Effect of gastric acid on the surface roughness and bacterial adhesion of bulk-fill composite resins

Abstract The purpose of this in vitro study was to evaluate the effect of gastric acid on the surface roughness and biofilm formation of bulk-fill composite resins. Twenty-seven samples of each composite resin were obtained: G1: Filtek Z250 XT (Z250), G2: Filtek Bulk Fill (FTK), G3: Tetric N-Ceram Bulk Fill (TTC), and G4: Aura Bulk Fill (AUR). The samples were quantitatively analyzed for surface roughness (Ra) using a roughness tester (n=15) and for biofilm formation (Cn) by the counting of colony-forming units (CFUs/mL) (n=9) in three different moments: after polishing (Ra0 and Cn0), after gastric acid immersion (Ra1 and Cn1), and after gastric acid and simulated tooth brushing (Ra2 and Cn2). Qualitative analysis through surface topography (n=3) was evaluated by scanning electron microscopy (SEM). Ra values were subjected to two-way repeated measures ANOVA, followed by Tukey's test. Cn values were subjected to Kruskal-Wallis analysis, followed by multiple comparisons analysis (α=0.05). Z250 and FTK showed significant increases in surface roughness at Ra1. There were fewer CFUs/mL on TTC and AUR in relation to those of Z250 and FTK for Cn0, Cn1 and Cn2. The SEM images showed that gastric acid increased the formation of cracks, exposure of fillers and micro cavities for all composite resins. After tooth brushing, the topographical changes were more evident but did not influence biofilm formation. The gastric acid promoted both degradation of the surfaces and bacterial adhesion for all composite resins.

Resumo O objetivo deste estudo in vitro foi avaliar o efeito do ácido gástrico na rugosidade superficial e na formação do biofilme nas resinas compostas de incremento único. Vinte e sete amostras de cada resina composta foram confeccionadas: G1: Filtek Z250 XT (Z250), G2: Filtek Bulk Fill (FTK), G3: Tetric N-Ceram Bulk Fill (TTC), and G4: Aura Bulk Fill (AUR). As amostras foram analisadas quantitativamente quanto à rugosidade da superfície (Ra) usando um rugosímetro (n=15) e para formação de biofilme (Cn) pela contagem de unidades formadoras de colônias (UFC/mL) (n=9) em três diferentes momentos: após polimento (Ra0 and Cn0), após imersão em ácido gástrico (Ra1 and Cn1), e após ácido gástrico e simulação de escovação (Ra2 and Cn2). Análise qualitative da topografia superficial (n=3) foi avaliada por meio de microscopia eletrônica de varredura (MEV). Os valores de Ra foram analisados pela ANOVA de duas vias para amostras pareadas, seguido do teste de Tukey. Os valores de Cn foram submetidos ao teste de Kruskal-Wallis, seguido da análise de comparações múltiplas (α=0,05). Z250 e FTK tiveram aumento significativo na rugosidade superficial em Ra1. Houve menos CFUs/mL para TTC e AUR em relação à Z250 e FTK em Cn0, Cn1 and Cn2. As imagens em MEV mostraram que o ácido gástrico aumentou a formação de fendas, exposição das partículas e mcrocavidades para todas as resinas compostas. Após escovação, as mudanças topográficas foram mais evidentes, mas não influenciou na formação do biofilme. O ácido gástrico promoveu degradação da superfície e adesão bacteriana para todas as resinas compostas.

In vitro evaluation of hydrolytic enzyme activity and biofilm formation of Candida parapsilosis species complex from a nosocomial environment

Abstract INTRODUCTION: Candida parapsilosis complex species, frequently found in hospital environments, have gained importance as etiological agents of candidemia. METHODS: Candida parapsilosis complex isolates from a nosocomial environment were identified and their hydrolitic enzyme activity and ability to form biofilm were characterized. RESULTS: Twenty-two C. parapsilosis sensu stricto isolates produced proteinase and three produced phospholipase. Most Candida metapsilosis isolates produced proteinase and one also produced phospholipase. All 29 isolates formed biofilms. CONCLUSIONS: The nosocomial environment may act as a reservoir for C. parapsilosis complex isolates with phenotypic features that could possibly lead to nosocomial infections and health complications in hospital patients.

Carbapenem-resistant Acinetobacter baumannii contamination in an intensive care unit

Abstract INTRODUCTION: Acinetobacter baumannii is a major pathogen causing infections in intensive care units (ICUs). In this study, we aimed to evaluate the presence of A. baumannii in an ICU environment and gloves from ICU workers and to characterize the antimicrobial resistance of the isolates in comparison with those isolated from ICU patients at the same hospital. METHODS: ICU samples were collected from March to November 2010. Isolates biochemically characterized as Acinetobacter calcoaceticus-Acinetobacter baumannii complex were evaluated by PCR targeting the 16S rDNA and bla OXA-51 genes. Antimicrobial susceptibility was determined using the disk diffusion method, and carbapenem-resistant isolates were also evaluated for the minimum inhibitory concentration of imipenem using broth microdilution. The presence of the bla OXA-23 gene was evaluated in isolates with reduced susceptibility to carbapenems. RESULTS: A. baumannii was detected in 9.5% (84) of the 886 samples collected from the ICU environment, including from furniture, medical devices, and gloves, with bed rails being the most contaminated location (23.8%; 20/84). Multidrug-resistant (MDR) A. baumannii was found in 98.8% (83/84) of non-clinical and 97.8% (45/46) of clinical isolates. Reduced susceptibility to carbapenems was detected in 83.3% (70/84) of non-clinical and 80.4% (37/46) of clinical isolates. All isolates resistant to carbapenems harbored bla OXA-23. CONCLUSIONS: We found a strong similarity between the antimicrobial susceptibility profiles of non-clinical and clinical A. baumannii isolates. Such data highlight the ICU environment as a potential origin for the persistence of MDR A. baumannii, and hence the ICU may be a source of hospital-acquired infections caused by this microorganism.

Efeito da irrigação endodôntica, com e sem ultrassom, na remoção de smear layer e bioflme / Effect of endodontic irrigation, with or without ultrasound, in removing smear layer and bioflme

Investigou-se o efeito da irrigação endodôntica, com ou sem a ativação por ultrassom, na limpeza e descontaminação dos três terços do canal radicular. Dentes foram inoculados com E. faecalis e permaneceram em cultura por 50 dias, para a formação do biofilme. Os dentes foram divididos em oito grupos de acordo com o irrigante endodôntico e o uso da ativação ultrassônica: G1 = NaOCl a 2,5% + ultrassom; G2 = clorexidina solução a 2% + ultrassom; G3 = clorexidina gel a 2% + ultrassom; G4 = H2O + ultrassom; G5 = NaOCl a 2,5%; G6 = clorexidina solução a 2%; G7 = clorexidina gel a 2% e G8 = H2O. As raízes foram clivadas e analisadas por microscopia eletrônica de varredura (MEV). As imagens foram classificadas de acordo com o nível de limpeza (presença de smear layer , 2.000x) e descontaminação (presença de bactérias, 10.000x) nos terços coronário, médio e apical. O ultrassom melhorou a habilidade de limpeza e descontaminação de todos os irrigantes endodônticos testados, principalmente do hipoclorito de sódio e da clorexidina solução. A clorexidina gel sem ultrassom teve os mais baixos valores de limpeza, contudo, seu uso combinado com o ultrassom promoveu limpeza similar ao NaOCl a 2,5%. Quanto à descontaminação, no terço apical, a clorexidina solução sem ultrassom apresentou melhores resultados do que a clorexidina gel com ultrassom. O ultrassom melhorou a habilidade de limpeza nos três terços do canal radicular, por levar o irrigante em contato com os microrganismos e debris dentinários dentro do sistema de canal, otimizando a remoção desses.

A specific polymerase chain reaction method to identify Stenotrophomonas maltophilia

Stenotrophomonas maltophilia is a multidrug-resistant nosocomial pathogen that is difficult to identify unequivocally using current methods. Accordingly, because the presence of this microorganism in a patient may directly determine the antimicrobial treatment, conventional polymerase chain reaction (PCR) and real-time PCR assays targeting 23S rRNA were developed for the specific identification of S. maltophilia. The PCR protocol showed high specificity when tested against other species of Stenotrophomonas, non-fermentative Gram-negative bacilli and 100 clinical isolates of S. maltophilia previously identified using the Vitek system.

Identification of candida spp. by phenotypic tests and PCR

The correct identification of Candida species is of great importance, as it presents prognostic and therapeutical significance, allowing an early and appropriate antifungical therapy. The purpose of this study was to identify isolates of Candida spp. from oral mucosa of 38 patients with oral candidosis evaluated in 2004 by phenotypic methods and PCR, discriminating C. albicans from the other Candida species. The tests used for phenotypic analysis were germ-tube and chlamydoconidia production, culture in CHROMAgarTM Candida, carbohydrate assimilation test, growth at 45ºC and culture in Tween 80 agar. Genotypic confirmation was performed by PCR. Phenotypic tests showed that 63.2 percent strains formed germ-tubes, 73.7 percent produced chlamydoconidia, and 63.2 percent showed green colonies in chromogenic medium, presumptively indicating C. albicans or C. dubliniensis. The carbohydrate assimilation test confirmed these results. A total of 21 percent strains were identified as C. krusei and 13.2 percent were indicative of C. tropicalis. Of these later strains, three produced chlamydoconidia. The association of other phenotypic tests with culture in Tween 80 agar identified 95.8 percent of strains as C. albicans and 4.2 percent as C. dubliniensis. All 24 strains indicative of C. albicans and C. dubliniensis were confirmed by PCR as C. albicans.

Phenotypic and genotypic characterization of Salmonella Enteritidis isolates

In order to study the epidemiology of Salmonella Enteritidis outbreaks and determine the source of contamination so that a recurrence can be avoided, detailed characterization is necessary. Thus, the purpose of this study was to verify whether rep-PCR was able to discriminate among Salmonella Enteritidis isolates. Phage typing, detection of virulence genes and antimicrobial resistance testing were also associated to rep-PCR results. One hundred and two S. Enteritidis isolates from broiler carcasses, food, human, pigs, poultry-related samples, and nine isolates from other countries were genotypically typed by REP-PCR, ERIC-PCR and BOX-PCR, collectively called rep-PCR. Phage typing, detection of virulence genes and antimicrobial resistance testing were also performed. Only three fingerprinting profiles were obtained with each rep-PCR method, with the majority of isolates belonging to the same profile. No relationship was observed between genotypic profile and year, place of isolation or source of infection. However, the less frequent rep-PCR profiles showed single antimicrobial resistance patterns. Although few strains isolated from swine were analyzed, different antimicrobial resistance patterns were observed. Furthermore, phage type 4 was not found in swine isolates. rep-PCR showed a lower discriminatory power as compared with antimicrobial resistance and phage typing, but the combination of genotypic and phenotypic methods was more discriminatory than any method alone, resulting in 48 different types.

Uma caracterização detalhada de Salmonella Enteritidis é necessária para que possa ser desenvolvido o estudo da epidemiologia dos surtos causados por este organismo, bem como a determinação da fonte de contaminação, evitando que ocorram novos surtos. Assim, o objetivo deste estudo foi verificar se a rep-PCR era capaz de diferenciar isolados de S. Enteritidis. A fagotipagem, a detecção de genes de virulência e a determinação de resistência antimicrobiana foram associadas aos resultados da rep-PCR. Cento e duas S. Enteritidis isoladas de carcaças de frango, alimentos prontos para consumo, humanos suínos, amostras relacionadas a aves, e nove isolados de outros países foram genotipicamente tipados por REP-PCR, ERIC-PCR e BOX-PCR, juntamente chamados de rep-PCR. A fagotipagem, a detecção de genes de virulência e a determinação de resistência antimicrobiana também foram realizadas. Somente três padrões de fingerprinting foram obtidos com cada método de rep-PCR, sendo que a maioria dos isolados pertenceu ao mesmo perfil. Nenhuma relação foi observada entre o perfil genotípico e o ano, o local de isolamento e a fonte de infecção. Entretanto, os perfis menos freqüentes de rep-PCR apresentaram padrões de resistência antimicrobiana únicos. Embora poucas amostras de suínos tenham sido analisadas, diferentes padrões de resistência antimicrobiana foram observados. Além disso, o fagotipo 4 não foi encontrado em isolados de suínos. A rep-PCR apresentou um menor poder discriminatório quando comparada com a resistência antimicrobiana e com a fagotipagem, mas a combinação dos métodos genotípicos e fenotípicos foi mais discriminatória do que qualquer método isolado, resultando em 48 tipos diferentes.

Detection of virulence genes in Salmonella Enteritidis isolated from different sources

A presença de três genes de virulência (invA, spvR e spvC) foi determinada em Salmonella Enteritidis isoladas de aves, suínos, humanos e alimentos. Todos os isolados foram positivos para o gene invA, 91,2% também foram positivos para o spvR e 90,2% para o spvC. Não existiu diferença significativa na prevalência dos genes de virulência entre isolados de diferentes origens. Os resultados indicaram que, provavelmente, exista um alto potencial de virulência nos isolados de S. Enteritidis caracterizados.

Phage types of Salmonella enteritidis isolated from clinical and food samples, and from broiler carcasses in Southern Brazil

272 isolates of Salmonella Enteritidis (111 isolated from frozen broiler chicken carcasses, 126 from human food and other biological materials involved in food poisoning outbreaks and 35 from different poultry materials) were selected for phage typing. From these, 111 were phage typed, 57.65 percent being classified as phage type 4, 32.43 percent as phage type 4a, 3.60 percent as phage type 6a and 0.90 percent as phage type 7, whereas 5.40 percent samples were not phage typeable. The predominance of phage type 4 is in agreement with the results published worldwide, and reinforces the need for studies related to the epidemiological meaning of these findings

Detection of virulence genes in Salmonella Enteritidis isolated from different sources

The presence of three virulence genes, invA, spvR, and spvC, was determined in Salmonella Enteritidis isolated from poultry, pigs, humans and food. All isolates were positive for the invA gene, with 91.2% being positive for spvR and 90.2% for spvC. There was no significant difference in the prevalence of the virulence genes between isolates from different sources. The results indicate that there is a putative high virulence potential for the S. Enteritidis isolates characterized.

A presença de três genes de virulência (invA, spvR e spvC) foi determinada em Salmonella Enteritidis isoladas de aves, suínos, humanos e alimentos. Todos os isolados foram positivos para o gene invA, 91,2% também foram positivos para o spvR e 90,2% para o spvC. Não existiu diferença significativa na prevalência dos genes de virulência entre isolados de diferentes origens. Os resultados indicaram que, provavelmente, exista um alto potencial de virulência nos isolados de S. Enteritidis caracterizados.

Polymerase chain reaction (PCR) for the detection of Salmonella in artificially inoculated chicken meat

The aim of this study was to develop a polymerase chain reaction (PCR) protocol for the detection of Salmonella in artificially contaminated chicken meat. Tests were performed with different dilutions of Salmonella Typhimurium or Salmonella Enteritidis cells (10-7, 10-8 or 10-9 CFU/mL) inoculated in chicken meat samples, in order to establish the limits of detection, incubation times (0, 6, 8 and 24 hours of pre-enrichment in PBW 1 percent) and three DNA extraction protocols (phenol-chloroform, thermal treatment and thermal treatment and Sephaglass). The assay was able to detect until 10-9 CFU/mL of initial dilution of Salmonella cells inoculated in chicken meat, which allows detection of Salmonella within 48 hours, including 24 hours of pre-enrichment and using the phenol-chloroform DNA extraction protocol. As the results are obtained in a shorter time period than that of microbiological culture, this procedure will be useful in the methodology for detection of Salmonella in chicken