Facultative Anaerobic Species Research Articles

Use of Bacteria in Cancer Therapy.

Cancer is one of the most dreaded diseases in humans and most common cause of death in twenty-first century. New cancer therapies are urgently required because of the existing pharmacological side effects of the conventional chemotherapy, radiation, or surgery. Newer modalities such as cancer vaccines and biological therapies are proving very helpful in the treatment of cancer along with the conventional therapies. The success of these novel cancer therapies is attributed to their lesser toxicity and the specific killing of the cancer cells. Bacterial therapy for cancer has been recognized a century ago. Live, attenuated, or genetically modified obligate or facultative anaerobic bacterial species exhibit the inherent property of colonizing the tumors and are capable of multiplying selectively inside the tumors, thereby inhibiting cancerous growths. The bacteria and their spores are used in the target specific therapies, delivering the prodrugs and the various proteins to the tumors. Albeit bacterial treatment of cancer is providing new perspective in the treatment of disease, the use of microorganisms to target tumors has certain confinements. The biosafety, genetic instability and the confounded interaction of the bacteria with treatment drugs, requires the more noteworthy consideration regarding the use of this novel treatment in the cancer treatment.

The Hoopoe's Uropygial Gland Hosts a Bacterial Community Influenced by the Living Conditions of the Bird.

Molecular methods have revealed that symbiotic systems involving bacteria are mostly based on whole bacterial communities. Bacterial diversity in hoopoe uropygial gland secretion is known to be mainly composed of certain strains of enterococci, but this conclusion is based solely on culture-dependent techniques. This study, by using culture-independent techniques (based on the 16S rDNA and the ribosomal intergenic spacer region) shows that the bacterial community in the uropygial gland secretion is more complex than previously thought and its composition is affected by the living conditions of the bird. Besides the known enterococci, the uropygial gland hosts other facultative anaerobic species and several obligated anaerobic species (mostly clostridia). The bacterial assemblage of this community was largely invariable among study individuals, although differences were detected between captive and wild female hoopoes, with some strains showing significantly higher prevalence in wild birds. These results alter previous views on the hoopoe-bacteria symbiosis and open a new window to further explore this system, delving into the possible sources of symbiotic bacteria (e.g. nest environments, digestive tract, winter quarters) or the possible functions of different bacterial groups in different contexts of parasitism or predation of their hoopoe host.

Structure activity characterization of Bordetella petrii lipid A, from environment to human isolates.

Bordetella petrii, a facultative anaerobic species, is the only known member of the Bordetella genus with environmental origin. However it was also recently isolated from humans. The structures of the B.petrii lipid A moieties of the endotoxins were characterized here for the first time for an environmental strain and compared to that of human isolates. Characterization was achieved using chemical analyses, gas chromatography-mass spectrometry, and Matrix Assisted Laser Desorption Ionisation mass spectrometry. The analyses revealed that the different lipid A structures contain a common bisphosphorylated β-(1→6)-linked d-glucosamine disaccharide with hydroxytetradecanoic acid in amide as well at the C-3' in ester linkages. Similar to Bordetella pertussis and Bordetella bronchiseptica lipids A, the hydroxytetradecanoic acid at the C-2' position was substituted by tetradecanoic acid. Unlike B.pertussis, the hydroxytetradecanoic acid at the C-2 position was substituted with either 12:0 or 14:0 and/or their 2-OH forms. Depending on the environmental or human origin the structures differed in the length and degree of fatty acid acylation and impacted the IL-6 and TNF-α inflammatory responses tested. In one isolate we showed the presence at the C-3 position of the short-chain 10:0(3-OH), which according to our previous analyses is more characteristic of the human pathogens in the genus like B.pertussis and Bordetella parapertussis.

Viable bacteria associated with red blood cells and plasma in freshly drawn blood donations.

ObjectivesInfection remains a leading cause of post-transfusion mortality and morbidity. Bacterial contamination is, however, detected in less than 0.1% of blood units tested. The aim of the study was to identify viable bacteria in standard blood-pack units, with particular focus on bacteria from the oral cavity, and to determine the distribution of bacteria revealed in plasma and in the red blood cell (RBC)-fraction.DesignCross-sectional study. Blood were separated into plasma and RBC-suspensions, which were incubated anaerobically or aerobically for 7 days on trypticase soy blood agar (TSA) or blue lactose plates. For identification colony PCR was performed using primers targeting 16S rDNA.SettingBlood donors attending Capital Region Blood Bank, Copenhagen University Hospital, Rigshospitalet, Hvidovre, Denmark, October 29th to December 10th 2013.Participants60 donors (≥50 years old), self-reported medically healthy.ResultsBacterial growth was observed on plates inoculated with plasma or RBCs from 62% of the blood donations. Growth was evident in 21 (35%) of 60 RBC-fractions and in 32 (53%) of 60 plasma-fractions versus 8 of 60 negative controls (p = 0.005 and p = 2.6x10-6, respectively). Propionibacterium acnes was found in 23% of the donations, and Staphylococcus epidermidis in 38%. The majority of bacteria identified in the present study were either facultative anaerobic (59.5%) or anaerobic (27.8%) species, which are not likely to be detected during current routine screening.ConclusionsViable bacteria are present in blood from donors self-reported as medically healthy, indicating that conventional test systems employed by blood banks insufficiently detect bacteria in plasma. Further investigation is needed to determine whether routine testing for anaerobic bacteria and testing of RBC-fractions for adherent bacteria should be recommended.

Bacteriological evaluation of tonsillar microbial flora according to age and tonsillar size in recurrent tonsillitis

Although numerous studies based on the bacteriology of the tonsil have been carried out, none of them analyzed the variation of tonsillar flora with respect to both age and tonsillar size. The purpose of this study was to isolate the facultative and obligate anaerobes both from the surface and the core of tonsils in recurrent tonsillitis as well as to analyze the variation of isolated bacterial strains according to age and tonsillar size. A prospective study was performed on 111 patients who underwent tonsillectomy. We analyzed the differences between the bacterial pathogens in recurrent tonsillitis and semi-growth estimates with regard to age and tonsillar grade. Among 111 cases, 604 bacterial strains of 21 different from the tonsil superficial and core were isolated. The most common facultative anaerobic species isolated from the surface and core were Coagulase-negative staphylococci, Alpha-hemolytic streptococci and Diphtheroid bacilli in all subgroups except patients below 8years old. The most commonly obligate anaerobic species isolated from the core were Propionibacterium acnes, Prevotella melaninogenica and Peptostreptococcus anaerobius. We found no significant difference in the cultured bacteria with respect to age and tonsillar size. The study subgroups did not differ in the occurrence of semiquantitative growth estimates of 3-4+. Our study demonstrates that there is polymicrobial aerobic and anaerobic flora in tonsils with regardless of patient's age and tonsillar size. This polymicrobial spectrum of bacteria may contribute to recurrence and to the failure of conservative treatment of these cases and therefore leads to surgical therapy.

The effects of stress hormones on growth of selected periodontitis related bacteria

The focus of this study was to examine in vitro the effects of stress hormones (catecholamines: epinephrine, norepinephrine, dopamine and hydrocortisone: cortisol) on the growth of four anaerobic species of periodontitis-related bacteria (Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia and Tannerella forsythia) and one facultative anaerobic species (Eikenella corrodens).Bacterial growth was determined by two different methods: fluorescence in situ hybridization (FISH), and the viable count by culture method. To simulate stress, each single strain was grown in a special growth medium with three different concentrations of each hormone, using an anaerobic chamber at 37 °C.Growth of F. nucleatum increased in the presence of all stress hormones. Growth of P. gingivalis was not significantly influenced by any hormone. Growth of P. intermedia and E. corrodens was inhibited by almost all stress hormones tested. Both methods of analysis revealed that the highest concentrations of norepinephrine and cortisol increased the growth of T. forsythia. Different hormones have a different effect on the growth of periodontitis-related bacteria in vitro. It appears that bacterial viability is more strongly influenced than is bacterial metabolic activity. The growth of F. nucleatum particularly and partially of T. forsythia is increased by several stress hormones and may have an additional negative impact on periodontal disease.

Draft Genome Sequence of Weissella koreensis KCTC 3621T

Weissella koreensis is a Gram-positive, rod-shaped, nonmotile, and facultative anaerobic species belonging to the lactic acid bacteria (LAB). The members of this species have been repeatedly isolated from kimchi (a traditional Korean fermented food) and are known for their beneficial effects on human and animal intestinal microflora through producing various clinically important amino acids such as γ-aminobutyric acid and ornithine. Here we report the genome sequence of the type strain of W. koreensis (KCTC 3621(T)) to provide taxonomic and functional insights into the species.

Aggregatibacter actinomycetemcomitans targets NLRP3 and NLRP6 inflammasome expression in human mononuclear leukocytes

Periodontitis is an inflammatory condition that destroys the tooth-supporting tissues, as a result of local bacterial infection. Aggregatibacter actinomycetemcomitans is a Gram-negative facultative anaerobic species, highly associated with aggressive periodontitis. Periodontal inflammation is dominated by cytokines of the Interleukin (IL)-1 family. Prior to their secretion by mononuclear cells, IL-1 cytokines are processed by intracellular protein complexes, known as “inflammasomes”, which can sense the bacterial challenge. The aim of this study was to investigate which inflammasomes are regulated in mononuclear cells in response to A. actinomycetemcomitans. The D7SS strain and its derivative leukotoxin and cytolethal distending toxin knock-out mutant strains were used to infect human mononuclear cells at a 1:10 cell: bacteria ratio, for 3h. The expression of various inflammasome components in the cells was investigated by TaqMan quantitative real-time polymerase chain reaction (qPCR). The expressions of NOD-like receptor protein (NLRP)1, NLRP2 and Absent In Melanoma (AIM)2 inflammasome sensors, as well as their effector Caspase-1 were not affected. However, NLRP3 was up-regulated, while NLRP6 was down-regulated. This effect was not dependent on the leukotoxin or the cytolethal distending toxin, as demonstrated by the use of specific gene knock-out mutant strains. IL-1β and IL-18 expressions were also up-regulated by the bacterial challenge. In conclusion, A. actinomycetemcomitans enhances NLRP3 and reduces NLRP6 inflammasome expression, irrespective of its major virulence factors, confirming the high pathogenic profile of this species, and providing further insights to the mechanisms of periodontal inflammation.

Bovine Intestinal Bacteria Inactivate and Degrade Ceftiofur and Ceftriaxone with Multiple β-Lactamases

The veterinary cephalosporin drug ceftiofur is rapidly degraded in the bovine intestinal tract. A cylinder-plate assay was used to detect microbiologically active ceftiofur, and high-performance liquid chromatography-mass spectrometry analysis was used to quantify the amount of ceftiofur remaining after incubation with bovine intestinal anaerobic bacteria, which were isolated from colon contents or feces from 8 cattle. Ninety-six percent of the isolates were able to inactivate ceftiofur to some degree, and 54% actually degraded the drug. None of 9 fungal isolates inactivated or degraded ceftiofur. Facultative and obligate anaerobic bacterial species that inactivated or degraded ceftiofur were identified with Vitek and Biolog systems, respectively. A subset of ceftiofur degraders also degraded the chemically similar drug ceftriaxone. Most of the species of bacteria that degraded ceftiofur belonged to the genera Bacillus and Bacteroides. PCR analysis of bacterial DNA detected specific β-lactamase genes. Bacillus cereus and B. mycoides isolates produced extended-spectrum β-lactamases and metallo-β-lactamases. Seven isolates of Bacteroides spp. produced multiple β-lactamases, including possibly CepA, and metallo-β-lactamases. Isolates of Eubacterium biforme, Bifidobacterium breve, and several Clostridium spp. also produced ceftiofur-degrading β-lactamases. An agar gel overlay technique on isoelectric focusing separations of bacterial lysates showed that β-lactamase enzymes were sufficient to degrade ceftiofur. These results suggest that ceftiofur is inactivated nonenzymatically and degraded enzymatically by multiple β-lactamases from bacteria in the large intestines of cattle.

The Microfloral Analysis of Secondary Caries Biofilm around Class I and Class II Composite and Amalgam Fillings

BackgroundSecondary caries is responsible for 60 percent of all replacement restorations in the typical dental practice. The diversity of the bacterial sources and the different types of filling materials could play a role in secondary caries. The aim of this study was to determine and compare the microbial spectrum of secondary caries biofilms around amalgam and composite resin restorations.MethodsClinical samples were collected from freshly extracted teeth diagnosed with clinical secondary caries. Samples were categorized into four groups according to the types of restoration materials and the classification of the cavity. Biofilms were harvested from the tooth-restoration interface using a dental explorer and after dilution were incubated on special agars. The bacteria were identified using the biochemical appraisal system. Statistical calculations were carried out using SPSS11.5 software to analyze the prevalence of the bacteria involved in secondary caries.ResultsSamples from a total of four groups were collected: two groups were collected from amalgam restorations, each had 21 samples from both Class I and Class II caries; and the other two groups were from composite resin restorations, each had 13 samples from both class I and class II caries. Our results showed: (1) Anaerobic species were dominant in both restoration materials. (2) In terms of the types of individual bacteria, no significant differences were found among the four groups according to the geometric mean of the detected bacteria (P > 0.05). However, there were significant differences among the detected bacteria within each group (P < 0.05). The composition of each bacterium had no statistical difference among the four groups (P > 0.05), but showed significant differences among the detected bacteria in each group (P < 0.05). (3) Among the four groups, there were no significant differences for the detection rate of each bacterium (P > 0.05), however, the detection rate of each bacterium within each group was statistically different among the detected bacteria (P < 0.05).ConclusionsThe proportion of obligatory anaerobic species was much greater than the facultative anaerobic species in the biofilm of secondary caries. Statistically, the materials of restoration and the location of secondary caries did not show any significant effects on the composition of the microflora.

A first report on the microbial colonisation of the equine oesophagus

Based on cryo-SEM, standard and high resolution TEM, glycoconjugate histochemistry, and microbiological differentiation, the present study demonstrates the colonisation of the epithelium of the equine oesophagus with microorganisms. As particularly apparent using cryo-SEM to illustrate natural conditions, the present microbiota were clearly dominated by bacteria, forming a one-layer system, as attached to and embedded in concentrated mannose/mannan substances covering the outer stratum corneal cells. Bacterial numbers ranged from 5600 to 7200 per mm 2 in the central part of the oesophagus, the number of fungi was less than 1% of the amount of bacteria. The compact stratum corneal cells showed numerous short protrusions sometimes as part of desmosomal contacts, but mainly projecting into distinct intercellular spaces, containing a mixture of acid and neutral glycoconjugates. The outermost corneal cells exhibited intact mitochondria and cytoplasmic vesicles, and a number of short cell processes toward the oesophageal lumen; i.e. into the glycoconjugate layers on the surface of the oesophagus. The diverse spectrum of bacteria found indicated a permanent mucosal flora, predominated by facultative and obligate anaerobic species. The genera isolated most frequently and in highest numbers included streptococci, Prevotella spp., Fusobacterium spp. and Actinobacillus equuli. Only two groups of Enterobacteriaceae ( Escherichia coli, Pantoea spp.) were regularly found and their abundance was lower than that of the other bacterial groups mentioned above. Yeasts were very rarely identified as the typically present fungi.

Decomposition and mineralization of aquatic humic substances (AHS) in treating landfill leachate using the Anammox process

Aquatic humic substance (AHS) in landfill leachate is resistant to biodegradation, especially in anaerobic habitats. However, we reported here for the first time that AHS was completely biodegraded with the anaerobic ammonium oxidation (Anammox) process. As a result of aromatic-ring cleavage and mineralization, carboxylic and aliphatic organics were significantly produced, the contents of chromophores such as quinoid and ketone were remarkably decreased to decolorize the leachate, and produced carbon dioxide along with increasing carbonate was clearly presented. Of the degraded AHS of 137mgL−1, 51mgL−1 was owed to the oxidation with sulfate of 76mgL−1 as electron acceptor, and the rest to other metabolic mechanism. Isolation and identification of heterotrophic bacteria revealed a diversified consortium comprising four facultative anaerobic species, Bacillus sp., Paenibacillus sp., Bacteroides sp. and Staphylococcus sp., without sulfate-reducing bacteria detected. Their contribution to AHS biodegradation and sulfate reduction under the special conditions with high oxidization–reduction potential and insufficient electron acceptors has not been known yet. Further work is underway to investigate their properties and respective duties in the consortium.

Physiological heterogeneity of suspended microbial aggregates

Suspended microbial aggregates, which are always in dynamic equilibrium with suspended cells and cells attached to surface, are very common structures in natural and engineering environmental systems. To study and design physiologically diverse suspended microbial aggregates the physiological classification of chemotrophic prokaryotes in 12 groups formed by four evolutionary periods (fermenting, anaerobic respiring, microaerophilic and facultative aerobic, aerobic prokaryotes) and three parallel lines (Gram-negative, Gram-positive Eubacteria, and Archaea) could be used. This type of physiological heterogeneity has been studied in microbial granules using fluorescence in situ hybridization, identification of 16S rRNA genes, and conferring the physiological properties from the description of the species. In spherical granules with diameter of 2.4 mm cells of aerobes were spread to the depth 0.55 mm below surface (85% of granule volume), facultative anaerobes dominated between the depths 0.55 mm and 0.85 mm (13% of granule volume), and anaerobes were concentrated at the depths from 0.85 to 1.0 mm (2% of granule volume). Percentages of aerobic, facultative anaerobic, and anaerobic species in granules, identified by 16S rRNA gene sequencing, were 69%, 9%, and 2% of total number of bacterial clones, respectively. Another type of physiological heterogeneity on the cellular level was due to the changes of cell physiological status during cell cycle. This type of heterogeneity has been studied in the populations of Escherichia coli, Bacillus megaterium, Saccharomyces cerevisiae, and Candida tropicalis. A significant proportion of cells from the exponential phase were killed after 10 min treatment with 1% solution of allyl alcohol, which specifically kills cells with high activity of alcohol dehydrogenase (ADH). However, there was no such effect in starved cell population. Percentage of cells with high activity ADH in microbial population can be used to monitor its physiological status. Physiological diversity of ecosystem may be due to mechanical mixing of cells from the different inflows. An example of such system is an ecosystem of aeration tank in municipal wastewater treatment plant. This ecosystem contains a mechanical mixture of dead anaerobic and live aerobic bacteria as well as attached and suspended cell aggregates supplied from anaerobic digester, raw sewage, and settling tank.

Role of Serratia marcescens ACE2 on diesel degradation and its influence on corrosion

A facultative anaerobic species Serratia marcescens ACE2 isolated from the corrosion products of diesel transporting pipeline in North West, India was identified by 16S rDNA sequence analysis. The role of Serratia marcesens ACE2 on biodegradation of diesel and its influence on the corrosion of API 5LX steel has been elucidated. The degrading strain ACE2 is involved in the process of corrosion of steel API 5LX and also utilizes the diesel as an organic source. The quantitative biodegradation efficiency (BE) of diesel was 58%, calculated by gas-chromatography-mass spectrum analysis. On the basis of gas-chromatography-mass spectrum (GC-MS), Fourier Transform infrared spectroscopy (FTIR) and X-ray diffractometer (XRD), the involvement of Serratia marcescens on degradation and corrosion has been investigated. This basic study will be useful for the development of new approaches for detection, monitoring and control of microbial corrosion.

Caries‐related plaque microcosm biofilms developed in microplates

In vivo dental plaque biofilms consist of complex communities of oral bacteria that are a challenge to replicate in vitro. The aim of this investigation was to establish human dental plaque microcosms in microplates to reflect conditions that are relevant to dental caries. Microcosm plaque biofilms were initiated from the saliva of two different donors, grown for up to 10 days in 24-welled microplates on Thermanox coverslips in various types of artificial saliva with and without sucrose, which were replaced daily. Microbiota composition of 40 species associated with oral health and dental caries was monitored in the plaques using Checkerboard DNA-DNA hybridization analysis. pH was measured as an indicator of cariogenic potential. The composition of the saliva inocula was different, and yielded plaque microcosms with different composition and growth responses to sucrose. Artificial saliva type and presence of sucrose, and the resulting growth and pH conditions, modified the growth of individual species and hence the ecological profile of the microplate plaques during development. Complex population shifts were observed during development, and older plaques comprised predominantly facultative anaerobic species. Sucrose supplementation limited the decline of Streptococci over time but did not increase the abundance of mutans Streptococci. Sucrose at 0.15% increased levels of caries-associated species including Lactobacillus fermentum, Lactobacillus acidophilus and Actinomyces gerensceriae; these were further increased with sucrose at 0.5%, in addition to Actinomyces israelii, Rothia dentocariosa and Capnocytophaga gingivalis. The microplate plaques demonstrated complex community dynamics that appeared to reflect the maturation of natural plaques, and sucrose induced a cariogenic plaque composition and pH.

Biodegradation and corrosion behaviour of Serratia marcescens ACE2 isolated from an Indian diesel-transporting pipeline

A facultative anaerobic species Serratia marcescens ACE2 isolated from the corrosion products of a diesel-transporting pipeline in North West India was identified by 16S rDNA sequence analysis. The role of Serratia marcesens ACE2 on biodegradation of commercial corrosion inhibitor (CCI) and its influence on the corrosion of API 5LX steel has been enlightened. The degrading strain ACE2 is involved in the process of corrosion of steel API 5LX and also utilizes the inhibitor as organic source. The quantitative biodegradation efficiency of corrosion inhibitor was 58%, which was calculated by gas chromatography mass spectrum analysis. The effect of CCI on the growth of bacteria and its corrosion inhibition efficiency were investigated. Additionally, the role of this bacterium in corrosion of steel has been investigated by powder X-ray diffractometer (XRD) and scanning electron microscope studies. The presence of high-intensity ferric oxides and manganese oxides noticed from the XRD indicates that ACE2 enhances the corrosion process in presence of inhibitor as a carbon source. This basic study will be useful for the development of new approaches for the detection, monitoring and control of microbial corrosion in petroleum product pipelines.

Characterization of the xylanases from the new isolated thermophilic xylan-degrading Bacillus thermoleovorans strain K-3d and Bacillus flavothermus strain LB3A.

Three strictly aerobic strains (K-1, K-3d and K-4) were isolated from a hot-spring in Kobe, Japan, and a facultative anaerobic strain LB3A was isolated from sediments collected from the alkaline Lake Bogoria, Kenya. All strains were thermophilic and capable of growth on xylan. On the basis of morphological, physiological and phylogenetic studies the new aerobic isolates resemble the thermophilic species Bacillus thermoleovorans while the facultative anaerobic isolate LB3A resembles the facultative anaerobic thermophilic species Bacillus flavothermus. When grown on xylan as sole carbon source, all isolates produce thermoactive xylanases, Xylanases from strains K-3d and LB3A are active at temperatures between 40 and 90 degrees C and pH values between 5.0 and 9.0. Applying SDS-PAGE the crude xylanase complex of isolate K-3d was shown to be composed of two active bands, with molecular masses of 40 and 69 kDa. The crude xylanase complex of isolate LB3A, on the other hand, is composed of at least four activity bands with molecular masses ranging from 80 to 130 kDa. Due to the product pattern of xylan hydrolysis both enzymes are classified as endoxylanases. The xylanolytic enzyme system of isolate K-3d produces xylotriose, xylotetraose and larger xylooligosacharides, whereas the xylanases from isolate LB3A release xylotetraose as the major product of hydrolysis.

Microbiological evaluation of endodontic files after cleaning and steam sterilization procedures

Infection control procedures are essential for modern dental practice and they are continually evolving to meet the dental profession's high standards. The present study evaluated the efficacy of two cleaning procedures to reduce bacterial numbers on endodontic files, and evaluated the effect of biological debris on the subsequent sterilization of files. Stainless steel and nickel-titanium (NiTi) files were examined upon removal from the manufacturer's packaging, after instrumentation in root canals of human teeth inoculated with a broth containing two anaerobic species and one facultative anaerobic species of bacteria, and after instrumentation and cleaning with either an ultrasonic bath or a thermal disinfector. For each file, the bacterial numbers were quantified using routine microbiological techniques in an anaerobic chamber. No bacteria were detected from files direct from their packets. The size, taper and type of file did not affect the ability of either of the cleaning procedures to reduce bacterial numbers. However, an absence of bacteria was more likely when files were cleaned in the thermal disinfector. No bacteria were detected from files that were-subjected to steam sterilization irrespective of the type of prior cleaning procedure. Steam sterilization eliminated all bacteria from the endodontic files irrespective of the presence of biological debris. The majority of bacteria were eliminated from endodontic files after either ultrasonic cleaning or using a thermal disinfector.

Microorganisms from canals of root-filled teeth with periapical lesions.

The objective of the present study was to identify the microbial flora within root canals of teeth with failed root-canal treatment and to determine the association of the various species with clinical features. Sixty root-filled teeth with persisting periapical lesions were selected for this study. During nonsurgical endodontic re-treatment, the root-filling material was removed and the canals were sampled. Microbial sampling, isolation and species determination were performed using advanced microbiological techniques for anaerobic species. The association of microbiological findings with clinical features was investigated. Microorganisms were recovered from 51 teeth. In most cases, one or two strains per canal were found. Of the microbial species isolated, 57.4% were facultative anaerobic species and 83.3% Gram-positive microorganisms. Enterococcus faecalis was the most frequently recovered bacterial species. Obligate anaerobes accounted for 42.6% of the species and the most frequently isolated genera was Peptostreptococcus. which was associated with clinical symptoms (P < 0.01). Significant associations were also observed between: (a) pain or history of pain and polymicrobial infections or anaerobes (P < 0.05): (b) tenderness to percussion and Prevotella intermedia/P. nigrescens (P < 0.05); (c) sinus and Streptococcus spp. (P < 0.001) or Actinomyces spp. (P < 0.01); (d) coronally unsealed teeth and Streptococcus spp. or Candida spp. (both with P < 0.01). The microbial flora in canals after failure of root-canal treatment were limited to a small number of predominantly Gram-positive microbial species. Facultative anaerobes, especially E. faecalis, were the most commonly isolated microorganisms, however, polymicrobial infections and obligate anaerobes were frequently found in canals of symptomatic root-filled teeth.

Galactooligosaccharide in combination with Bifidobacterium and Bacteroides affects the population of Clostridium perfringens in the intestine of gnotobiotic mice

The purpose of this study was to determine the effect of 5% galactooligosaccharide (GOS) on the population of Clostridium perfringens during the development of primary intestinal microflora in gnotobiotic mice. Bifidobacterium breve markedly increased and C. perfringens markedly decreased in populations in the feces of the GOS-fed animals, but not in the controls. Bacteroides vulgatus in association with B. breve depressed the C. perfringens growth with the aid of dietary GOS. The depression of C. perfringens growth, however, depended on the timing of its colonization. This study indicates that Lactobacillus salivarius colonization is stimulated by dietary GOS and that growth of C. perfringens is suppressed by dietary GOS in the combination with anaerobic bacteria such as Bifidobacterium and Bacteroides and facultative anaerobic species during the early development of the bowel microflora, provided bacterial species colonize the gut in appropriate timing.