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Clostridioides difficile causes nosocomial diarrhoea associated with antibiotic use and immunodeficiency. Although the number of paediatric C. difficile infections (CDIs) has increased worldwide, there are few studies on the molecular characterization of strains causing CDIs among children. We report the clinical features and strain molecular characterization of a CDI in a female child with a history of liver transplantation at 7 months of age. This is the first report of the 046 ribotype causing paediatric diarrhoea.

Introduction: Dental caries is the most prevalent disease in humans and its incidence is particularly high during childhood. The use of medicinal plants is a common practice in Brazil. Objective: To evaluate the optimal antimicrobial concentration of Copaifera langsdorffii (copaiba) oil-resin, in the form of dental varnish, against Streptococcus mutans (S. mutans) in children. Methods: Twenty-four children, caries-free, aged until 6 years old, were selected to participate in this study. The varnish was applied to the occlusal surfaces of all deciduous molars. The antimicrobial activity was analyzed in saliva, whose collection was conducted in two phases: before applying the copaiba varnish and after use to verify the instantaneous effectiveness of Copaifera langsdorffii dental varnish in the reduction of S. mutans. The microbiological analysis was repeated twice, establishing dilutions of 1:10 mL and 1:100 mL. Results: Comparisons between different times within the same dilution were carried out by repeated measures analysis of variance (ANOVA) associated with Tukey’s multiple comparisons test. Comparisons of conditions prior to and after treatment were performed using the t test for paired samples and it indicated that the 1% formulation promoted a more significant decrease in the number of S. mutans colonies (p = 0,0026). Conclusion: Copaiba oil-resin, in the form of dental varnish, has antimicrobial activity against S. mutans in all the concentrations studied. Further studies to identify the long-term activity and anticaries effect of this varnish are required to establish its use in caries prevention. Keywords: Dental caries, Saliva, Streptococcus mutans

Clostridioides difficile is a Gram-positive, spore-forming, anaerobic bacillus which is the leading cause of health-care-associated infective diarrhea. The rising incidence of antibiotic resistance in pathogens such as C. difficile makes researches on alternative antibacterial products very important, especially those exploring natural products like propolis. Brazilian Red Propolis, found in the Northeast region of Brazil, is composed by products from regional plants that have the antimicrobial properties. This study aimed to evaluate the in vitro activity of Brazilian Red Propolis (BRP) against C. difficile strains in planktonic and biofilm forms. The susceptibility of four strains of C. difficile to BRP was analyzed by broth microdilution method and vancomycin was included as control drug. BRP-exposed C. difficile cells were evaluated by scanning electron microscopy (SEM). Then, the effects of BRP on growing and mature C. difficile biofilms were also evaluated. BRP minimum inhibitory concentration was 625 μg/mL against all tested strains, while vancomycin MIC range was 0.5-2 μg/mL. SEM showed the loss of homogeneity in bacterial cell wall and cell fragmentation, after BRP-exposure. BRP, at MIC, reduced (P 0.05) the biomass, matrix proteins and matrix carbohydrates of growing biofilms, and, at 8xMIC, reduced (P 0.05) the biomass and matrix proteins of mature biofilms. The present study demonstrated that BRP inhibits planktonic growth, damages cell wall, decreases biofilm growth and harms mature biofilms of C. difficile.

Clostridium difficile is a Gram-positive spore forming anaerobic bacterium and the main cause of healthcare-associated diarrhea. This study aimed to perform the phenotypic characterization and molecular typing of Clostridium difficile isolates among patients at a cancer hospital in Brazil. During 18 months, 48 diarrheic fecal samples were collected, of these 48% were positive in either one or both of the performed tests: detection of toxins A/B and culture. Clostridium difficile was recovered from four samples (17%). All strains carried toxin A and B genes, and the isolates belonged to PCR-ribotype 014/020, PGFE-type NAP4 and toxinotype XVIII. On the other hand, one isolate belonged to a novel PCR-ribotype, and PFGE-type, likewise to toxinotype IXb. The isolates showed susceptibility to metronidazole, vancomycin and moxifloxacin, and were resistant to ciprofloxacin. Finally, the findings indicate high positivity between the samples tested, suggesting an expressive importance of this infection, including detection of a novel ribotype/PFGE-type of Clostridium difficile, and show for the first time the detection of community-associated Clostridium difficile infection (CA-CDI) in these patients in Northeast Brazil. These data emphasize the importance to a better understanding of the epidemiological situation of this infection in Brazilian hospitals.

The Bacteroides fragilis group strains colonize the intestinal tract of dogs as commensal bacteria. Nevertheless, they can be opportunistic pathogens responsible for significant morbidity and mortality rates in dogs, like in oral infections, abscesses and wound infections. The purpose of this study was to evaluate antimicrobial susceptibility in B. fragilis strains isolated from dogs intestinal microbiota and to evaluate the effect of subinhibitory concentrations of some antimicrobials on biofilm formation. A total of 30 B. fragilis group strains were tested for susceptibility to ten antimicrobial agents by broth microdilution method. Thirteen B. fragilis strains were tested for biofilm formation and the biofilm producer strains were chosen to evaluate the effect of subinhibitory concentrations of six antimicrobials on biofilm formation. The isolates were susceptible to amoxicillin-clavulanic acid, metronidazole, imipenem and chloramphenicol. Tetracycline and clindamycin were active against 50% and 33% of the strains, respectively. When biofilm-forming strains were grown in the presence of sub-MICs of imipenem and metronidazole, the inhibition of biofilm formation was observed. In contrast, enrofloxacin at ½ MIC caused a significant increase in biofilm formation in two of four strains examined. In conclusion, the B. fragilis group strains isolated were susceptible to most of the antimicrobials tested and the sub-MIC concentrations of imipenem, metronidazole and clindamycin were able to inhibit the biofilm formation.

The epidemiology of Clostridium difficile infections is highly dynamic as new strains continue to emerge worldwide. Here we present a detailed analysis of a new C. difficile strain (ICC-45) recovered from a cancer patient in Brazil that died from severe diarrhea. A polyphasic approach assigned a new PCR-ribotype and PFGE macrorestriction pattern to strain ICC-45, which is toxigenic (tcdA+, tcdB+ and ctdB+) and classified as ST41 from MLST Clade 2 and toxinotype IXb. Strain ICC-45 encodes for a variant TcdB that induces a distinct CPE in agreement with its toxinotype. Unlike epidemic NAP1/027 strains, which are also classified to MLST Clade 2, strain ICC-45 is susceptible to fluoroquinolones and does not overproduce toxins TcdA and TcdB. However, supernatants from strain ICC-45 and a NAP1/027 strain produced similar expression of pro-inflammatory cytokines, epithelial damage, and oxidative stress response in the mouse ileal loop model. These results highlight inflammation and oxidative stress as common features in the pathogenesis of C. difficile Clade 2 strains. Finally, this work contributes to the description of differences in virulence among various C. difficile strains. Coordenação de aperfeiçoamento de pessoal de nivel superior y Conselho Nacional de Desenvolvimento Científico e Tecnológico/[CI-1-00830-00061.01.00/13]/FUNCAP-CAPES and PPSUS-REDEMS-CNPq-FUNCAP-SESA/Brasil Coordenação de aperfeiçoamento de pessoal de nivel superior y Conselho Nacional de Desenvolvimento Científico e Tecnológico/[12535679-0]/FUNCAP-CAPES and PPSUS-REDEMS-CNPq-FUNCAP-SESA/Brasil Ministerio de Ciencia, Tecnología y Telecomunicaciones y Consejo Nacional de Rectores/[803-B1-654]/MICITT-CONARE/Costa Rica Ministerio de Ciencia, Tecnología y Telecomunicaciones y Consejo Nacional de Rectores/[803-B4-652]/MICITT-CONARE/Costa Rica Universidad de Costa Rica/[803-B5-107]/UCR/Costa Rica Universidad de Costa Rica/[803-B5-108]/UCR/Costa Rica Consejo Nacional de Ciencia y Tecnología/[FV-0004-13]/CONICIT/Costa Rica. Fondo gestionado a través de FORINVES UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Centro de Investigación en Enfermedades Tropicales (CIET)

This study determined the species of 54 staphylococci isolates from canine otitis and their ability to produce biofilm through the Congo red agar method, confirmed by scanning electron microscopy. The most frequently identified species were S. intermedius and S. simulans. Results showed that 30% of the strains were biofilm producers.

The essential oil from Lippia gracilis Schauer (Verbenaceae) leaves was examined by GC and GC-MS. Fifteen constituents were identified. Carvacrol, p-cymene and γ-terpinene were found to be the major components. In the in vitro study, 5% solution of the Lippia gracilis Schauer oil presented antibacterial activity against Staphylococcus aureus isolated from diabetic patients with infected ulcers. The study evaluated the antibacterial activity of the 5% solution of the Lippia gracilis Schauer oil on the experimental model of diabetic adult male albino Wistar rats with leaft pelvic limb infected by Staphylococcus aureus strain. In this experiment, 28 diabetic Wistar rats were used, randomly distributed in four different groups of seven rats, (G1-white; G2-negative control; G3-positive control and G4-test). When comparing group G4 with G3, it was observed that the 5% solution presented a reduced CFU/mL level showing the antibacterial effect of the oil 24 hours after the administration of the inoculum (S .aureus without Lippia gracilis Schauer 108 ±313 versus S.aureus with Lippia gracilis Schauer 13.28±4.03). The results were expressed as mean±S.E.M. One-way analysis of the variance (ANOVA) was used. The differences between the minimum inhibitory concentration in vitro test were determined by the Tukey test (p

The aim of the present study was to identify the free amino acid content in whole saliva of children with (CE) and without early childhood caries (CF) (ECC), correlating these findings with caries experience and mutans streptococci (MS) levels in saliva.Seventy-eight healthy children, both genders, 6-71 months age, were selected to participate in the study. Following examination for dmft scores calculation, unstimulated whole saliva was collected from all 78 participants, stored at -80 degrees C, and used for amino acid analysis, on a Biochem 20 plus amino acid analyzer. Stimulated whole saliva was collected from 52 children, transported, diluted and plated on MSB agar medium for detection of MS in cfu/mL.Forty different free amino acids were identified in whole saliva, with great variation in their concentration. A statistically significant relation was found between caries experience and the presence of free proline and glycine. While proline (p=0.0182) was more frequently absent in the CF group, the absence of glycine (p=0.0397) was more often observed in the CE group. In the presence of higher levels of MS, free glycine reduced the risk of experiencing dental caries (p=0.0419). Conversely, the presence of proline was found to increase the risk of experiencing the disease (p=0.0492).The presence of free proline and absence of free glycine in children with ECC, highly contaminated with MS, increased the chances of experiencing dental caries in the present population. Further studies are needed to better understand this phenomenon.

Biofilms are assemblages of microorganisms and their associated extracellular products at an interface and typically with an abiotic or biotic surface. The study of the morphology of biofilms is important because they are associated with processes of biofouling, corrosion, catalysis, pollutant transformation, dental caries, drug resistance, and so forth. In the literature, biofilms have been examined by atomic force microscopy (AFM), which has proven to be a potent tool to study different aspects of the biofilm development on solid surfaces. In this work, we used AFM to investigate topographical changes during the development process ofEnterococcus faecalisbiofilms, which were generated on sterile cellulose nitrate membrane (CNM) filters in brain heart infusion (BHI) broth agar blood plates after 24, 36, 72, 192, and 360 h. AFM height images showed topographical changes due to biofilm development, which were used to characterize several aspects of the bacterial surface, such as the presence of extracellular polymeric substance, and the biofilm development stage. Changes in the development stage of the biofilm were shown to correlate with changes in the surface roughness as quantified through the mean roughness.

Intestinal mucositis is a common side effect of irinotecan-based anticancer regimens. Mucositis causes cell damage, bacterial/endotoxin translocation and production of cytokines including IL–1 and IL–18. These molecules and toll-like receptors (TLRs) activate a common signaling pathway that involves the Myeloid Differentiation adaptor protein, MyD88, whose role in intestinal mucositis is unknown. Then, we evaluated the involvement of TLRs and MyD88 in the pathogenesis of irinotecan-induced intestinal mucositis. MyD88-, TLR2- or TLR9-knockout mice and C57BL/6 (WT) mice were given either saline or irinotecan (75 mg/kg, i.p. for 4 days). On day 7, animal survival, diarrhea and bacteremia were assessed, and following euthanasia, samples of the ileum were obtained for morphometric analysis, myeloperoxidase (MPO) assay and measurement of pro-inflammatory markers. Irinotecan reduced the animal survival (50%) and induced a pronounced diarrhea, increased bacteremia, neutrophil accumulation in the intestinal tissue, intestinal damage and more than twofold increased expression of MyD88 (200%), TLR9 (400%), TRAF6 (236%), IL–1β (405%), IL–18 (365%), COX–2 (2,777%) and NF-κB (245%) in the WT animals when compared with saline-injected group (P

Comparou-se o perfil de isolamento microbiano de amostras coletadas de cães com otite média (OM) e externa (OE) associadas. Sessenta e quatro cães com otite média e externa foram avaliados durante 10 meses. Amostras dos condutos auditivos externos foram coletadas com auxílio de swabs estéreis e aquelas do ouvido médio pela técnica cirúrgica da osteotomia da bula timpânica. Os microrganismos foram cultivados e identificados de acordo com os métodos previamente descritos, e a susceptibilidade a antimicrobianos avaliada pelo método de difusão em ágar estabelecido pelo National Committee for Clinical Laboratory Standards. Bactérias anaeróbias estritas não foram isoladas em qualquer amostra. Todas as amostras de OE mostraram crescimento bacteriano e/ou fúngico. Na OM, a positividade nas culturas foi de 48%. Os microrganismos mais isolados na OE foram: Bacillus sp. (26,9%), M. pachydermatis (22,2%) e S. intermedius (21,7%); na OM foram: S. intermedius (32,5%), S. aureus subsp. aureus (22,5%) e bacilos Gram-negativos não fermentadores (10,0%). Observou-se alguma diferença no perfil de isolamento entre os quadros de OM e OE em 96,7% dos animais. Verificou-se elevada resistência de cepas de S. intermedius à penicilina G, ampicilina, eritromicina, tetraciclina e clindamicina. The isolation pattern of samples collected from dogs with both media (OM) and extern otitis (OE) was compared. Sixty-four dogs suffering from those conditions were studied over a 10-month period. Samples from the external ear were collected with sterile swabs while those from the middle ear were collected by osteotomy of the tympanic bulla. The microorganisms were cultured and identified according to methods previously described and to the susceptibility of antimicrobials according to the National Committee for Clinical Laboratory Standards agar diffusion test. Anaerobic bacteria were not isolated in any sample. All samples from OE showed bacterial and/or fungal growth. Concerning media otitis, this percentage was 48%. The most common microorganisms isolated from OE were Bacillus sp. (26.9%), M. pachydermatis (22.2%) and S. intermedius (21.7%) and those from OM were S. intermedius (32.5%), S. aureus subsp. aureus (22.5%) and non-fermentative Gram-negative rods (10.0%). Some difference was observed in the isolation pattern between media and extern otitis in 96.7% of the animals. High resistance rates of S. intermedius strains to penicillin G, ampicillin, erythromycin, tetracycline and clindamycin were found.

This study evaluated the middle ear microbiota and antimicrobial susceptibility patterns from strains isolated from dogs with otitis media. A total of 62 dogs obtained from Zoonoses Control Center of Fortaleza City Ceará State / Brazil were studied over a 10-month period (August/2003 to June/2004). Of the total, 46.8% (n=30) of the animals were positive for otitis media and the infection was monomicrobial in 76.6% of them. The most frequent isolated agents were coagulase-positive Staphylococci (CPS-55%) and Pseudomonas sp (10%). For S. intermedius (n=13) and S. aureus subsp aureus (n=9), respectively, the greater resistance rates were observed using penicillin G (30.76% and 44.44%), ampicillin (7.69% and 44.44%), erythromycin (23.07% and 44.44%), clindamycin (23.07% and 44.44%) and thrimethropim/sulfamethoxazol (15.38% and 33.33%). O presente trabalho avaliou a microbiota do ouvido médio e os perfis de susceptibilidade a antimicrobianos de cepas isoladas de cães com otite média. Um total de 62 cães obtidos no Centro de Controle de Zoonoses de Fortaleza Ceará / Brasil foram estudados no período de 10 meses (agosto/2003 a junho/2004). Do total de animais, 46.8% (n=30) foram positivos para otite media e a infecção envolvida era monomicrobiana em 76,6% dos casos. Os agentes isolados com maior frequência foram Staphylococcus coagulase-positiva (CPS-55%) e Pseudomonas sp (10%). Considerando-se S. intermedius (n=13) e S. aureus subsp aureus (n=9), respectivamente, as mais elevadas taxas resistência foram observadas frente a: penicilina G (30,76% e 44,44%), ampicilina (7,69% e 44,44%), eritromicina (23,07% e 44,44%), clindamicina (23,07% e 44,44%) e trimetropim/sulfametoxazol (15,38% e 33,33%).

Diabetes mellitus (DM) é uma doença progressiva que apresenta complicações freqüentes, em especial a infecção nos pés. Realizamos um estudo prospectivo com 141 pacientes com DM e úlceras infectadas nos pés, conduzido no Centro Integrado de Diabetes e Hipertensão da UFC no período de março/2000 a novembro/2001. A maioria apresentava infecção avaliada como graus I e II da classificação de Wagner. As amostras foram cultivadas utilizando-se meios seletivos, e a identificação bacteriana e os antibiogramas foram realizados através de metodologia convencional e automatizados. Os patógenos mais freqüentemente isolados foram as enterobactérias (83,7%), Staphylococcus aureus (43,3%) e bactérias anaeróbias (17%). Cepas de Streptococcus pyogenes foram isoladas de 7,8% dos pacientes. Cepas produtoras de beta-lactamase de espectro ampliado e cepas de Staphylococcus aureus resistentes à oxacilina foram isoladas de 6% e 11,6% dos pacientes, respectivamente. A resistência aos antimicrobianos vem se tornando cada vez mais comum, mesmo em pacientes com infecção adquirida na comunidade. Investimentos em diagnóstico etiológico das infecções e estratégias racionais no uso de antimicrobianos fazem-se necessários na tentativa de evitar tratamentos inadequados e suas já conhecidas conseqüências.

Summary Clostridium difficile is the most important cause of nosocomial diarrhea, mainly associated with antibiotic use and immunodeficiency. Although, an increased incidence of community-acquired C. difficile infection (CA-CDI) has been reported worldwide, this infection has been under-diagnosed in Latin America. This is the first report of a CA-CDI case in Latin America, in an HIV-positive patient with cancer.

Background: WHO guidelines recommend zinc supplementation as a key adjunct therapy for childhood diarrhea in developing countries, however zinc’s anti-diarrheal effects remain only partially understood. Recently, it has been recognized that low-grade inflammation may influence stunting. In this study, we examined whether oral zinc supplementation could improve weight, intestinal inflammation, and diarrhea in undernourished weanling rats. Methods: Rats were undernourished using a northeastern Brazil regional diet (RBD) for two weeks, followed by oral gavage with a saturated lactose solution (30 g/kg) in the last 7 days to induce osmotic diarrhea. Animals were checked for diarrhea daily after lactose intake. Blood was drawn in order to measure serum zinc levels by atomic absorption spectroscopy. Rats were euthanized to harvest jejunal tissue for histology and cytokine profiles by ELISA. In a subset of animals, spleen samples were harvested under aseptic conditions to quantify bacterial translocation. Results: Oral zinc supplementation increased serum zinc levels following lactose-induced osmotic diarrhea. In undernourished rats, zinc improved weight gain following osmotic diarrhea and significantly reduced diarrheal scores by the third day of lactose intake (p < 0.05), with improved jejunum histology (p < 0.0001). Zinc supplementation diminished bacterial translocation only in lactose-challenged undernourished rats (p = 0.03) compared with the untreated challenged controls and reduced intestinal IL-1β and TNF-α cytokines to control levels. Conclusion: Altogether our findings provide novel mechanisms of zinc action in the setting of diarrhea and undernutrition and support the use of zinc to prevent the vicious cycle of malnutrition and diarrhea.

Bacteroides fragilis colonizes dog guts both as a commensal and as an opportunistic pathogen. This study aims to evaluate virulence factors of 13 B. fragilis strains isolated from dog intestinal tracts and their ability for biofilm formation. Capsules were detected in all the evaluated strains. A total of 61.5% of all strains were biofilm producers. These attributes most likely play an important role in B. fragilis persistent colonization in the gut.

A total of 30 strains of Fusobacterium sp. isolated from gengival sulcus of 28 equines was studied in the period between January/1997 and April/1998. Bacterial isolation and antimicrobial resistance were determined by biochemical analyses and by the disk elution method. The species most frequently isolated was Fusobacterium nucleatum. The following resistance rates were observed: 10% for penicillin G, 87% for eritromycin, and 97% for sulfonamide. Theses results emphasize the need for monitoring the susceptibility patterns of this important bacterial group frequently isolated from normal sites in equines.

Clindamycin resistance was trasferred by a conjugation-like process from Bacteroides thetaiotaomicron 52, a multiple antibiotic-resistant strain isolated from clinical specimens, to other Bacteroides species. A possible association between a plasmid detected in the donor strain and clindamycin resistance is discussed.Resistência à clindamicina foi transferida através de processo semelhante à conjugação, de uma cepa multirresistente e isolada de espécime clínico, B. thetaiotaomicron 52, para outra espécie de Bacteroides. Uma possível associação entre um plasmídio detectado na cepa doadora e a resistência à clindamicina é discutida.

O tecido perirradicular de 30 casos cirúrgicos foi submetido a exame microbiológico e histopatológico. Foram isoladas 137 cepas bacterianas das 30 lesões periapicais estudadas. Do total de bactérias isoladas, 90 (65,7%) foram caracterizadas como anaeróbias estritas, 40 (29,2%) como anaeróbias facultativas e 7 (5,1%) como aeróbias estritas. Fusobacterium nucleatum, bacilos Gram-negativos pigmentados anaeróbios estritos, Peptostreptococcus sp, Streptococcus mitis e bacilos Gram-positivos não esporulados anaeróbios estritos foram, em ordem decrescente de freqüência, as bactérias mais comumente isoladas das lesões periapicais. A análise histopatológica demonstrou prevalência de granuloma periapical. O estudo de sensibilidade a antimicrobianos foi realizado em 16 cepas de Fusobacterium nucleatum. Os antimicrobianos utilizados foram penicilina, cefoxitina, eritromicina, metronidazol e tetraciclina. Todas as amostras apresentaram suscetibilidade à cefoxitina, tetraciclina e metronidazol; os percentuais de resistência à penicilina e à eritromicina foram 12,5 e 68,8, respectivamente. Os percentuais de resistência de Fusobacterium nucleatum, uma espécie bacteriana importante na patogênese das lesões periapicais, à penicilina e à eritromicina, enfatizam a necessidade de outros estudos de resistência a antimicrobianos de bactérias isoladas em infecções odontogênicas.Periapical tissue from 30 individuals requiring dental surgery was examined histologically and cultured for the presence of bacteria. A total of 137 strains of bacteria were isolated and of the isolates, 65.7% were strict anaerobes, 29.2%, facultative anaerobes and 5.1%, strict aerobes. Fusobacterium nucleatum was the species most frequently isolated, followed by pigmented strict anaerobes, Peptostreptococcus sp and Streptococcus mitis. In all cases more than one bacterial species was found. The histological analysis demonstrated prevalence of periapical granuloma. Fusobacterium nucleatum isolated from 16 patients was tested for susceptibility to penicillin, cefoxitin, erythromycin, metronidazole and tetracycline. All strains were susceptible to cefoxitin, metronidazole and tetracycline and the resistance rates to penicillin and erythromycin were 12.5 and 68.8% respectively. The resistance rates of F. nucleatum, an important bacterium in periapical infection to penicillin and erythromycin emphasize the need to further study antimicrobial resistance of bacteria isolated from odontogenic infections.

Essential oils of many plants have been previously tested in the treatment of oral diseases and other infections. This study was a randomized, double-blind, in parallel with an active control study, which aimed to evaluate the efficacy of three formulations of the Lippia sidoides Cham. essential oil (LSO) in the reduction of salivary Streptococcus mutans in children with caries. 81 volunteers, aged 6-12 years, both genders, with caries, were recruited to participate in this study, and randomly assigned to either one of five different groups. Each group received topical treatment with either 1.4% LSO toothpaste, 1.4% LSO gel, 0.8% LSO mouthwash, 1% chlorhexidine gel, or 0.12% chlorhexidine mouthwash. A 5-ml volume of each gel was placed inside disposable trays, and applied for 1 min, every 24h, for 5 consecutive days. The mouthwash groups used 5-ml volume of a mouthwash inside disposable syringes. In the toothpaste group, children brushed their teeth for 1 min, once a day for 5 days. Saliva was collected before and after treatment. MS colonies were counted, isolated and confirmed through biochemical tests. Differences in MS levels measured in different days within the same treatment group was only verified with LSO toothpaste, chlorhexidine gel and chlorhexidine mouthwash. Comparison between groups of LSO mouthwash, toothpaste and gel showed that the toothpaste group expressed significantly lower MS levels than the mouthwash and gel groups at day-30. Chlorhexidine significantly reduced MS levels after 5 days of treatment, but these levels returned to baseline in other periods of the study. LSO toothpaste reduced MS levels after 5 days of treatment, and MS levels remained low and did not return to baseline during subsequent analysis. Hence, LSO toothpaste demonstrated the most long-lasting MS reduction in saliva, whereas other LSO formulations did not effectively reduce MS levels in children with dental caries.

The present work aimed to evaluate the effectiveness ofPlectranthus amboinicus(Lour.) Spreng against MRSA clinical isolates. The in vitro antimicrobial activity of the hydroalcoholic extract (HE), the ethyl acetate (EA) fraction and its subfractions were determined by broth microdilution and bioautography against MRSA clinical isolates. The microdilution checkerboard method was used to assess in vitro drug combination studies. To induce abscess formation, bacterial suspensions were added to Citodex and inoculated subcutaneously into male Swiss mice. The treatment protocol consisted of 2 doses of HE, the EA fraction or vancomycin introduced intraperitoneally into mice 3 and 12 h after infection. The EA fraction and its subfractions presented the lowest minimal inhibitory concentrations (MIC, 0.25 to 0.5 mg/mL). The plant samples were bacteriostatic at 2x and 4x MIC and bactericidal at 100 mg/mL. The EA fraction presented synergism with vancomycin and an additive effect with ciprofloxacin. A significant reduction of abscess volume, bacterial cell counts in abscess slurries, and inflammatory scores was observed in the HE and EA fraction-treated groups. The samples were effective in treating the animals in a dose-dependent fashion. The present study proved the effectiveness ofP. amboinicusfractions against MRSA using in vitro and in vivo assays.

Background As dietary management during early childhood is a great barrier in caries control, there is a need for the identification of intrinsic risk factors, capable of allowing the use of a more cost-effective approach to early childhood caries (ECC). Objective To evaluate the salivary peptide profile of children with and without ECC and its association with caries experience. Methods One hundred and six 10- to 71-month-old children participated in the study. Caries experience was determined through the visual/tactile method, based on the number of decayed, missing, and filled teeth, and surface scores (dmft/dmfs). Whole saliva was collected for mutans streptococci (MS) detection and peptide analysis. Results Chromatograms from CF (children without caries experience, n = 58) and CE (children with caries experience, n = 48) saliva pools expressed different patterns. Identification of molecular masses suggested the presence of nine peptides. Three of them were significantly related with caries experience. HNP-3 (α-defensin 3) (P = 0.019) and HBD-3 (β-defensin 3) (P = 0.034) reduced the chances of experiencing ECC. Proline-rich peptides IB-4 significantly increased caries experience (P = 0.035). Age (P = 0.020) and MS counts (P = 0.036) increased caries experience; however, gender was not associated with dental caries (P = 0.877). Conclusion Specific salivary peptides of CF or CE children in early childhood predispose to a higher or lower risk of caries experience.

This study's purposes were to: evaluate the antibacterial effect of chemomechanical instrumentation and a calcium hydroxide-based dressing in primary teeth with pulp necrosis secondary to trauma; and detect the presence of Fusobacterium nucleatum and black-pigmented rods in the canals of these teeth.Microbiological sample collections (MSCs) were obtained: after coronal access to the canals of primary incisors (in 18 teeth; MSC 1); after chemomechanical instrumentation (in 10 teeth; MSC 2); and 72 hours after removal of intracanal medication (in 18 teeth; MSC 3). These samples were sent for microbiological processing. The results were analyzed statistically via chi-square, analysis of variance, and Games-Howell tests (P.05).Micro-organisms were isolated in approximately 94%, 10%, and 83% of canals, respectively, in MSCs 1, 2, and 3. There was a statistically significant difference only between MSCs 1 and 2 (P.03). F nucleatum and black-pigmented rods were detected in approximately 56% and 11% of canals in MSC 1, respectively.Chemomechanical instrumentation and calcium hydroxide-based dressing have an anti-bacterial effect by significantly reducing the number of micro-organisms in the main root canal. They showed a limited efficacy, however, and did not prevent bacterial regrowth after endodontic therapy in primary teeth with pulp necrosis secondary to trauma.

The essential oil from the leaves of Myrcia ovata Cambess., commonly used in Brazil for the treatment of gastric illnesses, was screened for antimicrobial activity and action in the formation of microbial biofilms by Enterococcus faecalis. The oil was obtained by hydrodistillation using a clevenger-type system. Its chemical composition was analyzed using GC and GC-MS. Both MIC and MBC of the essential oil were determined by broth microdilution techniques and agar dilution method. The essential oil showed antimicrobial activity against E. faecalis, Escherichia coli, Pseudomonas aeruginosa, Salmonella choleraesuis, Staphylococcus aureus, Streptococcus pneumoniae and Candida parapsilosis. The results showed that the essential oil of M. ovata Cambess. was effective against the formation of biofilm by E. faecalis when compared with the control. Four volatile compounds, representing 92.1 % of the oil, were identified and geranial was the major component (50.4 %). At the best of our knowledge, this is the first report of the chemical composition and antimicrobial activity of the essential oil from leaves of M. ovata.

Research has demonstrated the need for identifying a novel antimicrobial agent for topical use in the pediatric dental population. The essential oil of Lippia sidoides Cham. (LSO) has been described as having favorable biological properties, and a broad in vitro and in vivo antimicrobial spectrum against bacteria and yeast infections. Our aim was to determine a dose and formulation of LSO, acceptable for clinical testing in a pediatric population with dental caries. Thirty-seven 6–12-year old children were selected to participate in this study, and randomly allocated to receive different concentrations of either a gel (0.8%, 1%, 1.2% and 1.4%) or a mouth rinse (0.6%, 0.8%, 1% and 1.2%) formulation. The highest percentage MS reduction was observed with 0.8% mouth rinse and 1.4% gel. The efficacy of these concentrations was compared with a Thy–Car mixture formulated as a mouth rinse and gel treatments in 11 children. Saliva was collected after a single application of the antimicrobial treatment to establish effectiveness against MS. Both rinse (p

Background: Bisphosphonates are potent inhibitors of bone resorption and were shown to inhibit bone resorption in experi- mental periodontitis by unknown mechanisms. We studied the effect of the aminobisphosphonate sodium alendronate (SA) in experimental periodontitis. Wistar rats were subjected to liga- ture placement around the second upper left molars. Methods: Animals were treated with SA 0.01 to 0.25 mg/kg subcutaneously (sc), either 1 hour before (prophylactic) or starting 5 days after (therapeutic) periodontitis induction and daily until the rats were sacrificed (11 days). Controls received saline. Animals were weighed daily. Alveolar bone loss was measured as the difference (in millimeters) between the cusp tip and the alveolar bone. The periodontium and the sur- rounding gingivae were examined at histopathology, and the neutrophil influx into the gingivae was assayed using myeloper- oxidase activity. The local bacterial flora was assessed through culture of the gingival tissue in standard aerobic and anaerobic media. Results: Alveolar bone loss was significantly and dose de- pendently inhibited by SA either as a prophylactic or therapeu- tic treatment compared to the control. SA reduced tissue lesion at histopathology, with partial preservation of the periodonti- um, coupled to decreased myeloperoxidase activity compared to the control. The reduced neutrophil influx was also shown in carrageenan-induced peritonitis, used as a control experiment for this parameter. SA also significantly inhibited the growth of pigmented bacilli and Fusobacterium nucleatum, which are im- portant in the pathogenesis of periodontal disease. SA also inhibited the in vitro growth of isolated Peptostreptococcus sp. Conclusion: Sodium alendronate preserves alveolar bone re- sorption and has anti-inflammatory and antibacterial activities inexperimentalperiodontitis. J Periodontol 2005;76:1901-1909

The occurrence of canine externa otitis in Fortaleza-Ceará is reported. About 91.5% of the animals with clinical signs were positive to bacterial culture. Among all infections, 49.5% were mixed infections and the most common pathogens were Staphylococcus spp coagulase negative or positive and Pseudomonas aeruginosa. The most effective antimicrobials for Staphylococcus coagulase negative were: the quinolones, the aminoglicoside netilmycin and the beta-lactams, excepted ampicillin, penicillin and oxacilin; for Staphylococcus coagulase positive were: cefotoxin, amoxicillin + clavulanic acid, imipenem, netilmycin and cephatoxin; for Pseudomonas aeruginosa were: ciprofloxacin, tobramycin and imipenem.

We bacteriologically analyzed 156 species of Enterobacteriaceae, isolated from 138 patients with community-acquired diabetic foot ulcers, in a prospective study made at a diabetic center and at the Federal University of Ceará, Brazil, from March, 2000, to November, 2001. The samples were cultured using selective media, and identification, susceptibility tests and detection of plasmid-mediated-extended-spectrum-beta-lactamase (ESBL) producing strains were made with conventional and automated methods. The most frequently occurring pathogens were K. pneumoniae (21.2%), Morganella morganii (19.9%) and E. coli (15.4%). High resistance rates were noted for ampicillin, first generation cephalosporin, trimethoprim/sulfamethoxazole, tetracycline, amoxicillin-clavulanic acid and chloramphenicol. ESBL-producing strains were detected in 6% of the patients. Resistance among gram-negative bacteria has become increasingly common, even in community-acquired infections.

Objetivo: determinar a presença de infecção assintomática do líquido amniótico em gestantes, identificar os agentes bacterianos envolvidos na infecção e determinar o perfil de suscetibilidade a antimicrobianos in vitro. Métodos: foram obtidas 81 amostras de líquido amniótico, colhidas por amniocentese, em gestantes sem sinais de trabalho de parto e sem suspeita clínica de infecção, atendidas na Maternidade Escola Assis Chateaubriand, entre agosto/97 e janeiro/99. Pesquisou-se a presença de bactérias aeróbias, anaeróbias estritas/facultativas e micoplasmas genitais. As bactérias anaeróbias foram identificadas pelo sistema ATBÒ (Bio-Mérieux) e os micoplasmas pelo kit Micoplasmas ISTÒ (Bio-Mérieux). Resultados: entre as amostras obtidas, oito (9,8%) apresentaram culturas positivas, sendo que em duas foram identificadas duas espécies bacterianas. Os patógenos isolados foram: Ureaplasma urealyticum (7 casos, 8,6%), Mycoplasma hominis (1 caso, 1,2%) e Peptostreptococcus sp (2 casos 2,4%). O padrão de resistência aos antimicrobianos caracterizou-se pela maior resistência dos micoplasmas à eritromicina (37,5%) e nenhuma resistência às ciclinas. Conclusões: o percentual de infecções assintomáticas foi muito elevado, havendo necessidade de serem realizadas novas pesquisas para avaliar as conseqüências da infecção subclínica nas grávidas e em seus conceptos, que envolvam métodos que identifiquem micoplasmas genitais, já que foram as bactérias mais freqüentemente isoladas. Purpose: to determine the presence of asymptomatic amniotic fluid infection in pregnant women, to identify the bacterial agents involved in the infection and to determine the antimicrobial susceptibility in vitro. Methods: amniotic fluid samples were obtained by amniocentesis from 81 pregnant women without labor signs and without suspucion of clinical infection, attended at Maternidade Escola Assis Chateaubriand from August 1997 to January 1999. The presence of aerobic bacteria, strict/facultative anaerobic bacteria and genital mycoplasmas was investigated. The anaerobic bacteria were identified by the ATB SystemÒ (Biolab Mérieux) and mycoplasmas by the IST MycoplasmaÒ kit (Biolab-Mérieux). Results: among the obtained samples, eight (9.8%) showed positive culture and in two samples two different strains were identified. The isolated pathogens were Ureaplasma urealyticum (7 cases, 8.6%), Mycoplasma hominis (1 case, 1.2%) and Peptostreptococcus sp (2 cases, 2.4%). The antimicrobial susceptibility was characterized by great mycoplasma resistance to erythromycin (37.5%) and no resistance to cyclins. Conclusions: the percentage of asymptomatic infections was high, and furthe research is necessary to evaluate the asymptomatic infection consequences in pregnant women and their newborns, involving methods that identify genital mycoplasmas, which were the most frequently isolated bacteria.

Um total de 40 cepas do grupo B. fragilis foi isolada de espécimes clínicos em dois centros hospitalares de Fortaleza no período de 1993 a 1997. A espécie mais frequentemente isolada foi Bacteroides fragilis (19 cepas) tendo a maioria dos microrganismos sido isolada de infecção intra-abdominal e ferida cirúrgica infectada. Foi traçado o perfil de sensibilidade à cefoxitina, cefoperazona e associação ticarcilina-ácido clavulânico, utilizando-se o método de referência de diluição em ágar. Todas as espécies testadas apresentaram sensibilidade à ticarcilina-ácido clavulânico (128/2mig/ml). Percentuais de resistência de 15 e 70% foram detectados para cefoxitina (64mig/ml) e cefoperazona (64mig/ml) respectivamente. A espécie B. fragilis apresentou os menores percentuais de resistência quando comparada com as demais espécies do grupo. Estes resultados regionais permitem uma melhor orientação na escolha deste grupo de antibióticos, para profilaxia ou terapêutica, principalmente com relação à cefoxitina que é frequentemente empregada nos centros hospitalares estudados. A total of 40 strains of the B. fragilis group was isolated from clinical specimens in two hospital centers in Fortaleza from 1993 to 1997. The most frequently isolated species was Bacteroides fragilis (19 strains) and most isolates came from intra-abdominal and wound infections. The susceptibility profile was traced for cefoxitin, cefoperazone and ticarcillin-clavulanate by using the agar dilution reference method. All isolates were susceptible to ticarcillin-clavulanate (128/2mug/ml). Resistance rates of 15 and 70% were detected to cefoxitin (64mug/ml) and cefoperazone (64mug/ml), respectively. Such regional results permit a better orientation in choosing this group of antibiotics for prophylaxis and therapy especially in relation to cefoxitin, which is frequently used in the hospital centers studied.

A total of 83 clinical isolates of the Bacteroides fragilis group collected from two Brazilian hospitals during 1993 and 1994, were tested for susceptibility to six antimicrobial agents by using an agar dilution method. The species most frequently isolated was B. fragilis (63.8%), followed by B. thetaiotaomicron (15.6%), B. vulgatus (9.6%) and B. distasonis (7.2%). The various species of the B. fragilis group had different patterns of resistance; B. fragilis was the most susceptible species in both medical centers. All isolates were susceptible to metronidazole and chloramphenicol. The following resistance rates were observed: 89.1% (Penicillin G); 77.1% (tetracycline); 18% (clindamycin) and 7.2% (cefoxitin). These results reflect the value of identification of clinical important isolates and the periodic determination of B. fragilis group susceptibility patterns at each medical institution.

Epidemiological aspects and the antimicrobial susceptibility profile of the Bacteroides fragilis group isolated from clinical and human intestinal specimens were examined in this study. B. fragilis group strains were isolated from 46 (37%) of 124 clinical specimens and the source of the samples was: Blood culture (3), intraabdominal infection (27), brain abscess (2), soft tissue infection (17), respiratory sinus (3), pleural aspirate (9), breast abscess (3), surgical infected wound (22), pelvic inflammatory disease (22), chronic otitis media (9) and miscellaneous (7). Intraabdominal and soft tissue infections were responsible for more than half of the clinical isolates. Susceptibility to penicillin, cefoxitin, tetracycline, metronidazole, chloramphenicol and clindamycin was examined. All isolates were susceptible to metronidazole and chloramphenicol. For clindamycin and cefoxitin the resistance rates observed were 21.7% and 10.9% respectively. Susceptibility profiles varied among the different species tested. A total of 37 species of B. fragilis group isolated from intestinal microbiota of individuals who had no antimicrobial therapy for at least 1 month before the sampling was also examined. All strains were also susceptible to chloramphenicol and motronidazole and the resistance rates to clindamycin and cefoxitin were 19.4% and 5.4% respectively. A few institutions, in Brazil, have monitored the antimicrobial susceptibility of B. fragilis group strains isolated from anaerobic infections. The resistance rates to cefoxitin and clindamycin and the variation in susceptibility patterns among the species isolated in this study emphasize the need for monitoring of susceptibility patterns of B. fragilis group organisms isolated, especially at our University Hospitals. Alguns aspectos epidemiológicos e o perfil de sensibilidade a antimicrobianos de amostras do grupo B. fragilis isoladas de espécime clínico e microbiota intestinal humana foram delineados neste estudo. As espécies do grupo B. fragilis foram isoladas de 46 (37%) de 124 espécimes clínicos, como segue: hemocultura (3) infecção intra-abdominal (27), abscesso cerebral (2), infecção de tecido mole (17), seios da face (3), aspirado pleural (9), abscesso pulmonar (3), ferida cirúrgica (22), doença inflamatória pélvica (22), otite média crônica (9) e diversos (7). Mais da metade destes microorganismos foram isolados de infecção intra-abdominal e infecção de tecido mole. Os antimicrobianos testados foram Penicilina G, Cefoxitina, Cloranfenicol, Metronidazol, Tetraciclina e Clindamicina. Todas as amostras estudadas apresentaram-se sensíveis ao Cloranfenicol e ao Metronidazol. Resistência à clindamicina e cefoxitina foi observada em 21.7 e 10.9% dos microrganismos para cada droga respectivamente. Variação na sensibilidade aos antimicrobianos entre as espécies do grupo foi detectada. O perfil de sensibilidade a antimicrobianos de amostras do grupo B. fragilis isoladas da microbiota intestinal de 37 indivíduos que não faziam uso de antimicrobianos nos últimos 30 dias antes do exame foi também estudado. Todas as amostras apresentaram-se sensíveis ao cloranfenicol e metronidazol sendo de 19.4% e 5.4% os percentuais de resistência à clindamicina e cefoxitina respectivamente. Poucas Instituições no Brasil, realizam periodicamente teste de sensibilidade a antrimicrobianos para amostras do grupo B. fragilis isoladas de espécimes clínicos. A variação no perfil de sensibilidade entre as espécies do grupo e o alto percentual de resistência à clindamicina e cefoxitina encontrados neste trabalho, reforçam a necessidade do isolamento, caracterização e monitoração do perfil de sensibilidade a antimicrobianos destes microrganismos, pelo menos nos Hospitais Universitários do país.

In Brazilian folk medicine, Lippia sidoides (Ls) and Myracrodruon urundeuva (Mu) have gained popularity and reputation as effective antimicrobial and anti-inflammatory agents. This work aimed to evaluate the effect of topical herbal gel from Ls 0.5% (v/w) and Mu 5% (w/w) in experimental periodontal disease (EPD) in rats. Wistar rats were subjected to ligature placement around the second upper left molars. Animals were treated topically with Ls and/or Mu-based gel, immediately after EPD induction and three times/day for 11 days until the rats were sacrificed (11th day). Saline-based gel was utilized as control for all experiments and doxycycline based gel 10% (w/w) was utilized as reference substance. Animals were weighed daily. Alveolar bone loss was measured as the difference (in millimeters) between the cusp tip and the alveolar bone. The periodontum and the surrounding gingivae were examined at histopathology, as well as the neutrophil influx into the gingivae was assayed using myeloperoxidase activity and cytokine production mainly tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) levels by ELISA method. The local bacterial flora was assessed through culture of the gingival tissue in standard aerobic and anaerobic media. Alveolar bone loss was significantly inhibited by Ls and Mu combined treatment compared to the saline control group. Ls and Mu combined treatment reduced tissue lesion at histopathology, with partial preservation of the periodontum, coupled to decreased myeloperoxidase activity as well as significantly inhibited TNF-alpha and IL-1beta production in gingival tissue compared to the saline control group. Ls and Mu combined treatment also prevented the growth of oral microorganisms and the weight loss. Ls and Mu combined based gel treatment preserved alveolar bone resorption and demonstrated anti-inflammatory and antibacterial activities in experimental periodontitis.