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Is there convergence of gut microbes in blood-feeding vertebrates?

Animal microbiomes play an important role in dietary adaptation, yet the extent to which microbiome changes exhibit parallel evolution is unclear. Of particular interest is an adaptation to extreme diets, such as blood, which poses special challenges in its content of proteins and lack of essential nutrients. In this study, we assessed taxonomic signatures (by 16S rRNA amplicon profiling) and potential functional signatures (inferred by Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt)) of haematophagy in birds and bats. Our goal was to test three alternative hypotheses: no convergence of microbiomes, convergence in taxonomy and convergence in function. We find a statistically significant effect of haematophagy in terms of microbial taxonomic convergence across the blood-feeding bats and birds, although this effect is small compared to the differences found between haematophagous and non-haematophagous species within the two host clades. We also find some evidence of convergence at the predicted functional level, although it is possible that the lack of metagenomic data and the poor representation of microbial lineages adapted to haematophagy in genome databases limit the power of this approach. The results provide a paradigm for exploring convergent microbiome evolution replicated with independent contrasts in different host lineages.

Philosophical Transactions of the Royal Society B:

Plant diversity predicts beta but not alpha diversity of soil microbes across grasslands worldwide

Aboveground–belowground interactions exert critical controls on the composition and function of terrestrial ecosystems, yet the fundamental relationships between plant diversity and soil microbial diversity remain elusive. Theory predicts predominantly positive associations but tests within single sites have shown variable relationships, and associations between plant and microbial diversity across broad spatial scales remain largely unexplored. We compared the diversity of plant, bacterial, archaeal and fungal communities in one hundred and forty-five 1 m 2 plots across 25 temperate grassland sites from four continents. Across sites, the plant alpha diversity patterns were poorly related to those observed for any soil microbial group. However, plant beta diversity (compositional dissimilarity between sites) was significantly correlated with the beta diversity of bacterial and fungal communities, even after controlling for environmental factors. Thus, across a global range of temperate grasslands, plant diversity can predict patterns in the composition of soil microbial communities, but not patterns in alpha diversity.

Ecology Letters

Preliminary geochemical, microbiological, and epidemiological investigations into possible linkages between lignite aquifers, pathogenic microbes, and kidney disease in northwestern Louisiana

In May 2002, 15 wells and four surface water sites were sampled, and in September 2002, those same wells and sites plus four additional surface sites were sampled in five parishes of northwestern Louisiana. A geographic information system (GIS) was used to select residential water wells for sampling. Well water samples were analyzed for pH, conductivity, organic compounds, and nutrient and anion concentrations. All samples were further tested for presence of fungi (maintained for up to 28 days and colonies counted and identified microscopically), and metal and trace element concentration by inductively-coupled plasma mass spectrometry and atomic emission spectrometry. Surface water samples were tested for dissolved oxygen and evidence of leptospiral bacterial presence. A polymerase chain reaction protocol was optimized for detection of pathogenic leptospires, and the sensitivity of the assay was determined. The Spearman correlation method was used to assess the association between the endpoints for these field/laboratory analyses and the incidence of cancer of the renal pelvis obtained from the Louisiana Tumor Registry. Significant associations were revealed between the cancer rate and the overall number of organic compounds, the fungi Zygomycetes, the nutrients PO4 and NH3, and thirteen chemical elements (As, B, Br, Cl, Cr, F, Li, Na, P, Rb, Se, Sr, W) from the well water as compared to the controls. Among the species of fungi from the total of 136 isolates were 12 Penicillium spp., at least two Aspergillus spp., a number of other genera (Alternaria sp., Eupenicillium lapidosum, Cladosporium sp., Epicoccum sp., Trichoderma sp., Paecilomyces sp., Chrysosporium sp., Chloridium sp.), and Zygomycetes, and Coelmycetes -- some of which are known mycotoxin producers. The two control wells yielded a mean of 6.5 (SD = 3.5355) individual isolates, while the mean number of isolates from all other sites was 7.6 (SD = 4.4866). Presence of human pathogenic leptospires was detected in 4/8 (50 percent) of the surface water sites sampled. These initial results suggest that additional investigation into these relationships is warranted.

Open-File Report

Endophytic bacteria in grass crop growth promotion and biostimulation

Plants naturally carry microbes on seeds and within seeds that may facilitate development and early survival of seedlings. Some crops have lost seed-vectored microbes in the process of domestication or during seed storage and seed treatment. Biostimulant microbes from wild plants were used by pre-modern cultures to re-acquire beneficial seed microbes. Today some companies have developed or are developing the use of microbes obtained from soils or plant sources to stimulate plant development and growth. Many of these biostimulant microbes are endophytic in plants. Biostimulant products also include humic substances, which appear to function as signal molecules in plants, triggering increased internalization of soil microbes into root cells and tissues. In addition, protein coatings on seeds fuel the growth of seed surface-vectored microbes, increasing microbial activity around and within roots. In this article, we provide evidence of the endophytic nature of many biostimulant microbes, and suggest that many of the beneficial effects of microbial biostimulants stem from their action as endophytes or as participants or stimulants of rhizophagy cycle activity.

Grass Research

Lack of surface-associated microorganisms in a mixed species community of freshwater Unionidae

To determine whether unionids contain surface-attached endosymbiotic bacteria, ciliates, or fungi, we used scanning electron microscopy to examine the epithelial surface of various organs within the digestive systems and mantle cavity of temperate river and lake unionids on a seasonal basis. We also cultured material removed from the lumen of these same organs and from the mantle cavity to detect cellobiose-, cellulose-, and chitin- degrading microbes. No true endosymbiotic fauna were observed attached to the surface of the digestive or mantle tissues of any species of unionid. Microbial growth on cellulose or chitin bacteriological media varied by season and habitat, but not by unionid species or source of the isolate. Lake unionids did not contain microbes capable of digesting cellulose or chitin, whereas unionids from the river site did in March and August, but not in December. Since these cultured cellulose- and chitin-degrading bacteria were never found attached to any unionid tissues, they appeared to be transient forms, not true endosymbionts. Microbes capable of digesting cellobiose were found in every unionid collected in all seasons and habitats, but again, no microbes were directly observed attached to unionid tissues. If unionids, like most other vertebrates, lack digestive enzymes required to initiate primary bond breakage, then the lack of cellulolytic and chitinolytic endosymbionts would affect the ability to utilize cellulose or chitin foods. Thus, in captivity dry feeds based on corn, soybeans, or nauplii should be pre-digested to ensure maximum absorption of nutrients by unionids. The lack of celluloytic or chitinolytic endosymbionts should not affect relocation success, though the seasonal role of transient microbes in unionid nutrition requires further investigation.

Journal of Shellfish Research

Acetylenotrophic and diazotrophic Bradyrhizobium sp. strain I71 from TCE-contaminated soils

Abstract Acetylene (C 2 H 2 ) is a molecule rarely found in nature, with very few known natural sources, but acetylenotrophic microorganisms can use acetylene as their primary carbon and energy source. As of 2018 there were 15 known strains of aerobic and anaerobic acetylenotrophs; however, we hypothesize there may yet be unrecognized diversity of acetylenotrophs in nature. This study expands the known diversity of acetylenotrophs by isolating the aerobic acetylenotroph, Bradyrhizobium sp. strain I71, from trichloroethylene (TCE)-contaminated soils. Strain I71 is a member of the class Alphaproteobacteria and exhibits acetylenotrophic and diazotrophic activities, the only two enzymatic reactions known to transform acetylene. This unique capability in the isolated strain may increase the genus’ economic impact beyond agriculture as acetylenotrophy is closely linked to bioremediation of chlorinated contaminants. Computational analyses indicate that the Bradyrhizobium sp. strain I71 genome contains 522 unique genes compared to close relatives. Moreover, applying a novel hidden Markov model of known acetylene hydratase (AH) enzymes identified a putative AH enzyme. Protein annotation with I-TASSER software predicted the AH from the microbe Syntrophotalea acetylenica as the closest structural and functional analog. Furthermore, the putative AH was flanked by horizontal gene transfer (HGT) elements, like that of AH in anaerobic acetylenotrophs, suggesting an unknown source of acetylene or acetylenic substrate in the environment that is selecting for the presence of AH. Importance The isolation of Bradyrhizobium strain I71 expands the distribution of acetylene-consuming microbes to include a group of economically important microorganisms. Members of Bradyrhizobium are well studied for their abilities to improve plant health and increase crop yields by providing bioavailable nitrogen. Additionally, acetylene-consuming microbes have been shown to work in tandem with other microbes to degrade soil contaminants. Based on genome, cultivation, and protein prediction analysis, the ability to consume acetylene is likely not widespread within the genus Bradyrhizobium . These findings suggest that the suite of phenotypic capabilities of strain I71 may be unique and make it a good candidate for further study in several research avenues.

California

Root endophytes and invasiveness: no difference between native and non‐native Phragmites in the Great Lakes Region

Microbial interactions could play an important role in plant invasions. If invasive plants associate with relatively more mutualists or fewer pathogens than their native counterparts, then microbial communities could foster plant invasiveness. Studies examining the effects of microbes on invasive plants commonly focus on a single microbial group (e.g., bacteria) or measure only plant response to microbes, not documenting the specific taxa associating with invaders. We surveyed root microbial communities associated with co‐occurring native and non‐native lineages of Phragmites australis, across Michigan, USA. Our aim was to determine whether (1) plant lineage was a stronger predictor of root microbial community composition than environmental variables and (2) the non‐native lineage associated with more mutualistic and/or fewer pathogenic microbes than the native lineage. We used microscopy and culture‐independent molecular methods to examine fungal colonization rate and community composition in three major microbial groups (bacteria, fungi, and oomycetes) within roots. We also used microbial functional databases to assess putative functions of the observed microbial taxa. While fungal colonization of roots was significantly higher in non‐native Phragmites than the native lineage, we found no differences in root microbial community composition or potential function between the two Phragmites lineages. Community composition did differ significantly by site, with soil saturation playing a significant role in structuring communities in all three microbial groups. The relative abundance of some specific bacterial taxa did differ between Phragmites lineages at the phylum and genus level (e.g., Proteobacteria, Firmicutes ). Purported function of root fungi and respiratory mode of root bacteria also did not differ between native and non‐native Phragmites . We found no evidence that native and non‐native Phragmites harbored distinct root microbial communities; nor did those communities differ functionally. Therefore, if the trends revealed at our sites are widespread, it is unlikely that total root microbial communities are driving invasion by non‐native Phragmites plants.

Ecosphere

Fire modifies plant–soil feedbacks

Although plant–soil feedbacks (interactions between plants and soils, often mediated by soil microbes, abbreviated as PSFs) are widely known to influence patterns of plant diversity at local and landscape scales, these interactions are rarely examined in the context of important environmental factors. Resolving the roles of environmental factors is important because the environmental context may alter PSF patterns by modifying the strength or even direction of PSFs for certain species. One important environmental factor that is increasing in scale and frequency with climate change is fire, though the influence of fire on PSFs remains essentially unexamined. By changing microbial community composition, fire may alter the microbes available to colonize the roots of plants and thus seedling growth post-fire. This has potential to change the strength and/or direction of PSFs, depending on how such changes in microbial community composition occur and the plant species with which the microbes interact. We examined how a recent fire altered PSFs of two leguminous, nitrogen-fixing tree species in Hawaiʻi. For both species, growing in conspecific soil resulted in higher plant performance (as measured by biomass production) than growing in heterospecific soil. This pattern was mediated by nodule formation, an important process for growth for legume species. Fire weakened PSFs for these species and therefore pairwise PSFs, which were significant in unburned soils, but were nonsignificant in burned soils. Theory suggests that positive PSFs such as those found in unburned sites would reinforce the dominance of species where they are locally dominant. The change in pairwise PSFs with burn status shows PSF-mediated dominance might diminish after fire. Our results demonstrate that fire can modify PSFs by weakening the legume-rhizobia symbiosis, which may alter local competitive dynamics between two canopy dominant tree species. These findings illustrate the importance of considering environmental context when evaluating the role of PSFs for plants.

Ecology

Captivity, reintroductions, and the rewilding of amphibian-associated bacterial communities

Many studies have noted differences in microbes associated with animals reared in captivity compared to their wild counterparts, but few studies have examined how microbes change when animals are reintroduced to the wild after captive rearing. As captive assurance populations and reintroduction programs increase, a better understanding of how microbial symbionts respond during animal translocations is critical. We examined changes in microbes associated with boreal toads ( Anaxyrus boreas ), a threatened amphibian, after reintroduction to the wild following captive rearing. Previous studies demonstrate that developmental life stage is an important factor in amphibian microbiomes. We collected 16S marker-gene sequencing datasets to investigate: (i) comparisons of the skin, mouth, and fecal bacteria of boreal toads across four developmental life stages in captivity and the wild, (ii) tadpole skin bacteria before and after reintroduction to the wild, and (iii) adult skin bacteria during reintroduction to the wild. We demonstrated that differences occur across skin, fecal, and mouth bacterial communities in captive versus wild boreal toads, and that the degree of difference depends on developmental stage. Skin bacterial communities from captive tadpoles were more similar to their wild counterparts than captive post-metamorphic individuals were to their wild counterparts. When captive-reared tadpoles were introduced to a wild site, their skin bacteria changed rapidly to resemble wild tadpoles. Similarly, the skin bacterial communities of reintroduced adult boreal toads also shifted to resemble those of wild toads. Our results indicate that a clear microbial signature of captivity in amphibians does not persist after release into natural habitat.

Colorado

Evaluation of the functional roles of fungal endophytes of Phragmites australis from high saline and low saline habitats

Non-native Phragmites australis decreases biodiversity and produces dense stands in North America. We surveyed the endophyte communities in the stems, leaves and roots of collections of P. australis obtained from two sites with a low and high salt concentration to determine differences in endophyte composition and assess differences in functional roles of microbes in plants from both sites. We found differences in the abundance, richness and diversity of endophytes between the low saline collections (18 species distributed in phyla Ascomycota, Basidiomycota and Stramenopiles (Oomycota); from orders Dothideales, Pleosporales, Hypocreales, Eurotiales, Cantharellales and Pythiales; Shannon H = 2.639; Fisher alpha = 7.335) and high saline collections (15 species from phylum Ascomycota; belonging to orders Pleosporales, Hypocreales, Diaporthales, Xylariales and Dothideales; Shannon H = 2.289; Fisher alpha = 4.181). Peyronellaea glomerata , Phoma macrostoma and Alternaria tenuissima were species obtained from both sites. The high salt endophyte community showed higher resistance to zinc, mercury and salt stress compared to fungal species from the low salt site. These endophytes also showed a greater propensity for growth promotion of rice seedlings (a model species) under salt stress. The results of this study are consistent with the ‘habitat-adapted symbiosis hypothesis’ that holds that endophytic microbes may help plants adapt to extreme habitats. The capacity of P. australis to establish symbiotic relationships with diverse endophytic microbes that enhance its tolerance to abiotic stresses could be a factor that contributes to its invasiveness in saline environments. Targeting the symbiotic associates of P. australis could lead to more sustainable control of non-native P. australis .

Biological Invasions

Influence of soil microbiota on Taxodium distichum seedling performance during extreme flooding events

Plant associations with soil microbiota can modulate tree seedling growth and survival via mutualistic or antagonistic interactions. It is uncertain, however, whether soil microbiota influence seedling growth of coastal trees when exposed to extreme flooding regimes. We evaluated the role of soil microbes in promoting baldcypress ( Taxodium distichum ) seedling performance under different inundation scenarios and determined the influence of flooding on the colonization of in planta beneficial microbes. Seedlings reared in sterile and non-sterile soil were exposed to three different flooding regimes historically experienced in Louisiana swamps. Seedling growth was assessed, and the colonization by beneficial symbionts such as arbuscular mycorrizhal fungi (AMF), and dark septate endophytes (DSE) was evaluated in harvested roots. Seedlings grown in sterile soil had six times higher growth than seedlings reared in non-sterile soil. As a result, we evaluated pathogen load in the roots by assessing oomycete colonization. Flooding influenced the in planta colonization of DSE and oomycetes, but did not affect the colonization of mutualist AMF fungi. DSE and oomycetes were rarer in flooded conditions, while AMF remained abundant. Seedling biomass production was not correlated with in planta fungal colonization or pathogen load. Soil microbiota can negatively influence baldcypress seedling growth, and no growth benefit was evidenced from the root colonization of mutualist fungi. Flooding can modify baldcypress-fungal interactions by diminishing colonization of DSE. Overall, baldycpress seedlings were more sensitive to the presence of microbiota than flooding, and thus restoration efforts should focus on having a better understanding of plant–microbe interactions in swamps.

Louisiana

Wildfire-resistant biological soil crusts and fire-induced loss of soil stability in Palouse prairies, USA

Frequent low-intensity fires are a natural component of the ecology of the Palouse prairies of northwestern North America. To study the effects of fire upon biological soil crusts (BSCs) occurring in these grasslands, we sampled three burned (in 2000) sites and three unburned sites in the Hell's Canyon area (OR, USA) ???1 year post-fire. We measured vascular plant and BSC cover, soil microbe pigmentation, texture and chemistry, and soil surface physical properties (stability and rugosity). Festuca idahoensis was two times more abundant in unburned plots (P=0.0006), and vascular plant and litter cover were generally higher in unburned plots. At the community scale, there was no difference in the lichen and moss species composition, suggesting much less drastic effects of fire on BSCs than reported in other systems. Soil surface stability (measured using slake value) was significantly lower in burned sites than unburned sites (median value=5 versus 6, P=0.008), a result which is likely due to the greater density of lichens and mosses encountered in the unburned plots. Soil microbe pigmentation was lower in burned plots (P=0.03), suggesting that the biomass of photosynthetic microbes had decreased; however, the presence of intra- and extracellular pigments in burned soils indicates that microorganisms were not eradicated. Pigments most strongly associated with cyanobacteria were more abundant in unburned sites, suggesting that cyanobacteria may have been more strongly impacted by the fire than other BSC components. Composition of nutrients and surface rugosity did not differ significantly between treatments. We hypothesize that Palouse prairie soil crusts are relatively resistant to wildfire because of low fire intensity and their occupation of space away from the vascular plant fuel load.

Applied Soil Ecology

Spatial variation in edaphic characteristics is a stronger control than nitrogen inputs in regulating soil microbial effects on a desert grass

Increased atmospheric nitrogen (N) deposition can have wide-ranging effects on plant community structure and ecosystem function, some of which may be indirectly mediated by soil microbial responses to an altered biogeochemical environment. In this study, soils from a field N fertilization experiment that spanned a soil texture gradient were used as inocula in the greenhouse to assess the indirect effects of soil microbial communities on growth of a desert grass. Plant performance and interaction with soil microbiota were evaluated via plant above- and belowground biomass, leaf N concentration, and root fungal colonization. Nitrogen fertilization in the field increased the benefits of soil microbial inoculation to plant leaf N concentration, but did not alter the effect of soil microbes on plant growth. Plant-microbe interaction outcomes differed most strongly among sites with different soil textures, where the soil microbial community from the sandiest site was most beneficial to host plant growth. The findings of this study suggest that in a desert grassland, increases in atmospheric N deposition may exert a more subtle influence on plant-microbe interactions by altering plant nutrient status, whereas edaphic factors can alter the whole-plant growth response to soil microbial associates.

Journal of Arid Environments

Substantial contribution of biomethylation to aquifer arsenic cycling

Microbes play a prominent role in transforming arsenic to and from immobile forms in aquifers1. Much of this cycling involves inorganic forms of arsenic2, but microbes can also generate organic forms through methylation3, although this process is often considered insignificant in aquifers4, 5, 6, 7. Here we identify the presence of dimethylarsinate and other methylated arsenic species in an aquifer hosted in volcaniclastic sedimentary rocks. We find that dimethylarsinate is widespread in the aquifer and its concentration correlates strongly with arsenite concentration. We use laboratory incubation experiments and an aquifer injection test to show that aquifer microbes can produce dimethylarsinate at rates of about 0.1% of total dissolved arsenic per day, comparable to rates of dimethylarsinate production in surface environments. Based on these results, we estimate that globally, biomethylation in aquifers has the potential to transform 100 tons of inorganic arsenic to methylated arsenic species per year, compared with the 420–1,250 tons of inorganic arsenic that undergoes biomethylation in soils8. We therefore conclude that biomethylation could contribute significantly to aquifer arsenic cycling. Because biomethylation yields arsine and methylarsines, which are more volatile and prone to diffusion than other arsenic species, we further suggest that biomethylation may serve as a link between surface and subsurface arsenic cycling.

Nature Geoscience

Innate immunity is not related to the sex of adult Tree Swallows during the nestling period

Evolutionary theory predicts that exposure to more diverse pathogens will result in the evolution of a more robust immune response. We predicted that during the breeding season the innate immune function of female Tree Swallows ( Tachycineta bicolor ) should be more effective than that of males because (1) the transmission of sexually transmitted microbes during copulation puts females at greater risk because ejaculates move from males to females, (2) females copulate with multiple males, exposing them to the potentially pathogenic microbes in semen, and (3) females spend more time in the nest than do males so may be more exposed to nest microbes and ectoparasites that can be vectors of bacterial and viral pathogens. In addition, elevated testosterone in males may suppress immune function. We tested our prediction during the 2009 breeding season with microbicidal assays in vitro to assess the ability of the innate immune system to kill Escherichia coli . The sexes did not differ in the ability of their whole blood to kill E. coli . We also found no significant relationships between the ability of whole blood to kill E. coli and the reproductive performance or the physical condition of males or females. These results indicate that during the nestling period there are no sexual differences in this component of the innate immune system. In addition, they suggest that there is little association between this component of innate immunity and the reproductive performance and physical condition during the nestling period of adult Tree Swallows.

The Condor