Effects of the nematode Litylenchus crenatae subsp. mccannii and beech leaf disease on leaf fungal and bacterial communities on Fagus grandifolia (American beech).

Beech leaf disease (BLD) is a newly emerging disease in North America that affects American beech (Fagus grandifolia). It is increasingly recognized that BLD is caused by a subspecies of the anguinid nematode Litylenchus crenatae subsp. mccannii (hereafter L. crenatae), which is likely native to East Asia. How nematode infestation of leaves affects the leaf microbiome and whether changes in the microbiome could contribute to BLD symptoms remain uncertain. In this study, we examined bacterial and fungal communities associated with the leaves of F. grandifolia across nine sites in Ohio and Pennsylvania that were either symptomatic or asymptomatic for BLD and used qPCR to measure relative nematode infestation levels. We found significantly higher levels of infestation at sites visibly symptomatic for BLD. Low levels of nematode infestation were also observed at asymptomatic sites, which suggests that nematodes can be present without visible symptoms evident. Bacterial and fungal communities were significantly affected by sampling site and symptomology, but only fungal communities were affected by nematode presence alone. We found many significant indicators of both bacteria and fungi related to symptoms of BLD, with taxa generally occurring in both asymptomatic and symptomatic leaves, suggesting that microbes are not responsible for BLD but could act as opportunistic pathogens. Of particular interest was the fungal genus Erysiphe, which is common in the Fagaceae and is reported to overwinter in buds-a strategy consistent with L. crenatae. The specific role microbes play in opportunistic infection of leaves affected by L. crenatae will require additional study. IMPORTANCE: Beech leaf disease (BLD) is an emerging threat to American beech (Fagus grandifolia) and has spread quickly throughout the northeastern United States and into southern Canada. This disease leads to disfigurement of leaves and is marked by characteristic dark, interveinal banding, followed by leaf curling and drop in more advanced stages. BLD tends to especially affect understory leaves, which can lead to substantial thinning of the forest understory where F. grandifolia is a dominant tree species. Understanding the cause of BLD is necessary to employ management strategies that protect F. grandifolia and the forests where it is a foundation tree species. Current research has confirmed that the foliar nematode Litylenchus crenatae subsp. mccannii is required for BLD, but whether other organisms are involved is currently unknown. Here, we present a study that investigated leaf-associated fungi and bacteria of F. grandifolia to understand more about how microorganisms may contribute to BLD.

Effects of amphibian genetic diversity on ecological communities.

The amount of genetic diversity within a population can affect ecological processes at population, community, and ecosystem levels. However, the magnitude, consistency, and scope of these effects are largely unknown. To investigate these issues, we conducted two experiments manipulating the amount of genetic diversity and environmental factors in larval amphibians. The first experiment manipulated wood frog genetic diversity, the presence or absence of caged predators, and competition from leopard frogs to test whether these factors affected survival, growth, and morphology of wood frogs and leopard frogs. The second experiment manipulated wood frog genetic diversity, the presence or absence of uncaged predators, and resource abundance to test whether these factors affected wood frog traits (survival, morphology, growth, development, and behavior) and other components of the ecological community (zooplankton abundance, phytoplankton, periphyton, and bacterial community structure). Genetic diversity did not affect wood frog survival, growth, and development in either experiment. However, genetic diversity did affect the mean morphology of wood frog tadpoles in the first experiment and the abundance and distribution of zooplankton in the second experiment. It did not affect phytoplankton abundance, periphyton abundance, or bacterial community structure. While effect sizes (Cohen's d) of genetic diversity were approximately half those of environment treatments, the greatest effect sizes were for interaction effects between genetic diversity and environment. Our results indicate that genetic diversity can have a large effect on ecological processes, but the direction of those effects is highly dependent upon environmental conditions, and not easily predicted from simple measures of traits.

Development of Primers Specific for Detection of Litylenchus crenatae, the Causal Agent of Beech Leaf Disease, in Plant Tissue.

Beech leaf disease (BLD), an emerging threat to American beech (Fagus grandifolia) in the northern United States and Canada, was recently confirmed to be caused by the nematode Litylenchus crenatae subsp. mccannii (hereafter L. crenatae). Consequently, there is a need for a rapid, sensitive, and accurate method for detecting L. crenatae for both diagnostic as well as control purposes. This research developed a new set of DNA primers that specifically amplify L. crenatae and allow for accurate detection of the nematode in plant tissue. These primers have also been used in quantitative PCR (qPCR) to determine relative differences in gene copy number between samples. This primer set provides an improved, effective tool for monitoring and detecting L. crenatae in temperate tree leaf tissue which is necessary to understand the spread of this emerging forest pest and to develop management strategies.

The effects of increased hay-to-grain ratio on behavior, metabolic health measures, and fecal bacterial communities in four Masai giraffe (Giraffa camelopardalis tippelskirchi) at Cleveland Metroparks Zoo.

We evaluated whether increasing the hay-to-grain ratio offered to Masai giraffe (Giraffa camelopardalis tippelskirchi) at Cleveland Metroparks Zoo would reduce oral stereotypies and alter feeding behaviors, maintain or increase serum calcium-to-phosphorus ratio, decrease serum insulin-to-glucose ratio and salivary insulin, and alter fecal bacterial community structure. Giraffe transitioned to a ∼90:10 hay-to-grain ratio in even increments over 8 weeks. A ration balancer was added during the seventh week of transition to ensure proper mineral and nutrient balance. We collected (1) behavioral data collected approximately daily using instantaneous focal sampling; (2) serum collections every other week for insulin-to-glucose and calcium-to-phosphorus ratio analysis and saliva weekly for insulin analysis; and (3) weekly fecal sample collections to examine changes in bacterial community structure during the 8 weeks preceding and following the diet change. After the diet change, giraffe spent significantly more time feeding and less time performing tongue and mouth stereotypies, people-directed and alert behaviors. Salivary and serum insulin and serum insulin-to-glucose ratio decreased, and fecal bacterial community structure changed significantly. However, serum calcium-to-phosphorus ratio remained >1:1 throughout the study. While further studies are needed to elucidate the nature and implications of the change in fecal bacterial community structure and metabolic hormones, the results of this study show promise for incremental improvements in health and welfare from feeding a higher proportion of forage in the diet.

Spatial heterogeneity of plant-soil feedbacks increases per capita reproductive biomass of species at an establishment disadvantage.

Plant-soil feedbacks have been widely implicated as a driver of plant community diversity, and the coexistence prediction generated by a negative plant-soil feedback can be tested using the mutual invasibility criterion: if two populations are able to invade one another, this result is consistent with stable coexistence. We previously showed that two co-occurring Rumex species exhibit negative pairwise plant-soil feedbacks, predicting that plant-soil feedbacks could lead to their coexistence. However, whether plants are able to reproduce when at an establishment disadvantage ("invasibility"), or what drivers in the soil might correlate with this pattern, are unknown. To address these questions, we created experimental plots with heterogeneous and homogeneous soils using field-collected conditioned soils from each of these Rumex species. We then allowed resident plants of each species to establish and added invader seeds of the congener to evaluate invasibility. Rumex congeners were mutually invasible, in that both species were able to establish and reproduce in the other's resident population. Invaders of both species had twice as much reproduction in heterogeneous compared to homogeneous soils; thus the spatial arrangement of plant-soil feedbacks may influence coexistence. Soil mixing had a non-additive effect on the soil bacterial and fungal communities, soil moisture, and phosphorous availability, suggesting that disturbance could dramatically alter soil legacy effects. Because the spatial arrangement of soil patches has coexistence implications, plant-soil feedback studies should move beyond studies of mean effects of single patch types, to consider how the spatial arrangement of patches in the field influences plant communities.

Litter chemistry and chemical diversity drive ecosystem processes in forest ponds.

Research suggests that a positive relationship exists between diversity and ecological function, yet the multi-trophic effects of biodiversity remain poorly understood. The resource complementarity hypothesis suggests that increasing the trait diversity of resources provides a more complete diet for consumers, elevating consumer feeding rates. Whereas previous tests of this mechanism have measured trait diversity as the variation of single traits or the richness of functional groups, we employed a multivariate trait index to manipulate the chemical diversity of temperate tree litter species in outdoor pond mesocosms. We inoculated outdoor mesocosms with diverse and multi-trophic communities of microbial and macro-consumer species that rely on leaf litter for energy and nutrients. Litter was provided at three levels of chemical trait diversity, a constant level of species richness, and an equal representation of all litter species. Over three months, we measured more than 65 responses, and assessed the effects of litter chemical diversity and chemical trait means (i.e., community-weighted means). We found that litter chemical diversity positively correlated with decomposition rate of leaf litter, but had no effect on biomass or density of producers and consumers. However, the pond communities often responded to chemical trait means, particularly those related to nutrients, structure, and defense. Our results suggest that resource complementarity does have some effect on the release of energy and nutrients from decomposing substrates in forest ponds, but does not have multi-trophic effects. Our results further suggest that loss of tree biodiversity could affect forest ecosystem functionality, and particularly the processes occurring in and around ponds and wetlands.

Seasonal variation in mycorrhizal fungi colonizing roots of Allium tricoccum (wild leek) in a mature mixed hardwood forest.

The community of arbuscular mycorrhizal (AM) fungi colonizing roots of the forest herb Allium tricoccum Ait. (wild leek) was examined to assess whether colonization varied seasonally and spatially within the forest. Whole plants were collected to coincide with observed phenological stages, and the perennial tissue (i.e., the bulb) was used to analyze total C, N, and P over the growing season. AM fungal community composition, structure, and abundance were assessed in roots by terminal restriction fragment length polymorphism analysis and quantitative PCR. It was found that A. tricoccum rDNA co-amplified using the general AM primers NS31/AM1, and a new primer for qPCR was designed that discriminated against plant DNA to quantify AM colonization. Community structure of AM fungi did not vary seasonally, but did change spatially within the forest, and AM fungal communities were correlated with the presence of overstory tree species. Fungal colonization of roots, however, did change seasonally with a maximum observed in late winter and early spring following leaf emergence. Maximum AM fungal colonization was associated with declines in bulb N and P, suggesting that leaf emergence and growth were responsible for both declines in stored nutrients and increases in AM fungal colonization. Plant N and P contents increased between late summer and early spring while C contents remained unchanged. The observed increase in nutrient content during a time when A. tricoccum lacks leaves indicates that the roots or AM fungi are metabolically active and acquire nutrients during this time, despite an absence of photosynthesis and thus a direct supply of C from A. tricoccum.

Iron Oxide and Titanium Dioxide Nanoparticle Effects on Plant Performance and Root Associated Microbes.

In this study, we investigated the effect of positively and negatively charged Fe3O4 and TiO2 nanoparticles (NPs) on the growth of soybean plants (Glycine max.) and their root associated soil microbes. Soybean plants were grown in a greenhouse for six weeks after application of different amounts of NPs, and plant growth and nutrient content were examined. Roots were analyzed for colonization by arbuscular mycorrhizal (AM) fungi and nodule-forming nitrogen fixing bacteria using DNA-based techniques. We found that plant growth was significantly lower with the application of TiO2 as compared to Fe3O4 NPs. The leaf carbon was also marginally significant lower in plants treated with TiO2 NPs; however, leaf phosphorus was reduced in plants treated with Fe3O4. We found no effects of NP type, concentration, or charge on the community structure of either rhizobia or AM fungi colonizing plant roots. However, the charge of the Fe3O4 NPs affected both colonization of the root system by rhizobia as well as leaf phosphorus content. Our results indicate that the type of NP can affect plant growth and nutrient content in an agriculturally important crop species, and that the charge of these particles influences the colonization of the root system by nitrogen-fixing bacteria.

Shotgun metagenomic analysis of metabolic diversity and microbial community structure in experimental vernal pools subjected to nitrate pulse.

BACKGROUND: Human activities have greatly increased nitrogen (N) levels in natural habitats through atmospheric N deposition and nutrient leaching, which can have large effects on N cycling and other ecosystem processes. Because of the significant role microorganisms play in N cycling, high inputs of nitrogenous compounds, such as nitrate (NO3-), into natural ecosystems could have cascading effects on microbial community structure and the metabolic processes that microbes perform. To investigate the multiple effects of NO3- pollution on microbial communities, we created two shotgun metagenomes from vernal pool microcosms that were either enriched with a solution of 10 mg NO3--N (+NO3-) or received distilled water as a control (-N). RESULTS: After only 20 hours of exposure to NO3-, the initial microbial community had shifted toward one containing a higher proportional abundance of stress tolerance and fermentation environmental gene tags (EGTs). Surprisingly, we found no changes to N metabolism EGTs, even though large shifts in denitrification rates were seen between the +NO3- and -N microcosms. Thus, in the absence of NO3- addition, it is plausible that the microbes used other respiratory pathways for energy. Respiratory pathways involving iron may have been particularly important in our -N microcosms, since iron acquisition EGTs were proportionally higher in the -N metagenome. Additionally, we noted a proportional increase in Acidobacteria and Alphaproteobacteria EGTs in response to NO3- addition. These community shifts in were not evident with TRFLP, suggesting that metagenomic analyses may detect fine-scale changes not possible with community profiling techniques. CONCLUSIONS: Our results suggest that the vernal pool microbial communities profiled here may rely on their metabolic plasticity for growth and survival when certain resources are limiting. The creation of these metagenomes also highlights how little is known about the effects of NO3- pollution on microbial communities, and the relationship between community stability and function in response to disturbance.

Belowground biotic complexity drives aboveground dynamics: a test of the soil community feedback model.

Feedbacks between soil communities and plants may determine abundance and diversity in plant communities by influencing fitness and competitive outcomes. We tested the core hypotheses of soil community feedback theory: plant species culture distinct soil communities that alter plant performance and the outcome of interspecific competition. We applied this framework to inform the repeated dominance of Solidago canadensis in old-field communities. In glasshouse experiments, we examined the effects of soil communities on four plant species' performance in monoculture and outcomes of interspecific competition. We used terminal restriction fragment length polymorphism (TRFLP) analysis to infer differences in the soil communities associated with these plant species. Soil community origin had strong effects on plant performance, changed the intensity of interspecific competition and even reversed whether plant species were limited by conspecifics or heterospecifics. These plant-soil feedbacks are strong enough to upend winners and losers in classic competition models. Plant species cultured significantly different mycorrhizal fungal and bacterial soil communities, indicating that these feedbacks are likely microbiotic in nature. In old-fields and other plant communities, these soil feedbacks appear common, fundamentally alter the intensity and nature of plant competition and potentially maintain diversity while facilitating the dominance of So. canadensis.

Interannual variation in spring weather conditions as a driver of spring wildflower coverage: a 15-year perspective from an old-growth temperate forest.

Spring ephemerals are wildflowers found in temperate deciduous forests that typically display aboveground shoots for a period of 2 months or less. Early spring, before the canopy leaves out, marks the beginning of the aboveground growth period where ephemerals acquire nutrients and resources via aboveground tissues. Several studies have shown that spring ephemeral reproduction is affected by spring temperature, but few have looked at how weather conditions of the current and previous seasons, including precipitation and temperature, influence aboveground growth. Here, we examine the response of a spring ephemeral community in a temperate hardwood forest to weather conditions during their current and previous growing seasons. For 15 years we estimated percent cover of each species within our community. We highlighted five dominant spring ephemerals within this community: wild leek (Allium tricoccum), cutleaf toothwort (Cardamine concatenata), spring beauty (Claytonia virginica), squirrel corn (Dicentra canadensis) and trout lily (Erythronium americanum). We compared changes in cover on both a community and species level from 1 year to the next with average precipitation and temperature of the year of measurement as well as the year prior. We found precipitation and temperature influence a change in cover at the community and species level, but the strength of that influence varies by species. There were few significant correlations between plant cover in the current year and temperature and precipitation in the 30 days preceding measurement. However, we found significant correlations between plant cover and precipitation and temperature during the previous spring; precipitation and cover change were positively correlated, whereas temperature and cover change were negatively correlated. Overall, cooler, wetter springs lead to an increase in aboveground cover the next year. Learning how individual species within a forest plant community respond to weather conditions is a crucial part of understanding how plant communities will respond to climate change.

Beech Leaf Disease Severity Affects Ectomycorrhizal Colonization and Fungal Taxa Composition.

Beech leaf disease (BLD) is an emerging forest infestation affecting beech trees (Fagus spp.) in the midwestern and northeastern United States and southeastern Canada. BLD is attributed to the newly recognized nematode Litylenchus crenatae subsp. mccannii. First described in Lake County, Ohio, BLD leads to the disfigurement of leaves, canopy loss, and eventual tree mortality. Canopy loss limits photosynthetic capacity, likely impacting tree allocation to belowground carbon storage. Ectomycorrhizal fungi are root symbionts, which rely on the photosynthesis of autotrophs for nutrition and growth. Because BLD limits tree photosynthetic capacity, ECM fungi may receive less carbohydrates when associating with severely affected trees compared with trees without BLD symptoms. We sampled root fragments from cultivated F. grandifolia sourced from two provenances (Michigan and Maine) at two timepoints (fall 2020 and spring 2021) to test whether BLD symptom severity alters colonization by ectomycorrhizal fungi and fungal community composition. The studied trees are part of a long-term beech bark disease resistance plantation at the Holden Arboretum. We sampled from replicates across three levels of BLD symptom severity and compared fungal colonization via visual scoring of ectomycorrhizal root tip abundance. Effects of BLD on fungal communities were determined through high-throughput sequencing. We found that ectomycorrhizal root tip abundance was significantly reduced on the roots of individuals of the poor canopy condition resulting from BLD, but only in the fall 2020 collection. We found significantly more ectomycorrhizal root tips from root fragments collected in fall 2020 than in spring 2021, suggesting a seasonal effect. Community composition of ectomycorrhizal fungi was not impacted by tree condition but did vary between provenances. We found significant species level responses of ectomycorrhizal fungi between levels of both provenance and tree condition. Of the taxa analyzed, two zOTUs had significantly lower abundance in high-symptomatology trees compared with low-symptomatology trees. These results provide the first indication of a belowground effect of BLD on ectomycorrhizal fungi and contribute further evidence to the role of these root symbionts in studies of tree disease and forest pathology.

Bioretention column study of bacteria community response to salt-enriched artificial stormwater.

Cold climate cities with green infrastructure depend on soil bacteria to remove nutrients from road salt-enriched stormwater. Our research examined how bacterial communities in laboratory columns containing bioretention media responded to varying concentrations of salt exposure from artificial stormwater and the effect of bacteria and salt on column effluent concentrations. We used a factorial design with two bacteria treatments (sterile, nonsterile) and three salt concentrations (935, 315, and 80 ppm), including a deionized water control. Columns were repeatedly saturated with stormwater or deionized and then drained throughout 5 wk, with the last week of effluent analyzed for water chemistry. To examine bacterial communities, we extracted DNA from column bioretention media at time 0 and at week 5 and used molecular profiling techniques to examine bacterial community changes. We found that bacterial community taxa changed between time 0 and week 5 and that there was significant separation between taxa among salt treatments. Bacteria evenness was significantly affected by stormwater treatment, but there were no differences in bacterial richness or diversity. Soil bacteria and salt treatments had a significant effect on the effluent concentration of NO, PO, Cu, Pb, and Zn based on ANOVA tests. The presence of bacteria reduced effluent NO and Zn concentrations by as much as 150 and 25%, respectively, while having a mixed effect on effluent PO concentrations. Our results demonstrate how stormwater can affect bacterial communities and how the presence of soil bacteria improves pollutant removal by green infrastructure.

Effects of the invasive plant garlic mustard (Alliaria petiolata) on bacterial communities in a northern hardwood forest soil.

We compared the effects of the invasive plant Alliaria petiolata (garlic mustard) and 2 native plants on soil bacterial communities in a mature mesophytic forest. Soil samples were collected from plant patches containing either Alliaria or the native plants Allium tricoccum (wild leek) and Gallium triflorum (bedstraw). Since Alliaria litter contains secondary compounds that have reported antimicrobial properties, soil was collected outside the root zone of the plants but within the plant patches such that the soil would have been influenced by the litter of the respective plant species but not by plant roots. DNA was extracted from the soil samples and used to amplify the 16S rRNA gene region using bacterial specific primers. Terminal restriction fragment length polymorphism (TRFLP) profiles of each bacterial community were used to examine differences in bacterial communities among the plant species and between August and April sampling. Bacterial richness, evenness, and diversity were not significantly affected by plant species. Non-metric multidimensional scaling (NMS) suggested that differences existed between August and April sampling, but that plant species litter exerted a much weaker effect on soil bacterial communities. Soil physiochemical conditions were significantly correlated with soil bacterial communities and may underlie the observed seasonal changes in bacterial communities.

Vegetation and soil environment influence the spatial distribution of root-associated fungi in a mature beech-maple forest.

Although the level of diversity of root-associated fungi can be quite high, the effect of plant distribution and soil environment on root-associated fungal communities at fine spatial scales has received little attention. Here, we examine how soil environment and plant distribution affect the occurrence, diversity, and community structure of root-associated fungi at local patch scales within a mature forest. We used terminal restriction fragment length polymorphism and sequence analysis to detect 63 fungal species representing 28 different genera colonizing tree root tips. At least 32 species matched previously identified mycorrhizal fungi, with the remaining fungi including both saprotrophic and parasitic species. Root fungal communities were significantly different between June and September, suggesting a rapid temporal change in root fungal communities. Plant distribution affected root fungal communities, with some root fungi positively correlated with tree diameter and herbaceous-plant coverage. Some aspects of the soil environment were correlated with root fungal community structure, with the abundance of some root fungi positively correlated with soil pH and moisture content in June and with soil phosphorous (P) in September. Fungal distribution and community structure may be governed by plant-soil interactions at fine spatial scales within a mature forest. Soil P may play a role in structuring root fungal communities at certain times of the year.

Utilizing herbarium specimens to quantify historical mycorrhizal communities.

PREMISE OF THE STUDY: Mycorrhiza are critical to ecosystem functioning, but a lack of historical baseline data limits our understanding of the long-term belowground effects of global change. Herbarium specimens may provide this needed insight. However, it is unknown whether DNA of arbuscular mycorrhizal fungi (AMF) can be reliably extracted from vascular plant specimen roots. METHODS: We sampled roots from herbarium specimens of four herbaceous forest species collected in western Pennsylvania between 1881-2008. Using molecular methods (terminal restriction fragment length polymorphism and sequence analysis), we quantified AMF communities from specimen roots and tested for contamination. RESULTS: We successfully amplified AMF DNA from 44% (21/48) of the root but not leaf samples, indicating specimen contamination was negligible. As expected, there were significant differences in AMF composition between plant species (P < 0.05). However, no differences in AMF communities were detected through time, possibly due to limited sample size and low amplification rates in recent collections. DISCUSSION: Herbaria have potential as sources of valuable belowground microbial data to answer questions across geographic, temporal, and taxonomic scales otherwise not feasible. Ongoing methodological developments will only magnify this potential. Further tests are needed to determine curatorial practices that maximize this innovative use of herbarium specimens.

Molecular analysis of bacterial communities associated with the roots of Douglas fir (Pseudotsuga menziesii) colonized by different ectomycorrhizal fungi.

We studied the effect of ectomycorrhizal fungi on bacterial communities colonizing roots of Douglas fir (Pseudotsuga menziesii). Mycorrhizal tips were cleaned of soil and separated based on gross morphological characteristics. Sequencing of the internal transcribed spacers of the nuclear rRNA gene cluster indicated that the majority of the tips were colonized by fungi in the Russulaceae, with the genera Russula and Lactarius comprising 70% of the tips. Because coamplification of organellar 16S rRNA genes can interfere with bacterial community analysis of root tips, we developed and tested a new primer pair that permits amplification of bacterial 16S rRNA genes but discriminates more effectively against organellar sequences than commonly used bacterial primer sets. We then used terminal restriction fragment length polymorphism (T-RFLP) and sequence analysis of the 16S rRNA gene to examine differences in bacterial communities associated with the mycorrhizal tips. Cluster analysis of T-RFLP profiles indicated that there were different bacterial communities among the root tips; however, the communities did not seem to be affected by the taxonomic identity of the ectomycorrhizal fungi. Terminal restriction fragment profiling and sequencing of cloned partial 16S rRNA genes indicated that most bacteria on the ectomycorrhizal tips were related to the Alphaproteobacteria and the Bacteroidetes group.

Analysis of nitrogen-fixing members of the epsilon subclass of Proteobacteria in salt marsh sediments.

Based on phylogenetic analysis of clones retrieved from two nifH gene clone libraries that were created using cDNA from suboxic sediment samples obtained from areas densely vegetated with the high-salt marsh plant Spartina patens, a primer set was designed to target nitrogen-fixing bacteria with sequence similarities to members of the epsilon subclass of Proteobacteria. Nested PCR, denaturing gel electrophoresis, and subsequent sequence analysis of reamplified fragments confirmed the specificity of the primer set by retrieving nifH sequences of only putative members of the epsilon subclass of Proteobacteria, all of which were characterized by a highly divergent 27- or 36-bp insertion in both DNA and cDNA.

Fate and effects of heavy metals in salt marsh sediments.

The fate and effects of selected heavy metals were examined in sediment from a restored salt marsh. Sediment cores densely covered with Spartina patens were collected and kept either un-amended or artificially amended with nickel (Ni) under standardized greenhouse conditions. Ni-amendment had no significant effect on the fate of other metals in sediments, however, it increased root uptake of the metals. Metal translocation into the shoots was small for all metals. Higher Ni concentrations in plants from amended cores were accompanied by seasonal reductions in plant biomass, photosynthetic capacity and transfer efficiency of open photosystem II reaction centers; these effects, however, were no longer significant at the end of the growing season. Root colonization by arbuscular mycorrhizal fungi (AMF) resembled that of natural salt marshes with up to 20% root length colonized. Although Ni-amendment increased AMF colonization, especially during vegetative growth, in general AMF were largely unaffected.

Gut microbiome composition is associated with cardiac disease in zoo-housed western lowland gorillas (Gorilla gorilla gorilla).

Cardiac disease is a leading cause of mortality in zoo-housed western lowland gorillas (Gorilla gorilla gorilla). The gut microbiome is associated with cardiac disease in humans and similarly the gut microbiome may be associated with cardiac diseases in close relatives of humans, such as gorillas. We assessed the relationship between cardiac disease and gut bacterial composition in eight zoo-housed male western lowland gorillas (N = 4 with and N = 4 without cardiac disease) utilizing 16S rRNA gene analysis on the Illumina MiSeq sequencing platform. We found bacterial composition differences between gorillas with and without cardiac disease. Bacterial operational taxonomic units from phyla Bacteroidetes, Spirochaetes, Proteobacteria and Firmicutes were significant indicators of cardiac disease. Our results suggest that further investigations between diet and cardiac disease could improve the management and health of zoo-housed populations of this endangered species.