Identification of Sinorhizobium (Ensifer) medicae based on a specific genomic sequence unveiled by M13-PCR fingerprinting

A collection of nodule isolates from Medicago polymorpha obtained from southern and central Portugal was evaluated by M13-PCR fingerprinting and hierarchical cluster analysis. Several genomic clusters were obtained which, by 16S rRNA gene sequencing of selected representatives, were shown to be associated with particular taxonomic groups of rhizobia and other soil bacteria. The method provided a clear separation between rhizobia and co-isolated non-symbiotic soil contaminants. Ten M13-PCR groups were assigned to Sinorhizobium (Ensifer) medicae and included all isolates responsible for the formation of nitrogen-fixing nodules upon re-inoculation of M. polymorpha test-plants. In addition, enterobacterial repetitive intergenic consensus (ERIC)-PCR fingerprinting indicated a high genomic heterogeneity within the major M13-PCR clusters of S. medicae isolates. Based on nucleotide sequence data of an M13-PCR amplicon of ca. 1500 bp, observed only in S. medicae isolates and spanning locus Smed_3707 to Smed_3709 from the pSMED01 plasmid sequence of S. medicae WSM419 genome’s sequence, a pair of PCR primers was designed and used for direct PCR amplification of a 1399-bp sequence within this fragment. Additional in silico and in vitro experiments, as well as phylogenetic analysis, confirmed the specificity of this primer combination and therefore the reliability of this approach in the prompt identification of S. medicae isolates and their distinction from other soil bacteria. [Int Microbiol 2009; 12(4):215-225

Proceso asistencial integrado de esclerosis lateral amiotrófica

O proceso asistencial integrado da esclerose lateral amiotrófica, elaborouse co obxectivo de crear un proceso de traballo común en todas as áreas para facilitar a asistencia sanitaria ás persoas diagnosticadas desta enfermidade. Establécense actuacións como o asesoramento continuo, as consultas en acto único, a coordinación asistencial, tanto entre especialidades como coa atención primaria e a coordinación administrativa do sistema socio sanitario. Neste proceso participaron profesionais das diferentes áreas sanitarias especialistas en neuroloxía, endocrinoloxía, neumoloxía, psicoloxía clínica, rehabilitación, traballo social e hospitalización a domicilioEl proceso asistencial integrado de la esclerosis lateral amiotrófica, se elaboró con el objetivo de crear un proceso de trabajo común en todas las áreas para facilitar la asistencia sanitaria a las personas diagnosticadas de esta enfermedad. Se establecen actuaciones como el asesoramiento continuo, las consultas en acto único, la coordinación asistencial, tanto entre especialidades como con la atención primaria y la coordinación administrativa del sistema socio sanitario. En este proceso participaron profesionales de las diferentes áreas sanitarias especialistas en neurología, endocrinología, neumología, psicología clínica, rehabilitación, trabajo social y hospitalización a domicili

Photoprotective Bioactivity Present in a Unique Marine Bacteria Collection from Portuguese Deep Sea Hydrothermal Vents

Abstract: Interesting biological activities have been found for numerous marine compounds. In fact, screening of phylogenetically diverse marine microorganisms from extreme environments revealed to be a rational approach for the discovery of novel molecules with relevant bioactivities for industries such as pharmaceutical and cosmeceutical. Nevertheless, marine sources deliverables are still far from the expectations and new extreme sources of microbes should be explored. In this work, a marine prokaryotic collection from four Mid-Atlantic Ridge (MAR) deep sea hydrothermal vents near the Azores Islands, Portugal, was created, characterized and tested for its photoprotective capacity. Within 246 isolates, a polyphasic approach, using chemotaxonomic and molecular typing methods, identified 23-related clusters of phenetically similar isolates with high indexes of diversity. Interestingly, 16S rRNA gene sequencing suggested the presence of 43 % new prokaryotic species. A sub-set of 139 isolates of the prokaryotic collection was selected for biotechnological exploitation with 484 bacterial extracts prepared in a sustainable upscalling manner. 22 % of the extracts showed an industriall

MLVA16 typing of Portuguese human and animal Brucella melitensis and Brucella abortus isolates.

To investigate the epidemiological relationship of isolates from different Portuguese geographical regions and to assess the diversity among isolates, the MLVA16(Orsay) assay (panels 1, 2A and 2B) was performed with a collection of 126 Brucella melitensis (46 human and 80 animal isolates) and 157 B. abortus field isolates, seven vaccine strains and the representative reference strains of each species. The MLVA16(Orsay) showed a similar high discriminatory power (HGDI 0.972 and 0.902) for both species but panel 1 and 2A markers displayed higher diversity (HGDI 0.693) in B. abortus compared to B. melitensis isolates (HGDI 0.342). The B. melitensis population belong to the "Americas" (17%) and "East Mediterranean" (83%) groups. No isolate belonged to the "West Mediterranean" group. Eighty-five percent of the human isolates (39 in 46) fit in the "East-Mediterranean" group where a single lineage known as MLVA11 genotype 116 is responsible for the vast majority of Brucella infections in humans. B. abortus isolates formed a consistent group with bv1 and bv3 isolates in different clusters. Four MLVA11 genotypes were observed for the first time in isolates from S. Jorge and Terceira islands from Azores. From the collection of isolates analysed in this study we conclude that MLVA16(Orsay) provided a clear view of Brucella spp. population, confirming epidemiological linkage in outbreak investigations. In particular, it suggests recent and ongoing colonisation of Portugal with one B. melitensis lineage usually associated with East Mediterranean countries

<i>Brucella</i> strains used in this work.

<p>A, Alentejo; ALG, Algarve; BI, Beira Interior; BL, Beira Litoral; EDM, Entre Douro e Minho; RO, Ribatejo e Oeste; TM, Trás-os-Montes; ATCC, American Type Culture Collection; CITA, Centro de Investigación y Tecnología Agro Alimentaria del Gobierno de Aragón; NA, Not Applicable.</p

Cluster analysis for 170 <i>B. abortus</i> strains based on the data set of MLVA16_Orsay.

<p>In the columns, the following data for isolates are indicated: genotype numbering (No.), strain identification (ID), biovar (Bv.), year of isolation (Year), geographic region (Region), Panel1 and MLVA11 (Panels1+2A) genotypes corresponding to each isolate in the Brucella2010 database for each set of loci. The colour code reflects the Portuguese regions: Portugal mainland, yellow; Azores, light blue; S. Miguel, dark green; S. Jorge, orange; Terceira, green.A total of 40 genotypes were observed. <b>A</b>, Alentejo; <b>BI</b>, Beira Interior; <b>BL</b>, Beira Litoral; <b>EDM</b>, Entre Douro e Minho; <b>RO</b>, Ribatejo e Oeste; <b>TM</b>, Trás-os-Montes.</p

Minimum Spanning Tree analysis of published <i>B. abortus</i> isolates using the MLVA11 data.

<p>The data were recovered from the published <i>Brucella</i>2010 MLVA database. Colour codes are associated with Portuguese <i>B. abortus</i> genotypes: Portugal mainland, yellow; Azores, light blue; S. Miguel, dark green; S. Jorge, orange; Terceira, green. Circles in white represent genotypes from other countries and previously published in Brucella2010 MLVA database. The numbers represent the MLVA11genotypes found in this study. The size of the shapes indicates the number of strains described in each genotype. Each of the circles showing more than one colour includes the Portuguese genotypes and others previously published.</p

Minimum Spanning Tree analysis of published <i>B. melitensis</i> isolates (excluding vaccine strains) using the MLVA11 data.

<p>Colour codes are associated with the main <i>B. melitensis</i> MLVA groups. The analysis was performed using all isolates from this study and typing data from more than one thousand isolates previously published <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042514#pone.0042514-Alvarez1" target="_blank">[19]</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042514#pone.0042514-Kili1" target="_blank">[22]</a>, Brucella2010 database hosted at <a href="http://mlva.u-psud.fr" target="_blank">http://mlva.u-psud.fr</a>]. Portuguese isolates are shown in yellow. The <i>B. melitensis</i> population is included in “American” (17%) and “East-Mediterranean” (83%) groups. Thirty nine human isolates (85%; 39 out of 46) are MLVA11 genotype 116 usually associated with the “East-Mediterranean” <i>B. melitensis</i> strains and seven isolates (15%) are included in the “American” group. The numbers represent the MLVA11 genotypes found in this study. The size of the shapes indicates the number of strains described in each genotype. Each of the circles showing yellow and dark blue or red colors included the Portuguese genotype (yellow) and the genotypes found in “East-Mediterranean” (dark blue) or “American” (red) <i>B. melitensis</i> isolates.</p

Distribution of animal <i>B. melitensis</i> and <i>B. abortus</i> isolates among Portugal regions.

<p>Distribution of animal <i>B. melitensis</i> and <i>B. abortus</i> isolates among Portugal regions.</p