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Immobilization of octadecyl ammonium chloride on the surface of titanium and its effect on microbial colonization in vitro.

The aim of our study was twofold: to immobilize an organosilicon quaternary ammonium salt (3-(trimethoxysilyl)-propyldimethyl-octadecyl ammonium chloride, Si-QAC) on the surface of pure titanium and to investigate the antimicrobial activity of Si-QAC-immobilized titanium against microbial adherence and biofilm formation. The results of ToF-SIMS analysis of Si-QAC-titanium suggested the possibility of immobilizing Si-QAC on titanium surface through Ti-O-Si coupling, and that Si-QAC treatment significantly reduced both the adherence and colonization of Candida albicans and Streptococcus mutans isolates. The antimicrobial activity was achieved through at least two mechanisms: the first was attributed to the octadecyl alkyl chain which inhibited initial adherence, and the second was attributed to the quaternary ammonium salt which killed initial adherent cells as well as retarded or inhibited subsequent microbial growth. Further, thermocycling did not significantly reduce the antimicrobial activity of Si-QAC-titanium, and no significant cytotoxicity of Si-QAC-titanium was observed in either cell viability test or proinflammatory cytokine production test using human gingival fibroblasts. These results, taken together, favorably suggested that Si-QAC treatment would be a helpful means to inhibit dental plaque or denture plaque formation.

Analysis of Variance↗

[The resistance of soil microorganisms to soil pollution by heavy metals].

Microbial communities of grey podzolized soil of Left-Bank Ukraine are characterized by high potential resistance to pollution with heavy metals: above 40% of organotrophic microorganisms are capable to resist to pollution with a mixture of heavy metals (Cu2+, Cd2+, Pb2+, Zn2+) in a dose of 20 MPC of each metal. Relative amount of resistant microorganisms in polluted soil increases up to 93%. White zeolites and biohumus are promising for reclamation of soils polluted with heavy metals. They decrease toxic effect of heavy metals on soil microflora and promote development of microflora resistant to pollution.

Colony Count, Microbial↗

Survival of microorganisms in smectite clays: implications for Martian exobiology.

Manned exploration of Mars may result in the contamination of that planet with terrestrial microbes, a situation requiring assessment of the survival potential of possible contaminating organisms. In this study, the survival of Bacillius subtilis, Azotobacter chroococcum, and the enteric bacteriophage MS2 was examined in clays representing terrestrial (Wyoming type montmorillonite) or Martian (Fe(3+)-montmorillonite) soils exposed to terrestrial and Martian environmental conditions of temperature and atmospheric pressure and composition, but not to UV flux or oxidizing conditions. Survival of bacteria was determined by standard plate counts and biochemical and physiological measurements over 112 days. Extractable lipid phosphate was used to measure microbial biomass, and the rate of 14C-acetate incorporation into microbial lipids was used to determine physiological activity. MS2 survival was assayed by plaque counts. Both bacterial types survived terrestrial or Martian conditions in Wyoming montmorillonite better than Martian conditions in Fe(3+)-montmorillonite. Decreased survival may have been caused by the lower pH of the Fe(3+)-montmorillonite compared to Wyoming montmorillonite. MS2 survived simulated Mars conditions better than the terrestrial environment, likely due to stabilization of the virus caused by the cold and dry conditions of the simulated Martian environment. The survival of MS2 in the simulated Martian environment is the first published indication that viruses may be able to survive in Martian type soils. This work may have implications for planetary protection for future Mars missions.

Azotobacter↗

Effect of nitrate and nitrite curing salts on microbial changes and sensory quality of non-fermented sausages.

The effects of nitrate and nitrite curing salts on microbial changes and sensory quality of non-fermented sausages of small diameter were investigated. During pre-ripening (day 5), levels of lactic acid bacteria and yeasts were slightly higher in nitrite-made sausages than in those made with nitrate. In contrast, nitrite discouraged the growth of psychrotrophs as occurs in fermented sausages. By the end of ripening (day 26), levels of microorganisms were similar in both batches of sausages except for psychrotrophs being higher in those made with nitrite. Nitrate-made sausages showed higher aroma and taste intensity.

Animals↗

Diversity, dynamics, and activity of bacterial communities during production of an artisanal Sicilian cheese as evaluated by 16S rRNA analysis.

The diversity and dynamics of the microbial communities during the manufacturing of Ragusano cheese, an artisanal cheese produced in Sicily (Italy), were investigated by a combination of classical and culture-independent approaches. The latter included PCR, reverse transcriptase-PCR (RT-PCR), and denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes (rDNA). Bacterial and Lactobacillus group-specific primers were used to amplify the V6 to V8 and V1 to V3 regions of the 16S rRNA gene, respectively. DGGE profiles from samples taken during cheese production indicated dramatic shifts in the microbial community structure. Cloning and sequencing of rDNA amplicons revealed that mesophilic lactic acid bacteria (LAB), including species of Leuconostoc, Lactococcus lactis, and Macrococcus caseolyticus were dominant in the raw milk, while Streptococcus thermophilus prevailed during lactic fermentation. Other thermophilic LAB, especially Lactobacillus delbrueckii and Lactobacillus fermentum, also flourished during ripening. Comparison of the rRNA-derived patterns obtained by RT-PCR to the rDNA DGGE patterns indicated a substantially different degree of metabolic activity for the microbial groups detected. Identification of cultivated LAB isolates by phenotypic characterization and 16S rDNA analysis indicated a variety of species, reflecting to a large extent the results obtained from the 16S rDNA clone libraries, with the significant exception of the Lactobacillus delbrueckii species, which dominated in the ripening cheese but was not detected by cultivation. The present molecular approaches combined with culture can effectively describe the complex ecosystem of natural fermented dairy products, giving useful information for starter culture design and preservation of artisanal fermented food technology.

Cheese↗

Reducing Salmonella on apples with wash practices commonly used by consumers.

The efficacy levels of practices used by consumers to wash smooth-surface fruits and vegetables were compared. Golden Delicious apples were spot inoculated near the blossom end with 50 microl of a cocktail of six serotypes of Salmonella enterica (with a total inoculum level of approximately 10(9) CFU per apple). The inoculum was dried for 1.5 h, and apples were either treated immediately or held for 24 h prior to treatment. Treatments included wetting with approximately 5 ml of water, vinegar (5% acidity), or a 200-ppm chlorine solution, rubbing for 5 or 30 s, rinsing with 200 to 600 ml of 24 or 43 degrees C water, and drying with a sterile paper towel. Residual populations of Salmonella were determined by rubbing the treated apple for 30 s in 20 ml of Dey-Engley neutralizing broth and plating on tryptic soy agar and bismuth sulfite agar. Rubbing treatments carried out for 5 and 30 s both resulted in a significant reduction in Salmonella populations (1 log10 CFU per apple) relative to populations on samples held for 30 s. A 5-s rub followed by a 200-ml flowing-water rinse reduced populations by 3 log10 CFU per apple. No further decrease in population was obtained by rinsing with 400 or 600 ml of water. Increasing the rinse water temperature to 43 degrees C did not significantly improve microbial removal. Drying the apple with a sterile paper towel resulted in an additional decrease of approximately 0.4 log10 CFU per apple. A reduction of 3.2 log10 CFU was achieved with a combination of wetting with water, rubbing for 5 s, rinsing with 200 ml of water, and drying with a paper towel for apples inoculated just prior to or 24 h before treatment. Reductions obtained for apples treated with 5% vinegar and with a 200-ppm chlorine solution were significantly larger (2.1 to 3.2 log10 CFU per apple, respectively) than those achieved with water.

Acetic Acid↗

PCR detection of Bacillus and Staphylococcus in various foods.

A broad-range PCR assay for the detection of bacteria belonging to Bacillus and Staphylococcus genera was developed. Primers targeting the bacterial 16S rRNA gene were newly designed and used in a PCR assay. To determine the specificity of the assay, 81 different bacterial strains (of 50 genera), 2 fungi, 3 animals, and 4 plants were tested. Results were positive for every tested Bacillus, Staphylococcus, or Aerococcus strain. In addition, the result for Listeria grayi was positive with lower PCR product. For all other bacterial strains and eukaryotes tested, results were negative. Bacterial DNA was prepared with the use of achromopeptidase and Chelex 100 resin from culture after growth in brain heart infusion medium. To test the sensitivity of this PCR assay for Bacillus or Staphylococcus genus, either Bacillus cereus or Staphylococcus aureus was inoculated into various foods with undetectable levels of endogenous microbial contamination as an indicator. Inoculation of bacteria at 10 to 30 CFU/g of food was followed by a 5-h enrichment culture step after which the PCR assay allowed the detection of bacterial cells. When the inoculation (B. cereus or S. aureus) of 10 to 90 CFU/g into noodle foods containing endogenous microflora (10(3) to 10(5) CFU/g) was followed by a 6-h enrichment culture step, the PCR assay detected the bacteria. Including the enrichment culture step, the entire PCR detection process can be completed within 8.5 h.

Bacillus↗

Effect of an essential oil-containing dentifrice on dental plaque microbial composition.

PURPOSE: To determine the effect of 6 months use of an essential oil-containing (EO) antiplaque/antigingivitis fluoride dentifrice on the balance of the oral microbial flora and on the emergence of resistant microbial forms by analysis of dental plaque and saliva. MATERIALS AND METHODS: The dentifrice essential oils consisted of a fixed combination of thymol, menthol, methyl salicylate, and eucalyptol. An identical fluoride-containing dentifrice without the essential oils served as the control. A subgroup of 66 subjects from a clinical trial population of 321 was randomly selected for characterization of their dental plaque microflora. Saliva was also cultured to monitor for the emergence of opportunistic pathogens. Supragingival plaque and saliva were harvested at baseline, after which subjects received a dental prophylaxis. Subjects were sampled again after 3 and 6 months of product use prior to clinical examination. Plaque was characterized for microbial content by phase contrast microscopy for recognizable cellular morphotypes and by cultivation on nonselective and selective culture media. Determination of the minimum inhibitory concentrations of the test agent against selected Actinomyces and Veillonella isolated bacterial species was conducted at all time points to monitor for the potential development of bacterial resistance. RESULTS: There were no statistically significant differences between the microbial flora obtained from subjects using the essential oil-containing dentifrice and the vehicle control for all parameters and time periods except for the percentage of spirochetes at 6 months and for percentage of "other" microorganisms at 3 months. The EO group exhibited a lower adjusted mean for both parameters. Additionally, there was no evidence of the development of bacterial resistance to the antimicrobial activity of the essential oils or the emergence of opportunistic pathogens.

Actinomyces↗

[Procedures for hand hygiene in German-speaking countries].

According to the field of application, strategies for the prevention of the transfer of microbial skin flora from the hands must consider the various categories of flora: transient, resident or stemming from infected lesions on the hands (infection flora). Depending on the species and virulence of the microorganism and of the susceptibility of the infection target, transient flora may or may not be of pathogenic importance. In contrast, resident skin flora is usually regarded as pathogenic only under certain circumstances such as in surgery, especially with transplantation of foreign bodies and in highly susceptible hosts. Microorganisms stemming from infected lesions are of proven pathogenicity. In the non-surgical field, only the transient and infection flora from the hands play a role. Such lesions are an absolute contraindication for patient-care, preparation of pharmaceuticals or foodstuff. In some procedures, the transmission of transient flora can be prevented by use of the non-touch technique ("instruments instead of fingers") or by the intelligent use of protective gloves. Hands already contaminated may be rendered safe by procedures for the elimination of transients such as handwashing, hygienic handwash and hygienic hand rub (in the order of increasing efficacy). Among all useable chemicals, ethanol, isopropanol and n-propanol (in the order of increasing efficacy) are the strongest and fastest agents. Furthermore, the duration of treatment (between 30 and 60 s) significantly influences the achievable reduction of microbial release. According to the new European standards (CEN) for testing chemical disinfectants and antiseptics, products for hygienic handwash must be significantly more efficacious than unmedicated soap, on artificially contaminated hands. In contrast, products for the hygienic hand rub must not be significantly less efficacious than a reference disinfection including isopropanol 60% vol rubbed onto the hands of the same volunteers during 1 min. By this, the average reduction of microbial release amounts to 4.2 to 4.4 lg, in our hands. The effectiveness of procedures for the hygienic handwash is usually significantly lower than that of alcoholic rubs. Therefore, in hospitals, they can be used only in certain indications such as patient care in reverse isolation, preparation of pharmaceuticals or foodstuff. In the surgical field, where not only transient but also resident flora is a cause of post-operative infection, the microbial release from the hands of the surgical team into the surgical wound must be prevented by using surgical gloves. Because of frequent glove lesions, a surgical hand disinfection is usually performed before donning gloves to keep a possible inoculumn as small as possible. Also in this field of application, alcoholic rubs proved to be significantly more effective than washing hands with antiseptic detergents. There exists a strong positive correlation of the reduction of microbial release and the duration of hand treatment, between 1 and 5 min. The European test standards (CEN) require products for surgical hand disinfection to be at least as efficacious as a reference disinfection of clean hands, which are constantly rubbed and kept wet with n-propanol 60% vol during 3 min. By this, the achievable average reduction of the microbial release ranges between 2.0 and 2.4 lg. In contrast, antiseptic washing procedures with preparations containing povidone-iodine, chlorhexidine gluconate or triclosan reduce the bacterial release within 2-5 min only by 0.5 to 1.2 lg. Some of them exert a bacteriostatic sustaining effect which is not found with alcoholic preparations. This, however, is not necessary with the latter as the initial bacterial reduction is that strong that restitution of the skin flora takes > 3 hours. Alcoholic preparations are at least as tolerable for the skin as antiseptic detergents, if not better, if they contain suitable emollients. Because dilution renders alcohols i

Colony Count, Microbial↗

Quality of fresh chicken breasts using a combination of modified atmosphere packaging and chlorine dioxide sachets.

The objective of this research was to observe the effect of chlorine dioxide (ClO2) combined with modified atmosphere packaging on the quality of fresh chicken breasts under refrigerated storage for 15 days. Each chicken breast was inoculated with a 4-log CFU/ml culture of Salmonella Typhimurium (nalidixic acid-resistant strain) and placed into a barrier foam tray. Fast- or slow-release ClO2 sachets were placed next to the chicken in each package. A control set of packages that did not contain a ClO2 sachet was also included in the study. Packages were flushed with either 100% N2 or 75% N2-25% CO2 and stored at 3 degrees C. Microbial analysis, CIE L.a.b. color, and sensory (appearance and aroma) were performed every 3 days for 15 days. Total plate counts for chicken increased steadily after 6 to 9 days of storage regardless of package atmosphere or ClO2 treatment. However, those treated with ClO2 sachets had 1 to 1.5 log CFU per chicken breast lower total plate counts compared with those without ClO2 sachets. After 15 days, samples treated with ClO2 (fast- and slow-release sachets) had significantly lower Salmonella Typhimurium (nalidixic acid-resistant strain) populations (approximately 1 log) compared with chicken that did not contain ClO2 sachets. The ClO2 adversely affected the color of the chicken in areas close to the sachet. No off-odor was detected by the sensory panelists.

Animals↗

Microbial translocation in neonates and infants receiving long-term parenteral nutrition.

OBJECTIVE: To explore whether episodes of endogenous septicemias due to microbial translocation are clinically relevant in neonates and infants who are receiving long-term parenteral nutrition (PN). DESIGN: Prospective observational cohort study of 2 years. SETTING: Neonates and infants who underwent surgical procedures and required PN because of gastrointestinal abnormalities. MEASUREMENTS: Surveillance cultures of the oropharynx and gut were obtained at the first of PN and thereafter twice each week. These cultures were processed for all microorganisms, except for coagulase-negative staphylococci, in a semiquantitative manner to detect overgrowth. A blood sample was taken for culture from both the central venous line and peripheral vein on clinical indication only. Microbial translocation was diagnosed when the microorganisms that were isolated from the blood sample were also carried in the throat and/or rectum within the 2 weeks preceding the episode of septicemia. MAIN RESULTS: Of 94 infants, 10 (11%) experienced 24 episodes of septicemia (ie, 7.3 septicemic episodes per 1000 days of PN). Six infants experienced 15 episodes of microbial translocation due to enteric microorganisms, including Escherichia coli, Klebsiella, Candida species, and enterococci. Microbial translocation occurred after a median of 58 days of PN (range, 32 to 286 days). The enteric organisms that caused septicemia were always present in the throat and/or rectum and in high concentrations ( > 10(5) colony-forming units per gram [ie, overgrowth]) in 60% of the translocation episodes. All but one episode occurred in infants with an abnormal serum bilirubin level ( > 17 mumol/L [0.99 mg/dl]). CONCLUSIONS: In neonates and infants who are receiving PN, septicemia may be a gut-related phenomenon.

Abdominal Muscles↗

Decontamination of strawberries using batch and continuous chlorine dioxide gas treatments.

Efficacy of chlorine dioxide (ClO2) gas in reducing Escherichia coli O157:H7 and Listeria monocytogenes on strawberries was determined using batch and continuous flow ClO2 gas treatment systems. Effects of continuous ClO2 gas treatment on total aerobic plate count, color, and residual ClO2 and chlorite on strawberries were also evaluated. Strawberries were spot inoculated with 7 to 8 log CFU per strawberry of each pathogen (E. coli O157:H7 and L. monocytogenes), stored for 1 day at 4 degrees C, and treated at 22 degrees C and 90 to 95% relative humidity with 0.2 to 4.0 mg/liter ClO2 gas for 15 or 30 min using a batch treatment system or with 0.6, 1.8, and 3.0 mg/liter for 10 min using a continuous treatment system. Surviving microbial populations were determined using a membrane-transfer plating recovery method. Increased ClO2 gas concentrations resulted in increased log reductions of each pathogen for both the batch and continuous systems. A batch treatment of strawberries with 4 mg/liter ClO2 for 30 min and continuous treatment with 3 mg/liter ClO2 for 10 min achieved greater than a 5-log reduction for both E. coli O157:H7 and L. monocytogenes. After continuous exposure to 3.0 mg/liter ClO2 gas for 10 min followed by 1 week of storage at 4 degrees C, no aerobic microorganisms were detected and the color of the strawberry surface did not change significantly (P > 0.05). Residues of ClO2 and chlorite on strawberries after the treatment were 0.19 +/- 0.33 mg ClO2 per kg and 1.17 +/- 2.02 mg Cl2 per kg, respectively, whereas after 1 week of storage no ClO2 residues were detected and residual chlorite levels were down to 0.07 +/- 0.12 mg Cl2 per kg. These results suggest that ClO2 gas treatment is an effective decontamination technique for improving the safety of strawberries while extending shelf life.

Chlorine Compounds↗

Eubacterial 16S-rDNA amplicon profiling: a rapid technique for comparison and differentiation of heterotrophic plate count communities from drinking water.

Determination of the heterotrophic plate count (HPC) is commonly used as a surrogate to assess the general microbial water quality in drinking water. For routine monitoring applications, the HPC is investigated in a quantitative way. However, qualitative data about the HPC bacterial community composition and/or population dynamics are required for particular situations. In order to provide fast and efficient qualitative approaches, molecular biological DNA profiling techniques seem to be suitable tools for the analysis of the total HPC community composition. In this work a DNA profiling technique is presented, which was recently demonstrated by our group to have potential for the rapid qualitative comparison and differentiation of HPC communities from raw and drinking water. The presented approach consists of a polymerase chain reaction (PCR)-based denaturing gradient gel electrophoresis (DGGE) for the generation of 16S-rDNA amplicon fingerprints from whole HPC community DNA extracts. In the context of this proceeding, the methodical background is presented and possible scientific merits as well as potential water management applications are discussed. Selected examples of (i) the demonstration of selective growth of HPC populations on different media and the comparison to the total in situ drinking water eubacterial community, (ii) the screening for HPC community variations at different locations of a drinking water distribution system, and (iii) the influence assessment on groundwater HPC communities by an infiltrating treated sewage effluent (bacterial source tracking) are given.

Colony Count, Microbial↗

The microbial content of non-sterile pharmaceuticals distributed in Norway.

Seventy-seven registered trademark pharmaceuticals and allied products, not required by the relevant monographs to comply with the test for sterility, were investigated for their microbial content. All the products examined complied with current regulations with respect to the numbers and types of microbes isolated, indicating the effectiveness of existing production practices in meeting existing standards. Gram-positive endospore-forming rods accounted for the majority of the bacteria isolated. Gram-negative rods were present for the most part in incidental numbers. However, some of these were of species that have been previously indicated as opportunistic pathogens and which should be considered as objectionable in pharmaceuticals. A number of the isolates showed antibiotic resistances unusual for the species in question.

Administration, Oral↗

Microbial controls on metal mobility under the low nutrient fluxes found throughout the subsurface.

Laboratory simulations and field studies of the shallow subsurface have shown that microbes and their extracellular products can influence the mobility of toxic metals from waste disposal sites. Modelling the transport of contaminants in groundwater may, therefore, require the input of microbial ecology data in addition to geochemical data, thus increasing the costs and the uncertainty of predictions. However, whether microbial effects on contaminant mobility occur extensively in the natural subsurface is unknown because the conditions under which they have been observed hitherto are generally unrepresentative of the average subsurface environment. Here, we show that microbial activity affects the mobility of a toxic trace metal (Cu) under the relatively low nutrient fluxes that dominate subsurface systems. More particularly, we show that under these low nutrient conditions, microbes and microbial products can immobilize metal but may themselves be subject to subsequent mobilization, thus complicating the pattern of metal storage and release. Our results show that the capability of microbes in the subsurface to change both the capacity of porous media to store metal, and the behaviour of metal that is released, is not restricted to the well researched environments close to sites of waste disposal. We anticipate our simulations will be a starting point for generating input data for transport models, and specifying the mechanism of metal remobilisation in environments more representative of the subsurface generally.

Colony Count, Microbial↗

[The homeostasis of the microbial communities in soils polluted by heavy metals].

Homeostatic potentialities of microbial communities in soils contaminated with heavy metals have been studied. To estimate the effect of contaminants on the soil microflora it was proposed to use the indices characterizing homeostasis of microbial community: microorganism survivability in contaminated soil and the period of restoration of their number, sensitivity to certain contaminants. In the model experiments on dark-grey podzolized soil it was shown that contamination with heavy metals (Cu2+, Cd2+, Sr2+, Pb2+, Hg2+) in the doses of 2 and 4 maximum permissible concentrations provokes a short-term inhibition of microorganisms development after which their number restores. Introduction of the mixture of metals evokes more intensive inhibition and extends the period of microorganism number restoration as compared with the effect of certain metals in the same doses.

Colony Count, Microbial↗

A quantitative microbiological study of bacterial vaginosis.

A quantitative microbial study of vaginal secretions from 100 women with bacterial vaginosis (BV) and 50 healthy women matched for age, was undertaken. This was aimed at comparing the bacterial flora colonising the vagina of sexually active asymptomatic women with those with BV and correlating bacterial counts with presence of symptoms. Of the total 475 bacteria isolated anaerobes were predominant among controls and among patients aerobes and anaerobes had equal prevalences. The increased viable counts of all anaerobes (> 10(7) cfu/g) were related with symptoms. Mobiluncus mulieris could be isolated from one patient and Chlamydia trachomatis antigen was absent in all samples tested. Bacteroides, G. vaginalis and Ureaplasma urealyticum were found to be the most commonly occurring group in women with BV. Lactobacillus, and probably Coryneforms have a protective role in health, preventing disease.

Adolescent↗

Two bacteria phylotypes are predominant in the Suiyo seamount hydrothermal plume.

Microbial diversity and populations in a hydrothermal plume that was present inside the caldera of the Suiyo Seamount, a submarine volcano on the Izu-Bonin Arc, were investigated by performing a phylogenetic analysis of the 16S rRNA gene and by using fluorescence in situ hybridization (FISH). Corresponding to transmissivity, an indicator of turbidity, the vertical total cell count as determined by 4',6'-diamidino-2-phenylindole (DAPI) staining varied from 5.6 x 10(4) to 1.1 x 10(5) cells ml(-1), and the apparent plume layer was assessed to be at a depth of 1,050 to 1,200 m inside the caldera and to contain 1.0 x 10(5) to 1.1 x 10(5) cells ml(-1). From microbial samples collected in the plume by an in situ filtration system, the following two major phylogenetic groups, which were closely related to sulfur-oxidizing microbes, were obtained: the SUP05 group belonging to the gamma subclass of the Proteobacteria (13 of 20 clones) and the SUP01 group belonging to the epsilon subclass of the Proteobacteria (5 of 20 clones). Specific oligonucleotide probes for these groups (SUP05-187 and SUP01-63) were designed and were used with various water samples obtained from the Suiyo Seamount. In the apparent plume layer, up to 66% of the total counts of microbial cells were estimated to be Bacteria cells that hybridized to EUB338, and few cells were identified by the archaeal probe ARCH915. Almost all Bacteria cells were hard to identify with the known group-specific probes, such as ALF19, GAM42a, and CF319, while 88 to 90% of the Bacteria cells hybridized with SUP05-187 and >98% of them were considered members of the SUP05 and SUP01 populations. In a low-temperature vent fluid emitted from a bivalve-colonized mound, the SUP05 cells accounted for >99% of the Bacteria cells, suggesting that a portion of the plume cells originated on the surface of the seafloor at a depth of about 1,380 m. From further analysis of cell morphology (i.e., cell size and cell elongation index) we inferred that the SUP05 cells were active in the plume layer at a depth of 1,050 to 1,200 m compared to the activity in a near-bottom layer, while many elongated cells were found between these layers. These findings suggest that the morphology and distribution of SUP05 cells have complex relationships with hydrothermal activities and water circulation. Although growth and production rates remain to be defined, we concluded that this Suiyo Seamount caldera has functioned as a natural continuous incubator for these two phylotypes of Bacteria in an aphotic deep-sea environment.

Colony Count, Microbial↗