scholarly journals Translational activity is uncoupled from nucleic acid content in bacterial cells of the human gut microbiota

2020 ◽  
Author(s):  
Mariia Taguer ◽  
B. Jesse Shapiro ◽  
Corinne F. Maurice
Keyword(s):  
Nucleic Acid ◽  
Gut Microbiota ◽  
Gut Microbiome ◽  
Acid Content ◽  
Membrane Damage ◽  
Bacterial Activity ◽  
Nucleic Acid Content ◽  
Bacterial Cells ◽  
Human Gut ◽  
Human Gut Microbiota

AbstractBackgroundChanges in bacterial diversity in the human gut microbiome, characterized primarily though DNA sequencing methods, have been associated with many different adverse health conditions. However, these changes do not always reflect changes in bacterial activity, and thus how the gut microbiome is implicated in disease is still not often understood. New methods that link together bacterial function to bacterial identity are needed to further explore the role of the gut microbiome in health and disease. We optimized bioorthogonal non-canonical amino acid tagging (BONCAT) for the gut microbiota and combined it with fluorescently activated cell sorting and sequencing (FACS-Seq) to identify the translationally active members of the community. We then used this novel technique to compare and contrast to other methods of bulk community measurements of activity and viability: physiological staining of relative nucleic acid content and membrane damage. Relative nucleic acid content has previously been linked to metabolic activity, yet remains currently undefined for the human gut microbiota.ResultsTen healthy, unrelated individuals were sampled to determine the proportion and diversity of distinct physiological fractions of their gut microbiota. The translationally active bacteria represent about half of the gut microbiota, and are not distinct from the whole community. The high nucleic acid content (HNA) bacteria also represent about half of the gut microbiota, but are distinct from the whole community and correlate with the damaged subset. Perturbing the community with xenobiotics previously shown to alter bacterial activity but not diversity resulted in stronger changes in the distinct physiological fractions than in the whole community.ConclusionsBONCAT is a suitable method to probe the translationally active members of the human gut microbiota, and combined with FACS-Seq, allows for their identification. The high nucleic acid content bacteria are not necessarily the protein-producing bacteria in the community, and so further work is needed to understand the relationship between nucleic acid content and bacterial metabolism in the human gut. Taking into account physiologically distinct subsets of the gut microbiota may be more informative than relying on whole community profiling.

scholarly journals Effects of oxygen exposure on relative nucleic acid content and membrane integrity in the human gut microbiota

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e10602
Author(s):  
Mariia Taguer ◽  
Ophélie Quillier ◽  
Corinne F. Maurice
Keyword(s):  
Nucleic Acid ◽  
Gut Microbiota ◽  
Acid Content ◽  
Membrane Integrity ◽  
Nucleic Acid Content ◽  
Human Gut ◽  
Short Term ◽  
Bacterial Physiology ◽  
Oxygen Exposure ◽  
Anaerobic Chamber

While the diversity of the human gut microbiota is becoming increasingly well characterized, bacterial physiology is still a critical missing link in understanding how the gut microbiota may be implicated in disease. The current best practice for studying bacterial physiology involves the immediate storage of fecal samples in an anaerobic chamber. This reliance on immediate access to anaerobic chambers greatly limits the scope of sample populations that can be studied. Here, we assess the effects of short-term oxygen exposure on gut bacterial physiology and diversity. We use relative nucleic acid content and membrane integrity as markers of bacterial physiology, and 16S rRNA gene amplicon sequencing to measure bacterial diversity. Samples were stored for up to 6 h in either ambient conditions or in anoxic environments created with gas packs or in an anaerobic chamber. Our data indicate that AnaeroGen sachets preserve bacterial membrane integrity and nucleic acid content over the course of 6 h similar to storage in an anaerobic chamber. Short-term oxygen exposure increases bacterial membrane permeability, without exceeding inter-individual differences. As oxygen exposure remains an important experimental consideration for bacterial metabolism, our data suggest that AnaeroGen sachets are a valid alternative limiting loss of membrane integrity for short-term storage of samples from harder-to-access populations.

scholarly journals Using flow cytometry and light-induced fluorescence technique to characterizethe variability and characteristics of bioaerosols in springtime at Metro Atlanta, Georgia

2018 ◽  
Author(s):  
Arnaldo Negron ◽  
Natasha DeLeon-Rodriguez ◽  
Samantha M. Waters ◽  
Luke D. Ziemba ◽  
Bruce Anderson ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Nucleic Acid ◽  
Acid Content ◽  
Fungal Spores ◽  
Epifluorescence Microscopy ◽  
Nucleic Acid Content ◽  
Rain Event ◽  
Bacterial Cells ◽  
Sampling System ◽  
Spores And Pollen

Abstract. The abundance and speciation of primary biological aerosol particles (PBAP) is important for understanding their impacts on human health, cloud formation and ecosystems. Towards this, we have developed a protocol for quantifying PBAP collected from large volumes of air with a portable wet-walled cyclone bioaerosol sampler. A flow cytometry (FCM) protocol was then developed to quantify and characterize the PBAP populations from the sampler, which were confirmed against epifluorescence microscopy. The sampling system and FCM analysis were used to study PBAP in Atlanta, GA over a two-month period and showed clearly defined populations of DNA-containing particles: Low Nucleic Acid-content particles (bioLNA), High Nucleic Acid-content particles (HNA) being fungal spores and pollen. We find that daily-average springtime PBAP concentration (1 to 5 μm diameter) ranged between 1.4 × 104 and 1.1 × 105 m−3. The BioLNA population dominated PBAP during dry days (72 ± 18 %); HNA dominated the PBAP during humid days and following rain events, where HNA (e.g., wet-ejected fungal spores) comprised up to 92 % of the PBAP number. Concurrent measurements with a Wideband Integrated Bioaerosol Sensor (WIBS-4A) showed that FBAP and total FCM counts are similar; HNA (from FCM) significantly correlated with ABC type FBAP concentrations throughout the sampling period (and for the same particle size range, 1–5 μm diameter). However, the FCM bioLNA population, possibly containing bacterial cells, did not correlate to any FBAP type. The lack of correlation of any WIBS FBAP type with the bioLNA suggest bacterial cells may be more difficult to detect with autofluorescence than previously thought. Ιdentification of bacterial cells even in the FCM (bioLNA population) is challenging, given that the fluorescence level of stained cells at times may be comparable to that seen from abiotic particles. HNA and ABC displayed highest concentration on a humid and warm day after a rain event (4/14), suggesting that both populations correspond to wet-ejected fungal spores. Overall, information from both instruments combined reveals a highly dynamic airborne bioaerosol community over Atlanta, with a considerable presence of fungal spores during humid days, and a bioLNA population dominating bioaerosol community during dry days.

scholarly journals Titanium Dioxide Nanoparticles Elicit Lower Direct Inhibitory Effect on Human Gut Microbiota Than Silver Nanoparticles

2019 ◽  
Vol 172 (2) ◽  
pp. 411-416 ◽  
Author(s):  
Richard T Agans ◽  
Alex Gordon ◽  
Saber Hussain ◽  
Oleg Paliy
Keyword(s):  
Silver Nanoparticles ◽  
Titanium Dioxide ◽  
Gut Microbiota ◽  
Titanium Dioxide Nanoparticles ◽  
Close Association ◽  
Community Diversity ◽  
Bacterial Cells ◽  
Human Gut ◽  
Human Gut Microbiota

Abstract Due to continued technological development, people increasingly come in contact with engineered nanomaterials (ENMs) that are now used in foods and many industrial applications. Many ENMs have historically been shown to possess antimicrobial properties, which has sparked concern for how dietary nanomaterials impact gastrointestinal health via microbial dysbiosis. We employed an in vitro Human Gut Simulator system to examine interactions of dietary nano titanium dioxide (TiO2) with human gut microbiota. Electron microscopy indicated a close association of TiO2 particles with bacterial cells. Addition of TiO2 to microbial communities led to a modest reduction in community density but had no impact on community diversity and evenness. In contrast, administration of known antimicrobial silver nanoparticles (NPs) in a control experiment resulted in a drastic reduction of population density. In both cases, communities recovered once the addition of nanomaterials was ceased. Constrained ordination analysis of community profiles revealed that simulated colonic region was the primary determinant of microbiota composition. Accordingly, predicted community functional capacity and measured production of short-chain fatty acids were not changed significantly upon microbiota exposure to TiO2. We conclude that tested TiO2 NPs have limited direct effect on human gut microbiota.

scholarly journals Using flow cytometry and light-induced fluorescence to characterize the variability and characteristics of bioaerosols in springtime in Metro Atlanta, Georgia

2020 ◽  
Vol 20 (3) ◽  
pp. 1817-1838 ◽  
Author(s):  
Arnaldo Negron ◽  
Natasha DeLeon-Rodriguez ◽  
Samantha M. Waters ◽  
Luke D. Ziemba ◽  
Bruce Anderson ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Nucleic Acid ◽  
Acid Content ◽  
Aerosol Particles ◽  
Fungal Spores ◽  
Epifluorescence Microscopy ◽  
Nucleic Acid Content ◽  
Rain Event ◽  
Bacterial Cells ◽  
Biological Aerosol Particles

Abstract. The abundance and speciation of primary biological aerosol particles (PBAP) is important for understanding their impacts on human health, cloud formation, and ecosystems. Towards this, we have developed a protocol for quantifying PBAP collected from large volumes of air with a portable wet-walled cyclone bioaerosol sampler. A flow cytometry (FCM) protocol was then developed to quantify and characterize the PBAP populations from the sampler, which were confirmed against epifluorescence microscopy. The sampling system and FCM analysis were used to study PBAP in Atlanta, GA, over a 2-month period and showed clearly defined populations of nucleic-acid-containing particles: low nucleic acid-content particles above threshold (LNA-AT) and high nucleic acid-content particles (HNA) likely containing wet-ejected fungal spores and pollen. We find that the daily-average springtime PBAP concentration (1 to 5 µm diameter) ranged between 1.4×104 and 1.1×105 m−3. The LNA-AT population dominated PBAP during dry days (72±18 %); HNA dominated the PBAP during humid days and following rain events, where HNA comprised up to 92 % of the PBAP number. Concurrent measurements with a Wideband Integrated Bioaerosol Sensor (WIBS-4A) showed that fluorescent biological aerosol particles (FBAP) and total FCM counts are similar; HNA (from FCM) moderately correlated with ABC-type FBAP concentrations throughout the sampling period (and for the same particle size range, 1–5 µm diameter). However, the FCM LNA-AT population, possibly containing bacterial cells, did not correlate with any FBAP type. The lack of correlation of any WIBS FBAP type with the LNA-AT suggests that airborne bacterial cells may be more difficult to unambiguously detect with autofluorescence than currently thought. Identification of bacterial cells even in the FCM (LNA-AT population) is challenging, given that the fluorescence level of stained cells at times may be comparable to that seen from abiotic particles. HNA and ABC displayed the highest concentration on a humid and warm day after a rain event (14 April 2015), suggesting that both populations correspond to wet-ejected fungal spores. Overall, information from both instruments combined reveals a highly dynamic airborne bioaerosol community over Atlanta, with a considerable presence of fungal spores during humid days and an LNA-AT population dominating the bioaerosol community during dry days.

scholarly journals Does the High Nucleic Acid Content of Individual Bacterial Cells Allow Us To Discriminate between Active Cells and Inactive Cells in Aquatic Systems?

2001 ◽  
Vol 67 (4) ◽  
pp. 1775-1782 ◽  
Author(s):  
Philippe Lebaron ◽  
Pierre Servais ◽  
Helene Agogué ◽  
Claude Courties ◽  
Fabien Joux
Keyword(s):  
Flow Cytometry ◽  
Nucleic Acid ◽  
Acid Content ◽  
Fluorescent Dyes ◽  
Specific Activity ◽  
Aquatic Systems ◽  
Nucleic Acid Content ◽  
Leucine Incorporation ◽  
Bacterial Cells ◽  
Syto 13

ABSTRACT The nucleic acid contents of individual bacterial cells as determined with three different nucleic acid-specific fluorescent dyes (SYBR I, SYBR II, and SYTO 13) and flow cytometry were compared for different seawater samples. Similar fluorescence patterns were observed, and bacteria with high apparent nucleic acid contents (HNA) could be discriminated from bacteria with low nucleic acid contents (LNA). The best discrimination between HNA and LNA cells was found when cells were stained with SYBR II. Bacteria in different water samples collected from seven freshwater, brackish water, and seawater ecosystems were prelabeled with tritiated leucine and then stained with SYBR II. After labeling and staining, HNA, LNA, and total cells were sorted by flow cytometry, and the specific activity of each cellular category was determined from leucine incorporation rates. The HNA cells were responsible for most of the total bacterial production, and the specific activities of cells in the HNA population varied between samples by a factor of seven. We suggest that nucleic acid content alone can be a better indicator of the fraction of growing cells than total counts and that this approach should be combined with other fluorescent physiological probes to improve detection of the most active cells in aquatic systems.

scholarly journals Gut Microbiome and Social Determinants of Health (SDOH)

2020 ◽  
Vol 4 (2) ◽  
pp. 01-05
Author(s):  
Suneeta Kumari
Keyword(s):  
Gut Microbiota ◽  
Gut Microbiome ◽  
Bacterial Cells ◽  
Medical Field ◽  
Human Gut ◽  
Environmental Implications ◽  
Human Microbiota ◽  
The Social ◽  
Tremendous Amount

With technological advancements in the medical field, new discoveries have been unfolded about the human microbiota. A tremendous amount of work has been studied within the last two decades. Some of the human microbiota sites include nonsterile areas such as mouth, skin, gut, nose, and vagina. Additionally, there are bacterial cells in areas that were considered sterile such as lungs and placenta before delivery. Out of all the sites, the gut houses the most with an amount of 100 trillion bacteria (Guinane, 2013). Environmental implications have been known to impact these new areas of medicine. There has been a growing interest by the social epidemiologists on how health inequalities impact the role of human gut microbiota.

scholarly journals DETERMINING RELATIVE IMPORTANCE OF HUMAN GUT MICROBIOTA, AGE, GENDER AND LIFESTYLE PATTERN AS A PREDICTOR FOR BMI USING LOGMPIE DATA

2021 ◽  
Vol 21 (2) ◽  
Author(s):  
Komal Jani ◽  
Shelly Gupta
Keyword(s):  
Physical Activity ◽  
Body Mass Index ◽  
Random Forest ◽  
Gut Microbiota ◽  
Relative Abundance ◽  
Gut Microbiome ◽  
Relative Importance ◽  
Human Gut ◽  
Human Gut Microbiota ◽  
Lifestyle Pattern

We use the ‘Relative Abundance Table’ and ‘LogMPIE Study Metadata’ from the “Landscape of Gut Microbiome - Pan-India Exploration”, or LogMPIE dataset to find out the relative importance of human gut microbiota abundance (specifically genus), age, gender, and lifestyle pattern as a predictor for BMI (Body Mass Index). The LogMPIE data is taken from 1004 subjects and 993 unique microorganisms are reported along with BMI, age, and physical activity. We use Random Forest Regressor to find out the relative importance of the above-mentioned features (microorganism genus abundance, age, gender, and lifestyle pattern) in predicting the BMI of a subject. The objective here is not the prediction of BMI using the features but to find out the relative importance of these features as much as these affect the BMI.

scholarly journals The Human Gut Microbiota: A Key Mediator of Osteoporosis and Osteogenesis

2021 ◽  
Vol 22 (17) ◽  
pp. 9452
Author(s):  
Kevin D. Seely ◽  
Cody A. Kotelko ◽  
Hannah Douglas ◽  
Brandon Bealer ◽  
Amanda E. Brooks
Keyword(s):  
Gut Microbiota ◽  
Bone Healing ◽  
Gut Microbiome ◽  
Healing Process ◽  
Micro Rna ◽  
Bone Homeostasis ◽  
Human Gut ◽  
Surgical Interventions ◽  
Human Gut Microbiota ◽  
Therapeutic Consideration

An expanding body of research asserts that the gut microbiota has a role in bone metabolism and the pathogenesis of osteoporosis. This review considers the human gut microbiota composition and its role in osteoclastogenesis and the bone healing process, specifically in the case of osteoporosis. Although the natural physiologic processes of bone healing and the pathogenesis of osteoporosis and bone disease are now relatively well known, recent literature suggests that a healthy microbiome is tied to bone homeostasis. Nevertheless, the mechanism underlying this connection is still somewhat enigmatic. Based on the literature, a relationship between the microbiome, osteoblasts, osteoclasts, and receptor activator of nuclear factor-kappa-Β ligand (RANKL) is contemplated and explored in this review. Studies have proposed various mechanisms of gut microbiome interaction with osteoclastogenesis and bone health, including micro-RNA, insulin-like growth factor 1, and immune system mediation. However, alterations to the gut microbiome secondary to pharmaceutical and surgical interventions cannot be discounted and are discussed in the context of clinical therapeutic consideration. The literature on probiotics and their mechanisms of action is examined in the context of bone healing. The known and hypothesized interactions of common osteoporosis drugs and the human gut microbiome are examined. Since dysbiosis in the gut microbiota can function as a biomarker of bone metabolic activity, it may also be a pharmacological and nutraceutical (i.e., pre- and probiotics) therapeutic target to promote bone homeostasis.

scholarly journals Phylogenetic barriers to horizontal transfer of antimicrobial peptide resistance genes in the human gut microbiota

2018 ◽  
Author(s):  
Bálint Kintses ◽  
Orsolya Méhi ◽  
Eszter Ari ◽  
Mónika Számel ◽  
Ádám Györkei ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Gut Microbiota ◽  
Resistance Genes ◽  
Gut Microbiome ◽  
Horizontal Transfer ◽  
Bacterial Species ◽  
Antibiotic Resistance Genes ◽  
Genetic Exchange ◽  
Human Gut ◽  
Human Gut Microbiota

AbstractThe human gut microbiota has adapted to the presence of antimicrobial peptides (AMPs) that are ancient components of immune defence. Despite important medical relevance, it has remained unclear whether AMP resistance genes in the gut microbiome are available for genetic exchange between bacterial species. Here we show that AMP- and antibiotic-resistance genes differ in their mobilization patterns and functional compatibilities with new bacterial hosts. First, whereas AMP resistance genes are widespread in the gut microbiome, their rate of horizontal transfer is lower than that of antibiotic resistance genes. Second, gut microbiota culturing and functional metagenomics revealed that AMP resistance genes originating from phylogenetically distant bacteria only have a limited potential to confer resistance inEscherichia coli, an intrinsically susceptible species. Third, the phenotypic impact of acquired AMP resistance genes heavily depends on the genetic background of the recipient bacteria. Taken together, functional compatibility with the new bacterial host emerges as a key factor limiting the genetic exchange of AMP resistance genes. Finally, our results suggest that AMPs induce highly specific changes in the composition of the human microbiota with implications for disease risks.

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