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PubMed1999

Responses of the soil microbiota to elevated CO2 in an artificial tropical ecosystem.

Plants in artificial tropical ecosystems were grown under ambient (340 microl l(-1)) and elevated (610 microl l(-1)) atmospheric CO2 for 530 d under low-nutrient conditions on a substrate free of organic C. At the end of the experiment a number of soil chemical and microbiological variables were determined. Although we found no changes in total soil organic matter under elevated CO2, we did find that after physical fractionation the amount of organic C in the supernatant (< 0.2 microm) and the amount of water extractable organic C (WEOC) was lower under elevated CO2. The extractable optical density (OD) indicated a higher degree of humification for the elevated than for the ambient CO2 samples (P = 0.032). Microbial biomass C was not significantly altered under high CO2, but total bacterial counts were significantly higher. The microbial biomass C-to-N ratio was also higher at elevated (15.0) than at ambient CO2 (10.0). The number of mycorrhizal spores was lower at high CO2, but ergosterol contents and fungal hyphal lengths were not significantly affected. Changes were found neither in community level physiological profiles (CLPPs) nor in the structural attributes (phospholipid fatty acids, PLFAs) of the microbial community. Overall, the effects on the soil microbiota were small, perhaps as a result of the low nutrient supply and low organic matter content of the soil used in our study. The few significant results showing changes in specific, though relatively minor, organic matter pools may point to possible long-term changes of the more major pools. Furthermore, the data suggest increased competition between plants and microbes for N at high CO2.

Bacterial Physiological Phenomena↗

PubMed2005

Survival, growth and persistence under farm conditions of a Lactobacillus plantarum strain inoculated into liquid pig feed.

AIMS: To investigate the survival and persistence of Lactobacillus plantarum REB1 in the fermentation process of liquid pig feed on a working farm in standard production conditions. METHODS AND RESULTS: Two feed types, a control diet [nonfermented liquid feed (NFLF)] and a fermented diet [fermented liquid feed (FLF)], were compared. A rifampicin-resistant mutant L. plantarum REB1-Rif was used to initiate the fermentation of the feed. Inoculation with the experimental strain was repeated one or two times per week throughout the three month growing period. Four microbial groups were followed using standard microbiological techniques, as well as pH. Lactic acid bacteria (LAB) counts were high already at the beginning in FLF, while it took nine days to reach the corresponding LAB levels in NFLF. Yeasts were stable in FLF, whereas in NFLF there were occasional high counts during the first week. The numbers of Enterobacteriaceae were low, although in NFLF they were variable. The average pH in NFLF was 4.5 and 4 in FLF. CONCLUSIONS: The inoculation of L. plantarum REB1-Rif provided a LAB population of log 9 colony forming units (CFU) ml(-1) from the first feeding day, stable numbers of yeast and pH, and a drastic reduction of Enterobacteriaceae. SIGNIFICANCE AND IMPACT OF THE STUDY: The inoculation by L. plantarum REB1-Rif offered a FLF microbiologically stable from the first week in actual production conditions.

Animal Feed↗

PubMed2003

Physiological and community responses of established grassland bacterial populations to water stress.

The effects of water stress upon the diversity and culturable activity of bacterial communities in the rhizosphere of an established upland grassland soil have been investigated. Intact monoliths were subjected to different watering regimens over a 2-month period to study community adaptation to moisture limitation and subsequent response to stress alleviation following rewetting. Genetic diversity was analyzed with 16S-based denaturing gradient gel electrophoresis (DGGE) of total soil-extracted DNA (rRNA genes) and RNA (rRNA transcripts) in an attempt to discriminate between total and active communities. Physiological response was monitored by plate counts, total counts, and BIOLOG-GN2 substrate utilization analyses. Controlled soil drying decreased the total number of CFU on all the media tested and also decreased the substrate utilization response. Following rewetting of dried soil, culture-based analyses indicated physiological recovery of the microbial population by the end of the experiment. In contrast, DGGE analyses of community 16S rRNA genes, rRNA transcripts and cultured communities did not reveal any changes relating to the moisture regimens, despite the observed physiological effects. We conclude that the imposed moisture regimen modulated the physiological status of the bacterial community and that bacterial communities in this soil are resistant to water stress. Further, we highlight the need for a reexamination of rRNA transcript-based molecular profiling techniques as a means of describing the active component of soil bacterial communities.

Bacteria↗

PubMed1990

Analysis of positive cultures from endodontically treated teeth: a retrospective study.

A study of 12,150 endodontically treated teeth from 11,797 patients in a private patient setup revealed an incidence of 10.2% positive cultures obtained just before obturation of the root canal. Streptococcal predominance was reflected by a presence of nearly 60% in the pure culture and nearly 98% in the mixed cultures. No significant changes in microbial predominance or bacterial sensitivities were detected over the 7-year period of study. Ampicillin, cephalothin, erythromycin, and penicillin were the most effective antibiotics against the viridans streptococci, the most prevalent organism in the infected teeth.

Adolescent↗

PubMed2001

Phenotypic and molecular identification and clustering of lactic acid bacteria and yeasts from wheat (species Triticum durum and Triticum aestivum) sourdoughs of Southern Italy.

The microflora of 25 wheat sourdoughs from the Apulia region, Southern Italy, was characterized. The sourdoughs were mainly produced from Triticum durum wheat. The number of lactic acid bacteria and yeasts ranged from ca. log 7.5 to log 9.3 colony forming units (cfu)/g and from log 5.5 to log 8.4 cfu/g, respectively. About 38% of the 317 isolates of lactic acid bacteria were identified by conventional physiological and biochemical tests. Phenotypic identification was confirmed by 16S rDNA and 16S/23S rRNA spacer region PCR. Overall, 30% of the isolates were identified as Lactobacillus sanfranciscensis, 20% as Lb. alimentarius, 14% as Lb. brevis, 12% as Leuconostoc citreum, 7% as Lb. plantarum, 6% as Lactococcus lactis subsp. lactis, 4% as Lb. fermentum and Lb. acidophilus, 2% as Weissella confusa and 1% as Lb. delbrueckii subsp. delbrueckii. Some of these species have not been previously isolated from sourdoughs. Since bakers yeast is widely used in sourdough production, Saccharomyces cerevisiae was largely found. The phenotypical relationships within the main lactic acid bacteria identified were established by using cluster analysis. A microbial map of the 25 sourdoughs was plotted showing characteristic associations among lactic acid bacteria and differences in the lactic acid bacteria species which were mainly due to the species of wheat flour, use of bakers yeast and type of bread.

Bread↗

PubMed1992

Enumeration of subgingival species on primary isolation plates using colony lifts.

This study evaluated the feasibility of using a colony lift method and DNA probes to enumerate bacterial species cultured on primary isolation plates. Fourteen digoxigenin-labeled whole chromosomal DNA probes representing 12 subgingival species were validated by hybridization with colony lifts prepared from 249 reference strains of 51 species grown on Trypticase soy agar plates supplemented with 5% sheep blood. Colonies of reference strains were lifted onto Nytran filters from plates and treated to lyse cells, remove cellular proteins, denature and fix microbial DNA to the filters. Positive reactions were detected with an anti-digoxigenin antibody conjugated to alkaline phosphatase and revealed by bromo-chloro-indolyl phosphate and nitroblue tetrazolium. Cross-reactions were not observed for 13/14 probes, but 2 strains of Streptococcus mitis reacted with the probe to Streptococcus sanguis II. Subgingival plaque samples were taken by means of a sterile curette from mesiobuccal surfaces of teeth present in each of 26 subjects with differing periodontal disease states. Samples were dispersed, diluted, plated and incubated anaerobically for 7 d at 35 degrees C. Colonies were lifted as described above. Filters were cut into sections and hybridized with the 14 digoxigenin-labeled DNA probes. The probes were used to enumerate the test species and the total number of isolates was determined in 711 plaque samples. The colony lift method and DNA probes provided a sensitive, economical and quantitative method for enumerating cultivable microbial species in subgingival plaque samples. In addition, the amplification provided by growing the organisms on agar plates facilitated determination of numbers of organisms in small plaque samples, such as those from healthy sites.

Actinomyces↗

PubMed2002

Ongoing modification of Mediterranean Pleistocene sapropels mediated by prokaryotes.

Late Pleistocene organic-rich sediments (sapropels) from the eastern Mediterranean Sea harbor unknown, metabolically active chemoorganotrophic prokaryotes. As compared to the carbon-lean intermediate layers, sapropels exhibit elevated cell numbers, increased activities of hydrolytic exoenzymes, and increased anaerobic glucose degradation rates, suggesting that microbial carbon substrates originate from sapropel layers up to 217,000 years old. 16S ribosomal RNA gene analyses revealed that as-yet-uncultured green nonsulfur bacteria constitute up to 70% of the total microbial biomass. Crenarchaeota constitute a smaller fraction (on average, 16%). A slow but significant turnover of glucose could be detected. Apparently, sapropels are still altered by the metabolic activity of green nonsulfur bacteria and crenarchaeota.

Alkaline Phosphatase↗

PubMedDate not supplied

[The microbial cenosis of the tonsils in healthy children and in those frequently ill].

The microbial cenosis of tonsils has been studied in healthy and ailing children. It has been found out that Staphylococcus aureus and S. epidermidis, beta-hemolytic streptococcus, rarely gram-negative flora and candidae were most often isolated in the ailing children. Normal microflora of healthy children mainly included lactic acid bacteria whose level considerably decreased in ailing children that is most distinctly expressed in children with decompensated tonsillitis. To correct the disturbances in microbial cenosis during complex therapy we used lactobacteria to treat the nasopharynx with the suspension of these bacteria. Positive clinical effect with following positive results of bacteriological investigation has been obtained in all patients.

Adolescent↗

PubMed2005

Hydrolytic activity of alpha-amylase in anaerobic digested sludge.

Hydrolysis is usually considered to be a rate-limiting step in anaerobic digestion. For improving anaerobic solid waste treatments, it is essential to elucidate the mechanism of hydrolysis. In this study, alpha-amylase, one of the hydrolytic enzymes, was investigated for the elucidation of more precise mechanism of hydrolysis. Alpha-amylase activity of solid starch-degrading bacteria (SDB) was estimated through batch experiments with several different substrates and with distinction between cell-bound and cell-free alpha-amylase. Monitoring of newly isolated strains of SDB was done by fluorescence in situ hybridization. Results indicated that cell-bound alpha-amylase is chiefly responsible for the hydrolysis in the digested sludge, providing very useful information that the contact between microbial cells and solids is significantly important. The activity of alpha-amylase of the digested sludge remained quite low when not required, but increased as they recognized appropriate substrates. Several-fold higher activity was obtained for starch or maltose as compared to glucose only.

Aeromonas↗

PubMed1998

Quality of commercial shelf-stable soymilk products.

Four brands of shelf-stable soymilks (three lots each) were evaluated for their quality. All samples were tested for trypsin inhibitor activity, microbial load (coliforms, aerobic mesophiles, and psychrotrophs), solids content, pH, color, and viscosity. Storage stability at 5 degrees C of the opened original containers was evaluated by microbiological analyses. Physical properties were consistent among three lots of the same brand, but varied among brands. Trypsin inhibitor activity ranged from 3.43 to 10.35 mg/g dried soymilk. The pH values of the soymilks varied between 6.41 and 7.34. Viscosity was measured between 16.5 and 26.4 mPa. Solids content varied considerably among soymilks, from 6.88 to 12.37% (wt/vol). Freshly opened commercial soymilk contained fewer than one microorganism per ml in all microbial counts. During storage at 5 degrees C, microbial counts increased sharply after 2 to 3 weeks. A single microorganism predominated in each stored sample. Shelf life (defined as the time during which aerobic mesophile counts remained below 10(6) CFU/ml) varied from 3 to 4 weeks. The appearance and pH of soymilks remained unchanged even after 4 weeks of storage. Five types of microorganisms were found in stored soymilk samples; none of them produced acid.

Bacteria↗

PubMed2000

An evaluation of sampling and laboratory procedures for determination of heterotrophic plate counts in dental unit waterlines.

BACKGROUND: The high numbers of heterotrophic microorganisms that have been cultured from dental unit waterlines (DUWs) have raised concern that this water may exceed suggested limits for heterotrophic plate counts (HPCs). The main purpose of this investigation was to examine HPC variability in DUWs and to examine in detail the effect of laboratory processing of water samples on HPC values. METHODS: Water samples were collected from dental offices either at the beginning of or during the clinic day and were transported to the laboratory, where they were analyzed. RESULTS: Measuring HPC levels within an office would involve testing all units, because significant differences were found between units connected to the same municipal water supply. Within a unit, the average microbial count from high-speed lines was approximately twice the average count from air/water lines. The laboratory processing of water samples significantly affected the numbers of heterotrophic microorganisms that were recovered. Incubation temperature, time and media, as well as neutralization of residual chlorine, all had significant effects on the HPC values. However, no significant differences in microbial counts were found between samples plated with the spread plate method on R2A agar and those plated with the pour plate method with Plate Count Agar. CONCLUSIONS: Dental organizations have suggested target limits in terms of numbers of heterotrophic microorganisms recovered in water from dental units, but standards for laboratory handling must be established as well. A protocol for sample collection and laboratory handling is proposed.

Bacteriological Techniques↗

PubMed2006

Size and structure of bacterial, fungal and nematode communities along an Antarctic environmental gradient.

The unusually harsh environmental conditions of terrestrial Antarctic habitats result in ecosystems with simplified trophic structures, where microbial processes are especially dominant as drivers of soil-borne nutrient cycling. We examined soil-borne Antarctic communities (bacteria, fungi and nematodes) at five locations along a southern latitudinal gradient from the Falkland Islands (51 degrees S) to the base of the Antarctic Peninsula (72 degrees S), and compared principally vegetated vs. fell-field locations at three of these sites. Results of molecular (denaturing gradient gel electrophoresis, real-time PCR), biochemical (ergosterol, phospholipid fatty acids) and traditional microbiological (temperature- and medium-related CFU) analyses were related to key soil and environmental properties. Microbial abundance generally showed a significant positive relationship with vegetation and vegetation-associated soil factors (e.g. water content, organic C, total N). Microbial community structure was mainly related to latitude or location and latitude-dependent factors (e.g. mean temperature, NO3, pH). Furthermore, strong interactions between vegetation cover and location were observed, with the effects of vegetation cover being most pronounced in more extreme sites. These results provide insight into the main drivers of microbial community size and structure across a range of terrestrial Antarctic and sub-Antarctic habitats, potentially serving as a useful baseline to study the impact of predicted global warming on these unique and pristine ecosystems.

Animals↗

PubMed2000

Relative contributions of bacteria, protozoa, and fungi to in vitro degradation of orchard grass cell walls and their interactions.

To assess the relative contributions of microbial groups (bacteria, protozoa, and fungi) in rumen fluids to the overall process of plant cell wall digestion in the rumen, representatives of these groups were selected by physical and chemical treatments of whole rumen fluid and used to construct an artificial rumen ecosystem. Physical treatments involved homogenization, centrifugation, filtration, and heat sterilization. Chemical treatments involved the addition of antibiotics and various chemicals to rumen fluid. To evaluate the potential activity and relative contribution to degradation of cell walls by specific microbial groups, the following fractions were prepared: a positive system (whole ruminal fluid), a bacterial (B) system, a protozoal (P) system, a fungal (F) system, and a negative system (cell-free rumen fluid). To assess the interactions between specific microbial fractions, mixed cultures (B+P, B+F, and P+F systems) were also assigned. Patterns of degradation due to the various treatments resulted in three distinct groups of data based on the degradation rate of cell wall material and on cell wall-degrading enzyme activities. The order of degradation was as follows: positive and F systems > B system > negative and P systems. Therefore, fungal activity was responsible for most of the cell wall degradation. Cell wall degradation by the anaerobic bacterial fraction was significantly less than by the fungal fraction, and the protozoal fraction failed to grow under the conditions used. In general, in the mixed culture systems the coculture systems demonstrated a decrease in cellulolysis compared with that of the monoculture systems. When one microbial fraction was associated with another microbial fraction, two types of results were obtained. The protozoal fraction inhibited cellulolysis of cell wall material by both the bacterial and the fungal fractions, while in the coculture between the bacterial fraction and the fungal fraction a synergistic interaction was detected.

Animals↗

PubMed2003

Reduction of bacteria on pork carcasses associated with chilling method.

In addition to reducing the temperature of pork carcasses immediately after slaughter and before fabrication, blast chilling (snap chill) or conventional chilling can reduce bacterial populations associated with fresh meats. However, there is little information on bacteria survival resulting from the freeze or chill injury of meat products. In this study, porcine fecal slurries with and without pathogens (Listeria monocytogenes, Salmonella Typhimurium, and Campylobacter coli) were inoculated onto skin-on and skin-off pork surfaces and subjected to industry-specific blast or conventional chilling conditions. A thin agar layer method was used for the recovery of freeze- or chill-injured cells. Test results indicated that there were no statistically significant (P > 0.05) differences between blast and conventional chilling treatments with respect to the reduction of high and low inoculation levels of mesophilic aerobic bacteria, total coliforms, or Escherichia coli on either skin-on or skin-off surfaces. Chilling treatments did not differ significantly (P > 0.05) with respect to their ability to reduce low (3 log10 CFU/cm2) levels of L. monocytogenes and Salmonella Typhimurium. However, C. coli was reduced to undetectable levels, even after enrichment, on pork surfaces inoculated with low levels (3 log10 CFU/cm2) and subjected to blast chilling. Blast and conventional chilling treatments were more effective against all pathogenic bacterial populations when pork surfaces where inoculated at high levels (5 log10 CFU/cm2). The effects of chilling techniques on microbial populations could provide pork processors with an additional intervention for pork slaughter or information to modify and/or improve the chilling process. The information obtained from this study has the potential to serve as a means of producing a microbiologically safer product.

Abattoirs↗

PubMedDate not supplied

In vitro evaluation of the implant-abutment bacterial seal: the locking taper system.

PURPOSE: To test in vitro whether the seal provided by the locking taper used in the implant-abutment connection was capable of preventing the invasion of oral microorganisms. MATERIALS AND METHODS: Twenty-five wide-body implants (5 x 11 mm) and 25 abutments were divided into 2 groups for a 2-phase experiment. The first phase tested the ability of the seal to shield the implant well from outside bacteria; the second phase tested the ability of the seal to prevent bacteria present in the implant well from seeping out. For phase 1, 10 implant-abutment units were immersed in a bacterial broth for 24 hours. The abutments were then separated from the implants and bacterial presence was evaluated using scanning electron microscopy. In phase 2, the tested abutments were inoculated with a droplet of soft agar bacterial gel and assembled with the implant. These units were incubated in a sterile nutrient broth for 72 hours, sampled, and plated to assess bacterial presence. RESULTS: In phase 1, no bacteria were detected in any of the implant wells. In phase 2, no bacteria were detected in the nutrient broth or on the agar plates at 72 hours. DISCUSSION: In implants where a microgap is present, microbial leakage could lead to inflammation and bone loss; thus, it is important to minimize bacterial presence in and around the the implant-abutment junction. CONCLUSION: The seal provided by the locking taper design has been demonstrated to be hermetic with regard to bacterial invasion in vitro.

Colony Count, Microbial↗

PubMed1991

[Microcalorimetric study of the kinetics of the antibacterial effect of third generation cephalosporins in an in vitro dynamic system].

The antimicrobial effect (AME) kinetics of cefotaxime and ceftizoxime was studied with respect to 4 bacterial strains (E. coli, S. aureus, K. pneumoniae and P. aeruginosa) in an in vitro dynamic model. The mean integral concentrations, i.e. the AUC values divided by the dosing interval observed in blood of humans after the drug single intravenous administration in doses of 0.25, 0.5, 1 2 and 4 g. were simulated. The simulated concentrations of cefotaxime and ceftizoxime were 6.25 and 6.75, 12.5 and 13.5, 25 and 27, 50 and 54 and 100 and 108 mg/kg, respectively accounting for the differences in their pharmacokinetics. The changes in the microbial count were recorded microcalorimetrically by the rate of heat output with the BioActivity Monitor LKB 2277-202. The AME was expressed by TE reflecting the shift of the microbial growth curve in the presence of the cephalosporins against the control growth curve (in the absence of the drugs). It was noted that in simulating the concentrations observed after the drugs administration in various doses cefotaxime was mainly superior to ceftizoxime in terms of the TE, the value of the TE correlating with the respective values of the MICs. Some discrepancies between the TE and MICs were explained by the complicated shape of the TE vs. concentration or AUC curve observed earlier in regard to cefotaxime as well as by the differences in the steady state and dynamic conditions of the experiment.

Calorimetry↗

PubMed1996

Microbial densities and physicochemical quality of some crude oil flow stations' saver pit effluents in the Niger Delta areas of Nigeria.

Five crude oil flow stations saver-pit effluents were sampled during the months of October, November, December and January to determine some of their physicochemical qualities and microbiological densities before their discharge into the environment. Standard analytical techniques were employed in the investigation. A high microbial density in the range of 10(6) cfu/ml was observed in the samples. The high coliform count obtained poses a potential threat to the local inhabitants of the area. The physicochemical qualities of the effluents appear to fall within or near acceptable standards. Nevertheless, adequate treatment of the effluent is required especially during the month of January before its discharge into the environment.

Colony Count, Microbial↗

PubMed1994

Use of the strip mutans test in the assessment of caries risk in a group of preschool children.

The purpose of this study was to evaluate the use of a chair-side test involving a count of salivary mutans streptococci (the Strip mutans test) in the assessment of caries risk in a group of preschool children living in an area with a low caries prevalence. A group of 528 4-year-old children were randomly allocated to a study or a control group. In the study group, the baseline microbial data, together with clinical findings of past caries experience, were used for caries risk assessment and for planning subsequent preventive treatment. All children were examined at baseline and after 2 years. Caries experience was assessed according to WHO criteria. There was no difference in caries experience between the study group and the control group at baseline. Within the study group, caries increment was positively correlated (P < 0.01) with the number of mutans streptococci in saliva at baseline, and children assessed 'at risk' at baseline (Strip mutans score > or = 2 and/or > or = 1 dmfs) developed more new lesions than those considered as 'low risk' (mean dmfs 2.6 v 0.9; P < 0.05). The sensitivity and specificity of this combined clinical and microbial caries risk selection were 67% and 75%, respectively, disease being defined as an increment of at least one carious lesion over the 2-year period. In both groups, 50% of the children remained caries inactive during the study. The mean caries increment was, however, lower in the study group than in the control group (mean dmfs 1.7 v 2.1) but the difference was not statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗