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PubMedDate not supplied

[The count and distribution of saprophytic bacteria and bacteria of the E. coli group in the water of Odessa Bay and the adjacent waters].

The quantity and distribution of heterotrophic and enteric bacteria as indicators of the human activity influence on the marine ecosystem have been studied in water thickness of the Odessa Bay. It has been established that in the late autumn period the average quantity of heterotrophic bacteria in Bay waters is 6500 cells/ml and the number of the enteric ones--2400 cells/l. The spatial distribution of the heterotrophic bacterioplankton was mosaic and a tendency to decrease their quantity while moving away from the shore was not observed. This indicated the absence of the shore flow influence on the distribution of heterotrophic bacteria. On the contrary, the number of enteric bacteria significantly decreased with an increase of the distance from the shore, but on the greater part of the Bay they were absent at all. As to the vertical distribution of the heterotrophic bacteria quantity their maximum was in the water layer (0.5) below the surface, whereas the distribution of enteric bacteria did not correspond to this rule. Proceeding from the number of heterotrophic bacteria in the late autumn period waters of the Odessa Bay can be characterized as mesoeutrophic and according to the value of coli-index--as mesosaprobic ones.

Colony Count, Microbial↗
PubMed2002

Enumeration of total bacteria and bacteria with genes for proteolytic activity in pure cultures and in environmental samples by quantitative PCR mediated amplification.

Real-time quantitative PCR assays were developed for the absolute quantification of different groups of bacteria in pure cultures and in environmental samples. 16S rRNA genes were used as markers for eubacteria, and genes for extracellular peptidases were used as markers for potentially proteolytic bacteria. For the designed 16S rDNA TaqMan assay, specificity of the designed primer-probe combination for eubacteria, a high amplification efficiency over a wide range of starting copy numbers and a high reproducibility is demonstrated. Cell concentrations of Bacillus cereus, B. subtilis and Pseudomonas fluorescens in liquid culture were monitored by TaqMan-PCR using the 16S rDNA target sequence of Escherichia coli as external standard for quantification. Results agree with plate counts and microscopic counts of DAPI stained cells. The significance of 16S rRNA operon multiplicity to the quantification of bacteria is discussed.Furthermore, three sets of primer pair together with probe previously designed for targeting different classes of bacterial extracellular peptidases were tested for their suitability for TaqMan-PCR based quantification of proteolytic bacteria. Since high degeneracy of the probes did not allow accurate quantification, SybrGreen was used instead of molecular probes to visualize and quantify PCR products during PCR. The correlation between fluorescence and starting copy number was of the same high quality as for the 16S rDNA TaqMan assay for all the three peptidase gene classes. The detected amount of genes for neutral metallopeptidase of B. cereus, for subtilisin of B. subtilis and for alkaline metallopeptidase of P. fluorescens corresponded exactly to the numbers of bacteria investigated by the 16S rDNA targeting assay. The developed assays were applied for the quantification of bacteria in soil samples.

Bacillus cereus↗
PubMed1963

The fate of bacteria within phagocytic cells. I. The degradation of isotopically labeled bacteria by polymorphonuclear leucocytes and macrophages.

The intraleucocytic fate of a variety of P(32)- and C(14)-labeled bacteria has been studied in both polymorphonuclear leucocytes and macrophages. Both cell types brought about extensive degradation of bacterial lipids, nucleic acids, and proteins. Intracellular breakdown was primarily dependant upon the composition of the ingested particle rather than on the type or source of the phagocyte. Evidence is presented for the reincorporation of bacterial constituents into leucocyte lipid. More than 50 per cent of the acid-soluble degradation products of P(32)-labeled bacteria appear as inorganic phosphate. Bacterial RNA is degraded more readily than DNA. Following phagocytosis, labeled bacteria lose their pool of small molecular weight intermediates. This is followed by the degradation of acid-insoluble constituents. The majority of bacterial breakdown products are then excreted by the leucocyte and appear in the medium. Heat-killed bacteria were more readily broken down than viable organisms. Only small amounts of C(14)-labeled bacteria were completely oxidized by leucocytic enzymes to C(14)O(2). Acid extracts of polymorphonuclear leucocyte granules, which were highly bactericidal, liberated the acid-soluble constituents of labeled bacteria but did not significantly degrade bacterial macromolecules.

Bacteria↗
PubMed1992

Human bactericidal/permeability-increasing protein and a recombinant NH2-terminal fragment cause killing of serum-resistant gram-negative bacteria in whole blood and inhibit tumor necrosis factor release induced by the bacteria.

The bactericidal/permeability-increasing protein (BPI) of neutrophils and BPI fragments neutralize the effects of isolated Gram-negative bacterial lipopolysaccharides both in vitro and in vivo. Since endotoxin most commonly enters the host as constituents of invading Gram-negative bacteria, we raised the question: Can BPI and its bioactive fragments also protect against whole bacteria? To determine whether the bactericidal and endotoxin-neutralizing activities of BPI/fragments are expressed when Gram-negative bacteria are introduced to the complex environment of whole blood we examined the effects of added BPI and proteolytically prepared and recombinant NH2-terminal fragments on: (a) the fate of serum-resistant encapsulated Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa that survive the antibacterial actions of whole blood and (b) the ability of these bacteria to trigger cytokine release. Added BPI in nanomolar concentrations killed each of three encapsulated strains of E. coli and in closely parallel fashion inhibited tumor necrosis factor (TNF) release. Holo-BPI and its NH2-terminal fragment were equipotent toward a rough LPS chemotype K1-encapsulated strain, but the fragment was substantially more potent than holo-BPI toward two encapsulated smooth LPS chemotype strains. TNF release induced by K. pneumoniae and P. aeruginosa was also inhibited by both holo-BPI and fragment but, at the protein concentrations tested, P. aeruginosa was killed only by the fragment and K. pneumoniae was not killed by either protein. The bactericidal action of BPI/fragment toward E. coli is inhibited by C7-depleted serum, but accelerated by normal serum, indicating that BPI, acting in synergy with late complement components, enhances extracellular killing of serum-resistant bacteria. Thus, BPI and an even more potent NH2-terminal fragment may protect against Gram-negative bacteria in the host by blocking bacterial proliferation as well as endotoxin-mediated effects, not only as components of the intracellular antibacterial arsenal of the neutrophil, but also as potentially therapeutic extracellular agents.

Antimicrobial Cationic Peptides↗
PubMed2005

Monocyte-derived dendritic cells activated by bacteria or by bacteria-stimulated epithelial cells are functionally different.

Dendritic cells (DCs) are able to open the tight junctions between adjacent epithelial cells (ECs) and to take up both invasive and noninvasive bacteria directly from the intestinal lumen. In this study, we describe a tight cross talk between ECs and human monocyte-derived DCs (MoDCs) in bacterial handling across epithelial monolayers. We show that the release of proinflammatory mediators by ECs in response to bacteria is dependent on bacterial invasiveness and on the presence of flagella. This correlates with the capacity of EC-derived factors to modulate MoDC function. MoDCs incubated with supernatants of bacteria-treated ECs are "noninflammatory" as they release interleukin-10 (IL-10) but not IL-12 and can drive only T helper (Th)-2 type T cells. Moreover, noninflammatory MoDCs release chemokines aimed at recruiting Th2 and T-regulatory cells. In contrast, when MoDCs are incubated with ECs and bacteria in a transwell coculture system, and can contact directly the bacteria across stimulated EC monolayers, they are more inflammatory as they release IL-12 and IL-10 and induce both Th1 and Th2 responses. These results suggest that ECs are not simply a barrier to bacteria entering via the oral route, but they actively influence the activating properties of DCs.

Animals↗
PubMed1995

[Subgingival distribution of periodontopathic bacteria in periodontic patients and susceptibility of these bacteria to minocycline-HCl].

The present study was carried out to examine the distribution of six periodontopathic bacteria in deep periodontal pockets and to reconfirm the effect of Periocline on these periodontopathic bacteria. Samples from sixty-two periodontal pockets were collected at pocket depths of over 4 mm in twenty-one periodontitis patients aged 43 to 75 years. After sampling, Periocline was applied topically to the selected pockets once a week for four weeks and reexamined. The detected rates of the periodontopathic bacteria were Capnocytophaga sputigena (37.1%), Prevotella intermedia (22.6%), Porphyromonas gingivalis (22.6%), Fusobacterium nucleatum (20.1%), Actinobacillus actinomycetemcomitans (9.7%) and Eikenella corrodens (4.8%). The distribution of the bacteria was compound because two or three bacterial species were found to coexist. In view of the MIC of minocycline-HCI for these bacteria, increase of most of the measured bacteria was suppressed by the concentration of drugs, including Periocline. However, clinical strains of P. i. were considered to have low susceptibility to minocycline-HCl. In view of the effect of topical application of drugs, no significant differences were found. From these results, it was suggested that Periocline contained effective concentration of minocycline-HCl.

Adult↗
PubMed2006

Universal method for synthesis of artificial gel antibodies by the imprinting approach combined with a unique electrophoresis technique for detection of minute structural differences of proteins, viruses, and cells (bacteria). III: gel antibodies against cells (bacteria).

Artificial antibodies in the form of gel granules were synthesized from the monomers acrylamide and N,N'-methylenebisacrylamide by the imprinting method in the presence of Echerichia coli bacteria as template. The electrophoretic migration velocities of the gel antibodies (i) saturated with the antigen (Escherichia coli MRE-600), (ii) freed of the antigen, and (iii) resaturated with bacteria, were determinated by electrophoresis in a rotating narrow-bore tube of 245 mm length and the 2.5 and 9.6 mm inner and outer diameters, respectively. Removal of bacteria from the gel antibodies was made by treatment with enzymes, followed by washing with SDS and buffer. Gel granules becoming charged by adsorption of bacteria move in an electrical field. We obtained a significant selectivity of gel antibodies for E. coli MRE-600, since the granules did not interact with Lactococcus lactis; and when E. coli BL21 bacteria were added to the gels selective for E. coli MRE-600, a significant difference in the migration rate of the complexes formed with the two strains was observed indicating the ability of differentiation between the two strains. The gel antibodies can be used repeatedly. The new imprinting method for the synthesis of artificial gel antibodies against bioparticles described herein, and the classical electrophoretic analysis technique employed, thus represent - when combined - a new approach to distinguish between different types and strains of bacteria. The application area can certainly be extended to cover other classes of cells.

Acrylamide↗
PubMed2003

Influences of probiotic bacteria on organic acid production by pig caecal bacteria in vitro.

The mechanism of action of probiotics is largely unknown. A potential mechanism should be to increase the production of short-chain fatty acids (SCFA), known modulators of gut functions, by the bacterial ecosystem in the large intestine. The present paper reviews our recent studies in which the capacity of probiotic bacteria to increase the production of SCFA by pig caecal bacteria was investigated using batch-culture and continuous-culture techniques. All four commercial probiotic preparations and three strains of probiotic bacteria dose-dependently accelerated the net production of SCFA, succinic acid and lactic acid without changing the acid profile, and slowed the net production of NH4. Effects on organic acid production did not vary among different probiotic species. Neither probiotic preparations nor probiotic bacteria affected the organic acid production from glucose, gastric mucin, starch or lactose, or organic acids produced:added saccharide. Glucose abolished these effects of probiotic preparations. However, the capacity of probiotics to increase SCFA production was not modified by gastric mucin, starch or lactose. These results indicate that probiotic bacteria increase SCFA production by accelerating the breakdown of carbohydrates that are resistant to indigenous bacteria, and suggest that the concept of prebiotics in terms of SCFA production as a measure of probiotic function is arguable.

Ammonia↗
PubMed1995

Segmented filamentous bacteria are indigenous intestinal bacteria that activate intraepithelial lymphocytes and induce MHC class II molecules and fucosyl asialo GM1 glycolipids on the small intestinal epithelial cells in the ex-germ-free mouse.

In ex-germ-free mice conventionalized by association with fecal microorganisms, the induction fo major histocompatibility complex class II molecules and fucosylation of asialo GM1 glycolipid occur in the small intestinal epithelial cells (IEC). The intestinal intraepithelial lymphocytes (IEL), especially alpha beta T-cell receptor-bearing ones, also remarkably expand and show cytolytic activity. In this study, we investigated the immunological and physiological characteristics of the small intestine induced by a kind of indigenous bacteria of the small intestine, segmental filamentous bacteria (SFB), among chloroform-resistant intestinal bacteria. Monoassociation of SFB with germ-free mice was confirmed by the determination of the base sequences of polymerase chain reaction products of 16S rRNA genes of the fecal bacteria of these mice and in situ hybridization using fluorescein-labeled probes based on them. SFB increased the number of alpha beta TCR-bearing IEL and induced Thy-1 expression and cytolytic activity of IEL. The induction of MHC class II molecules and fucosyl asialo GM1 glycolipids and the increases in the mitotic activity and the ratio of the number of columnar cells to those of goblet cells also occurred in the small intestinal epithelial cells on monoassociation of these bacteria. SFB are important indigenous bacteria for the development of the mucosal architecture and immune system in the small intestine, at least in mice.

Animals↗
PubMed1982

Plants as sources of airborne bacteria, including ice nucleation-active bacteria.

Vertical wind shear and concentration gradients of viable, airborne bacteria were used to calculate the upward flux of viable cells above bare soil and canopies of several crops. Concentrations at soil or canopy height varied from 46 colony-forming units per m over young corn and wet soil to 663 colony-forming units per m over dry soil and 6,500 colony-forming units per m over a closed wheat canopy. In simultaneous samples, concentrations of viable bacteria in the air 10 m inside an alfalfa field were fourfold higher than those over a field with dry, bare soil immediately upwind. The upward flux of viable bacteria over alfalfa was three- to fourfold greater than over dry soil. Concentrations of ice nucleation-active bacteria were higher over plants than over soil. Thus, plant canopies may constitute a major source of bacteria, including ice nucleation-active bacteria, in the air.

Journal Article↗
PubMed1995

Total counts of marine bacteria include a large fraction of non-nucleoid-containing bacteria (ghosts).

Counts of heterotrophic bacteria in marine waters are usually in the order of 5 x 10(sup5) to 3 x 10(sup6) bacteria ml(sup-1). These numbers are derived from unspecific fluorescent staining techniques (J. E. Hobbie, R. J. Daley, and S. Jasper, Appl. Environ. Microbiol. 33:1225-1228, 1977; K. G. Porter and Y. S. Feig, Limnol. Oceanogr. 25:943-948, 1980) and are subsequently defined as total counts of bacteria. In samples from the Baltic Sea, the North Sea (Skagerrak), and the northeastern Mediterranean Sea, we found that only a minor fraction (2 to 32%) of total counts can be scored as bacteria with nucleoids. Lack of DNA no doubt means inactive cells; therefore, a much lower number of bacteria that grow at rates higher than those previously estimated must be responsible for the measured bacterial production in these seas. The remaining bacterium-sized and/or -shaped particles included in total counts may be cell residues of virus-lysed bacteria (ghosts) or remains of protozoan grazing.

Journal Article↗
PubMed2001

Effects of smoking and treatment status on periodontal bacteria: evidence that smoking influences control of periodontal bacteria at the mucosal surface of the gingival crevice.

BACKGROUND: We examined whether smoking status could influence growth of potentially pathogenic bacteria in the periodontal environment of treated and untreated periodontal patients. METHODS: We have previously reported effects of treatment status on marker bacteria in our patients. We established a history of any smoking during 6 months prior to microbiological sampling (F-ME, 16 smokers out of 64; MHM, 70 smokers out of 185). We used a commercial immunoassay to quantitate Porphyromonas gingivalis, Prevotella intermedia, and Actinobacillus actinomycetemcomitans in paper point samples from periodontal sites. RESULTS: Logistic regression showed that in smokers, neither P. gingivalis nor A. actinomycetemcomitans was quantitatively increased, while P intermedia was somewhat increased. Multiple regression demonstrated that smoking disrupts the positive relationship between increasing probing depth and increasing bacterial growth that is found in non-smokers. In smokers, growth of marker bacteria at shallow sites (< or =5 mm) was significantly increased to the levels found at deeper sites (>5 mm) in both smokers and non-smokers. Supragingival plaque biofilm was identified as a reservoir for marker bacteria; smokers and nonsmokers had equal ranges of oral cleanliness. CONCLUSIONS: Smoking-associated periodontitis is not simply a reflection of oral cleanliness. Smoking extends a favorable habitat for bacteria such as P. gingivalis, P. intermedia, and A. actinomycetemcomitans to shallow sites (< or =5 mm). Molecular byproducts of smoking interfere with mechanisms that normally contain growth of damaging bacteria at the surface of the oral mucosa in gingival crevices. In this way, smoking can promote early development of periodontal lesions.

Adult↗
PubMed2002

Microbial community structures of activated sludges dominated with polyphosphate-accumulating bacteria and glycogen-accumulating bacteria.

The microbial community structures of anaerobic-aerobic activated sludges, which are dominated with polyphosphate-accumulating bacteria (PABs) and glycogen-accumulating bacteria (GABs), were investigated by quinone profile method and dual staining DAPI (4',6-diamidino-2-phenylindol dihydrochloride) with fluorescent in situ hybridization (FISH). For both activated sludges, the predominant bacterial groups were the beta subclass of the Proteobacteria that contained mainly Q-8 as analyzed by the quinone profile method and gram-positive bacteria with a high DNA G+C content based on FISH. A 16S rRNA-targeted oligonucleotide probe (MP2) specific for M. phosphovorus, a known polyphosphate-accumulating bacterium, was also applied to PABs and GABs-enriched activated sludge to evaluate their possible role in the enhanced biological phosphorus removal (EBPR) process. Interestingly, the bacteria that hybridized with the probe specific to M. phosphovorus in the sludge dominated with PABs represent about 6.9% of the total microorganisms. In the sludge dominated with GABs, these bacteria were about 1.9%. These results suggest its possible involvement in the EBPR process.

Bacteria, Aerobic↗
PubMed1982

[Sensitivity of bacteria to chemotherapeutic agents (Zurich 1980). Part I. Bacteria isolated from hospital patients].

The frequency with which various bacteria are isolated from samples of hospitalized patients and the occurrence of sensitivity to antimicrobial agents in these bacteria are described. Data were collected and summarized by an electronic data processing system. Sensitivity data were obtained with the disc diffusion test carried out by the Bauer/Kirby method. E. coli and S. aureus were the most frequently isolated organisms. Pseudomonas sp. (in 90% Pseud. aeruginosa) took second place behind E. coli among gramnegative rod bacteria. As a rule more than 50% of grampositive and gramnegative bacteria were sensitive to betalactam antibiotics and broad spectrum agents. Aminocyclitols were the most potent drugs. Strains resistant to netilmicin or amikacin were only exceptionally observed. A comparison of the occurrence of sensitive bacteria from 1975 to 1980 revealed that with rare exceptions no decrease of sensitivity to standard drug occurred in E. coli, the Klebsiella-Enterobacter-Serratia group, Pseudo. aeruginosa and S. aureus.

Anti-Bacterial Agents↗
PubMed2004

A disintegration method for direct counting of bacteria in clay-dominated sediments: dissolving silicates and subsequent fluorescent staining of bacteria.

The masking of bacteria by abundant microparticles of the clay and silt fraction and cell losses due to sonication hampered direct enumeration of bacteria in sediments dominated by fine sediments. These problems can be circumvented by dissolving silicate fine particles using hydrofluoric acid and subsequent staining of bacteria by DTAF. The developed disintegration method partly replaces mechanical separation of bacteria from sediment particles by chemical disintegration of the silicates. Recovery efficiency ranged from 90% to 111% for different clays and clay-dominated sediments. Especially for the analysis of fine sediments and clays, this method circumvents both strong dilution of the sediment sample and harsh sonication. The method can also therefore be used in sediments where particle abundance is several orders of magnitude higher than bacterial abundance and simple dilution would not suffice in reliably counting bacteria.

Aluminum Silicates↗
PubMed1992

Efficacy of burning, tillage, and biocides in controlling bacteria released at field sites and effects on indigenous bacteria and fungi.

Decontamination treatments of burning and biocide application, alone and in combination with tillage, were evaluated for their ability to reduce populations of bacteria applied to the leaves of plants in field plots. In addition, the effects of these control methods on indigenous leaf and soil bacteria and fungi were assessed. Field plots of bush beans (Phaseolus vulgaris), sprayed with the bacterium Pseudomonas syringae, Pseudomonas fluorescens, or Erwinia herbicola, received the following treatments: (i) control, (ii) tillage, (iii) burning, (iv) burning plus tillage (burn-tillage), (v) Kocide (cupric hydroxide), (vi) Kocide plus tillage, (vii) Agri-Strep (streptomycin sulfate), and (viii) Agri-Strep plus tillage. Leaves and soil from the plots were sampled at 1 day before and at 1, 3, 7, 10, 14, 21, and 30 days after application of the decontamination treatments. The burn and burn-tillage treatments produced the most significant reductions in bacterial populations. The Agri-Strep treatment was more effective than the Kocide treatment in eliminating applied bacteria, but neither biocide produced consistent or persistent control. In contrast, the tillage treatment, alone or in combination with the Agri-Strep or Kocide treatments, had a short-term stimulatory effect and increased populations of applied bacteria and also levels of indigenous fungi and bacteria. Agri-Strep and Kocide treatments caused significant reductions in indigenous bacterial populations up to 14 days after application and in indigenous fungal populations on day 7 after application. Our results suggest that conventional plant disease control methods may not provide satisfactory control of genetically engineered microorganisms and indicate a need for further development of effective and selective methods to control release microorganisms at field sites.

Agriculture↗
PubMed1991

Competition for hydrogen by human faecal bacteria: evidence for the predominance of methane producing bacteria.

Studies of sludge have shown that some species of sulphate reducing bacteria outcompete methane producing bacteria for the common substrate H2. A similar competition may exist in human faeces where the methane (CH4) producing status of an individual depends on the faecal concentration of sulphate reducing bacteria. To determine if non-methanogenic faeces outcompete CH4 producing faeces for H2, aliquots of each type of faeces were incubated alone or mixed together, with or without addition of 10% H2 and/or 20 mmol/l sulphate. Methane producing faeces consumed H2 significantly more rapidly and reduced faecal H2 tension to a lower value compared with non-methanogenic faeces. The mixture of the two types of faeces yielded significantly more CH4 than CH4 producing faeces alone (mean (SD) 8.5 (1.3) v 2.9 (0.45) mmol/l of homogenate per 24 hours, p less than 0.01). Faecal sulphide concentrations were similar in CH4 producing and non-producing homogenates both before and after 24 hours of incubation. The addition of sulphate to the homogenates did not significantly influence CH4 production or sulphide formation. Our results suggest that in human faeces methane producing bacteria outcompete other H2 consuming bacteria for H2.

Bacteria, Anaerobic↗