scholarly journals New Approaches for Isolation of Previously Uncultivated Oral Bacteria

2011 ◽  
Vol 78 (1) ◽  
pp. 194-203 ◽  
Author(s):  
M. V. Sizova ◽  
T. Hohmann ◽  
A. Hazen ◽  
B. J. Paster ◽  
S. R. Halem ◽  
...  
Keyword(s):  
Oral Cavity ◽  
Single Cell ◽  
Single Species ◽  
Oral Bacteria ◽  
Cell Cultivation ◽  
Human Oral Cavity ◽  
Successful Recovery ◽  
Number Of Microorganisms ◽  
Enrichment Techniques

ABSTRACTA significant number of microorganisms from the human oral cavity remain uncultivated. This is a major impediment to the study of human health since some of the uncultivated species may be involved in a variety of systemic diseases. We used a range of innovations previously developed to cultivate microorganisms from the human oral cavity, focusing on anaerobic species. These innovations include (i)in vivocultivation to specifically enrich for species actively growing in the oral cavity (the “minitrap” method), (ii) single-cell long-term cultivation to minimize the effect of fast-growing microorganisms, and (iii) modifications of conventional enrichment techniques, using media that did not contain sugar, including glucose. To enable cultivation of obligate anaerobes, we maintained strict anaerobic conditions in most of our cultivation experiments. We report that, on a per cell basis, the most successful recovery was achieved using minitrap enrichment (11%), followed by single-cell cultivation (3%) and conventional plating (1%). Taxonomically, the richest collection was obtained using the single-cell cultivation method, followed by minitrap and conventional enrichment, comprising representatives of 13, 9, and 4 genera, respectively. Interestingly, no single species was isolated by all three methods, indicating method complementarity. An important result is the isolation and maintenance in pure culture of 10 strains previously only known by their molecular signatures, as well as representatives of what are likely to be three new microbial genera. We conclude that the ensemble of new methods we introduced will likely help close the gap between cultivated and uncultivated species from the human oral cavity.

In vivo optical virtual biopsy of human oral cavity with harmonic generation microscopy

2010 ◽  
Author(s):  
M.-R. Tsai ◽  
S.-Y. Chen ◽  
D.-B. Shieh ◽  
P.-J. Lou ◽  
C.-K. Sun
Keyword(s):  
Oral Cavity ◽  
Harmonic Generation ◽  
Human Oral Cavity

scholarly journals Evidence of independent acquisition and adaption of ultra-small bacteria to human hosts across the highly diverse yet reduced genomes of the phylum Saccharibacteria

2018 ◽  
Author(s):  
Jeffrey S. McLean ◽  
Batbileg Bor ◽  
Thao T. To ◽  
Quanhui Liu ◽  
Kristopher A. Kerns ◽  
...  
Keyword(s):  
Oral Cavity ◽  
De Novo ◽  
Gc Content ◽  
Single Copy ◽  
Small Subunit ◽  
Rrna Gene ◽  
Ssu Rrna ◽  
Ssu Rrna Gene ◽  
Human Oral Cavity

ABSTRACTRecently, we discovered that a member of the Saccharibacteria/TM7 phylum (strain TM7x) isolated from the human oral cavity, has an ultra-small cell size (200-300nm), a highly reduced genome (705 Kbp) with limited de novo biosynthetic capabilities, and a very novel lifestyle as an obligate epibiont on the surface of another bacterium 1. There has been considerable interest in uncultivated phyla, particularly those that are now classified as the proposed candidate phyla radiation (CPR) reported to include 35 or more phyla and are estimated to make up nearly 15% of the domain Bacteria. Most members of the larger CPR group share genomic properties with Saccharibacteria including reduced genomes (<1Mbp) and lack of biosynthetic capabilities, yet to date, strain TM7x represents the only member of the CPR that has been cultivated and is one of only three CPR routinely detected in the human body. Through small subunit ribosomal RNA (SSU rRNA) gene surveys, members of the Saccharibacteria phylum are reported in many environments as well as within a diversity of host species and have been shown to increase dramatically in human oral and gut diseases. With a single copy of the 16S rRNA gene resolved on a few limited genomes, their absolute abundance is most often underestimated and their potential role in disease pathogenesis is therefore underappreciated. Despite being an obligate parasite dependent on other bacteria, six groups (G1-G6) are recognized using SSU rRNA gene phylogeny in the oral cavity alone. At present, only genomes from the G1 group, which includes related and remarkably syntenic environmental and human oral associated representatives1, have been uncovered to date. In this study we systematically captured the spectrum of known diversity in this phylum by reconstructing completely novel Class level genomes belonging to groups G3, G6 and G5 through cultivation enrichment and/or metagenomic binning from humans and mammalian rumen. Additional genomes for representatives of G1 were also obtained from modern oral plaque and ancient dental calculus. Comparative analysis revealed remarkable divergence in the host-associated members across this phylum. Within the human oral cavity alone, variation in as much as 70% of the genes from nearest oral clade (AAI 50%) as well as wide GC content variation is evident in these newly captured divergent members (G3, G5 and G6) with no environmental relatives. Comparative analyses suggest independent episodes of transmission of these TM7 groups into humans and convergent evolution of several key functions during adaptation within hosts. In addition, we provide evidence from in vivo collected samples that each of these major groups are ultra-small in size and are found attached to larger cells.

scholarly journals Single-Cell Genomics and the Oral Microbiome

2020 ◽  
Vol 99 (6) ◽  
pp. 613-620 ◽  
Author(s):  
M. Balachandran ◽  
K.L. Cross ◽  
M. Podar
Keyword(s):  
Microbial Community ◽  
Single Cell ◽  
Single Cells ◽  
Oral Bacteria ◽  
Oral Microbiome ◽  
Oral Microbiota ◽  
Oral Diseases ◽  
Interspecies Interactions ◽  
Single Cell Genomics

The human oral cavity is one of the first environments where microbes have been discovered and studied since the dawn of microbiology. Nevertheless, approximately 200 types of bacteria from the oral microbiota have remained uncultured in the laboratory. Some are associated with a healthy oral microbial community, while others are linked to oral diseases, from dental caries to gum disease. Single-cell genomics has enabled inferences on the physiology, virulence, and evolution of such uncultured microorganisms and has further enabled isolation and cultivation of several novel oral bacteria, including the discovery of novel interspecies interactions. This review summarizes some of the more recent advances in this field, which is rapidly moving toward physiologic characterization of single cells and ultimately cultivation of the yet uncultured. A combination of traditional microbiological approaches with genomic-based physiologic predictions and isolation strategies may lead to the oral microbiome being the first complex microbial community to have all its members cultivable in the laboratory. Studying the biology of the individual microbes when in association with other members of the community, in controlled laboratory conditions and in vivo, should lead to a better understanding of oral dysbiosis and its prevention and reversion.

scholarly journals Multiple Single-Cell Genomes Provide Insight into Functions of Uncultured Deltaproteobacteria in the Human Oral Cavity

PLoS ONE ◽  
2013 ◽  
Vol 8 (3) ◽  
pp. e59361 ◽  
Author(s):  
Alisha G. Campbell ◽  
James H. Campbell ◽  
Patrick Schwientek ◽  
Tanja Woyke ◽  
Alexander Sczyrba ◽  
...  
Keyword(s):  
Oral Cavity ◽  
Single Cell ◽  
Human Oral Cavity ◽  
Insight Into

scholarly journals Integrated Single-Cell Atlases Reveal an Oral SARS-CoV-2 Infection and Transmission Axis

2020 ◽  
Author(s):  
Ni Huang ◽  
Paola Perez ◽  
Takafumi Kato ◽  
Yu Mikami ◽  
Kenichi Okuda ◽  
...  
Keyword(s):  
Oral Cavity ◽  
Single Cell ◽  
Salivary Glands ◽  
Viral Entry ◽  
Immune Cell ◽  
Data Sets ◽  
Sequencing Data ◽  
Viral Shedding ◽  
Autopsy Tissues

ABSTRACTDespite signs of infection, the involvement of the oral cavity in COVID-19 is poorly understood. To address this, single-cell RNA sequencing data-sets were integrated from human minor salivary glands and gingiva to identify 11 epithelial, 7 mesenchymal, and 15 immune cell clusters. Analysis of SARS-CoV-2 viral entry factor expression showed enrichment in epithelia including the ducts and acini of the salivary glands and the suprabasal cells of the mucosae. COVID-19 autopsy tissues confirmed in vivo SARS-CoV-2 infection in the salivary glands and mucosa. Saliva from SARS-CoV-2-infected individuals harbored epithelial cells exhibiting ACE2 expression and SARS-CoV-2 RNA. Matched nasopharyngeal and saliva samples found distinct viral shedding dynamics and viral burden in saliva correlated with COVID-19 symptoms including taste loss. Upon recovery, this cohort exhibited salivary antibodies against SARS-CoV-2 proteins. Collectively, the oral cavity represents a robust site for COVID-19 infection and implicates saliva in viral transmission.

Preliminary study on in-vivo dielectric properties of tissues in the human oral cavity at microwave frequencies

2021 ◽  
Author(s):  
Donevollon Sladden ◽  
Lourdes Farrugia ◽  
Roberta Scicluna ◽  
Julian Bonello ◽  
Charles Sammut
Keyword(s):  
Dielectric Properties ◽  
Oral Cavity ◽  
Microwave Frequencies ◽  
Human Oral Cavity ◽  
Preliminary Study

scholarly journals In Vivo Drug Dissolution in Human Oral Cavity from Orally Disintegrating Tablet and Comparability with in Vitro Testing

2018 ◽  
Vol 66 (10) ◽  
pp. 999-1005 ◽  
Author(s):  
Tsuyoshi Katayama ◽  
Shinya Uchida ◽  
Chiaki Kamiya ◽  
Shimako Tanaka ◽  
Yasuharu Kashiwagura ◽  
...  
Keyword(s):  
Oral Cavity ◽  
Drug Dissolution ◽  
Orally Disintegrating Tablet ◽  
Human Oral Cavity

Nutritional Influences on Biofilm Development

1997 ◽  
Vol 11 (1) ◽  
pp. 81-99 ◽  
Author(s):  
G.H.W. Bowden ◽  
Y.H. Li
Keyword(s):  
Oral Cavity ◽  
Bacterial Species ◽  
Oral Bacteria ◽  
Carbon Limitation ◽  
Cell Surfaces ◽  
Nutritional Influences ◽  
Oral Biofilms ◽  
Biofilm Development

The amounts and types of nutrients in the environment influence the development and final bacterial and chemical composition of biofilms. In oligotrophic environments, organisms respond to nutrient stress by alterations in their cell morphology and cell surfaces, which enhance adherence. Little is known of the responses to stress by bacteria in the animal oral cavity. The environment in the oral cavity is less extreme, and saliva provides a constant source of nutrients. Catabolic cooperation among oral bacteria allows carbon and nitrogen from salivary glycoproteins to be utilized. Modification of growth environments of oral bacteria can influence their cell surfaces and adhesion. Studies in experimental animals have shown that feeding either glucose or sucrose diets or fasting has little effect on the initial stages of development of oral biofilms. However, diet can influence the proportions of different bacterial species later in biofilm development. Studies of competition among populations in communities of oral bacteria in vitro and in vivo have shown the significance of carbon limitation and excess and changes in environmental pH. Relatively few studies have been made of the role of a nitrogen metabolism in bacterial competition in biofilms. In keeping with biofilms in nature, oral biofilms provide a sequestered habitat, where organisms are protected from removal by saliva and where interactions among cells generate a biofilm environment, distinct from that of saliva. Oral biofilms are an essential component in the etiologies of caries and periodontal disease, and understanding the biology of oral biofilms has aided and will continue to aid in the prevention and treatment of these diseases.

scholarly journals Quercetin Preserves Oral Cavity Health by Mitigating Inflammation and Microbial Dysbiosis

2021 ◽  
Vol 12 ◽  
Author(s):  
Erin C. Mooney ◽  
Sara E. Holden ◽  
Xia-Juan Xia ◽  
Yajie Li ◽  
Min Jiang ◽  
...  
Keyword(s):  
Oral Cavity ◽  
Alveolar Bone ◽  
Treated Group ◽  
Oral Bacteria ◽  
Inflammatory Cell Infiltrate ◽  
Microbial Composition ◽  
Tlr Agonists ◽  
Microbial Dysbiosis

Failure to attenuate inflammation coupled with consequent microbiota changes drives the development of bone-destructive periodontitis. Quercetin, a plant-derived polyphenolic flavonoid, has been linked with health benefits in both humans and animals. Using a systematic approach, we investigated the effect of orally delivered Quercetin on host inflammatory response, oral microbial composition and periodontal disease phenotype. In vivo, quercetin supplementation diminished gingival cytokine expression, inflammatory cell infiltrate and alveolar bone loss. Microbiome analyses revealed a healthier oral microbial composition in Quercetin-treated versus vehicle-treated group characterized by reduction in the number of pathogenic species including Enterococcus, Neisseria and Pseudomonas and increase in the number of non-pathogenic Streptococcus sp. and bacterial diversity. In vitro, Quercetin diminished inflammatory cytokine production through modulating NF-κB:A20 axis in human macrophages following challenge with oral bacteria and TLR agonists. Collectively, our findings reveal that Quercetin supplement instigates a balanced periodontal tissue homeostasis through limiting inflammation and fostering an oral cavity microenvironment conducive of symbiotic microbiota associated with health. This proof of concept study provides key evidence for translational studies to improve overall health.

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