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The microbiological status of patients with periodontitis in southern Estonia after non-surgical periodontal therapy.

Our objective was to investigate the presence of periodontal pathogens in comparison with the total degree of microorganisms after non-surgical periodontal therapy. The study material consisted of microbiological samples from periodontal pockets originating from 140 consecutive patients with chronic generalized severe periodontitis. The subgingival samples from periodontal pockets were obtained by a sterile curette, placed into 2 ml of the VMGA III medium, homogenized and serially diluted in the Brucella broth. 100 microl aliquots from the dilutions were inoculated onto the Brucella and the TSBV agar. The plates were incubated in an anaerobic chamber and under microaerobic conditions. The isolates were identified according to colonial and cellular morphology, the potency disk pattern, and the biochemical profiles. After instrumentation, no periodontal pathogens were isolated in 46 (33%) patients, while 94 patients (67%) were infected with one to five different periodontal pathogens. However, higher degree of the total microflora was positively correlated with number of isolated pathogens, a putative indicator of their presence. Therefore, due to the occurrence of residual microorganisms after non-surgical mechanical treatment, information about the pattern of these pathogens is needed for application of antimicrobial therapy. The level of microbial load in gingival pockets, including both pathogenic and non-pathogenic species, is one of the determinants of presence of residual pathogens after non-surgical periodontal therapy.

Bacteria, Aerobic↗

Bacterial contamination of hospital pagers.

We assessed the bacterial contamination of the pagers of healthcare personnel and the efficacy of disinfection with 70% isopropyl alcohol. Microorganisms were isolated from all pagers; 21% yielded Staphylococcus aureus, of which 14% were methicillin resistant. Cleaning with alcohol reduced the total colony count by an average of 94%. Bacterial load varied by healthcare worker group and service assignment.

2-Propanol↗

Increased ethyl mercury load in protein a immunoadsorption.

Immunoadsorption is an adsorption technique for extracorporeal removal of circulating autoantibodies in autoimmune diseases. To prevent microbial growth during storage, the protein A columns are primed with thiomersal, which contains toxic ethyl mercury, which may be released during the procedure and potentially begin to accumulate and become toxic. To reduce the thiomersal-related mercury release during immunoadsorption treatment, we introduced a modified rinsing solution containing N-acetylcysteine, which is an avid mercury scavenger. Thirteen patients received 17 protein A immunoadsorption treatments and their venous blood samples were collected immediately before and after each session. The total blood mercury levels were measured by atomic absorption spectrometry, and the ethyl mercury levels by atomic fluorescence spectrometry. Following the manufacturer's recommendations, we used 600 mg of N-acetylcysteine to rinse the mercury from protein-loaded columns before each immunoadsorption treatment. After immunoadsorption, the ethyl mercury levels increased from 0.148 +/- 0.402 ng/g to 2.026 +/- 1.944 ng/g (P < 0.001), and the total blood mercury levels increased from 2.447 +/- 3.065 ng/g to 20.437 +/- 28.603 ng/g (P = 0.02). The post-treatment values of total blood mercury exceeded the upper safety level of 5 ng/g in all 17 immunoadsorption treatments, but no patient developed clinical signs of mercury toxicity. The results of our study showed an increase in total blood mercury and ethyl mercury levels during the immunoadsorption treatments, suggesting mercury release from thiomersal-primed columns despite the addition of N-acetylcysteine to the rinsing solution.

Acetylcysteine↗

Production of PHA from starchy wastewater via organic acids.

Polyhydroxyalkanoate (PHA) was produced from a starchy wastewater in a two-step process of microbial acidogenesis and acid polymerization. The starchy organic waste was first digested in a thermophilic upflow anaerobic sludge blanket (UASB) reactor to form acetic (60-80%), propionic (10-30%) and butyric (5-40%) acids. The total volatile fatty acids reached 4000 mg l(-1) at a chemical oxygen demand (COD) loading rate of 25-35 g l(-1) day(-1). A carbon balance indicates that up to 43% of the organic carbon in the starchy waste went to the organic acids and the rest to biogas, volatile suspended solids and residual sludge accumulated in the reactor. The acid composition profile was affected by COD loading rate: a medium rate around 9 g l(-1) day(-1) gave a high propionic acid content (29% wt) and a high rate around 26 g l(-1) day(-1) led to a high butyric acid content (34% wt). The acids in the effluent solution after microfiltration were utilized and polymerized into PHA by bacterium Alcaligenes eutrophus in a second reactor. Fifty grams of PHA was produced from 100 g total organic carbon (TOC) utilized, a yield of 28% based on TOC, which is comparable with 55 g PHA per 100 g TOC of pure butyric and propionic acids used. PHA formation from individual acids was further investigated in a semi-batch reactor with three acid feeding rates. With a limited nitrogen source (80-100 mg NH(3) per liter), the active biomass of A. eutrophus, not including the accumulated PHA in cells, was maintained at a constant level (8-9 g l(-1)) while PHA content in the cell mass increased continuously in 45 h; 48% PHA with butyric acid and 53% PHA with propionic acid, respectively. Polyhydroxybutyrate was formed from butyric acid and poly(hydroxybutyrate-hydroxyvalerate) formed from propionic acid with 38% hydroxyvalerate.

Alcaligenes↗

Application of a solvent-tolerant microbial consortium for biofiltration of extremely high concentration gaseous solvent streams.

The aerobic biological oxidation of 2-propanol (isopropyl alcohol, IPA) at extremely high concentrations in air by an enriched solvent-tolerant microbial consortium operating at ambient temperature was evaluated for six months. Solvent-tolerant microbial cells were immobilised onto porous glass pall rings and fed with either IPA or its metabolic product acetone as sole carbon source. Successful biofiltration of solvent vapour at a concentration of 24 g m(-3) was achieved with oxidation of up to 100% total inlet carbon. The maximum IPA mass loading and IPA elimination capacity (EC) was 1700 g m(-3) h(-1). This performance exceeds all previous values published in the literature for similar processes. A slip feed experiment, using acetone, was also performed in order to assess the substrate specificity performance. The biofilter responded successfully to a switch from acetone to IPA as sole carbon source, displaying little reduction in overall organic carbon removal.

2-Propanol↗

[Leukocyte agglutination test for the determination of microbial burden].

Transfer of laboratory animals from the breeder to the experimental unit includes in most cases a change of the microbial environment even under SPF-conditions. Many experimental treatments may also disturb the balance between host and microbial load and provoke infections. With the aim to detect such undesired effects in the course of the experiment the suitability of the leukergy test was investigated. SPF-rats, endotoxin-treated rats, and conventionalized germ-free rats were checked for leukocyte agglomeration, and the findings were compared with total leukocyte number, blood picture, endotoxin content in the blood and body temperature. A rise of leukergy from 10 to more than 20% was recorded in endotoxin-treated and conventionalized rats.

Agglutination↗

[The use of neck skin for microbial process control of fresh poultry meat using the bioluminescence method].

In this study comparative investigations of the total viable counts of bacteria and viable counts of Enterobacteriaceae with the ATP content were carried out on 70 poultry carcasses and neck skins thereof. The results revealed a correlation of total viable counts on the neck skin and on the carcass sample. Therefore the examination on total viable counts of neck skin is able to give results on the respective load of the carcass (coefficient of correlation (r) = 0.85 and coefficient of determination (R2) = 0.71). This only applies, however, to a slaughter technology at which the poultry hangs up site down with the neck skin to the bottom. The results of the ATP investigation expressed in Relative Light Units (RLU) in a bioluminescence method and total viable counts of neck skin samples are correlated (r = 0.85; R2 = 0.72). Lower values for r and R2 were estimated for RLU/Enterobacteriaceae on neck skin (r = 0.64; R2 = 0.41), for RLU/total viable counts on the carcass (r = 0.66; R2 = 0.46) and for RLU/Enterobacteriaceae on the carcass (r = 0.33; R2 = 0.11). To estimate the hygiene status in poultry slaughtery, sampling from the neck skin can replace the sampling on the whole carcass The bioluminescence method is suitable to replace the determination of total viable counts in the context of in-house hygiene supervision. However, this method seemed to be less reliable to predict the Enterobacteriaceae counts.

Abattoirs↗

Performance of subsurface flow constructed wetland taking pretreated swine effluent under heavy loads.

Subsurface flow constructed wetlands (SSFCW) subjected to changing of loading rates are poorly understood, especially when used to treat swine waste under heavy loads. This study employed a SSFCW system to take pretreated swine effluent at three hydraulic retention times (HRT): 8.5-day HRT (Phase I), 4.3-day HRT (Phase II), and 14.7-day HRT (Phase III). Results showed that the system responded well to the changing hydraulic loads in removing suspended solids (SS) and carbonaceous oxygen demands. The averaged reduction efficiencies for four major constituents in the three phases were: SS 96-99%, chemical oxygen demand (COD) 77-84%, total phosphorus 47-59%, and total nitrogen (TN) 10-24%. While physical mechanisms were dominant in removing pollutants, the contributions of microbial mechanisms increased with the duration of wetland use, achieving 48% of COD removed and 16% of TN removed in the last phase. Water hyacinth made only a minimal contribution to the removal of nutrients. This study suggested that the effluent from SSFCW was appropriate for further treatment in land applications for nutrient assimilation.

Animals↗

Fermentation of yam: microbiology and sensory evaluation of cooked fermented yam tissues.

The microflora of white yam (Dioscorea rotundata L.) slices fermented anaerobically in 1.5 percent brine for five days at room temperature were studied. The hydrolysis of the carbohydrate and the subsequent conversion of sugars and minerals by the fermenting microbes contributed much to the increased microbial load especially within the first 72 hours of fermentation. The organisms implicated in the fermentation include the species of Pediococcus, Lactobacillus and Pseudomonas, Bacillus subtilis and two other Gram negative coccal cells, yet to be identified. The decrease in microbial counts at the latter stage of fermentation was attributed to the high total acidity of the medium, which was about 2.67 times the initial value of 0.027% lactic acid. Meanwhile, the lactic acid bacteria increased continuously throughout the period of fermentation.

Cooking↗

Aerobic spore-forming bacteria and chemical composition of some Nigerian fermented soup condiments.

A total of 97 strains of spore-forming Bacillus were isolated from 45 samples of three Nigerian fermented condiments, obtained from retail markets located in Southwestern Nigeria. The isolates were identified as B. subtilis (33%), B. pumilus (19%), B. licheniformis (22%), B. brevis (9%), B. megaterium (12%) and B. polymyxa (5%). The microbial load of the condiments showed that the average count of spore-formers was between 107 to 109 cfu/g. The moisture contents of iru, ugba and ogiri were 57.18%, 46.32% and 42.34%, respectively, while the protein contents were 18.26%, 17.17% and 17.96%. The percentage fat was 29.88%, 40.25% and 44.14% for iru, ugba and ogiri. The ash content ranged from 5.8 to 6.1%; a 0.1% titratable acidity and pH values above 7.0 were obtained for the three condiments.

Bacillus↗

Determination of bacterial load by real-time PCR using a broad-range (universal) probe and primers set.

The design and evaluation of a set of universal primers and probe for the amplification of 16S rDNA from the Domain Bacteria to estimate total bacterial load by real-time PCR is reported. Broad specificity of the universal detection system was confirmed by testing DNA isolated from 34 bacterial species encompassing most of the groups of bacteria outlined in Bergey's Manual of Determinative Bacteriology. However, the nature of the chromosomal DNA used as a standard was critical. A DNA standard representing those bacteria most likely to predominate in a given habitat was important for a more accurate determination of total bacterial load due to variations in 16S rDNA copy number and the effect of generation time of the bacteria on this number, since rapid growth could result in multiple replication forks and hence, in effect, more than one copy of portions of the chromosome. The validity of applying these caveats to estimating bacterial load was confirmed by enumerating the number of bacteria in an artificial sample mixed in vitro and in clinical carious dentine samples. Taking these parameters into account, the number of anaerobic bacteria estimated by the universal probe and primers set in carious dentine was 40-fold greater than the total bacterial load detected by culture methods, demonstrating the utility of real-time PCR in the analysis of this environment.

Bacteria, Aerobic↗

Multiple microbial activities for volatile organic compounds reduction by biofiltration.

In the northeast of Italy, high volatile organic carbon (VOC) emissions originate from small-medium companies producing furniture. In these conditions it is difficult to propose a single, efficient, and economic system to reduce pollution. Among the various choices, the biofiltration method could be a good solution, because microbial populations possess multiple VOC degradation potentials used to oxidize these compounds to CO2. Starting from the air emissions of a typical industrial wood-painting plant, a series of experiments studied in vitro microbial degradation of each individual VOC. Isolated strains were then added to a laboratory-scale biofiltration apparatus filled with an organic matrix, and the different VOC behavior demonstrated the potential of single and/or synergic microbial removal actions. When a single substrate was fed, the removal efficiency of a Pseudomonas aeruginosa inoculated reactor was 1.1, 1.17, and 0.33 g m(-3) hr(-1), respectively, for xylene, toluene, and ethoxy propyl acetate. A VOC mixture composed of butyl acetate, ethyl acetate, diacetin alcohol, ethoxy propanol acetate, methyl ethyl ketone, methyl isobutyl ketone, toluene, and xylene was then fed into a 2-m(3) reactor treating 100 m3 hr(-1) of contaminated air. The reactor was filled with the same mixture of organic matrix, enriched with all of the isolated strains together. During reactor study, different VOC loading rates were used, and the behavior was evaluated continuously. After a short acclimation period, the removal efficiency was > 65% at VOC load of 150-200 g m(-3) hr(-1). Quantification of removal efficiencies and VOC speciation confirmed the relationship among removal efficiencies, compound biodegradability, and the dynamic transport of each mixture component within the organic matrix. Samples of the fixed bed were withdrawn at different intervals and the heterogeneous microbial community evaluated for both total and differential compound counts.

Air Pollutants↗

Retention and removal of the fish pathogenic bacterium Yersinia ruckeri in biological sand filters.

AIMS: To investigate the retention and removal of the fish pathogenic bacterium Yersinia ruckeri in biological sand filters and effects on the microbial community composition. METHODS AND RESULTS: Sand filter columns were loaded (70 mm day(-1)) with fish farm wastewater and a suspension (10(8) CFU ml(-1)) of Y. ruckeri. Bacterial numbers and protozoan numbers were determined by plate counts and epifluorescence microscopy, respectively, and microbial biomass and community composition were assessed by phospholipid fatty acids (PLFA) analysis. Concentrations of Y. ruckeri in the filter effluent decreased from 10(8) to 10(3)-10(5) CFU ml(-1) during the experiment. Numbers of Y. ruckeri in the sand decreased from 10(6) CFU g(-1) dry weight (DW) sand to 10(4) CFU g(-1) DW sand. In contrast, microbial biomass determined with plate counts and total PLFA increased during the whole experiment. Principal component analysis (PCA) revealed a change in microbial community composition with time, with the most pronounced change in surface layers and towards the end of the experiment. Protozoan numbers increased from ca 0-600 cells g(-1) DW sand, indicating the establishment of a moderate population of bacterial grazers. CONCLUSIONS: The removal of Y. ruckeri improved during the experiment. Introduction of Y. ruckeri to the sand filter columns stimulated growth of other micro-organisms, which in turn caused a shift in the microbial community composition in the sand. SIGNIFICANCE AND IMPACT OF THE STUDY: This study increases the understanding of the dynamics of sand filters subjected to a high loading of a pathogenic bacterium and can therefore be used in future work were the overall aim is to provide a more reliable and efficient removal of pathogenic bacteria in biological sand filter systems.

Animals↗

Influence of biomass production and detachment forces on biofilm structures in a biofilm airlift suspension reactor

The influence of process conditions (substrate loading rate and detachment force) on the structure of biofilms grown on basalt particles in a Biofilm Airlift Suspension (BAS) reactor was studied. The structure of the biofilms (density, surface shape, and thickness) and microbial characteristics (biomass yield) were investigated at substrate loading rates of 5, 10, 15, and 20 kg COD/m3. day with basalt concentrations of 60 g/L, 150 g/L, and 250 g/L. The basalt concentration determines the number of biofilm particles in steady state, which is the main determining factor for the biofilm detachment in these systems. In total, 12 experimental runs were performed. A high biofilm density (up to 67 g/L) and a high biomass concentration was observed at high detachment forces. The higher biomass content is associated with a lower biomass substrate loading rate and therefore with a lower biomass yield (from 0.4 down to 0.12 gbiomass/gacetate). Contrary to general beliefs, the observed biomass detachment decreased with increasing detachment force. In addition, smoother (fewer protuberances), denser and thinner compact biofilms were obtained when the biomass surface production rate decreased and/or the detachment force increased. These observations confirmed a hypothesis, postulated earlier by Van Loosdrecht et al. (1995b), that the balance between biofilm substrate surface loading (proportional to biomass surface production rate, when biomass yield is constant) and detachment force determines the biofilm structure. When detachment forces are relatively high only a patchy biofilm will develop, whereas at low detachment forces, the biofilm becomes highly heterogeneous with many pores and protuberances. With the right balance, smooth, dense and stable biofilms can be obtained. Copyright 1998 John Wiley & Sons, Inc.

Journal Article↗

Thiobacillus sp. W5, the dominant autotroph oxidizing sulfide to sulfur in a reactor for aerobic treatment of sulfidic wastes.

The floating filter technique was successfully adapted for the isolation of the dominant, chemolithoautotrophic, sulfide-oxidizing bacterium from a sulfur-producing reactor after conventional isolation techniques had failed. The inoculated polycarbonate filters, floating on mineral medium, were incubated under gaseous hydrogen sulfide at non-toxic levels. This technique gave 200-fold higher recoveries than conventional isolation techniques. Viable counts on the filters, making up 15% of the total count, appeared to be all of the same species. Chemostat cultures of the new isolate had a very high sulfur-forming capacity, converting almost all hydrogen sulfide in the medium to elemental sulfur under high sulfide loads (27.5 mmol l-1 h-1) and fully aerobic conditions. This behaviour closely resembled that of the microbial community in the sulfur-producing reactor. Moreover, similar protein patterns were obtained by electrophoresis of cell-free extracts from the isolate and the mixed culture. It has therefore been concluded that this isolate represents the dominant sulfide-oxidizing population in the reactor. The isolate has been shown to be a new Thiobacillus species, related to Thiobacillus neapolitanus. In view of the general confusion currently surrounding the taxonomy of the thiobacilli, a new species has not been formally created. Instead, the isolate has been given the working name Thiobacillus sp. W5.

Aerobiosis↗

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↗

Microbial population dynamics during fed-batch operation of commercially available garbage composters.

Microbial populations in terms of quantity, quality, and activity were monitored during 2 months of start-up operation of commercially available composters for fed-batch treatment of household biowaste. All the reactors, operated at a waste-loading rate of 0.7 kg day(-1) (wet wt), showed a mass reduction efficiency of 88-93%. The core temperature in the reactors fluctuated between 31 degrees C and 58 degrees C due to self-heating. The pH declined during the early stage of operation and steadied at pH 7.4-9.3 during the fully acclimated stage. The moisture content was 48-63% early in the process and 30-40% at the steady state. Both direct total counts and plate counts of bacteria increased via two phases (designated phases I, II) and reached an order of magnitude of 10(11) cells g(-1) (dry wt) at the steady state. Microbial community changes during the start-up period were studied by culture-independent quinone profiling and denatured gradient gel electrophoresis (DGGE) of PCR-amplified 16S rDNA. In all the reactors, ubiquinones predominated during phase I, whereas partially saturated menaquinones became predominant during phase II. This suggested that there was a drastic population shift from ubiquinone-containing Proteobacteria to Actinobacteria during the start-up period. The DGGE analysis of the bacterial community in one of the reactors also demonstrated a drastic population shift during phase I and the predominance of members of the phyla Proteobacteria and Bacteroidetes during the overall period. But this molecular analysis failed to detect actinobacterial clones from the reactor at any stage.

Actinobacteria↗

Biological pre-treatment of wastewater containing sulfate using anaerobic immobilized cells.

Biological reduction of sulfate to sulfide using sulfate reducing bacteria (SRB) was investigated. A respirometer was used to study the sulfide toxicity in the systems fed glucose, the results showed that sulfide would start to inhibit methanogens when the dissolved sulfide and total sulfide concentrations were 276.4 and 304.6 mg/L, respectively. When chemostats were used to study the Monod kinetic coefficients, Y, kd, Ks, and k were 0.36 mg VSS (volatile suspended solids) using SRB/mg SO4-S, 0.05/day, 147.30 mg SO4-S/L, and 6.50 mg SO4-S/mg VSS using SRB-d, respectively. Using pure cultural techniques, SRB were found to be 29.45% of the VSS in the chemostats. Sulfate removal using an upflow anaerobic filter packed with immobilized cells was also investigated. Under sulfate loading rates of 0.2 and 0.4 g SO4-S/L day, and a hydraulic retention time (HRT) of 2 days, a sulfate removal efficiency greater than 93% could be achieved. When the filter was operated under COD (chemical oxygen demand)/S from 10/1 to 5/1 and HRTs of 2, 1 and 0.5 days, sulfate removal efficiency was between 98.1 and 70.9%. It is believed that protection by the immobilized cell structure caused the microbial cells in the filter to tolerate higher dissolved sulfide (447.8 mg/L) and total sulfide (940.3 mg/L) levels, allowing a much higher biomass concentration (13.2-13.5 g VSS/L) to be reached.

Bacteria, Anaerobic↗