Study of Bacterial Community Composition and Correlation of Environmental Variables in Rambla Salada, a Hypersaline Environment in South-Eastern Spain

We studied the bacterial community in Rambla Salada in three different sampling sites and in three different seasons and the effect of salinity, oxygen, and pH. All sites samples had high diversity and richness (Rr > 30). The diversity indexes and the analysis of dendrograms obtained by DGGE fingerprint after applying Pearson’s and Dice’s coefficient showed a strong influence of sampling season. The Pareto-Lorenz (PL) curves and Fo analysis indicated that the microbial communities were balanced and despite the changing environmental conditions, they can preserve their functionality. The main phyla detected by DGGE were Bacteroidetes (39.73%), Proteobacteria (28.43%), Firmicutes (8.23%), and Cyanobacteria (5.14%). The majority of the sequences corresponding to uncultured bacteria belonged to Bacteroidetes phylum. Within Proteobacteria, the main genera detected were Halothiobacillus and Roseovarius. The environmental factors which influenced the community in a higher degree were the salinity and oxygen. The bacteria belonging to Bacteroidetes and Proteobacteria were positively influenced by salinity. Nevertheless, bacteria related to Alpha- and Betaproteobacteria classes and phylum Firmicutes showed a positive correlation with oxygen and pH but negative with salinity. The phylumCyanobacteria were less influenced by the environmental variables. The bacterial community composition of Rambla Salada was also studied by dilution-to-extinction technique. Using this method, 354 microorganisms were isolated. The 16S sequences of 61 isolates showed that the diversity was very different to those obtained by DGGE and with those obtained previously by using classic culture techniques. The taxa identified by dilution-to-extinction were Proteobacteria (81.92%), Firmicutes (11.30%), Actinobacteria (4.52%), and Bacteroidetes (2.26%) phyla with Gammaproteobacteria as predominant class (65.7%). The main genera were: Marinobacter (38.85%), Halomonas (20.2%), and Bacillus (11.2%). Nine of the 61 identified bacteria showed less than 97% sequence identity with validly described species and may well represent new taxa. The number of bacteria in different samples, locations, and seasons were calculated by CARD-FISH, ranging from 54.3 to 78.9% of the total prokaryotic population. Inconclusion, the dilution-to-extinction technique could be a complementary method to classical culture based method, but neither gets to cultivate the major taxa detected by DGGE. The bacterial community was influenced significantly by the physico-chemical parameters (specially the salinity and oxygen), the location and the season of sampling.This research was supported by grants from the Dirección General de Investigación Científica y Técnica (CGL2005-05947; CGL2008-02399; CGL2011-25748), Ministerio de Economía y Competitividad and from the Plan Andaluz de Invesigacion (P07-CVI-03150; CVI06226), Spain

Biological Control of Verticillium Wilt on Olive Trees by the Salt-Tolerant Strain Bacillus velezensis XT1

Verticillium wilt, caused by the pathogen Verticillium dahliae, is extremely devastating to olive trees (Olea europea). Currently, no successful control measure is available against it. The objective of this work was to evaluate the antifungal activity of Bacillus velezensis XT1, a well-characterized salt-tolerant biocontrol strain, against the highly virulent defoliating V. dahliae V024. In vitro, strain XT1 showed to reduce fungal mycelium from 34 to 100%, depending on if the assay was conducted with the supernatant, volatile compounds, lipopeptides or whole bacterial culture. In preventive treatments, when applied directly on young olive trees, it reduced Verticillium incidence rate and percentage of severity by 54 and ~80%, respectively. It increased polyphenol oxidase (PPO) activity by 395%, indicating an enhancement of disease resistance in plant tissues, and it decreased by 20.2% the number of fungal microsclerotia in soil. In adult infected trees, palliative inoculation of strain XT1 in the soil resulted in a reduction in Verticillium symptom severity by ~63%. Strain XT1 is biosafe, stable in soil and able to colonize olive roots endophytically. All the traits described above make B. velezensis XT1 a promising alternative to be used in agriculture for the management of Verticillium wilt.Xtrem Biotech S.LEuropean Union SME Instrument XTOnE-774657Spanish Ministry of Economy and Competitiveness SNEO-20161037ESpanish Government CGL2011-25748Spanish Ministry of Industry, Trade and Tourism (project VertiSOLUTION)Ramon y Cajal program from the Ministry of Economy and Competitiveness RYC-2014-15532CEI-BioTic grant of the BioTic Campus of International Excellence CAEP2-46Ministry of Economy and Competitiveness DI-14-0686

Peribacillus castrilensis sp. nov.: A Plant-Growth-Promoting and Biocontrol Species Isolated From a River Otter in Castril, Granada, Southern Spain

A strictly aerobic, chemoheterotrophic, endospore-forming, Gram-positive, rod-shaped bacterial strain N3T was isolated from the feces of a river otter in Castril (Granada, southern Spain). It is halotolerant, motile, and catalase-, oxidase-, ACC deaminase-, and C4- and C8-lipase-positive. It promotes tomato plant growth and can reduce virulence in Erwinia amylovora CECT 222T and Dickeya solani LMG 25993T through interference in their quorum-sensing systems, although other antagonistic mechanisms could also occur. A phylogenetic analysis of the 16S rRNA gene sequence as well as the phenotypic and phylogenomic analyses indicated that the strain N3T is a novel species of the genus Peribacillus, with the highest 16S rRNA sequence similar to that of Bacillus frigoritolerans DSM 8801T (99.93%) and Peribacillus simplex DSM 1321T (99.80%). Genomic digital DNA–DNA hybridization (dDDH) between the strain N3T and Bacillus frigoritolerans DSM 8801T and Peribacillus simplex was 12.8 and 69.1%, respectively, and the average nucleotide identity (ANIb) of strain N3T and Bacillus frigoritolerans DSM 8801T and Peribacillus simplex was 67.84 and 93.21%, respectively. The genomic G + C content was 40.3 mol%. Its main cellular fatty acids were anteiso-C15 : 0 and iso-C15 : 0. Using 16S rRNA phylogenetic and in silico phylogenomic analyses, together with the chemotaxonomic and phenotypic data, we demonstrated that the type strain N3T (=CECT 30509T = LMG 32505T) is a novel species of the genus Peribacillus and the name Peribacillus castrilensis sp. nov. is proposed.Spanish Ministerio de Educación y Ciencia (AGL-2015-68806-R)Spanish Ministry of the Economy and Competitiveness (PID2019-106704RB-100/AEI/10.13039/501100011033)Plan Andaluz de Investigación (Research Group BIO 188)University of Granada Programme (Empleo Garantía Juvenil)Spanish Ministerio de Educación, Cultura y Deporte (FPU15/01717

Potential of the quorum-quenching and plant-growth promoting halotolerant Bacillus toyonensis AA1EC1 as biocontrol agent

The use of fertilizers and pesticides to control plant diseases is widespread in intensive farming causing adverse effects together with the development of antimicrobial resistance pathogens. As the virulence of many Gram-negative phytopathogens is controlled by N-acyl- homoserine lactones (AHLs), the enzymatic disruption of this type of quorum-sensing (QS) signal molecules, mechanism known as quorum quenching (QQ), has been proposed as a promising alternative antivirulence therapy. In this study, a novel strain of Bacillus toyonensis isolated from the halophyte plant Arthrocaulon sp. exhibited numerous traits associated with plant growth promotion (PGP) and degraded a broad range of AHLs. Three lactonases and an acylase enzymes were identified in the bacterial genome and verified in vitro. The AHL-degrading activity of strain AA1EC1 significantly attenuated the virulence of relevant phytopathogens causing reduction of soft rot symptoms on potato and carrots. In vivo assays showed that strain AA1EC1 significantly increased plant length, stem width, root and aerial dry weights and total weight of tomato and protected plants against Pseudomonas syringae pv. tomato. To our knowledge, this is the first report to demonstrate PGP and QQ activities in the species B. toyonensis that make this strain as a promising phytostimulant and biocontrol agent.Spanish Ministry for Science and Innovation/Agencia Estatal de Investigación 10.13039/501100011033ESF Investing in your future (Grant: RYC2019- 026481-I)Spanish Ministry for Science and Innovation/Agencia Estatal de Investigación 10.13039/501100011033 (Grant: PID2019-106704RB-100)Consejería de Universidad, Investigación e Innovación de la Junta de AndalucíaERDF A way of making Europe (Grant: B-AGR-222-UGR20

Microorganisms isolated from Saharan dust intrusions in the Canary Islands and processes of mineral atmospherogenesis

Global warming due to climate change has increased the frequency of sand and dust storms that affect air quality and ecosystems in general, contributing to air pollution. The Sahara Desert is the most potent emitter of atmospheric dust. The atmosphere is an extreme environment and microorganisms living in the troposphere are exposed to greater ultraviolet radiation, desiccation, low temperatures and nutrient deprivation than in other habitats. The Iberian Peninsula, and specifically the Canary Islands—due to its strategic location—is one of the regions that receive more Saharan dust particles annually, increasing year after year, although culturable microorganisms had previously never been described. In the present work, dust samples were collected from three calima events in the Canary Islands between 2021 and 2022. The sizes, mineralogical compositions and chemical compositions of dust particles were determined by laser diffraction, X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS), respectively. Particle morphology and biological features were also studied by scanning electron microscopy (SEM-EDX) and confocal laser scanning microscopy (CLSM). The mineral–bacteria interactions were described from microscopic observations, which revealed the presence of iberulites and small neoformed kaolinite crystals in association with bacteria. This article defines the term “mineral atmospherogenesis” and its variant, “mineral bioatmospherogenesis”, through microbial interaction. This is the first described case of kaolinite produced through mineral bioatmospherogenesis. The bacterial growth in atmospheric dust was illustrated in SEM images, constituting a novel finding. Twenty-three culturable microorganisms were isolated and identified by 16S rRNA sequencing. Members of the phyla Pseudomonadota, Bacillota and Actinomycetota have been found. Some of these microorganisms, such as Peribacillus frigoritolerans, have Plant Growth-Promoting Rhizobacteria (PGPR) properties. Potential human pathogenic bacteria such as Acinetobacter lwoffii were also found. The presence of desert dust and iberulites in the Canary Islands, together with transported biological components such as bacteria, could have a significant impact on the ecosystem and human health

A new teaching strategy to teach microbiology through its history

Ante el nuevo reto de la implantación del Espacio Europeo de Educación Superior los métodos de estudio deben adaptarse y servirse de herramientas innovadoras. Una buena estrategia para comprender una ciencia, su metodología y objetivos, es conocer su historia. Esta estrategia se hace especialmente útil en el caso de la Microbiología por ser una disciplina joven, que se ha cimentado precisamente en la metodología que sus creadores han ido elaborando. El objetivo de este trabajo ha sido crear una herramienta de utilidad en el aprendizaje de la Microbiología. Para ello hemos tomado como punto de apoyo su historia, es decir, explicar la Microbiología mediante la descripción de los descubrimientos y hechos que contribuyeron al desarrollo de esta ciencia. Para ello se ha creado una página web con dos aplicaciones independientes que operan sobre una base de datos común. 1. La primera de carácter público para los alumnos aunque con posibilidad de acceso restringido; se pueden consultar los contenidos sin posibilidad de alterarlos. 2. La otra aplicación es privada, los profesores pueden administrar los contenidos, y hacer uso de diferentes herramientas que facilitan la gestión de los mismos.In the light of the challenge presented by the European Higher Education Area, study strategies will have to adapt themselves and take advantage of innovative tools provided by modern information technology. One good way of understanding a science, its methodology and objectives is by taking an interest in its history. This approach is especially useful with regard to Microbiology, which is a fairly young discipline founded upon and made cohesive by the methods devised and constantly elaborated on by its designers. The aim of this work has been to offer guidance towards learning Microbiology. To this end we have chosen as our starting point, and indeed one of the keystones of our approach, the history of the subject itself; that is to say, an explanation of Microbiology via a description of the discoveries and milestones that have contributed to the development of this science. Thus we have set up a web page with two separate applications operating on one common database. 1. The first is openly available to the students, although allowing the possibility of restricted access. Its contents may be consulted but not altered. 2. The second is accessible only by lecturers, who may administer its contents and resort to different tools to facilitate the management of the information available on the site

Origin of pharmaceutical care in Spain: the Congress of Pharmaceutical Sciences de Alcalá de Henares, 1995

Introducción La diversidad microbiana es una fuente importante de productos de interés biotecnológico, tales como antibióticos, enzimas o Polímeros11,12. Actualmente la biodiversidad está viéndose seriamente afectada por la contaminación y por la intervención del hombre en la naturaleza. La pérdida de microorganismos no solo alterará los ecosistemas sino que supondrá la desaparición de productos de interés biotecnológico. La biodiversidad microbiana permanece aún inexplorada porque con los métodos clásicos de cultivo microbiano sólo aislamos en el laboratorio entre el 0,1 y el 10% de los microorganismos presentes en determinado ecosistema. En cambio la aplicación de herramientas moleculares permite estudiar la diversidad, estructura y la dinámica de comunidades microbianas de diferentes y complejos ambientes, así como detectar la presencia de microorganismos con interés para la industria farmacéutica y la agricultura, entre otras áreas. Objetivo Analizar y estudiar la diversidad microbiana mediante técnicas moleculares, tanto en ambientes libres de contaminación como es el Parque natural Rambla Salada (Murcia), como en otros hábitats contaminados por las actividades humanas (suelos agrícolas de Motril, Granada). Metodología El estudio de la diversidad microbiana requirió la puesta a punto de una estrategia molecular basada en la amplificación por PCR del gen ribosomal 16S rRNA, a partir del DNA total extraído directamente de las muestras. Posteriormente, se desarrolló la electroforesis en gel con gradiente desnaturalizante (DGGE)1 para la separación de los fragmentos del gen ribosomal 16S ya amplificados, y finalmente se procedió a la secuenciación y comparación de las secuencias obtenidas a partir de los patrones del DGGE con las existentes en las bases de datos. Por otro lado se desarrolló la técnica de hibridación in situ7 empleando sondas fluorescentes; estas últimas fueron diseñadas para detectar la presencia de microorganismos pertenecientes a los dominios de procariotas Bacteria y Arquea. Discusión y conclusión Mediante las herramientas moleculares hemos detectado la presencia de arqueas y bacterias no halófilas, halófilas y halotolerantes tanto en todas las zonas analizadas de Rambla Salada como algunas muestras de los suelos agrícolas de Motril. La diversidad microbiana de los suelos agrícolas de Motril es inferior a la de Rambla Salada. Se ha puesto de manifiesto la existencia de una gran diversidad microbiana en Rambla Salada compuesta fundamentalmente por procariotas pertenecientes a los phila Bacteroidetes, Cianobacteria, Proteobacteria, Firmicutes y Actinobacteria cuyos miembros no se aíslan en el laboratorio mediante las técnicas de cultivo empleadas hasta el momento. La presencia de una mayor diversidad microbiana en Rambla Salada indica que los hábitats hipersalinos son una fuente potencial de productos de interés farmacéutico.Microbial diversity is an important source of products that have potential biotechnological applications, such as antibiotics, enzymes or polymers [1, 2]. The biodiversity is seriously affected by pollution and human impact on natural environments. In this sense, reduction of biodiversity not only alters the ecosystems but also will entail the disappearance of products having biotechnological interest. Microbial biodiversity is not enough known because we have only be able so far to culture between 0.1 and 10% of the microorganisms that exist in nature. Nevertheless, molecular ecology techniques represent an opportunity to study the diversity, structure and dynamics of these uncultured microorganisms, both in diverse and complex environments. They also are useful tools to detect Microorganisms that are of interest to pharmaceutical industry and agriculture, among to other areas. Therefore, the main objective of this work has been to analyse the microbial diversity in different environments using molecular methods. The habitats studied were an hypersaline soil located at Rambla Salada (Murcia), and some agricultural soils from Motril (Granada). We used PCR/DGGE to investigate the microbial diversity; this method is based on the amplification of partial 16S RNAr gene using the total DNA extracted directly from the sample. Subsequently denaturant gradient gel electrophoresis (DGGE) [3] is developed to separate the amplified fragments of the 16S RNAr gene. Finally the sequences obtained from the DGGE patterns are compared to those available at the database. On the other hand, we used fluorescence in situ hybridization or FISH [4] to detect and quantify microorganisms belonging to the Domain Bacteria and Archaea. According to our results, non halophilic, halophilic, and halotolerant Archaea and Bacteria were present in all the areas analyzed at Rambla Salada, as well as in some samples of the agricultural soils in Motril. Microbial diversity found in the agricultural soils in Motril was lower than in Rambla Salada. Microbial community at Rambla Salada was composed of Prokaryotes belonging to the phyla Bacteroidetes, Cyanobacteria, Proteobacteria, Firmicutes and Actinobacteria, which cannot not so far isolated using methods based on traditional culture techniques

Enhanced NSAIDs Solubility in Drug–Drug Formulations with Ciprofloxacin

Drug–drug salts are a kind of pharmaceutical multicomponent solid in which the two co-existing components are active pharmaceutical ingredients (APIs) in their ionized forms. This novel approach has attracted great interest in the pharmaceutical industry since it not only allows concomitant formulations but also has proved potential to improve the pharmacokinetics of the involved APIs. This is especially interesting for those APIs that have relevant dose-dependent secondary effects, such as non-steroidal anti-inflammatory drugs (NSAIDs). In this work, six multidrug salts involving six different NSAIDs and the antibiotic ciprofloxacin are reported. The novel solids were synthesized using mechanochemical methods and comprehensively characterized in the solid state. Moreover, solubility and stability studies, as well as bacterial inhibition assays, were performed. Our results suggest that our drug–drug formulations enhanced the solubility of NSAIDs without affecting the antibiotic efficacy.FPI grant B-FQM-478-UGR20 AGL-2015-68806-RSpanish Agencia Estatal de Investigacion of the Ministerio de Ciencia e Innovacio

Halomonas maura TK26 (CECT 5720) un mutante que sintetiza con alto rendimiento el exopolisacárido maurano26

Número de solicitud: 200202041Halamonas maura TK26(CECT 5720) obtenido mediante mutación biológica de la cepa salvaje de halomonas maura S30 con el transposón mini-Tn5. El cultivo de esta bacteria permite obtener elevadas cantidades de polisacárido maurano26 de elevado poder viscosizante y emulgente, y capaz de captar metales pesados y otros cationes. El polisacárido se obtiene con un rendimiento de unos 5 g/l de medio de cultivo. El polímero presenta una masa molecular de 5.3 x 10(elevado a la sexta) Da. Esta compuesto fundamentalmente por glucosa, manosa, galactosa; además posee un elevado porcentaje de resto de acetilo, sulfato y ácidos urónicos. Los cultivos de microorganismo no son viscosos y la presencia de sales en los mismos disminuye las contaminaciones. Dadas sus propiedades funcionales, este polímero tiene un elevado interés para la industria alimenticia, farmacéutica, y cosmética por su actividad viscosizante y estabilizadora de emulsiones y procesos de biodetoxificación por su capacidad para captar cationes

ZIF-8 and Its Magnetic Functionalization as Vehicle for the Transport and Release of Ciprofloxacin

The use of nanomaterials for the controlled release of drugs aims to enhance their effectiveness, especially when poorly soluble in water, and achieve their rapid, localized, and effective administration. The present study focuses on the use of a Zeolitic Imidazolate Framework-8 (ZIF-8) as vehicle for the transport and controlled release of the antibiotic ciprofloxacin (CIP) as model due to its favorable physicochemical characteristics. The objective is to synthesize the ZIF-8 material loaded with CIP through encapsulation for subsequent release of the drug in neutral and acid physiological media. In addition, functionalization of the CIP/ZIF compound with magnetic nanoparticles (NP) was sought to increase its traceability through the possible use of magnetic fields. Characterizations by XRD, FT-IR, SEM-EDX, and TGA showed a satisfactory synthesis of both pure ZIF-8 and ciprofloxacin-loaded ZIF-8, with high crystallinity and thermal stability. The release profiles showed an abrupt initial release that stabilized over time. A much higher release (20–80% greater) was obtained in acid versus neutral pH in all cases, attributable to the collapse of the ZIF-8 structure in acid media. In addition, functionalization of the material with iron NPs did not affect the behavior of the system during drug release. Antimicrobial activity tests against E. coli and S. aureus showed that ZIF-8 per se exerts antimicrobial activity, while the compounds CIP/ZIF and magnetic CIP/ZIF increased the antimicrobial capacity of pure CIP by 10–20%. The ZIF-8 system has high potential as a drug carrier and release agent for the treatment of diseases, especially those that cause acidification of the cellular environment, achieving a rapid, localized, and targeted action with the possibility of achieving traceability of the system after its magnetic functionalization.P18-RT-419