The association between serum microbial DNA composition and symptoms of depression and anxiety in mood disorders

AbstractThere is increasing evidence supporting the association between gut microbiome composition and mood disorders; however, studies on the circulating microbiome are scarce. This study aimed to analyze the association of the serum microbial DNA composition with depressive and anxiety symptoms in patients with mood disorders. The sera of 69 patients with mood disorders, aged from 19 to 60, were analyzed. Bacterial DNA was isolated from extracellular membrane vesicles and, subsequently, amplified and quantified with specific primers for the V3–V4 hypervariable region of the 16S rDNA gene. Sequence reads were clustered into Operational Taxonomic Units and classified using the SILVA database. There were no significant associations between alpha diversity measures and the total Hamilton depression rating scale (HAM-D) or Beck anxiety inventory (BAI) scores. Only the weighted UniFrac distance was associated with the total HAM-D score (F = 1.57, p = 0.045). The Bacteroidaceae family and Bacteroides genus were negatively associated with the total HAM-D score (β = − 0.016, p < 0.001, q = 0.08 and β = − 0.016, p < 0.001, q = 0.15, respectively). The Desulfovibrionaceae family and Clostridiales Family XIII were positively associated with the total BAI score (β = 1.8 × 10−3, p < 0.001, q = 0.04 and β = 1.3 × 10−3, p < 0.001, q = 0.24, respectively). Further studies with larger sample sizes and longitudinal designs are warranted.

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Comprehensive Analysis of Risk Factors for Periodontitis Focusing on the Saliva Microbiome and Polymorphism

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph18126430 ◽

2021 ◽

Vol 18 (12) ◽

pp. 6430

Author(s):

Naoki Toyama ◽

Daisuke Ekuni ◽

Daisuke Matsui ◽

Teruhide Koyama ◽

Masahiro Nakatochi ◽

...

Keyword(s):

Genome Wide Association Study ◽

Alpha Diversity ◽

Study Data ◽

Shannon Index ◽

Rrna Gene ◽

Nucleotide Polymorphisms ◽

Unifrac Distance ◽

Microbiome Composition ◽

Significant Difference ◽

Unweighted Unifrac

Few studies have exhaustively assessed relationships among polymorphisms, the microbiome, and periodontitis. The objective of the present study was to assess associations simultaneously among polymorphisms, the microbiome, and periodontitis. We used propensity score matching with a 1:1 ratio to select subjects, and then 22 individuals (mean age ± standard deviation, 60.7 ± 9.9 years) were analyzed. After saliva collection, V3-4 regions of the 16S rRNA gene were sequenced to investigate microbiome composition, alpha diversity (Shannon index, Simpson index, Chao1, and abundance-based coverage estimator) and beta diversity using principal coordinate analysis (PCoA) based on weighted and unweighted UniFrac distances. A total of 51 single-nucleotide polymorphisms (SNPs) related to periodontitis were identified. The frequencies of SNPs were collected from Genome-Wide Association Study data. The PCoA of unweighted UniFrac distance showed a significant difference between periodontitis and control groups (p < 0.05). There were no significant differences in alpha diversity and PCoA of weighted UniFrac distance (p > 0.05). Two families (Lactobacillaceae and Desulfobulbaceae) and one species (Porphyromonas gingivalis) were observed only in the periodontitis group. No SNPs showed significant expression. These results suggest that periodontitis was related to the presence of P. gingivalis and the families Lactobacillaceae and Desulfobulbaceae but not SNPs.

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16s rRNA Lung Microbiota Study in Mechanically Ventilated Patient: A Pilot Study

10.21203/rs.3.rs-100088/v1 ◽

2020 ◽

Author(s):

Mélanie Fromentin ◽

Antoine Bridier-Nahmias ◽

Jérôme Legoff ◽

Severine Mercier-Delarue ◽

Noémie Ranger ◽

...

Keyword(s):

16S Rrna ◽

Hypervariable Region ◽

Human Microbiome ◽

Alpha Diversity ◽

Microbial Composition ◽

Optimal Method ◽

Unifrac Distance ◽

Bacterial Microbiota ◽

Paired End Sequencing ◽

16Srrna Gene

Abstract BackgroundCharacterization of the respiratory tract bacterial microbiota is in its infancy when compared to the gut microbiota knowledge. As key methodological steps can directly affect the accuracy of the results, it is crucial to determine a robust methodology in order to limit bias.Two different pairs of primers 515F-806R targeting the V4 hypervariable region of the 16SrRNA gene, “S-V4” for “Stringent V4” primer pair and “R-V4” for “Relaxed V4” primer pair, are respectively used in two ongoing international projects “the Human microbiome project” and “the Earth microbiome project”. We compared two methods of sample processing using these two different primer pairs for bacterial microbiota analyses of respiratory samples from critically-ill ventilated patients. For the later, Illumina 250 paired-end sequencing was done on a MiSeq platform after DNA extraction using mechanical lysis (bead-beating) and NucliSENS easyMAG. The concordance with conventional microbiology is the criterion of choice to determine the optimal method.ResultsTwenty samples from seven patients and four controls were sequenced. The two primer pair provided highly different results. Only 54% of the samples had a similar microbial composition with both pairs of primers. “S-V4” gave the best agreement with the conventional microbiology for ETA: 89% as compared to 44% for the “R-V4” primer pair. The main difference related to Enterobacteriaceae, which were concordant between conventional cultures and microbiota analyses using “S-V4”. Enterobacteriaceae detection was poor for “R-V4”. Among patients with VAP, a decrease in alpha diversity in ETA was observed. The mean of pairwise Unifrac distance was higher inside this group of patients at the time of VAP diagnosis as compared to control patients.ConclusionAccuracy of the bacterial lung microbiota composition was highly correlated to the pair of primers used for amplification of the 16s rRNA hypervariable sequence. Comparison of microbiota results obtained by sequencing and conventional microbiology allowed us to select the best option for further studies. This work validates our methodology based on 16SrRNA gene amplification with 515F-806R “S-V4” primer pair associated to Illumina® MiSeq 250 paired-end sequencing.

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Microbiome evaluation revealed salivary dysbiosis in addicts of betel nut preparations

10.1101/2020.04.13.20064063 ◽

2020 ◽

Author(s):

Faizan Saleem ◽

Ghulam Mujtaba ◽

Junaid Ahmed Kori ◽

Arshad Hassan ◽

M. Kamran Azim

Keyword(s):

Hypervariable Region ◽

Alpha Diversity ◽

Oral Microbiome ◽

P Value ◽

Rrna Gene ◽

Betel Nut ◽

Taxonomic Assignment ◽

Unifrac Distance ◽

Genus Level ◽

Slaked Lime

AbstractBetel nut addiction is recognized as the causative agent of oral microbiome dysbiosis and other systematic disorders. A number of betel nut preparations containing ingredients such as slaked lime, catechu extract and tobacco are being commonly used particularly in South Asia. The underlying variations in the oral microbiome due to usage of betel nut preparations are poorly understood. We evaluated salivary microbiome in response to chewing of betel nut preparation(s). In order to assess the microbiome dynamics, metagenomic analysis of 16S rRNA gene (V3-V4 hypervariable region) from salivary bacteria in chewers of betel nut preparation (n = 16) and non-chewers (n = 55) was carried out by Greengenes and SILVA ribosomal sequence databases. It was observed that Gutka chewers demonstrated lower alpha diversity and number of bacterial genera than the non-chewers. Taxonomic assignment on phylum level revealed Firmicutes (p-value = 0.042 at 95% confidence interval) to be significantly more abundant in Gutka chewers in comparison with non-chewers. Beta diversity analysis at genus level by weighted unifrac distance matrices unveiled both groups to be divergent from each other. On the genus level, Veillonella (p-value = 0.015), Streptococcus (p-value = 0.026), Leptotrichia (p-value = 0.022) and Serratia (p-value = 0.022) species appeared to be significantly more abundant in Gutka chewers in comparison to non-chewers. The present study suggests salivary dysbiosis in response to gutka chewing and concludes that gutka chewers possess higher abundance of acidogenic and aciduric bacteria. This study contributes additional information regarding oral microbiome variations with response to gutka consumption.

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Salivary Microbial Dysbiosis Is Associated With Peri-Implantitis: A Case-Control Study in a Brazilian Population

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.696432 ◽

2022 ◽

Vol 11 ◽

Author(s):

Debora Pallos ◽

Vanessa Sousa ◽

Magda Feres ◽

Belen Retamal-Valdes ◽

Tsute Chen ◽

...

Keyword(s):

Beta Diversity ◽

Bacterial Species ◽

Hypervariable Region ◽

Alpha Diversity ◽

Rrna Gene ◽

Microbial Composition ◽

Microbiome Composition ◽

Ion Torrent Pgm ◽

Microbial Dysbiosis ◽

Haemophilus Parainfluenzae

Background and ObjectivesThe aim of this study was to examine the salivary microbiome in healthy peri-implant sites and those with peri-implantitis.MethodsSaliva samples were collected from 21 participants with healthy peri-implant sites and 21 participants with peri-implantitis. The V4 hypervariable region of the 16S rRNA gene was sequenced using the Ion Torrent PGM System (Ion 318™ Chip v2 400). The NGS analysis and composition of the salivary microbiome were determined by taxonomy assignment. Downstream bioinformatic analyses were performed in QIIME (v 1.9.1).ResultsClinical differences according to peri-implant condition status were found. Alpha diversity metrics revealed that the bacterial communities of participants with healthy peri-implant sites tended to have a richer microbial composition than individuals with peri-implantitis. In terms of beta diversity, bleeding on probing (BoP) may influence the microbial diversity. However, no clear partitioning was noted between the salivary microbiome of volunteers with healthy peri-implant sites or volunteers with peri-implantitis. The highest relative abundance of Stenotrophomonas, Enterococcus and Leuconostoc genus, and Faecalibacterium prausnitzii, Haemophilus parainfluenzae, Prevotella copri, Bacteroides vulgatus, and Bacteroides stercoris bacterial species was found in participants with peri-implantitis when compared with those with healthy peri-implant sites.ConclusionDifferences in salivary microbiome composition were observed between patients with healthy peri-implant sites and those with peri-implantitis. BoP could affect the diversity (beta diversity) of the salivary microbiome.

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Gastrointestinal disturbance and effect of fecal microbiota transplantation in discharged COVID-19 patients

Journal of Medical Case Reports ◽

10.1186/s13256-020-02583-7 ◽

2021 ◽

Vol 15 (1) ◽

Author(s):

Fengqiong Liu ◽

Shanliang Ye ◽

Xin Zhu ◽

Xuesong He ◽

Shengzhou Wang ◽

...

Keyword(s):

B Cells ◽

Fecal Microbiota Transplantation ◽

Gastrointestinal Symptoms ◽

Fecal Microbiota ◽

Case Presentation ◽

Microbiome Composition ◽

Operational Taxonomic Units ◽

Gut Dysbiosis ◽

Rehabilitative Intervention ◽

Oral Capsule

Abstract Background To investigate the potential beneficial effect of fecal microbiota transplantation (FMT) on gastrointestinal symptoms, gut dysbiosis and immune status in discharged COVID-19 patients. Case presentation A total of 11 COVID-19 patients were recruited in April, 2020, about one month on average after they were discharged from the hospital. All subjects received FMT for 4 consecutive days by oral capsule administrations with 10 capsules for each day. In total, 5 out of 11 patients reported to be suffered from gastrointestinal symptoms, which were improved after FMT. After FMT, alterations of B cells were observed, which was characterized as decreased naive B cell (P = 0.012) and increased memory B cells (P = 0.001) and non-switched B cells (P = 0.012).The microbial community richness indicated by operational taxonomic units number, observed species and Chao1 estimator was marginally increased after FMT. Gut microbiome composition of discharged COVID-19 patients differed from that of the general population at both phylum and genera level, which was characterized with a lower proportion of Firmicutes (41.0%) and Actinobacteria (4.0%), higher proportion of Bacteroidetes (42.9%) and Proteobacteria (9.2%). FMT can partially restore the gut dysbiosis by increasing the relative abundance of Actinobacteria (15.0%) and reducing Proteobacteria (2.8%) at the phylum level. At the genera level, Bifidobacterium and Faecalibacterium had significantly increased after FMT. Conclusions After FMT, altered peripheral lymphocyte subset, restored gut microbiota and alleviated gastrointestinal disorders were observe, suggesting that FMT may serve as a potential therapeutic and rehabilitative intervention for the COVID-19.

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52 Alterations of fecal microbiome characteristics by dietary soy isoflavone ingestion in growing pigs infected with porcine reproductive and respiratory syndrome virus

Journal of Animal Science ◽

10.1093/jas/skaa054.054 ◽

2020 ◽

Vol 98 (Supplement_3) ◽

pp. 30-31

Author(s):

Brooke N Smith ◽

Stephen A Fleming ◽

Mei Wang ◽

Ryan N Dilger

Keyword(s):

Control Diet ◽

Growing Pigs ◽

Respiratory Syndrome Virus ◽

Post Inoculation ◽

Long Term Effects ◽

Unifrac Distance ◽

Fecal Microbiome ◽

Microbiome Composition ◽

Time Points ◽

Experimental Treatments

Abstract Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically-important disease and ingestion of soy isoflavones (ISF) may benefit PRRSV-infected pigs due to demonstrated anti-inflammatory and anti-viral properties. The objective of this study was to quantify long-term effects of ISF consumption on fecal microbiome characteristics under disease challenge. In total, 96 weaned barrows were group-housed in a BSL-2 containment facility and allotted to 1 of 3 experimental treatments that were maintained throughout the wean-to-finish study: non-infected pigs receiving an ISF-devoid control diet (NC, n=24), and infected pigs receiving either the control diet (PC, n=36) or that supplemented with total ISF in excess of 1,600 mg/kg (ISF, n=36) (Table 1). Following a 7-day adaptation, pigs were inoculated intranasally with either a sham-control (PBS) or live PRRSV (1×105 TCID50/mL, strain NADC20). Fecal samples were collected from 48 individual pigs at pre-infection (-2 days post-inoculation, DPI), peak-infection (10 DPI), and post-infection (144 DPI) time-points and extracted DNA was used for 16S bacterial rRNA sequencing. Differences in bacterial communities among diet groups were evaluated using UniFrac distance matrices (weighted and unweighted) in QIIME. All other data were analyzed by one-way ANOVA performed on transformed data using R. Across all time-points, only minimal differences were observed due to ISF alone. At 10 DPI, PRRSV infection reduced Prevotella 9 genera abundance from approximately 20% to less than 10%, but the specific function of this variety in pigs is unclear. The most notable finding was decreased relative abundance of Actinobacteria at 144 DPI between non-infected and infected treatments (P < 0.05), which is consistent with various dysbioses observed in other disease models. Our findings indicate that differences present were mainly due to PRRSV infection and not strongly influenced by ISF ingestion, which implies previously observed performance benefits conferred by dietary ISF are not likely due to changes in microbiome composition.

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The Effects of Genetic Relatedness on the Preterm Infant Gut Microbiota

Microorganisms ◽

10.3390/microorganisms9020278 ◽

2021 ◽

Vol 9 (2) ◽

pp. 278

Author(s):

Shen Jean Lim ◽

Miriam Aguilar-Lopez ◽

Christine Wetzel ◽

Samia V. O. Dutra ◽

Vanessa Bray ◽

...

Keyword(s):

Gut Microbiota ◽

Preterm Infant ◽

Neonatal Intensive Care ◽

Genetic Relatedness ◽

Bovine Milk ◽

Alpha Diversity ◽

Neonatal Intensive Care Units ◽

Rrna Gene ◽

Microbial Composition ◽

Operational Taxonomic Units

The preterm infant gut microbiota is influenced by environmental, endogenous, maternal, and genetic factors. Although siblings share similar gut microbial composition, it is not known how genetic relatedness affects alpha diversity and specific taxa abundances in preterm infants. We analyzed the 16S rRNA gene content of stool samples, ≤ and >3 weeks postnatal age, and clinical data from preterm multiplets and singletons at two Neonatal Intensive Care Units (NICUs), Tampa General Hospital (TGH; FL, USA) and Carle Hospital (IL, USA). Weeks on bovine milk-based fortifier (BMF) and weight gain velocity were significant predictors of alpha diversity. Alpha diversity between siblings were significantly correlated, particularly at ≤3 weeks postnatal age and in the TGH NICU, after controlling for clinical factors. Siblings shared higher gut microbial composition similarity compared to unrelated individuals. After residualizing against clinical covariates, 30 common operational taxonomic units were correlated between siblings across time points. These belonged to the bacterial classes Actinobacteria, Bacilli, Bacteroidia, Clostridia, Erysipelotrichia, and Negativicutes. Besides the influence of BMF and weight variables on the gut microbial diversity, our study identified gut microbial similarities between siblings that suggest genetic or shared maternal and environmental effects on the preterm infant gut microbiota.

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AB1232 ORAL DYSBIOSIS REFLECTS THE IMMUNOLOGICAL ALTERATION OF RA REGARDING TO ACPA AND HLA DRB1*SE: NAGASAKI ISLAND STUDY

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-eular.5147 ◽

2020 ◽

Vol 79 (Suppl 1) ◽

pp. 1907.2-1907

Author(s):

Y. Tsuji ◽

M. Tamai ◽

S. Morimoto ◽

D. Sasaki ◽

M. Nagayoshi ◽

...

Keyword(s):

Beta Diversity ◽

Healthy Subjects ◽

Microbial Community Composition ◽

Alpha Diversity ◽

Enzyme Linked Immunosorbent Assay ◽

Rrna Gene ◽

Oral Microbiota ◽

Unifrac Distance ◽

Alpha And Beta Diversity ◽

The Difference

Background:Anti-citrullinated protein antibody (ACPA) production is observed in several organs even prior to the onset of rheumatoid arthritis (RA), and oral mucosa is considered to be one of the important tissues. The presence of HLA-DRB1*SE closely associates with ACPA production. Saliva is considered to reflect the oral microbiota including periodontal disease. Alteration of oral microbiota of RA becomes to be normalized by DMARDs treatment, however, the interaction of HLA-DRB1*SE, ACPA and oral microbiota of RA patients remains to be elucidated.Objectives:The Nagasaki Island Study, which had started in 2014 collaborating with Goto City, is intended for research of the preclinical stage of RA, including ACPA/HLA genotype screening and ultrasound and magnetic resonance imaging examinations in high-risk subjects. Using the samples accumulated in this cohort, we have tried to investigate the difference of oral microbiota among RA patients and healthy subjects regarding to ACPA and HLA-DRB1*SE.Methods:Blood and salivary samples were obtained from 1422 subjects out of 4276 who have participated in the Nagasaki Island Study from 2016 to 2018. ACPA positivity was 1.7 % in total. Some of RA patients resided in Goto City participated in the Nagasaki Island Study. At this point, we selected 291 subjects, who were ACPA positive non-RA healthy subjects (n=22) and patients with RA (n=33, 11 subjects were ACPA positive and 22 ACPA negative respectively) as the case, age and gender matched ACPA negative non-RA healthy subjects (n=236) as the control. ACPA was measured by an enzyme-linked immunosorbent assay, and HLA genotyping was quantified by next-generation sequencing (Ref.1). The operational taxonomic unit (OUT) analysis using 16S rRNA gene sequencing were performed. The richness of microbial diversity within-subject (alpha diversity) was scaled via Shannon entropy. The dissimilarity between microbial community composition was calculated using Bray-Curtis distance as a scale, and differences between groups (beta diversity) were tested by permutational multivariate analysis of variance (PERMANOVA). In addition, UniFrac distance calculated in consideration of the distance on the phylogenetic tree were performed.Results:Median age 70 y.o., % Female 58.8 %. Among RA and non-RA subjects, not alpha diversity but beta diversity was statistically significance (p=0.022, small in RA). In RA subjects, both alpha and beta diversity is small (p<0.0001), especially significant in ACPA positive RA (Figure 1). Amongt RA subjects, presence of HLA-DRB1*SE did not show the difference but the tendency of being small of alpha diversity (p=0.29).Conclusion:Our study has suggested for the first time the association of oral microbiota alteration with the presence of ACPA and HLA-DRB1*SE. Oral dysbiosis may reflect the immunological status of patients with RA.References:[1]Kawaguchi S, et al. Methods Mol Biol 2018;1802: 22Disclosure of Interests:None declared

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Immune Gene Expression Covaries with Gut Microbiome Composition in Stickleback

mBio ◽

10.1128/mbio.00145-21 ◽

2021 ◽

Vol 12 (3) ◽

Author(s):

Lauren E. Fuess ◽

Stijn den Haan ◽

Fei Ling ◽

Jesse N. Weber ◽

Natalie C. Steinel ◽

...

Keyword(s):

Gene Expression ◽

Microbial Communities ◽

Gut Microbiome ◽

Alpha Diversity ◽

Host Immunity ◽

Host Gene ◽

Microbiota Composition ◽

Host Gene Expression ◽

Microbiome Composition ◽

Immune Genes

ABSTRACT Commensal microbial communities have immense effects on their vertebrate hosts, contributing to a number of physiological functions, as well as host fitness. In particular, host immunity is strongly linked to microbiota composition through poorly understood bi-directional links. Gene expression may be a potential mediator of these links between microbial communities and host function. However, few studies have investigated connections between microbiota composition and expression of host immune genes in complex systems. Here, we leverage a large study of laboratory-raised fish from the species Gasterosteus aculeatus (three-spined stickleback) to document correlations between gene expression and microbiome composition. First, we examined correlations between microbiome alpha diversity and gene expression. Our results demonstrate robust positive associations between microbial alpha diversity and expression of host immune genes. Next, we examined correlations between host gene expression and abundance of microbial taxa. We identified 15 microbial families that were highly correlated with host gene expression. These families were all tightly correlated with host expression of immune genes and processes, falling into one of three categories—those positively correlated, negatively correlated, and neutrally related to immune processes. Furthermore, we highlight several important immune processes that are commonly associated with the abundance of these taxa, including both macrophage and B cell functions. Further functional characterization of microbial taxa will help disentangle the mechanisms of the correlations described here. In sum, our study supports prevailing hypotheses of intimate links between host immunity and gut microbiome composition. IMPORTANCE Here, we document associations between host gene expression and gut microbiome composition in a nonmammalian vertebrate species. We highlight associations between expression of immune genes and both microbiome diversity and abundance of specific microbial taxa. These findings support other findings from model systems which have suggested that gut microbiome composition and host immunity are intimately linked. Furthermore, we demonstrate that these correlations are truly systemic; the gene expression detailed here was collected from an important fish immune organ (the head kidney) that is anatomically distant from the gut. This emphasizes the systemic impact of connections between gut microbiota and host immune function. Our work is a significant advancement in the understanding of immune-microbiome links in nonmodel, natural systems.

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Sputum Microbiome Composition in Patients With Squamous Cell Lung Carcinoma

10.21203/rs.3.rs-902707/v1 ◽

2021 ◽

Author(s):

E. D. Baranova ◽

V. G. Druzhinin ◽

L. V. Matskova ◽

P. S. Demenkov ◽

V. P . Volobaev ◽

...

Keyword(s):

Squamous Cell ◽

Inflammatory Responses ◽

Alpha Diversity ◽

Taxonomic Composition ◽

16S Rrna Genes ◽

Rrna Genes ◽

Cell Lung Carcinoma ◽

Microbiome Composition ◽

Β Diversity ◽

Genome Damage

Abstract Recent findings indicate that the microbiome can have a significant impact on the development of lung cancer by inducing inflammatory responses, causing dysbiosis and generating genome damage. The aim of this study was to search for bacterial markers of squamous cell carcinoma (LUSC). In the study, the taxonomic composition of the sputum microbiome of 40 men with untreated LUSC was compared with 40 healthy controls. Next Generation sequencing of bacterial 16S rRNA genes was used to determine the taxonomic composition of the respiratory microbiome. There was no differences in alpha diversity between the LUSC and control groups. Meanwhile, differences in the structure of bacterial communities (β diversity) among patients and controls differed significantly in sputum samples (pseudo-F = 1.65; p = 0.026). Only Streptococcus, Bacillus, Gemella and Haemophilus were found to be significantly increased in patients with LUSC compared to the control subjects, while 19 bacterial genera were significantly reduced, indicating a decrease in beta diversity in the microbiome of patients with LUSC. From our study, Streptococcus (Streptococcus agalactiae) emerges as the most likely LUSC biomarker, but more research is needed to confirm this assumption.

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