Comparison between classical and molecular (FISH and PCR) methods for Lactobacillus spp. detection in clinical samples

Lactobacillus species constitute the main beneficial bacteria in our body by inhibiting the growth from pathogenic microorganisms. Fluorescence In Situ Hybridization (FISH) is an ideal method for cultivation-independent detection of microorganisms in microbial communities or clinical samples. Therefore, the current aims of this research are to identify and discriminate Lactobacillus spp. contained in clinical samples by the use of PNA-FISH methodology. In spite this method is proved to be useful to visualize target cells in natural habitats, it wasn't possible to find a Lactobacillus spp. 16S conservative region that allowed an unique and efficient identification in clinical samples. To overcome this problem, we used morphological visualization to differentiate Lactobacillus genus from another relative genera of the same Lactobacillaceae order. In addition, we also needed to overcome some methodological limitations, such as minimizing probe penetration problems and increasing hybridization efficiencies. As a result, we investigated the effect of different pre-treatment procedures of the exopolymer cell walls prior to the hybridization step, such as, several types of fixation compounds (paraformaldehyde and ethanol percentages), buffer steps and enzymatic (lysozyme and protease) pre-treatment. Furthermore, we modified PNA FISH protocol in several steps, for instance, hybridization and washing steps. In resume, the use of PNA probe specific for Lactobacillus spp. in situ hybridization by fluorescence microscopy could be perfectly used to study the complex and spatial organization of vaginal microbial samples. To conclude, we validate Lactobacillus spp. PNA probe by FISH to quantify and characterize in mixed microbiologic populations present in clinical samples

Fluorescence in situ hybridization method using peptide nucleic acid probes for rapid detection of Lactobacillus and Gardnerella spp

Background: Bacterial vaginosis (BV) is a common vaginal infection occurring in women of reproductive age. It is widely accepted that the microbial switch from normal microflora to BV is characterized by a decrease in vaginal colonization by Lactobacillus species together with an increase of Gardnerella vaginalis and other anaerobes. Our goal was to develop and optimize a novel Peptide Nucleic Acid (PNA) Fluorescence in situ Hybridization assay (PNA FISH) for the detection of Lactobacillus spp. and G. vaginalis in mixed samples. Results: Therefore, we evaluated and validated two specific PNA probes by using 36 representative Lactobacillus strains, 22 representative G. vaginalis strains and 27 other taxonomically related or pathogenic bacterial strains commonly found in vaginal samples. The probes were also tested at different concentrations of G. vaginalis and Lactobacillus species in vitro, in the presence of a HeLa cell line. Specificity and sensitivity of the PNA probes were found to be 98.0% (95% confidence interval (CI), from 87.8 to 99.9%) and 100% (95% CI, from 88.0 to 100.0%), for Lactobacillus spp.; and 100% (95% CI, from 92.8 to 100%) and 100% (95% CI, from 81.5 to 100.0%) for G. vaginalis. Moreover, the probes were evaluated in mixed samples mimicking women with BV or normal vaginal microflora, demonstrating efficiency and applicability of our PNA FISH. Conclusions: This quick method accurately detects Lactobacillus spp. and G. vaginalis species in mixed samples, thus enabling efficient evaluation of the two bacterial groups, most frequently encountered in the vagina.This work was supported by European Union funds (FEDER/COMPETE) and by national funds (FCT) under the project with reference FCOMP-01-0124-FEDER-008991 (PTDC/BIA-MIC/098228/2008). AM acknowledges the FCT individual fellowship - SFRH/BD/62375/2009)

Erratum to: 36th International Symposium on Intensive Care and Emergency Medicine

[This corrects the article DOI: 10.1186/s13054-016-1208-6.]

O balneário pré-romano de Braga

Dá-se a conhecer os resultados obtidos com a escavação do balneário da Idade do Ferro, encontrado em Braga.info:eu-repo/semantics/publishedVersio

Atlantic sea surface temperatures estimated from planktonic foraminifera off the Iberian Margin over the last 40Ka BP

Sea surface temperatures (SSTs) were reconstructed from planktonic foraminiferal assemblages in two sediment cores off the NW Iberian Continental Margin for the last 40 kyr. Results of this work were compared with published SSTs estimated in six cores from the Iberian Margin and the delta O-18 data from NGRIP ice core of Greenland. Longitudinal and latitudinal SST changes were identified in several intervals related to alterations in the hydrographic conditions off the Iberian Peninsula, namely the influence of the upwelling system, the intensity of Paleo-Portugal and Paleo-Azores Currents, and the positions of the Polar and Arctic Fronts. During the Holocene, the SSTs were quite stable along the west Iberian Margin when compared with the last glaciation. However, during the Last Glacial Maximum (LGM) SSTs were not much lower than during the Holocene. The Portugal Current, as an eastward descending branch of the North Atlantic Drift, should have been quite vigorous in transporting warm water to the west Iberian Margin, which could have moderated the SSTs during the LGM. However, our results suggest a much higher instability during the LGM than previously reported and much lower SSTs at 42 degrees N. The lowest SSTs during the last 40 ka were recorded during the Heinrich Stadials (HS) and the Younger Dryas (YD). During the HS, SSTs at the northwest Iberian Margin were lower than today by about 10 to 16 degrees C as a result of the invasion of cool and low-salinity water masses and the increased transport of icebergs from the northern seas. The Polar Front was persistently situated at 42 degrees N for similar to 3 ka during the HS1, and a comparable present-day Arctic Front should have been present up to the latitude of 40 degrees N during the HS2, HS3, and HS4 off the west Iberian Peninsula. The HS were recorded all along the west Iberian Margin. However, an overall trend of increasing of mean SSTs from north to south of similar to 8 degrees C during the HS1, of similar to 10 degrees C during the HS2b and HS3, of similar to 11 degrees C during the HS2a, and of similar to 4 degrees C during the HS4 was recorded. These gradients also were present during the YD of similar to 9 degrees C and the LGM of similar to 4 degrees C and may be related to the strong influence of the Paleo-Azores Current in southern areas

Atlantic sea surface temperatures estimated from planktonic foraminifera off the Iberian Margin over the last 40 Ka BP

Sea surface temperatures (SSTs)were reconstructed fromplanktonic foraminiferal assemblages in two sediment cores off the NWIberian Continental Margin for the last 40 kyr. Results of this work were compared with published SSTs estimated in six cores from the Iberian Margin and the δ18O data from NGRIP ice core of Greenland. Longitudinal and latitudinal SST changes were identified in several intervals related to alterations in the hydrographic conditions off the Iberian Peninsula, namely the influence of the upwelling system, the intensity of Paleo-Portugal and Paleo-Azores Currents, and the positions of the Polar and Arctic Fronts. During the Holocene, the SSTswere quite stable along thewest IberianMarginwhen comparedwith the last glaciation. However, during the Last Glacial Maximum(LGM) SSTswere notmuch lower than during the Holocene. The Portugal Current, as an eastward descending branch of the North Atlantic Drift, should have been quite vigorous in transporting warm water to the west Iberian Margin, which could have moderated the SSTs during the LGM. However, our results suggest a much higher instability during the LGMthan previously reported and much lower SSTs at 42°N. The lowest SSTs during the last 40 ka were recorded during the Heinrich Stadials (HS) and the Younger Dryas (YD). During the HS, SSTs at the northwest Iberian Margin were lower than today by about 10 to 16 °C as a result of the invasion of cool and low-salinity water masses and the increased transport of icebergs from the northern seas. The Polar Front was persistently situated at 42°N for ~3 ka during the HS1, and a comparable present-day Arctic Front should have been present up to the latitude of 40°Nduring the HS2, HS3, andHS4 off thewest Iberian Peninsula. The HS were recorded all along the west Iberian Margin. However, an overall trend of increasing of mean SSTs from north to south of ~8 °C during the HS1, of ~10 °C during the HS2b and HS3, of ~11 °C during the HS2a, and of ~4 °C during the HS4 was recorded. These gradients also were present during the YD of ~9 °C and the LGM of ~4 °C and may be related to the strong influence of the Paleo-Azores Current in southern areas

Arquitetura eclética no Brasil: o cenário da modernização

A autora acentua o significado social e cultural do Ecletismo, além do domínio da arquitetura e do nivel aos fenômenos estilísticos. Este quadro lhe permite entender, de um ponto de vista histórico, suas tendências cenográficas, o gosto pela magnificência, a expressividade e a monumentaliaade, assim como os vínculos com o passado. São então apontados alguns traços específicos do Ecletismo no Brasil, que tem entre seus ingredientes uma atitude anti-colonial. O movimento neoclássicoé incluído como manifestação tardia do Ecletismo, que sobreviverá até o estabelecimento de uma nova relação com a modernidade - mediada pela tecnologia