bioRxiv Science⌕ Search

Biology subjects

Ottesen, E. A.

Publications and source records attributed to Ottesen, E. A..

8 recordsLinked to original sources

Active non-redundancy and viral orchestration sustain diel microbial successions in the coastal ocean

Ecosystem resilience in dynamic coastal oceans is conventionally ascribed to functional redundancy, where taxonomically distinct microbes buffer environmental fluctuations through interchangeable metabolic roles. In this study, we reveal a deterministic succession architecture sustained by active non redundancy and temporal metabolic coupling, uncovered via autonomous drone array metatranscriptomic sampling in Daya Bay, China. By resolving the transcriptional landscape into six chronometrically phased modules rather than simple time points, we demonstrate a near total renewal of the active gene pool, with greater than 90% of transcribed clusters showing phase exclusivity within approximatively a two-hour window. This radical functional reshaping exposes a tight, scale dependent taxonomic functional coupling where community composition is a strong, linear predictor of metabolic output, peaking at the genus level (p < 0.0001). Functional continuity is maintained not by redundant generalists, but by a precisely sequential relay of specialists, each optimized for transient microniches with minimal overlap between successive phases. Deep learning structural adjudication of the psbA gene pool further reveals viral orchestration of this temporal coupling, where incoming cyanophages introduce structurally distinct protein variants that sustain photosynthetic electron flow under peak irradiance. These findings redefine coastal microbiome stability as a fine-tuned rapid succession of temporal specialists rather than a redundant backdrop. We propose a fundamental revision of marine ecosystem models, shifting from passive buffering frameworks to deterministic, clock driven architectures, which may prove to be critical for forecasting microbiome responses to accelerating short term climate variability.

microbiology↗

Niche specialization and cross-feeding interactions shaping gut microbial fiber degradation in a model omnivore

1The gut microbiome plays an active role in host health, producing gut metabolites that influence host digestive and immune function while also mediating microbial crosstalk. Dietary fiber is a major source of important fermentation byproducts that are generally implicated in gut community stability and host wellbeing, but dissecting microbe-specific contributions to polysaccharide metabolism in the context of a complex gut community is challenging with conventional model organisms. Using the American cockroach (Periplaneta americana) as a model omnivore, we use chemically-defined synthetic diets to identify how complex gut microbial communities respond to two of the most abundant plant polysaccharides, xylan and cellulose. To do so, we fed cockroaches synthetic diets containing one of these fibers or a mix of both in differing ratios. Through both 16S rRNA gene profiling and RNA-seq, we show that mixed fibers enrich for organisms characteristic of the source fibers as well as additional organisms only enriched in mixed-fiber diets. Through an organism-centric pangenome approach, we identify the impact of these fibers on gut microbiome activity. We found that gut communities responded strongly to xylan, with Bacteroidota belonging to Bacteroides, Dysgonomonas, and Parabacteroides producing xylan-active CAZymes at high levels. Multiple groups of Bacillota also responded strongly to a xylan diet, but appeared to act as cross-feeding secondary degraders, producing primarily xylosidases and transcripts associated with xylose utilization. In contrast, cellulose diets were associated with higher transcriptional activity among Fibrobacterota, which are typically a minor component of the cockroach gut microbiome but were the primary producers of CAZymes associated with cellulose and cellobiose degradation. These experiments provide new insight into gut microbial metabolism of these complex plant polysaccharides. Further, they highlight the utility of the cockroach model and synthetic diets to answer fundamental questions about gut microbial responses to different polysaccharides alone and in combination.

microbiology↗

Large temperature excursions have modest impacts on community composition in the high diversity gut microbiome of omnivorous American cockroaches (Periplaneta americana)

Microbial residents of ectothermic hosts are exposed to variations in temperature that have the potential to impact their physiology and the host-microbe symbiotic relationship. In this experimental warming study, laboratory populations of American cockroaches (Periplaneta americana) were kept at a baseline low room temperature of 20-22{degrees}C or a high temperature of 30{degrees}C for two weeks. We quantified bacterial load and performed high-throughput 16S rRNA gene sequencing to assess the hindgut microbiomes response to a near 10{degrees}C shift in environmental temperature. We report modest impacts of temperature on cockroach gut microbiome composition. The high temperature treatment induced increases in the relative abundance of Proteobacteria and Euryarchaeota phyla as well as the Lactobacillaceae and Enterococcaceae families. We also observed increased interindividual variability. There were no significant differences in the dominant Bacteroidota or Firmicutes phyla and no significant losses or reductions in taxa or bacterial load, respectively. This suggests that the gut community of American cockroaches is largely resilient to prolonged increases in temperature and has implications for the cockroach to withstand the impacts of climate change. ImportanceInsects, as with most animals, often harbor microbial symbionts that play an essential role in host health and nutrition. As insects are ectotherms, these microbial symbionts are subject to the same temperature fluctuations as their hosts, potentially impacting host temperature responses. Here, we demonstrate that the American cockroach (Periplaneta americana) gut microbiome exhibits only modest changes following an [~]10{degrees}C increase in environmental temperature. This contrasts with studies in other insects, whose microbiota were highly responsive to temperature variation. This work illustrates that the microbiota of insects may vary in their sensitivity to long-term temperature shifts, providing a more comprehensive understanding of potential variability in insect responses to climate change.

microbiology↗

Genome Report: Improved chromosome-level genome assembly of the American cockroach, Periplaneta americana

1The American cockroach, Periplaneta americana, is a cosmopolitan insect notorious for thriving among humans undeterred by attempts to eliminate it. The traits that contribute to its ubiquity as an opportunistic pest, such as long lifespan, expansive neurosensory capacity, and nutritional flexibility, also make P. americana an excellent invertebrate model organism with a long history in neuroscience and physiological research. Current genetic resources available for P. americana highlight its large, complex genome and richly diverse transcriptional capabilities, but fall short of producing a complete, chromosome-level genome. Here, we present a high-quality de novo genome assembly of a laboratory-raised adult female P. americana using a combination of high fidelity PacBio long reads and Hi-C sequencing. The final 3.23 Gb genome was assembled with chromosomal resolution into 17 scaffolds, consistent with previous karyotype analysis, and has a scaffold N50 of 188.1 Mb and genome BUSCO score of 99.7%. This assembly includes a chromosome that was missing from the previous reference genome for this species. Protein prediction and annotation were performed via the NCBI Eukaryotic Genome Annotation Pipeline, which identified 16,780 protein-coding genes and generated an annotation BUSCO score of 97.8%. Ortholog comparisons with available Blattodea assemblies highlight the expanded chemosensory and immune capabilities of P. americana compared to termite relatives. This genome assembly is a valuable tool for facilitating future research on the biology and evolution of this remarkable insect.

genomics↗

Microbial transcriptional responses to host diet maintain gut microbiome homeostasis in the American cockroach

Scavenging omnivores face challenges in maintaining gut microbiome homeostasis due to a highly variable diet. Work in mammalian models has highlighted the importance of fiber-degrading taxa and their utilization of dietary polysaccharides and host glycans in structuring gut microbiome composition and function. However, less is known about biological mechanisms supporting gut microbiome homeostasis in insects. Here, we present a metatranscriptomic analysis of hindgut microbial transcriptional responses to diverse host diets in the American cockroach (Periplaneta americana), a hindgut-fermenting and omnivorous insect. We analyzed microbial community transcriptomes from hindguts of cockroaches fed high-fiber, high-protein, high-fat, and starvation diets. Microbial taxa predicted to be primary fiber degraders increased transcription of genes associated with utilization of dietary polysaccharides in insects fed a high-fiber diet. In contrast, these taxa responded to starvation and low fiber diets with upregulation of transcripts associated with degradation of host glycans. Analysis of N- and O-glycans in the cockroach midgut confirm the presence of predicted substrates for these pathways. As a result, we hypothesize that peritrophic matrix glycans serve as a critical carbohydrate subsidy supporting insect gut microbial metabolism in the absence of dietary fiber. Observed stability in gut microbiome composition and the transcriptional activity of many secondary fermenters and hydrogenotrophic taxa in the gut suggests that this strategy can be highly successful. Together, this work supports a novel physiological role for the peritrophic matrix of insects and other invertebrates in supporting gut microbiome homeostasis across dietary shifts.

microbiology↗

Synthetic diets containing a single polysaccharide disrupt gut microbial community structure and microbial interaction networks in the American cockroach.

1The macronutrient composition of a hosts diet shapes its gut microbial community, with dietary fiber in particular escaping host digestion to serve as a potent carbon source for gut microbiota. Despite widespread recognition of fibers importance to microbiome health, nutritional research often fails to differentiate hyper-processed fibers from cell-matrix derived intrinsic fibers, limiting our understanding of how individual polysaccharides influence the gut community. We use the American cockroach (Periplaneta americana) as a model system to dissect the response of complex gut microbial communities to diet modifications that are impossible to test in traditional host models. Here, we designed synthetic diets from lab-grade, purified ingredients to identify how the cockroach microbiome responds to six different carbohydrates (chitin, methylcellulose, microcrystalline cellulose, pectin, starch, xylan) in otherwise balanced diets. We show via 16S rRNA gene profiling that these synthetic diets reduce bacterial diversity and alter the phylogenetic composition of cockroach gut microbiota in a fiber-dependent manner, regardless of the vitamin and protein content of the diet. Comparisons with cockroaches fed whole-food diets reveal that synthetic diets induce blooms in common cockroach-associated taxa and subsequently fragment previously stable microbial correlation networks. Our research leverages an unconventional microbiome model system and customizable lab-grade artificial diets to shed light on how purified polysaccharides, as opposed to nutritionally complex intrinsic fibers, exert substantial influence over a normally stable gut community.

microbiology↗

Mild and severe SARS-CoV-2 infection induces respiratory and intestinal microbiome changes in the K18-hACE2 transgenic mouse model

Transmission of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in millions of deaths and declining economies around the world. K18-hACE2 mice develop disease resembling severe SARS-CoV-2 infection in a virus dose-dependent manner. The relationship between SARS-CoV-2 and the intestinal or respiratory microbiome is not fully understood. In this context, we characterized the cecal and lung microbiome of SARS-CoV-2 challenged K18-hACE2 transgenic mice in the presence or absence of treatment with the Mpro inhibitor GC376. Cecum microbiome showed decreased Shannon and Inv Simpson diversity index correlating with SARS-CoV-2 infection dosage and a difference of Bray-Curtis dissimilarity distances among control and infected mice. Bacterial phyla such as Firmicutes, particularly Lachnospiraceae and Oscillospiraceae, were significantly less abundant while Verrucomicrobiota, particularly the family Akkermansiaceae, were increasingly more prevalent during peak infection in mice challenged with a high virus dose. In contrast to the cecal microbiome, the lung microbiome showed similar microbial diversity among the control, low and high challenge virus groups, independent of antiviral treatment. Bacterial phyla in the lungs such as Bacteroidota decreased while Firmicutes and Proteobacteria were significantly enriched in mice challenged with a high dose of SARS-CoV-2. In summary, we identified changes in the cecal and lung microbiome of K18-hACE2 mice with severe clinical signs of SARS-CoV-2 infection. IMPORTANCEThe COVID-19 pandemic has resulted in millions of deaths. The hosts respiratory and intestinal microbiome can affect directly or indirectly the immune system during viral infections. We characterized the cecal and lung microbiome in a relevant mouse model challenged with a low and high dose of SARS-CoV-2 in the presence or absence of an antiviral Mpro inhibitor, GC376. Decreased microbial diversity and taxonomic abundances of the phyla Firmicutes, particularly Lachnospiraceae, correlating with infection dosage was observed in the cecum. In addition, microbes within the family Akkermansiaceae were increasingly more prevalent during peak infection, which is observed in other viral infections. The lung microbiome showed similar microbial diversity to the control, independent of antiviral treatment. Decreased Bacteroidota and increased Firmicutes and Proteobacteria were observed in the lungs in a virus dose-dependent manner. These studies add to a better understanding of the complexities associated with the intestinal microbiome during respiratory infections.

microbiology↗

Rapid and stable microbial community assembly in the headwaters of third-order stream

Small streams and their headwaters are a key source of microbial diversity in fluvial systems and serve as an entry point for bacteria from the surrounding landscape. Community assembly processes occurring in these streams shape downstream population structure and nutrient cycles. To elucidate the development and stability of microbial communities along the length of a first through third order stream, fine-scale temporal and spatial sampling regimes were employed along McNutt Creek in Athens, Georgia, USA. 16S rRNA gene libraries were constructed from samples collected on a single day from 19 sites spanning the first 16.76 km of the stream. Selected sites at the upper, mid, and lower reaches of the stream were sampled daily for 11 days to evaluate community variability over time. In a second study, sites at and near the creeks headwaters were sampled daily for 11 days to understand the initial stages of bacterioplankton community assembly. In all studies, we observed decreasing alpha and beta diversity with increasing downstream distance. These trends were accompanied by the enrichment of a small fraction of taxa found at low abundance in the furthest-upstream environments. Similar sets of taxa consistently increased significantly in relative abundance in downstream samples over time scales ranging from 1 day to 1 year, many of which belong to microbial clades known to be abundant in freshwater environments. These results underpin the importance of headwaters as the site of rapid in-stream selection that results in the reproducible establishment of a highly stable community of freshwater riverine bacteria.\n\nImportanceHeadwater streams are critical introduction points of microbial diversity for larger connecting rivers and play key roles in the establishment of taxa that partake in in-stream nutrient cycling. We examined microbial community composition of a first- through third-order stream using fine-scale temporal and spatial regimes. Our results show that the bacterioplankton community develops rapidly and predictably from the headwater population with increasing total stream length. Along the length of the stream, the microbial community exhibits substantial diversity loss and enriches repeatedly for select taxa across days and years, although the relative abundances of individual taxa vary over time and space. This repeated enrichment of a stable stream community likely contributes to the stability and flexibility of downstream communities.

microbiology↗