Susceptibility of planktonic cultures of Streptococcus mutans to photodynamic therapy with a light-emitting diode

The objective of this study was to evaluate the effect of photodynamic therapy with erythrosine and rose bengal using a light-emitting diode (LED) on planktonic cultures of S. mutans. Ten S. mutans strains, including nine clinical strains and one reference strain (ATCC 35688), were used. Suspensions containing 10 6 cells/mL were prepared for each strain and were tested under different experimental conditions: a) LED irradiation in the presence of rose bengal as a photosensitizer (RB+L+); b) LED irradiation in the presence of erythrosine as a photosensitizer (E+L+); c) LED irradiation only (P-L+); d) treatment with rose bengal only (RB+L-); e) treatment with erythrosine only (E+L-); and f) no LED irradiation or photosensitizer treatment, which served as a control group (P-L-). After treatment, the strains were seeded onto BHI agar for determination of the number of colony-forming units (CFU/mL). The results were submitted to analysis of variance and the Tukey test (p ≤ 0.05). The number of CFU/mL was significantly lower in the groups submitted to photodynamic therapy (RB+L+ and E+L+) compared to control (P-L-), with a reduction of 6.86 log 10 in the RB+L+ group and of 5.16 log 10 in the E+L+ group. Photodynamic therapy with rose bengal and erythrosine exerted an antimicrobial effect on all S. mutans strains studied.

Photodynamic inactivation of planktonic cultures and biofilms of Candida albicans mediated by aluminum-chloride-phthalocyanine entrapped in nanoemulsions

New drug delivery systems, such as nanoemulsions (NE), have been developed to allow the use of hydrophobic drugs on the antimicrobial photodynamic therapy. This study evaluated the photodynamic potential of aluminum-chloride- phthalocyanine (ClAlPc) entrapped in cationic and anionic NE to inactivate Candida albicans planktonic cultures and biofilm compared with free ClAlPc. Fungal suspensions were treated with different delivery systems containing ClAlPc and light emitting diode. For planktonic suspensions, colonies were counted and cell metabolism was evaluated by XTT assay. Flow cytometry evaluated cell membrane damage. For biofilms, the metabolic activity was evaluated by XTT and ClAlPc distribution through biofilms was analyzed by confocal laser scanning microscopy (CLSM). Fungal viability was dependent on the delivery system, superficial charge and light dose. Free ClAlPc caused photokilling of the yeast when combined with 100 J cm-2. Cationic NE-ClAlPc reduced significantly both colony counts and cell metabolism (P < 0.05). In addition, cationic NE-ClAlPc and free ClAlPc caused significant damage to the cell membrane (P < 0.05). For the biofilms, cationic NE-ClAlPc reduced cell metabolism by 70%. Anionic NE-ClAlPc did not present antifungal activity. CLSM showed different accumulation on biofilms between the delivery systems. Although NE system showed a lower activity for planktonic culture, cationic NE-ClAlPc showed better results for Candida biofilms. Candida albicans biofilm overview after 30 min of contact with free ClAlPc. This study presents the photodynamic potential of aluminum-chloride-phthalocyanine (ClAlPc) entrapped in cationic and anionic nanoemulsions (NE) to inactivate C. albicans planktonic cultures and biofilm comparing with free ClAlPc. The photodynamic effect was dependent on the delivery system, superficial charge and light dose. Cationic NE-ClAlPc and free ClAlPc caused significant reduction in colony counts, cell metabolism and damage to the cell membrane (P < 0.05). However, only the free ClAlPc was able to cause photokilling of the yeast. The anionic NE-ClAlPc did not present antifungal activity. Although NE system showed a lower activity for planktonic culture, cationic NE-ClAlPc showed better results for Candida biofilms. © 2012 Wiley Periodicals, Inc. Photochemistry and Photobiology © 2012 The American Society of Photobiology.

Effect of different pre-irradiation times on curcumin-mediated photodynamic therapy against planktonic cultures and biofilms of Candida spp

Objectives: The aim of this study was to evaluate the effects of pre-irradiation time (PIT) on curcumin (Cur)-mediated photodynamic therapy (PDT) against planktonic and biofilm cultures of reference strains of Candida albicans, Candida glabrata and Candida dubliniensis. Materials and methods: Suspensions and biofilms of Candida species were maintained in contact with different concentrations of Cur for time intervals of 1, 5, 10 and 20 min before irradiation and LED (light emitting diode) activation. Additional samples were treated only with Cur, without illumination, or only with light, without Cur. Control samples received neither light nor Cur. After PDT, suspensions were plated on Sabouraud Dextrose Agar, while biofilm results were obtained using the XTT-salt reduction method. Confocal Laser Scanning Microscopy (CLSM) observations were performed to supply a better understanding of Cur penetration through the biofilms after 5 and 20 min of contact with the cultures. Results: Different PITs showed no statistical differences in Cur-mediated PDT of Candida spp. cell suspensions. There was complete inactivation of the three Candida species with the association of 20.0 μM Cur after 5, 10 and 20 min of PIT. Biofilm cultures showed significant reduction in cell viability after PDT. In general, the three Candida species evaluated in this study suffered higher reductions in cell viability with the association of 40.0 μM Cur and 20 min of PIT. Additionally, CLSM observations showed different intensities of fluorescence emissions after 5 and 20 min of incubation. Conclusion: Photoinactivation of planktonic cultures was not PIT-dependent. PIT-dependence of the biofilm cultures differed among the species evaluated. Also, CLSM observations confirmed the need of higher time intervals for the Cur to penetrate biofilm structures. © 2012 Elsevier Ltd. All rights reserved.

Two new species of Diaptomidae (Crustacea: Copepoda: Calanoida) from the Neotropical Region (Paraná River)

Two new species of Diaptomidae were found in the Middle Paraná River. The new species are referred to as Diaptomus curvatus sp. nov. and Diaptomus frutosae sp. nov. Important character states in Diaptomus curvatus include: right fifth leg with a well-developed endopod and with the lateral spine of the second exopod segment curved and longer than the segment on which it is inserted, plus the presence of a large dorsal process on urosomite 4 of the male. For Diaptomus frutosae they include: a differentiated hook-like process on segment 20 with chitinous protuberances at base of male right antennule; the last segment of the right leg 5 of male is triangular, and the lateral spine inserted distal to mid-level of this segment. Both species are placed in Diaptomus sensu lato as a temporary placement. http://www.zoobank.org/urn:lsid:zoobank.org:pub:FECDD6C4-C0AC-4043-ADDD-F29539B451F0. © 2013 Copyright Taylor and Francis Group, LLC.

Relaciones entre los rotíferos, copépodos planctónicos ciclopoides y la calidad del agua en dos embalses Brasileños

Planktonic rotifers and cyclopoid copepods were studied in two reservoirs of different trophic states (eutrophic and oligo/mesoeutrophic) in the south of Brazil. During a year, monthly samplings were carried out in three stations in each reservoir. Species richness, frequency and abundance were used to find out useful and indicatives trends of water quality based on these organisms, reinforced by literature data. Species that showed higher differences between reservoirs were chosen. For Rotifera, richness, frequency and abundance of Brachionus were higher in the eutrophic reservoir, but Plationus patulus occurred only in the oligo/mesotrophic reservoir. For copepods, Tropocyclops prasinus dominated in the eutrophic reservoir, but Thermocyclops decipiens, T. minutus, T. inversus and Microcyclops anceps were dominants in the oligo/mesotrophic reservoir. In the canonical correspondence analysis, these species were indicators of the trophic state and were related with chlorophyll-a, total phytoplankton and total phosphorus. The use of these species can be efficient in the studied regions (subtropical/temperate), but comparing with other Brazilian reservoirs of tropical climate, the results could be different. Despite the dominance of T. decipiens over T. minutus, T. inversus has been widely used in Brazil as an indicator of eutrophic waters; in those cases of excessive eutrophication, other species, more rustic, commonly dominate. In the present study, Thermocyclops was dominant in the oligo/mesotrophic reservoir. The dominance of Brachionus for rotifers and Tropocyclops prasinus and Acanthocyclops robustus for copepods were indicative of eutrophic conditions.

Photodynamic potential of curcumin and blue LED against streptococcus mutans in a planktonic culture

Background: The photodynamic therapy (PDT) involves the use of light of specific wavelength to activate a nontoxic photosensitizing agent or dye in the presence of oxygen for eradication of target cells. In dentistry, this therapy is used to suppress the growth of microorganisms involved directly with dental decay and periodontitis process. There are evidences that curcumin dye is able to control microbial activity when illuminated with specific wavelength. The purpose of this study was to evaluate the in vitro efficacy of PDT using curcumin dye (Cur-C) in combination with a blue LED (L) device on a planktonic model of Streptococcus mutans ( S. mutans). Methods: Suspensions (0.5mL) containing S. mutans at 1×107CFUmL-1 were prepared and divided into 4 groups: Group C-L- (control: no treatment and 1 experimental condition), Group C+L- (curcumin at 3 different concentrations: 2000; 4000 and 8000μM and 3 experimental conditions), Group C-L+ (LED at 3 different dosages: 24, 48 and 72Jcm-2 and 3 experimental conditions), and Group C+L+ (PDT group: curcumin at respective concentrations combined to LED dosages and 9 experimental conditions). Samples of each experimental condition were cultured in Petri dishes of BHI agar. Incubation in micro-aerophilia at 37°C for 48h was performed for subsequent visual counting of CFU/mL. Data were transformed into log10 and analyzed by two-way ANOVA and Tukey's test at p<0.05. Results: Group C. +. L+, in specific experimental conditions, demonstrated a log bacterial reduction 70% higher than Group C. -. L-. Both groups C. -. L+ and C. +. L- presented a slight decrease in log bacterial counting. Conclusion: This in vitro method was able to reduce the number of S. mutans in a planktonic suspension. © 2013 Elsevier B.V.

Comparison of the effect of rose bengal- and eosin Y-mediated photodynamic inactivation on planktonic cells and biofilms of Candida albicans

Candida albicans is an opportunistic yeast that can cause oral candidosis through the formation of a biofilm, an important virulence factor that compromises the action of antifungal agents. The objective of this study was to compare the effect of rose bengal (RB)- and eosin Y (EY)-mediated photodynamic inactivation (PDI) using a green light-emitting diode (LED; 532 ± 10 nm) on planktonic cells and biofilms of C. albicans (ATCC 18804). Planktonic cultures were treated with photosensitizers at concentrations ranging from 0.78 to 400 μM, and biofilms were treated with 200 μM of photosensitizers. The number of colony-forming unit per milliliter (CFU/mL) was compared by analysis of variance and Tukey's test (P ≤ 0.05). After treatment, one biofilm specimen of the control and PDI groups were examined by scanning electron microscopy. The photosensitizers (6.25, 25, 50, 200, and 400 μM of EY, and 6.25 μM of RB or higher) significantly reduced the number of CFU/mL in the PDI groups when compared to the control group. With respect to biofilm formation, RB- and EY-mediated PDI promoted reductions of 0.22 log10 and 0.45 log10, respectively. Scanning electron microscopy showed that the two photosensitizers reduced fungal structures. In conclusion, EY- and RB-mediated PDI using LED irradiation significantly reduced C. albicans planktonic cells and biofilms. © 2013 Springer-Verlag London.

Caracterização da comunidade de microcrustáceos e avaliação de potenciais predadores vertebrados e invertebrados em dois reservatórios na região noroeste do Estado de São Paulo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Significant genetic differentiation among populations of Anomalocardia brasiliana (Gmelin, 1791): a bivalve with planktonic larval dispersion

Four Brazilian populations of Anomalocardia brasiliana were tested for mutual genetic homogeneity, using data from 123 sequences of the mtDNA cytochrome oxidase c subunit I gene. A total of 36 haplotypes were identified, those shared being H3 (Canela Island, Prainha and Acupe) and both H5 and H9 (Prainha and Acupe). Haplotype diversity values were high, except for the Camurupim population, whereas nucleotide values were low in all the populations, except for that of Acupe. Only the Prainha population showed a deviation from neutrality and the SSD test did not reject the demographic expansion hypothesis. Fst values showed that the Prainha and Acupe populations represent a single stock, whereas in both the Canela Island and Camurupim stocks, population structures are different and independent. The observed structure at Canela Island may be due to the geographic distance between this population and the remainder. The Camurupim population does not share any haplotype with the remaining populations in northeastern Brazil. The apparent isolation could be due to the rocky barrier located facing the mouth of the Mamanguape River. The results highlight the importance of wide-scale studies to identify and conserve local genetic diversity, especially where migration is restricted.

Avaliação dos parâmetros abióticos e caracterização da comunidade planctônica em um viveiro de cultivo de Litopenaeus vannamei (Boone, 1931) no município de Curuçá - Pará

Este trabalho objetivou avaliar durante um ciclo de cultivo de Litopenaeus vannamei com periodicidade quinzenal de 20/09/08 a 05/12/08 a comunidade planctônica e os parâmetros abióticos em duas estações dentro de um viveiro no município de Curuça, Estado do Pará. Foram medidos transparência, pH, oxigênio dissolvido, salinidade e temperatura, sendo os quatro últimos registrados na superfície e próximo ao fundo e realizadas coletas para o estudo do microfitoplâncton, zooplâncton e clorofila “a”. A temperatura variou de 31,5ºC a 35ºC. O oxigênio dissolvido variou de 4,2 mg/l a 15,5 mg/l. O pH manteve-se ligeiramente alcalino, entre 8,1 e 9,4. A menor salinidade foi 26,9 e a maior, 30 ppm. A transparência diminuiu de 55 cm para 17 cm. Clorofila a teve um mínimo de 2,33mg/m³ e um máximo de 471,34 mg/m³. Foram identificados 95 taxa e Bacillariophyta foi o grupo mais importante, sendo Navicula, Pleurosigma e Nitzschia os principais responsáveis pela sua dominância. A maior densidade registrada para o microfitoplâncton foi de 104.400 org/l no início do cultivo (20/09) e a menor foi 3.600 org/l na última coleta (05/12). A média de diversidade para o fitoplâncton na Estação 01 foi 1,49 bits/ind e na Estação 02, 1,43 bits/ind. Foram identificados 34 taxa zooplanctônicos, sendo Copepoda o grupo mais importante e Acartia lilljeborgi, Euterpina acutifrons, Oithona hebes, Oithona oswaldocruzzi e Parvocalanus crassirostris os principais responsáveis pela sua dominância. A maior densidade registrada para o zooplâncton foi de 162.000 org/m³ no início do cultivo (20/09) e a menor foi 375 org/m³ no dia 05/11. A diversidade também foi baixa tendo médias de 1,34 bits/ind e 1,10 bits/ind nas estações 01 e 02, respectivamente. Entre as principais conclusões: a comunidade microfitoplanctônica foi dominada pelas diatomáceas, sendo os principais gêneros responsáveis por esta dominância: Pleurosigma, Nitzschia e Navicula e a divisão Bacillariophyta foi o grupo mais importante tanto em termos de riqueza quanto de densidade; a classe dinophyceae revelou estar melhor adaptados em águas mais claras; os copépodos foram dominantes, sendo Acartia lilljeborgi, Oithona hebes, Oithona oswaldocruzzi, Parvocalanus crassirostris e Euterpina acutifrons as espécies que mais contribuíram para esta dominância; Clorofila “a” respondeu aos maiores aportes de ração durante o cultivo, aumentando com o tempo; as variáveis físico-químicas que sofreram influência do cultivo, variando significativamente ao longo do tempo foram: pH, oxigênio dissolvido e transparência e o viveiro investigado foi considerado homogêneo avaliando sua profundidade e área.

Estimating cyclopoid copepod species richness and geographical distribution (Crustacea) across a large hydrographical basin: comparing between samples from water column (plankton) and macrophyte stands

Species richness and geographical distribution of Cyclopoida freshwater copepods were analyzed along the La Plata River basin. Ninety-six samples were taken from 24 sampling sites, twelve sites for zooplankton in open waters and twelve sites for zooplankton within macrophyte stands, including reservoirs and lotic stretches. There were, on average, three species per sample in the plankton compared to five per sample in macrophytes. Six species were exclusive to the plankton, 10 to macrophyte stands, and 17 were common to both. Only one species was found in similar proportions in plankton and macrophytes, while five species were widely found in plankton, and thirteen in macrophytes. The distinction between species from open water zooplankton and macrophytes was supported by nonmetric multidimensional analysis. There was no distinct pattern of endemicity within the basin, and double sampling contributes to this result. This lack of sub-regional faunal differentiation is in accordance with other studies that have shown that cyclopoids generally have wide geographical distribution in the Neotropics and that some species there are cosmopolitan. This contrasts with other freshwater copepods such as Calanoida and some Harpacticoida. We conclude that sampling plankton and macrophytes together provided a more accurate estimate of the richness and geographical distribution of these organisms than sampling in either one of those zones alone.

Differential gene expression in Acidithiobacillus ferrooxidans LR planktonic and attached cells in the presence of chalcopyrite

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Trends in planktonic copepod diversity in reservoirs and lotic stretches in a large river basin in South America

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

Ecological aspects and sexual maturity of a southwestern Atlantic population of the planktonic shrimp Lucifer faxoni (Decapoda: Sergestoidea)

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

Antimicrobial activity of root canal irrigants associated with cetrimide against biofilm and planktonic Enterococcus Faecalis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)