Clinical and microbiological evaluation of the use of toothpaste containing 1% chlorhexidine and the influence of motivation on oral hygiene in patients with motor deficiency

Patients with motor deficiency have variable difficulties with mechanical plaque control, and as a consequence, the incidence of dental caries and periodontal disease can be higher in these patients. The objective of this study was to evaluate the clinical and microbiological efficacy of a toothpaste containing 1% chlorhexidine, which was used by patients with motor deficiency for 14 days. The reduction in plaque and gingival index and the impact on salivary microorganisms was evaluated. We conclude that the motivation of caregivers to carry out oral hygiene for patients with mental and motor deficiency is of great importance and is effective in reducing the formation of plaque as long as it is continuously reinforced. The use of chlorhexidine- containing toothpaste significantly reduced the plaque index and microorganism count between days 0 and 14. A reduction was also observed in the group that used a dentifrice without the chlorhexidine, but this difference was not significant. © 2010 Special Care Dentistry Association and Wiley Periodicals, Inc.

Avaliação do potencial antagônico de leveduras, visando biocontrole de deterioração por penicillium expansum

Considerable losses during apple fruit storage occur due to microbiological diseases, mainly caused by Penicillium expansum, which in addition to fruit pulp deterioration produces patulin, a mycotoxin with carcinogenic and teratogenic activity. Biological control of post-harvest disease by antagonist yeasts focused on killer toxins is an appreciable alternative to the chemical fungicides, due to the low possibility of toxic residues demonstrated during fermentative processes. Twenty out of 44 yeasts (16 isolated from fruits, 10 from corn silage and 18 from laboratory anthill), showed antagonism against spores of P. expansum. The assay in solid medium pointed the strongest nutrient competition antagonism by D. hansenii strain C1 (31 mm inhibition diameter), while D. hansenii strain C7 (15 mm) showed higher antibiosis and parasitism pattern. In the following step the extracellular activity was tested performing the assay with culture supernatant in Yeast Medium agar, where C. guilliermondii P3 was more effective against conidia germination (inhibition rate of 58.15%) while P. ohmeri showed better inhibition on micelial growth (66.17%). The antibiosis showed by both yeasts could suggest probable mechanism associated with killer phenomenon, once both strains were killer positive against sensitive reference strains (S. cerevisiae NCYC 1006 and P. kluyveri CAY-15). In order to enhance the production of antifungal substance, these yeasts were cultivated with P. expansum, but the difference between culture supernatant obtained from yeasts cultivated alone and with mould was not significant (P > 0.05). The results demonstrated that the yeasts application constitute a promising tool, enhancing the biological control of P. expansum in post-harvest diseases of apple fruit.

In vitro effect of low-level laser therapy on typical oral microbial biofilms

The aim of this study was to evaluate the effect of specific parameters of low-level laser therapy (LLLT) on biofilms formed by Streptococcus mutans, Candida albicans or an association of both species. Single and dual-species biofilms - SSB and DSB - were exposed to laser doses of 5, 10 or 20 J/cm 2 from a near infrared InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm, 0.04 W). After irradiation, the analysis of biobilm viability (MTT assay), biofilm growth (cfu/mL) and cell morphology (SEM) showed that LLLT reduced cell viability as well as the growth of biofilms. The response of S. mutans (SSB) to irradiation was similar for all laser doses and the biofilm growth was dose dependent. However, when associated with C. albicans (DSB), S. mutans was resistant to LLLT. For C. albicans, the association with S. mutans (DSB) caused a significant decrease in biofilm growth in a dose-dependent fashion. The morphology of the microorganisms in the SSB was not altered by LLLT, while the association of microbial species (DSB) promoted a reduction in the formation of C. albicans hyphae. LLLT had an inhibitory effect on the microorganisms, and this capacity can be altered according to the interactions between different microbial species.

Mineral water: A microbiological approach

The microbiological quality of bottled mineral water of various domestic brands sold in Brazil was investigated, with particular focus on the heterotrophic plate count (HPC). Neither total coliforms nor Escherichia coli were found in any 1.5 L bottle samples. Total coliforms were found in 2.9% of the small bottles, while in 20 L bottles the presence of total coliforms and E. coli was demonstrated in 15.5 and 2.4% of samples, respectively. Pseudomonas aeruginosa was detected in 4.3, 4.5 and 9.5% of small, 1.5 and 20 L bottles, respectively. In 36.4% of the samples of 1.5 L bottles, the HPC was above 500 cfu/mL. This percentage of samples with an HPC above 500 cfu/mL increased to 52.0 and 61.9% in small and 20 L bottles, respectively. Higher contamination by total coliforms, E. coli, P. aeruginosa and HPCs occurred in 20 L bottles. In conclusion, several samples in this study were outside the international quality standard for mineral water and the large number of samples with high HPCs shows that more work must be done on the use of HPC in mineral water and the damaging effects that these microorganisms may cause to humans. The bottled mineral water was confirmed as a particularly important public health problem, due to the poor microbiological quality of the products that are marketed. © IWA Publishing 2012.

Avaliação microbiológica da superfície dos implantes ortopédicos metálicos usados para fixar fraturas ósseas

In the majority of cases of bone fracture requiring surgery, orthopedic implants (screw-plate and screw) are used for osteosynthesis and the infections associated with such implants are due to the growth of microorganisms in biofilms. The objective of this study was to identify microorganisms recovered from osteosynthesis implants used to fix bone fractures, to assess the viability of the cells and the ability of staphylococci to adhere to a substrate and to determine their sensitivity/resistance to antimicrobials. After surgical removal, the metal parts of austenitic stainless steel (ASTM F138/F139 or ISO NBR 5832-1/9) were transported to the Laboratory of Clinical Microbiology, washed in buffer and subjected to ultrasonic bath at 40±2 kHz for 5 minutes. The sonicated fluid was used to seed solid culture media and cell viability was assessed under the microscope by with the aid of a fluorescent marker. The production of extracellular polysaccharide by Staphylococcus spp. was investigated by means of adhesion to a polystyrene plate. The profile of susceptibility to antimicrobials was determined by the disk diffusion assay. The most frequently isolated bacteria included coagulase-negative Staphylococcus resistant to erythromycin, clindamycin and oxacillin. Less frequent were Pseudomonas aeruginosa resistant to trimethoprim/sulfamethoxazole and ampicillin, Acinetobacter baumannii resistant to ceftazidime, Enterobacter cloacae resistant to cephalothin, cefoxitin, cefazolin, levofloxacin and ciprofloxacin, Bacillus spp. and Candida tropicalis. The observation of slides by fluorescence microscope showed clusters of living cells embedded in a transparent matrix. The test for adherence of coagulase-negative Staphylococcus to a polystyrene plate showed that these microorganisms produce extracellular polysaccharide. In conclusion, the metal parts were colonized by bacteria related to orthopedic implant infection, which were resistant to multiple antibiotics.

Connective tissue graft plus resin-modified glass ionomer restoration for the treatment of gingival recession associated with non-carious cervical lesions: Microbiological and immunological results

Objectives: It was previously reported the clinical results of placing subgingival resin-modified glass ionomer restoration for treatment of gingival recession associated with non-carious cervical lesions. The aim of this study was to evaluate the influence of this treatment on the subgingival biofilm and gingival crevicular fluid (GCF) inflammatory markers. Materials and methods: Thirty-four patients presenting the combined defect were selected. The defects were treated with either connective tissue graft plus modified glass ionomer restoration (CTG+R) or with connective tissue graft only (CTG). Evaluation included bleeding on probing and probing depth, 5 different bacteria targets in the subgingival plaque assessed at baseline, 45, and 180 days post treatments, and 9 inflammatory mediators were also assessed in the GCF. Results: The levels of each target bacterium were similar during the entire period of evaluation (p > 0. 05), both within and between groups. The highest levels among the studied species were observed for the bacterium associated with periodontal health. Additionally, the levels of all cyto/chemokines analyzed were not statistically different between groups (p > 0. 05). Conclusion: Within the limits of the present study, it can be concluded that the presence of subgingival restoration may not interfere with the subgingival microflora and with GCF inflammatory markers analyzed. Clinical relevance: This approach usually leads to the placement of a subgingival restoration. There is a lack of information about the microbiological and immunological effects of this procedure. The results suggest that this combined approach may be considered as a treatment option for the lesion included in this study. © 2012 Springer-Verlag.

Application of micronucleus test and comet assay to evaluate BTEX biodegradation

The BTEX (benzene, toluene, ethylbenzene and xylene) mixture is an environmental pollutant that has a high potential to contaminate water resources, especially groundwater. The bioremediation process by microorganisms has often been used as a tool for removing BTEX from contaminated sites. The application of biological assays is useful in evaluating the efficiency of bioremediation processes, besides identifying the toxicity of the original contaminants. It also allows identifying the effects of possible metabolites formed during the biodegradation process on test organisms. In this study, we evaluated the genotoxic and mutagenic potential of five different BTEX concentrations in rat hepatoma tissue culture (HTC) cells, using comet and micronucleus assays, before and after biodegradation. A mutagenic effect was observed for the highest concentration tested and for its respective non-biodegraded concentration. Genotoxicity was significant for all non-biodegraded concentrations and not significant for the biodegraded ones. According to our results, we can state that BTEX is mutagenic at concentrations close to its water solubility, and genotoxic even at lower concentrations, differing from some described results reported for the mixture components, when tested individually. Our results suggest a synergistic effect for the mixture and that the biodegradation process is a safe and efficient methodology to be applied at BTEX-contaminated sites. © 2012 Elsevier Ltd.

Microbiological quality of organic and conventional vegetables sold in Brazil

While searching for healthier diets, people became more attentive to organic produce. Yet, organic foods may be more susceptible to microbiological contamination because of the use of organic fertilizers, a possible source of pathogenic bacteria. In this study, 130 samples of different organic and conventional vegetable varieties sold in Brazil were analyzed for mesophilic aerobic bacteria, yeasts and molds, total coliforms, Escherichia coli and Salmonella spp. Most of the mesophilic aerobic bacteria counts in organic and conventional vegetables ranged from 6 to 7 log10 CFU/g; most of the yeasts and molds counts ranged from 5 to 6 log10 CFU/g and most of the total coliforms counts ranged from 4 to 5 log10 CFU/g. E. coli was found in 41.5% of the organic and 40.0% of the conventional vegetables, and most samples had counts ranging from 1 to 2 log10 CFU/g. Salmonella spp. was not found in any sample. Comparative analyses of the microbial counts of organic and conventional vegetables showed that some organic varieties have greater counts. However, the global results show that this is not a trend. These results indicate the need of good farming practices, and proper sanitization before consumption, to ensure food quality and safety. © 2012 Elsevier Ltd.

Assessment of the genotoxicity of two agricultural residues after processing by diplopods using the Allium cepa assay

Agroindustrial by-products and residues from treatment of sewage sludge have been recently recycled as soil amendments. This study was aimed at assessing toxic potential of biosolid, obtained from a sewage treatment plant (STP), vinasse, a by-product of the sugar cane industry, and a combination of both residues using Allium cepa assay. Bioprocessing of these samples by a terrestrial invertebrate (diplopod Rhinocricus padbergi) was also examined. Bioassay assembly followed standards of the Brazilian legislation for disposal of these residues. After adding residues, 20 diplopods were placed in each terrarium, where they remained for 30 days. Chemical analysis and the A. cepa assay were conducted before and after bioprocessing by diplopods. At the end of the bioassay, there was a decrease in arsenic and mercury. For the remaining metals, accumulation and/or bioavailability varied in all samples but suggested bioprocessing by animals. The A. cepa test revealed genotoxic effects characterized by different chromosome aberrations. Micronuclei and chromosome breaks on meristematic cells and F1 cells with micronuclei were examined to assess mutagenicity of samples. After 30 days, the genotoxic effects were significantly reduced in the soil + biosolid and soil + biosolid + vinasse groups as well as the mutagenic effects in the soil + biosolid + vinasse group. Similar to vermicomposting, bioprocessing of residues by diplopods can be a feasible alternative and used prior to application in crops to improve degraded soils and/or city dumps. Based on our findings, further studies are needed to adequately dispose of these residues in the environment. © 2013 Springer Science+Business Media Dordrecht.

Study of a series of cobalt(II) sulfonamide complexes: Synthesis, spectroscopic characterization, and microbiological evaluation against M. tuberculosis. Crystal structure of [Co(sulfamethoxazole)2(H 2O)2]·H2O

Nowadays, the research for new and better antimicrobial compounds is an important field due to the increase of immunocompromised patients, the use of invasive medical procedures and extensive surgeries, among others, that can affect the incidence of infections. Another big problem associated is the occurrence of drug-resistant microbial strains that impels a ceaseless search for new antimicrobial agents. In this context, a series of heterocyclic- sulfonamide complexes with Co(II) was synthesized and characterized with the aim of obtaining new antimicrobial compounds. The structural characterization was performed using different spectroscopic methods (UV-Vis, IR, and EPR). In spite of the fact that the general stoichiometry for all the complexes was Co(sulfonamide)2·nH2O, the coordination atoms were different depending on the coordinated sulfonamide. The crystal structure of [Co(sulfamethoxazole)2(H2O)2]·H 2O was obtained by X-ray diffraction showing that Co(II) is in a slightly tetragonal distorted octahedron where sulfamethoxazole molecules act as a head-to-tail bridges between two cobalt atoms, forming polymeric chains. Besides, the activity against Mycobacterium tuberculosis, one of the responsible for tuberculosis, and the cytotoxicity on J774A.1 macrophage cells were evaluated. © 2012 Elsevier B.V. All rights reserved.

Polymerase chain reaction assay based on ratA gene allows differentiation between Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum

Salmonella Pullorum and Salmonella Gallinarum are classified as biovars of Salmonella enterica subsp. enterica serovar Gallinarum. These salmonellae are the causative agents of Pullorum disease and fowl typhoid, respectively, and are widely distributed throughout the world. Although many developed countries have eradicated these diseases from commercial poultry, they are still the cause of significant economic loss in developing countries. When serovar Gallinarum is isolated, it is difficult to immediately differentiate between biovars because they are antigenically identical by serotyping. However, they cause distinct diseases with different epidemiology, and therefore it is important to differentiate them. This may be done biochemically but takes 2 to 3 days. In the present study, S. Pullorum and S. Gallinarum whole genomes were compared, and 1 genomic region of difference, which is part of the ratA gene, was chosen as a molecular marker for a polymerase chain reaction assay to differentiate rapidly between these organisms. In all, 26 strains of S. Gallinarum and 17 S. Pullorum strains were tested and successfully differentiated by the assay. © 2013 The Author(s).

Microplate alamarblue assay for paracoccidioides susceptibility testing

CLSI method M27-A3 is not available for use with dimorphic fungi, such as those of the Paracoccidioides genus. In this study, we developed a microdilution method and added the alamarBlue reagent to test the responses of Paracoccidioides brasiliensis and Paracoccidioides lutzii against amphotericin B and itraconazole antifungals. The test proved to be sensitive, practical, and inexpensive and can be used to monitor the activity of low-growth microorganisms and their response to various drugs. © 2013, American Society for Microbiology.

Development and validation of a D-loop mtDNA SNP assay for the screening of specimens in forensic casework

Mitochondrial DNA (mtDNA) analysis is usually a last resort in routine forensic DNA casework. However, it has become a powerful tool for the analysis of highly degraded samples or samples containing too little or no nuclear DNA, such as old bones and hair shafts. The gold standard methodology still constitutes the direct sequencing of polymerase chain reaction (PCR) products or cloned amplicons from the HVS-1 and HVS-2 (hypervariable segment) control region segments. Identifications using mtDNA are time consuming, expensive and can be very complex, depending on the amount and nature of the material being tested. The main goal of this work is to develop a less labour-intensive and less expensive screening method for mtDNA analysis, in order to aid in the exclusion of non-matching samples and as a presumptive test prior to final confirmatory DNA sequencing. We have selected 14 highly discriminatory single nucleotide polymorphisms (SNPs) based on simulations performed by Salas and Amigo (2010) [1] to be typed using SNaPShotTM (Applied Biosystems, Foster City, CA, USA). The assay was validated by typing more than 100 HVS-1/HVS-2 sequenced samples. No differences were observed between the SNP typing and DNA sequencing when results were compared, with the exception of allelic dropouts observed in a few haplotypes. Haplotype diversity simulations were performed using 172 mtDNA sequences representative of the Brazilian population and a score of 0.9794 was obtained when the 14 SNPs were used, showing that the theoretical prediction approach for the selection of highly discriminatory SNPs suggested by Salas and Amigo (2010) [1] was confirmed in the population studied. As the main goal of the work is to develop a screening assay to skip the sequencing of all samples in a particular case, a pair-wise comparison of the sequences was done using the selected SNPs. When both HVS-1/HVS-2 SNPs were used for simulations, at least two differences were observed in 93.2% of the comparisons performed. The assay was validated with casework samples. Results show that the method is straightforward and can be used for exclusionary purposes, saving time and laboratory resources. The assay confirms the theoretic prediction suggested by Salas and Amigo (2010) [1]. All forensic advantages, such as high sensitivity and power of discrimination, as also the disadvantages, such as the occurrence of allele dropouts, are discussed throughout the article. © 2013 Elsevier B.V.

Evaluation of three sampling methods for the microbiological analysis of broiler carcasses after immersion chilling

Countries have different official programs and implement different sampling methods for the detection of Salmonella on poultry carcasses. In Brazil, a 25-g sample of skin and muscle excision (SME) from the wings, neck, and pericloacal parts is used; in the European Union (EU), a 25-g sample of neck skin (NSE) is used; and, in the United States, the whole carcass is rinsed with 400 ml of diluent (WCR). In the present study, these methods were evaluated to compare Salmonella occurrence and counts of hygiene indicator microorganisms (Escherichia coli, Enterobacteriaceae, and total viable count of aerobic mesophilic bacteria) using different carcasses from the same flock and also using different analytical units taken from the same carcass. Eighty flocks, with four broiler carcasses from each, were included in this study; three broilers were sampled according to protocols from Brazil, the EU, and the United States, and the last one by all three methods. SME, NSE, and WCR provided equivalent results (P > 0.05) for Salmonella detection on broiler carcasses when using different carcasses from the same flock and when using the same carcass. The predominant serovar was Salmonella Enteritidis. For the enumeration of hygiene indicator microorganisms, WRC provided higher counts than SME or NSE (P < 0.05), when using both the same or different carcasses. Therefore, it is possible to directly compare Salmonella results in poultry carcasses when using the methods recommended by the legislative bodies of Brazil, the United States, and the EU. However, WCR provides the best results for hygiene indicator microorganisms. Copyright © International Association for Food Protection.

Estrogenic and mutagenic activities of crotalaria pallida measured by recombinant yeast assay and ames test

Background: Crotalaria pallida Ailton is a plant belonging to the Fabaceae family, popularly known as rattle or rattlesnake and used in traditional medicine to treat swelling of the joints and as a vermifuge. Previous pharmacological studies have also reported anti-inflammatory, antimicrobial and antifungal activities. Nevertheless, scientific information regarding this species is scarce, and there are no reports related to its possible estrogenic and mutagenic effects. Thus, the purpose of the present study was to investigate the estrogenic potential of C. pallida leaves by means of the Recombinant Yeast Assay (RYA), seeking an alternative for estrogen replacement therapy during menopause; and to reflect on the safe use of natural products to assess the mutagenic activity of the crude extract from C. pallida leaves, the dichloromethane fraction and stigmasterol by means of the Ames test.Methods: The recombinant yeast assay with the strain BY4741 of Saccharomyces cerevisiae, was performed with the ethanolic extract, dichloromethane fraction and stigmasterol isolated from the leaves of C. pallida. Mutagenic activity was evaluated by the Salmonella/microsome assay (Ames test), using the Salmonella typhimurium tester strains TA100, TA98, TA97 and TA102, with (+S9) and without (-S9) metabolization, by the preincubation method.Results: All samples showed estrogenic activity, mainly stigmasterol. The ethanolic extract from C. pallida leaves showed mutagenic activity in the TA98 strain (-S9), whereas dichloromethane fraction and stigmasterol were found devoid of activity.Conclusion: Considering the excellent estrogenic activity performed by stigmasterol in the RYA associated with the absence of mutagenic activity when evaluated by the Ames test, stigmasterol becomes a strong candidate to be used in hormone replacement therapy during menopause. © 2013 Boldrin et al.; licensee BioMed Central Ltd.