Microorganisms of Two Thermal Pools on Kunashir Island, Russia

The Kuril Archipelago is a part of the Circum-Pacific Belt (Ring of Fire). These islands have numerous thermal springs. There are very few studies on these microbial communities, and none of them have been conducted by modern molecular biological methods. Here we performed the first metagenomic study on two thermophilic microbial communities of Kunashir Island. Faust Lake is hot (48 °C) and highly acidic (pH 2.0). We constructed 28 metagenome-assembled genomes as well as 17 16S ribosomal RNA sequences. We found that bottom sediments of Faust Lake are dominated by a single species of red algae belonging to the Cyanidiaceae family. Archaeans in Faust Lake are more diverse than bacteria but less abundant. The Tretyakovsky Thermal Spring is also hot (52 °C) but only weakly acidic (pH 6.0). It has much higher microbial diversity (233 metagenome-assembled genomes; 93 16S ribosomal RNAs) and is dominated by bacteria, with only several archaeans and one fungus. Despite their geographic proximity, these two thermal springs were found to not share any species. A comparison of these two lakes with other thermal springs of the Circum-Pacific Belt revealed that only a few members of the communities are shared among different locations.

Stony Coral Tissue Loss Disease biomarker bacteria identified in corals and overlying waters using a rapid field-based sequencing approach

Stony Coral Tissue Loss Disease (SCTLD) is a devastating disease. Since 2014, it has spread along the entire Florida Reef Tract, presumably via a water-borne vector, and into the greater Caribbean. It was first detected in the United States Virgin Islands (USVI) in January 2019. To more quickly identify disease biomarker microbes, we developed a rapid pipeline for microbiome sequencing. Over a span of 10 days we collected, processed, and sequenced coral tissue and near-coral seawater microbiomes from diseased and apparently healthy Colpophyllia natans, Montastraea cavernosa, Meandrina meandrites and Orbicella franksi. Analysis of the resulting bacterial and archaeal 16S ribosomal RNA sequences revealed 25 biomarker amplicon sequence variants (ASVs) enriched in diseased tissue. These biomarker ASVs were additionally recovered in near-coral seawater (within 5 cm of coral surface), a potential recruitment zone for pathogens. Phylogenetic analysis of the biomarker ASVs belonging to Vibrio, Arcobacter, Rhizobiaceae, and Rhodobacteraceae revealed relatedness to other coral disease-associated bacteria and lineages novel to corals. Additionally, four ASVs (Algicola, Cohaesibacter, Thalassobius and Vibrio) were exact sequence matches to microbes previously associated with SCTLD. This work represents the first rapid coral disease sequencing effort and identifies biomarkers of SCTLD that could be targets for future SCTLD research.

Pigmented Pseudoalteromonas Sp. Isolated from Marine Sponge with Anti-Microbial Activities against Selected Human Pathogens

Marine sponges have been the potential source of bioactive compounds with potent antimicrobial properties. Sponge associated microbes significantly provide the route of biosynthesis of some of these compounds. In this work, a total of 100 bacterial colonies were screened from a marine sponge from Class Demospongiae, which has been collected from Merambong Island, the state of Johor, Malaysia. In disk diffusion assay, only 2 out of 100 isolates; namely C40 and C52, were able to demonstrate active inhibitions against selected human pathogens (Pseudomonas aeruginosa, Escherichia coli, Bacillus subtilis, except for Staphylococcus aureus). Isolates C40 and C52 were characterized to be Gram negative short rods, non-spore formers and catalase positive. Unlike the majority of other isolates from sponge which were Gram positive rods, Isolate C40 and C52 are Gram negative rods which grew in yellow pigmented colonies. Genotypic characterization using 16S ribosomal RNA sequencing were carried out on each isolate (accession number for C40 and C52 is MT645493 and MT645494, respectively). The 16S ribosomal RNA sequences revealed that these strains belonged to genus Pseudoalteromonas sp. with 97-98% similarities. Inhibitions studies showed that this sponge associated microorganisms potentially produce anti-microbial compounds useful for biotechnologies.

Origins, History and Molecular Characterization of Creole Cane

Since it was first introduced to Europe in 711 CE and planted in the Americas in 1506, a single type of cane dominated sugar production for 1100 years, until it was finally ousted by Tahitian cane c. 1790. This cane became known as ‘CreoleâĂŹ and is present in the ancestry of many sugarcane hybrids, even today. Whether there was only a single variety of Creole cane or multiple varieties has been a matter of debate for decades. Creole cane remains relevant today, as a Creole cane from Jamaica is the currently chosen lecotype for Saccharum officinarum. In this study we identify 18 historical images of Creole cane, many not previously published. We employ image analyses to characterize the internodes and demonstrate evidence for only a single type of Creole in the new world. Chloroplasts and 45s ribosomal RNA sequences from the cultivar BH10/12 (known to have a Creole female parent) were determined that Java ribbon cane is the historical New World sugarcane known as Creole. We demonstrate that Creole cane is an hybrid and not a single species. Thus S. officinarum has no type specimen. We also sequence a ribbon cane (also known as Guinguam) that appeared in the Caribbean between 1790 and 1810 and demonstrate that this cane was a Sinense type from Java that links back to the work of Rumphinus (1660s).

Nuclease-mediated depletion biases in ribosome footprint profiling libraries

Ribosome footprint profiling is a high throughput sequencing based technique that provides detailed and global views of translation in living cells. An essential part of this technology is removal of unwanted, normally very abundant, ribosomal RNA sequences that dominate libraries and increase sequencing costs. The most effective commercial solution (Ribo-Zero) has been discontinued and a number of new, experimentally distinct commercial applications have emerged on the market. Here we evaluated several commercially available alternatives designed for RNA-seq of human samples and find them unsuitable for ribosome footprint profiling. We instead recommend the use of custom-designed biotinylated oligos, which were widely used in early ribosome profiling studies. Importantly, we warn that depletion solutions based on targeted nuclease cleavage significantly perturb the high-resolution information that can be derived from the data, and thus do not recommend their use for any applications that require precise determination of the ends of RNA fragments.

Molecular phylogeny of historical micro-invertebrate specimens using de novo sequence assembly

Resolution of relationships at lower taxonomic levels is crucial for answering many evolutionary questions, and as such, sufficiently varied species representation is vital. This latter goal is not always achievable with relatively fresh samples. To alleviate the difficulties in procuring rarer taxa, we have seen increasing utilization of historical specimens in building molecular phylogenies using high throughput sequencing. This effort, however, has mainly focused on large-bodied or well-studied groups, with small-bodied and under-studied taxa under-prioritized. Here, we present a pipeline that utilizes both historical and contemporary specimens, to increase the resolution of phylogenetic relationships among understudied and small-bodied metazoans, namely, cheilostome bryozoans. In this study, we pioneer sequencing of air-dried bryozoans, utilizing a recent library preparation method for low DNA input. We use the de novo mitogenome assembly from the target specimen itself as reference for iterative mapping, and the comparison thereof. In doing so, we present mitochondrial and ribosomal RNA sequences of 43 cheilostomes representing 37 species, including 14 from historical samples ranging from 50 to 149 years old. The inferred phylogenetic relationships of these samples, analyzed together with publicly available sequence data, are shown in a statistically well-supported 65 taxa and 17 genes cheilostome tree. Finally, the methodological success is emphasized by circularizing a total of 27 mitogenomes, seven from historical cheilostome samples. Our study highlights the potential of utilizing DNA from micro-invertebrate specimens stored in natural history collections for resolving phylogenetic relationships between species.

PhySpeTree: an automated pipeline for reconstructing phylogenetic species trees

Background Phylogenetic species trees are widely used in inferring evolutionary relationships. Existing software and algorithms mainly focus on phylogenetic inference. However, less attention has been paid to intermediate steps, such as processing extremely large sequences and preparing configure files to connect multiple software. When the species number is large, the intermediate steps become a bottleneck that may seriously affect the efficiency of tree building. Results Here, we present an easy-to-use pipeline named PhySpeTree to facilitate the reconstruction of species trees across bacterial, archaeal, and eukaryotic organisms. Users need only to input the abbreviations of species names; PhySpeTree prepares complex configure files for different software, then automatically downloads genomic data, cleans sequences, and builds trees. PhySpeTree allows users to perform critical steps such as sequence alignment and tree construction by adjusting advanced options. PhySpeTree provides two parallel pipelines based on concatenated highly conserved proteins and small subunit ribosomal RNA sequences, respectively. Accessory modules, such as those for inserting new species, generating visualization configurations, and combining trees, are distributed along with PhySpeTree. Conclusions Together with accessory modules, PhySpeTree significantly simplifies tree reconstruction. PhySpeTree is implemented in Python running on modern operating systems (Linux, macOS, and Windows). The source code is freely available with detailed documentation (https://github.com/yangfangs/physpetools).

Exploring the genetic diversity of the 16S rRNA gene of Akkermansia muciniphila in IBD and IBS

Aim: The human gastrointestinal tract harbors diverse, abundant microbiota and Akkermansia muciniphila is involved in this community. The aim of this study is to characterize 16 new A. muciniphila 16S ribosomal RNA sequences selected from a metagenomic database from stools of patients with irritable bowel syndrome (IBS), inflammatory bowel diseases and control (CTRLs) subjects by a phylogenetic approach. Materials & methods: A phylogenetic approach was used to study the genetic diversity and SNPs in 16 A. muciniphila 16S ribosomal RNA sequences from stools of 107 individuals, 36 of which were patients affected by IBS, 30 by inflammatory bowel disease and 41 were CTRLs. Results: Phylogenetic analysis confirmed the subdivision into different supported clusters. An increase of variability in IBS has been identified. Conclusion: The genetic variation combined to the relative abundance, contribute to the protective role of A. muciniphila. Phylogenesis represent an additional approach to investigate genetic variability.

Identification of the new species Comatricha macrospora and two other recently recorded species of Comatricha from China

A new species (Comatricha macrospora) has been collected from the Changbai Mountain National Nature Reserve, Jilin Province, China. Comatricha macrospora has smaller sporocarps and larger spores (about 15–18 μm in diameter) than other species of Comatricha as well as a persistent peridium at the base of the sporotheca. In addition, two newly recorded species of Comatricha—C. tenerrima (M.A. Curtis) G. Lister and C. afroalpina Rammeloo—have been documented in China for the first time, based on material collected from northeast China and the campus of East China Normal University, Fuzhou City, Jiangxi Province. Comatricha tenerrima is characterised by fusiform long-stalked sporocarps and warted pinkish brown spores (about 7–8 μm in diameter)., whereas C. afroalpina occurs on rotting logs and has spores marked by larger warts with an irregular reticulation at their base. Descriptions and scanning electron micrographs for these members of the genus Comatricha are provided. Phylogenetic analyses, based on small subunit ribosomal RNA sequences (SSUrRNA) of Comatricha and related genera, were carried out using Bayesian inference. These analyses confirmed the placement of the new species in the genus Comatricha.