Airborne Microbial Communities Research Articles

Microbiomes of air dust collected during the ground-based closed bioregenerative life support experiment "Lunar Palace 365"

BackgroundUnderstanding the dynamics of airborne microbial communities and antibiotic resistance genes (ARGs) in space life support systems is important because potential pathogens and antibiotic resistance pose a health risk to crew that can lead to mission failure. There have been few reports on the distribution patterns of microbiomes and ARGs in space life support systems. In particular, there have been no detailed investigations of microbiomes and/or antibiotic resistance based on molecular methods in long-term confined bioregenerative life support systems (BLSSs). Therefore, in the present study, we collected air dust samples from two crew shifts, different areas, and different time points in the "Lunar Palace 365" experiment. We evaluated microbial diversity, species composition, functional potential, and antibiotic resistance by combining cultivation-independent analyses (amplicon, shot-gun sequencing, and qPCR).ResultsWe found that the bacterial community diversity in the Lunar Palace1 (LP1) system was higher than that in a controlled environment but lower than that in an open environment. Personnel exchange led to significant differences in bacterial community diversity, and source tracking analysis revealed that most bacteria in the air derived from the cabin crew and plants, but no differences in microbial function or antibiotic resistance were observed. Thus, human presence had the strongest effect on the succession of microbial diversity in the BLSSs.ConclusionsOur results highlight that microbial diversity in BLSSs is heavily influenced by changes in crew and is unique from other open and controlled environments. Our findings can be used to help develop safe, enclosed BLSS that meet the requirements of human survival and habitation in outer space. In addition, our results can further enhance our understanding of the indoor air microbial community and effectively maintain a safe working and living environment, including plant growth.

The Response of Airborne Mycobiome to Dust Storms in the Eastern Mediterranean

Airborne microbial communities directly impact the health of humans, animals, plants, and receiving ecosystems. While airborne bacterial and fungal communities have been studied by both cultivation-based methods and metabarcoding surveys targeting specific molecular markers, fewer studies have used shotgun metagenomics to study the airborne mycobiome. We analyzed the diversity and relative abundance of fungi in nine airborne metagenomes collected on clear days (“background”) and during dust storms in the Eastern Mediterranean. The negative correlation between the relative abundance of fungal reads and the concentrations of atmospheric particulate matter having an aerodynamic diameter smaller than 10 μm (PM10) indicate that dust storms lower the proportion of fungi in the airborne microbiome, possibly due to the lower relative abundance of fungi in the dust storm source regions and/or more effective transport of bacteria by the dust. Airborne fungal community composition was altered by the dust storms, particularly those originated from Syria, which was enriched with xerophilic fungi. We reconstructed a high-quality fungal metagenome-assembled genome (MAG) from the order Cladosporiales, which include fungi known to adapt to environmental extremes commonly faced by airborne microbes. The negative correlation between the relative abundance of Cladosporiales MAG and PM10 concentrations indicate that its origin is dominated by local sources and likely includes the indoor environments found in the city.

RNA-Based Analysis Reveals High Diversity of Plant-Associated Active Fungi in the Atmosphere

Fungi are ubiquitous in nature; that is, they are present everywhere on the planet; understanding the active state and functional capacity of airborne microbes associated with health of human, animal, and plant is critical for biosafety management. Here, we firstly and directly proved that there were about 40% active fungi in the air via rRNA amplicon sequencing and imaging flow cytometry simultaneously. Amplicon sequencing analysis showed differences between structures of active and total fungal community; Ascomycota were dominant in the active community, while Basidiomycota have low transcriptional activity across all samples. Notably, plant pathogenic fungi were predominant in the air, and more than 50% were active, including not only several common plant pathogens but also biotrophic fungi (Erysiphe sp. and Microbotryum sp.) and host-specific pathogens, which were generally considered to be inactive after leaving the host. Putative plant pathogens of eight genera were found active across the sampling season, indicating their superior ability to obtain nutrients even in barren nutrient environments. Interestingly, we detected several potentially active unrecorded fungi in China (Diatrype prominens, Septofusidium herbarum, Pseudomicrostroma glucosiphilum, and Uromycladium tepperianum), which suggested that they spread over a long distance by air and may cause diseases under favorable conditions. Our results suggested that maintaining transmission in air is an essential feature of many fungi including plant pathogens regardless of being a biotrophic, hemibiotrophic, or necrotrophic group. Moreover, two potentially active human pathogens and one animal pathogen were captured, which indicated their potential risks. This study provided a new perspective for more comprehensive understanding of airborne fungi, including their multidimensional lifestyle, state, functioning, and potential pathogenic risk. It also laid the foundation for further prediction and management of airborne microbial communities, which would be of interest for public health and agriculture.

Seasonal Variability of the Airborne Eukaryotic Community Structure at a Coastal Site of the Central Mediterranean.

The atmosphere represents an underexplored temporary habitat for airborne microbial communities such as eukaryotes, whose taxonomic structure changes across different locations and/or regions as a function of both survival conditions and sources. A preliminary dataset on the seasonal dependence of the airborne eukaryotic community biodiversity, detected in PM10 samples collected from July 2018 to June 2019 at a coastal site representative of the Central Mediterranean, is provided in this study. Viridiplantae and Fungi were the most abundant eukaryotic kingdoms. Streptophyta was the prevailing Viridiplantae phylum, whilst Ascomycota and Basidiomycota were the prevailing Fungi phyla. Brassica and Panicum were the most abundant Streptophyta genera in winter and summer, respectively, whereas Olea was the most abundant genus in spring and autumn. With regards to Fungi, Botrytis and Colletotrichum were the most abundant Ascomycota genera, reaching the highest abundance in spring and summer, respectively, while Cryptococcus and Ustilago were the most abundant Basidiomycota genera, and reached the highest abundance in winter and spring, respectively. The genus community structure in the PM10 samples varied day-by-day, and mainly along with the seasons. The impact of long-range transported air masses on the same structure was also proven. Nevertheless, rather few genera were significantly correlated with meteorological parameters and PM10 mass concentrations. The PCoA plots and non-parametric Spearman’s rank-order correlation coefficients showed that the strongest correlations generally occurred between parameters reaching high abundances/values in the same season or PM10 sample. Moreover, the screening of potential pathogenic fungi allowed us to detect seven potential pathogenic genera in our PM10 samples. We also found that, with the exception of Panicum and Physcomitrella, all of the most abundant and pervasive identified Streptophyta genera could serve as potential sources of aeroallergens in the studied area.

Characteristics of airborne microbial community in Hefei, eastern China, under a good quality in mid-spring

Characteristics of airborne microbial community in Hefei, eastern China, under a good quality in mid-spring

High-Throughput Sequencing Analysis of Microbial Communities in Summertime Atmospheric Particulate Matter in Hefei City

The composition, physical and chemical properties, sources, and temporal and spatial changes in airborne particulate matter have been extensively investigated in previous studies. However, less is known about bioaerosols, which are mainly composed of bacteria and fungi and constitute up to 25% of the total airborne particulate matter. In this study, we used inductively coupled plasma mass spectrometry and ion chromatography to determine the concentrations of trace elements and water-soluble ions in atmospheric particulates, respectively. These analyses were combined with high-throughput sequencing methods and real-time quantitative polymerase chain reaction to analyze the microbial compositions in PM1.0, PM2.5, and PM10 samples, which were collected from July to September in Hefei City. The results showed that there were no significant differences in the bacterial community diversity across the three size fractions (analysis of variance (ANOVA), P>0.05). The bacterial and fungal community diversities on sunny days were lower than those on rainy days, and the bacterial community diversity in all samples was significantly higher than the fungal community diversity (ANOVA, P<0.01). The predominant bacterial phyla were Proteobacteria (46.19%), Firmicutes (33.42%), Bacteroidetes (10.99%), Cyanobacteria (3.33%), and Actinobacteria (2.11%). Ascomycota (73.23%), Basidiomycota (5.78%), Mortierellomycota (3.41%), and Mucoromycota (0.10%) were the dominant fungal phyla. Our results indicated that soils, plant leaves, and animal feces were the dominant sources of airborne bacterial communities in Hefei City, and the main sources of the fungal communities were plant leaves and soils. The bacterial community was mainly affected by K, Pb, Al, Fe, Mg, Ca, Na+, NO2-, and wind speed, and the main influencing factors of the fungal community were V, Mn, Sr, NO2-, NO3-, Na+, Cl-, the air quality index, and PM10. In addition, nine specific bacteria and fungi that are linked to human health risks were identified, including Acinetobacter, Streptococcus, Enterobacter, Pseudomonas, Delftia, Serratia, Trichoderma, Alternaria, and Aspergillus, which can lead to a wide range of diseases in humans and other organisms. The research results are helpful for revealing the various characteristics of airborne microbial communities, their influencing factors, and their impacts on human health, and are an important reference for subsequent research and the formulation of government policies.

Airborne bacteria and particulate chemistry capture Phytoplankton bloom dynamics in an Arctic fjord

Primary biological aerosol particles and microorganisms are ubiquitous in the atmosphere. Investigations of airborne chemical markers and microbial communities are critical for identifying sources, transport and transformation processes of aerosols. One potential major source of airborne chemical compounds and microbial communities (e.g. L- and D-amino acids, Flavobacteria) could be related to phytoplankton blooms that occur during the spring season in Arctic fjord systems. Here, we conducted a field study in a polar environment to investigate the occurrence in coarse and fine particles of water-soluble compounds (major ions, carboxylic acids and free L- and D-amino acids) and airborne bacterial communities in aerosol samples. The sampling was conducted with a 6 day sampling frequency at the Gruvebadet observatory, close to Ny-Ålesund (Svalbard Islands). Glycine, D-amino acids and C4- organic acids increased during the exponential phase of a marine bloom that occurred in Kongsfjorden and started to drop at the beginning of the main-bloom phase. On the other hand, Polaribacter together with free L-amino acids overlapped with the Chlorophyll a peak and the subsequent decline, and thus might constitute a useful marker for the main-bloom phase.

Urban greenness and plant species are key factors in shaping air microbiomes and reducing airborne pathogens

Urban green space has been implicated in shaping airborne microbes, but there is an only rudimentary understanding of the key factors of urban green space affecting the composition and structures of airborne microbes. Here, we selected 40 urban sites based on stratified random sampling design and investigated the effects of multiple factors including landscapes, plant, soil, and anthropogenic factors on airborne microbial communities, especially bacterial and fungal pathogens. Bacterial and fungal communities in the control area with lower greenness were significantly (P < 0.05) different from those in other areas with a gradient of green space. The relative abundance of bacterial and fungal pathogens significantly (P < 0.05) decreased with increasing greenness. Other than soil thickness, soil type, slope position, and population density, plant species considerably contributed to the shift in the composition and abundance of potential bacterial and fungal pathogens. A significantly (P < 0.05) reduced abundance of bacterial and fungal pathogens was observed in areas with >30% masson pine. Together, these results provide insights into the importance of green space for providing health benefits for city dwellers by reducing pathogens in air, as well as providing support for the inclusion of plant species in the management of urban green space to reduce exposure risk of airborne pathogens.

High time-resolved characterization of airborne microbial community during a typical haze pollution process

Variations of bioaerosol characteristics during the process of haze pollution have rarely been explored. In this study, high time-resolved variations of the community structures of bacteria, fungi, and ammonia-oxidizing microorganisms (AOMs) were assessed during a typical haze pollution process. The impacts of meteorological factors, water-soluble inorganic ions (WSII), and organic dicarboxylic acids (DCA) on the airborne microbial community were systematically evaluated. The results showed that the bacterial community varied greatly during the formation stages of haze pollution, and tended to stabilize with the further development of haze pollution. Nevertheless, variations of the fungal community lasted throughout the whole haze pollution process. Furthermore, Nitrososphaera absolutely dominated the ammonia-oxidizing archaea (AOA) and declined as PM2.5 burst. Network analysis identified relatively weak interactions and co-occurrence patterns between dominant fungal genera. Importantly, dust source ions and PM2.5 acidity exerted the most significant impacts on bacterial and fungal communities. These results identify the high time-resolved variations of airborne microbial communities during the formation and development of haze pollution process, and provide valuable data to better understand the interaction between bioaerosols and haze pollution.

Aerobiological study of bacterial and fungal community composition in the atmosphere of Mexico City throughout an annual cycle.

The atmosphere as a temporary habitat for airborne microbial communities is a valuable topic to explore, and it is through aerobiological studies that the diversity of biological particles and their release, emission, transport, deposition, and impact are assessed. Specific microorganisms are involved in meteorological processes, and phytosanitary and public health concerns. Airborne microbial composition is related to factors such as geographic region and weather conditions. In this study a metagenomic approach was used to determine the composition of bacterial and fungal communities in the air of two different land-use areas (urban area and semi-rural area), during dry and rainy seasons in Mexico City. Air sampling was carried out with a Hirst-type spore trap, collecting the samples simultaneously in both study areas. Forty-two bioaerosol samples were collected, and the DNA obtained was sequenced using Next-Generation Sequencing. The results indicated that the bacterial communities were represented mainly by the phyla Actinobacteria, Proteobacteria, Firmicutes, Bacteroidetes, Cyanobacteria, and the fungal communities by the phyla Ascomycota followed by Basidiomycota. The evident changes in microbial composition were related more to seasonality than to locality, since both UA and SRA showed a high degree of urbanization, despite some differences in land use. Continuous monitoring of atmospheric bioaerosols is essential to determine the influence of meteorological factors on the composition of the aerial microbiota.

Land Use Effects on Airborne Bacterial Communities Are Evident in Both Near-Surface and Higher-Altitude Air

Land use influences the composition of near-surface airborne bacterial communities, and bacteria can be transported through the atmosphere at global scales. The atmosphere mixes vertically, but rigorously assessing whether the effects of land use on atmospheric communities extends to higher altitudes requires examining communities from multiple altitudes collected at a stable location and timeframe. In this study, we collected near-surface (&lt;2 m) and higher-altitude (150 m) air samples from three sites in an agricultural/developed location and a forested/undeveloped location. We used bacterial 16S rRNA amplicon sequencing to compare communities and predict functionality by altitude. Higher-altitude and near-surface communities did not differ in composition within each location. Communities collected above the undeveloped location were equally variable at both altitudes; higher-altitude samples from the developed location predominantly contained Firmicutes and were less variable than near-surface samples. We also compared airborne taxa to those present in soil and snow. Communities from higher-altitude samples above the developed location contained fewer overlapping taxa with soil and snow sources, and overlapping Operational Taxonomic Units (OTUs) among the three sources differed by location. Our results suggest that land use affects the composition of both near-surface and higher-altitude airborne bacterial communities and, therefore, may influence broad bacterial dispersal patterns. This small-scale pilot study provides a framework for simultaneously examining local and regional airborne microbial communities that can be applied to larger studies or studies using different types of samplers.

Variability of the Atmospheric PM10 Microbiome in Three Climatic Regions of France.

Primary Biogenic Organic Aerosols (PBOA) were recently shown to be produced by only a few types of microorganisms, emitted by the surrounding vegetation in the case of a regionally homogeneous field site. This study presents the first comprehensive description of the structure and main sources of airborne microbial communities associated with temporal trends in Sugar Compounds (SC) concentrations of PM10 in 3 sites under a climatic gradient in France. By combining sugar chemistry and DNA Metabarcoding approaches, we intended to identify PM10-associated microbial communities and their main sources at three sampling-sites in France, under different climates, during the summer of 2018. This study accounted also for the interannual variability in summer airborne microbial community structure (bacteria and fungi only) associated with PM10-SC concentrations during a 2 consecutive years’ survey at one site. Our results showed that temporal changes in PM10-SC in the three sites are associated with the abundance of only a few specific taxa of airborne fungi and bacterial. These taxa differ significantly between the 3 climatic regions studied. The microbial communities structure associated with SC concentrations of PM10 during a consecutive 2-year study remained stable in the rural area. Atmospheric concentration levels of PM10-SC species varied significantly between the 3 study sites, but with no clear difference according to site typology (rural vs. urban), suggesting that SC emissions are related to regional rather than local climatic characteristics. The overall microbial beta diversity in PM10 samples is significantly different from that of the main vegetation around the urban sites studied. This indicates that the airborne microorganisms at these urban sites are not solely from the immediate surrounding vegetation, which contrasts with observations at the scale of a regionally homogeneous rural site in 2017. These results improve our understanding of the spatial behavior of tracers of PBOA emission sources, which need to be better characterized to further implement this important mass fraction of Organic Matter (OM) in Chemical Transport models (CTM).

Short-Term Aerial Pollutant Concentrations in a Southwestern China Pig-Fattening House

Concentrations of critical aerial pollutants within animal farms are important to the health of animals and farm staff and can be reduced via manure management, ventilation control, and barn design. This study characterized measurements of ammonia (NH3), total suspended particle (TSP), and airborne microbial communities of a large-scale pig-fattening house, as well as their correlations with environmental variables in Southwestern China. Monitoring was conducted for 15 consecutive days during both August and January, at various locations inside the pig house. The concentrations of NH3 and TSP averaged 3.22 and 0.55 mg m−3, respectively, while the average number of airborne microbial colonies was 3.91 log cfu m−3. The aerial pollutant concentrations displayed significant seasonal differences (p &lt; 0.05). Specifically, concentrations in winter were significantly higher than those in summer (p &lt; 0.05), and the 07:00 measurements were the highest among the three measurement times. The concentrations were significantly correlated with indoor temperature and relative humidity. In summer, TSP concentration was negatively correlated with temperature (correlation coefficient = −0.732), while NH3 concentration was positively correlated with temperature (correlation coefficient = 0.58). In winter, TSP and NH3 concentrations were negatively correlated with relative humidity (correlation coefficients = −0.739 and −0.713, respectively), while the airborne microbial colonies were not correlated with either humidity or temperature in summer or winter. These findings confirm that the aerial pollutant concentrations in a Southwestern China pig-fattening house exhibited significant seasonal and diurnal variations. Air quality can be improved by more precise ventilation control as observed by the correlation of concentrations with ventilation control, indoor temperature, and humidity.

Microbial Flow Within an Air-Phyllosphere-Soil Continuum.

The phyllosphere is populated by numerous microorganisms. Microbes from the wider environment, i.e., air and soil, are considered key contributors to phyllosphere microbial communities, but their contribution is unclear. This study seeks to address this knowledge gap by controlling the movement of microbes along the air-phyllosphere-soil continuum. Customized equipment with dual chambers was constructed that permitted airflow to enter the first chamber while the second chamber recruited filtered microbe-free air from the initial chamber. Allium schoenoprasum (chive) and Sonchus oleraceus (sow thistle) were cultivated in both chambers, and the microbial communities from air, phyllosphere, and soil samples were characterized. Shares of microbial OTUs in the equipment suggested a potential interconnection between the air, phyllosphere, and soil system. Fast expectation-maximization microbial source tracking (FEAST) suggested that soil was the major source of airborne microbial communities. In contrast, the contribution of airborne and soil microbes to phyllosphere microbial communities of either A. schoenoprasum or S. oleraceus was limited. Notably, the soilborne microbes were the only environmental sources to phyllosphere in the second chamber and could affect the composition of phyllosphere microbiota indirectly by air flow. The current study demonstrated the possible sources of phyllosphere microbes by controlling external airborne microbes in a designed microcosm system and provided a potential strategy for recruitment for phyllosphere recruitment.

From the Ground up: Assessing Bacterial Diversity in the Air Space of an Urban Park.

The figure shows an experimental diagram of the vertical stratification sampling stations. There is an illustration of a stand placed on the ground depicting the different levels of sampling. From ...

Influence of pig farming on human Gut Microbiota: role of airborne microbial communities

ABSTRACT It has been hypothesized that both genetics and diet influence the composition of the human cecal microbiota. However, it remains unclear whether and how occupational exposure to microbes impacts the microbial communities in human guts. Using a One Health approach, we visited pig farms (n = 26) and collected stool specimens from pig workers (n = 59), pig barn air samples (n = 19), and rectal swabs from pigs at three different growth stages (n = 144). Stool samples from cattle workers were included as a control group (n = 22). Each sample’s microbiota was characterized using 16S rRNA gene sequencing and the DADA2 pipeline. We obtained a significantly different clustering of the microbial compositions of pig and cattle workers by permutational multivariate analysis of variance (PERMANOVA; P < .001). Workers primarily exposed to pigs had higher relative abundances of Prevotellaceae and less Bacteroidaceae than workers exposed to cattle. We also found that the microbial compositions of pig workers’ stool samples shared extensive fractions with the samples from their pigs. We also identified amplicon sequencing variants (ASVs) in the airborne microbiota which were likely involved in zoonotic transmission events. We hypothesize that ASVs originating from pig feces are aerosolized and, through breathing, get trapped in the pig farm workers’ upper respiratory tract from where they can get swallowed. Consequently, some of the animal associated ASVs are transferred into the gastrointestinal tracts (GITs) which leads to changes in the composition of the human gut microbiota. The importance of this finding for human health must be investigated further.

Taxonomic composition and seasonal dynamics of the air microbiome in West Siberia

Here, we describe taxonomical composition, as well as seasonal and diel dynamics of airborne microbial communities in West Siberia. A total of 78 airborne biomass samples from 39 time intervals were analysed, within a temperature range of 48 °C (26 °C to − 22 °C). We observed a 5–170-fold decrease in DNA yield extracted from the airborne biomass in winter compared to summer, nevertheless, yielding sufficient material for metagenomic analysis. The airborne microbial communities included Actinobacteria and Proteobacteria, Ascomycota and Basidiomycota fungi as major components, as well as some Streptophyta plants. In summer, bacterial and fungal plant pathogens, and wood-rotting saprophytes were predominant. In winter, Ascomycota moulds and cold-related or stress environment bacterial species were enriched, while the fraction of wood-rotting and mushroom-forming Basidiomycota fungi was largely reduced. As recently reported for the tropical climate, the airborne microbial communities performed a diel cycle in summer, however, in winter diel dynamics were not observed.

Compromised Ecosystem Services From Urban Aerial Microbiomes: A Review of Impacts on Human Immune Function

Biodiverse environments contribute to human health through a wide range of ecosystem services: from providing food and medicines to filtering our air and water. Exposure to biodiverse, airborne microbial communities (aerobiomes) contributes to the development of healthy human-immune function. This overlooked but potentially powerful ecosystem service is akin to nature’s provision of traditional medicines and pharmaceutical compounds. But urban environments appear to support less diverse aerobiomes, suppressing this ecosystem service and potentially contributing to urbanassociated diseases through altered immune function. Here, we synthesize the known relationships between aerobiome biodiversity and health and present the experimentally demonstrated mechanisms that connect aerobiome exposure to immune function. We then summarize what is currently known about the effect of urbanization on aerobiomes and identify several important knowledge gaps in this field, including a lack of rigorous, experimental, multi-scale studies demonstrating the mechanistic pathways between urbanization, altered aerobiomes and human health. We offer practical approaches that can close these knowledge gaps and will facilitate the transfer of knowledge and technology between microbiologists, urban ecologists and publichealth practitioners. This synthesis should stimulate interdisciplinary research efforts to advance our understanding of how urbanization is impacting aerobiome ecosystem services, and what that means for human health.

Factors Affecting Levels of Airborne Bacteria in Dairy Farms: A Review.

Simple SummaryThe environmental quality of farms plays an important role in the food safety of the dairy industry because it may influence the microbial communities in milk. The microorganisms present in the different areas of a farm have an influence on this environmental quality, using the air as a vehicle of dissemination. However, the ability of this airborne microbial community to contaminate the milk, like the sources of origin of these microorganisms, has not been well studied in dairy farms until now. This review examines the importance of different factors that affect the environmental quality of dairy farms, and must, therefore, be taken into account when considering the importance of environmental microbiology as a tool in the improvement of the quality of milk and dairy products.This review attempts to reflect the importance of different factors that affect the environmental quality of dairy farms and must, therefore, be taken into account when considering the importance of environmental microbiology as a tool in the improvement of the quality of milk and dairy products. The effect of a factor such as temperature is vital for the dairy farm environment, especially when the temperatures are extreme, because a proper choice of temperature range improves the quality of the air and, thus, animal welfare. Similarly, the appropriate level of relative humidity in the environment should be taken into consideration to avoid the proliferation of microorganisms on the farm. Air quality, well-designed livestock housing, proper hygienic practices on the farm, stocking density, and the materials used in the livestock houses are all important factors in the concentration of microorganisms in the environment, promoting better welfare for the animals. In addition, a ventilation system is required to prevent the pollution of the farm environment. It is demonstrated that proper ventilation reduces the microbial load of the environment of dairy farms, enhancing the quality of the air and, therefore, the wellbeing of the animals. All this information is very useful to establish certain standards on dairy farms to improve the quality of the environment and, thereby, achieve better quality milk and dairy products.

Enumeration and characterization of airborne microbial communities in an outdoor environment of the city of Taj, India

In the present investigation, the microbiological characteristics of the air in the outdoor environment of the city of Taj, i.e., Agra (India) was assessed by determining the concentration and composition of bacteria and fungi using culture technique. The bacterial isolates having the highest prevalence (100%) throughout the year were selected and identified based on morphological, physiological, and biochemical characteristics. The mean concentration of bacteria in PM10 and PM2.5 was 405.1 CFUm-3 and 364.9 CFUm-3, respectively, while the mean concentration of fungi in PM10 and PM2.5 was 136.5 CFUm-3 and 20.3 CFUm-3, respectively. A new technique of BD (Becton Dickinson)BBL Crystal Mind Auto reader was used. The bacteria were identified as Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Micrococcus luteus, Chromobacterium violaceum, Bacillus cereus, Brevibacillus brevis, Enterobacter aerogenes, Pseudomonas aeruginosa, Acinetobacter beijerenckii. Fifteen fungal isolates were identified comprising of Fusarium, Aspergillus, Epicoccum, Penicillium, Cladosporium, Trichoderma. Alternaria, Mucor, Rhizopus, Geotrichum, Helminthosporium, Absidia, Cephalosporium, Botrytis, and Neurospora. Aspergillus and Penicillium have the maximum percentage occurrence of 98% and 95%, respectively. Botrytis recorded a minimum percentage of 22%, followed by Neurospora 20%. The concentration of bacteria and fungi depend upon the meteorological conditions.