Effects of desiccation on seed germination, cracking, fungal infection and survival in storage of Sterculia foetida L

Seeds of Sterculia foetida were tested for germination following desiccation and subsequent hermetic storage. Whereas seeds at 10.3% moisture content were intact and provided 98% germination, further desiccation reduced germination substantially. The majority of seed coats had cracked after desiccation to 5.1% moisture content. Ability to germinate was not reduced after 12 months' hermetic storage at 10.3% and 7.3% moisture content at 15 degrees C or -18 degrees C, but was reduced considerably at 5.1%. Fungal infection was detected consistently for cracked seeds in germination tests and they did not germinate. However, almost all embryos extracted from cracked seeds germinated if first disinfected with sodium hypochlorite (1%, 5 minutes). In addition. 80 -100% of disinfected extracted embryos from cracked seeds stored hermetically for 28 d at -18 degrees C or -82 degrees C with 3.3% to 6.0% moisture content, and excised embryos stored in this way, were able to germinate. Hence. failure of the very dry seeds of Sterculia foetida to germinate was not due to embryo death from desiccation but to cracking increasing susceptibility to fungal infection upon rehydration. Cracking was associated negatively and strongly with relative humidity and appears to be a mechanical consequence of substantial differences between the isotherms of whole seeds compared with cotyledons and axes.

Revascularization and periapical repair after endodontic treatment using apical negative pressure irrigation versus conventional irrigation plus triantibiotic intracanal dressing in dogs` teeth with apical periodontitis

Objective. The objective of this study was to evaluate in vivo the revascularization and the apical and periapical repair after endodontic treatment using 2 techniques for root canal disinfection (apical negative pressure irrigation versus apical positive pressure irrigation plus triantibiotic intracanal dressing) in immature dogs` teeth with apical periodontitis. Study design. Two test groups of canals with experimentally induced apical periodontitis were evaluated according to the disinfection technique: Group 1, apical negative pressure irrigation (EndoVac system), and Group 2, apical positive pressure irrigation (conventional irrigation) plus triantibiotic intracanal dressing. In Group 3 (positive control), periapical lesions were induced, but no endodontic treatment was done. Group 4 (negative control) was composed of sound teeth. The animals were killed after 90 days and the maxillas and mandibles were subjected to histological processing. The sections were stained with hematoxylin and eosin and Mallory Trichrome and examined under light microscopy. A description of the apical and periapical features was done and scores were attributed to the following histopathological parameters: newly formed mineralized apical tissue, periapical inflammatory infiltrate, apical periodontal ligament thickness, dentin resorption, and bone tissue resorption. Intergroup comparisons were done by the Kruskal-Wallis and Dunn`s tests (alpha = 0.05). Results. Although statistically significant difference was found only for the inflammatory infiltrate (P < .05), Group 1 presented more exuberant mineralized formations, more structured apical and periapical connective tissue, and a more advanced repair process than Group 2. Conclusion. From the histological observations, sodium hypochlorite irrigation with the EndoVac system can be considered as a promising disinfection protocol in immature teeth with apical periodontitis, suggesting that the use of intracanal antibiotics might not be necessary. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109: 779-787)

Effects of the Association between a Calcium Hydroxide Paste and 0.4% Chlorhexidine on the Development of the Osteogenic Phenotype In Vitro

The present study aimed to evaluate whether the association between a calcium hydroxide paste (Calen paste) and 0.4% chlorhexidine (CHX) affects the development of the osteogenic phenotype in vitro. With rat calvarial osteogenic cell cultures, the following parameters were assayed: cell morphology and viability, alkaline phosphatase activity, total protein content, bone sialoprotein immunolocalization, and mineralized nodule formation. Comparisons were carried out by using the nonparametric Kruskal-Wallis test (level of significance, 5%). The results showed that the association between Calen paste and 0.4% CHX did not affect the development of the osteogenic phenotype. No significant changes were observed in terms of cell shape, cell viability, alkaline phosphatase activity, and the total amount of bone-like nodule formation among control, Calen, or Calen + CHX groups. The strategy to combine Ca(OH)(2) and CHX to promote a desirable synergistic antibacterial effect during endodontic treatment in vivo might not significantly affect osteoblastic cell biology. (J Endod 2008;34:1485-1489)

Effect of a calcium hydroxide-based paste associated to chlorhexidine on RAW 264.7 macrophage cell line culture

Objective. The objective of this study was to evaluate the effect of a calcium hydroxide Ca(OH)(2)-based paste (Calen) associated or not to 0.4% chlorhexidine digluconate (CHX) on RAW 264.7 macrophage cell line culture. Study design. The cell viability (MTT assay), immunostimulating properties (NO dosage), and anti-inflammatory properties (NO, TNF-alpha, and IL-1 alpha dosage) were evaluated after cell exposure to the materials. Data were analyzed statistically by Kruskal-Wallis test at 5% significance level. Results. There was low immunostimulating activity of the Calen paste associated or not to 0.4% CHX in the different materials` concentrations evaluated (P > .05). Anti-inflammatory activity with inhibition of NO and cytokine (TNF-alpha and IL1-alpha) release was observed only with Ca(OH)(2) + CHX at the highest concentration (25 mu g/mL). Conclusion. As the Calen paste associated to 0.4% CHX did not alter cell viability or the immunostimulating and anti-inflammatory properties, the addition of CHX brought no benefits to the Ca(OH)(2)-based paste with regard to the tested parameters. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2008;106:e44-e51)

In vitro antibacterial action of Tetraclean, MTAD and five experimental irrigation solutions

P>Aim To investigate the antibacterial effect of Tetraclean, MTAD and five experimental irrigants using both direct exposure test with planktonic cultures and mixed-species in vitro biofilm model. Methodology Tetraclean, MTAD and five experimental solutions that were modifications of existing formulae including MTAD + 0.01% cetrimide (CTR), MTAD + 0.1% CTR, MTAC-1 (Tween 80 replaced by 0.01% CTR in MTAD), MTAC-2 (Tween 80 replaced by 0.1% CTR) and MTAD-D (MTAD without the Tween 80 and no CTR added) were used as disinfectants in the experiments. In the direct exposure test, a suspension of Enterococcus faecalis was mixed with each of the solutions. After 0.5, 1, 3 and 10 min, an inactivator was added and the number of surviving bacteria was calculated. A mixed-species biofilm from subgingival plaque bacteria was grown in brain heart infusion broth in anaerobic conditions on synthetic hydroxyapatite discs. Two-week-old biofilms were exposed to the solutions for 0.5, 1 and 3 min. The samples were observed by confocal laser scanning microscopy after bacterial viability staining. The scans were quantitatively analysed, and the volume of killed cells of all cells was calculated for each medicament. Results Tetraclean and MTAC-2 (0.1% CTR) killed planktonic E. faecalis in < 30 s. Complete killing of bacteria required 1 min by MTAC-1, 3 min by MTAD + 0.1% CTR and 10 min by MTAD, MTAD-D and MTAD + 0.01% CTR. In the biofilm test, there were significant differences in microbial killing between the different solutions and times of exposure (P < 0.005). MTAC-2 showed the best performance, killing 71% of the biofilm bacteria in 3 min, followed by MTAC-1 and Tetraclean. MTAD and the three MTAD modifications demonstrated the lowest antibacterial activity. Conclusion Tetraclean was more effective than MTAD against E. faecalis in planktonic culture and in mixed-species in vitro biofilm. CTR improved the antimicrobial properties of the solutions, whereas Tween 80 seemed to have a neutral or negative impact on their antimicrobial effectiveness.

High-power diode laser in the disinfection in depth of the root canal dentin

Objective. The objective of this study was to evaluate the disinfection degree of dentine caused by the use of diode laser after biomechanical procedures. Study design. Thirty teeth were sectioned and roots were autoclaved and incubated for 4 weeks with a suspension of Enterococcus faecalis. The specimens were randomly divided into 3 groups (n = 10): G1, instrumented with rotary files, irrigated with 0.5% sodium hypochlorite and 17% EDTA-T, and then irradiated by 830-nm diode laser at 3 W; G2, the same procedures as G1 but without laser irradiation; and G3, irrigation with saline solution (control). Dentin samples of each third were collected with carbide burs and aliquots were sowed to count viable cells. Results. The disinfection degree achieved was 100% in G1 and 98.39% in G2, when compared to the control group (G3). Conclusion. Diode laser irradiation provided increased disinfection of the deep radicular dentin in the parameters and samples tested.

Fungicidal effect of photodynamic therapy against fluconazole-resistant Candida albicans and Candida glabrata

P>Although photodynamic therapy (PDT) has shown great promise for the inactivation of Candida species, its effectiveness against azole-resistant pathogens remains poorly documented. This in vitro study describes the association of Photogem (R) (Photogem, Moscow, Russia) with LED (light emitting diode) light for the photoinactivation of fluconazole-resistant (FR) and American Type Culture Collection (ATCC) strains of Candida albicans and Candida glabrata. Suspensions of each Candida strain were treated with five Photogem (R) concentrations and exposed to four LED light fluences (14, 24, 34 or 50 min of illumination). After incubation (48 h at 37 degrees C), colonies were counted (CFU ml-1). Single-species biofilms were generated on cellulose membrane filters, treated with 25.0 mg l-1 of Photogem (R) and illuminated at 37.5 J cm-2. The biofilms were then disrupted and the viable yeast cells present were determined. Planktonic suspensions of FR strains were effectively killed after PDT. It was observed that the fungicidal effect of PDT was strain-dependent. Significant decreases in biofilm viability were observed for three strains of C. albicans and for two strains of C. glabrata. The results of this investigation demonstrated that although PDT was effective against Candida species, fluconazole-resistant strains showed reduced sensitivity to PDT. Moreover, single-species biofilms were less susceptible to PDT than their planktonic counterparts.

Effect of Fusarium graminearum and infection index on germination and vigor of maize seeds

Patógenos em sementes de milho (Zea mays) causam sérios problemas, como a perda de sua capacidade germinativa. O objetivo do trabalho foi determinar qual o melhor tempo para infecção das sementes de milho com Fusarium graminearum, para posterior avaliação dos danos causados pelo fungo na germinação e vigor das mesmas. As sementes foram colocadas sobre meio de BDA contendo o patógeno e incubadas por 4, 8, 16 e 32 h. Após os respectivos períodos de incubação, estas foram submetidas ao teste de sanidade (papel de filtro), com duas variações, sem e com assepsia superficial, usando hipoclorito de sódio a 1% de cloro ativo, por 3 min. Determinado o melhor tempo para infecção, outras sementes foram infetadas com o patógeno, para realização dos testes de germinação e vigor (envelhecimento acelerado e teste de frio) com uma mistura de sementes sadias (colocadas sobre o meio BDA) e sementes inoculadas, resultando em 0, 20, 40, 60, 80 e 100% de sementes infetadas com o fungo em estudo. Os resultados obtidos mostraram que o período de incubação de 32 h foi suficiente para se obter sementes infetadas. Com relação à germinação, não houve diferenças significativas entre os diferentes níveis de infecção, provavelmente devido ao alto vigor das sementes de milho testadas. Quanto aos testes de vigor, os níveis de infecção diferiram significativamente da testemunha, apesar de não terem diferido entre si.

Avaliação de cultivares de amendoim para resistência a Spodoptera frugiperda

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Processamento mínimo em goiabas 'Paluma' e 'Pedro Sato': 2. Avaliação química, sensorial e microbiológica

Foram utilizados frutos de goiabeiras das cultivares Pedro Sato e Paluma, provenientes de pomar comercial, no estádio de maturação de vez, correspondente à coloração verde-mate e considerados ótimo para o consumo. Os frutos foram inicialmente imersos em solução de hipoclorito de sódio (150mg de cloro.L-1) por 5 minutos, para desinfecção superficial. Pessoas treinadas, utilizando proteção adequada e equipamentos desinfetados descascaram os frutos, cortaram-nos longitudinalmente ao meio e eliminaram a polpa com as sementes, em ambiente a 12°C. Após enxágüe com água clorada (20mg de cloro.L-1) foram embalados em contentores de tereftalato de polietileno (PET) com tampa. Estas unidades foram armazenadas a 3°C por 10 dias. Foram realizadas análises microbiológicas ao longo do período. Determinaram-se quimicamente os conteúdos de lignina, ácido ascórbico, acidez total titulável, sólidos solúveis totais e porcentagem de solubilização das pectinas, bem como as variáveis sensoriais de textura, sabor e preferência. A textura tornou-se mais frágil e os conteúdos de ácido ascórbico se reduziram, ao longo do período de armazenamento, nos produtos de ambas as cultivares. Durante este período houve aumento no conteúdo de lignina e manutenção dos conteúdos de sólidos solúveis totais (SST), acidez total titulável (ATT) e da relação SST/ATT. O produto da cultivar Pedro Sato apresentou menor perda de textura que o da 'Paluma', sendo considerado pelos provadores como o mais saboroso e, portanto, o mais preferido. Devido aos cuidados higiênicos tomados durante o processamento o produto apresentou baixa contagem microbiana (< 10³ UFC.g-1), em todas as avaliações efetuadas.

Avaliação da eficiência da desinfecção de teteiras e dos tetos no processo de ordenha mecânica de vacas

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

Sobrevivência de patógenos associados a sementes de soja armazenadas durante seis meses

Sementes contaminadas ou infectadas no campo podem ter sua sanidade alterada durante o armazenamento, pela perda da viabilidade dos patógenos. O objetivo do presente trabalho foi verificar o efeito do armazenamento na qualidade sanitária de sementes de soja. Variedades de soja BRS 133, MSoy 6101, Conquista e Liderança foram inoculadas, no campo, por pulverização com suspensão de esporos de dois isolados de Colletotrichum dematium var. truncata e Phomopsis sojae. Após a colheita foram realizados testes de sanidade, sem e com desinfestação superficial das sementes com hipoclorito de sódio a 1% por 3 min. As sementes foram armazenadas em câmara fria, por seis meses, e testes de sanidade foram novamente realizados. Os resultados do trabalho permitem concluir que o armazenamento das sementes de soja contaminadas por fungos em câmara fria, por um período de seis meses, diminuiu a incidência de Phmopsis sojae e Colletotricum dematium var. truncata.

Efeito da temperatura no armazenamento de uvas apirênicas minimamente processadas

O trabalho teve como objetivo avaliar a qualidade pós-colheita de três cultivares de uvas de mesa sem semente submetidas ao processamento mínimo e armazenadas sob refrigeração e à temperatura ambiente. Para tanto, foram utilizadas uvas das cultivares BRS Clara, BRS Linda e BRS Morena, produzidas na Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Uva e Vinho/Estação Experimental de Viticultura Tropical, em Jales-SP. Os cachos, depois de higienizados e imersos em água clorada a 200 mg de cloro.L-1 por 5 minutos, foram mantidos em câmara fria, a 12ºC, por 12 h. As bagas foram degranadas e lavadas em solução de álcool a 70%, por 5 segundos. Depois de escorrido o excesso da solução alcoólica, as bagas foram acondicionadas em bandejas de tereftalato de polietileno (PET) transparente com tampa e com capacidade para 500 mL. Cada unidade, contendo 200 g de bagas, foi armazenada a 12±1,8ºC e 24±0,8ºC, por 12 dias. Avaliaram-se, a cada três dias, a perda de massa fresca, a aparência, a coloração e os teores de sólidos solúveis (SS) e de acidez titulável (AT). A temperatura de 12ºC manteve a turgidez, a coloração, as qualidades organoléptica (relação SS/AT) e comercial das bagas das três cultivares testadas, por nove dias, enquanto no armazenamento à temperatura ambiente (24ºC), ocorre perda da qualidade comercial das bagas aos três dias para as cvs. BRS Clara e BRS Linda, e aos seis dias para a cv. BRS Morena.

Processamento mínimo de uvas de mesa sem semente

O trabalho teve como objetivo avaliar os aspectos qualitativos de uvas de mesa apirênicas (sem sementes) quando submetidas ao processamento mínimo e armazenadas sob refrigeração. Para tanto, foram utilizadas uvas da cultivar BRS Morena e da Seleção Avançada nº 8, produzidas na Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Uva e Vinho/Estação Experimental de Viticultura Tropical (da Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) de Jales). Os cachos, depois de higienizados, imersos em água clorada a 300 mg de cloro.L-1 por 5 min., foram mantidos em câmara fria, a 12ºC, por 12 h. Pessoas treinadas e com proteção adequada procederam à degrana dos cachos e ao posterior enxágüe das bagas com água clorada (20 mg.L-1) a 12ºC. Depois de escorrido o excesso de água, as bagas foram acondicionadas em bandejas de tereftalato de polietileno (PET) transparente com tampa e com capacidade para 500 mL. Cada unidade, contendo 200 g de bagas, foi armazenada a 2,5 ± 1ºC e 88% UR por até 36 dias. Avaliaram-se a perda de massa fresca, a evolução da aparência, a coloração e os teores de sólidos solúveis totais (SST), e de acidez titulável (AT). Nas condições do experimento, os produtos minimamente processados da cv. BRS Morena e da Seleção 8 apresentaram baixa perda acumulada de massa fresca (0,16%). O produto da cv. Morena apresentou-se mais escuro (L = 25,04) e mais arroxeado (h° = 332,88) que o da Seleção 8 (L = 29,86 e h° = 345,11), propiciando-lhe melhor qualidade visual. O suco da 'BRS Morena' apresentou maiores teores de SST (22,17 °Brix) e menores para a AT (0,56 %), o que resultou em uma relação SST/AT maior e melhor (39,76) que o da Seleção 8 (18,81). A cv. BRS Morena também apresentou boa manutenção da aparência e, portanto, da qualidade comercial, por 33 dias a 2,5°C, superior ao obtido para a Seleção 8 (24 dias).

Avaliação da produção de biodiesel de microalga Isochrysis galbana via transesterificação in situ

Microalgae are microscopic photosynthetic organisms that grow rapidly and in different environmental conditions due to their simple cellular structure. The cultivation of microalgae is a biological system capable of storing solar energy through the production of organic compounds via photosynthesis, and these species presents growth faster than land plants, enabling higher biomass yield. Thus, it is understood that the cultivation of these photosynthetic mechanisms is part of a relevant proposal, since, when compared to other oil producing raw materials, they have a significantly higher productivity, thus being a raw material able to complete the current demand by biodiesel . The overall aim of the thesis was to obtain biofuel via transesterification process of bio oil from the microalgae Isochrysis galbana. The specific objective was to estimate the use of a photobioreactor at the laboratory level, for the experiments of microalgae growth; evaluating the characteristics of biodiesel from microalgae produced by in situ transesterification process; studying a new route for disinfection of microalgae cultivation, through the use of the chemical agent sodium hypochlorite. The introduction of this new method allowed obtaining the kinetics of the photobioreactor for cultivation, besides getting the biomass needed for processing and analysis of experiments in obtaining biodiesel. The research showed acceptable results for the characteristics observed in the bio oil obtained, which fell within the standards of ANP Resolution No. 14, dated 11.5.2012 - 18.5.2012. Furthermore, it was demonstrated that the photobioreactor designed meet expectations about study culture growth and has contributed largely to the development of the chosen species of microalgae. Thus, it can be seen that the microalgae Isochrysis galbana showed a species with potential for biodiesel production