Browsing by Subject "Bioaerosol"
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- PublicationOpen AccessA new putative Caulimoviridae genus discovered through air metagenomics(American Society for Microbiology, 2018-10-11) Rastrojo, Alberto; Núñez, Andrés; Moreno, Diego A.; Alcamí, Antonio; Genética y MicrobiologíaMembers of the Caulimoviridae family are important plant pathogens. These circular double-stranded DNA viruses may integrate into the host genome, although this integration is not required for the viral replication cycle. Here, we describe three complete genomes belonging to a new putative Caulimoviridae genus discovered through air metagenomics.
- PublicationEmbargoArtemisia pollen is the main vector for airborne endotoxin(Elsevier, 2018-08-09) Oteros, José; Bartusel, Elke; Alessandrini, Francesca; Núñez, Andrés; Moreno, Diego A.; Behrendt, Heidrun; Schmidt-Weber, Carsten; Traidl-Hoffmann, Claudia; Buters, Jeroen; Genética y MicrobiologíaBackground: Endotoxin (LPS) released from gram-negative bacteria causes strong immunologic and inflammatory effects and, when airborne, can contribute to respiratory conditions, such as allergic asthma. Objectives: We sought to identify the source of airborne endotoxin and the effect of this endotoxin on allergic sensitization. Methods: We determined LPS levels in outdoor air on a daily basis for 4 consecutive years in Munich (Germany) and Davos (Switzerland). Air was sampled as particulate matter (PM) greater than 10 μm (PM > 10) and PM between 2.5 and 10 μm. LPS levels were determined by using the recombinant Factor C assay. Results: More than 60% of the annual endotoxin exposure was detected in the PM > 10 fraction, showing that bacteria do not aerosolize as independent units or aggregates but adhered to large particles. In Munich 70% of annual exposure was detected between June 12th and August 28th. Multivariate modeling showed that endotoxin levels could be explained by phenological parameters (ie, plant growth). Indeed, days with high airborne endotoxin levels correlated well with the amount of Artemisia pollen in the air. Pollen collected from plants across Europe (100 locations) showed that the highest levels of endotoxin were detected on Artemisia vulgaris (mugwort) pollen, with little on other pollen. Microbiome analysis showed that LPS concentrations on mugwort pollen were related to the presence of Pseudomonas species and Pantoea species communities. In a mouse model of allergic disease, the presence of LPS on mugwort pollen was needed for allergic sensitization. Conclusions: The majority of airborne endotoxin stems from bacteria dispersed with pollen of only one plant: mugwort. This LPS was essential for inducing inflammation of the lung and allergic sensitization.
- PublicationOpen AccessAssessment and statistical modelling of airborne microorganisms in Madrid(Elsevier, 2020-11-21) Cordero, José María; Núñez, Andrés; García, Ana M.; Borge, Rafael; Genética y MicrobiologíaThe limited evidence available suggests that the interaction between chemical pollutants and biological particles may intensify respiratory diseases caused by air pollution in urban areas. Unlike air pollutants, which are routinely measured, records of biotic component are scarce. While pollen concentrations are daily surveyed in most cities, data related to airborne bacteria or fungi are not usually available. This work presents the first effort to understand atmospheric pollution integrating both biotic and abiotic agents, trying to identify relationships among the Proteobacteria, Actinobacteria and Ascomycota phyla with palynological, meteorological and air quality variables using all biological historical records available in the Madrid Greater Region. The tools employed involve statistical hypothesis contrast tests such as Kruskal-Wallis and machine learning algorithms. A cluster analysis was performed to analyse which abiotic variables were able to separate the biotic variables into groups. Significant relationships were found for temperature and relative humidity. In addition, the relative abundance of the biological phyla studied was affected by PM10 and O3 ambient concentration. Preliminary Generalized Additive Models (GAMs) to predict the biotic relative abundances based on these atmospheric variables were developed. The results (r = 0.70) were acceptable taking into account the scarcity of the available data. These models can be used as an indication of the biotic composition when no measurements are available. They are also a good starting point to continue working in the development of more accurate models and to investigate causal relationships.
- PublicationEmbargoComprehensive analysis of different adhesives in aerobiological sampling using optical microscopy and high-throughput DNA sequencing(Elsevier, 2019-04-05) Rojo, Jesús; Núñez, Andrés; Lara, Beatriz; Sánchez-Parra, Beatriz; Moreno, Diego A.; Pérez-Badía, Rosa; Genética y MicrobiologíaThe standardization and unification of the procedures to analyze and quantify the airborne pollen concentrations are very important topics. In this work, the effectiveness of the two most used adhesives in aerobiological sampling, silicone prepared with cyclohexane solvent (Silicone) and petroleum jelly (Vaseline), was compared under outdoor conditions. This comparison was carried out using the traditional method based on the identification and quantification by optical microscopy (OM) of the airborne pollen and the novel methodology by high-throughput sequencing analysis (HTS). Globally, the results from both methods of analysis (OM and HTS) showed a good agreement between the two adhesives tested regarding the abundance of the main pollen types present in the samples: Cupressaceae, Olea, Poaceae, Platanus, Quercus. We concluded that the results from both adhesives are comparable data. Furthermore, the comparisons between methodologies, OM vs. HTS, showed that both techniques can accurately identify the most abundant pollen types in the atmosphere for the studied periods, with a good agreement of their relative abundances especially when the airborne pollen diversity is low but showing some divergences as the number of pollen types increases.
- PublicationEmbargoDistribution of airborne pollen, fungi and bacteria at four altitudes using high-throughput DNA sequencing(Elsevier, 2020-10-11) Sánchez-Parra, Beatriz; Núñez, Andrés; García, Ana M.; Campoy, Pascual; Moreno, Diego A.; Genética y MicrobiologíaThe diversity and composition of bioaerosols in the atmosphere have been studied in the recent years in multiple places and under very different meteorological conditions. Pollen grains, fungal propagules and bacteria are the most studied airborne biological particles, usually individually, and correlated to environmental factors such as temperature, wind speed, rain, air pollution, etc. to understand their influence. However, the effect of the height on the distribution and composition of these bioaerosols has not been properly described so far. In this research, we analyzed concurrently these three biological particles at four different heights (1.5 m, 130 m, 500 m and 1000 m) by high-throughput DNA sequencing. We observed that the atmosphere contains a homogeneous distribution and composition of biological particles regardless of the altitude tested. Only a minority of the taxa is differently distributed. Therefore, we here deduce that the altitude does not drastically influence the airborne bioparticles distribution and composition under similar meteorological conditions. We also studied the co-occurrence between different bioaerosols, finding new potential relationships.
- PublicationOpen AccessMonitoring of airborne biological particles in outdoor atmosphere. Part 1: Importance, variability and ratios(Hemeroteca Científica Catalana [University Publisher], 2016-03-10) Núñez, Andrés; Amo de Paz, Guillermo; Rastrojo, Alberto; García, Ana M.; Alcamí, Antonio; Gutiérrez-Bustillo, A. Montserrat; Moreno, Diego A.; Genética y MicrobiologíaThe first part of this review ("Monitoring of airborne biological particles in outdoor atmosphere. Part 1: Importance, variability and ratios") describes the current knowledge on the major biological particles present in the air regarding their global distribution, concentrations, ratios and influence of meteorological factors in an attempt to provide a framework for monitoring their biodiversity and variability in such a singular environment as the atmosphere. Viruses, bacteria, fungi, pollen and fragments thereof are the most abundant microscopic biological particles in the air outdoors. Some of them can cause allergy and severe diseases in humans, other animals and plants, with the subsequent economic impact. Despite the harsh conditions, they can be found from land and sea surfaces to beyond the troposphere and have been proposed to play a role also in weather conditions and climate change by acting as nucleation particles and inducing water vapour condensation. In regards to their global distribution, marine environments act mostly as a source for bacteria while continents additionally provide fungal and pollen elements. Within terrestrial environments, their abundances and diversity seem to be influenced by the land-use type (rural, urban, coastal) and their particularities. Temporal variability has been observed for all these organisms, mostly triggered by global changes in temperature, relative humidity, et cetera. Local fluctuations in meteorological factors may also result in pronounced changes in the airbiota. Although biological particles can be transported several hundreds of meters from the original source, and even intercontinentally, the time and final distance travelled are strongly influenced by factors such as wind speed and direction.
- PublicationOpen AccessMonitoring of airborne biological particles in outdoor atmosphere. Part 2: metagenomics applied to urban environments(Institut d'Estudis Catalans, 2016-11-15) Núñez, Andrés; Amo de Paz, Guillermo; Rastrojo, Alberto; García, Ana M.; Alcamí, Antonio; Gutiérrez-Bustillo, A. Montserrat; Moreno, Diego A.; Genética y MicrobiologíaThe air we breathe contains microscopic biological particles such as viruses, bacteria, fungi and pollen, some of them with relevant clinic importance. These organisms and/or their propagules have been traditionally studied by different disciplines and diverse methodologies like culture and microscopy. These techniques require time, expertise and also have some important biases. As a consequence, our knowledge on the total diversity and the relationships between the different biological entities present in the air is far from being complete. Currently, metagenomics and next-generation sequencing (NGS) may resolve this shortage of information and have been recently applied to metropolitan areas. Although the procedures and methods are not totally standardized yet, the first studies from urban air samples confirm the previous results obtained by culture and microscopy regarding abundance and variation of these biological particles. However, DNA-sequence analyses call into question some preceding ideas and also provide new interesting insights into diversity and their spatial distribution inside the cities. Here, we review the procedures, results and perspectives of the recent works that apply NGS to study the main biological particles present in the air of urban environments.
- PublicationEmbargoPreventing legionellosis outbreaks by a quick detection of airborne Legionella pneumophila(Elsevier, 2019-01-15) Sánchez-Parra, Beatriz; Núñez, Andrés; Moreno, Diego A.; Genética y MicrobiologíaLegionellosis is a severe pneumonic infection caused by inhaling bacteria of the genus Legionella. Most cases reported in the USA and Europe are associated with the species Legionella pneumophila. This Gram-negative bacterium can survive within a wide spectrum of temperatures, and be transmitted via aerosols from multiple aquatic sources: fountains, thermal spas and other water systems. Although the PCR is one of the most popular methods to verify its presence in environmental or clinical samples, the direct application of this technique to ambient air samples is unusual because of the scarce material in the specimens. Here, we have developed a two-PCR assay, carried out over the V3 and V5 hypervariable regions of the 16S rRNA gene, to detect specifically the pathogenic bacteria Legionella pneumophila in outdoor air samples with low concentration of DNA. The application of this protocol does not require culture and retrieves quick results to activate the corresponding public alerts to prevent legionellosis outbreaks.
- PublicationOpen AccessSaharan dust storms affecting the center of the Iberian Peninsula: effect on the urban aerobiome(Elsevier, 2024-04-16) Núñez, Andrés; Moreno, Diego A.; García, Ana M.; Genética y MicrobiologíaDust storms are known to be atmospheric phenomena that transport mineral dust but also airborne biological particles (bioaerosols) from desert areas to distant regions. These bioaerosols can influence atmospheric processes and they have the potential of changing the composition of the local aerobiome in urban areas, which, in recent years, have been associated with allergies and the exacerbation of respiratory syndromes. Here, we studied four dust events initiated in the Sahara Desert affecting the center of the Iberian Peninsula. The biological particles before and during the phenomena were analyzed by high-throughput DNA sequencing. The global composition of bioaerosols showed a marked seasonality. The relative abundances of the most predominant groups of bacteria and fungi were not significantly altered compared to the days prior the corresponding event. Nonetheless, we detected specific bacterial and fungal taxa associated with these events, whose composition and abundance were also related to the period of the year. Although a variety of plant and animal pathogens were identified both before and throughout the days influenced by dust storms, some were only detected during the latter, suggesting a long-range transport for these species. This work highlights the importance of analyzing the storm dust events separately, especially when they occur at different seasons, and the particular effect on an urban environment in the Iberian Peninsula as a model case, providing some recommendations for future studies.
- PublicationOpen AccessSeasonal changes dominate long-term variability of the urban air microbiome across space and time(Elsevier, 2021-02-09) Núñez, Andrés; García, Ana M.; Moreno, Diego A.; Guantes, Raúl; Genética y MicrobiologíaCompared to soil or aquatic ecosystems, the atmosphere is still an underexplored environment for microbial diversity. In this study, we surveyed the composition, variability and sources of microbes (bacteria and fungi) in the near surface atmosphere of a highly populated area, spanning ~ 4,000 Km2 around the city center of Madrid (Spain), in different seasonal periods along two years. We found a core of abundant bacterial genera robust across space and time, most of soil origin, while fungi were more sensitive to environmental conditions. Microbial communities showed clear seasonal patterns driven by variability of environmental factors, mainly temperature and accumulated rain, while local sources played a minor role. We also identified taxa in both groups characteristic of seasonal periods, but not of specific sampling sites or plant coverage. The present study suggests that the near surface atmosphere of urban environments contains an ecosystem stable across relatively large spatial and temporal scales, with a rather homogenous composition, modulated by climatic variations. As such, it contributes to our understanding of the long-term changes associated to the human exposome in the air of highly populated areas.
- PublicationEmbargoTemporal patterns of variability for prokaryotic and eukaryotic diversity in the urban air of Madrid (Spain)(Elsevier, 2019-09-10) Núñez, Andrés; Amo de Paz, Guillermo; Rastrojo, Alberto; Ferencova, Zuzana; Gutiérrez-Bustillo, A. Montserrat; Alcamí, Antonio; Moreno, Diego A.; Guantes, Raúl; Genética y MicrobiologíaAlthough many microorganisms are ubiquitously present in the air, airborne microbial communities have been much less characterized than those in soil or aquatic environments. Besides its ecological importance, detection and monitoring of the wide diversity of these aerosolized microorganisms (bacteria, viruses, fungi and pollen) is relevant for understanding allergy and disease outbreaks, especially in highly populated cities. In this study, we describe the simultaneous biodiversity of bacteria, fungi and plants present in the urban atmosphere of Madrid (Spain) along different seasonal periods, using DNA sequencing. Sampling in two different locations (downtown and peri-urban) we found that changes in the composition of each community are mainly driven by environmental factors, rather than by the features of the specific sampling microenvironments. While pollen particles are dominated by a few taxa characteristic of each season, bacteria and fungi show a high diversity but stable core communities along the year. The prokaryotic core is governed by soil and leaf surface bacteria, with predominance of Actinobacteria (Frankiales and Micrococcales) and Alphaproteobacteria (Sphingomonadales, Rhodobacterales, Rhizobiales and Acetobacterales). Fungal diversity is characterized by the steady presence of members of Capnodiales and Pleosporales. Pathogenic bacterial and fungal taxa were also detected across the year. We also correlated the airborne biodiversity with environmental variables. Air temperature has a strong influence on the community composition of bacteria, while pollen and fungi seasonal variations are mainly correlated with precipitation. Our results contribute to the characterization of airborne prokaryotic and eukaryotic communities in urban areas and show the suitability of this method for biosurveillance strategies.
- PublicationOpen AccessThe differential vertical distribution of the airborne biological particles reveals an atmospheric reservoir of microbial pathogens and aeroallergens(Springer, 2020-03-27) Núñez, Andrés; Moreno, Diego A.; Genética y MicrobiologíaThe most abundant biological particles present in the air are bacteria, fungal propagules and pollen grains. Many of them are proved allergens or even responsible for airborne infectious diseases, which supports the increase of studies in recent years on their composition, diversity, and factors involved in their variability. However, most studies in urban areas are conducted close to ground level and a factor such as height is rarely taken into account. Thus, the information about how the composition of biological particles changes with this variable is scarce. Here, we examined the differential distribution of bacteria, fungi, and plants at four altitudes (up to ∼ 250 m) in a metropolitan area using high-throughput DNA sequencing. Most taxa were present at all levels (common taxa). However, a transitional layer between 80 and 150 m seemed to affect the scattering of these bioaerosols. Taxa not present at all altitudes (non-common) showed an upward tendency of diversity for bacteria and plants with height, while the opposite trend was observed for fungi. Certain patterns were observed for fungi and specific plant genera, while bacterial taxa showed a more arbitrary distribution and no patterns were found. We detected a wide variety of aeroallergens and potential pathogens at all heights, which summed a substantial portion of the total abundance for fungi and plants. We also identified potential connections between the biological particles based on their abundances across the vertical section.
- PublicationOpen AccessValidation of the hirst-type spore trap for simultaneous monitoring of prokaryotic and eukaryotic biodiversities in urban air samples by next-generation sequencing(American Society for Microbiology, 2017-04-28) Núñez, Andrés; Amo de Paz, Guillermo; Ferencova, Zuzana; Rastrojo, Alberto; Guantes, Raúl; García, Ana M.; Alcamí, Antonio; Gutiérrez-Bustillo, A. Montserrat; Moreno, Diego A.; Genética y MicrobiologíaPollen, fungi, and bacteria are the main microscopic biological entities present in outdoor air, causing allergy symptoms and disease transmission and having a significant role in atmosphere dynamics. Despite their relevance, a method for monitoring simultaneously these biological particles in metropolitan environments has not yet been developed. Here, we assessed the use of the Hirst-type spore trap to characterize the global airborne biota by high-throughput DNA sequencing, selecting regions of the 16S rRNA gene and internal transcribed spacer for the taxonomic assignment. We showed that aerobiological communities are well represented by this approach. The operational taxonomic units (OTUs) of two traps working synchronically compiled >87% of the total relative abundance for bacterial diversity collected in each sampler, >89% for fungi, and >97% for pollen. We found a good correspondence between traditional characterization by microscopy and genetic identification, obtaining more-accurate taxonomic assignments and detecting a greater diversity using the latter. We also demonstrated that DNA sequencing accurately detects differences in biodiversity between samples. We concluded that high-throughput DNA sequencing applied to aerobiological samples obtained with Hirst spore traps provides reliable results and can be easily implemented for monitoring prokaryotic and eukaryotic entities present in the air of urban areas.