Chemokines expression during Leptospira interrogans serovar Copenhageni infection in resistant BALB/c and susceptible C3H/HeJ mice

The role of innate immune responses in protection against leptospirosis remains unclear. We examined the expression of the chemokines CCL2/JE (MCP-1), CCL3/MIP-1 alpha (MIP-1 alpha) and CXCL1/KC (IL-8) regarding resistance and susceptibility to leptospirosis in experimental mice models BALB/c and C3H/HeJ, respectively. A virulent strain of Leptospira interrogans serovar Copenhageni was used in this study. Twenty-five animals of each mouse strain of C3H/HeJ and BALB/c, were infected intraperitoneally with 106 cells. Five un-infected animals of each strain were kept as control. Mortality of C3H/HeJ mouse was observed while BALB/c mice were asymptomatic. The presence of leptospire DNA in tissues of infected animals was demonstrated by PCR. Chemokines were measured in serum, spleen, liver, kidney and lung of both strains of animals using immunoenzymatic assay (ELISA). Elevations in the levels of chemokines MCP-1 and IL-8 occurred in all organs and sera of C3H/HeJ and BALB/c infected mice. The levels of MIP-1 alpha were lower when compared to MCP-1 and IL-8 in all analyzed organs, with a slight increase in liver and kidney. Our results indicate that the expression of inflammatory mediators can vary greatly, depending on the tissue and mouse strains. It is possible that the resistance to Leptospira can be partially correlated to the increase of MIP-1 alpha observed in BALB/c mice, while an increasing and a sustained expression of MCP-1 and IL-8 in the lungs of C3H/HeJ mice can be correlated to the severity and progression of leptospirosis. (C) 2009 Elsevier Ltd. All rights reserved.

In vitro evidence for immune evasion activity by human plasmin associated to pathogenic Leptospira interrogans

Leptospirosis is a widespread re-emerging zoonosis of human and veterinary concern. It has been shown that virulent leptospires protect themselves against the host`s innate immune system, a strategy that allows the bacteria to reach immunologically safe environments. Although extensive studies on host pathogen interactions have been performed, little is known on how leptospires deal with host immune attack. In a previous work, we demonstrated the ability of leptospires to bind human plasminogen (PLC), that after treatment with activators, conferred plasmin (PLA) activity on the bacteria surface. In this study, we show that the PLA activity associated to the outer surface of Leptospira could interfere with the host immune attack by conferring some evasion advantage during infection. We demonstrate that PLA-coated leptospires interfere with complement Ob and IgG depositions on the bacterial surface, probably through the degradation of these components, thus diminishing opsonization process. Similar decrease on the deposition was observed when normal and immune sera from patients diagnosed with leptospirosis were employed as a source of IgG. We believe that decreasing opsonization by PLA generation might be an important aspect of the leptospiral immune escape strategy and survival. To our knowledge, this is the first proteolytic activity of plasmin associated-Leptospira related to anti-opsonic properties reported to date. (C) 2011 Elsevier Ltd. All rights reserved.

Wide Dispersal and Possible Multiple Origins of Low-Copy-Number Plasmids in Rickettsia Species Associated with Blood-Feeding Arthropods

Plasmids are mobile genetic elements of bacteria that can impart important adaptive traits, such as increased virulence or antibiotic resistance. We report the existence of plasmids in Rickettsia (Rickettsiales; Rickettsiaceae) species, including Rickettsia akari, ""Candidatus Rickettsia amblyommii,"" R. bellii, R. rhipicephali, and REIS, the rickettsial endosymbiont of Ixodes scapularis. All of the rickettsiae were isolated from humans or North and South American ticks. R. parkeri isolates from both continents did not possess plasmids. We have now demonstrated plasmids in nearly all Rickettsia species that we have surveyed from three continents, which represent three of the four major proposed phylogenetic groups associated with blood-feeding arthropods. Gel-based evidence consistent with the existence of multiple plasmids in some species was confirmed by cloning plasmids with very different sequences from each of two ""Ca. Rickettsia amblyommii"" isolates. Phylogenetic analysis of rickettsial ParA plasmid partitioning proteins indicated multiple parA gene origins and plasmid incompatibility groups, consistent with possible multiple plasmid origins. Phylogenetic analysis of potentially host-adaptive rickettsial small heat shock proteins showed that hsp2 genes were plasmid specific and that hsp1 genes, found only on plasmids of ""Ca. Rickettsia amblyommii,"" R. felis, R. monacensis, and R. peacockii, were probably acquired independently of the hsp2 genes. Plasmid copy numbers in seven Rickettsia species ranged from 2.4 to 9.2 per chromosomal equivalent, as determined by real-time quantitative PCR. Plasmids may be of significance in rickettsial evolution and epidemiology by conferring genetic plasticity and host-adaptive traits via horizontal gene transfer that counteracts the reductive genome evolution typical of obligate intracellular bacteria.

Productive performance, nutrient digestion and metabolism of Holstein (Bos taurus) and Nellore (Bos taurus indicus) cattle and Mediterranean Buffaloes (Bubalis bubalis) fed with corn-silage based diets

The objective of this study was to evaluate the productive performance, nutrients digestion and metabolism of three different genetic groups fed with the same diet based on corn silage. 30 heifers in growth were used of three groups of cattle, the following: Nellore (Bos taurus indicus) (n = 10), Holstein (Bos taurus taurus) (n = 10), and Mediterranean buffaloes (Bubalis bubalis) (n = 10). The animals were fed in groups and received the same experimental diet composed of corn silage and concentrate for growing heifers. In the evaluation of animals the performance, consumption and total apparent digestibility of dry matter and nutrients with the aid of internal markers (chromic oxide) and external (iADF), rumen fermentation, excretion of purine derivatives, nitrogen balance and blood metabolites were measured. No differences were observed in animal performance. There were differences in nutrient intake and apparent digestibility of dry matter and nutrients in different groups of cattle. The concentration of ammonia nitrogen (NH3-N) and short chain fatty acids (SCFA) in the rumen were higher and lower, respectively, for the group of buffaloes in relation to other experimental groups evaluated. When considering the excretion of total purine derivatives, buffaloes showed the lowest value compared to other genetic groups evaluated; about 61.76% of the total genetic group Nellore and 57.62% of the total genetic group Holstein with an average of 33.67 mmol/day. For the buffaloes, the excretion of xanthine and hypoxanthine observed was of 5.11% of total purine derivatives. There was a better nitrogen balance (g/day) for groups of Holstein heifers and Nellore in relation to the group of buffalo heifers. There were differences in the concentrations of urea and urea nitrogen in serum and liver enzymes where the buffaloes had higher values in relation at the bovines. There is a great metabolic diversity among the experimental groups evaluated and it was more exacerbated among buffaloes and bovines, when submitted to the same diet and same management conditions. (C) 2011 Elsevier B.V. All rights reserved.

Evaluation of somatic cell count thresholds to detect subclinical mastitis in Gyr cows

The objectives of this study were (1) to determine the sensitivity (Se) and specificity (Sp) of somatic cell count (SCC) thresholds to identify subclinical mastitis in Gyr cows caused by major and minor pathogens; (2) to study the effects of month of sampling, rear or front mammary quarters, herd, intramammary infection (IMI), and bacterial species on SCC at quarter level; and (3) to describe the prevalence of IMI in Gyr cows in commercial dairy herds. In total, 221 lactating Gyr cows from 3 commercial dairy farms were selected. Milk samples were collected from individual quarters once a month for 1 yr from all lactating cows for SCC and bacteriological analysis. Mammary quarters were considered the experimental units and the SCC results were log(10)-transformed. Four SCC thresholds (100, 200, 300 and 400 x 10(3) cells/mL) were used to determine Se and Sp to identify infected mammary quarters. The overall prevalence of IMI in quarter milk samples of Gyr cows was 49.8%, and the prevalence of minor pathogens was higher (31.9%) than that of major pathogens (17.8%). Quarter samples with microbial isolation presented higher SCC compared with negative samples. Sensitivity and Sp of selected SCC thresholds varied according to the group of pathogen (major and minor) involved in the IMI definition. Sensitivity increased and Sp decreased when mammary quarters with only major pathogens isolation were considered positive. The use of a single SCC analysis to classify quarters as uninfected or infected in Gyr cows may not be a useful test for this breed because Se and Sp of SCC at the studied thresholds were low. The occurrence of IMI and the bacterial species are the main factors responsible for SCC variation in mammary quarters of Gyr cows. Milk samples with major pathogens isolation elicited higher SCC than those with minor pathogens.

In vitro analysis of inhibitory effects of the antibacterial monomer MDPB-containing restorations on the progression of secondary root caries

Objective: This study aimed to analyze in vitro inhibitory effects of restorative materials containing the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) on the formation of artificial secondary root caries lesions. Methods: Class V cavities (2 mm x 2 mm) were prepared in 75 human root fragments. Specimens were randomly divided into five groups (n = 15 fragments per group) and restored as follows: (I) MDPB-free adhesive system + MDPB-free composite (negative control); (II) resin modified glass ionomer (RM-GIC; positive control); (III) MDPB-free adhesive system + MDPB-containing composite (2.83% MDPB); (IV) MDPB-containing adhesive system + MDPB-free composite; M MDPB-containing adhesive system + MDPB-containing composite. Artificial secondary root caries lesions were produced by a biological artificial caries challenge. The restored specimens were immersed into a culture medium containing Streptococcus mutans and sucrose for 15 days. Histological slices (80 +/- 20 mu m) of the specimens were used for measuring the mean depths of the artificial lesions produced in both margins of the restorations using polarized light microscopy. Results were expressed in percentage related to the mean depth of the negative control, considered 100%. Data were compared by ANOVA followed by the Tukey`s test (p <= 0.05). Results: The depths of lesions adjacent to cavities filled with RM-GIC (GII; 85.17 +/- 15.2%) were significantly (p < 0.01) shallower than those adjacent to restorations with MDPB-free composite (GI; 100.00 +/- 10.04%), despite the presence of MDPB in the adhesive system (GIV; 101.95 +/- 21.32%). The depths of lesions adjacent to cavities restored with MDPB-containing composite (GIII; 82.68 +/- 12.81% and GV; 85.65 +/- 15.42%), despite the adhesive system used, were similar to those of RM-GIC (GII). Mean lesions depths in these groups decreased from 13% (GV) to 17% (GIII) in relation to the negative control (GI). Conclusions: MDPB-containing composite inhibits the progression of artificial secondary root caries lesions regardless of adhesive systems. (C) 2009 Elsevier Ltd. All rights reserved.

In vitro antimicrobial activity of Caesalpinia ferrea Martius fruits against oral pathogens

Aim: In the Amazon region of Brazil, the fruits of Caesalpinia ferrea Martius (Brazilian ironwood) are widely used as an antimicrobial and healing medicine in many situations including oral infections. This study aimed to evaluate the antimicrobial activity of Caesalpinia ferrea Martius fruit extract against oral pathogens. Materials and methods: Polyphenols estimation and spectral analysis ((1)H NMR) of the methanol extract were carried out. The microorganisms Candida albicans, Streptococcus mutans, Streptococcus salivarius, Streptococcus oralis and Lactobacillus casei were tested using the microdilution method for planktonic cells (MIC) and a multispecies biofilm model. Chlorhexidine was used as positive control. Results: Polyphenols in the extract were estimated at 7.3% and (1)H NMR analysis revealed hydroxy phenols and methoxilated compounds. MIC values for Candida albicans, Streptococcus mutans, Streptococcus salivarius, Streptococcus oralis and Lactobacillus casei were 25.0, 40.0, 66.0, 100.0, 66.0 mu g/mL, respectively. For the biofilm assay, chlorhexidine and plant extract showed no growth at 10(-4) and 10(-5) microbial dilution, respectively. At 10-4 and 10-5 the growth values (mean +/- SD) of the negative controls (DMSO and saline solution) for Streptococcus mutans, Streptococcus sp. and Candida albicans were 8.1 +/- 0.7, 7.0 +/- 0.6 and 5.9 +/- 0.9 x 10(6) CFU, respectively. Conclusion: Caesalpinia ferrea fruit extract can inhibit in vitro growth of oral pathogens in planktonic and biofilm models supporting its use for oral infections. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Inflammatory Papillary Hyperplasia of the Palate: Quantitative Analysis of Candida albicans and its Negative Correlation with Microscopic and Demographic Aspects

Inflammatory papillary hyperplasia of the palate (IPHP) is a tissue-reactive overgrowth characterized by hyperemic mucosa with nodular or papillary appearance in the palate. The exact pathogenesis is still unclear. In this study, the presence of Candida albicans in the epithelial lining was evaluated using the indirect immunofluorescence staining technique. Strongly stained C albicans was observed only in the lesions of the IPHP group. Therefore, the detection of C albicans in almost all samples from IPHP tissue enabled a suggestion as to the microbial etiology of the disease, since the use of dental prostheses was reported. Int J Prosthodont 2011;24:235-237

State of the art etch-and-rinse adhesives

Objectives: The aim of this study was to explore the therapeutic opportunities of each step of 3-step etch-and-rinse adhesives. Methods: Etch-and-rinse adhesive systems are the oldest of the multi-generation evolution of resin bonding systems. In the 3-step version, they involve acid-etching, priming and application of a separate adhesive. Each step can accomplish multiple goals. Acid-etching, using 32-37% phosphoric acid (pH 0.1-0.4) not only simultaneously etches enamel and dentin, but the low pH kills many residual bacteria. Results: Some etchants include anti-microbial compounds such as benzalkonium chloride that also inhibits matrix metalloproteinases (MMPs) in dentin. Primers are usually water and HEMA-rich solutions that ensure complete expansion of the collagen fibril meshwork and wet the collagen with hydrophilic monomers. However, water alone can re-expand dried dentin and can also serve as a vehicle for protease inhibitors or protein cross-linking agents that may increase the durability of resin-dentin bonds. In the future, ethanol or other water-free solvents may serve as dehydrating primers that may also contain antibacterial quaternary ammonium methacrylates to inhibit dentin MMPs and increase the durability of resin-dentin bonds. The complete evaporation of solvents is nearly impossible. Significance: Manufacturers may need to optimize solvent concentrations. Solvent-free adhesives can seal resin-dentin interfaces with hydrophobic resins that may also contain fluoride and antimicrobial compounds. Etch-and-rinse adhesives produce higher resin-dentin bonds that are more durable than most 1 and 2-step adhesives. Incorporation of protease inhibitors in etchants and/or cross-linking agents in primers may increase the durability of resin-dentin bonds. The therapeutic potential of etch-and-rinse adhesives has yet to be fully exploited. (C) 2010 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Direct and transdentinal antibacterial activity of chlorhexidine

Purpose: To evaluate the antibacterial effect of different chlorhexidine (CHX) concentrations against Streptococcus mutans using the agar-diffusion method with and without human dentin discs placed between the bacteria and the test substances. Methods: For the direct application (agar-well technique), a base layer containing 15 mL of BHI agar and 300 mu L. of S. mutans inoculum (10(9) cfu/mL) was prepared in Petri dishes. Six wells per dish were made at equidistant points and immediately filled with CHX gels (0.12%, 0.2%, 1% and 2%), 35% phosphoric acid and pure natrosol (n=6 wells/substance). Paper discs soaked in sterile distilled water served as control group (n=6). For the indirect application (transdentinal diffusion), 0.2 mm- and 0.5 mm-thick human dentin discs (36 discs/thickness) had the hydraulic conductance determined, which allowed the homogeneous allocation of them to the experimental and control groups. The discs were placed at equidistant points on the Petri dishes containing BHI with the S. mutans inoculum (six discs per dish; one per substance) with the pulpal side in contact with the bacteria. In the discs treated with CHX gels, dentin surface was etched with H(3)PO(4) and rinsed with distilled water before CHX gel application for 1 minute. After both direct and indirect application, the dishes were incubated for 24 hours and the bacterial growth inhibition zones formed around the wells and dentin discs were measured. Data were analyzed statistically by the non-parametric Kruskal-Wallis and Mann-Whitney tests at 5% significance level. Results: In the direct test, all CHX concentrations presented a dose-dependent antibacterial activity against S. mutans. In the indirect test, there were statistically significant differences (P< 0.05) among all groups and the largest microbial growth inhibition zones were observed when 2% CHX was applied on 0.2 mm-thick discs (P< 0.05). It was concluded that all evaluated CHX gels exhibited both direct and transdentinal antibacterial activity against S. mutans. This effect of CHX was strongly influenced by the CHX concentration as well as the dentin barrier thickness. (Am J Dent 2010;23:255-259).

Phylogeny of the filamentous bacterium 'Nostocoida limicola' III from activated sludge

Five strains of the filamentous bacterium 'Nostocoida limicola' III were successfully isolated into pure culture from samples of activated sludge biomass from five plants in Australia. 16S rRNA gene sequence analyses showed that all isolates were members of the Planctomycetales, most closely related to Isosphaera pallida, but they differed phenotypically from this species in that they did not glide and were not thermotolerant. The ultrastructure of these 'N. limicola' III isolates was also consistent with them being Planctomycetales, in that they possessed complex intracellular membrane systems compartmentalizing the cells. However, the arrangements of these intracellular membranes differed between isolates. These data confirm that 'N. limicola' III is phylogenetically unrelated to both 'N. limicola' I and 'N. limicola' II, activated sludge filamentous bacteria which share morphological features in common with 'N. limicola' III and which have been presumed historically to be the same or very similar bacteria.

Microleakage into and from Two-Stage Implants: An In Vitro Comparative Study

Purpose: Gaps between an abutment and a dental implant are unavoidable, and microleakage may occur, leading to problems such as malodor and peri-implantitis. The aim of the present in vitro study was to investigate leakage of Staphylococcus aureus through the implant/abutment interface by the method of bacterial culture, and to compare the leakage rates of two different types of implant-abutment connections. Materials and Methods: Twenty Morse taper implants with abutments were divided into two groups: group A, which were evaluated for microleakage into the inner part of the implants, and group B, which were evaluated for microleakage from the inner part of the implants. Twenty internal-hexagon implants with abutments were also divided into two groups: group C, which were evaluated for microleakage into the inner part of the implants, and group D, which were evaluated for microleakage from the inner part of the implants. For the evaluation of leakage from the implants, the assemblies had the inner parts inoculated with S aureus, and each assembly was incubated in sterile brain heart infusion broth for 1 week. For assessment of leakage into the implants, each assembly was submerged in 4 mL S aureus culture in tubes and incubated for 2 weeks. The microleakage of the two implant connections was compared. Results: Microbial leakage occurred in all groups, and there was no statistically significant difference between groups A and C or between groups B and D. Conclusions: In vitro, S aureus leakage through the implant/abutment interface occurred with both Morse taper and internal-hexagon implants. INT J ORAL MAXILLOFAC IMPLANTS 2011;26:56-62

Biocompatibility Evaluation of Epiphany/Resilon Root Canal Filling System in Subcutaneous Tissue of Rats

Introduction: The purpose of this study was to evaluate the biocompatibility of the root canal filling system Epiphany/Resilon in connective tissue of rats. Methods: Fifteen rats were used, separated into 3 groups in accordance with its period of death (7, 21, 42 days). Four filled dentin tubes were implanted with the tested materials as follows: ERSP group, Epiphany/Resilon with Self-etch Primer; ER group, Epiphany/Resilon without primer; EG group, Endofill/gutta-percha points; and ET group, empty tube. After 7, 21, and 42 days, animals were killed, obtaining 5 samples per group. A grade from I-IV was used to graduate the inflammatory reaction. Results: Results showed that Epiphany/Resilon (ERSP and ER groups) induced a slight (II) inflammatory reaction after 42 days. However, in ER group, in which the self-etch primer was not applied, severe (IV) to moderate (III) inflammatory reactions were observed between 7 and 21 days. When compared with the EG and ET groups, it was observed that these groups presented tissue reaction ranging from slight (II, 7 and 21 days) to no inflammation (I, 42 days). Conclusions: Epiphany/Resilon root canal filling system presented satisfactory tissue reaction. It was biocompatible when tested in connective tissue of rats. (J Endod 2010;36:110-114)

Effects of the domestic use of a disclosing solution on the denture biofilm: a preliminary study

To investigate the effect of the home use of a disclosing agent on the microbial composition of denture biofilm, by means of a cross-over randomized clinical trial. Two interventions were tested during 7 days each: (i) oral and denture hygiene instructions and (ii) instructions associated with the home use of a disclosing agent (1% neutral red). Eleven participants with visible biofilm deposits over their maxillary complete dentures were randomly assigned to one of the two sequences of interventions: (i) I followed by II, and (ii) II followed by I. A washout period of 7 days was established. After each intervention, samples of denture biofilm were evaluated by DNA checkerboard hybridization for the detection of Candida spp. and 17 bacterial species. Counts were low for all the tested species, and no significant difference was found between the tested interventions ( Wilcoxon test, P > 0.05). The home use of a disclosing agent does not remarkably change the composition of denture biofilm.

Microbiology and application of the anaerobic ammonium oxidation ('anammox') process

Ten years ago, an anaerobic ammonium oxidation ('anammox') process was discovered in a denitrifying pilot plant reactor. From this system, a highly enriched microbial community was obtained, dominated by a single deep-branching planctomycete, Candidatus Brocadia anammoxidans. Phylogenetic inventories of different wastewater treatment plants with anammox activity have suggested that at least two genera in Planctomycetales can catalyse the anammox process. Electron microscopy of the ultrastructure of B. anammoxidans has shown that several membrane-bounded compartments are present inside the cytoplasm. Hydroxylamine oxidoreductase, a key anammox enzyme, is found exclusively inside one of these compartments, tentatively named the 'anammoxosome'.