F. J. Gutiérrez Mañero

The ability of four plant growth-promoting rhizobacteria, isolated in a previous study, to induce systemic resistance on Arabidopsis thaliana Col 0 against biotic and abiotic stress was evaluated. All the bacteria enhanced protection against the foliar pathogen Pseudomonas syringae DC3000 and increased plant tolerance to salt stress (NaCl 60 mM). Bacillus sp. strain L81 and Arthrobacter oxidans strain BB1 performed best with a decrease in the disease index of 61.2 and 52.3%, respectively, and a reduction in the mortality due to salt stress of 72.4 and 57.8%, respectively. Additionally, significant differences were found in growth and photosynthesis, again, L81 and BB1 performed best either in normal or under stress conditions. In order to elucidate the pathway elicited by these two strains to induce systemic resistance, experiments with the transgenic line of Arabidopsis thaliana NahG (defective in salicylic acid [SA]) and with the jar1 mutant (defective in jasmonic acid) were carried out. Results showed that the SA-dependent pathway was involved in the defense response induced by strains L81 and BB1. Results from quantitative reverse transcription-polymerase chain reaction analysis of the PR1 gene, related to the SA-dependent pathway and the PDF1.2 gene related to the SA-independent pathway, showed an increased expression of PR1 in BB1-treated plants, confirming involvement of the SA-dependent pathway in the defensive response.

A study of plant defensive systemic responses induced by three plant growth promoting rhizobacteria (PGPR) on Arabidopsis thaliana Col 0 against Pseudomonas syringae pv. tomato DC3000 at the biochemical and transcriptional levels is reported in this paper. All three strains decreased disease severity when applied to A. thaliana prior to pathogen inoculation. At the biochemical level, each of the three strains induced ethylene (ET) when incubated with 1-amino-cyclopropane-1-carboxylic acid, and salicylic acid (SA) production in the plant. Plants treated with each of the three strains were also reduced in salicylic acid production after pathogen challenge compared to untreated controls. This effect was more marked in plants treated with Chryseobacterium balustinum AUR9, the strain most effective in decreasing disease severity. The expression level of PR1, a transcriptional marker of the SA-dependent pathway in C. balustinum AUR9-treated plants, is fourfold that of controls while the expression of PDF1.2, a transcriptional marker for the SA-independent pathway, is not induced. C. balustinum cell wall lipopolysaccharides, being putative bacterial elicitor molecules, are able to reproduce this systemic induction effect at low doses. From these observations, we hypothesize that certain PGPR strains are capable of stimulating different systemic responses in host plants. With C. balustinum AUR9, the SA-dependent pathway is stimulated first, as indicated by increases in SA levels and PR1 expression, followed by induction of the SA-independent pathway, as indicated by the increases in ET concentrations. The effects of both pathways combined with respect to disease suppression appear to be additive.

Two Gram-negative, plant growth-promoting rhizobacteria (PGPRs), denominated as M12 and M14, were classified by 16S rDNA sequencing as Burkholderia graminis species. Both strains were shown to produce a variety of N-acyl-homoserine lactone (AHL) quorum sensing (QS) signalling molecules. The involvement of these molecules in plant growth promotion and the induction of protection against salt stress was examined. AHL production was evaluated in vitro by thin-layer chromatography using AHL biosensors, and the identity of the AHLs produced was determined by liquid chromatography-tandem mass spectrometry. The in situ production of AHLs by M12 and M14 in the rhizosphere of Arabidopsis thaliana plants was detected by co-inoculation with green fluorescent protein-based biosensor strains and confocal laser scanning microscopy. To determine whether plant growth promotion and protection against salt stress were mediated by QS, these PGPRs were assayed on wild-type tomato plants, as well as their corresponding transgenics expressing YenI (short-chain AHL producers) and LasI (long-chain AHL producers). In wild-type tomato plants, only M12 promoted plant growth, and this effect disappeared in both transgenic lines. In contrast, M14 did not promote growth in wild-type tomatoes, but did so in the LasI transgenic line. Resistance to salt stress was induced by M14 in wild-type tomato, but this effect disappeared in both transgenic lines. The strain M12, however, did not induce salt resistance in wild-type tomato, but did so in LasI tomato plants. These results reveal that AHL QS signalling molecules mediate the ability of both PGPR strains M12 and M14 to promote plant growth and to induce protection against salt stress.

The effect of inoculation with PGPR belonging to the genus Bacillus (Bacillus licheniformis CECT 5106 and B. pumilus CECT 5105) in enhancing growth of Pinus pinea plants, and the changes that occur in rhizosphere microbial communities were evaluated. Both Bacillus strains promoted the growth of P. pinea seedlings (probably by gibberellin production), but this biological effect was not found with both strains in combination, which implies a possible competition effect. The introduction of both inoculants caused an alteration in the microbial rhizosphere composition, despite the low levels of inoculants which were found at the end of the assay.

The effects of inoculation with a strain of Bacillus licheniformis on the growth of pepper and tomato were investigated in three experiments, one under seedbed conditions and two under greenhouse production conditions. In the first experiment, the bacterium significantly increased the height of plants and the leaf area in both species and in both cultivars. Effects were greater on pepper than on tomato. In the second experiment, seedlings growing in sand and in hydroponic culture were studied. The number and diameter of tomato fruits produced in sand and in hydroponic medium were increased significantly by inoculation. Treated plants had less disease than non-treated plants. In the third experiment the total weight of pepper harvested from inoculated plants increased significantly with regard to control non-inoculated plants. This strain had considerable colonisation and competitive ability, and it could be used as a biofertiliser or biocontrol agent without altering normal management in greenhouses.

This chapter contains sections titled: Introduction Rhizosphere Microbial Ecology Plant Growth Promoting Rhizobacteria (PGPR) Rhizosphere Microbial Structure Methods to Study the Microbial Structure in the Rhizosphere Ecology and Biodiversity of PGPR Living in the Rhizosphere Diazotrophic PGPR Bacillus Pseudomonas Rhizobia Microbial Activity and Functional Diversity in the Rhizosphere Methods to Study Activity and Functional Diversity in the Rhizosphere Activity and Effect of PGPR in the Rhizosphere Screening Strategies of PGPR Conclusions Prospects References

Hypericum perforatum L. (Guttiferae) appears as an alternative treatment to mild and moderate depression and been traditionally used as a health enhancer based on the phytochemicals hyperforin and hypericin. However, field grown medicinal plants show variable levels of phytopharmaceuticals depending on environmental conditions. Elicitation is a good strategy to trigger secondary metabolism.This study explored the ability of 6 rhizobacterial strains to trigger secondary metabolism in H. perforatum seedlings and molecular elicitors from the most effective strain N5.18 were tested in shoot cultures.Hypericin and pseudohypericin were determined on seedlings and shoot cultures by HPLC. Three putative elicitors from bacterial culture media were assayed in three different concentrations.Strain N5.18 significantly increased hypericin up to 1.2 ppm and pseudohypericin up to 3.4 ppm, over controls (0.3 and 2.5 ppm, respectively) when delivered to seedlings. In shoot cultures, only pseudohypericin was detected (168.9 ppm) and significant increases were observed under the different elicitors, reaching values of 3164.8 ppm with small elicitors in the middle concentration.Secondary metabolism in plants is highly inducible due to its role in plant communication and defense. Our findings demonstrate that some beneficial bacterial strains are able to trigger secondary metabolism in H. perforatum plants when delivered through the roots and bacterial determinants released to culture media are able to reproduce the effect in shoot cultures. Therefore, these elicitors have great potential to enhance phytopharmaceutical production.

<h3>Background</h3> Plasma concentrations of lipoprotein (a) (Lp(a)), a lipoprotein with atherogenic and thrombogenic properties, have a strong genetic basis, although high concentrations of Lp(a) have also been reported in the context of inflammation, as in rheumatoid arthritis (RA). Few studies evaluate the impact of biologic therapies (BT) on Lp(a) in RA, taking into account that with these new therapies a better control of inflammation is achieved. <h3>Objective</h3> The aim of the study was to evaluate the plasma concentrations of Lp(a) in Spanish RA patients on BT attending rheumatology outpatient clinics. <h3>Methods</h3> Baseline analysis of the CARdiovascular in rheuMAtology project, a 10-year prospective study, evaluating the risk of cardiovascular events in RA and other forms of inflammatory arthritis. RA patients were classified according to treatment: no biologic, anti-tumor necrosis factor, anti-interleukin-6 receptor tocilizumab (TCZ), and other biologic (rituximab or abatacept). A model of linear multivariate regression was built in which the dependent variable was Lp(a) concentration and the explanatory variable was BT. The model was adjusted for confounding factors. <h3>Results</h3> Seven hundred and seventy-five RA patients were analyzed. Plasma concentrations of total cholesterol and triglyceride were significantly higher in TCZ-treated patients. Nevertheless, no significant difference in the atherogenic index between TCZ-treated patients and patients without BT was found. After adjusting for confounding factors, patients with BT had lower concentrations of Lp(a) than those without BT; however, only TCZ-treated patients achieved statistically significant differences (β: −0.303, 95% confidence interval: −0.558 to −0.047; <i>P</i> = .02). <h3>Conclusions</h3> RA patients treated with TCZ show lower plasma concentrations of Lp(a) compared with patients without BT.

The effects of five plant growth promoting rhizobacteria on the biological nitrogen fixation (BNF), nodulation, and growth promotion of plants of Lupinus albus cv. Multolupa were investigated. The plants were selected for their capacity to use 1-aminocyclopropane-1-carboxylic acid (ACC) as the sole source of nitrogen. Four strains belonged to the genus Pseudomonas (Luc 1, Luc 2, Luc 3, and Luc 4) and one (Luc 5) belonged to the genus Bacillus. Three patterns of inoculation were examined. In the first pattern, PGPRs were inoculated seven days before being inoculated with B. japonicum. In the second pattern, PGPRs and B. japonicum were co-inoculated, and in the last pattern, PGPRs were inoculated seven days after being inoculated with B. japonicum. The plants were sampled 30 and 45 days after being inoculated with B. japonicum (T1 and T2). In the first pattern of inoculation, Luc 5 significantly increased the biological nitrogen fixation compared to the control at the first sampling time, as did Luc 1 and Luc 4 at the second sampling time. In the second pattern of inoculation, Luc 5 negatively effected the biological nitrogen fixation at both sampling times. In the third pattern of inoculation, all PGPRs caused a decrease in the nitrogen content of the plants compared to the control. The results obtained according to the patterns of inoculation showed that the mechanisms of action of the effects and routes used by Gram-negative and Gram-positive strains were clearly different. Competition between PGPRs and B. japonicum, competition for the niches in the rhizoplane, production of auxins, and induction of systemic resistance (ISR) by the production of siderophores or by lipopolysaccharides present in the outer membrane (LPS) are discussed as probable reasons for the effects observed.

Abstract In order to select potential Plant Growth Promoting Rhizobacteria (PGPRs), a selection of strains from the predominant genera in the rhizosphere of four lupine species, based on genetic divergence criteria, carried out in a previous study, yielded 11 Aureobacterium (Aur), four Cellulomonas (Cell), two Arthrobacter (Arth), two Pseudomonas (Ps), and six Bacillus (Bc) strains. Cell‐free culture filtrates of each bacterium were assayed for effects on germination, growth, and biological nitrogen fixation (BNF) of Lupinus albus L. cv. Multolupa seeds or seedlings. Four (Aur 6, Aur 9, Aur 11, and Cell 1) of the twenty‐five strains assayed promoted germination. Aureobacterium 6 and Aur 9 also increased root surface, total nitrogen content, and BNF. As a result of the screening, and considering all the variables studied, Aur 6 can be considered a plant growth promoting rhizobacterium and is suitable for further field trials in other plants and in different production systems.

Chapter 3 Physiological and Molecular Mechanisms of Plant Growth Promoting Rhizobacteria (PGPR) Beatriz Ramos Solano, Dpto. CC. Ambientales y Recursos Naturales, Facultad de Farmacia, Universidad San Pablo CEU, Urb. Montepríncipe, Ctra. Boadilla del Monte Km 5.3, 28668 Boadilla del Monte, Madrid, SpainSearch for more papers by this authorJorge Barriuso, Dpto. CC. Ambientales y Recursos Naturales, Facultad de Farmacia, Universidad San Pablo CEU, Urb. Montepríncipe, Ctra. Boadilla del Monte Km 5.3, 28668 Boadilla del Monte, Madrid, SpainSearch for more papers by this authorFrancisco J. Gutiérrez Mañero, Dpto. CC. Ambientales y Recursos Naturales, Facultad de Farmacia, Universidad San Pablo CEU, Urb. Montepríncipe, Ctra. Boadilla del Monte Km 5.3, 28668 Boadilla del Monte, Madrid, SpainSearch for more papers by this author Beatriz Ramos Solano, Dpto. CC. Ambientales y Recursos Naturales, Facultad de Farmacia, Universidad San Pablo CEU, Urb. Montepríncipe, Ctra. Boadilla del Monte Km 5.3, 28668 Boadilla del Monte, Madrid, SpainSearch for more papers by this authorJorge Barriuso, Dpto. CC. Ambientales y Recursos Naturales, Facultad de Farmacia, Universidad San Pablo CEU, Urb. Montepríncipe, Ctra. Boadilla del Monte Km 5.3, 28668 Boadilla del Monte, Madrid, SpainSearch for more papers by this authorFrancisco J. Gutiérrez Mañero, Dpto. CC. Ambientales y Recursos Naturales, Facultad de Farmacia, Universidad San Pablo CEU, Urb. Montepríncipe, Ctra. Boadilla del Monte Km 5.3, 28668 Boadilla del Monte, Madrid, SpainSearch for more papers by this author Book Editor(s):Dr. Iqbal Ahmad, Department of Agricultural Microbiology, Faculty of Agricultural Sciences, Aligarh Muslim University, Aligarh 202002, IndiaSearch for more papers by this authorProf. Dr. John Pichtel, Ball State University, Department of Natural Resources and Environmental Management, WQ 103, Muncie, IN 47306, USASearch for more papers by this authorDr. Shamsul Hayat, Aligarh Muslim University, Department of Botany, Aligarh 202002, IndiaSearch for more papers by this author First published: 26 March 2008 https://doi.org/10.1002/9783527621989.ch3Citations: 32 AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinked InRedditWechat Summary This chapter contains sections titled: Introduction PGPR Grouped According to Action Mechanisms PGPR Using Indirect Mechanisms Free Nitrogen-Fixing PGPR Siderophore-Producing PGPR Phosphate-Solubilizing PGPR PGPR Using Direct Mechanisms PGPR that Modify Plant Growth Regulator Levels PGPR that Induce Systemic Resistance Conclusions Future Prospects References Citing Literature Plant‐Bacteria Interactions: Strategies and Techniques to Promote Plant Growth RelatedInformation

In this study, 10 putative plant growth-promoting rhizobacteria (PGPR) were assayed for their ability to improve Pinus pinea growth and mycorrhization.After an inoculation assay, except for two, all strains stimulated plant growth. All bacteria altered rhizosphere microbial communities as revealed by phospholipid fatty acid analysis; associating plant growth promotion with a decrease in biological diversity. Three strains were tested for their ability to enhance pine mycorrhization with wild fungi species. Only strain BB1 increased the total number of mycorrhizal root tips. Mycorrhizas present in the roots of each treatment were identified by ribosomal RNA sequencing and denaturing gradient gel electrophoresis analysis, detecting specificity between mycorrhizal species colonizing the roots and the inoculated PGPR.In conclusion, BB1 appears to be a good candidate to be developed into a biofertilizer directed to enhance pine growth and mycorrhization, which should result in a better establishment rate for plants used in reforestation.This study shows the potential of PGPR to improve fitness of forest tree specie. Moreover, the specificity between the bacteria inoculated and the mycorrhiza that the plant selects involve a potential biotechnological use in production of value-added fungi.

The metabolic capacities of rhizosphere bacteria can depend on intraspecific genetic variability at strain level. We sampled bacteria from the rhizosphere of three populations of four different Lupinus species at two growth stages (flowering: GS1 and fruiting: GS2). Isolates were identified to the genus level by classical biochemical tests. The most abundant genera found were Bacillus, Aureobacterium, Cellulomonas, Pseudomonas, and Arthrobacter. Genetic divergence of rhizobacteria was tested by PCR-RAPDs. The genetic distances were low, with mean values of 37 % for Bacillus, 25 % for Aureobacterium, 46 % for Cellulomonas, 16 % for Pseudomonas, and 23 % for Arthrobacter. Aureobacterium, the most abundant genus, predominated in the rhizosphere of all populations and at both growth stages (GS1 and GS2) of L. angustifolius. The Aureobacterium strains consisted of 11 groups with 90 % similarity indexes. The cluster analysis of these groups shows that strains isolated from different lupin species and sampling times have extraordinary low diversity indexes, or are even identical. This fact, together with the low genetic distance detected in the rhizosphere, reveals a clear specificity in the plant-bacteria interaction. This specificity could be related with several aspects of plant physiology. Genetische Variabilität von Rhizosphärenbakterien, bestimmt über PCR-RAPDs, bei Wildtyppopulationen von vier verschiedenen Lupinus-Spezies Intraspezifische genetische Variabilität kann die metabolischen Leistungen von Rhizosphärenbakterien beeinflussen. Deshalb wurden Bakterien aus der Rhizosphäre dreier Populationen von vier verschiedenen Lupinus-Arten im Blühstadium (GS1) und im fruchttragenden Stadium (GS2) untersucht. Die Identifizierung der isolierten Bakterien erfolgte auf Gattungsniveau mit üblichen biochemischen Tests. Die am häufigsten auftretenden Gattungen waren Bacillus, Aureobacterium, Cellulomonas, Pseudomonas und Arthrobacter. Die genetische Divergenz der Rhizosphärenbakterien wurde anhand von PCR-RAPDs geprüft. Die gefundenen kurzen genetischen Abstände verteilen sich wie folgt auf Bacillus (37 %), Aureobacterium (25 %), Cellulomonas (46 %), Pseudomonas (16 %) und Arthrobacter (23 %). Die häufigste Art Aureobacterium kommt hauptsächlich in der Rhizosphäre aller Populationen und in beiden Wachstumsstadien (GS1 und GS2) von L. angustifolius vor. Die isolierten Bakterien der Gattung Aureobacterium bestanden aus 11 Gruppen mit einer Übereinstimmung von 90 %. Die Cluster-Analyse dieser Gruppen zeigt, dass die isolierten Bakterien, welche unterschiedlichen Lupinenarten und auch Probenahmezeiten zugehören, geringe Unterschiedlichkeitsindices aufweisen, und teilweise sogar identisch sind. Diese Tatsache zeigt, zusammen mit dem niedrigen genetischen Abstand der isolierten Rhizosphärenbakterien, eine klare Spezifität der Wechselwirkung zwischen Pflanze und Bakterien an. Diese Spezifität könnte mit verschiedenen Aspekten der Pflanzenphysiologie verknüpft sein.

Biologic and targeted synthetic disease-modifying antirheumatic drugs (ts/bDMARDs) play a pivotal role in the treatment of rheumatoid arthritis (RA), psoriatic arthritis (PsA), and ankylosing spondylitis (AS). Persistence of therapy provides an index of a drug's overall effectiveness. The objective of the study was to identify factors associated with discontinuation of ts/bDMARDs in a real-world dataset. The study population comprised patients diagnosed with RA, PsA, and AS included in the BIOBADASER registry for whom follow-up data were available until November 2019. Patient features and treatment data were included in the analysis. The Kaplan-Meier method was used to study survival of the different drugs according to the reason for discontinuation. Factors associated with discontinuation were studied using Cox regression models and bivariate and multivariate analyses. P values of less than 0.05 were regarded as statistically significant. The study population comprised 4,752 patients who received a total of 8,377 drugs, of which 4,411 (52.65%) were discontinued. The Kaplan-Meier curves showed that survival for first-line treatment was greater in all 3 groups (p < 0.001). Patients with RA had a greater risk of discontinuation if they were younger (HR, 0.99; 95% CI 0.99-1.00), if they were receiving anti-TNFα agents (HR, 0.61; 95% CI 0.54-0.70), and if they had more comorbid conditions (HR, 1.09; 95% CI 1.00-1.17). Patients with PsA had a higher risk if they were women (HR, 1.36; 95% CI 1.15-1.62) and if they were receiving other ts/bDMARDs (HR, 1.29; 95% CI 1.05-1.59). In patients with AS, risk increased with age (HR, 1.01; 95% CI 1.00-1.02), as did the number of comorbid conditions (HR, 1.27; 95% CI 1.12-1.45). The factors that most affected discontinuation of ts/bDMARDs were line of treatment, age, type of drug, sex, comorbidity and the year of initiation of treatment. The association with these factors differed with each disease, except for first-line treatment, which was associated with a lower risk of discontinuation in all 3 diseases.

The separation and identification of a mixture of organic acids by capillary electrophoresis, with direct detection by UV absorption at a wavelength of 200 nm is reported. A solution containing the following organic acids was studied: oxalic, formic, fumaric, malic, succinic, citric, acetic and lactic acid. Separation was achieved using a fused-silica capillary filled with phosphate buffer (200 mM; pH 6) containing N-acetyl-N,N,N-trimethylammonium bromide (0.5 mM) as cationic surfactant, necessary to modify the electrosmotic flow. The composition of organic acids in exudates of Lupinus luteus was determined by this method. Copyright © 1999 John Wiley & Sons, Ltd.

Cardiovascular (CV) disease is one of the main causes of morbi-mortality in spondyloarthritis (SpA), partially explained by traditional CV risk factors. Information on lipoprotein(a) [Lp(a)], a non-conventional risk factor, in SpA is scarce. In this study we assessed the prevalence of hyperlipoproteinaemia(a) in SpA patients and analysed the possible related factors.A baseline analysis was made of ankylosing spondylitis (AS) and psoriatic arthritis (PsA) patients and controls included in the CARMA project (CARdiovascular in RheuMAtology), a 10-year prospective study evaluating the risk of CV events in chronic inflammatory rheumatic diseases. A multivariate logistic regression model was performed using hyperlipoproteinaemia(a) (Lp(a) >50 mg/dl) as a dependent variable and adjusting for confounding factors.19.2% (95% CI: 16.80-22.05) of the SpA patients [20.7% (95% CI: 16.91-24.82) of those with AS and 17.7% (95% CI: 14.15-21.75) of those with PsA] and 16.7% (95% CI: 13.23-20.86) of the controls had hyperlipoproteinaemia(a) (p=0.326). Adjusting for age and sex, SpA patients were more likely to have hyperlipoproteinaemia(a) than controls (OR: 1.43, 95%CI: 1.00-2.04; p=0.05), especially those with AS (OR: 1.81, 95%CI: 1.18-2.77; p=0.007). In the adjusted model, apolipoprotein B in all patients, non-steroidal anti-inflammatory drugs in AS, and female sex in PsA, were associated with hyperlipoproteinaemia(a). No disease-specific factors associated with hyperlipoproteinaemia(a) were identified.SpA patients show a moderately increased risk of hyperlipoproteinaemia(a) compared to controls, especially those with AS. Lp(a) determination may be of interest to improve the CV risk assessment in SpA patients.

Seasonal changes of bacteria associated with the rhizosphere of Alnus glutinosa (L) Gaertn. were studied in a European alder copse situated in the center of Spain. Bacterial genera were determined following a modification of the Sato et al. (1987) protocol. The role of the bacteria in the nitrogen cycle was determined. The cenotic composition of the European alder rhizosphere alternated between autumn and winter, when Bacillus predominated, and spring and summer, when Pseudomonas predominated. Correlations with different biotic and abiotic parameters are discussed.

The aim of this study was to determine the main factor affecting mineralizing and nitrifying bacteria in a riparian bank, where alder ( Alnus glutinosa L. Gaertn., diazotrophic plant), willow (Salix alba L.) and poplar (Populus × hybrida Moench 1–214) were the main species of vegetation. This was investigated by estimating the most probable number of microorganisms in soil samples from roots collected in spring, summer, autumn, and winter. The numbers of the total microbial population and the ammonifying and proteolytic microorganisms were higher in the plant rhizosphere and rhizoplane than in the shaded soil, litter, and root‐free soil under the trees. Nitrifier densities were the lowest around alder roots. Principal components analysis showed that the main factor affecting microorganism density was the rhizosphere of alder trees. Our results suggest that roots favor microorganism colonization (proteolytic and ammonifying microorganisms) and that roots, particularly alder roots, inhibit nitrifying microorganisms. Keywords: Alnus glutinosa (L.) Gaertn.ecological factorsnitrifying bacteriaN‐mineralizing bacteriaprincipal components analysis

<h3>Background</h3> Persistence to treatment may be used as a surrogate marker for long-term treatment success <h3>Objectives</h3> To assess the probability of persistence on golimumab (GOL) up to 5 years after treatment initiation and the factors associated to longer persistence <h3>Methods</h3> BIOBADASER is the Spanish registry of biological drugs of the Spanish Society of Rheumatology and the Spanish Medicines Agency. A data-base analysis was done in October 2017 on all the patients who had initiated GOL for one of the approved indications (rheumatoid arthritis [RA], axial spondyloarthritis [SpA] or psoriatic arthritis [PsA]). The probability of persistence was calculated with a Kaplan-Meier test. Factors related to persistence were analysed with a Cox-regression model. <h3>Results</h3> 353 patients were included (105 [29.8%] RA, 147 [30.6%] axial SpA and 101 [28.6%] with PsA), mean age 52¹¹ years, 55% women). Median duration of disease at the onset of GOL was 8.0 [2.8–15.0] years. GOL was the first biological drug in 40.1% of the patients, second in 30.1% and third or further biological in 29.8%. Concomitant medication at GOL initiation was methotrexate (MTX) (33.7%), sulphasalazine (SSZ) (6.1%), leflunomide (LEF) (12.7%), steroids (CS) (26.0%). At the last observation on GOL, 32.0% were on MTX, 5.8% on SSZ, 12.8% on LEF, 16.3% on CS. The probability of persistence on GOL since treatment initiation was 85.9% at year 1 (95% CI 81.4–89.5), 73.7% at year 2 (67.1–79.1), 68.5% at year 3 (60.5–75.1), 60.6% (50.2–69.5) at year 4% and 57.1% (44.9–67.5) at year 5. Persistence was similar for RA, axial SpA or PsA patients (p log-rank 0.070), and higher when GOL was used as first biological agent (p log-rank &lt;0.001). As first biological drug the probability of persistence was 94.5% (year 1) and 85.4% (year 2), (insufficient number of cases to assess persistence at year 3 or further). As second biological drug, it was 89.8% (year 1), 75.2% (year 2), 67.4% (year 3) and 59.2% (year 4) and as third biological drug the figures were, respectively, 69.6%, 58.4%, 54.5% and 46.1%. Cox-regression analysis (table 1) showed that the probability of persistence on GOL therapy was higher in first vs second or third biological line patients (Hazard Ratio for discontinuation [HR]: 2.30 [95% CI: 1.16–4.55] for second and 3.92 [2.07–7.39] for third line), and in patients with concomitant MTX (HR discontinuation=0.55 [0.33–0.91]), and lower in those needing CS (HR discontinuation=2.83 [1.72–4.66]). <h3>Conclusions</h3> In patients with RA, axial SpA or PsA, the probability of persistence on GOL after initiation was high. The retention rate was higher both in patients with GOL as first biological drug and in those with concomitant therapy with MTX, and lower in those needing CS. <h3>Acknowledgements</h3> This study was funded by Merck Sharp and Dohme of Spain <h3>Disclosure of Interest</h3> None declared

Esta memoria se basa en la hipotesis del papel fundamental que el sistema microbiano asociado al sistema radical de las plantas tiene sobre el metabolismo vegetal y las consiguientes aplicaciones de los productos derivados del mismo. El sistema rizosferico microbiano, el microbioma rizosferico cumple un papel fundamental para que la planta consiga mejorar sus capacidades de adaptacion a un ambiente cambiante. En el capitulo 2 se exponen de manera enlazadas hipotesis sucesivas que pretenden evidenciar el potencial de aplicacion de las bacterias asociadas a la planta y los distintos enfoques de aplicacion biotecnologicos. En primer lugar, se plantea el uso de la rizosfera como fuente de microorganismos especialmente adaptados a la interaccion del sistema planta/microorganismos. Un sistema en el que la presion selectiva definida por la planta condiciona el tipo de microorganismos, su diversidad y en definitiva la estructura de las comunidades microbianas que se desarrollan en este ecosistema. Sobre la hipotesis de la capacidad de seleccion de microorganismos por la planta, primero se busca una planta que aporte una serie de factores de presionseleccion. La planta se elige en base a criterios filogeneticos y metabolicos (metabolismo secundario muy activo). Nicotiana glauca, es una Solanacea, de la misma familia que especies con gran interes alimentario, como el tomate, Solanum lycopersicum, la patata, Solanum tuberosum o el pimiento, Capsicum annum. Se selecciona esta planta como sujeto de muestreo rizosferico, en busca de un microbioma cultivable y con aplicaciones por sus aportaciones beneficiosas en la interaccion. A continuacion, sobre las casi mil cepas aisladas de la rizosfera, a lo largo de dos anos, en tres suelos de caracteristicas muy diferentes, se realiza un ensayo previo de actividades con potencial para incidir favorablemente sobre la salud de la planta. Las cepas se seleccionan sobre la base de un screening a gran escala en el que se intenta absorber la maxima variabilidad genetica de los microorganismos que se desarrollan en el sistema rizosferico de Nicotiana glauca y pasan a estudiarse por su capacidad para inducir resistencia sistemica y efectos sobre el crecimiento en plantas de tomate, especie elegida como modelo de trabajo...

El desarrollo industrial supone la emision de multiples sustancias contaminantes a nuestro entorno (solidas, liquidas y gaseosas), con el consiguiente deterioro del mismo. En cuanto a las emisiones liquidas, objeto del presente trabajo, la legislacion actual (Ley 10/1993, de la Comunidad de Madrid, sobre vertidos liquidos industriales al sistema integral de saneamiento), es cada vez mas exigente con respecto a parametros quimicos y biologicos (DQO, pH, conductividad, DBO5). En los procesos de descontaminacion se utilizan, en muchas ocasiones, metodos fisico-quimicos pero estos suelen ser perjudiciales para el medio ambiente. Por eso se han desarrollado metodos biologicos, mas respetuosos, que son conocidos como mecanismos de biorremediacion (uso de plantas y/o microorganismos para eliminar la contaminacion existente en un determinado medio). El medio para biorremediar utilizado en este trabajo se conoce como agua de taladrina. La taladrina es una emulsion de agua y aceite (2%) que se utiliza para lubricar y refrigerar la zona de contacto entre las herramientas de corte y los materiales a mecanizar. Pero con el uso, este liquido pierde sus cualidades y se convierte en un residuo (taladrina agotada). La empresa John Deere Iberica S. A. (JDISA), que suministro el liquido a fitorremediar, realiza un tratamiento de la taladrina agotada mediante el sistema del vacudest, obteniendo una reduccion importante de la DQO, desde mas de 25.000 ppm hasta alrededor de 4.000 ppm. Despues de este tratamiento se obtiene un residuo conocido como agua de taladrina. El objetivo del presente trabajo fue desarrollar un sistema biologico capaz de reducir la DQO de las muestras de agua de taladrina procedentes de los procesos de produccion de JDISA. Las muestras estan formadas, principalmente, por hidrocarburos lineales, ramificados, aromaticos y aromaticos policiclicos (fluoreno y pireno), aunque aparecen otros elementos quimicos de diferente naturaleza. Presentan unos valores de DQO superiores a 1.750 ppm (limite legal establecido por la Ley 10/1993 de la Comunidad de Madrid). El desarrollo del sistema biorremediador se dividio en dos partes. En primer lugar se determino la especie vegetal con mayor capacidad para reducir la DQO. Para ello, se utilizaron especies con habitos acuaticos y terrestres y se realizaron diversos ensayos para determinar su resistencia al medio contaminado. La especie que tuvo una mayor resistencia y que era capaz de reducir la DQO por debajo de los limites establecidos por la ley, fue el maiz (Zea mays), con la que se realizaron los ensayos de biorremediacion. Paralelamente se busco un medio en el que las plantas pudiesen enraizar. Este medio tenia que ser biodegradable y no toxico. El esparto resulto ser el soporte de crecimiento vegetal que mas favorecio el enraizamiento de las plantas. En segundo lugar, se aislaron microorganismos de un medio contaminado con hidrocarburos, de las propias taladrinas, del esparto y de un reactor biologico de membrana (R. B. M.). Con estas cepas se desarrollaron sistemas maiz-esparto-microorganismo. Fruto de este trabajo, se solicito y fue concedida la patente, titulada ?Procedimiento para la reduccion de la DQO de efluentes liquidos aceitosos mediante fitorremediacion con maiz-esparto; su aplicacion al tratamiento de taladrinas agotadas? (P201000684). El sistema maiz-esparto fue el mas eficaz en el proceso de fitorremediacion, consiguiendo reducciones del parametro de la DQO en torno a un 60% partiendo de niveles proximos a los 2.000 ppm. Estas reducciones permiten obtener un residuo con estandares de contaminacion por debajo de los limites establecidos por la ley 10/1993 de la Comunidad de Madrid. Solo el sistema de bioaumento desarrollado con un consorcio bacteriano formado por cepas adaptadas a medios contaminados con taladrinas fue capaz de mejorar la fitorremediacion. Paralelamente, se realizaron estudios ultraestructurales, observandose una reduccion del numero de cloroplastos y una fuerte alteracion morfologica de los mismos en las plantas que crecieron en agua de taladrina. En la zona radical no se observo una modificacion de los haces vasculares, lo que podria permitir la translocacion de productos toxicos a la zona aerea. El sistema de fitorremediacion realizado en fases disminuyo el dano causado por el agua de taladrina en las plantas, lo que permitio realizar una segunda fase de fitorremediacion mejorando el rendimiento del proceso. Se observo tambien que la reduccion de la DQO ejercida por este sistema fue mayor que la obtenida por el sistema en continuo. Finalmente, se busco determinar si el sistema de fitorremediacion era capaz de reducir la toxicidad del agua de taladrina. Para ello, se realizo un analisis de perdida de bioluminiscencia con la cianobacteria sensor Anabaena CPB4337, que es muy sensible a la presencia de contaminantes. Se observo que la bioluminiscencia emitida por dicha cianobacteria crecida en las muestras fitorremediadas era 2 veces mayor a la emitida en las muestras iniciales, de manera que el sistema de fitorremediacion no solo es capaz de reducir la DQO sino que a su vez es capaz de disminuir la toxicidad del medio.

Abstract Abstract Microbial communities (phospholipid fatty acid pattern, bacterial growing strategies, eco-physiological index (EPI) and total bacteria counts, as a number of heterotrophic cuhurable bacteria), substrate-induced respiration (SIR), and nitrogen mineralization were studied in three Mediterranean soils at three different depth levels (A, B and C). Soils were experimentally treated with a final concentration of 1000 ppm of trace metals (Cu2+, Zn2+, Al3+, Fe2+, Pb2+, Ni2+, Mn2+, Cr3+ and Cd2+). Soils were stored in 571 plastic containers for one year, and watered with 1001 during this period. Leachate was recovered through a bottom tap. Samples of the three depths were studied. Soil microbial communities showed different effects to other studies presented in the literature, but carried out on non-Mediterranean soils. Dramatic differences were found between treated soils and untreated ones, but not between soils or horizons. the treated soil displayed a decrease in CFUs, SIR N-mineralization and EPI together with a dominance of r-growing strategists. the relative moles percent of several PLFAs, especially 15:0, 16: 1ω7, cy17: 0, br18:0 and 18: 1ω7 decreased because of the pollution of soils, whereas 10Me16, 18:2ω6, cy19:0, i16:0 and br17:0 showed higher values than in untreated soils. This article is part of the following collections: Multiple approaches for assessing the effects of contaminants on the biota

Microbial rhizosphere communities of four different Lupinus species were studied at two plant growth stages in order to determine the dominance of r/K strategists and the pattern of taxa and diversity (Shannon index). Five hundred and seventy-six strains were isolated, determined and classified into eleven different bacterial genera and the Enterobacteriaceae family. Aureobacterium dominated on L. hispanicus and L. angustifolius at both sampling times (GS1: Growth Stage 1 (flowering); GS2: Growth Stage 2 (fruiting)), whereas it only dominated in the rhizosphere of L. albus at GS1 (58.45%). Cellulomonas dominated at GS2 (42.93%). In L. luteus, Bacillus dominated at GS1 (41.21%) and Aureobacterium at GS2 (39.74%). Principal Component Analysis (PCAs) showed that the microbial composition of the rhizosphere of L. albus is the only one that changed considerably from GS1 to GS2. These changes were due to a proportional increase of r strategists at GS2. Sample size (18 to 54 colonies, depending on lupin species) was adequate according to the Shannon index. Despite the fact that diversity between GS1 and GS2 hardly varied in L. albus, L. luteus, and L. hispanicus, the PCAs revealed the largest differences in the microbial structure associated with L. albus. A tentative successional model for the lupin microbial population of the rhizosphere related to its biological cycle, is proposed.

The rhizosphere of Elaeagnus angustifolia L. was sampled monthly during one year and the genera of the isolated bacteria were determined. Bacillus predominated in all seasons. Eight groups of Bacillus were identified according to biochemical tests described in the Bergey's Manual, from which, group VII, with B. mycoides and B. laterosporus as the most likely species to define it, predominated (total year average of 60.5%). In order to determine intrageneric variation, additional tests as resistance to antibiotics and use of different carbon sources were assayed. According to both tests, the Bacillus strains isolated from the rhizosphere of E. angustifolia differed significantly throughout the year. These results suggest the adaptation of rhizospheric microbial communities to environmental conditions, so variable in Mediterranean climates, and to the physiological status of the plant.

<h3>Background</h3> Cardiovascular disease (CVD) is the main cause of mortality and morbidity in patients with spondyloarthritis (SpA), partially explained by traditional CV risk factors (CVRF). Other non-conventional CVRF, probably related to chronic systemic inflammation, may be involved. In this sense, lipoprotein (a) [Lp (a)], an non-conventional risk factor with proatherogenic and thrombogenic properties, could be involved, since it seems to act as an acute-phase reactant, there are few data on this aspect in these patients. <h3>Objectives</h3> To evaluate the prevalence of hyperlipoproteinemia (a) in patients with SpA and analyse the possible related factors. <h3>Methods</h3> Analysis of the baseline visit of patients with ankylosing spondylitis (AS) and psoriatic arthritis (PsA) of the CARMA project (CARdiovascular in Reumatology), a prospective cohort study of 10 years of follow-up, to evaluate the cardiovascular risk in chronic rheumatic inflammatory diseases, including rheumatoid arthritis, AS and PsA, followed in 67 Spanish rheumatology centres. A multivariate logistic regression model was performed, in which the dependent variable was hyperliproteinemia (a), defined as the plasma concentration of lipoprotein (a) [Lp (a)]≥50 mg/dl. Sociodemographic factors and those related to the disease itself have been included as independent variables. <h3>Results</h3> 1459 patients were analysed, 738 with AS and 721 with PsA. Plasma concentrarions of Lp(a) were available in 57.7% of the patients with AS and in 57.1% of the patients with PsA. A 19.2% (95% CI: 16.80–22.05) of the patients with SpA, 20.7% (95% CI: 16.91–24.82) of AS and 17.7% (95% CI: 14.15–21.75) of PsA, respectively, had hyperlipoproteinemia (a), without statistically significant differences with respect to the control group: 16.7% (95% CI: 13.23–20.86; p=0.326). After adjusted for age and sex, only patients with AS were more likely to have hyperlipoproteinemia (a) than the control grup (OR: 1.806, 95% CI: 1177–2.771, p=0.007). In the model adjusted for possible confounding factors, high values of apolipoprotein B in all patients, non-steroidal anti-inflammatories in AS, and sex (men) and kidney disease as comorbidity in PsA, were associated with a higher probability of presenting hyperlipoproteinemia (a). <h3>Conclusions</h3> Patients with AS have a higher percentage of hyperlipoproteinemia (a) compared to the control group. No specific factors of the disease have been identified that are associated with hyperlipoproteinemia (a) in each of the analysed groups. <h3>Disclosure of Interest</h3> None declared

Background: Ustekinumab has been efficacy and safety for psoriatic artritis in clinical trials. Objectives: To assess effectiveness, by means of drug persistence analisys, and safety of ustekinumab in patients with psoriastic arthritis in Biobadaser. Methods: BIOBADASER is the Spanish registry of biological drugs of the Spanish Society of Rheumatology and the Spanish Medicines Agency. We identified patients aged 18 years or more with psoriatic arthritis on Ustekinumab. A descriptive analysis was performed.The persistence of ustekinumab therapy was calculated with a Kaplan-Meier curve and was compared with the persistence of anti-TNF, according to line treatment. Log Rank test was used to establish a comparison. Adverse events occurring with ustekinumab are described according to year treatment. Results: One hundred and twelve patients were on ustekinumab. Most of them were on their second or third line treatment: 53.57% more than one biological therapy (BT), 19.64% second BT, 26.79% were naïve for BT. Most of them were on 45 mg dose: 88.24%. Median duration of disease at Ustekinumab initiation was 10.1 SD 7.2 years; 69.23% had peripheral arthritis; 45.24% had obesity and 39.29% were overweight; 40,6% were on prednisone and 59.82% on DMARD. The cause of discontinuation of treatment was mainly inefficacy (82.61%) and less common an adverse event (6.52%). The probability of persistence of treatment with ustekinumab was 0.83 (95% CI 0.63-0.92) at year 1, 0.79 (0.58-0.90) at year 2 and 0.79 (0.58-0.9) at year 3 when ustekinumab was prescribed as the first line treatment. The persistence decrease when ustekinumab was prescribe as a second and third treatment: being 0.53 (0.27-0.73) the first year, 0.46 (0.22-0.67) the second year and 0.46 (0.22-0.67) as a second line treatment and 0.58 (0.44-0.70) the first year, 0.33 (0.17-0.50) the second year and 0.33 (0.17-0.50) the third year as a third line treatment.The persistence was similar to anti-TNF treatment, according to line treatment. Adverse events were mainly mild (97.83%) and occurred the first year of treatment. Most of the adverse events were classified as “infections and infestations” (36.96%). Conclusion: The persistence of ustekinumab was high, being 83% at the end of the first year on treatment and 79% the second and the third year of treatment. The persistence of ustekinumab was higher when if it was the first line treatment compared as if it was used as the second o third BT option. The persistence of Ustekinumab is similar to the persistence of anti-TNF treatments in all the analyzed treatment lines (no statistically differences were found). Adverse events occurred mainly during the first year treatment. They were mainly mild adverse events and the frequency decreased within the second and third year of treatment. References: [1]Treatment with ustekinumab in a Spanish cohort of patients with psoriasis and psoriatic arthritis in daily clinical practice. Almirall M, Rodriguez J, Mateo L, Carrascosa JM, Notario J, Gallardo F. Clin Rheumatol. 2017 Feb;36(2):439-443; [2] Minimal disease activity in patients with psoriatic arthritis treated with ustekinumab: results from a 24-week real-world study. Napolitano M, Costa L, Caso F, Megna M, Scarpa R, Balato N, Ayala F, Balato A. J Clin Rheumatol. 2018 Oct;24(7):381-384; [3] Minimal Disease Activity and Patient-Acceptable Symptom State in Psoriatic Arthritis: A Real-World Evidence Study With Ustekinumab. Queiro R, Brandy A, Rosado MC, Lorenzo A, Coto P, Carriles C, Alperi M, Ballina J. Actas Dermosifiliogr. 2018 Jun 28; [4] An analysis of Drug Survival, Effectiveness, and Safety in Moderate to Severe Psoriasis Treated With Ustekinumab: An Observational Study of 69 Patients in Routine Clinical Practice. Salgüero Fernández I, Gil MH, Sanz MS, Gullón GR; Disclosure of Interests: None declared

The toxicity of three heavy metals, Cd, Zn and Cu, has been tested in a Mediterranean soil. The soil was incubated (108h) with mixed solutions of those metals before evaluating denitrification and CO2 production, both by gas chromatography. These activities were used as biological indicators of heavy metal toxicity, and compared to non-treated control soil samples. Statistical analyses showed no significant differences in CO2 production between treated and non-treated control soils. The lowest levels of respiration were observed in soils treated with the largest amounts of Zn and Cd. Denitrification increased significantly in soils treated with solutions containing 100 micrograms/ml of Cu and 1000 micrograms/ml of Cd or Zn.

The effect of root nodule extracts from European alder (Alnus glutinosa (L.) Gaertn.) on the rates of amrnonification, nitrification and C02 production was evaluated in three soils of different physico-chemical characteristics. The extracts decreased net ammonification rate (up to a 43% res- pect to water control), probably because of an increase of immobilization rather than an effect of a speciffc inhibitor. The rate of nitrification was increased by the extracts (up to 40% from water con- trol) because of the ammonium present in the extracts. An increase in C02 production showed that the nodule extracts stimulate microbial activity. This depended not only on the amount of organic matter introduced by the extracts but also to some activator of microflora.

espanolNodulos radicales de aliso europeo [Alnusglutinosa (L. )Gaertn. ] fueron mantenidos en condiciones que optimizan el proceso de exudacion (estres hidrico). Se evaluo el efecto de dichos extractos sobre la fijacion libre de nitrogeno (ARA) y desnitrificacion en tres suelos con diferencias significativas en sus caracteristicas fisico-quimicas. Las comparaciones con diferentes controles, muestran que los extractos activan (en un 9. 97%) el ARA y promueven una inhibicion parcial de la desnitrificacion. Los efectos no parecen causados por el aporte de carbono organico o de nitrato por parte de los exudados, sino por algun activador o inhibidor especifico de los procesos considerados. EnglishEuropean alder (Alnus glutinosa [L. ] Gaertn) nodules were maintained under conditions that allow their exudation (water stress). The effect of this nodular extracts colected on free nitrogen fixation (acetylene reduction assay [ARA]) and denitrification, was evaluated in three soils of significativelydifferent physico-chemicalcharacteristics. The comparisons with controls having different concentrations of glucose as carbon source suggest that, in the assayed conditions, the nodular european alder extracts activate ARA (up to 9. 97% respect water control) and produces a partial inhibition of denitrification. The effects do not seem to be due neither to organic carbon nor to nitrate, but rather to specific activator(s) and inhibitor(s) of the processes considered.

Se hace un estudio sobre la influencia que la planta actinorriza myrica gale l. Ejerce sobre la densidad y actividad de los grupos funcionales del ciclo del nitrogeno, diazotroficos, proteoliticos, amonificantes, nitrificantes y desnitrificantes. Tras los resultados de unas experiencias previas realizadas en tres estaciones de muestreo y en invernadero, se estudia con mas detalle dicha influencia en el brezal con m. Gale del valle de la viuda (ciudad real), observando tambien la variacion estacional de la microflora citada bajo el dosel y lejos del mismo. De los resultados obtenidos se deduce que m. Gale estimula la densidad poblacional de los microorganismos diazotrOficos anaerobios, proteoliticos, amonificantes y desnitrificantes y no se aprecian diferencias significativas en los diazotroficos aerobios y nitrificantes. El otono aparece como la epoca mas favorable para el desarrollo microbiano, el invierno, como la mas desfavorable y se observan fluctuaciones claras con respecto a las estaciones del ano en proteoliticos, amonificantes y desnitrificantes. Con respecto a los nutrientes edaficos, se advierte que, bajo la planta, aumenta significativamente la concentracion de carbono organico y las de nitrogeno total y amonio. Ademas, el ph del suelo disminuye.