Omics Approaches to Assess Flavor Development in Cheese

Cheese is characterized by a rich and complex microbiota that plays a vital role during both production and ripening, contributing significantly to the safety, quality, and sensory characteristics of the final product. In this context, it is vital to explore the microbiota composition and understand its dynamics and evolution during cheese manufacturing and ripening. Application of high-throughput DNA sequencing technologies have facilitated the more accurate identification of the cheese microbiome, detailed study of its potential functionality, and its contribution to the development of specific organoleptic properties. These technologies include amplicon sequencing, whole-metagenome shotgun sequencing, metatranscriptomics, and, most recently, metabolomics. In recent years, however, the application of multiple meta-omics approaches along with data integration analysis, which was enabled by advanced computational and bioinformatics tools, paved the way to better comprehension of the cheese ripening process, revealing significant associations between the cheese microbiota and metabolites, as well as their impact on cheese flavor and quality.

Efficiency and bacterial diversity of an improved anaerobic baffled reactor for the remediation of wastewater from alkaline-surfactant-polymer (ASP) flooding technology

Alkaline-surfactant-polymer (ASP) flooding technology is used to maximize crude oil recovery. However, the extensive use of alkaline materials makes it difficult to treat the water used. Here, an improved multi-zone anaerobic baffled reactor (ABR) using FeSO4 as electron acceptor was employed to treat the wastewater from ASP flooding technology, and the effects on major pollutants (hydrolyzed polyacrylamide, petroleum substances, surfactants suspended solids) and associated parameters (chemical oxygen demand, viscosity) were evaluated. Gas chromatography-mass spectrometry (GC-MS) was used to follow the degradation and evolution of organic compounds while high-throughput DNA sequencing was used to determine the bacterial diversity in the ABR. The results obtained after 90 d of operation showed decreases in all parameters measured and the highest mean removal rates were obtained for petroleum substances (98.8%) and suspended solids (77.0%). Amounts of petroleum substances in the ABR effluent could meet the requirements of a national standard for oilfield reinjection water. GC-MS analysis showed that a wide range of chemicals (e.g. aromatic hydrocarbons, esters, alcohols, ketones) could be sequentially removed from the influent by each zone of ABR. The high-throughput DNA sequencing showed that the bacteria Micropruina, Saccharibacteria and Synergistaceae were involved in the degradation of pollutants in the anaerobic and anoxic reaction zones, while Rhodobacteraceae and Aliihoeflea were the main functional microorganisms in the aerobic reaction zones. The results demonstrated that the improved ABR reactor had the potential for the treatment of wastewater from ASP flooding technology.

Characteristics of the Fungal Communities and Co-occurrence Networks in Hazelnut Tree Root Endospheres and Rhizosphere Soil

Hazelnut has gained economic value in China in recent years, but its large-scale planting and research started later than other countries. Conducting basic research on hazelnut trees requires studying their related microorganisms. Here, we used high-throughput DNA sequencing to quantify the fungal communities in the root endospheres and rhizosphere soil of four hazelnut species. Fungal diversity in the rhizosphere soil was significantly higher than that in the root endospheres. Rhizosphere soil had more Mortierellomycota, and the fungal community compositions differed among the four hazelnut species. The root endospheres, especially those of the Ping’ou (Corylus heterophylla × Corylus avellana) trees, contained more ectomycorrhizal fungi. The co-occurrence networks in the rhizosphere soil were more sophisticated and stable than those in the root endospheres, even when the root endospheres had higher modularity, because the structural differentiation of the root endospheres differed from that of the rhizosphere soil. Two-factor correlation network analysis and linear regression analysis showed that the total organic carbon was the main environmental factor affecting the fungal communities. Our study revealed the community compositions, functional predictions, and co-occurrence network structural characteristics of fungi in hazelnut root endospheres and rhizosphere soil. We also examined the potential keystone taxa, and analyzed the environmental factors of the dominant fungal community compositions. This study provides guidance for the growth of hazelnut and the management of hazelnut garden, and provides an insight for future development of fungal inoculants to be used in hazelnut root.

Soil Fungal Diversity of the Aguarongo Andean Forest (Ecuador)

Fungi represent an essential component of ecosystems, functioning as decomposers and biotrophs, and they are one of the most diverse groups of Eukarya. In the tropics, many species are unknown. In this work, high-throughput DNA sequencing was used to discover the biodiversity of soil fungi in the Aguarongo forest reserve, one of the richest biodiversity hotspots in Ecuador. The rDNA metabarcoding analysis revealed the presence of seven phyla: Ascomycota, Basidiomycota, Mortierellomycota, Mucoromycota, Glomeromycota, Chytridiomycota, and Monoblepharomycota. A total of 440 identified species were recorded. They mainly belonged to Ascomycota (263) and Basidiomycota (127). In Mortierellomycota, 12 species were recorded, among which Podila verticillata is extremely frequent and represents the dominant species in the entire mycobiota of Aguarongo. The present research provides the first account of the entire soil mycobiota in the Aguarongo forest, where many fungal species exist that have strong application potential in agriculture, bioremediation, chemical, and the food industry. The Aguarongo forest hides a huge number of unknown fungal species that could be assessed, and its protection is of the utmost importance.

Placental Microbiome: a Paradigm Shift or Flaw in the Methodology?

Rapid development of high-throughput DNA sequencing technologies and bioinformatics methods, together with a substantial reduction of their cost, have provided tremendous opportunities for studying the human microbiome. In recent years, much attention has been paid to studies of the microbiome of the upper reproductive tract of woman and the fetoplacental system, which have traditionally been considered sterile. Obtaining irrefutable evidence of the existence of the placental microbiome would enable us to believe with a high degree of certainty that microorganisms colonize the fetus already in the womb, which would have far-reaching consequences not only for medicine, but also for basic biology. This issue triggered a heated discussion among microbiologists, molecular biologists, obstetricians, and neonatologists. In the past few years, a number of studies have been published, both refuting and confirming the dogma, accepted for many decades, that the placenta and fetus are sterile during a healthy pregnancy. This literature review is a critical analysis of the results of studies into the placental microbiome. It provides arguments both for supporters of the hypothesis of the resident microbiota of the placenta and their opponents. Particular attention is paid to the methodological requirements for molecular studies of biological material with low microbial biomass, compliance with which is crucial for obtaining reliable results.

Functional DNA annotation from a preliminary de novo genome assembly of Brycon orbignyanus, an endangered Neotropical migratory fish

The predicted sequence for thousands of genes revealed by a preliminary low-coverage genome assembly is presented for Brycon orbignyanus, an endangered migratory fish. Neotropical migratory fish stocks have been drastically reduced due to accumulated environmental pressure. Brycon orbignyanus, once one of the main fisheries species in the Platine Basin, is now very rare in nature and relies on spawning programs and a few well preserved or still untouched sites. The use of high-throughput DNA sequencing is still untapped regarding the functional genome information from B. orbignyanus. In order to help bridging this gap, we present a dataset resulting from the first functional annotation from a de novo genome assembly for B. orbignyanus, from short reads (90 bp), obtained by the HiSeq 2000 platform (Illumina). The annotation was performed for scaffolds over 10 kb using the Maker pipeline, with reference sequences taken from the NCBI for the Characiformes order. This annotation resulted in the prediction of 12,734 genes, classified with the aid of PANTHER. The data presented here can facilitate the development of basic research in this threatened species, along with practical biotechnological tools for different areas, such as commercial and environmental fish spawning operations (e.g. hormonal induction, growth) and human health.

Rhizosphere Microbiomes of Potato Cultivated under Bacillus subtilis Treatment Influence the Quality of Potato Tubers

Plants serve as a niche for the growth and proliferation of a diversity of microorganisms. Soil microorganisms, which closely interact with plants, are increasingly being recognized as factors important to plant health. In this study, we explored the use of high-throughput DNA sequencing of the fungal ITS and bacterial 16S for characterization of the fungal and bacterial microbiomes following biocontrol treatment (DT) with Bacillus subtilis strain Bv17 relative to treatments without biocontrol (DC) during the potato growth cycle at three time points. A total of 5631 operational taxonomic units (OTUs) were identified from the 16S data, and 2236 OTUs were identified from the ITS data. The number of bacterial and fungal OTU in DT was higher than in DC and gradually increased during potato growth. In addition, indices such as Ace, Chao, Shannon, and Simpson were higher in DT than in DC, indicating greater richness and community diversity in soil following the biocontrol treatment. Additionally, the potato tuber yields improved without a measurable change in the bacterial communities following the B. subtilis strain Bv17 treatment. These results suggest that soil microbial communities in the rhizosphere are differentially affected by the biocontrol treatment while improving potato yield, providing a strong basis for biocontrol utilization in crop production.

Molecular recording of sequential cellular events into DNA

Genetically encoded DNA recorders noninvasively convert transient biological events into durable mutations in a cell's genome, allowing for the later reconstruction of cellular experiences using high-throughput DNA sequencing. Existing DNA recorders have achieved high-information recording, durable recording, prolonged recording over multiple timescales, multiplexed recording of several user-selected signals, and temporally resolved signal recording, but not all at the same time. We present a DNA recorder called peCHYRON (prime editing Cell HistorY Recording by Ordered iNsertion) that does. In peCHYRON, prime editor guide RNAs (pegRNAs) insert a variable triplet DNA sequence alongside a constant propagation sequence that deactivates the previous and activates the next step of insertion. This process results in the sequential accumulation of regularly spaced insertion mutations at a synthetic locus. Accumulated insertions are permanent throughout editing because peCHYRON uses a prime editor that avoids cutting both DNA strands, which risks deletions. Editing continues indefinitely because each insertion adds the complete sequence needed to initiate the next step. Constitutively expressed pegRNAs generate insertion patterns that support straightforward reconstruction of cell lineage relationships. Pulsed expression of different pegRNAs enables the reconstruction of pulse sequences, which may be coupled to biological stimuli for temporally-resolved multiplexed event recording.

Tet1 Regulates Astrocyte Development and Cognition of Mice Through Modulating GluA1

Tet (Ten eleven translocation) family proteins-mediated 5-hydroxymethylcytosine (5hmC) is highly enriched in the neuronal system, and is involved in diverse biological processes and diseases. However, the function of 5hmC in astrocyte remains completely unknown. In the present study, we show that Tet1 deficiency alters astrocyte morphology and impairs neuronal function. Specific deletion of Tet1 in astrocyte impairs learning and memory ability of mice. Using 5hmC high-throughput DNA sequencing and RNA sequencing, we present the distribution of 5hmC among genomic features in astrocyte and show that Tet1 deficiency induces differentially hydroxymethylated regions (DhMRs) and alters gene expression. Mechanistically, we found that Tet1 deficiency leads to the abnormal Ca2+ signaling by regulating the expression of GluA1, which can be rescued by ectopic GluA1. Collectively, our findings suggest that Tet1 plays important function in astrocyte physiology by regulating Ca2+ signaling.