Operational Taxonomic Units Composition Research Articles

Woody invaders of a temperate forest have unique root-associated fungal communities

Non-native, invasive plants are increasingly common in Eastern North American forests, but their impact on soil fungi remains unclear. We used DNA metabarcoding to investigate the effects of nativity, plant mycorrhizal type and soil factors on root-associated fungal communities. We focused on understory woody plants in a deciduous forest tract in central New York state, and included three plant types, invasive arbuscular mycorrhizal (AM), native AM, and native ectomycorrhizal (EM), each represented by 3–5 species. We found lower fungal operational taxonomic unit (OTU) diversity in EM than AM plants, but no OTU diversity difference between native and invasive AM species. Pathogen OTU richness and relative abundance were not distinct between plant types. OTU composition was influenced by host mycorrhizal type and by AM plant nativity, with mycorrhizal fungi being important drivers in both cases. The relationships of soil characteristics (e.g., pH) with OTU composition were independent of plant nativity and mycorrhizal type. Specific root length of native AM species was lower than that of invasive AM plants, while EM plants were intermediate. Irrespective of plant type, OTU composition was distinct among plant species, particularly in fungal communities associated with the invader Rhamnus cathartica. These results suggest that invasive AM plants may shift fungal composition relative to native AM and EM plants, with potential long-term consequences for soil biodiversity.

Diversity of ligninolytic ascomycete fungi associated with the bleached leaf litter in subtropical and temperate forests

<p>Little is known regarding the diversity patterns of Xylariaceae and Hypoxylaceae (Ascomycota) fungi taking part in the lignin decomposition of leaf litter from different tree species and under different climatic regions. The alpha and beta diversity of Xylariaceae and Hypoxylaceae fungi was investigated on bleached leaf litter from nine subtropical and cool temperate tree species in Japan. A total of 248 fungal isolates, obtained from 480 leaves from the nine tree species, were classified into 43 operational taxonomic units (OTUs) with a 97% similarity threshold and were assigned to nine genera of Xylariaceae and Hypoxylaceae. There was no overlap of fungal OTUs between subtropical and cool temperate trees. The mean number of fungal OTUs was generally higher in subtropical than cool temperate trees, whereas rarefaction curves depicting the numbers of OTU with respect to the number of leaves from which fungi were isolated were less steep in subtropical trees than in cool temperate trees, reflecting the dominance of major OTUs in the subtropical trees and indicating a higher species richness in cool temperate regions. Nonmetric multidimensional scaling showed general overlaps of fungal OTU compositions among tree species in the respective climatic regions, and one-way permutational multivariate analysis of variance indicated that the OTU composition was not significantly different between the tree species. These results suggest a wide host range and some geographic and climatic structures of distribution of these ligninolytic fungi.</p>

Do aquatic fungal environmental DNA assemblages reflect the surrounding terrestrial sporocarp communities?

The relationship between aquatic environmental DNA (eDNA) assemblages in rivers and the surrounding terrestrial fungal communities has been poorly investigated. Here, we focused on fungi that form soft sporocarps (soft fungi). Two years of sporocarp and aquatic eDNA sampling were conducted at a fragmented forest site, and the soft-fungal assemblages and their temporal dynamics were compared between these two sample types. Aquatic eDNA yielded approximately 1.5 times the operational taxonomic units (OTUs) compared to sporocarps and covered approximately half of the OTUs from sporocarp samples. Lineages that seldom form sporocarps or form inconspicuous sporocarps were successfully detected from aquatic eDNA. Although the OTU composition differed between sporocarp and aquatic eDNA, their temporal dynamics were similar, with both showing a 1-year periodicity. Aquatic eDNA provides insights into fungal diversity and temporal dynamics, but does not fully reflect terrestrial fungi diversity.

The structure of microbial communities in redoximorphic microsites of Gleysol

AbstractMicrobial communities were studied in redoximorphic microsites of highly heterogeneous Gleysol at a mm scale using 16S and 18S amplicon sequencing to demonstrate if the composition of soil microbes reflects the differences in ferric and ferrous micro‐sites. In both explored gley horizons with redoximorphic features (Bg2 and Cg), ferric mottles were significantly enriched with total P and Fe and depleted of O, Si, Al, K and Ca compared with the adjacent ferrous groundmass (SEM–EDS). Ferric mottles were determined as Fe oxide coatings and hypocoatings. In Bg2, both prokaryotic and micro‐eukaryotic communities differed significantly between mottles and groundmass in composition of operational taxonomic units (OTUs) and in proportions of phyla, reflecting heterogeneities in the soil properties there. Mottles in Bg2 were characterized by increased proportion of Proteobacteria, decreased proportion of Acidobacteriota among prokaryotes and by dominance of a single proteobacterial OTU from Anaplasmataceae compared to all other samples. The composition of micro‐eukaryotes showed an opposite trend, as micro‐eukaryotes of Bg2 groundmass were unique among the other horizons, while micro‐eukaryotes of Bg2 mottles had similar composition to neighbouring horizons. Microbial communities of adjacent samples were not more similar to each other than communities of randomly selected ones in Bg2 horizon. That suggests that at mm scale, the sample distance does not represent the driving factor of microbial community composition and that the adjacent samples differ rather due to physicochemical factors. The spatial organization of microbial communities revealed in Bg2 has not reappeared in similarly organized Cg horizon, probably due to other overriding factors. The differences revealed between Bg2 and Cg horizons, including granulometric composition, content of crystalline Fe, exchangeable Al, and organic carbon, as well as exposition to groundwater, were discussed as possible reasons of the distinct organization in Cg. The similarity of pro−/eukaryotic communities of adjacent and non‐adjacent couples suggests no distance decay pattern at a mm scale. The agreement between patchiness in soil properties and microbial communities was revealed for the first time and confirms the importance of microscale patterns in soil.

Community structure and environmental determinants of the bacterial and fungal gut microflora in Hainan gibbons (Nomascus hainanus)

High-throughput sequencing technology was used to establish the OTU (Operational Taxonomic Units) composition and diversity of bacteria and fungi in the gut of three family groups of Hainan gibbons ( Nomascus hainanus ) in the Bawangling National Nature Reserve on Hainan Island, southern China. Firmicutes (47.23%) and Bacteroidetes (36.54%) predominated at the phylum level, while the predominant genera were Prevotella-7 (15.30%), Lachnospiraceae_NK3A20_group (12.49%), and uncultured_bacterium_f_Erysipelotrichaceae (11.79%). At the phylum level, the dominant fungi were Ascomycota (66.41%), as well as Hanseniaspora (23.33%), Schwanniomyces (13.44%), and Pichia (5.43%) at the genus level. We found significant differences in the bacterial OTU diversity (Shannon index) between family groups living in Tropical Lowland or Tropical Montane Rain Forest, compared to those inhabiting Tropical Montane Evergreen Forest at higher altitudes. Bacterial OTU community composition also differed between family groups, unraveling significant differences among the 30 most dominant bacterial core taxa. Furthermore, we unraveled a significant difference of OTU richness in the fungal microbiome (Chao 1) between family groups living at lower altitudes, i.e., Tropical Lowland or Tropical Montane Rain Forest and those inhabiting the Tropical Montane Evergreen Forest. Our analysis further indicated significant differences in the fungal OTU community composition between the three family groups, especially regarding the three most dominant fungal core taxa. Subsequently, two habitat factors, and nine environmental and anthropogenic variables were used to explore possible causes of disparity in the microbial flora of gibbon groups. A factor reduction procedure resulted into three principal components which were correlated to bacterial and fungal OTU richness and diversity using Spearman's rank-order correlations. Bacterial and fungal OTU diversity was high in areas of high altitude, steep slopes, high tree density, but low tree height, while high fungal OTU richness corresponded to high altitude habitats, i.e., in the Tropical Montane Evergreen Forest. Distance to human settlements and to the next water body showed no significant relation with bacterial and fungal richness and diversity.

Desert truffle mycorrhizosphere harbors organic acid releasing plant growth\u2013promoting rhizobacteria, essentially during the truffle fruiting season

Desert truffle is becoming a new crop in semiarid areas. Climatic parameters and the presence of microorganisms influence the host plant physiology and alter desert truffle production. Desert truffle plants present a typical summer deciduous plant phenology divided into four stages: summer dormancy, autumn bud break, winter photosynthetic activity, and spring fruiting. We hypothesize that the bacterial community associated with desert truffle plants will show a seasonal trend linked to their plant growth–promoting rhizobacteria (PGPR) traits. This information will provide us with a better understanding about its potential role in this symbiosis and possible management implementations. Bacteria were isolated from root-adhering soil at the four described seasons. A total of 417 isolated bacteria were phenotypically and biochemically characterized and gathered by molecular analysis into 68 operational taxonomic units (OTUs). They were further characterized for PGPR traits such as indole acetic acid production, siderophore production, calcium phosphate solubilization, and ACCD (1-amino-cyclopropane-1-carboxilatedeaminase) activity. These PGPR traits were used to infer functional PGPR diversity and cultivable bacterial OTU composition at different phenological moments. The different seasons induced shifts in the OTU composition linked to their PGPR traits. Summer was the phenological stage with the lowest microbial diversity and PGPR functions, whereas spring was the most active one. Among the PGPR traits analyzed, P-solubilizing rhizobacteria were harbored in the mycorrhizosphere during desert truffle fruiting in spring.

Intestinal Tract Microbe Communities Associated with Horseshoe Crabs from Beibu Gulf, China.

Until now, there has been little research on the intestinal microbial community of horseshoe crabs. To fill this gap, we investigated the microbiome composition of the Chinese horseshoe crab, Tachypleus tridentatus, and the mangrove horseshoe crab, Carcinoscorpius rotundicauda. We sequenced the 16S rRNA gene of intestinal bacterial species and compared the microbial community structure and diversity. Next, we show that the total effective bacterial sequence was 36,865 reads, and the average annotated operational taxonomic unit (OTU) number was 240. Through hierarchical clustering analysis and principal coordinate analysis samples from two horseshoe crab species, we found that the intestinal flora of the same horseshoe crab species was relatively concentrated, while the microbiome of a different horseshoe crab species were significantly separated. Cluster analysis showed that two samples, one from Chinese horseshoe crabs and one from mangrove horseshoe crabs, had similar microbial community structure, while other samples were relatively discrete. The gut microbiota of the mangrove horseshoe crab were dominated by the phyla Tenericutes (42.71%), Firmicutes (24.27%), and Proteobacteria (20.39%), while the top three phyla in the Chinese horseshoe crab intestinal tract were Tenericutes (57.19%), Proteobacteria (22.14%), and Bacteroidetes (7.38%). To intuitively understand the similarity and overlap of the OTU composition of each group, we performed Venn diagram analysis. The two species shared 284 OTUs, accounting for 81.8% of the total. This indicates that although there is high similarity between mangrove and Chinese horseshoe crab in gastrointestinal microbial community structure, there are also some differences, which deserve further discussion.

Diversity analysis of gut microbiota between healthy controls and those with atopic dermatitis in a Chinese population.

An increasing body of evidence suggests that gut microbiota is involved in atopic dermatitis (AD). We aimed to use high-throughput sequencing to characterize the differences in the composition of the gut microbiota between healthy controls and patients with AD. Fecal samples from 93 volunteers were analyzed using 16S rRNA sequencing, including 44 patients with AD and 49 healthy control subjects, aged 6-22years. Our data show that the operational taxonomic unit composition in patients with AD had greater component similarity than the healthy controls. Patients with AD had a lower alpha diversity than healthy control subjects. The relative abundance of Porphyromonadaceae, Blautia, Parabacteroides, Bacteroides ovatus, Bacteroides uniformis and Prevotella stercorea was significantly higher (P<0.05) in patients with AD than healthy control subjects. Clostridium and P.stercorea were higher (P<0.05) in healthy control subjects compared with patients with AD. The results of linear discriminant analysis effect size show that Bacteroidaceae and Porphyromonadaceae can act as possible biomarkers associated with diagnosis of AD. However, this needs further experimental verification. Taken together, these results demonstrate the changes in microbiota composition in AD compared with a healthy control group, opening the way to future diagnosis or intervention studies.

Can metabarcoding resolve intraspecific genetic diversity changes to environmental stressors? A test case using river macrozoobenthos

Genetic diversity is the most basal level of biodiversity and determines the evolutionary capacity of species to adapt to changing environments, yet it is typically neglected in routine biomonitoring and stressor impact assessment. For a comprehensive analysis of stressor impacts on genetic diversity, it is necessary to assess genetic variants simultaneously in many individuals and species. Such an assessment is not as straightforward and usually limited to one or few focal species. However, nowadays species diversity can be assessed by analysing thousands of individuals of a community simultaneously with DNA metabarcoding. Recent bioinformatic advances also allow for the extraction of exact sequence variants (ESVs or haplotypes) in addition to Operational Taxonomic Units (OTUs). By using this new capability, we here evaluated if the analysis of intraspecific mitochondrial diversity in addition to species diversity can provide insights into responses of stream macrozoobenthic communities to environmental stressors. For this purpose, we analysed macroinvertebrate bulk samples of three German river systems with different stressor levels using DNA metabarcoding. While OTU and haplotype number were negatively correlated with stressor impact, this association was not as clear when studying haplotype diversity across all taxa. However, stressor responses were found for sensitive EPT (Ephemeroptera, Plecoptera, Trichoptera) taxa and those exceedingly resistant to organic stress. An increase in haplotype number per OTU and haplotype diversity of sensitive taxa was observed with an increase in ecosystem quality and stability, while the opposite pattern was detected for pollution resistant taxa. However, this pattern was less prominent than expected based on the strong differences in stressor intensity between sites. To compare genetic diversity among communities in river systems, we focussed on OTUs, which were present in all systems. As OTU composition differed strongly between rivers, this led to the exclusion of a high number of OTUs, especially in diverse river systems of good quality, which potentially diminished the increase in intraspecific diversity. To better understand responses of intraspecific genetic diversity to environmental stressors, for example in river ecosystems, it would be important to increase OTU overlap between compared sites, e.g. by sampling a narrower stressor gradient, and to perform calibrated studies controlling for the number of individuals and their haplotypes. However, this pioneer study shows that the extraction of haplotypes from DNA metabarcoding datasets is a promising source of information to simultaneously assess intraspecific diversity changes in response to environmental impacts for a metacommunity.

Effect of floristic composition and configuration on plant root mycobiota: a landscape transposition at a small scale.

Fungal communities in the root endosphere are heterogeneous at fine scale. The passenger hypothesis assumes that this heterogeneity is driven by host plant distribution. Plant composition and host plant configuration should then influence root fungal assemblages. We used a large-scale experimental design of 25 mixtures of grassland plants. We sampled Brachypodium pinnatum in each mesocosm, and used amplicon mass-sequencing to analyze the endospheric mycobiota. We used plant distribution maps to assess plant species richness and evenness (heterogeneity of composition), and patch size and the degree of isolation of B.pinnatum (heterogeneity of configuration) on fungal community assembly. The Glomeromycotina community in B.pinnatum roots was not related to either floristic heterogeneity or productivity. For Ascomycota, the composition of operational taxonomic units (OTUs) was driven by plant evenness while OTU richness decreased with plant richness. For Basidiomycota, richness increased with host plant aggregation and connectivity. Plant productivity influenced Ascomycota, inducing a shift in OTU composition and decreasing evenness. Plant heterogeneity modified root mycobiota, with potential direct (i.e. host preference) and indirect (i.e. adaptations to abiotic conditions driven by plant occurrence over time) effects. Plant communities can be envisioned as microlandscapes consisting of a variety of fungal niches.

Characterization and comprehensive analysis of the ecological interaction networks of bacterial communities in Paullinia cupana var. sorbilis by 16S rRNA gene metabarcoding.

Endophytes improve the host performance in areas of high plant endemicity. Paullinia cupana is an Amazonia plant species of economic and social importance due to the high caffeine concentration in its seeds. An interesting strategy to identify endophytic microorganisms with potential biotechnological application is to understand the factors that influence the endophytic community to rationalize the host management programs. We used the next-generation sequencing for bacterial 16S rRNA gene to examine how the P. cupana organ, genotype, and geographic location influenced its endophytic bacterial community. We obtained 1520 operational taxonomic units (OTUs) distributed in 19 phyla, 32 classes, 79 orders, 114 families and 174 genera. The P. cupana roots and leaves were specifically colonized by the bacterial genera Acidothermus and Porphyromonas, respectively, with high relative frequency. The plant organ type influenced the endophytic community's richness, diversity, OTUs composition, relative abundance of phyla and genera, and genera interaction network. However, the host plant genotype and geographic location influenced the composition and interaction among genera in the network analysis. Prevotella is a super-generalist genus in the interaction network of endophytic bacteria of P. cupana. This study revealed endophytic bacterial groups of importance to P. cupana and stressed that the host plant organ modulates the structure and interactions within this community. Our results indicated that the microbial community adapted to colonize P. cupana by adjusting to its composition and interaction network. The isolation of abundant and super-generalist bacterial genera shall help to examine their functionality to the composition and fitness of the endophytic community of P. cupana.

Mangrove-diazotroph relationships at the root, tree and forest scales: diazotrophic communities create high soil nitrogenase activities in Rhizophora stylosa rhizospheres.

The tidal flats on which mangrove plants grow tend to have low soil nitrogen contents because nitrogen-containing litter is repeatedly washed offshore by ebb tides. Under such circumstances, it is unclear how mangrove plants acquire the nitrogen required to support their vigorous growth. In the present work, chemical and biological characteristics of diazotrophy around mangrove plant roots were surveyed under natural conditions to elucidate mangrove-diazotroph relationships. Soil nitrogenase activity of a representative mangrove plant, Rhizophora stylosa, which has a broad geographical distribution, was measured using the acetylene reduction assay at forest, tree and prop root scales. In addition, diazotrophic community composition was compared between rhizosphere and bulk soil based on sequencing of nifH genes. Soil nitrogenase activity was high near prop roots, and this pattern was enhanced as soil live root content increased. At the forest scale, we observed high soil nitrogenase activity (acetylene-reducing activity) inside the forest (the highest value was 90.9 µmol C2H2 min-1 cm-3, average 46.8 ± 18.2 µmol C2H2 min-1 cm-3). Rates decreased sharply from the forest to the tidal flat (range 1.2-22.2 µmol C2H2 min-1 cm-3, average 7.9 ± 4.5 µmol C2H2 min-1 cm-3). The nifH operational taxonomic unit composition differed significantly among forest and tree rhizospheres and the bulk soil (P < 0.0001). Our results suggest that the accumulation of diazotrophs around R. stylosa mangrove trees enhances the supply of biologically fixed nitrogen to the mangrove roots. This supply is especially important when the soil naturally contains little nitrogen. This nitrogen acquisition system may be a key process that explains the high productivity of mangrove ecosystems.

Irradiation effect on the structure of bacterial communities associated with the oriental fruit fly, Bactrocera dorsalis

AbstractThe role of symbiotic microbes in insects, especially the beneficial character of this interaction for insects, has received much attention in recent years as it has been related to important aspects of the host insects' biology such as development, reproduction, survival, and fitness. Among insect hosts, tephritid fruit flies are well known to form beneficial associations with their symbionts. To control these destructive agricultural pests, environmentally friendly approaches, like the sterile insect technique as a component of integrated pest management strategies, remain most effective. In this study, changes in the bacterial profile of mass‐reared oriental fruit flies, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), were examined in both larval and adult stages and also after irradiation by employing a 16S rRNA gene‐based Illumina sequencing approach. Proteobacteria was the prevalent bacterial phylum in non‐irradiated adults and larvae. Alphaproteobacteria was the most abundant class in larvae but almost absent in adults, which was dominated by Gammaproteobacteria. Firmicutes were present in both developmental stages but at lower relative abundance. At genus level, Acetobacter prevailed in the larval stage and members of the Enterobacteriaceae family in adults. Irradiated samples exhibited higher diversity and richness indices compared to the non‐irradiated oriental fruit flies, whereas no significant changes were observed between the two developmental stages of the non‐irradiated samples. Lactobacillus, members of the Orbacecae family, and Morganella were detected but to a lesser degree upon irradiation, whereas the relative abundance of Lactococcus and Orbus increased. The bacterial profile of larvae appeared to be different compared to that of adult B. dorsalis flies. The subsequent application of irradiation at the pupal stage led to the development of different microbiota between treated and untreated samples, affecting diversity and operational taxonomic unit composition. Irradiated samples of oriental fruit flies were characterized by higher species diversity and richness.

Potential role of host-derived quorum quenching in modulating bacterial colonization in the moon jellyfish Aurelia aurita

Multicellular organisms can be regarded as metaorganisms, comprising of a macroscopic host interacting with associated microorganisms. Within this alliance, the host has to ensure attracting beneficial bacteria and defending against pathogens to establish and maintain a healthy homeostasis. Here, we obtained several lines of evidence arguing that Aurelia aurita uses interference with bacterial quorum sensing (QS) - quorum quenching (QQ) - as one host defense mechanism. Three A. aurita-derived proteins interfering with bacterial QS were identified by functionally screening a metagenomic library constructed from medusa-derived mucus. Native expression patterns of these host open reading frames (ORFs) differed in the diverse life stages (associated with different microbiota) pointing to a specific role in establishing the developmental stage-specific microbiota. Highly increased expression of all QQ-ORFs in germ-free animals further indicates their impact on the microbiota. Moreover, incubation of native animals with pathogenic bacteria induced expression of the identified QQ-ORFs arguing for a host defense strategy against confronting bacteria by interference with bacterial QS. In agreement, immobilized recombinant QQ proteins induced restructuring of polyp-associated microbiota through changing abundance and operational taxonomic unit composition. Thus, we hypothesize that additional to the immune system host-derived QQ-activities potentially control bacterial colonization.

Gcd Gene Diversity of Quinoprotein Glucose Dehydrogenase in the Sediment of Sancha Lake and Its Response to the Environment.

Quinoprotein glucose dehydrogenase (GDH) is the most important enzyme of inorganic phosphorus-dissolving metabolism, catalyzing the oxidation of glucose to gluconic acid. The insoluble phosphate in the sediment is converted into soluble phosphate, facilitating mass reproduction of algae. Therefore, studying the diversity of gcd genes which encode GDH is beneficial to reveal the microbial group that has a significant influence on the eutrophication of water. Taking the eutrophic Sancha Lake sediments as the research object, we acquired samples from six sites in the spring and autumn. A total of 219,778 high-quality sequences were obtained by DNA extraction of microbial groups in sediments, PCR amplification of the gcd gene, and high-throughput sequencing. Six phyla, nine classes, 15 orders, 29 families, 46 genera, and 610 operational taxonomic units (OTUs) were determined, suggesting the high genetic diversity of gcd. Gcd genes came mainly from the genera of Rhizobium (1.63–77.99%), Ensifer (0.13–56.95%), Shinella (0.32–25.49%), and Sinorhizobium (0.16–11.88%) in the phylum of Proteobacteria (25.10–98.85%). The abundance of these dominant gcd-harboring bacteria was higher in the spring than in autumn, suggesting that they have an important effect on the eutrophication of the Sancha Lake. The alpha and beta diversity of gcd genes presented spatial and temporal differences due to different sampling site types and sampling seasons. Pearson correlation analysis and canonical correlation analysis (CCA) showed that the diversity and abundance of gcd genes were significantly correlated with environmental factors such as dissolved oxygen (DO), phosphorus hydrochloride (HCl–P), and dissolved total phosphorus (DTP). OTU composition was significantly correlated with DO, total organic carbon (TOC), and DTP. GDH encoded by gcd genes transformed insoluble phosphate into dissolved phosphate, resulting in the eutrophication of Sancha Lake. The results suggest that gcd genes encoding GDH may play an important role in lake eutrophication.

Rice straw biochar as a novel niche for improved alterations to the cecal microbial community in rats

Biochar as additive has been shown positive effect in animal production, which may be linked to the role of gastrointestinal microbial modulation. This study aimed to assess the effects of biochar on the gut microbial communities in terms of their structure and diversity. Illumina high-throughput technology was utilized to evaluate the cecal microbial community in Wistar rats received oral rice straw biochar (RSB) at 1120 mg/kg of body weight for 5 weeks. RSB improved the gut mucosal structure and epithelial integrity. More importantly, principal coordinate analysis of UniFrac distances based on a 97% operational taxonomic unit composition and abundance indicated that the bacterial community was ameliorated after RSB addition (P < 0.05). Firmicutes and Bacteroidetes were found to be the prevalent phyla accounting for approximately 90% of the sequences and their ratio of relative abundance was increased by RSB addition (P < 0.05). Improved bacterial proportion of unclassified Lachnospiraceae (P < 0.001), Oscillibacter (P = 0.02), and Clostridium IV (P = 0.02) and XIVa (P = 0.02) as well as decreased abundances of Prevotella (P < 0.001) and Bacteroides (P = 0.03) were also detected at genus level following RSB treatment. These results revealed that RSB altered and improved the cecal microbial community, which may contribute to the affected growth and gut status in rats.

Ecological patterns in the skin microbiota of frogs from tropical Australia.

The microbiota of frog skin can play an important role in protecting against diseases and parasites. The frog skin microbial community represents a complex mix of microbes that are promoted by the chemical environment of the frog skin and influenced by the animal's immediate past environment. The microbial communities of six species of frogs sampled from the campus of Charles Darwin University (CDU) were more similar within species than between species. The microbiota of the introduced cane toad (Rhinella marina) was most dissimilar among the species. Pairwise comparisons showed that the microbial communities of each species were different, except for the terrestrial Litoria nasuta and the arboreal L. rothii. The microbial communities of the six species were not related to ecological habit (arboreal or terrestrial), and neither was the alpha diversity of the microbes. The core microbes (defined as being on ≥90% of individuals of a species or group) were significantly different among all species, although 89 microbial operational taxonomic units (OTUs) were core microbes for all six species at CDU. Two species, Rhinella marina and Litoria rothii, were sampled at additional sites approximately 10 and 30 km from CDU. The microbial communities and the core OTU composition were different among the sites, but there were nevertheless 194 (R. marina) and 181 (L. rothii) core OTUs present at all three sites. Thus, the core microbiota varied with respect to geographic range and sample size.

Compositional analysis of archaeal communities in high and low microbial abundance sponges in the Misool coral reef system, Indonesia

ABSTRACTThe high/low microbial abundance (HMA/LMA) dichotomy in sponges has been the subject of several studies over recent years, but few studies have analysed this dichotomy in terms of the sponge archaeal community and function. Using a 16S rRNA gene barcoded pyrosequencing approach and predictive functional analysis (PICRUSt) we compared the archaeal composition, richness and predicted function of one HMA sponge (Xestospongia testudinaria), one LMA sponge (Stylissa carteri) and one sponge species of unknown microbial abundance (Aaptos lobata). Although most of the archaeal sequences were assigned to the Crenarchaeota phylum, S. carteri had the highest percentage of sequences assigned to the Euryarchaeota phylum. Variation among sponge species explained >85% of the variation in archaeal operational taxonomic unit (OTU) composition with each sponge species forming a distinct cluster. There were significant differences in predicted PICRUSt profiles among sponge species, suggesting that archaeal communities present in the studied sponge species may perform different functions. X. testudinaria and A. lobata were similar both in terms of OTU and KEGG orthologues composition, which may indicate that A. lobata is a HMA sponge. Additionally, some of the most enriched functions seem to be related to traits associated with high and low microbial abundance sponges.

Differences in structure of northern Australian hypolithic communities according to location, rock type, and gross morphology.

Hypolithic communities (under translucent rocks) were compared between a semi-arid site (Wave Hill) and a site with considerably higher rainfall (Lake Bennett) to test the hypothesis that the communities at the higher rainfall site would be more diverse. A total of 153 cyanobacteria operational taxonomic units (OTUs) were identified, and only 50 of those were found at both sites. Of these, only two were core OTUs, as defined as being present in ≥90% of samples, highlighting the extreme differences in the cyanobacterial communities at the two sites. At Wave Hill, we compared the composition of the cyanobacterial components under two different rock types (quartz and prehnite) to determine if the different minerals would result in different hypolithic communities, but no differences were found. Of the 42 core OTUs found at Wave Hill, 22 (52%) were shared between the two rock types. As hypothesised, the diversity of both cyanobacteria and eukaryotes in the hypolithic communities was significantly higher at Lake Bennett. Some hypolithic communities were thin and tightly adhered to the rock surface, but others were thicker and could be peeled off the rock in sheets. However, the two types were not significantly different in OTU composition. Metazoans, primarily nematodes, were ubiquitous, raising the possibility that nematodes may act as vectors to transport the components of hypolithic communities from rock to rock as a mechanism of colonization.

Broccoli Consumption Impacts the Human Gastrointestinal Microbiota

BackgroundThe human gastrointestinal (GI) microbiota is increasingly linked to health outcomes; however, our understanding of how specific foods alter the microbiota is limited. Cruciferous vegetables such as broccoli have been associated with cancer prevention as a result of their high levels of glucosinolates, especially glucoraphanin. The activation of glucoraphanin to its isothiocyanate form, sulforaphane, is dependent on the enzyme myrosinase. Myrosinase is naturally found in broccoli, but is degraded by cooking. It has been hypothesized that plasma sulforaphane observed in persons eating cooked broccoli may be related to glucoraphanin hydrolysis by microbial myrosinase. Myrosinase activity is greater in rodents who regularly consume cruciferous vegetables compared to those who do not, suggesting a priming effect on the microbiota. However, it remains to be discovered which members of the microbiota are responsible for this action in clinical populations.ObjectiveWe aimed to determine the impact of broccoli consumption on the GI microbiota of healthy adult men and women as a means to understand the role of microbes in glucosinolate hydrolysis.MethodsA controlled feeding, randomized, crossover study consisting of two 17‐day treatment periods separated by a 24‐day washout was conducted in healthy adults (n=18). Participants were fed at weight maintenance, and received a Brassica‐free control diet or the same diet with 200 g of cooked broccoli and 20 g of fresh daikon radish (as a source of plant myrosinase) per day. Fecal samples were collected at baseline and at the end of each treatment period for microbial analysis. Following DNA isolation, bacterial, archaeal, and fungal barcoded amplicon pools were generated using a Fluidigm Access Array system prior to high‐throughput sequencing on an Illumina MiSeq. Data were analyzed using QIIME 1.8.0 and 1.9.1 and SAS 9.4.ResultsBeta diversity analysis based on 97% operational taxonomic unit composition and abundances indicated that bacterial communities were impacted by treatment (p=0.03). Broccoli consumption resulted in a significant change in the proportion of Bacteroidetes relative to Firmicutes, whereby participants consuming broccoli increased this ratio by 37% from baseline to end, compared to a 5% reduction during the control period (p=0.02). Bacteroides increased by 6% following broccoli consumption and decreased by 2% in the control group, but the difference between these two groups was not statistically significant (p=0.15).ConclusionThese novel results reveal that broccoli consumption affects the diversity and composition of the GI microbiota of healthy adults. These data help fill the gap in knowledge related to the role of bacterial hydrolysis of phytonutrients. The increase in Bacteroides spp. is particularly relevant because Bacteroides thetaiotaomicron has been shown in vitro to utilize glucosinolates. Additional study is ongoing to determine if phenotypic responses are related to archaea and body mass index.Support or Funding InformationFunding provided by the National Cancer Institute and the USDA.