O-143 Characterization of vaginal and endometrial microbiome in patients with chronic endometritis (CE)

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
P LOZANO ◽  
A Bernabeu ◽  
B Lledó ◽  
R Morales ◽  
F I Aranda ◽  
...  
Keyword(s):  
16S Rrna ◽  
Embryo Transfer ◽  
Relative Abundance ◽  
Beta Diversity ◽  
Alpha Diversity ◽  
Shannon Index ◽  
Small Sample ◽  
Diagnostic Methods ◽  
Vaginal Microbiome ◽  
Chronic Endometritis

Abstract Study question Could vaginal and endometrial microbiome by sequencing 16S rRNA be comparable to classic diagnostic methods or immunohistochemistry CD138 for diagnosis of chronic endometritis? Summary answer A characteristic endometrial and vaginal microbiome is present in patients with chronic endometritis. An abnormal vaginal microbiome correlates with the presence of chronic endometritis. What is known already Chronic endometritis is a disease characterized by persistent inflammation of the endometrial lining. Currently, histopathological evaluation by immunohistochemistry CD138 marker is most common diagnostic method for CE. Microbiome analysis based on subunit 16S rRNA sequencing is a fast tool that can enable the identification of pathogenic microorganisms associated with CE. The main bacteria at vaginal and endometrial level belong to genus Lactobacillus, producers of lactic acid that allows maintaining acidic pH of vagina and acts as barrier against pathogens. Investigations on the effect of an abnormal endometrial and vaginal microbiome could improve assisted reproductive technologies. Study design, size, duration This is a observational pilot study (60 patients and 120 samples). The study population consists of patients attending to our fertility clinic for frozen euploid embryo transfer (FET) from May 2017 to May 2019. Preimplantation Genetic Testing of aneuploidy (PGT-A) was performed at blastocyst stage using Veriseq (Illumina). The inclusion criteria to be meet by patients were: age between 18 and 50 years, own or donated oocytes and use of ICSI. Participants/materials, setting, methods Cohort study with sixty patients undergoing assisted reproductive treatment (TRA) with their own or donated gametes and PGT-A Vaginal and endometrial samples were taken in the cycle prior to embryo transfer. The vaginal and endometrial microbiome was analyzed by mass sequencing of the V3V4 region of 16S rRNA. Bioinformatics analysis was performed using QIIME2 and MicrobiomeAnalyst packages. Alpha, beta diversity and taxonomic characterization were compared with positive and negative CD138 groups for chronic endometritis (CE). Main results and the role of chance Different bacterial communities were detected when vaginal and endometrial samples were analyzed in patients with and without endometritis diagnosed with CD138 immunohistochemistry. In patients with endometritis, a higher alpha diversity index tendency was found in vaginal samples (p = 0.15 for the Shannon index) and significant differences in endometrial samples (p = 0.01 for the Shannon index). In the beta diversity analysis, no significant differences were observed between the groups established as per the diagnosis of endometritis. Vaginal and endometrial samples from women with endometritis showed a microbiome pattern not dominated by Lactobacillus spp. Relative abundance analysis identified the genera Ralstonia and Gardnerella in endometrial sample, and the genera Streptoccoccus and Ureaplasma in vaginal sample of patients diagnosed with CD138 for endometritis. Comparing endometrial and vaginal samples CD138 positive diagnosed for endometritis, alpha diversity (p = 0.06 for the Shannon index and p = 0.08 for the Simpson index) and beta diversity (p < 0.001) showed significant differences. Relative abundance identified the genera Lactobacillus (p = 3.76E-4), Ralstonia (p = 8.19E-4), Delftia (p = 0.004) and Anaerobacillus (p = 0.004) in these sample groups. Limitations, reasons for caution The main limitation of this study is the small sample size. Larger studies including a higher number of samples are needed to confirm the different microbiome pattern observed at the vaginal and endometrial levels in correlation with chronic endometritis. The microbiome pattern has not been analyzed after treatment of CE. Wider implications of the findings Our findings suggest the existence of a characteristic vaginal and endometrial microbiota in patients with chronic endometritis. Different genera and species were identified in patients with and without endometritis depending on whether the sample was endometrial or vaginal. An abnormal vaginal microbiome appears to be strongly correlated with chronic endometritis. Trial registration number Not Applicable

scholarly journals The Impact Of The Vaginal And Endometrial Microbiome Pattern On Assisted Reproduction Outcomes

2020 ◽  
Author(s):  
Concepcion Carratala ◽  
María Carmen Díaz Martínez ◽  
Andrea Bernabeu García ◽  
Belén Lledó Bosch ◽  
José Antonio Quesada Rico ◽  
...  
Keyword(s):  
Assisted Reproduction ◽  
Relative Abundance ◽  
Beta Diversity ◽  
Alpha Diversity ◽  
Microbial Colonization ◽  
Control Group ◽  
Implantation Failure ◽  
Vaginal Microbiome ◽  
Failure Group ◽  
The Impact

Abstract Background:The vaginal microbiome plays an important role in maintaining health, and there is evidence that microbial colonization of the upper genital tract can also influence successful embryo transfer. The aim of this study is to determine whether the vaginal and endometrial microbiome in people undergoing assisted reproduction techniques could affect the pregnancy rate.Results:Regarding the microbiome dynamics during the cycle, we observed a decrease in alpha diversity from the follicular to luteal phase in the control group, in contrast to a stable pattern in the repetitive implantation failure group. As for endometrial and vaginal microbiome, alpha diversity was higher in the endometrium (Shannon p = 0.0139, Simpson p = 0.046); differences were also observed in beta diversity (p = 0.001). Compared to the endometrium, the vagina showed a greater relative abundance of Lactobacillus spp. (83.17% vs 84.82%, p < 0.0001), Streptococcus spp. (1.59% vs 7.74%, p = 0.014) and Ureaplasma spp. (0% vs. 0.89%, p = 0.006), and a lower abundance of Delftia spp. (0.95% vs 0%, p = 0.0003), Anaerobacillus spp. (1.59% vs 0%, p = 0.0004), and Ralstonia spp. (3.17% vs 0%, p = 0.0006). We also observed differences in both alpha diversity (Shannon p = 0.0206, Simpson p = 0.0206) and beta diversity between groups, along with differences for Ralstonia spp. (0.09% study group and 0.73% control, p = 0.0012). Finally, the relative abundance of Lactobacillus spp. differed between patients that did not versus did achieve pregnancy (91% vs 99%, p = 0.0445 visit 1, 94.63% vs. 97.69%, p = 0.0268 visit 2, 97.73% vs 99.74%, p = 0.0492 visit 3). The relative abundance of L.reuteri was also different between groups (0.39% vs 0.17%, p = 0.0397 visit 1, 0.15% vs 0.30%, p = 0.0491 visit 3).Conclusions:The vaginal and endometrial microbiome pattern correlates with the pregnancy rate, and it is different in patients who do versus do not have repetitive implantation failures. No significant differences in the composition of the microbiome were observed through the different visits. The lack of dynamism in the microbiome pattern of repetitive implantation failure patients might reflect an impaired adaptation to endometrial changes. A greater relative abundance of Lactobacillus spp. and L. reuteri is correlated with higher chances of pregnancy.*MC Díaz Martínez and A Bernabeu García share equal responsibility for authorship.

scholarly journals Ranking the Biases: The Choice of OTUs vs. ASVs in 16S rRNA Amplicon Data Analysis Has Stronger Effects On Diversity Measures Than Rarefaction and Similarity Threshold

2021 ◽  
Author(s):  
Marlène Chiarello ◽  
Mark McCauley ◽  
Sébastien Villéger ◽  
Colin R Jackson
Keyword(s):  
16S Rrna ◽  
Beta Diversity ◽  
Alpha Diversity ◽  
Diversity Indices ◽  
Taxonomic Composition ◽  
Shannon Index ◽  
Minimal Impact ◽  
Alpha And Beta Diversity ◽  
Methodological Choices ◽  
Similarity Threshold

Abstract BackgroundAdvances in the analysis of amplicon sequence datasets have introduced a methodological shift in how research teams investigate microbial biodiversity, away from the classification and downstream analyses of traditional operational taxonomic units (OTUs), and towards the usage of amplicon sequence variants (ASVs). While ASVs have several inherent properties that make them desirable compared to OTUs, questions remain as to the influence that these pipelines have on the ecological patterns being assessed, especially when compared to other methodological choices made when processing data (e.g. rarefaction) and computing diversity indices. ResultsWe compared the respective influences of using ASVs vs. OTU-based pipelines, rarefaction of the community table, and OTU similarity threshold (97% vs. 99%) on the ecological signals detected in freshwater invertebrate and environmental (sediment, seston) 16S rRNA data sets, determining the effects on alpha diversity, beta diversity and taxonomic composition. While the choice of OTU vs. ASV pipeline significantly influenced unweighted alpha and beta diversities and changed the ecological signal detected, weighted indices such as the Shannon index, Bray-Curtis dissimilarity, and weighted Unifrac scores were not impacted by the pipeline followed. By comparison, OTU threshold and rarefaction had a minimal impact effect on all measurements, although rarefaction improved overall signals, especially in OTU-based datasets. The identification of major classes and genera identified revealed significant discrepancies across methodologies. ConclusionWe provide a list of recommendations for the analysis of 16S rRNA amplicon data. We notably recommend the use of ASVs when analyzing alpha-diversity patterns, especially in species-rich or environmental samples. Abundance weighted alpha- and beta-diversity indices should also be preferred compared to ones based on the presence-absence of biological units.

scholarly journals The Diversity, Composition, and Metabolic Pathways of Archaea in Pigs

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 2139
Author(s):  
Feilong Deng ◽  
Yushan Li ◽  
Yunjuan Peng ◽  
Xiaoyuan Wei ◽  
Xiaofan Wang ◽  
...  
Keyword(s):  
Relative Abundance ◽  
Antibiotic Resistance Genes ◽  
Alpha Diversity ◽  
Distance Matrix ◽  
Archaeal Community ◽  
Gene Families ◽  
Shannon Index ◽  
Metabolic Potential ◽  
Substantial Progress ◽  
Archaeal Communities

Archaea are an essential class of gut microorganisms in humans and animals. Despite the substantial progress in gut microbiome research in the last decade, most studies have focused on bacteria, and little is known about archaea in mammals. In this study, we investigated the composition, diversity, and functional potential of gut archaeal communities in pigs by re-analyzing a published metagenomic dataset including a total of 276 fecal samples from three countries: China (n = 76), Denmark (n = 100), and France (n = 100). For alpha diversity (Shannon Index) of the archaeal communities, Chinese pigs were less diverse than Danish and French pigs (p < 0.001). Consistently, Chinese pigs also possessed different archaeal community structures from the other two groups based on the Bray–Curtis distance matrix. Methanobrevibacter was the most dominant archaeal genus in Chinese pigs (44.94%) and French pigs (15.41%), while Candidatus methanomethylophilus was the most predominant in Danish pigs (15.71%). At the species level, the relative abundance of Candidatus methanomethylophilus alvus, Natrialbaceae archaeon XQ INN 246, and Methanobrevibacter gottschalkii were greatest in Danish, French, and Chinese pigs with a relative abundance of 14.32, 11.67, and 16.28%, respectively. In terms of metabolic potential, the top three pathways in the archaeal communities included the MetaCyc pathway related to the biosynthesis of L-valine, L-isoleucine, and isobutanol. Interestingly, the pathway related to hydrogen consumption (METHANOGENESIS-PWY) was only observed in archaeal reads, while the pathways participating in hydrogen production (FERMENTATION-PWY and PWY4LZ-257) were only detected in bacterial reads. Archaeal communities also possessed CAZyme gene families, with the top five being AA3, GH43, GT2, AA6, and CE9. In terms of antibiotic resistance genes (ARGs), the class of multidrug resistance was the most abundant ARG, accounting for 87.41% of archaeal ARG hits. Our study reveals the diverse composition and metabolic functions of archaea in pigs, suggesting that archaea might play important roles in swine nutrition and metabolism.

Fecal Microbiota Diversity in Survivors of Adolescent/Young Adult Hodgkin Lymphoma

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 1533-1533
Author(s):  
Wendy Cozen ◽  
Guoqin Yu ◽  
Mitchell Gail ◽  
Bharat N. Nathwani ◽  
Amie E. Hwang ◽  
...  
Keyword(s):  
Young Adult ◽  
Gut Microbiota ◽  
Hodgkin Lymphoma ◽  
Relative Abundance ◽  
Alpha Diversity ◽  
Fecal Microbiota ◽  
Shannon Index ◽  
Tree Index ◽  
Unique Otus ◽  
Whole Tree

Abstract Abstract 1533 Survivors of adolescent/young adult Hodgkin lymphoma (AYAHL) report fewer exposures to infections during childhood compared to controls. They also have persistent genomic and functional aberrations in their lymphocytes that are partially attributable to chemotherapy or radiotherapy. Recent studies have shown that the gut microbiome can affect both the innate and adaptive immune response, and can suppress or exacerbate an inflammatory response. Given the central role of the gut microbiota in immune function, we investigated whether AYAHL survivors, who were members of 13 mono- and dizygotic twin pairs discordant for this disease, have differences in the diversity or phylogenetic configurations of their fecal microbiota compared to their unaffected co-twins. Twin pairs discordant for AYAHL are an ideal study population because they are at least partially matched on genetic and early life factors, both of which influence the composition of the gut microbiome. Pyrosequencing of bacterial 16S rRNA amplicons generated from single fecal samples obtained from each individual yielded 253,182 filtered and de-noised reads translated into species-level operational taxonomic units (OTUs). Standardized across individuals by random sampling, reads were assigned to 2513 OTUs to compare microbiome diversity and relative abundance of taxa. The number of OTU's was compared between twins using a paired student's t-test and a one-way analysis of variance was performed to determine whether such measures differed across twin pairs by comparing the measures between twins to those of randomly paired individuals. AYAHL survivors had less diverse fecal microbial communities compared to their unaffected co-twin controls by all measures of alpha diversity (Table 1). Measures that weighted the relative abundance of the bacteria were not statistically significantly different (Shannon Index, p= 0.270; Chao index, p= 0.066, PD Whole Tree Index, p= 0.051). However, when the unweighted number of unique OTUs was considered, the difference was significant (338 in cases vs. 369 in unaffected co-twin controls, p= 0.015). When the analysis was restricted to OTUs that were present at an abundance of > 0.1% in at least 2 of the 23 samples analyzed, the differences were attenuated, with only the PD Whole Tree index difference in diversity remaining marginally significant (p= 0.045). Only one bacterial taxon was associated with AYAHL, probably due to chance. Phylogenetic measurements indicated that the bacterial component of the microbiota of co-twins were more similar with respect to one another than unrelated individuals, although no differences by zygosity were observed. These results provide evidence that AYAHL survivors have reduced diversity of the gut microbiota, perhaps as a consequence the disease, its treatment, or a particularly hygienic environment. Table 1. Comparisons of alpha diversity measurements between Hodgkin lymphoma cases and co-twin controls. Measurements of Alpha Diversity Mean (Cases) Mean (Unaffected Co-twins) Mean Difference (Unaffected co-twin-case difference) P-value1 Initial analysis No. unique OTUs 338 369 31 0.015 Shannon index 5.6 5.8 0.2 0.27 Chao1 533 574 41 0.066 PD_whole tree 21.2 22.8 1.6 0.051 Conservative analysis No. unique OTUs 183 196 13 0.10 Shannon index 5.2 5.4 0.2 0.40 Chao1 230 237 7 0.47 PD_whole tree 13.7 14.6 0.9 0.045 1 P-value by paired t-tests. Disclosures: No relevant conflicts of interest to declare.

scholarly journals 2576. The Microbiome of Recurrent Bacterial Vaginosis Compared with Asymptomatic Controls

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S895-S895
Author(s):  
Elizabeth O Shay ◽  
Oluwatosin Goje ◽  
Roshan Padmanabhan ◽  
Charis Eng
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Bacterial Vaginosis ◽  
Alpha Diversity ◽  
Dna Probe ◽  
Gardnerella Vaginalis ◽  
Rrna Gene ◽  
Vaginal Microbiome ◽  
Β Diversity ◽  
Probe Test

Abstract Background Bacterial vaginosis (BV) affects nearly 1 in 3 women in the United States and is poorly understood. The study of the vaginal microbiome, using 16S rRNA-gene amplicon sequencing, has increased our knowledge of BV. We aimed to characterize the vaginal microbiome of women with recurrent BV firstly in comparison to controls, and secondly in comparison to a sub-population of our asymptomatic controls, positive for Gardnerella vaginalis via a vaginal pathogens DNA direct probe test (DNA probe). Methods Women aged 18–40 years, with recurrent BV, and asymptomatic controls were prospectively enrolled. Vaginal samples were collected from each participant. DNA was extracted, amplified using primers targeting the V3-V4 variable region of the 16S rRNA-gene, and then sequenced and processed through a hybrid Qiime MICCA bioinformatics pipeline. We also tested for G. vaginalis using the DNA probe. Results Seventeen recurrent BV patients and 46 controls were enrolled. Β diversity (P = 0.045), but not alpha diversity (P = 0.076) differed between groups. The genera Gardnerella and Prevotella were relatively more abundant, while Lactobacillus was relatively less abundant in recurrent BV vs. control groups. Of the patients for whom results of the DNA probe for Gardnerella vaginalis were available, 11 (69%) recurrent BV patients and 14 (35%) controls were positive. Control patients, negative by the DNA probe test, showed decreased alpha diversity (P = 0.0001) and significantly different β diversity (P = 0.001) compared with recurrent BV patients. Neither alpha (P = 0.31) nor β (P = 0.096) diversity differed between recurrent BV patients and controls that were G. vaginalis positive. Conclusion The microbiome of recurrent BV patients is distinct from that of asymptomatic controls; recurrent BV patients exhibit different β diversity, less Lactobacillus and more Gardnerella and Prevotella. Asymptomatic Gardnerella vaginalis-colonized controls demonstrate similar microbiome profiles to those of recurrent BV patients. These findings suggest that individual factors may influence whether or not a patient with a BV microbiomic profile experiences symptoms. Further investigation into these mechanisms could yield insights into the treatment of recurrent BV. Disclosures All authors: No reported disclosures.

scholarly journals Complementing 16S rRNA Gene Amplicon Sequencing with Total Bacterial Load To Infer Absolute Species Concentrations in the Vaginal Microbiome

mSystems ◽  
2020 ◽  
Vol 5 (2) ◽  
Author(s):  
Florencia A. Tettamanti Boshier ◽  
Sujatha Srinivasan ◽  
Anthony Lopez ◽  
Noah G. Hoffman ◽  
Sean Proll ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Relative Abundance ◽  
Bacterial Species ◽  
Bacterial Load ◽  
Amplicon Sequencing ◽  
Individual Species ◽  
Rrna Gene ◽  
Vaginal Microbiome ◽  
Bacterial Concentrations

ABSTRACT Whereas 16S rRNA gene amplicon sequencing quantifies relative abundances of bacterial taxa, variation in total bacterial load between samples restricts its ability to reflect absolute concentrations of individual bacterial species. Quantitative PCR (qPCR) can quantify individual species, but it is not practical to develop a suite of qPCR assays for every bacterium present in a diverse sample. We sought to determine the accuracy of an inferred measure of bacterial concentration using total bacterial load and relative abundance. We analyzed 1,320 samples from 20 women with a history of frequent bacterial vaginosis who self-collected vaginal swabs daily over 60 days. We inferred bacterial concentrations by taking the product of species relative abundance (assessed by 16S rRNA gene amplicon sequencing) and bacterial load (measured by broad-range 16S rRNA gene qPCR). Log10-converted inferred concentrations correlated with targeted qPCR (r = 0. 935, P < 2.2e–16) for seven key bacterial species. The mean inferred concentration error varied across bacteria, with rarer bacteria associated with larger errors. A total of 92% of the >0.5-log10 errors occurred when the relative abundance was <10%. Many errors occurred during early bacterial expansion from or late contraction to low abundance. When the relative abundance of a species is >10%, inferred concentrations are reliable proxies for targeted qPCR in the vaginal microbiome. However, targeted qPCR is required to capture bacteria at low relative abundance and is preferable for characterizing growth and decay kinetics of single species. IMPORTANCE Microbiome studies primarily use 16S rRNA gene amplicon sequencing to assess the relative abundance of bacterial taxa in a community. However, these measurements do not accurately reflect absolute taxon concentrations. We sought to determine whether the product of species’ relative abundance and total bacterial load measured by broad-range qPCR is an accurate proxy for individual species’ concentrations, as measured by taxon-specific qPCR assays. Overall, the inferred bacterial concentrations were a reasonable proxy of species-specific qPCR values, particularly when bacteria are present at a higher relative abundance. This approach offers an opportunity to assess the concentrations of bacterial species and how they change in a community over time without developing individual qPCR assays for each taxon.

scholarly journals Measuring Alpha and Beta Diversity by Field and Remote-Sensing Data: A Challenge for Coastal Dunes Biodiversity Monitoring

2021 ◽  
Vol 13 (10) ◽  
pp. 1928
Author(s):  
Flavio Marzialetti ◽  
Silvia Cascone ◽  
Ludovico Frate ◽  
Mirko Di Febbraro ◽  
Alicia Teresa Rosario Acosta ◽  
...  
Keyword(s):  
Remote Sensing ◽  
Beta Diversity ◽  
Central Italy ◽  
Alpha Diversity ◽  
Coastal Dunes ◽  
Shannon Index ◽  
Distance Measures ◽  
Spectral Variability ◽  
Floristic Similarity ◽  
The Relationship

Combining field collected and remotely sensed (RS) data represents one of the most promising approaches for an extensive and up-to-date ecosystem assessment. We investigated the potential of the so called spectral variability hypothesis (SVH) in linking field-collected and remote-sensed data in Mediterranean coastal dunes and explored if spectral diversity provides reliable information to monitor floristic diversity, as well as the consistency of such information in altered ecosystems due to plant invasions. We analyzed alpha diversity and beta diversity, integrating floristic field and Remote-Sensing PlanetScope data in the Tyrrhenian coast (Central Italy). We explored the relationship among alpha field diversity (species richness, Shannon index, inverse Simpson index) and spectral variability (distance from the spectral centroid index) through linear regressions. For beta diversity, we implemented a distance decay model (DDM) relating field pairwise (Jaccard similarities index, Bray–Curtis similarities index) and spectral pairwise (Euclidean distance) measures. We observed a positive relationship between alpha diversity and spectral heterogeneity with richness reporting the higher R score. As for DDM, we found a significant relationship between Bray–Curtis floristic similarity and Euclidean spectral distance. We provided a first assessment of the relationship between floristic and spectral RS diversity in Mediterranean coastal dune habitats (i.e., natural or invaded). SVH provided evidence about the potential of RS for estimating diversity in complex and dynamic landscapes.

scholarly journals Inconsistent Patterns of Microbial Diversity and Composition Between Highly Similar Sequencing Protocols: A Case Study With Reef-Building Corals

2021 ◽  
Vol 12 ◽  
Author(s):  
Hannah E. Epstein ◽  
Alejandra Hernandez-Agreda ◽  
Samuel Starko ◽  
Julia K. Baum ◽  
Rebecca Vega Thurber
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Beta Diversity ◽  
Sequence Data ◽  
Alpha Diversity ◽  
Gene Profiling ◽  
Rrna Gene ◽  
Sequencing Platform ◽  
Alpha And Beta Diversity ◽  
Diversity Metrics

16S rRNA gene profiling (amplicon sequencing) is a popular technique for understanding host-associated and environmental microbial communities. Most protocols for sequencing amplicon libraries follow a standardized pipeline that can differ slightly depending on laboratory facility and user. Given that the same variable region of the 16S gene is targeted, it is generally accepted that sequencing output from differing protocols are comparable and this assumption underlies our ability to identify universal patterns in microbial dynamics through meta-analyses. However, discrepant results from a combined 16S rRNA gene dataset prepared by two labs whose protocols differed only in DNA polymerase and sequencing platform led us to scrutinize the outputs and challenge the idea of confidently combining them for standard microbiome analysis. Using technical replicates of reef-building coral samples from two species, Montipora aequituberculata and Porites lobata, we evaluated the consistency of alpha and beta diversity metrics between data resulting from these highly similar protocols. While we found minimal variation in alpha diversity between platform, significant differences were revealed with most beta diversity metrics, dependent on host species. These inconsistencies persisted following removal of low abundance taxa and when comparing across higher taxonomic levels, suggesting that bacterial community differences associated with sequencing protocol are likely to be context dependent and difficult to correct without extensive validation work. The results of this study encourage caution in the statistical comparison and interpretation of studies that combine rRNA gene sequence data from distinct protocols and point to a need for further work identifying mechanistic causes of these observed differences.

scholarly journals Resident microbes of lactation rooms and daycares

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e8168
Author(s):  
Diana H. Taft ◽  
Samir Akre ◽  
Nicolas Madrid ◽  
Andre Knoesen ◽  
David A. Mills ◽  
...  
Keyword(s):  
Beta Diversity ◽  
Alpha Diversity ◽  
Illumina Miseq ◽  
Shannon Index ◽  
Rrna Gene ◽  
Sample Collection ◽  
Unifrac Distance ◽  
Modern Development ◽  
Temperature And Humidity ◽  
The Impact

Dedicated lactation rooms are a modern development as mothers return to work while still providing breastmilk to their absent infants. This study describes the built environment microbiome of lactation rooms and daycares, and explores the influence of temperature and humidity on the microbiome of lactation rooms. Sterile swabs were used to collect samples from five different sites in lactation rooms at University of California, Davis and from five different sites in daycares located in Davis, California. DNA from the swabs was extracted and the V4 region of the 16S rRNA gene was sequenced using Illumina MiSeq. Temperature and relative humidity data were collected on a subset of the lactation rooms. Sampled lactation rooms could be either dedicated lactation rooms or could also serve other functions (e.g., combined lactation room and restroom lounge). The majority of sequence reads were identified as belonging to family Moraxellaceae, with 73% of all reads included in analysis identified as an unknown species of Acinetobacter. Alpha diversity was analyzed using the Shannon index, while beta diversity was analyzed using unweighted and weighted UniFrac distance. The Jaccard distance was used to measure amount of change at sampling locations between time points for analysis of the impact of temperature and humidity on the microbiome. There were significant differences in the beta diversity of the microbiome of lactation rooms by room type. There were also significant differences in the beta diversity of the microbiome by sample collection location. There were no significant differences in either alpha or beta diversity associated with room temperature or humidity. Additional studies are needed to understand if the differences in lactation room type may result in differences in the breastmilk microbiome of milk collected in those rooms, and to what extent any such differences may influence the infant microbiome.

scholarly journals Lactobacillus pentosus KCA1 Decreases Vaginal and Gut Microbiota Associated with Bacterial Vaginosis (BV), Down-regulates IL-1 beta in Women of Child-Bearing Age and Modulates Bacterial Genes Related to Metabolic Functions

2020 ◽  
Author(s):  
Kingsley C Anukam ◽  
Chinwe E Ejike ◽  
Nneka R Agbakoba ◽  
Charlotte B Oguejiofor
Keyword(s):  
16S Rrna ◽  
Gut Microbiota ◽  
Bacterial Vaginosis ◽  
Relative Abundance ◽  
Beta Diversity ◽  
Oral Feeding ◽  
Lactobacillus Pentosus ◽  
Metabolic Functions ◽  
Before And After ◽  
Bacterial Genes

ABSTRACTIntroductionBacterial vaginosis which affects 14-50% of reproductive-aged women in Nigeria is misdiagnosed and under-reported. Treatment option is antibiotics, which leads to recurrent infections. The objectives of this study are three folds, first to determine effects of oral feeding of Lactobacillus pentosus KCA1 on the vaginal and gut microbiota of women diagnosed with BV; to measure the level of two proinflammatory cytokines IL-1 beta, before and after KCA1 consumption and to determine the relative abundance of bacterial metabolic genes.MethodsSeven women diagnosed with BV by Nugent score (7-10) were recruited to provide vaginal and gut sample before and after 14 days oral intake of 3 grams of Lactobacillus pentosus KCA1. The DNA from the swabs were processed for 16S rRNA metagenomics using Illumina MiSeq platform. The paired-end sequence FASTQ reads were imported into Illumina Basespace pipeline for quality check (QC). In addition, EzBioCloud pipeline was use for alpha and beta diversity estimation using PKSSU4.0 version and open reference UCLUST_MC2 for OTUs picking at 97% cut-off. Blood samples were analyzed using ELISA technique. PICRUSt was used to predict the metabolic functions from the 16S rRNA gene dataset.ResultsOn average, there was no significant difference at p=0.05 in the alpha indices typified by Shannon index. The beta diversity showed different clustering positions with PCoA. However, at individual taxonomic categories, there was a significant decrease in the relative abundance of some genera associated with bacterial vaginosis after KCA1 feeding with a corresponding increase of Lactobacillus genera. Bacterial genes related to defence systems were up-regulated in the vagina. There was a 2-fold down-regulation of IL-1 beta after consumption of KCA1.ConclusionOur findings suggest that Lactobacillus pentosus KCA1 taken orally, lowers pro-inflammatory cytokine, IL-1 beta and decreases the relative abundance of BV-associated bacteria.

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