scholarly journals 16S Metagenomics investigation of Bacterial diversity and prediction of its functionalities in Moroccan phosphate mine ecosystem.

2021 ◽  
Author(s):  
Salah Eddine Azaroual ◽  
Yassine Kasmi ◽  
Abderrahim Aasfar ◽  
Hicham El Arroussi ◽  
Youssef Zeroual ◽  
...  
Keyword(s):  
16S Rrna ◽  
Bacterial Diversity ◽  
Bacterial Communities ◽  
Native Plants ◽  
Extreme Environments ◽  
Wild Plants ◽  
Taxonomic Resolution ◽  
Rrna Gene ◽  
Hypervariable Regions ◽  
Phosphate Mine

Abstract Background Native plants in extreme environments may harbor some unique microbial communities with particular functions to sustain their growth and tolerance to harsh conditions. The aim of this study was to investigate the bacterial communities profiles in the Moroccan phosphate mine using 16S metagenomics sequencing. Taxonomic resolution of six hypervariable regions of the 16S rRNA was assessed in the collected samples and mock standard community. The bacterial functions enriched in the samples were also predicted to assess the contribution of the proposed low cost approach in the description of bacterial diversity and functionality in soil and plant. Methods The Ion 16S™ metagenomics kit was used to compare the discriminatory potential of the six targeted hypervariable regions of the 16S rRNA gene in the description of the bacterial communities. Alpha and beta diversity analysis were also performed accordingly in the studied samples. The later were collected from the bulk phosphate mine and the soils attached to roots of three different wild plants, that natively grow in this environment. Samples from the rhizosphere of wheat plants in cropping system were also studied. Actinobacteria related sequences were compared between the samples to investigate their possible origin between the bulk mine and the wild plants rhizosphere. The prediction of 82 bacterial functions related to nitrogen and phosphate metabolism along with stress was performed using Tax4fun bioinformatics tool. Results The rhizosphere of the three wild plants in the Moroccan phosphate mine is characterized by interesting bacterial diversity including Proteobacteria (62,24%, 71,15% and 65,61%), Actinobacteria (22,53%, 15,24%, 22,30%), Bacteroidetes (7,57% ; 4,23% ; 7,63%), and Firmicutes (5,82%; 1,17% ; 2,83%). Broad taxonomic diversity of minor phyla was also found in the native plants samples and included Acidobacteria, Armatimonadetes, Chloroflexi, Cyanobacteria, Gemmatimonadetes and Verrucomicrobia. The bulk phosphate mine samples were dominated by Actinobacteria with average relative abundance of 97,73% and Proteobacteria was Less abundant phylum in these samples. V3, V4 and V67 regions performed better in the taxonomic resolution at different levels e.g., phylum, family and genus. However, V8 and V9 regions gave the lesser taxonomic resolution and V9 region identified only Proteobacteria phylum in the mock standard sample. Alpha and beta diversity analysis indicated low abundance of observed OTUs and diversity in the bulk samples compared with the rhizosphere samples of the native plants in the phosphate mine. Moreover, significant (p = 0,021) differences in bacterial diversity were observed among at least three kind of samples : The bulk phosphate mine, the rhizosphere of the native wild plants of the phosphate mine and the rhizosphere of the wheat crop. The results indicated that both plant genotype and mainly soil conditions may be involved in the shaping of bacterial diversity. Such indication was also confirmed by the prediction of functional profiles that showed enrichment of many functions related to biological nitrogen fixation in the rhizosphere of native plants and the stress related functions in the bulk phosphate mine in comparison with the wheat rhizosphere samples Conclusion The proposed amplicon sequencing approach combined to bioinformatic prediction of bacterial functions could be considered as low-cost method to study the bacterial communities in soils and plants. The application of such approach to samples from the phosphate mines and wild plants revealed the need to consider at least V3, V4 and V6-7 hypervariable regions of the 16S rRNA gene in the taxonomic resolution of bacterial diversity in soil samples. This approach revealed also interesting bacterial diversity governed by the plant genotypes and the local conditions with interesting predicted functionalities involved in plant growth and development and stress tolerance in extreme environments. Further studies could target some interesting identified phyla and predicted bacterial functionalities, such studies could also deeply investigate the origin of the bacterial diversity in these native plants to develop new generation of bacterial inoculants for sustainable agriculture.

scholarly journals The impact of persistent bacterial bronchitis on the pulmonary microbiome of children

2017 ◽  
Author(s):  
Leah Cuthbertson ◽  
Vanessa Craven ◽  
Lynne Bingle ◽  
William O.C.M. Cookson ◽  
Mark L. Everard ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Bacterial Diversity ◽  
Community Composition ◽  
Bacterial Communities ◽  
Microbial Community Analysis ◽  
Rrna Gene ◽  
Healthy Children ◽  
Significant Difference ◽  
The Impact

AbstractPersistent bacterial bronchitis is a leading cause of chronic wet cough in young children. This study aimed to characterise the respiratory bacterial microbiota of healthy children and to assess the impact of the changes associated with the development of persistent bacterial bronchitis.Blind, protected brushings were obtained from 20 healthy controls and 24 children with persistent bacterial bronchitis, with an additional directed sample obtained from persistent bacterial bronchitis patients. DNA was extracted, quantified using a 16S rRNA gene quantitative PCR assay prior to microbial community analysis by 16S rRNA gene sequencing.No significant difference in bacterial diversity or community composition (R2 = 0.01, P = 0.36) was observed between paired blind and non-blind brushes, showing that blind brushings are a valid means of accessing the airway microbiota. This has important implications for collecting lower respiratory samples from healthy children. A significant decrease in bacterial diversity (P < 0.001) and change in community composition (R2 = 0.08, P = 0.004) was observed between controls and patients. Bacterial communities within patients with PBB were dominated by Proteobacteria, and indicator species analysis showed that Haemophilus and Neisseria were significantly associated with the patient group. In 15 (52.9%) cases the dominant organism by sequencing was not identified by standard routine clinical culture.The bacteria present in the lungs of patients with persistent bacterial bronchitis were less diverse in terms of richness and evenness. The results validate the clinical diagnosis, and suggest that more attention to bacterial communities in children with chronic cough may lead to more rapid recognition of this condition with earlier treatment and reduction in disease burden.

scholarly journals Bacterial Communities in Aquatic Sediment from Bukit Duabelas Raintforest and Oil Palm Plantation at Sumatra Indonesia

2018 ◽  
Vol 25 (2) ◽  
pp. 85
Author(s):  
Marini Wijayanti ◽  
Aris Tri Wahyudi ◽  
Munti Yuhana ◽  
Martin Engelhaupt ◽  
Anja Meryandini
Keyword(s):  
16S Rrna ◽  
Bacterial Diversity ◽  
Oil Palm ◽  
Bacterial Communities ◽  
Total Carbon ◽  
Available Phosphorus ◽  
Rrna Gene ◽  
Oil Palm Plantation ◽  
Community Shift ◽  
Aquatic Sediment

The diversity of bacterial communities in aquatic sediment of rainforest and oil palm plantation at Sumatra was studied using pyrosequencing of 16S rRNA gene and common biodiversity indices.  Phylogenetic approach was used for revealing the community shift of bacterial phyla and genera in both areas.  Ecological approach used soil pH, total Carbon (TC), total Nitrogen (TN), available Phosphorus (AP) measurement and bacterial diversity with Shanon and Simpson indices, and bacterial richness with Chao1-ACE indices and OTUs.  Bacterial diversity and richness in aquatic sediment of forest area was higher than that ones in oil palm plantation area, although their pH, TC, and TN in both areas were not different significantly. The majority of sequences related to Proteobacteria (34.85%), Acidobacteria (32.67%), Nitrospirae (6.86%), Chloroflexi (4.31%), and Actinobacteria (4.02%) were from forest; whereas those related to Acidobacteria (46.10%), Proteobacteria (25.86%), Nitrospirae (9.20%), Chloroflexi (4.99%), and Actinobacteria (2.34%) invented from oil palm plantation. The genera of Alphaproteobacteria and Betaproteobacteria dominated in genera phylotype of bacterial 16S rRNA phylogenetic revealed both aquatic sediment of forest and oil palm plantation. The most genera in the phylogenetic tree from aquatic sediment of both areas was Burkholderia.  The bacterial community shift in aquatic sediment of forest transformation indicated higher bacterial diversity index, richness index, some of phyla and genera in aquatic sediment from forest than from oil palm plantation.

Molecular characterization of the bacterial communities present in sheep's milk and cheese produced in South Brazilian Region via 16S rRNA gene metabarcoding sequencing

LWT ◽  
2021 ◽  
Vol 147 ◽  
pp. 111579
Author(s):  
Creciana M. Endres ◽  
Ícaro Maia S. Castro ◽  
Laura D. Trevisol ◽  
Juliana M. Severo ◽  
Michele B. Mann ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Characterization ◽  
Bacterial Communities ◽  
Rrna Gene

scholarly journals Sponge-Associated Bacteria Are Strictly Maintained in Two Closely Related but Geographically Distant Sponge Hosts

2011 ◽  
Vol 77 (20) ◽  
pp. 7207-7216 ◽  
Author(s):  
Naomi F. Montalvo ◽  
Russell T. Hill
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Bacterial Communities ◽  
Sponge Species ◽  
Rrna Gene ◽  
Gene Sequences ◽  
Associated Bacteria ◽  
Content Type ◽  
Key Species ◽  
Sponge Associated Bacteria

ABSTRACTThe giant barrel spongesXestospongiamutaandXestospongiatestudinariaare ubiquitous in tropical reefs of the Atlantic and Pacific Oceans, respectively. They are key species in their respective environments and are hosts to diverse assemblages of bacteria. These two closely related sponges from different oceans provide a unique opportunity to examine the evolution of sponge-associated bacterial communities. Mitochondrial cytochrome oxidase subunit I gene sequences fromX.mutaandX.testudinariashowed little divergence between the two species. A detailed analysis of the bacterial communities associated with these sponges, comprising over 900 full-length 16S rRNA gene sequences, revealed remarkable similarity in the bacterial communities of the two species. Both sponge-associated communities include sequences found only in the twoXestospongiaspecies, as well as sequences found also in other sponge species and are dominated by three bacterial groups,Chloroflexi,Acidobacteria, andActinobacteria. While these groups consistently dominate the bacterial communities revealed by 16S rRNA gene-based analysis of sponge-associated bacteria, the depth of sequencing undertaken in this study revealed clades of bacteria specifically associated with each of the twoXestospongiaspecies, and also with the genusXestospongia, that have not been found associated with other sponge species or other ecosystems. This study, comparing the bacterial communities associated with closely related but geographically distant sponge hosts, gives new insight into the intimate relationships between marine sponges and some of their bacterial symbionts.

scholarly journals Diversity of Culturable Actinomycetes from Deepsea Floor of Makassar Strait, Indonesia

2018 ◽  
Vol 3 (2) ◽  
pp. 73
Author(s):  
Ariani Hatmanti ◽  
Puspita Lisdiyanti ◽  
Jaka Widada ◽  
Subagus Wahyuono
Keyword(s):  
16S Rrna ◽  
Gene Sequence ◽  
Sodium Dodecyl ◽  
Extreme Environments ◽  
Rrna Gene ◽  
Rrna Gene Sequence ◽  
Track Record ◽  
Centrifugation Method ◽  
Isolation Media ◽  
Active Substances

With regard to collaboration research called Widya Nusantara Exploration (EWIN) in May-June 2013 and November 2014, a study on isolation of actinomycetes from sediments of Makassar Strait have been conducted. Actinomycetes is one of microbe which has an excellent track record in producing antimikrob and other active substances. But due to terrestrial actinomycetes has been widely explored, then recently researchers began focusing on wide variety of extreme environments, such as marine environment, to screening aktinomisetes in producing new secondary metabolites. A total of 36 strains of actinomycetes were isolated from 10 samples obtained from deepsea floor in Makassar Strait, Indonesia, Direct Dillution Method were best used to isolate the actinomycetes compare to Sodium Dodecyl Sulfida – Yeast Extract Method (SDS-YE Method) and Rehidration Centrifugation Method (RC Method). NBRC-802 media and Actinomycetes Isolation Agar(AIA)(Himedia)media were used as the isolation media. All the isolates were identified by morphological characteristic and by analysis of 16S rRNA gene sequence. Actinomycetes isolated from deepsea floor of Makassar Strait have been dominated by Micromonospora (58%), Verrucosispora (14%)Streptomyces (8%) and Luteipulveratus (5%), however genus Nocardiopsis, Micrococcus, Gordonia, Kytococcus, and Arthrobacter were not dominant (3%). Station 25 in 1.547 m depth was the most abundant of actinomycetes, 18 strains and dominated by the genus Micromonospora which is isolated using Direct Dillution Method and both NBRC 802 or AIA media.

scholarly journals Diversity and Structure of the Endophytic Bacterial Communities Associated With Three Terrestrial Orchid Species as Revealed by 16S rRNA Gene Metabarcoding

2020 ◽  
Vol 11 ◽  
Author(s):  
Pasquale Alibrandi ◽  
Sylvia Schnell ◽  
Silvia Perotto ◽  
Massimiliano Cardinale
Keyword(s):  
Bacterial Community ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Host Plant ◽  
Bacterial Communities ◽  
Reproductive Organs ◽  
Distribution Model ◽  
Rrna Gene ◽  
Orchid Species ◽  
Core Microbiota

The endophytic microbiota can establish mutualistic or commensalistic interactions within the host plant tissues. We investigated the bacterial endophytic microbiota in three species of Mediterranean orchids (Neottia ovata, Serapias vomeracea, and Spiranthes spiralis) by metabarcoding of the 16S rRNA gene. We examined whether the different orchid species and organs, both underground and aboveground, influenced the endophytic bacterial communities. A total of 1,930 operational taxonomic units (OTUs) were obtained, mainly Proteobacteria and Actinobacteria, whose distribution model indicated that the plant organ was the main determinant of the bacterial community structure. The co-occurrence network was not modular, suggesting a relative homogeneity of the microbiota between both plant species and organs. Moreover, the decrease in species richness and diversity in the aerial vegetative organs may indicate a filtering effect by the host plant. We identified four hub OTUs, three of them already reported as plant-associated taxa (Pseudoxanthomonas, Rhizobium, and Mitsuaria), whereas Thermus was an unusual member of the plant microbiota. Core microbiota analysis revealed a selective and systemic ascent of bacterial communities from the vegetative to the reproductive organs. The core microbiota was also maintained in the S. spiralis seeds, suggesting a potential vertical transfer of the microbiota. Surprisingly, some S. spiralis seed samples displayed a very rich endophytic microbiota, with a large number of OTUs shared with the roots, a situation that may lead to a putative restoring process of the root-associated microbiota in the progeny. Our results indicate that the bacterial community has adapted to colonize the orchid organs selectively and systemically, suggesting an active involvement in the orchid holobiont.

scholarly journals Microbial Diversity Profiling of Gut Microbiota of Macropus giganteus Using Three Hypervariable Regions of the Bacterial 16S rRNA

2021 ◽  
Vol 9 (8) ◽  
pp. 1721
Author(s):  
Christian O’Dea ◽  
Roger Huerlimann ◽  
Nicole Masters ◽  
Anna Kuballa ◽  
Cameron Veal ◽  
...  
Keyword(s):  
16S Rrna ◽  
Gut Microbiota ◽  
Microbial Diversity ◽  
Surface Waters ◽  
Bacterial Species ◽  
Rrna Gene ◽  
Faecal Contamination ◽  
Hypervariable Regions ◽  
Geographical Regions ◽  
V3 Region

Animal faecal contamination of surface waters poses a human health risk, as they may contain pathogenic bacteria or viruses. Of the numerous animal species residing along surface waterways in Australia, macropod species are a top contributor to wild animals’ faecal pollution load. We characterised the gut microbiota of 30 native Australian Eastern Grey Kangaroos from six geographical regions (five kangaroos from each region) within South East Queensland in order to establish their bacterial diversity and identify potential novel species-specific bacteria for the rapid detection of faecal contamination of surface waters by these animals. Using three hypervariable regions (HVRs) of the 16S rRNA gene (i.e., V1–V3, V3–V4, and V5–V6), for their effectiveness in delineating the gut microbial diversity, faecal samples from each region were pooled and microbial genomic DNA was extracted, sequenced, and analysed. Results indicated that V1-V3 yielded a higher taxa richness due to its larger target region (~480 bp); however, higher levels of unassigned taxa were observed using the V1-V3 region. In contrast, the V3–V4 HVR (~569 bp) attained a higher likelihood of a taxonomic hit identity to the bacterial species level, with a 5-fold decrease in unassigned taxa. There were distinct dissimilarities in beta diversity between the regions, with the V1-V3 region displaying the highest number of unique taxa (n = 42), followed by V3–V4 (n = 11) and V5–V6 (n = 8). Variations in the gut microbial diversity profiles of kangaroos from different regions were also observed, which indicates that environmental factors may impact the microbial development and, thus, the composition of the gut microbiome of these animals.

scholarly journals A pilot study of the effect of deployment on the gut microbiome and traveler’s diarrhea susceptibility

2020 ◽  
Author(s):  
Blake W. Stamps ◽  
Wanda J. Lyon ◽  
Adam P. Irvin ◽  
Nancy Kelley-Loughnane ◽  
Michael S. Goodson
Keyword(s):  
Pilot Study ◽  
16S Rrna ◽  
Gut Microbiome ◽  
16S Rrna Gene Sequencing ◽  
Initial Study ◽  
Mitigation Strategies ◽  
Taxonomic Resolution ◽  
Rrna Gene ◽  
Traveler’S Diarrhea ◽  
Traveler's Diarrhea

AbstractTraveler’s diarrhea (TD) is a recurrent and significant issue for many travelers including the military. While many known enteric pathogens exist that are causative agents of diarrhea, our gut microbiome may also play a role in travelers’ diarrhea susceptibility. To this end we conducted a pilot study of the microbiome of warfighters prior to- and after deployment overseas to identify marker taxa relevant to traveler’s diarrhea. This initial study utilized full-length 16S rRNA gene sequencing to provide additional taxonomic resolution towards identifying predictive taxa.16S rRNA analyses of pre- and post-deployment fecal samples identified multiple marker taxa as significantly differentially abundant in subjects that reported diarrhea, including Weissella, Butyrivibrio, Corynebacterium, uncultivated Erysipelotrichaceae, Jeotgallibaca, unclassified Ktedonobacteriaceae, Leptolinea, and uncultivated Ruminiococcaceae. The ability to identify TD risk prior to travel will inform prevention and mitigation strategies to influence diarrhea susceptibility while traveling.

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