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Neobacillus driksii sp. nov. isolated from a Mars 2020 spacecraft assembly facility and genomic potential for lasso peptide production in Neobacillus.

UNLABELLED: During microbial surveillance of the Mars 2020 spacecraft assembly facility, two novel bacterial strains, potentially capable of producing lasso peptides, were identified. Characterization using a polyphasic taxonomic approach, whole-genome sequencing and phylogenomic analyses revealed a close genetic relationship among two strains from Mars 2020 cleanroom floors (179-C4-2-HS, 179-J1A1-HS), one strain from the Agave plant (AT2.8), and another strain from wheat-associated soil (V4I25). All four strains exhibited high 16S rRNA gene sequence similarity (>99.2%) and low average nucleotide identity (ANI) with Neobacillus niacini NBRC 15566T, delineating new phylogenetic branches within the genus. Detailed molecular analyses, including gyrB (90.2%), ANI (86.4%), average amino acid identity (87.8%) phylogenies, digital DNA-DNA hybridization (32.6%), and percentage of conserved proteins (77.7%) indicated significant divergence from N. niacini NBRC 15566T. Consequently, these strains have been designated Neobacillus driksii sp. nov., with the type strain 179-C4-2-HST (DSM 115941T = NRRL B-65665T). N. driksii grew at 4°C to 45°C, pH range of 6.0 to 9.5, and 0.5% to 5% NaCl. The major cellular fatty acids are iso-C15:0 and anteiso-C15:0. The dominant polar lipids include diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and an unidentified aminolipid. Metagenomic analysis within NASA cleanrooms revealed that N. driksii is scarce (17 out of 236 samples). Genes encoding the biosynthesis pathway for lasso peptides were identified in all N. driksii strains and are not commonly found in other Neobacillus species, except in 7 out of 26 recognized species. This study highlights the unique metabolic capabilities of N. driksii, underscoring their potential in antimicrobial research and biotechnology. IMPORTANCE: The microbial surveillance of the Mars 2020 assembly cleanroom led to the isolation of novel N. driksii with potential applications in cleanroom environments, such as hospitals, pharmaceuticals, semiconductors, and aeronautical industries. N. driksii genomes were found to possess genes responsible for producing lasso peptides, which are crucial for antimicrobial defense, communication, and enzyme inhibition. Isolation of N. driksii from cleanrooms, Agave plants, and dryland wheat soils, suggested niche-specific ecology and resilience under various environmentally challenging conditions. The discovery of potent antimicrobial agents from novel N. driksii underscores the importance of genome mining and the isolation of rare microorganisms. Bioactive gene clusters potentially producing nicotianamine-like siderophores were found in N. driksii genomes. These siderophores can be used for bioremediation to remove heavy metals from contaminated environments, promote plant growth by aiding iron uptake in agriculture, and treat iron overload conditions in medical applications.

Phylogeny↗

PCR Sampling of disease resistance-like sequences from a disease resistance gene cluster in soybean.

Clusters of Resistance-like genes (RLGs) have been identified from a variety of plant species. In soybean, RLG-specific primers and BAC-fingerprinting were used to develop a contig of overlapping BACs for a cluster of RLGs on soybean linkage group J. The resistance genes Rps2 ( Phytophthora stem and root rot) and Rmd-c (powdery mildew) and the ineffective nodulation gene Rj2 were previously mapped to this region of linkage group J. PCR hybridization was used to place two TIR/NBD/LRR cDNAs on overlapping BACs from this contig. Both of the cDNAs were present on BAC 34P7. Fingerprinting of this BAC suggested as many as twelve different RLGs were present. Given the high nucleotide identity shared between cDNAs LM6 and MG13 (>90%), direct sequencing of this region would be difficult. More sequence information was needed about the RLGs within this region before sequencing could be undertaken. By comparing the genomic sequences of cDNAs LM6 and MG13 we identified conserved regions from which oligonucleotide primers specific to BAC 34P7 RLGs could be designed. The nine primer pairs spanned the genomic sequence of LM6 and produced overlapping RLG products upon amplification of BAC 34P7. Amplification products from 12 different RLGs were identified. On average, nucleotide identity between RLG sequences was greater than 95%. Examination of RLG sequences also revealed evidence of additions, deletions and duplications within targeted regions of these genes. Using previously mapped cDNAs we were able to quickly and inexpensively access multiple RLGs within a single specific cluster.

Journal Article↗

Streptomyces huangiella sp. nov., an endophytic actinomycete isolated from Pheretima aspergillum, a promising candidate for biological pathogen control.

UNLABELLED: Pheretima aspergillum (E. Perrier) is an annelid of the genus Pheretima in the family Megascolecidae, a species of earthworm, whose dried body (Guang Dilong) is a traditional Chinese animal medicine. A new actinobacterium strain, named HD1123-B1T, was isolated from the gut contents of Pheretima aspergillum caught in the wild in Guangzhou, China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strain was primarily identified as a member of the genus Streptomyces, sharing more than 98% sequence identity to Streptomyces endocoffeicus CA3R110T (98.80%), Streptomyces coffeae CA1R205T (98.47%), and Streptomyces iranensis HM35T (97.93%). The whole genome size of strain HD1123-B1T was approximately 8.9 Mbp, with 7,464 predicted genes and 71.42 mol% DNA C+G content. Comparative genomic analyses based on digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values revealed that strain HD1123-B1T represents a novel species within the genus Streptomyces. Additionally, 38 biosynthetic gene clusters for secondary metabolites were also predicted in the genome of strain HD1123-B1T. Based on LC-MS/MS analysis, the nigericin biosynthesis gene cluster has been completely characterized. The ethyl acetate crude extract of strain HD1123-B1T exhibited remarkable antibacterial activity against gram-positive bacteria (methicillin-resistant Staphylococcus aureus ATCC 25213, etc) and gram-negative bacteria Ralstonia solanacearum GIM 1.70. Based on these results, HD1123-B1T could be confirmed as an isolate that represents a novel species of the genus Streptomyces, for which the name Streptomyces huangiella sp. nov. is proposed. IMPORTANCE: As the largest genus of the phylum Actinomycetes, Streptomyces is a kind of microbial resources with great practical and economic value. Due to their unique physiological properties and metabolic capacity, Streptomyces have become an important source of bioactive compounds in the world and play an indispensable role in medical and industrial fields. With the advancement of molecular biology and genomics, researchers can more deeply explore the metabolic potential of Actinomycetes, discovering and developing new biologically active compounds. These new compounds may possess various biological activities, such as antibacterial, antiviral, antifungal, and antiparasitic properties, further promoting the development of medicine and related industries. Based on genomic analysis and antibacterial activity, the strain HD1123-B1T was indicated to be a promising candidate for biological pathogen control.

Streptomyces↗

Characterization of Dapalides D and E and Genomic Comparison of the Two Co-Occurring Dapalide-Producing Dapis spp.

Marine cyanobacteria are a rich source of diverse bioactive natural products, targeting proteins involved in many diseases. Here, we combined metagenomic analysis to enhance the structure elucidation process of two new cyclodepsipeptides named dapalides D (1) and E (2) from a collection of a cyanobacterial mat containing multiple Dapis species from Guam. Dapalides D/E are composed of 11 amino acids, including multiple identical units with different configurations. Enantioselective amino acid identification of the acid hydrolyzate established the identity of amino acids, including the configuration of α/β-stereogenic centers. Identification and analysis of the dapalides D/E biosynthetic gene cluster from a metagenome-assembled genome aided the elucidation of α-configuration and establishment of the order of individual building blocks, collectively revealing the total structure. Phylogenomic analysis indicates that the dapalides D/E producer belongs to Dapis sp. (Dapis sp. VPG23-80 MAG-2), which shares a 95.2% average nucleotide identity with Dapis sp. VPG23-80 MAG-1, the producer of dapalides A-C that cooccurs in the same assemblage. Dapalide D (1) showed moderate growth inhibitory activity against various cancer cell lines. This work expands the dapalide structure class and further highlights the use of combined chemical and metagenomic analyses for natural product structure elucidation.

Cyanobacteria↗

Isolation and genomic characterization of Bacillus X32: a potent phosphate-solubilizing bacterium with growth-promoting effects on navel orange seedlings.

Phosphorus is an essential element for plant growth. However, in nature, most phosphorus exists in the form of insoluble compounds that plants cannot directly absorb, leading to phosphorus deficiency in agricultural systems. With increasing demand for economic crops such as citrus and the decline in soil fertility due to current management practices, there is a growing need for environmentally friendly fertilizers to improve and restore soil conditions. In this study, a highly efficient phosphate‑solubilizing strain X32 was isolated from the rhizosphere soil of Gannan navel oranges. Systematic genomic analysis identified it as a putative novel species within the genus Bacillus, showing the closest phylogenetic relationship to Bacillus spizizenii. However, both the average nucleotide identity (ANI = 93.18%) and digital DNA‑DNA hybridization (dDDH = 50.4%) values fell below the established thresholds for species delineation, indicating significant genomic differentiation. Whole‑genome sequencing further revealed that strain X32 harbors multiple functional genes potentially related to phosphorus metabolism, including inorganic phosphate‑solubilizing genes (e.g., gdh and gltA), phosphate transport genes (e.g., glpT, pstA, pstB, pstC), and phosphorus mineralization genes (e.g., phoA, phoD). Pot experiment results demonstrated that inoculation with strain X32 significantly promoted the growth of navel orange seedlings, as evidenced by marked increases in both aboveground and belowground fresh and dry weights, as well as plant height. Additionally, strain X32 significantly enhanced the activities of antioxidant enzymes (SOD, CAT, POD) and regulated the content of chlorophyll b in seedling leaves, these changes suggest that strain X32 may enhance stress resistance in plants and influence photosynthetic pigment composition, though direct measurements of photosynthetic performance are needed for confirmation. This study provides a theoretical basis for developing microbial fertilizers with efficient phosphorus solubilization and plant growth-promoting functions, which may help reduce dependence on phosphorus fertilizers and promote sustainable agricultural development.

Phosphates↗

Pseudomonas gebzensis sp. nov., a Carotenoid-Producing Species Isolated from Solar Panels in Türkiye.

Strain RP23018ST was isolated from the solar panels in Gebze, Türkiye. According to the phylogenetic analysis of 16S rRNA gene sequences, strain RP23018ST was classified as a member of the genus Pseudomonas, being most closely related to Pseudomonas cremoricolorata DSM 17059T (99.3%). Strain RP23018ST was Gram-stain-negative, rod-shaped, aerobic, motile, growing between temperature range of 4–35 °C. The polar lipids of strain RP23018ST consisted of phosphatidylethanolamine, di-phosphatidylglycerol, phosphatidylcholine, one unidentified phospholipid, and three unidentified aminolipids. It was determined that the main respiratory quinone of the RP23018ST strain was ubiquinone-9 (Q-9), which is a typical chemotaxonomic characteristic of the genus Pseudomonas. Major fatty acids were C16:0, summed feature 3 (C16:1 ω6c and/or C16:1 ω7c), summed feature 8 (C18:1 ω6c and/or C18:1 ω7c), and C17:0 cyclo. Strain RP23018ST had a genome size of 4,944,510 bp and a G + C ratio of 63.0%. The average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values between RP23018ᵀ and related Pseudomonas species were below 86.4% and 26.7%, respectively, clearly falling below the thresholds for species delineation. Strain RP23018ST produced a deep yellow carotenoid pigment with absorption maxima at 455 and 482 nm. Genome annotation revealed key carotenoid biosynthesis genes (crtX, crtY, crtI, crtZ), and the extracted pigment showed antioxidant activity in DPPH and FRAP assays. Polyphasic characterization, including the determination of overall genome relatedness indices, revealed that strain RP23018ST represents a novel species of the genus Pseudomonas, for which the name Pseudomonas gebzensis sp. nov. is proposed. The type strain is RP23018ST (= LMG 33436T = DSM 117295T).

Turkey↗

Curtobacterium uvaldense sp. nov., Falsirhodobacter allii sp. nov., Pseudomonas texasensis sp. nov., Three Novel Species Isolated from Onions in Texas, USA.

Bacterial strains 20TX0008T and 20TX0035T were isolated from the symptomatic onion foliage in 2020, and a bacterial strain 21TX0197T was isolated from the symptomatic onion bulb in 2021. Pathogenicity assays confirmed the two strains isolated from foliage to be non-pathogenic, while the strain 21TX0197T was found to be pathogenic. Phylogenetic analysis using the 16&#xa0;S rRNA gene revealed the closest resemblance of 20TX0008T, 20TX0035T, and 21TX0197T to the genera Curtobacterium, Falsirhodobacter, and Pseudomonas, respectively. For the respective genus, strains 20TX0008T showed the greatest 16&#xa0;S rRNA sequence similarity with Curtobacterium caesinilyticum GDMCC 1.2667T (99.7%), 20TX0035T with Falsirhodobacter deserti W402T (99.1%), and 21TX0197T with Pseudomonas uvaldensis 20TX0172T (99.4%). In the phylogenetic trees based on whole genome sequences, all three strains formed a distinct, well-supported branch that separated them from their closely related species within the same genus. The genomic similarity, based on average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH), revealed that the novel strains had values below the species delineation thresholds (ANI&#x2009;<&#x2009;95% and dDDH&#x2009;<&#x2009;70%), confirming the novelty of all these strains. Based on genomic and phenotypic characteristics, we propose three novel species, Curtobacterium uvaldense sp. nov. (type strain 20TX0008T =LMG 33207T =NCIMB 15480T), Falsirhodobacter allii sp. nov. (type strain 20TX0035T =LMG 33208T =NCIMB 15477T), and Pseudomonas texasensis sp. nov. (type strain 21TX0197T =LMG 33210T =NCIMB 15479T).

Phylogeny↗

Duganella hordei sp. nov., Duganella caerulea sp. nov., and Duganella rhizosphaerae sp. nov., isolated from barley rhizosphere.

Duganella sp. strains R1T, R57T, and R64T, isolated from barley roots in Japan, are Gram-stain-negative, motile, rod-shaped bacteria. Duganella species abundantly colonized barley roots. Strains R1T, R57T, and R64T were capable of growth at 4&#xa0;&#xb0;C, suggesting adaptation to colonize winter barley roots. Strains R57T and R64T formed purple colonies, indicating violacein production, while strain R1T did not. Based on 16S rRNA gene sequence similarities, strains R1T, R57T, and R64T were most closely related to D. violaceipulchra HSC-15S17T (99.10%), D. vulcania FT81WT (99.45%), and D. violaceipulchra HSC-15S17T (99.86%), respectively. Their genome sizes ranged from 7.05 to 7.38 Mbp, and their genomic G+C contents were 64.2-64.7%. The average nucleotide identity and digital DNA-DNA hybridization values between R1T and D. violaceipulchra HSC-15S17T, R57T and D. vulcania FT81WT, R64T and D. violaceipulchra HSC-15S17T were 86.0% and 33.2%, 95.7% and 67.9%, and 92.7% and 52.6%, respectively. Their fatty acids were predominantly composed of C16:0, C17:0 cyclo, and summed feature 3 (C16:1 &#x3c9;7c and/or C16:1 &#x3c9;6c). Based on their distinct genetic and phenotypic characteristics, and supported by chemotaxonomic analyses, we propose that strains R1T, R57T, and R64T represent novel species within the Duganella genus, for which the names Duganella hordei (type strain R1T&#x2009;=&#x2009;NBRC 115982&#xa0;T&#x2009;=&#x2009;DSM 115069&#xa0;T), Duganella caerulea (type strain R57T&#x2009;=&#x2009;NBRC 115983&#xa0;T&#x2009;=&#x2009;DSM 115070&#xa0;T), and Duganella rhizosphaerae (type strain R64T&#x2009;=&#x2009;NBRC 115984&#xa0;T&#x2009;=&#x2009;DSM 115071&#xa0;T) are proposed.

Hordeum↗

Genomic and polyphasic characterization of six novel Hymenobacter species isolated from soil in Korea.

Six bacterial strains (BT523T, BT559T, 15J16-1T3BT, BT730T, DG25AT, and DG25BT) were isolated from soil samples in Korea and assigned to the family Hymenobacteraceae (order Cytophagales, class Cytophagia). Phylogenetic analysis based on 16S rRNA gene sequences showed that the strains formed distinct lineages within the genus Hymenobacter. Strains BT523T, BT559T, and 15J16-1T3BT exhibited highest sequence similarities to Hymenobacter armeniacus BT189T (97.6%), Hymenobacter polaris RP-2-7&#xa0;T (97.8%), and Hymenobacter paludis KBP-30&#xa0;T (98.3%), respectively, while strains BT730T, DG25AT, and DG25BT were most closely related to Hymenobacter tibetensis XTM003T, with similarities of 96.3-96.6%. All strains were Gram-negative, aerobic, rod-shaped, and formed red to pink pigmented colonies. Whole-genome analysis revealed genome sizes ranging from 3.78 to 6.33&#xa0;Mb with G&#x2009;+&#x2009;C contents of 55.5-65.0%. Average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between the strains and their closest relatives were below the accepted thresholds for species delineation, supporting their classification as novel species. Functional annotation indicated the presence of genes associated with core metabolism, stress response, and pigment biosynthesis, reflecting adaptation to soil environments. Secondary metabolite analysis further revealed the presence of biosynthetic gene clusters, including terpene and siderophore pathways. Based on polyphasic taxonomic evidence, the six strains are proposed to represent six novel species of the genus Hymenobacter, for which the names Hymenobacter miniatus sp. nov., Hymenobacter madidus sp. nov., Hymenobacter convexus sp. nov., Hymenobacter rubellus sp. nov., Hymenobacter erythromyxa sp. nov., and Hymenobacter radioresistens sp. nov. are proposed. The type strains are BT523T (=&#x2009;KCTC 72341&#xa0;T&#x2009;=&#x2009;NBRC 114851&#xa0;T), BT559T (=&#x2009;KACC 21821&#xa0;T&#x2009;=&#x2009;NBRC 114852&#xa0;T), 15J16-1T3BT (=&#x2009;KCTC 42995&#xa0;T&#x2009;=&#x2009;NBRC 112818&#xa0;T), BT730T (=&#x2009;KACC 22457&#xa0;T&#x2009;=&#x2009;NBRC 116482&#xa0;T), DG25AT (=&#x2009;KCTC 32451&#xa0;T&#x2009;=&#x2009;TBRC 19856&#xa0;T), and DG25BT (=&#x2009;KCTC 32452&#xa0;T&#x2009;=&#x2009;JCM 19444&#xa0;T).

Soil Microbiology↗

Isolation and characterization of two novel species Neorhizobium fuzhouense sp. nov. and Neotabrizicola paludis sp. nov.

Two novel aerobic bacterial strains, designated SGZ-38T and sgz301269T, were isolated from the root of Pennisetum sp. and paddy soil, respectively. Strain SGZ-38T grew at 10-40&#xa0;&#x2103; (optimum 30&#xa0;&#xb0;C) and pH 5.0-12.0 (optimum 6.5) and tolerated up to 1.0% NaCl (w/v), whereas strain sgz301269T grew at 15-37&#xa0;&#xb0;C (optimum 30&#xa0;&#xb0;C), pH 5.0-9.5 (optimum 7.0) and 0-2% NaCl (optimum 0%). Phylogenetic trees based on the 16S rRNA gene and genomes placed both strains into distinct lineages, forming separated clades from their closest relatives. Strain SGZ-381T exhibited the highest 16S rRNA gene similarities to "Neorhizobium deserti" ACCC 61627T (97.4%), and strain sgz301269T had the highest 16S rRNA gene sequence similarity to Neotabrizicola shimadae N10T (97.6%). The respiratory quinone in both strains was ubiquinone-10. The main fatty acids of SGZ-381T were Summed feature 8, Summed feature 2 and C16:0, whereas strain sgz301269T included C10:0 3OH, C18:0 3OH and Summed feature 8. The DNA G+C content of SGZ-381T and sgz301269T was 62.1% and 65.5%, respectively. The average nucleotide identity and digital DNA-DNA hybridization values between each strain and their respective closest species were 74.6% and 20.1%, 75.3% and 17.4% respectively, below the thresholds for species delineation. Based on the comprehensive chemotaxonomic, phylogenetic, and phenotypic evidence, proposed names of the novel strains are Neorhizobium fuzhouense sp. nov. (type strain SGZ-381T=GDMCC1.4207T=JCM 36770T), Neotabrizicola paludis sp. nov. (type strain sgz301269T=MCCC 1K09178T=KCTC 8856T).

Bacterial Typing Techniques↗

Mitsuokella apri sp. nov., a novel species of the family Selenomonadaceae isolated from faeces of a wild boar (Sus scrofa).

A Gram-stain-negative, rod-shaped, obligately anaerobic strain, designated WILCCON 0060T, was isolated from the faecal sample of a wild boar (Sus scrofa)&#xa0;collected on the Island of Ubin (Pulau Ubin), Singapore (1.4126&#xb0;N, 103.9577&#xb0;E). Phylogenetic and comparative analyses based on 16S rRNA gene and whole-genome sequences indicated that strain WILCCON 0060T belongs to the genus Mitsuokella, within the family Selenomonadaceae. Notably, the average nucleotide identity and digital DNA-DNA hybridization values between strain WILCCON 0060T and the type strains of two existing Mitsuokella species, Mitsuokella multacida DSM 20544T and Mitsuokella jalaludinii DSM 13811T, only ranged from 84.6-84.9% and 24.1-24.7%, respectively, both well below the recommended species delineation thresholds of 95.0% and 70.0%. Supported by additional physiological, biochemical and genomic differences, these findings collectively indicate that strain WILCCON 0060T represents a novel species within the genus Mitsuokella. We propose the species name Mitsuokella apri sp. nov., pertaining the isolation of the type strain WILCCON 0060T (=&#x2009;DSM 120052T&#x2009;=&#x2009;LMG 33761T) from a faecal sample of a wild boar.

Animals↗

Genomic and Molecular Interaction Analysis of NodD1 in a Novel Bradyrhizobium yuanmingense sp. B64 Isolate for Nodulation and Symbiosis of Legume Plants.

Rhizobial bacteria are known for their ability to fix nitrogen for leguminous plants and their essential function for sustainable agriculture. This study characterizes the taxonomic status and functional potential of the Bradyrhizobium B64 isolate using integrated genomic and molecular approaches. The whole genome of the B64 isolate was sequenced via Illumina paired-end technology. Species delimitation was performed using average nucleotide identity (ANI) and digital DNA-DNA Hybridization (dDDH). The NodD1 protein structure was modeled using AlphaFold3 and validated by Ramachandran plot analysis. Molecular docking was then conducted to evaluate interactions between NodD1 and four signaling flavonoids: Apigenin, Daidzein, Genistein, and Naringenin. Genomic analysis revealed a maximum ANI of 94.4% and dDDH values between 51.4 and 62.4%. Since these values fall below the standard prokaryotic thresholds (ANI&#x2009;<&#x2009;95%; dDDH&#x2009;<&#x2009;70%), the B64 isolate is identified as a novel species. Physiological assays confirmed nitrogen fixation (1.97 ppm), IAA production (3.67 ppm), and phosphate solubilization (26.10 ppm). Structural validation showed 100% of NodD1 residues in allowed regions, ensuring high model reliability. Docking simulations demonstrated strong binding affinities across all flavonoids, with binding free energies ranging from -&#x2009;8.8 to -&#x2009;9.0&#xa0;kcal/mol. Daidzein exhibited the highest thermodynamic stability (-&#x2009;9.0&#xa0;kcal/mol), whereas apigenin showed the most extensive residue interaction network. The B64 isolate is a novel Bradyrhizobium species with a high symbiotic capacity. The stable NodD1-flavonoid interactions provide a molecular basis for efficient nodulation, positioning B64 as a promising candidate for developing lipo-chitooligosaccharide (LCO)-based biofertilizers.

Bradyrhizobium↗

Genomic Insights Into the Probiotic and Safety Attributes of Pediococcus acidilactici BC-7 for its Potential Application in Livestock Health.

Pediococcus acidilactici is widely recognized for its health-beneficial aspects and has gained increasing interest for use in livestock industry. It shows strong probiotic efficacy, antimicrobial activity, cholesterol-lowering potential, immune modulation, and other therapeutic attributes. The novel strain from indigenous habitats mainly depicted potent probiotic efficacy and high adaptability. In this study, we evaluated the probiotic characteristics and genomic features of strain BC-7 obtained from a Nili - Ravi buffalo calf raised under domestic conditions using phenotypic assessment, genomic analysis and in vivo studies. The strain BC-7 exhibited key probiotic traits i.e., gut tolerance (70.43% - 97.5%), auto-aggregation (85.24%), co-aggregation (14.33% - 25.88%), hydrophobicity (78.33% - 88%), antioxidant potential (54%), and antibacterial activity (16.47-18&#xa0;mm). The safety analysis revealed that BC-7 exhibited susceptibility and resistance to various antimicrobial agents and showed no &#x3b2; hemolytic activity. BC-7 was taxonomically classified as Pediococcus acidilactici by 16&#xa0;S rRNA gene sequencing. Whole genome sequence (WGS) analysis showed that P. acidilactici BC-7 contains a 1.9&#xa0;Mb genome with 42% GC content. Pediococccus acidilactici BC-7 harbored 1900 genes, which were mainly associated with metabolism and genetic processes. Based on genomic comparison, BC-7 shared 99% average nucleotide identity and strong genomic collinearity with P. acidilactici NARCC1 which is a TYPE strain having potent probiotic potential. Probiotic strain BC-7 shared 1,664 core genes with reference strains and 69 unique genes specific for metabolism and genetic processes. The BC-7 strain contained unique bacteriocins-associated genes and defense-related CAzymes, it harbors only vancomycin resistance genes and lacked true virulence determinants. In vivo trial showed that BC-7 treated mice showed increased growth rate, improved immune modulation, and membrane integrity. These significant findings revealed that BC-7 emerging as a potential probiotic strain with strong functionality and efficacy. Thus, our strain BC-7 could be used as a promising candidate for applications in the animal health industry.

Gastrointestinal tract↗

Comparative genomic analysis of Streptococcus parasuis and Streptococcus suis reveals mobile element-associated enrichment of antimicrobial resistance and lack of detectable same-MGE colocalization with virulence-associated genes within stable species boundaries.

Streptococcus suis is a major porcine pathogen and a zoonotic agent that causes meningitis and septicemia in humans. Streptococcus parasuis, a recently recognized close relative, remains poorly characterized with regard to its clinical significance and genomic features. In this study, we generated a single-contig closed genome assembly with genome-wide DNA methylation profiles for S. parasuis strain A1, isolated from a diseased pig in Xinjiang, China, and complemented in silico genomic predictions with isolate-level experimental validation of antimicrobial resistance (AMR) genotypes, virulence genotypes, and phenotypic susceptibility for this reference strain. Using this high-quality genome as a reference anchor, we performed comparative genomic analyses across 195 streptococcal genomes, comprising 15 S. parasuis and 180 S. suis strains, to distinguish genome-level co-occurrence of resistance and virulence determinants from their physical colocalization on the same mobile genetic element (MGE).Species boundaries remained clearly delineated at the genomic level, with a median interspecies average nucleotide identity (ANI) of approximately 86.0%, compared with intraspecies ANI medians of 97.5% for S. parasuis and 96.2% for S. suis. Pangenome analysis identified 12,693 gene clusters, of which 1086 were core clusters, and functional annotation revealed significant differences in accessory gene repertoires between the two species. Within this stable genomic framework, S. parasuis genomes carried a higher AMR gene burden; strain A1 harbored 10 AMR genes, multiple virulence-associated genes, three genomic islands, and eight prophage regions. For strain A1, PCR validation confirmed six AMR genes and six virulence genes, and disk diffusion testing demonstrated a multidrug-resistant phenotype consistent with the genotypic profile.Among 235 predicted mobile elements, 19 harbored AMR genes and seven carried Virulence Factor Database (VFDB) homologs, but none carried both categories simultaneously. This finding reflects a lack of detectable same-MGE colocalization under the applied annotation and assembly framework; it should not be interpreted as evidence of biological physical decoupling. Under a random-placement model, the expected number of co-carrying regions was only 0.57, and the probability of observing zero co-carrying regions was P&#x202f;=&#x202f;0.55. This negative result should be interpreted with caution, given the limited number of cargo-bearing regions and the predominantly draft status of most genomes. Furthermore, the A1 genome contained multiple restriction-modification systems, showed depletion of several methylation motif families in mobile regions, and had limited CRISPR spacer matching evidence, suggesting prior exposure to the relevant sequence space. None of the genomes met our predefined criteria for whole-genome convergence.Collectively, our results support a model in which S. parasuis accumulates AMR-related genes in a modular fashion via mobile elements within stable species boundaries, with no detectable same-MGE colocalization of AMR and virulence determinants under our analytical pipeline. These findings imply that AMR surveillance strategies for this species should prioritize tracking mobile genetic elements rather than inferring wholesale genomic convergence toward S. suis.

Streptococcus suis↗

Tn125-borne blaNDM-1 is decoupled from clonal background in a transcontinental Acinetobacter baumannii ST126/KL14 lineage.

BACKGROUND/OBJECTIVES: Carbapenem-resistant A. baumannii (CRAB) is a WHO Critical Priority pathogen. The blaNDM-1-carrying ST126/KL14 lineage has been independently reported from Vietnam (2015), Malaysia (2016), the USA (2023-2026), and Costa Rica (2024). Whether these geographically distinct reports represent a single transcontinental clone and through what mechanism blaNDM-1 disseminates has not been formally tested. METHODS: We performed comprehensive whole-genome reanalysis of the Vietnamese sentinel isolate DMS06669_L1 using three nested panels (n&#x2009;=&#x2009;19, n&#x2009;=&#x2009;138, and n&#x2009;=&#x2009;609 Vietnamese A. baumannii genomes) and surveyed 429 plasmids extracted from 99 NDM-1 A. baumannii genomes retrieved from NCBI Pathogen Detection. RESULTS: Four ST126/KL14 isolates share high inter-regional average nucleotide identity (ANI; 99.77-99.92%) but wide intra-clade core-SNP distances (41-731 SNPs, well above the &#x223c;20-40-SNP range typical of single-outbreak transmission clusters) and lack a significant molecular clock, consistent with a related transcontinental lineage rather than a single recent clone. NDM-1 plasmid evolution is statistically uncorrelated with chromosomal sequence type (Spearman &#x3c1;&#x2009;=&#x2009;0.131, P&#x2009;=&#x2009;0.573). At 609-strain population scale, under a fragmentation-aware detection criterion, all 41 blaNDM-1-carrying Vietnamese strains also carry ISAba125, with none carrying blaNDM-1 without it (Fisher exact test; Haldane-Anscombe-corrected OR&#x2009;&#x2248;2.0 &#xd7; 10&#xb3;, 95% CI 1.2 &#xd7; 10&#xb2; to 3.4 &#xd7; 10&#x2074;; P&#x2009;=&#x2009;4.74&#xd7;10&#x207b;&#x2074;&#x2078;; &#x3c6;&#x2009;=&#x2009;0.79). CONCLUSIONS: The blaNDM-1 dissemination pattern in this ST126/KL14 lineage is primarily consistent with Tn125 transposition acting alongside plasmid-borne spread. Standard MLST-based surveillance is insufficient; multi-level genomic monitoring - including chromosomal and plasmid-level detection of the Tn125/ISAba125 unit - is required to track this resistance threat.

A. baumannii↗

Genomic characterization of a hypervirulent Aeromonas veronii NN0115 from Nile tilapia and head kidney transcriptome of infected fish reveals B-cell-dominated immune response with specific immunoglobulin downregulation.

Aeromonas veronii is a pathogen of multiple fish species, yet systematic understanding of its infection in Nile tilapia (Oreochromis niloticus) remains limited. A dominant strain, NN0115, was isolated from a natural outbreak and identified as A. veronii by 16S rRNA and whole-genome average nucleotide identity (ANI, 96.33%). Experimental infection revealed high virulence (LD50&#x202f;=&#x202f;3.41&#x202f;&#xd7;&#x202f;106&#x202f;CFU/mL, equivalent to 8.53&#x202f;&#xd7;&#x202f;104&#x202f;CFU/fish). The genome is 4.58&#x202f;Mb (58.57% GC) and encodes 4216 proteins. Virulence factor analysis identified 1253 genes, dominated by motility-related (264) and immune modulation (208) factors. Genomic island GI2 harbors 7 virulence genes and two dual-function resistance-virulence genes. The strain is resistant to 9 of 25 agents tested but carries three RND efflux pump genes whose predicted resistance was not phenotypically observed. The head kidney transcriptome of tilapia at 24&#x202f;h post-bacterial infection identified 773 differentially expressed genes; among them, 57 were immunoglobulin (Ig) genes, and 56 were down-regulated. Integration of published single-cell transcriptomic data showed that non-Ig B-cell marker genes were down-regulated by 32%, whereas Ig genes were reduced by 63%, indicating selective transcriptional suppression of Ig genes rather than a general decrease in B-cell transcriptional activity. Together, this study provides a comprehensive characterization of a highly virulent A. veronii from Nile tilapia and reveals that selective downregulation of B-cell Ig genes is the dominant transcriptional feature of the host head kidney response.

Animals↗

Gloeotrichia echinulata genomes from the United States are nontoxigenic and likely geosmin producers.

Six Gloeotrichia echinulata genomes derived from planktonic harmful algal blooms (HABs) with similar colonial morphology have been sequenced from lakes in the west and northeast regions of USA, four of them to completion. The c. 7 Mbp genomes exhibit a high level of conservation, with 98-99% pairwise genome-wide average nucleotide identity and high levels of synteny, representing a single species cluster. We observed strong conservation of gene clusters responsible for the synthesis of the secondary metabolites and bioactive peptides that are characteristic of HAB-forming cyanobacteria. All six G. echinulata genomes lack genes for the synthesis of classic cyanotoxins, including microcystin, but possess genes responsible for the synthesis of the taste and odor compound geosmin. Interestingly, the geoA geosmin synthase gene in three genomes is homologous to other cyanobacterial geoA genes, while the other three geoA genes are related to actinomyces geoA. Phylogenomic analysis places the G. echinulata genomes within a clade of benthic Nostocales, reflecting an ecological niche featuring extensive growth on the sediment surface before colonies disperse into the epilimnion for planktonic growth. We identify genes conserved in all six genomes that could represent physiological adaptations supporting active growth on sediments and pelagic recruitment independent of wind-driven mixing: phycoerythrin light harvesting complexes for optimal photosynthesis at depth; gliding motility to access patchy nutrient distributions; and gas vesicles with relatively small GvpC proteins that predict resistance to higher hydrostatic pressure. The strong genomic similarity across geographically distant populations suggests that G. echinulata in the United States is a tightly related non-toxigenic species group with predictable properties relevant to public health and drinking water management.

Cyanobacteria↗

Characterisation of Carbapenem-Resistant Raoultella planticola and Structural Analysis of NDM Composite Plasmids.

OBJECTIVE: This study aimed to investigate the molecular characteristics, resistant plasmid structures and phylogeny of a carbapenem-resistant Raoultella planticola (CRRP) strain from a patient with pneumonia to inform antimicrobial resistance control strategies. METHODS: We performed strain identification using MALDI-TOF MS, the BD Phoenix 100 system and whole-genome sequencing (WGS). We assessed antimicrobial susceptibility and resistance gene transfer using PCR, conjugation and stability assays, plasmid structure using a bioinformatics tool and phylogeny using a core-genome phylogenetic tree. RESULTS: WGS confirmed the isolate as R. planticola (average nucleotide identity (ANI) > 98.9% with reference type strains), co-harbouring blaKPC-2 and blaNDM-1. It was resistant to 19 antimicrobial agents and susceptible to only polymyxin, amikacin and chloramphenicol. Resistance genes were present on two conjugative plasmids: pzwx_KPC (IncFIA) and pzwx_NDM (a novel repFIB/repHI5B hybrid assembled via non-homologous end joining). Both plasmids demonstrated efficient transfer and stable inheritance over 12 passages. pzwx_KPC was highly homologous to plasmids from Klebsiella pneumoniae. Phylogenetic analysis revealed the closest relationship with German R. planticola strains. CONCLUSION: CRRP carries highly transmissible and stable resistance plasmids. Strengthened monitoring in immunocompromised patients and improved environmental disinfection are recommended. The risk of misidentification by automated systems underscores the importance of WGS for accurate pathogen identification.

Carbapenem resistance↗