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Long-term treatment with subantimicrobial dose doxycycline exerts no antibacterial effect on the subgingival microflora associated with adult periodontitis.

BACKGROUND: The purpose of this study was to determine whether treatment with subantimicrobial dose doxycycline (SDD), 20 mg bid, exerted an antimicrobial effect on the microflora associated with adult periodontitis. METHODS: Following the approval of the protocol and informed consent forms by the respective IRBs at the University of Florida and West Virginia University, 76 subjects with adult periodontitis were entered and randomly assigned to receive SDD or placebo. A split-mouth design was utilized, with each subject receiving subgingival scaling and root planing (SRP) in two quadrants immediately following baseline data collection, while the remaining two quadrants were left unscaled (non-SRP). Microbial samples were collected prior to treatment, after 3, 6, and 9 months of treatment, and after 3 months of no treatment. The samples were examined by microscopy and by enumeration on selective and non-selective media. RESULTS: All treatments resulted in statistically significant decreases in the proportions of spirochetes and motile rods (P <0.05) and in an increase in the proportion of coccoid forms (P <0.0001) relative to baseline. No between-treatment differences were detected between the SDD and placebo treatments in either the SRP or non-SRP design, with the exception of the small and large spirochetal groups. The spirochetal proportions present in the SDD group were significantly lower (P<0.05) than the paired placebo group during the 9-month treatment and was preceded by a significant decrease (P<0.01) in the proportion of microbiologic sample sites that bled on probing. No between- treatment differences were detected in any of the other microbial parameters. CONCLUSION: The microbial differences observed were attributed to the anticollagenase and anti-inflammatory properties of SDD and not to an antimicrobial effect.

Adult↗

Microbiological profiles, pH, and titratable acidity of chorizo and salchichón (two Spanish dry fermented sausages) manufactured with ostrich, deer, or pork meat.

Microbial counts, pH, and titratable acidity were determined in 102 Spanish dry fermented sausages (chorizo and salchichón) made with ostrich, deer, or pork meat. Average microbial counts (log CFU per gram) varied from 5.46 +/- 0.24 to 8.25 +/- 0.80 (total viable counts), from 4.79 +/- 0.36 to 7.99 +/- 0.20 (psychrotrophs), from 0.00 +/- 0.00 to 0.99 +/- 1.10 (undetectable values were assumed to be zero) (Enterobacteriaceae), from 0.00 +/- 0.00 to 4.27 +/- 1.47 (enterococci), from 5.15 +/- 1.15 to 8.46 +/- 0.49 (lactic acid bacteria), from 3.08 +/- 0.44 to 6.59 +/- 1.76 (Micrococcaceae), from 2.27 +/- 1.53 to 5.11 +/- 1.81 (molds and yeasts), from 0.00 +/- 0.00 to 2.25 +/- 0.81 (pseudomonads), and from 0.00 +/- 0.00 to 2.78 +/- 0.46 (Brochothrix thermosphacta). Average pH and titratable acidity varied from 5.07 +/- 0.25 to 5.63 +/- 0.51 (pH units) and from 0.30 +/- 0.01 to 0.86 +/- 0.19 (% lactic acid). Both type of sausage (P < 0.05) and species of meat (P < 0.001) influenced microbial counts. Salchich6n samples showed lower average values than chorizo samples for most microbial groups (significant for Enterobacteriaceae, lactic acid bacteria, and B. thermosphacta) and titratable acidity. Sausages made from pork showed the highest microbial loads for total viable counts, psychrotrophs, Enterobacteriaceae, enterococci, lactic acid bacteria, and yeasts and molds. Higher counts were observed only for pseudomonads in ostrich sausages. B. thermosphacta levels were similar for all species of meat. The highest average pH value was observed in sausages made from ostrich meat, and the lowest titratable acidity level was found in pork sausages.

Animals↗

Effects of elevated atmospheric CO2 concentrations on soil microorganisms.

Effects of elevated CO(2) on soil microorganisms are known to be mediated by various interactions with plants, for which such effects are relatively poorly documented. In this review, we summarize and synthesize results from studies assessing impacts of elevated CO(2) on soil ecosystems, focusing primarily on plants and a variety the of microbial processes. The processes considered include changes in microbial biomass of C and N, microbial number, respiration rates, organic matter decomposition, soil enzyme activities, microbial community composition, and functional groups of bacteria mediating trace gas emission such as methane and nitrous oxide. Elevated CO(2) in atmosphere may enhance certain microbial processes such as CH(4) emission from wetlands due to enhanced carbon supply from plants. However, responses of extracellular enzyme activities and microbial community structure are still controversy, because interferences with other factors such as the types of plants, nutrient availabilitial in soil, soil types, analysis methods, and types of CO(2) fumigation systems are not fully understood.

Atmosphere↗

Characterization of naturally developing small intestinal bacterial overgrowth in 16 German shepherd dogs.

Sixteen German Shepherd Dogs were found, via quantitative microbial culture of intestinal fluid samples, to have small intestinal bacterial overgrowth (IBO) over an 11-month period. All dogs were deficient in serum IgA. Consistent clinical signs suggestive of an alimentary tract disorder were not observed. Serum cobalamin determinations were not helpful in detecting IBO. Serum folate concentrations had variable sensitivity and specificity for detecting dogs from which we could culture > or = 1 x 10(5) bacterial/ml from intestinal fluid samples in the nonfed state. Histologic and intestinal mucosal cytologic examinations were not useful in detecting IBO. Substantial within-dog and between-dog variation was found in the numbers and species of bacteria in the intestines. The difficulty in diagnosing IBO, the variability in organisms found in individual dogs on repeated sampling, the likelihood that intestinal fluid microbial cultures failed to diagnose IBO in some dogs, and the potential of IBO to be clinically inapparent were the most important findings in this study.

Animals↗

Microbial barrier properties of hen egg shells.

Scanning electron micrographs of shell surfaces revealed a highly fissured outer layer with few open pores on brown eggs but many on white eggs. Total removal of the cuticle with solvents was difficult but partial removal with surface etching was possible using concentrated nitric acid. Staining methods to estimate the number of pores were unsatisfactory but it was possible to detect and count pores on micrographs, as pore mouths were usually associated with depressions and cuticle disruptions. Additionally, porosity could be estimated by measuring the distance between pores along fractured edges of samples. Egg shell contents were replaced with nutrient agar and incubated in Escherichia coli broth. Colonies were subsequently isolated from the agar, indicating that the barrier properties of shells and membranes had been compromised. Experiments with isolated inner shell membranes showed that these posed no significant barrier to E. coli. The exposure of whole intact eggs to E. coli broth followed by seep filtration and microbiological analysis of egg contents, indicated that bacteria had entered the eggs. The degree of infection was correlated with the pole of the egg in contact with the E. coli broth and was attributed to the increased porosity of the blunt pole of the egg compared with that of the apex. Similar experiments immersing eggs into broth inoculated with a Salmonella strain resulted in contamination of the egg contents with this organism.

Animals↗

Microbiological status of commercially available medicinal herbal drugs--a screening study.

One hundred and thirty-eight medicinal herbal drugs obtained from different suppliers were examined for microbial contaminants and for the detectability of pathogenic microorganisms. For this purpose, several microbiological standard parameters (total aerobic mesophilic count, enterobacteria, coliforms, aerobic sporeformers, yeasts and moulds, enterococci, lactobacilli, pseudomonades and aeromades) and selective methods for the detection of indicator microorganisms and pathogens (E. coli, enterohaemorrhagic E. coli [EHEC], Salmonella, Campylobacter jejuni, Pseudomonas aeruginosa, Bacillus cereus, Clostridium perfringens, Listeria, coagulase-positive staphylococci, Candida albicans, potentially aflatoxigenic moulds) were applied. The microbial load of the samples varied considerably. While none of the samples contained EHEC, Salmonellae, Pseudomonas aeruginosa, Listeriae, Staphylococcus aureus or Candida albicans, four samples were E. coli positive, two samples were presumptively Campylobacter jejuni positive and nine herbal drugs contained a potentially aflatoxigenic mould flora. Further details regarding different viable count classes as well as preparation techniques are discussed.

Bacteria↗

Clinical significance of quantitative blood cultures in newborn infants.

AIM: To evaluate quantitative blood culture as a secondary test on a positive blood culture for the diagnosis of sepsis in newborn infants. METHOD: A 15-month prospective study of colony forming units (CFU) on positive blood cultures from newborn infants clinically suspected of having bacterial sepsis. Growth of bacteria in peripheral blood cultures was quantified using the isolater 1.5 microbial tube lysis direct plating culture system. Colony forming units were evaluated against a clinical assessment of infection. RESULTS: Of 137 positive blood cultures, 71 (51.8%) were taken from neonates with clinically defined infection and 66 (48.2%) were from non-infected infants. The clinical and biographical data in these two groups were similar. Coagulase negative staphylococci were the most commonly isolated organisms in each group (60.6% vs 93.9%). Eight deaths from sepsis occurred in the clinically infected group. Eighty-five per cent of sepsis was late onset. Although a CFU count > or = 30/mL predicted sepsis (sensitivity 83%, specificity 60%, positive predictive value 69%, negative predictive value 76%), a CFU count < 30/mL did not rule out serious sepsis. The higher the CFU count the greater the likelihood of sepsis. CONCLUSION: Quantitative blood culture was not shown to be a sensitive secondary test on a positive blood culture to distinguish clinical sepsis from culture contamination. Although a positive threshold of > or = 30 CFU/mL proved to be optimal, improvement in test performance would be expected with a lower incidence of culture contamination.

Bacteremia↗

Effects of periodontitis and smoking on the microbiota of oral mucous membranes and saliva in systemically healthy subjects.

BACKGROUND/AIM: To examine the differences in the proportions of 40 bacterial species in samples from eight oral soft-tissue surfaces and saliva in systemically healthy adult subjects with and without periodontitis and in smokers and nonsmokers. METHODS: Saliva and microbial samples were taken from eight oral soft-tissue surfaces in 229 systemically healthy subjects stratified by periodontal and smoking status. Subjects included 84 periodontally healthy and 145 periodontitis individuals. Of these, there were 182 nonsmokers and 47 smokers. Soft-tissue samples were taken using a "buccal brush" and saliva was taken by expectoration. All samples were individually evaluated for their content of 40 bacterial species using checkerboard DNA-DNA hybridization. The percentage of total DNA probe count was determined for each species at each sample location and averaged across subjects. The significance of differences among the proportions of the 40 test species at different sample locations was sought in these populations using the Kruskall-Wallis test and adjusted for multiple comparisons. RESULTS: When stratified for periodontal status or smoking status, microbial profiles at most sites did not differ significantly. Generally, periodontal pathogens were found at higher levels on the soft tissues of periodontitis subjects than in periodontally healthy subjects, and at higher levels in smokers than nonsmokers. Few significant differences were found, although trends in the populations were noted. CONCLUSIONS: Small differences in the proportions of bacterial species were observed on soft tissues and in saliva comparing periodontally healthy and periodontitis subjects. Somewhat greater differences between smokers and nonsmokers were found; however, these were not statistically significant.

Actinomyces↗

[The microbial count of insulin multiple-dose containers].

In accordance with the Pharmacopoeia of the GDR, from multiple-dose containers containing injection preparations with preservatives added such preparations may be repeatedly withdrawn up to 72 h after opening these containers. Exempted from this limitation are multiple-dose containers containing insulin injection preparations repeatedly used by diabetics for self-injections. Microbiological tests were to contribute to clearing the question whether the period of using these insulin multiple-dose containers can be provided to be maximally 30 d after first use.

Colony Count, Microbial↗

Fungal contribution to in situ biodegradation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) film in soil.

The contribution of fungi to the microbial degradation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) films in soil was studied. Various traces, cavities, and grooves observed on the dented surface of PHBV films demonstrated that the degradation was a concerted effect of a microbial consortium colonizing the film surface, including fungi, bacteria, and actinomycetes. The succession of microbial consortia in the soil around the PHBV films during the degradation showed a distinctive increase in the fungal population, resulting in its dominance. Comparison of the degradation ability of microbial strains isolated from soil where PHBV films were degraded, revealed that fungi showed the highest contribution to PHBV degradation, growing very rapidly along the film surface with their high degradation ability and then expanding their hyphae in a three-dimensional manner.

Bacteria↗

Screening for microbial markers in Miocene sediment exposed during open-cast brown coal mining.

Viable microorganisms were found in Miocene lacustrine clays of the cypris formation excavated from 200-m below the surface as spoil during open-cast brown coal mining (Sokolov Brown Coal Basin, North-Western Bohemia, Czech Republic). Both saprotrophic microfungi of the genera Penicillium, Verticillium, Cladosporium and Aspergillus as well as heterotrophic bacteria were isolated from an intact sediment cores. Heterotrophic bacteria were classified by the MIS Sherlock System as representatives of genera Nocardiopsis, Arthrobacter, Micrococcus, Kocuria, Rothia, Clavibacter, Bacillus, Paenibacillus, Brevibacillus, Microbacterium, Acinetobacter and Pseudomonas. A bacterium found among the strains had an atypical fatty acids profile enriched by branched fatty acids and polyunsaturated fatty acid (18:3 omega 6) and gave no MIS Sherlock match. Phospholipid fatty acids analysis indicates a relatively high (535 pmol g(-1)) but inhomogeneously distributed viable microbial biomass. Fatty acids analyses of non-fractioned lipids (representing viable, storage and dead biomass; 8390 pmol g(-1)) detected rich and homogenous profiles with fungal, bacterial and actinomycetal markers but no protozoan and algal fatty acids markers.

Bacteria↗

Identification of dominant bacteria in feces and colonic mucosa from healthy Spanish adults by culturing and by 16S rDNA sequence analysis.

The aim of this work was to examine by culturing the changes in the total and indicator populations of the feces of two individuals over 1 year and to identify the dominant microbial components of a single sample of feces from each donor. Populations and dominant bacteria from a sample of colonic mucosa from a further individual were also assessed. The culture results were then compared to those obtained with the same samples by 16S rDNA cloning and sequencing. High interindividual variation in representative microbial populations of the gastrointestinal tract (GIT) was revealed by both the culture and the culture-independent techniques. Species belonging to Clostridium clusters (XIVa, IV, and XVIII) predominated in both the fecal and the mucosal samples (except in the mucose cultured isolates), members of Clostridium coccoides cluster XIVa being the most numerous microorganisms. Species of gamma-proteobacteria (Escherichia coli and Shigella spp.), bifidobacteria, and actinobacteria appeared in lower numbers than those of clostridia. From the mucosal cultured sample, only facultative anaerobes and bifidobacteria were recovered, suggesting destruction of the anaerobe population during processing. In accordance with this, the microbial diversity revealed by 16S rDNA sequence analysis was greater than that revealed by culturing. Despite large interindividual differences, distinct human communities may have group-associated GIT microbiota characteristics, such as the low number of Bacteroides seen in the subjects in this study.

Adult↗

Effect of microbial contamination and pH changes in storage solutions during in vitro assays of bonding agents.

A study of pH changes in, and microbial colonization of, storage solutions being utilized during an in vitro evaluation of luting agent solubility has been made. Microbial growth in storage solutions was observed to occur at as high as 10(5) colony-forming units (cfu)/mL after seven days' incubation. A strong time-dependent inverse correlation between pH and cfu was found, with the pH dropping to as low as 5.3 (p less than 0.01). Decreases in pH were not found in presterilized control samples (p less than 0.05). Further, it was demonstrated that presterilization with ethylene oxide or the addition of respiratory inhibitors prevented both microbial colonization and the pH drop when compared with controls (p less than 0.001). These data suggest that studies of dental materials which include incubations in various fluids should be controlled for the potential effects of micro-organisms and their effect on pH.

Colony Count, Microbial↗

An epidemiological study assessing the relative importance of airborne and direct contact transmission of microorganisms in a medical intensive care unit.

A prospective epidemiological survey was carried out over a period of seven weeks in a medical intensive care unit. Bacteria from patients, staff and air were monitored and the transmission of isolated microorganisms was followed. Handwashing samples revealed pathogenic bacteria in 30.8% of physicians (average number of colony forming units: 71,300 per hand) and 16.6% of nurses (39,800 cfu per hand). Air cultures yielded pathogens in 15% of sampling periods and nine of 53 patients were found to be colonized with Gram-negative bacteria, Staphylococcus aureus or Candida spp. The spectrum of bacteria recovered from patients and air was generally different, whereas strains recovered from patients and their attendants' hands were indistinguishable on multiple occasions. The results of this study confirm that direct contact is the principal pathway of microbial transmission, whereas little evidence for a significant role of airborne transmission is shown. The call for more extensive air-filtering and ventilation systems in medical intensive care units is not supported by the results shown in this communication.

Air Microbiology↗

Colonization and epithelial adhesion in the pathogenesis of neonatal candidiasis.

Candida species are important nosocomial pathogens in the newborn population, particularly among the premature very-low-birth-weight infants in neonatal intensive care units. Candida colonization of the neonatal skin and gastrointestinal tract is an important first step in the pathogenesis of invasive disease. C albicans is the most commonly isolated species in colonized or infected infants. Over the past decade the incidence of both colonization and infection with other Candida species, particularly C parapsilosis, has risen dramatically. Colonization of the infant occurs early in life and is affected by a variety of common practices in neonatal intensive care. Microbial factors also augment colonization, including the ability of Candida to adhere to human epithelium. A better understanding of the complex interactions between host risk factors and virulence traits of colonizing yeast may allow the risk of systemic spread to be reduced in the population of premature infants.

Bacterial Adhesion↗

The effect of a toothpaste containing 2% zinc citrate and 0.3% Triclosan on bacterial viability and plaque growth in vivo compared to a toothpaste containing 0.3% Triclosan and 2% copolymer.

OBJECTIVE: To compare the antimicrobial efficacy and effect on plaque growth of a new silica-based fluoride toothpaste containing 2% zinc citrate/ 0.3% Triclosan with a silica-based fluoride toothpaste containing 0.3% Triclosan/2% copolymer. METHODS: In Study 1, plaque was collected after one week's use of each toothpaste and assessed for bacterial viability, live/ dead ratio and microbial membrane integrity. In study 2, plaque was measured immediately and 18 hours after a single brushing with the specified toothpastes. RESULTS: The 2% zinc citrate/0.3% Triclosan formulation significantly reduced the total number of viable aerobic and anaerobic bacteria (p = 0.0223 and p = 0.0443 respectively) compared to the 0.3% Triclosan/2% copolymer formulation. Both toothpastes increased the bacterial membrane permeability significantly. However, the proportion of live bacteria for the 2% zinc citrate/0.3% Triclosan product was significantly reduced (p < 0.05). Study 2 showed significantly less plaque growth 18 hours after using the 2% zinc citrate/0.3% Triclosan toothpaste compared to the 0.3% Triclosan/2% copolymer toothpaste (p < 0.01). CONCLUSION: Regular use of a fluoride toothpaste containing 2% zinc citrate and 0.3% Triclosan, significantly reduced the viability of plaque bacteria compared to a fluoride toothpaste containing 0.3% Triclosan/ 2% copolymer 12 hours after brushing. In addition, a clinical plaque growth study confirmed that this anti-microbial efficacy leads to a significant reduction in plaque growth.

Adult↗

Microbial community composition affects soil fungistasis.

Most soils inhibit fungal germination and growth to a certain extent, a phenomenon known as soil fungistasis. Previous observations have implicated microorganisms as the causal agents of fungistasis, with their action mediated either by available carbon limitation (nutrient deprivation hypothesis) or production of antifungal compounds (antibiosis hypothesis). To obtain evidence for either of these hypotheses, we measured soil respiration and microbial numbers (as indicators of nutrient stress) and bacterial community composition (as an indicator of potential differences in the composition of antifungal components) during the development of fungistasis. This was done for two fungistatic dune soils in which fungistasis was initially fully or partly relieved by partial sterilization treatment or nutrient addition. Fungistasis development was measured as restriction of the ability of the fungi Chaetomium globosum, Fusarium culmorum, Fusarium oxysporum, and Trichoderma harzianum to colonize soils. Fungistasis did not always reappear after soil treatments despite intense competition for carbon, suggesting that microbial community composition is important in the development of fungistasis. Both microbial community analysis and in vitro antagonism tests indicated that the presence of pseudomonads might be essential for the development of fungistasis. Overall, the results lend support to the antibiosis hypothesis.

Antibiosis↗

[Micro-ecological aspects of studies on the effect of antibacterial drugs on intestinal microflora].

Possible characterization of intestinal microflora as an integral system after exposure to antibacterial drugs was studied. Microflora of the contents and mucosa of the jejunum and large intestine in control rats and in rats exposed to metronidazole was studied and numerical indicators characterizing ratios of dominating and accompanying microbial groups in the intestine biotope++ were developed. With this purpose the proportion of the microbial groups in the total quantity of the microbes of a microbiocenosis was determined by the data on microflora quantitative composition. On the basis of detected wide species variety of microorganisms potentially dominating by their biotope numerical limits of the norm were determined only for the microbial groups of the accompanying microflora. The total proportion of the accompanying microbial populations under the normal conditions and the detected measure of deviation (reverse, partial) from the ratio of the dominating and accompanying microorganisms peculiar of the given biotope++ in separate subjects promoted estimation of microbiocenoses of definite biotope of the intestine as a whole.

Animals↗