Abutment coating with diamond-like-carbon films to reduce implant-abutment bacterial leakage

Backgroud: The influence of diamond-like-carbon (DLC) films on bacterial leakage through the interface between abutments and dental implants of external hexagon (EH) and internal hexagon (IH) was evaluated. Film deposition was performed by PECVD (Plasma Enhanced Chemical Vapor Deposition). Sets of implants and abutments (N=180, n=30) were divided according to the connection design and the treatment of the abutment base: (1) no treatment (control); (2) DLC film deposition, and (3) Ag-DLC film deposition. Under sterile conditions, 1 µL of Enterococcus faecalis was inoculated inside the implants, and abutments were tightened. The sets were tested for immediate external contamination, suspended in test tubes containing sterile culture broth, and followed-up for five days. Turbidity of the broth indicated bacterial leakage. At the end of the period, the abutments were removed and the internal content of the implants was collected with paper points and plated in Petri dishes. They were incubated for 24 h for bacterial viability assessment and colony-forming unit (CFU) counting. Bacterial leakage was analyzed by Chi-square and Fisher exact tests (α=5%). The percentage of bacterial leakage was 16.09% for EH implants and 80.71% for IH implants (P<0.0001). The bacterial load was higher inside these implants (P=0.000). The type of implant significantly influenced the results (P=0.000), whereas the films did not (P=0.487). We concluded that: (1) IH implants showed a higher frequency of bacterial leakage and (2) the DLC and Ag-DLC films did not significantly reduce the frequency of bacterial leakage and bacteria load inside the implants.

The role of nicotine, cotinine and caffeine on the electrochemical behavior and bacterial colonization to cp-Ti

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bacterial oxycarotenoids added into broiler chickens ration modify breast meat color

Some markets around the world demand for well pigmented poultry products, what justifies the intentional use of synthetic additives in rations during breeding. The inconvenient for that practice most times is the elevated price of these products, beyond the argument of being chemicals, which may not be viable for some production systems. So, the aim of this experiment was to check on the ability of natural red oxycarotenoids produced by R. gelatinosus to modify broilers breast meat color. Bacterial biomass containing oxycarotenoids were added into Cobb broilers finisher diets at 0, 1.0, 2.0 and 3.0 g/kg and fed during 10 days (35-45 d). Five replicates (10 birds each) were performed. At days 35, 37, 39, 41, 43 and 45, 100 birds were slaughtered scalded, defeathered and eviscerated. Data on live and carcass weights were recorded. After cooling, color parameters of meat were determined on breast surface using a HunterLab XE Plus colorimeter calibrated with black and white standard tiles. Regression analysis was used for the investigation of relationships between days of consumption and biomass concentration on color values, live weight and carcass yield. Results showed that redness of breast meat was significantly influenced by the biomass concentration in a linear effect (P=0.0056) and also by the time of consumption, in a quadratic effect (P=0.0232). Days of consumption also affected yellowness and lightness with significant quadratic responses (P=0.0225 and P<0.0001, respectively). Birds live weight increased significantly during the application of the experimental diets (P<0.0001), showing no negative influence of the biomass administration. Also no deleterious effects derived from the application of the product were observed on carcass yield. So we concluded that R. gelatinosus oxycarotenoids present in the bacterial biomass can modify broilers breast meat color, performing as an alternative pigmenting additive in poultry production.

Localized periodontal disease induced by bacterial plaque and palatal radicular groove: treatment and considerations

The palatal radicular groove is a morphological tooth defect, which act as local predisposing risk factor favoring accumulation of the bacterial plaque, permitting microbial invasion via root groove way, directly into periodontal structures. A patient diagnosed with palatal radicular groove and a localized periodontal disease was treated by procedures to control bacterial action and procedures to eliminate local predisposing risk factor. To treat the periodontal bone defect a sequelae of periodontal disease, a guided tissue regeneration technique was applied by using allograft and xenograft materials associated with a resorbable demineralized bovine cortical bone membrane. The objective of surgical regenerative procedure was to recover the periodontal tissues nearly as they were before periodontal disease destruction.

Vitamin A affects haematology, growth and immune response of Nile tilapia (Oreochromis niloticus, L.), but has no protective effect against bacterial challenge or cold-induced stress

Vitamin A (vitA) is an essential nutrient that acts as an endocrine regulator of several metabolic pathways, modulating normal growth and health status of animals. Although the importance of vitA for normal haematology and immune response is well documented for higher vertebrates, there is limited information on the physiological effects of vitA on fish. Therefore, we designed a 130-day feeding trial to evaluate the effect of vitA supplementation on growth, haematology, immune function and resistance to experimental infection with Aeromonas hydrophila and cold-induced stress. A group of 320 Nile tilapia fingerlings 7.49 ± 0.19 g weight (mean ± SD) were randomly stocked into 40 250 L-aquaria and fed practical diets containing graded levels of vitA (0, 0.06, 0.12, 0.24, 0.48, 0.96, 1.92, 3.84 mg retinol (ROH) kg−1 diet. Growth, haematology, plasma protein profile and immune response were significantly affected by vitA supplementation; however, no clear protective effect of vitA supplementation on disease and cold stress resistance were observed in this study. Clinical signs of vitA deficiency were: resting and abnormal swimming behaviour, exophthalmia, haemorrhages at the base of fins and on skin, serous fluids in abdominal cavity, neutropenia, reduction in red blood cell count, haematocrit and haemoglobin evolving to high mortality rates in a short period of time. A dietary level of vitA around 1.2 mg ROH kg−1 may be required to prevent gross deficiency signs and promote proper growth and health status of Nile tilapia. VitA does not seem to have a pronounced effect on leucocyte differentiation, but clearly plays an important role on maintaining normal erythropoiesis.

Effects of seminal plasma removal on bacterial growth in stallion semen

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Control of bacterial spot of tomato and activation of enzymes related to resistance by chemicals under field conditions

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Mycobacterium bovis–Infected White-Tailed Deer (Odocoileus Virginianus): Detection of Immunoglobulin Specific to Crude Mycobacterial Antigens by ELISA

White-tailed deer (Odocoileus virginianus) have recently emerged as a source of Mycobacterium bovis infection for cattle within North America. The objective of this study was to evaluate the antibody response of M. bovis–infected deer to crude mycobacterial antigens. Deer were experimentally inoculated with M. bovis strain 1315 either by intratonsilar instillation or by exposure to M. bovis–infected (i.e., in contact) deer. To determine the time course of the response, including the effects of antigen administration for comparative cervical skin testing, serum was collected periodically and evaluated by enzyme-linked immunosorbent assay (ELISA) for immunoglobulin (i.e., IgG heavy and light chains) reactivity to mycobacterial antigens. The reactivity to M. bovis purified protein derivative (PPDb) exceeded (P < 0.05) the reactivity to M. avium PPD (PPDa) only after in vivo administration of PPDa and PPDb for comparative cervical testing of the infected deer. The mean immunoglobulin response, as measured by ELISA, of intratonsilar-inoculated deer to a proteinase K–digested whole-cell sonicate (WCS-PK) of M. bovis strain 1315 exceeded (P < 0.05) the mean of the prechallenge responses to this antigen at approximately 1 month after inoculation and throughout the remainder of the study (i.e., ~11 months). This response also exceeded (P < 0.05) that of the uninfected deer. Although this is encouraging, further studies are necessary to validate the use of the proteinase K–digested M. bovis antigens in the antibody-based assays of tuberculosis.

Bacterial diversity in rhizosphere soil from Antarctic vascular plants of Admiralty Bay, maritime Antarctica

The Antarctic is a pristine environment that contributes to the maintenance of the global climate equilibrium. The harsh conditions of this habitat are fundamental to selecting those organisms able to survive in such an extreme habitat and able to support the relatively simple ecosystems. The DNA of the microbial community associated with the rhizospheres of Deschampsia antarctica Desv (Poaceae) and Colobanthus quitensis (Kunth) BartI (Caryophyllaceae), the only two native vascular plants that are found in Antarctic ecosystems, was evaluated using a 16S rRNA multiplex 454 pyrosequencing approach. This analysis revealed similar patterns of bacterial diversity between the two plant species from different locations, arguing against the hypothesis that there would be differences between the rhizosphere communities of different plants. Furthermore, the phylum distribution presented a peculiar pattern, with a bacterial community structure different from those reported of many other soils. Firmicutes was the most abundant phylum in almost all the analyzed samples, and there were high levels of anaerobic representatives. Also, some phyla that are dominant in most temperate and tropical soils, such as Acidobacteria, were rarely found in the analyzed samples. Analyzing all the sample libraries together, the predominant genera found were Bifidobacterium (phylum Actinobacteria), Arcobacter (phylum Proteobacteria) and Faecalibacterium (phylum Firmicutes). To the best of our knowledge, this is the first major bacterial sequencing effort of this kind of soil, and it revealed more than expected diversity within these rhizospheres of both maritime Antarctica vascular plants in Admiralty Bay, King George Island, which is part of the South Shetlands archipelago. The ISME Journal (2010) 4, 989-1001; doi:10.1038/ismej.2010.35; published online 1 April 2010

Bacterial and phytoplankton dynamics in a sub-tropical estuary

Heterotrophic bacterial and biomass, production, specific growth rates and growth efficiencies were studied in July 2001 and January 2002 during both spring and neap tides, along a tidal cycle, at three sites in a subtropical estuary. Major freshwater inputs located in the Northern region led to differences in both and bacterioplankton biomass and activity along the estuary. While in the Northern region is light-limited, with mean production (PP) between 1.1 and 1.9 mu g C l(-1) h(-1) and mean specific growth rates (PSG) between 0.14 and 0.16 d(-1), the Southern region registered values as high as 24.7 mu g C l(-1) h(-1) for PP and 2.45 d(-1) (mean PP between 3.4 and 7.3 mu g C l(-1) h(-1); mean PSG between 0.28 and 0.57 d(-1)). On the other hand, maximum bacterial production (BP: 63.8 mu g C l(-1) h(-1)) and specific growth rate (BSG: 32.26 d(-1)) were observed in the Northern region (mean BP between 3.4 and 12.8 mu g C l(-1) h(-1); mean BSG between 1.98 and 6.67 day(-1)). These bacterial activity rates are among the highest recorded rates in estuarine and coastal waters, indicating that this system can be highly heterotrophic, due to high loads of allochthonous carbon (mainly derived from mangrove forest). Our results also showed that, despite that BP rates usually exceeded PP, in the Southern region BP may be partially supported (similar to 45%) by PP, since a significant regression was observed between BP and PP (r = 0.455, P < 0.001).

Spatial and Temporal Variation of Archaeal, Bacterial and Fungal Communities in Agricultural Soils

Background: Soil microbial communities are in constant change at many different temporal and spatial scales. However, the importance of these changes to the turnover of the soil microbial communities has been rarely studied simultaneously in space and time. Methodology/Principal Findings: In this study, we explored the temporal and spatial responses of soil bacterial, archaeal and fungal beta-diversities to abiotic parameters. Taking into account data from a 3-year sampling period, we analyzed the abundances and community structures of Archaea, Bacteria and Fungi along with key soil chemical parameters. We questioned how these abiotic variables influence the turnover of bacterial, archaeal and fungal communities and how they impact the long-term patterns of changes of the aforementioned soil communities. Interestingly, we found that the bacterial and fungal b-diversities are quite stable over time, whereas archaeal diversity showed significantly higher fluctuations. These fluctuations were reflected in temporal turnover caused by soil management through addition of N-fertilizers. Conclusions: Our study showed that management practices applied to agricultural soils might not significantly affect the bacterial and fungal communities, but cause slow and long-term changes in the abundance and structure of the archaeal community. Moreover, the results suggest that, to different extents, abiotic and biotic factors determine the community assembly of archaeal, bacterial and fungal communities.

Analysis of the protective potential of antigens released by Leishmania (Viannia) shawi promastigotes

Leishmania (Viannia) shawi causes cutaneous lesions in humans. Parasite antigens conferring significant protection against American tegumentar leishmaniosis (ATL) might be important for the development of effective vaccine. Therefore, this work evaluates the protective effect of antigenic fractions released by L. shawi. Antigens released by promastigotes to culture medium were concentrated and isolated by SDS-PAGE. The three main fractions LsPass1 (>75 kDa), LsPass2 (75-50 kDa) and LsPass3 (<50 kDa) were electro-eluted according with their molecular mass. Immunized BALB/c mice were challenged with L. shawi promastigotes and the course of infection monitored during 5 weeks. LsPass1-challenged mice showed no protection, however, a strong degree of protection associated to smaller lesions and high expression of IFN-gamma and TNF-alpha by CD4(+) T, CD8(+) T and double negative CD4CD8 cells was achieved in LsPass3-challenged mice. Furthermore, LsPass2-challenged mice showed an intermediated degree of protection associated to high levels of IFN-gamma, IL-4 and IL-10 mRNA. In spite of increased expression of IFN-gamma and TNF-alpha, high amounts of IL-4 and IL-10 mRNA were also detected in LsPass3-challenged mice indicating a possible contribution of these cytokines for the persistence of a residual number of parasites that may be important in inducing long-lasting immunity. Therefore, LsPass3 seems to be an interesting alternative that should be considered in the development of an effective vaccine against ATL.

Bacterial leakage along the implant-abutment interface: culture and DNA Checkerboard hybridization analyses

Objective Bacterial species have been found harboring the internal surface of dental implants as consequence of their failed connections. The aim of the present study was to compare the detection frequency of bacterial leakage from human saliva through the implantabutment interface, under non-loading conditions, using either DNA Checkerboard or culture method. Materials and methods Thirty dental implants with hexagonal platforms were connected to pre-machined abutments according to the manufacturers specifications. The assemblies were individually incubated in human saliva under anaerobic conditions for 7 similar to days at 37 degrees C. Afterward, contents from the inner parts of the implants were collected and evaluated with either DNA Checkerboard (s similar to=similar to 15) or culture (n similar to=similar to 15). Subsequently, identification and quantitation of bacterial species from saliva and implants were carried out for the group evaluated with the DNA Checkerboard method. Results Both DNA Checkerboard and culture showed positive signals of bacterial leakage in 6 of the 15 evaluated samples. Capnocytophaga gingivalis and Streptococcus mutans were the most frequently detected species harboring the internal surface of the implants followed by Veillonella parvula. Conclusion Occurrence of bacterial leakage along the implantabutment interface is comparably detected with both DNA Checkerboard hybridization and conventional culture methods.

Characterization and in vitro evaluation of bacterial cellulose membranes functionalized with osteogenic growth peptide for bone tissue engineering

The aim of this study was to characterize the physicochemical properties of bacterial cellulose (BC) membranes functionalized with osteogenic growth peptide (OGP) and its C-terminal pentapeptide OGP[10-14], and to evaluate in vitro osteoinductive potential in early osteogenesis, besides, to evaluate cytotoxic, genotoxic and/or mutagenic effects. Peptide incorporation into the BC membranes did not change the morphology of BC nanofibers and BC crystallinity pattern. The characterization was complemented by Raman scattering, swelling ratio and mechanical tests. In vitro assays demonstrated no cytotoxic, genotoxic or mutagenic effects for any of the studied BC membranes. Culture with osteogenic cells revealed no difference in cell morphology among all the membranes tested. Cell viability/proliferation, total protein content, alkaline phosphatase activity and mineralization assays indicated that BC-OGP membranes enabled the highest development of the osteoblastic phenotype in vitro. In conclusion, the negative results of cytotoxicity, genotoxicity and mutagenicity indicated that all the membranes can be employed for medical supplies, mainly in bone tissue engineering/regeneration, due to their osteoinductive properties.

INTERSPECIFIC VARIATION OF THE BACTERIAL COMMUNITY STRUCTURE IN THE PHYLLOSPHERE OF THE THREE MAJOR PLANT COMPONENTS OF MANGROVE FORESTS

Mangrove forests encompass a group of trees species that inhabit the intertidal zones, where soil is characterized by the high salinity and low availability of oxygen. The phyllosphere of these trees represent the habitat provided on the aboveground parts of plants, supporting in a global scale, a large and complex microbial community. The structure of phyllosphere communities reflects immigration, survival and growth of microbial colonizers, which is influenced by numerous environmental factors in addition to leaf physical and chemical properties. Here, a combination of culture-base methods with PCR-DGGE was applied to test whether local or plant specific factors shape the bacterial community of the phyllosphere from three plant species (Avicenia shaueriana, Laguncularia racemosa and Rhizophora mangle), found in two mangroves. The number of bacteria in the phyllosphere of these plants varied between 3.62 x 10(4) in A. schaeriana and 6.26 x 10(3) in R. mangle. The results obtained by PCR-DGGE and isolation approaches were congruent and demonstrated that each plant species harbor specific bacterial communities in their leaves surfaces. Moreover, the ordination of environmental factors (mangrove and plant species), by redundancy analysis (RDA), also indicated that the selection exerted by plant species is higher than mangrove location on bacterial communities at phyllosphere.