Abstract Haemonchus contortus is an important parasite in sheep and the abuse of anti-helminthics has resulted in a rapid resistance. Little is known about the abomasal microbiota in sheep and the relation to local parasitism. This study aimed to identify bacteria in the abomasum of sheep, through the abomasal content, abomasal mucosa, and adult H. contortus parasites, in high and low parasitism. Eight sheep naturally infected with H. contortus were classified as high (n = 4) and low infection (n = 4). Samples of abomasal contents, abomasal mucosa, and adult H. contortus parasites were collected. Samples were pooled to form six groups, three in each level of parasitism: high infection/abomasal contents (HC), high infection/mucosa (HM), high infection/parasites (HP); low infection/abomasal contents (LC), low infection/mucosa (LM), and low infection/parasites (LP). Molecular identification of bacteria was performed by the amplification and sequencing of the 16S rRNA bacterial gene. A similar distribution of phyla was observed between pools HC/LC and HP/LP. For pools HM/LM, there was a significant difference (p = 0.01) in the proportion of phyla observed. The bacterial phyla predominant for libraries HC/LC were Firmicutes (82% and 62%) and Bacteroidetes (10.4% and 17.6%); for HM/LM, Firmicutes (76.9% and 56%) and Proteobacteria (10.2% and 38.4%); and for HP/LP, Proteobacteria (42.8% and 55%) and Firmicutes (31.6% and 40%). We observed differences about the samples analyzed, suggesting that there are various bacterial communities closely associated with the different materials analyzed, even from the close environment. antihelminthics anti helminthics resistance parasitism content mucosa H n 4 4. . collected groups infectionabomasal HC, HC , (HC) infectionmucosa HM, HM (HM) infectionparasites HP (HP) LC, LC (LC) LM, LM (LM) LP. LP (LP) S gene HCLC HPLP HP/LP HMLM HM/LM p 0.01 001 0 01 82% 82 (82 62% 62 10.4% 104 10 (10.4 17.6% 176 17 6 17.6%) 76.9% 769 76 9 (76.9 56% 56 10.2% 102 2 (10.2 38.4% 384 38 38.4%) 42.8% 428 42 8 (42.8 55% 55 31.6% 316 31 (31.6 40%. 40 40% 40%) analyzed environment (HC (HM (HP (LC (LM (LP 0.0 00 (8 10.4 1 (10. 17.6 76.9 7 (76. 5 10.2 38.4 3 42.8 (42. 31.6 (31. 0. ( 10. (10 17. 76. (76 38. 42. (42 31. (31 (1 (7 (4 (3

Abstract The combination of nutritional and chemical factors in honey and pollen produced by Tetragonisca angustula (jataí) reveals a unique ecosystem that is conducive to developing microorganisms. However, there is a lack of information regarding the microbial species grown in the nest pots of this stingless bee species. The objectives of this study were to count, isolate and identify the microbiota associated with the pots of honey and pollen. The samples were collected from three jataí beehives. Microbiological analyses showed that the average total aerobic mesophiles ranged between 3.78 and 3.71 log CFU/g; lactic bacteria were 3.59 and 3.56 log CFU/g; and the yeast was 4.41 and 4.66 log CFU/g for the honey and pollen, respectively. There were four gram-positive cocci bacteria (Staphylococcus saprophyticus subsp. bovis, S. vitulinus, S. kloosii and S. pasteuri); two spore-forming gram-positive bacteria (Bacillus pumilus and B. thuringiensis); and one gram-negative bacteria (Pantoe agglomerans). One lactic bacteria (Leuconostoc mesenteroides subsp. mesenteroides) was also identified. Three yeast species were isolated (Starmerella meliponinorum, Candida magnoliae and Zygosaccharomyces bailli). Bacillus was the main bacteria in jataí honey and pollen. Z. bailli was the dominant yeast in the pollen, and S. meliponinorum was the dominant yeast in the honey. T. angustula bees and their products (honey and pollen) are ecosystems that provide sustainable environmental and technological integration. They also provide a selection of microorganisms that can be applied in the food, cosmetic and pharmaceutical industries.

This study evaluated the impact of different concentrations of glyphosate (Rondup®) on planktonic and biofilm growth of P. aeruginosa. Aerobic and anaerobic cultures of P. aeruginosa ATCC®15442 inoculated in MHB + glyphosate (0.845 ppm, 1.690 ppm, 8.45 ppm, 16.90 ppm, 84.50 ppm, 169 ppm, 845 ppm, and 1690 ppm) and cultured in normoxia and anoxia, following their OD560nm every hour for 24 h. Biofilms of adapted cells were formed in the presence of glyphosate (0.845 to 1690 ppm) in normoxia and anoxia for 36 h. Glyphosate at concentrations higher than 84.5 ppm reduces the cell density of planktonic aerobic cultures (p < 0.05). However, these same concentrations favor the planktonic anaerobic growth (p < 0.05). On the other hand, the herbicide favors a slight growth of biofilms in a concentration-dependent manner up to 84.5 ppm (p > 0.05), and more pronounced over 169 ppm. Anaerobic biofilms have their growth more readily favored (p < 0.05), regardless of concentration. In a concentration-dependent manner, glyphosate interferes with the growth ability of P. aeruginosa ATCC®15442.

This study was evaluated the clonal diversity of Streptococcus mutans in caries-free and caries-active subjects using MLEE. Strains from caries-free subjects were grouped in a single taxon. Unrooted dendrogram showed that different strains clustered in four different clades, also showed that more than one clonal type can be found in a same individual.

The genetic diversity of C. albicans oral isolates from 75 healthy schoolchildren from eight schools located in different geographic areas of Piracicaba city, São Paulo state, Brazil, was established using isoenzymes marker (Multilocus Enzyme Electrophoresis - MLEE) and cluster analysis. Patterns of monoclonal and polyclonal oral colonization by C. albicans within and between groups of schoolchildren were identified. However, significant divergence between the observed and the expected genotypic frequencies (Hardy-Weinberg equilibrium test) was not detected in the geographically adjacent groups, suggesting the hypothesis that populations of healthy schoolchildren do not correspond to the selection factor (differential survival) of strains. Two highly polymorphic and distantly genetically related taxa (A and B) were identified within the total population of yeasts, each contained subgroups (A1, A2, A3, A4, B1 and B2) and clusters of moderately related strains (from I to X), suggesting the existence of strains restricted or not to certain groups of geographically limited, healthy students. However, the coexistence of identical strains in healthy schoolchildren from the same school (geographically related) reinforces the hypothesis of oral transmission, where the sources of propagation could be explored. Furthermore, this could also be used in current and retrospective analyses of C. albicans isolated from immunocompetent and immunocompromised people, in order to detect commensal or potentially pathogenic yeast groups, predominantly in candidiasis, and in the development of strategies to prevent transmission or human propagation.

OBJETIVO: Este estudo avaliou as propriedades físicas de dois cimentos MTA disponíveis comercialmente. METODOLOGIA: A resistência a compressão (CS) dos materiais avaliados foi realizada após 21 horas e 14 dias de imersão em água. A avaliação da radiopacidade (RD) dos materiais foi mensurada em função de espessura de alumínio. Com relação à propriedade de tempo de presa (ST), tanto os tempos de presa inicial e final foram mensurados. A solubilidade foi calculada em função da percentagem de massa perdida após armazenamento em água. Os valores de pH foram mensurados em três diferentes tempos (inicial, 1 e 24 horas) de armazenamento em água. Todos os testes foram realizados de acordo com a norma ISO 6876-2:2001. RESULTADOS: Ambos os materiais apresentaram valores de CS e pH em acordo com os valores da norma ISO. Os dois cimentos apresentaram RD superiores ao limite de 3 mm de alumínio. Ambos os materiais mostraram resultados de SB inferiores ao limite de 3%. ST e pH também estão de acordo com a norma ISO 6876-2:2001. Adicionalmente foram avaliadas as superfícies dos materiais por MEV, e ambos apresentaram fases estruturais amorfas e cristalinas. CONCLUSÃO: Concluiu-se que ambos os materiais avaliados estão de acordo com as normas ISO, permitindo o seu uso como material de preenchimento de canais radiculares.

PURPOSE: This study evaluated some physical properties of two commercial MTA cements regarding it use as root canal filling material. METHODS: The compression strength (CS) of the evaluated materials was performed after 21 hours or 14 days of water immersion. The materials' radiopacity (RD) was measured according to aluminum thickness. Regarding setting time (ST), both initial and final times were measured. The solubility (SB) was calculated according to percentage of lost mass after water storage. The pH measurement was performed in three different times (initial, 1 and 24 hours) of water storage. All tests were performed according to ISO 6876-2:2001. RESULTS: Both cements showed that CS and pH were as in accordance with ISO standard. The two cements exhibited RD values higher than the equivalent limit of 3 mm-aluminum. Both materials exhibited SB values within the recommended limit of 3%. ST and pH also are in accordance with ISO 6876-2:2001. Additional SEM surface evaluation showed both amorphous and crystalline phase for both cements. CONCLUSION: It is possible to conclude that all evaluated materials are in accordance with ISO standard, allowing its use as root canal filling material.

O teste de redução do 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) tem sido utilizado para mensurar o desenvolvimento de biofilmes de Candida. Contudo, a reação de XTT é dependente da atividade celular e o seu uso para biofilmes maduros pode ser questionado, considerando que diferentes camadas celulares têm atividade metabólica diferenciadas. O objetivo deste estudo foi avaliar se a adição de glicose à formula de XTT diminuiria a variabilidade na mensuração da atividade metabólica. Biofilmes de Candida albicans ATCC 90028 com tempos de crescimento de 24, 48 e 72 h foram utilizados. Para avaliar o melhor tempo de incubação do XTT, este foi mantido a temperatura de 37 °C, em tempos de 90 180 e 270 min. A fórmula padrão do teste XTT (controle) foi modificada com a adição de 50, 100 e 200 mM de glicose para os grupos experimentais. Os melhores resultados para a incubação foi observado com tempo de 180 min e para a suplementação de glicose à concentração de 200 mM (p<0.001). Concluiu-se que a incubação de 180 min utilizando a suplementação de 200 mM de glicose apresenta resultados de atividade metabólica celular com a menor variação para o estudo de biofilmes de Candida albicans.

2, 3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) reduction assay has been used to study Candida biofilm formation. However, considering that the XTT reduction assay is dependent on cell activity, its use for evaluating mature biofilms may lead to inaccuracies since biofilm bottom cell layers tend to be relatively quiescent at later stages of biofilm formation. The aim of this study was to improve XTT reduction assay by adding glucose supplements to the standard XTT formulation. Candida albicans ATCC 90028 was used to form 24-, 48- and 72-h biofilms. The oxidative activity at 90, 180 and 270 min of incubation was evaluated. The control consisted of standard XTT formulation without glucose supplements, and was modified by the addition of 50, 100 and 200 mM of glucose. The XTT assay with 200 mM glucose showed more accurate and consistent readings correlating with biofilm development at 24, 48 and 72 h. Biofilm growth yield after 180 min incubation, when evaluated with the 200 mM glucose supplemented XTT, produced the most consistent readings on repetitive testing. It may be concluded that glucose supplementation of XTT could minimize variation and produce more accurate data for the XTT assay.

O objetivo deste trabalho foi avaliar a concentração do peróxido de carbamida manipulado em diferentes farmácias de manipulação. Foram utilizados géis clareadores manipulados em farmácias especializadas e um industrializado (grupo controle) (n = 20) com análise de concentração imediata. A concentração do peróxido de carbamida foi obtida por titulometria e os resultados foram submetidos a análise estatística pelo teste de Kruskal-Wallis. Como resultado, o agente clareador (controle) e um dos produtos manipulados em farmácia apresentaram as melhores médias de concentração, próximas a 16%. Pode-se concluir que a concentração do peróxido de carbamida manipulado e dos industrializados apresentaram valores de concentração diferentes de 16%.

The aim of this work was to evaluate the concentration of carbamide peroxide compounded at different dispensing pharmacies. Immediate concentration analysis was made of bleaching gels dispensed by specialized pharmacies, and of a commercially available gel (control group) (n = 20). The carbamide peroxide concentration was determined by titration and the results were analyzed statistically by the Kruskal-Wallis test. The commercial bleaching agent (control group) and one of the gels from the pharmacies presented the best mean concentration values, close to 16%. In conclusion, the concentration of the manipulated and industrialized carbamide peroxide gels presented concentration values differing from 16%.

Vinte e uma cepas de Streptococcus mutans foram agrupadas pela eletroforese de enzimas codificadas por multilocus (MLEE). Seis isoenzimas apresentaram forte poder discriminatório (M1P, MPI, PLP, NSP, GOT e LAP). A MLEE é uma técnica robusta que pode ser empregada no estudo da diversidade clonal de cepas de S. mutans, em estudos epidemiológicos.

Twenty-one Streptococcus mutans strains were clustered by Multilocus Enzyme Electrophoresis (MLEE). Six isoenzymes showed strong infra-specific discriminatory power (M1P, MPI, PLP, NSP, GOT, and LAP). MLEE is a robust technique that may be used to explore clonal diversity of S. mutans isolates in epidemiological surveys.

The microbiota from the uniforms of 31 professionals from the general intensive care unit was analyzed. The samples were collected in duplicate at the beginning and at the end of the work period. Total viable counts of microorganisms were determined; there was a significant increase in the counts at the end of the period, when compared with those obtained at the beginning. No significant difference was observed between the first and second counts obtained from the cuffs. However, differences were observed for the samples from the abdominal region. Among the isolated pathogens 11/18 were Staphylococcus aureus, 2/18 were Acinetobacter baumannii, 2/18 were Klebsiela pneumoniae and 1/18 were Serratia rubidae. Some of these isolates were multi-resistant to antibiotics. Emphasis should be placed on reducing the spread of these pathogens in the hospital, making sure that biosafety protocols are followed by the staff.

O objetivo da presente pesquisa foi avaliar os graus de polimorfismos protéicos entre setenta e cinco linhagens de C. albicans isoladas da cavidade oral de crianças saudáveis provenientes de cinco categorias socioeconômicas e oito escolas (particulares e públicas) do município de Piracicaba, Estado de São Paulo, a fim de identificar subespécies de C. albicans e suas similaridades em grupos de populações infantis e estabelecer suas possíveis rotas de disseminação. Culturas celulares foram desenvolvidas em meio YEPD, coletadas por centrifugação e lavadas com solução salina gelada. As proteínas celulares totais foram extraídas por rompimento celular usando pérolas de vidro e submetidas à técnica de SDS-PAGE. Após a eletroforese, as bandas de proteínas foram coradas com Coomassie-blue e analisadas pelo conjunto de programas estatísticos NTSYS-pc versão 1.70. Matriz de similaridade e dendrograma foram gerados, pela aplicação do coeficiente de similaridade de Dice e do algoritmo UPGMA, respectivamente, os quais permitiram avaliar os graus de polimorfismo ou similaridade intra-específico, baseados nos padrões eletroforéticos de proteínas totais de isolados orais de C. albicans. Um total de 13 principais fenons (grupos) foi analisado de acordo com suas características homogêneas (categoria socioeconômica e/ou escola idênticas) e heterogêneas (categorias socioeconômicas e/ou escolas diferentes). Com relação ao aspecto epidemiológico socioeconômico, as composições dos grupos mostraram alta similaridade (0.788 < S D < 1.0) entre algumas linhagens de C. albicans isoladas de crianças saudáveis independentemente de suas camadas socioeconômicas (alta, média e baixa). Isolados de alta similaridade não foram encontrados nas cavidades orais de crianças saudáveis pertencentes às camadas sociais A e D, B e C, ou C e E. Isto pode ser explicado pela ausência de uma rota de disseminação entre estas crianças. Geograficamente, algumas crianças saudáveis entre escolas idênticas e diferentes (particulares e públicas) também são portadoras de linhagens semelhantes, mas tal similaridade não foi encontrada entre outros de determinadas escolas. Esses dados podem refletir uma rota de disseminação restrita destes microrganismos em alguns grupos de escolares saudáveis, a qual pode ser dependente da categoria socioeconômica ou local geográfico de cada criança. Em contraste à alta similaridade, a baixa similaridade ou alto grau de polimorfismo (0.499 < S D < 0.788) dos perfis protéicos foi demonstrado em 23 (30,6%) isolados orais de C. albicans. Considerando o aspecto epidemiológico social, 42,1%, 41,7%, 26,6%, 23,5% e 16,7% foram isolados de crianças provenientes das categorias socioeconômicas A, D, C, B e E, respectivamente, e geograficamente, 63,6%, 50%, 33,3%, 33,3%, 30%, 25% e 14,3% foram isolados de crianças provenientes das escolas LAE, MA, CS, AV, HF, FMC e MEP, respectivamente. Tais resultados sugerem um maior grau de polimorfismo entre algumas linhagens isoladas de crianças saudáveis independentemente de suas camadas sociais ou locais geográficos. Estudos complementares envolvendo escolares saudáveis e seus familiares, professores, serventes, hábitos nutricionais e de higiene deverão ser realizados a fim de estabelecer as fontes de tais padrões de colonização em grupos de populações de crianças saudáveis. Os perfis de proteínas totais obtidos por SDS-PAGE associados com análise numérica computadorizada podem proporcionar critérios adicionais para os estudos epidemiológicos e taxonômicos de C. albicans.

The aim of this research was to evaluate the protein polymorphism degree among seventy-five C. albicans strains from healthy children oral cavities of five socioeconomic categories from eight schools (private and public) in Piracicaba city, São Paulo State, in order to identify C. albicans subspecies and their similarities in infantile population groups and to establish their possible dissemination route. Cell cultures were grown in YEPD medium, collected by centrifugation, and washed with cold saline solution. The whole-cell proteins were extracted by cell disruption, using glass beads and submitted to SDS-PAGE technique. After electrophoresis, the protein bands were stained with Coomassie-blue and analyzed by statistics package NTSYS-pc version 1.70 software. Similarity matrix and dendrogram were generated by using the Dice similarity coefficient and UPGMA algorithm, respectively, which made it possible to evaluate the similarity or intra-specific polymorphism degrees, based on whole-cell protein fingerprinting of C. albicans oral isolates. A total of 13 major phenons (clusters) were analyzed, according to their homogeneous (socioeconomic category and/or same school) and heterogeneous (distinct socioeconomic categories and/or schools) characteristics. Regarding to the social epidemiological aspect, the cluster composition showed higher similarities (0.788 < S D < 1.0) among C. albicans strains isolated from healthy children independent of their socioeconomic bases (high, medium, or low). Isolates of high similarity were not found in oral cavities from healthy children of social stratum A and D, B and D, or C and E. This may be explained by an absence of a dissemination route among these children. Geographically, some healthy children among identical and different schools (private and public) also are carriers of similar strains but such similarity was not found among other isolates from children from certain schools. These data may reflect a restricted dissemination route of these microorganisms in some groups of healthy scholars, which may be dependent of either socioeconomic categories or geographic site of each child. In contrast to the higher similarity, the lower similarity or higher polymorphism degree (0.499 < S D < 0.788) of protein profiles was shown in 23 (30.6%) C. albicans oral isolates. Considering the social epidemiological aspect, 42.1%, 41.7%, 26.6%, 23.5%, and 16.7% were isolates from children concerning to socioeconomic categories A, D, C, B, and E, respectively, and geographically, 63.6%, 50%, 33.3%, 33.3%, 30%, 25%, and 14.3% were isolates from children from schools LAE (Liceu Colégio Albert Einstein), MA (E.E.P.S.G. "Prof. Elias de Melo Ayres"), CS (E.E.P.G. "Prof. Carlos Sodero"), AV (Alphaville), HF (E.E.P.S.G. "Honorato Faustino), FMC (E.E.P.G. "Prof. Francisco Mariano da Costa"), and MEP (E.E.P.S.G. "Prof. Manasses Ephraim Pereira), respectively. Such results suggest a higher protein polymorphism degree among some strains isolated from healthy children independent of their socioeconomic strata or geographic sites. Complementary studies, involving healthy students and their families, teachers, servants, hygiene and nutritional habits must be done in order to establish the sources of such colonization patterns in population groups of healthy children. The whole-cell protein profile obtained by SDS-PAGE associated with computer-assisted numerical analysis may provide additional criteria for the taxonomic and epidemiological studies of C. albicans.

Duzentos e trinta e nove crianças brasileiras foram estudadas, distribuídas em cinco categorias socioeconômicas distintas (A a E). As amostras de saliva foram analisadas avaliando-se o fluxo salivar, pH, capacidade tampão e parâmetros microbianos. Os resultados mostraram Candida em 47,3% da amostra total. A espécie mais encontrada foi C. albicans em todas as categorias socioeconômicas seguidas por C. tropicalis, C. krusei e C. parapsilosis. A saliva e os parâmetros de investigacão clínicos como a proporcão de secreção, capacidade tampão e CFU/ml de Candida não mostraram nenhuma correlação estatística entre estes parâmetros. A freqüência de Candida não diferiu substancialmente entre os grupos; no entanto elas foram mais acentuadas entre as categorias B e C. Entre todas as espécies, C. albicans foi a mais prevalente. Somente 5% das amostras mostraram mais de uma espécie e revelaram a presença de C. albicans associada com C. tropicalis, C. Parapsilosis ou C. krusei. Foi possível detectar uma correlação significante entre os índices de cárie e as categorias socioeconômicas. As categorias A e E mostraram índices de cárie aumentados; no entanto, a amostra da população foi considerada de risco de cárie baixo. Não havia nenhuma correlação positiva entre Candida e risco de cárie.

Two hundred and thirty-nine (239) Brazilian children, distributed into five distinct socioeconomic categories (A to E) were studied. Saliva samples were analyzed as to flow rate, pH, buffer capacity and microbial parameters. The results revealed the presence of Candida spp. in 47.3% of the samples. The most commonly isolated species was C. albicans, in all socioeconomic categories, followed by C. tropicalis, C. krusei and C. parapsilosis. There was no statistical correlation between secretion rate, buffer capacity and Candida spp. CFU/ml. The prevalence of Candida spp. did not differ substantially among the groups; however the microorganisms were more detected in categories B and C. Among all species, C. albicans was the most prevalent. Only 5% of the sample presented more than one species - C. albicans associated with C. tropicalis, C. parapsilosis or C. krusei. It was possible to detect a significant correlation between caries indices and the socioeconomic categories. All categories presented increased caries indices; however the studied population was considered of low caries risk. There was no positive correlation between the presence of Candida and caries risk in the analyzed population.