Bioreactors↗

Permanent presence of grazing-resistant bacteria in a hypertrophic lake.

The size structure of planktonic bacteria from a hypertrophic lake was investigated at 5- to 15-day intervals by means of a semiautomatic image analysis system during 1 year. Characteristic of this bacterial assemblage was the permanent presence of large filamentous bacteria and small cocci with cell sizes of <0.01 (mu)m(sup3). These filamentous bacteria, sometimes longer than 200 (mu)m and with cell volumes of up to 276 (mu)m(sup3), are larger than nanoflagellates (<20 (mu)m) and, even, metazoans living in the lake. Although they account for only 4 to 16% of bacterial abundance, their contribution to total bacterial biovolume was between 45 and 86%. An analysis of the food web structure indicates that this particular bacterial size structure may be the consequence of a strong bacterivory pressure by nanoflagellates and the absence of other larger bacterivores. The persistence of bacterial forms resistant to grazing has important consequences for the carbon flow within the microbial food web.

Journal Article↗

Biodegradation of aromatic compounds and TCE by a filamentous bacteria-dominated consortium.

The Michaelis-Menten biodegradation kinetics (k and Ks) of aromatic compounds and trichloroethene (TCE) by an aerobic enrichment culture grown on phenol and dominated by a unique filamentous bacterium were measured. The average k and Ks values for phenol, benzene (B), toluene (T), ethylbenzene (E), o-xylene (oX), p-xylene (pX), naphthalene and TCE in g per g VSS-d and mg L-1 were 5.72 and 0.34, 1.20 and 0.51, 2.09 and 0.47, 0.77 and 0.23, 0.61 and 0.16, 0.73 and 0.23, 0.17 and 0.18, and 0.16 and 0.18, respectively. Significant variability in these measured kinetics was noted between tests conducted over the 5-month period during which the fed-batch culture with a 5-day solids retention time was maintained; the coefficient of variation of the k and Ks values ranged from 11-43% and 4-50%, respectively. This variation was significantly greater than the method measurement error on a given date. Degradation of BTEoXpX mixtures could be described by a basic competitive inhibition model. Batch tests during which the culture was fed individual BTEX compounds showed the culture grew poorly on the xylenes and had poor subsequent xylene degradation rates. This work indicates the potential to simultaneously treat a mixture of volatile organic compounds using this consortium, and the ability to predict the mixture biodegradation rates on the basis of the individual compound biodegradation kinetics.

Algorithms↗

Ecophysiology of the filamentous Alphaproteobacterium Meganema perideroedes in activated sludge.

A comprehensive study of the ecophysiology of the filamentous Meganema perideroedes affiliated to the Alphaproteobacteria, possessing a "Nostocoida limicola Type II" filamentous morphology was conducted. This morphotype often causes serious bulking problems in activated sludge wastewater treatment plants, and hardly anything is known about its physiology. The study was carried out by applying a suite of in situ methods in an industrial activated sludge treatment plant with excessive growth of this species. The experiments revealed a very versatile organism able to take up a large variety of organic substrates under aerobic conditions. It had a remarkably high storage capacity forming polyhydroxyalkanoates from most substrates tested. When nitrate was present as e-acceptor, the number of substrates to be consumed by M. perideroedes was more restricted compared to aerobic conditions. With nitrite as e-acceptor, only acetate and glucose among the substrates tested could be assimilated and used for storage and possibly growth. This indicated that M. perideroedes might be able to denitrify under certain conditions, which is unusual for filamentous bacteria in activated sludge. No substrate uptake or storage was seen under anaerobic conditions. M. perideroedes was relatively hydrophobic, compared to other filamentous bacteria and microcolonies present in the sludge, indicating the presence of a hydrophobic sheath. Several excreted surface-associated exoenzymes were detected in the sludge, but M. perideroedes never showed any activity, except once after a breakdown in the production facility. This confirmed that M. perideroedes mainly grows on soluble substrates. Based on the studies of the ecophysiology of M. perideroedes, potential control strategies are suggested.

Aerobiosis↗

Application of the Fluorescent-Antibody Technique for the Detection of Sphaerotilus natans in Activated Sludge.

Sphaerotilus natans, one of the most widely reported causes of bulking in activated sludge, can exist both within and outside of a sheath. It can easily be confused with similar activated sludge bacteria and thus can be overlooked when present in low numbers. Fluorescent antiserum was successfully prepared against the nonfilamentous form and was shown to be highly specific, showing no reaction with either pure cultures of similar filamentous bacteria or entirely unrelated organisms. It did, however, show a lack of strain specificity since it reacted with S. natans isolates from the Federal Republic of Germany and the United States and with filamentous bacteria in South African activated sludges. Fluorescent antibody is capable of penetrating the filaments of S. natans to stain the cells individually. The use of fluorescent antiserum in the identification of S. natans filaments obscured by activated sludge flocs and other suspended matter was simple since the cells stained brightly and could be observed through the less dense matter, while the use of other microscope techniques would be hampered by these obstructions. The use of fluorescent antibody will facilitate ecological studies of S. natans in activated sludge and other aqueous environments.

Journal Article↗