MALDI-TOF Mass Spectrometry Is a Fast and Reliable Platform for Identification and Ecological Studies of Species from Family Rhizobiaceae

Family Rhizobiaceae includes fast growing bacteria currently arranged into three genera, Rhizobium, Ensifer and Shinella, that contain pathogenic, symbiotic and saprophytic species. The identification of these species is not possible on the basis of physiological or biochemical traits and should be based on sequencing of several genes. Therefore alternative methods are necessary for rapid and reliable identification of members from family Rhizobiaceae. In this work we evaluated the suitability of Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF MS) for this purpose. Firstly, we evaluated the capability of this methodology to differentiate among species of family Rhizobiaceae including those closely related and then we extended the database of MALDI Biotyper 2.0 including the type strains of 56 species from genera Rhizobium, Ensifer and Shinella. Secondly, we evaluated the identification potential of this methodology by using several strains isolated from different sources previously identified on the basis of their rrs, recA and atpD gene sequences. The 100% of these strains were correctly identified showing that MALDI-TOF MS is an excellent tool for identification of fast growing rhizobia applicable to large populations of isolates in ecological and taxonomic studies

An active tectonic field for CO2 storage management: the Hontomín onshore case study (Spain)

One of the concerns of underground CO2 onshore storage is the triggering of induced seismicity and fault reactivation by the pore pressure increasing. Hence, a comprehensive analysis of the tectonic parameters involved in the storage rock formation is mandatory for safety management operations. Unquestionably, active faults and seal faults depicting the storage bulk are relevant parameters to be considered. However, there is a lack of analysis of the active tectonic strain field affecting these faults during the CO2 storage monitoring. The advantage of reconstructing the tectonic field is the possibility to determine the strain trajectories and describing the fault patterns affecting the reservoir rock. In this work, we adapt a methodology of systematic geostructural analysis to underground CO2 storage, based on the calculation of the strain field from kinematics indicators on the fault planes (ey and ex for the maximum and minimum horizontal shortening, respectively). This methodology is based on a statistical analysis of individual strain tensor solutions obtained from fresh outcrops from the Triassic to the Miocene. Consequently, we have collected 447 fault data in 32 field stations located within a 20 km radius. The understanding of the fault sets’ role for underground fluid circulation can also be established, helping further analysis of CO2 leakage and seepage. We have applied this methodology to Hontomín onshore CO2 storage facilities (central Spain). The geology of the area and the number of high-quality outcrops made this site a good candidate for studying the strain field from kinematics fault analysis. The results indicate a strike-slip tectonic regime with maximum horizontal shortening with a 160 and 50◦ E trend for the local regime, which activates NE–SW strike-slip faults. A regional extensional tectonic field was also recognized with a N–S trend, which activates N–S extensional faults, and NNE–SSW and NNW– SSE strike-slip faults, measured in the Cretaceous limestone on top of the Hontomín facilities. Monitoring these faults within the reservoir is suggested in addition to the possibility of obtaining a focal mechanism solutions for microearthquakes (M < 3)This work has been partially supported by the European Project ENOS: ENabling Onshore CO2 Storage in Europe, H2020 Project ID: 653718 and the Spanish project 3GEO, CGL2017-83931-C3-2-P, MICIU-FEDE

Influencia de la castración y del nivel energético de la dieta sobre el crecimiento y composición corporal del borrego pelibuey

EI presente trabajo se llevó a cabo con el fin de conocer el comportamiento productivo del borrego Pelibuey castrado alimentado con niveles crecientes de energía en la dieta, teniendo como punto de comparación el comportamiento de animales enteros

Plants Probiotics as a Tool to Produce Highly Functional Fruits: The Case of Phyllobacterium and Vitamin C in Strawberries

10 páginas, 1 tabla, 1 figuraThe increasing interest in the preservation of the environment and the health of consumers is changing production methods and food consumption habits. Functional foods are increasingly demanded by consumers because they contain bioactive compounds involved in health protection. In this sense biofertilization using plant probiotics is a reliable alternative to the use of chemical fertilizers, but there are few studies about the effects of plant probiotics on the yield of functional fruits and, especially, on the content of bioactive compounds. In the present work we reported that a strain of genus Phyllobacterium able to produce biofilms and to colonize strawberry roots is able to increase the yield and quality of strawberry plants. In addition, the fruits from plants inoculated with this strain have significantly higher content in vitamin C, one of the most interesting bioactive compounds in strawberries. Therefore the use of selected plant probiotics benefits the environment and human health without agronomical losses, allowing the production of highly functional foods.This work was granted by “Junta de Castilla y León” (Regional Government, Grant SA183A11-2) and MINECO (Central Government, Grant AGL2011-29227). Paula García-Fraile is recipient of a postdoctoral researcher contract from Academy of Sciences of the Czech Republic. José David Flores-Félix was supported by a fellowship of Salamanca University. Marta Marcos García was supported by a fellowship of Fundación Miguel Casado San José. Luís R. Silva is grateful to the financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundação para a Ciência e Tecnologia) through project Pest-C/EQB/LA0006/2013 and from the European Union (FEDER funds) under the framework of QREN through Project NORTE-07-0124-FEDER- 000069.Peer reviewe

Implementación de sistemas miniaturizados para la identificación de bacterias lácticas

Memoria ID-257. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2013-2014

Implementación de diferentes tipos de sistemas miniaturizados para la identificación de levaduras que intervienen en la producción de probióticos

Memoria ID-0188. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2014-2015

Defining the Rhizobium leguminosarum Species Complex

Bacteria currently included in Rhizobium leguminosarum are too diverse to be considered a single species, so we can refer to this as a species complex (the Rlc). We have found 429 publicly available genome sequences that fall within the Rlc and these show that the Rlc is a distinct entity, well separated from other species in the genus. Its sister taxon is R. anhuiense. We constructed a phylogeny based on concatenated sequences of 120 universal (core) genes, and calculated pairwise average nucleotide identity (ANI) between all genomes. From these analyses, we concluded that the Rlc includes 18 distinct genospecies, plus 7 unique strains that are not placed in these genospecies. Each genospecies is separated by a distinct gap in ANI values, usually at approximately 96% ANI, implying that it is a ‘natural’ unit. Five of the genospecies include the type strains of named species: R. laguerreae, R. sophorae, R. ruizarguesonis, “R. indicum” and R. leguminosarum itself. The 16S ribosomal RNA sequence is remarkably diverse within the Rlc, but does not distinguish the genospecies. Partial sequences of housekeeping genes, which have frequently been used to characterize isolate collections, can mostly be assigned unambiguously to a genospecies, but alleles within a genospecies do not always form a clade, so single genes are not a reliable guide to the true phylogeny of the strains. We conclude that access to a large number of genome sequences is a powerful tool for characterizing the diversity of bacteria, and that taxonomic conclusions should be based on all available genome sequences, not just those of type strains

Defining the Rhizobium leguminosarum Species Complex

Bacteria currently included in Rhizobium leguminosarum are too diverse to be considered a single species, so we can refer to this as a species complex (the Rlc). We have found 429 publicly available genome sequences that fall within the Rlc and these show that the Rlc is a distinct entity, well separated from other species in the genus. Its sister taxon is R. anhuiense. We constructed a phylogeny based on concatenated sequences of 120 universal (core) genes, and calculated pairwise average nucleotide identity (ANI) between all genomes. From these analyses, we concluded that the Rlc includes 18 distinct genospecies, plus 7 unique strains that are not placed in these genospecies. Each genospecies is separated by a distinct gap in ANI values, usually at approximately 96% ANI, implying that it is a ‘natural’ unit. Five of the genospecies include the type strains of named species: R. laguerreae, R. sophorae, R. ruizarguesonis, “R. indicum” and R. leguminosarum itself. The 16S ribosomal RNA sequence is remarkably diverse within the Rlc, but does not distinguish the genospecies. Partial sequences of housekeeping genes, which have frequently been used to characterize isolate collections, can mostly be assigned unambiguously to a genospecies, but alleles within a genospecies do not always form a clade, so single genes are not a reliable guide to the true phylogeny of the strains. We conclude that access to a large number of genome sequences is a powerful tool for characterizing the diversity of bacteria, and that taxonomic conclusions should be based on all available genome sequences, not just those of type strains