Microbial diversity and diazotrophy associated with the freshwater non-heterocyst forming cyanobacterium Lyngbya robusta

Lake Atitlan, Guatemala, a freshwater lake in South America, experiences annually recurring blooms comprised of the planktic filamentous cyanobacterium Lyngbya robusta. Previous physiochemical characterisation of the bloom identified diurnal nitrogenase activity typical of non-heterocystous cyanobacteria, in addition to the low-level detection of the cyanotoxins cylindrospermopsin and saxitoxin. A molecular approach, combining deep sequencing of the 16S rRNA and nifH genes, was applied to a cyanobacteria-dominated sample collected during the extensive 2009 bloom. Lyngbya accounted for over 60 % of the total 16S rRNA sequences with the only other cyanobacterial species detected being the picophytoplankton Synechococcus. The remaining bacterial population was comprised of organisms typical of other eutrophic freshwater bodies, although the proportionate abundances were atypical. An obligate anaerobe Opitutus, not typically found in freshwater systems, was identified within the community which suggests it may have a role in enhancing nitrogen fixation. Primary nitrogen fixation was attributed to Lyngbya, with other putative nitrogen fixers, Desulfovibrio, Clostridium and Methylomonas, present at very low abundance.     

Deep sequencing of non-ribosomal peptide synthetases and polyketide synthases from the microbiomes of Australian marine sponges

The biosynthesis of non-ribosomal peptide and polyketide natural products is facilitated by multimodular enzymes that contain domains responsible for the sequential condensation of amino and carboxylic subunits. These conserved domains provide molecular targets for the discovery of natural products from microbial metagenomes. This study demonstrates the application of tag-encoded FLX amplicon pyrosequencing (TEFAP) targeting non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) genes as a method for determining the identity and diversity of natural product biosynthesis genes. To validate this approach, we assessed the diversity of NRPS and PKS genes within the microbiomes of six Australian marine sponge species using both TEFAP and metagenomic whole-genome shotgun sequencing approaches. The TEFAP approach identified 100 novel ketosynthase (KS) domain sequences and 400 novel condensation domain sequences within the microbiomes of the six sponges. The diversity of KS domains within the microbiome of a single sponge species Scopalina sp. exceeded that of any previously surveyed marine sponge. Furthermore, this study represented the first to target the condensation domain from NRPS biosynthesis and resulted in the identification of a novel condensation domain lineage. This study highlights the untapped potential of Australian marine sponges for the isolation of novel bioactive natural products. Furthermore, this study demonstrates that TEFAP approaches can be applied to functional genes, involved in natural product biosynthesis, as a tool to aid natural product discovery. It is envisaged that this approach will be used across multiple environments, offering an insight into the biological processes that influence the production of secondary metabolites.     

Discordant risk: Overweight and cardiometabolic risk in Chinese adults

Objective:

Recent US work identified “metabolically healthy overweight” and “metabolically at risk normal weight” individuals. Less is known for modernizing countries with recent increased obesity.

Design and Methods:

Fasting blood samples, anthropometry and blood pressure from 8,233 adults aged 18‐98 in the 2009 nationwide China Health and Nutrition Survey, were used to determine prevalence of overweight (Asian cut point, BMI ≥23 kg/m²) and five risk factors (prediabetes/diabetes (hemoglobin A1c ≥5.7%) inflammation (high‐sensitivity C‐reactive protein (hsCRP) ≥3 mg/l), prehypertension/hypertension (Systolic blood pressure/diastolic blood pressure≥130/85 mm Hg), high triglycerides (≥150 mg/dl), low high‐density lipoprotein cholesterol (<40 (men)/ <50 mg/dl (women)). Sex‐stratified, logistic, and multinomial logistic regression models estimated concurrent obesity and cardiometabolic risk, with and without abdominal obesity, adjusting for age, smoking, alcohol consumption, physical activity, urbanicity, and income.

Results:

Irrespective of urbanicity, 78.3% of the sample had ≥1 elevated cardiometabolic risk factor (normal weight: 33.2% had ≥1 elevated risk factor; overweight: 5.7% had none). At the age of 18‐30 years, 47.4% had no elevated risk factors, which dropped to 6% by the age 70, largely due to age‐related increase in hypertension risk (18‐30 years: 11%; >70 years: 73%). Abdominal obesity was highly predictive of metabolic risk, irrespective of overweight (e.g., “metabolically at risk overweight” relative to “metabolically healthy normal weight” (men: relative risk ratio (RRR) = 39.06; 95% confidence interval (CI): 23.47, 65.00; women: RRR = 22.26; 95% CI: 17.49, 28.33)).

Conclusion:

A large proportion of Chinese adults have metabolic abnormalities. High hypertension risk with age, underlies the low prevalence of metabolically healthy overweight. Screening for cardiometabolic‐related outcomes dependent upon overweight will likely miss a large portion of the Chinese at risk population.     

Pressuring and restrictive feeding styles influence infant feeding and size among a low‐income African‐American sample

Objective:

The prevalence of overweight among infants and toddlers has increased dramatically in the past three decades, highlighting the importance of identifying factors contributing to early excess weight gain, particularly in high‐risk groups. Parental feeding styles and the attitudes and behaviors that characterize parental approaches to maintaining or modifying children's eating behavior are an important behavioral component shaping early obesity risk.

Design and Methods:

Using longitudinal data from the Infant Care and Risk of Obesity Study, a cohort study of 217 African‐American mother‐infant pairs with feeding styles, dietary recalls, and anthropometry collected from 3 to 18 months of infant age, we examined the relationship between feeding styles, infant diet, and weight‐for‐age and sum of skinfolds.

Results:

Longitudinal mixed models indicated that higher pressuring and indulgent feeding style scores were positively associated with greater infant energy intake, reduced odds of breastfeeding, and higher levels of age‐inappropriate feeding of liquids and solids, whereas restrictive feeding styles were associated with lower energy intake, higher odds of breastfeeding, and reduced odds of inappropriate feeding. Pressuring and restriction were also oppositely related to infant size with pressuring associated with lower infant weight‐for‐age and restriction with higher weight‐for‐age and sum of skinfolds. Infant size also predicted maternal feeding styles in subsequent visits indicating that the relationship between size and feeding styles is likely bidirectional.

Conclusion:

Our results suggest that the degree to which parents are pressuring or restrictive during feeding shapes the early feeding environment and, consequently, may be an important environmental factor in the development of obesity.     

Quantification of mesocosm fish and amphibian species diversity via environmental DNA metabarcoding

Freshwater fauna are particularly sensitive to environmental change and disturbance. Management agencies frequently use fish and amphibian biodiversity as indicators of ecosystem health and a way to prioritize and assess management strategies. Traditional aquatic bioassessment that relies on capture of organisms via nets, traps and electrofishing gear typically has low detection probabilities for rare species and can injure individuals of protected species. Our objective was to determine whether environmental DNA (eDNA) sampling and metabarcoding analysis can be used to accurately measure species diversity in aquatic assemblages with differing structures. We manipulated the density and relative abundance of eight fish and one amphibian species in replicated 206‐L mesocosms. Environmental DNA was filtered from water samples, and six mitochondrial gene fragments were Illumina‐sequenced to measure species diversity in each mesocosm. Metabarcoding detected all nine species in all treatment replicates. Additionally, we found a modest, but positive relationship between species abundance and sequencing read abundance. Our results illustrate the potential for eDNA sampling and metabarcoding approaches to improve quantification of aquatic species diversity in natural environments and point the way towards using eDNA metabarcoding as an index of macrofaunal species abundance.     

Unravelling core microbial metabolisms in the hypersaline microbial mats of Shark Bay using high-throughput metagenomics

Modern microbial mats are potential analogues of some of Earth''s earliest ecosystems. Excellent examples can be found in Shark Bay, Australia, with mats of various morphologies. To further our understanding of the functional genetic potential of these complex microbial ecosystems, we conducted for the first time shotgun metagenomic analyses. We assembled metagenomic next-generation sequencing data to classify the taxonomic and metabolic potential across diverse morphologies of marine mats in Shark Bay. The microbial community across taxonomic classifications using protein-coding and small subunit rRNA genes directly extracted from the metagenomes suggests that three phyla Proteobacteria, Cyanobacteria and Bacteriodetes dominate all marine mats. However, the microbial community structure between Shark Bay and Highbourne Cay (Bahamas) marine systems appears to be distinct from each other. The metabolic potential (based on SEED subsystem classifications) of the Shark Bay and Highbourne Cay microbial communities were also distinct. Shark Bay metagenomes have a metabolic pathway profile consisting of both heterotrophic and photosynthetic pathways, whereas Highbourne Cay appears to be dominated almost exclusively by photosynthetic pathways. Alternative non-rubisco-based carbon metabolism including reductive TCA cycle and 3-hydroxypropionate/4-hydroxybutyrate pathways is highly represented in Shark Bay metagenomes while not represented in Highbourne Cay microbial mats or any other mat forming ecosystems investigated to date. Potentially novel aspects of nitrogen cycling were also observed, as well as putative heavy metal cycling (arsenic, mercury, copper and cadmium). Finally, archaea are highly represented in Shark Bay and may have critical roles in overall ecosystem function in these modern microbial mats.     

In Situ Measurement of Solid Electrolyte Interphase Evolution on Silicon Anodes Using Atomic Force Microscopy

In situ measurements of the growth of solid electrolyte interphase (SEI) layer on silicon and the lithiation‐induced volume changes in silicon in lithium ion half‐cells are reported. Thin film amorphous silicon electrodes are fabricated in a configuration that allows unambiguous separation of the total thickness change into contribution from SEI thickness and silicon volume change. Electrodes are assembled into a custom‐designed electrochemical cell, which is integrated with an atomic force microscope. The electrodes are subjected to constant potential lithiation/delithiation at a sequence of potential values and the thickness measurements are made at each potential after equilibrium is reached. Experiments are carried out with two electrolytes—1.2 m lithium hexafluoro‐phosphate (LiPF₆) in ethylene carbonate (EC) and 1.2 m LiPF₆ in propylene carbonate (PC)—to investigate the influence of electrolyte composition on SEI evolution. It is observed that SEI formation occurs predominantly during the first lithiation and the maximum SEI thickness is ≈17 and 10 nm respectively for EC and PC electrolytes. This study also presents the measured Si expansion ratio versus equilibrium potential and charge capacity versus equilibrium potential; both relationships display hysteresis, which is explained in terms of the stress–potential coupling in silicon.