Bacteriophages HungryHenry, CaptainRex, and ChikPic were isolated from soil collected in Tennessee using the bacterium Microbacterium foliorum . These bacteriophages have genomes that are 41,516 bp, 39,941 bp, and 40,333 bp in length and encode 62, 61, and 63 putative genes, respectively. Based on gene content similarity, all three bacteriophages are assigned to actinobacteriophage cluster EA (subclusters EA1, EA2 and EA5, respectively).

This announcement reports the complete genome sequences of two bacteriophages isolated from soil samples using the host Arthrobacter atrocyaneus Strain NRRL B-2883. These findings enhance our understanding of AZ1 cluster phages, particularly Wildwest and Sue2, with their unique genomic features.

Phage therapy, which uses phages to decrease bacterial load in an ecosystem, introduces a multitude of gene copies (bacterial and phage) into said ecosystem. While it is widely accepted that phages have a significant impact on ecology, the mechanisms underlying their impact are not well understood. It is therefore paramount to understand what is released in the said ecosystem, to avoid alterations with difficult-to-predict-but potentially huge-consequences. An in-depth annotation of therapeutic phage genomes is therefore essential. Currently, the average published phage genome has only 20-30% functionally annotated genes, which represents a hurdle to overcome to deliver safe phage therapy, for both patients and the environment. This study aims to compare the effectiveness of manual versus automated phage genome annotation methods. Twenty-seven phage genomes were annotated using SEA-PHAGE and Rime Bioinformatics protocols. The structural (gene calling) and functional annotation results were compared. The results suggest that during the structural annotation step, the SEA-PHAGE method was able to identify an average of 1.5 more genes per phage (typically a frameshift gene) and 5.3 gene start sites per phage. Despite this difference, the impact on functional annotation appeared to be limited: on average, 1.2 genes per phage had erroneous functions, caused by the structural annotation. Rime Bioinformatics' tool (rTOOLS, v2) performed better at assigning functions, especially where the SEA-PHAGE methods assigned hypothetical proteins: 7.0 genes per phage had a better functional annotation on average, compared to SEA PHAGE's 1.7. The method comparison detailed in this article indicate that (1) manual structural annotation is marginally superior to rTOOLS automated structural annotation; (2) rTOOLS automated functional annotation is superior to manual functional annotation. Previously, the only way to obtain a high-quality annotation was by using manual protocols, such as SEA-PHAGES. In the relatively new field of phage therapy, which requires support to advance, manual work can be problematic due to its high cost. Rime Bioinformatics' rTOOLS software allows for time and money to be saved by providing high-quality genome annotations that are comparable to manual results, enabling a safer and faster-developing phage therapy.

We report the discovery and characterization of bacteriophages ASegato, DejaVu, Judebell, and RicoCaldo, isolated from grass samples collected in Quincy, Massachusetts, using Microbacterium foliorum B-24224 as the isolation host. Based on gene content similarity, these phages are assigned to actinobacteriophage clusters ED2, ED1, EG, and EK2 respectively.

Among enduringly associated parasitic taxa, tapeworms (Platyhelminthes: Cestoda), particularly the family Anthocephaliidae Ruhnke, Caira & Cox, 2015, pose challenges to systematic history of the group. Within this family, the genus Anthocephalum Linton, 1890, remains insufficiently explored despite recent advancements. This study addresses the intricate taxonomy, phylogenetic relationships, diversity, and historical biogeography of tapeworms within the Anthocephaliidae, focusing on its type-genus Anthocephalum. To accomplish this objective, 15 specimens across various geographical regions including Alagoas and Pará in Brazil, Panama, and Senegal were selected for DNA extraction. For species of Anthocephalum, partial 18S and 28S rDNA sequences were amplified via PCR, purified, and sequenced using the ABI Big-Dye method. Three specimens of Alveobothrium grabatum Boudaya, Neifar & Euzet, 2018 were sequenced through Illumina technology. Phylogenetic analyses were conducted using IQTREE2 under two optimality criteria: Maximum Parsimony (MP) and Maximum Likelihood (ML). Biogeographic ancestral range estimations were performed using the R package BioGeoBEARS, incorporating multiple biogeographical models. Our phylogenetic analyses reaffirmed Anthocephaliidae's monophyly. However, both the relationships with or within Anthocephaliidae require further investigation. One example within the family is the positioning of Alveobothrium Boudaya, Neifar & Euzet, 2018, which challenged Anthocephalum's monophyly requiring taxonomic actions. Furthermore, when exploring new localities and/or new hosts for Anthocephalum, four independent lineages were identified, suggesting that Anthocephalum's diversity remains underestimated especially in unexplored regions and hosts. About the biogeographic ancestral range estimation, the analysis suggested a Central Indo-Pacific origin for early Anthocephalum lineages, with subsequent colonization events shaping the current diversity. The preliminary biogeographical framework presented here underscores the importance of refining phylogenetic hypotheses and enhancing taxonomic understanding. As taxonomic actions taken, the four new lineages were formally described and the genus Alveobothrium was synonymized with Anthocephalum, for which we proposed an amended diagnosis. This revision brings the total number of valid species of Anthocephalum to 30. Therefore, we suggest that future research initiatives should prioritize time-calibrated analyses, multiple genetic loci, and broader taxonomic representation for a detailed exploration of systematic history of anthocephaliid's.

The rise of herbicide-resistant weeds like Conyza canadensis L. poses a challenge to modern agriculture, driving the need for eco-friendly alternatives. Microbial metabolites from actinobacteria species offer promising weed-control solutions. This study aims to screen and identify an actinobacteria isolate from Brazil's Caatinga biome that produces phytotoxic metabolites and to characterize its compounds.

An isolate, named as Caat 7-52, was selected because of its significant phytotoxic effects against Lemna minor L. Phylogenetic analyses using six concatenated genes (gyrB, recA, rpoB, trpB, atpD and 16S rRNA) confirmed Caat 7-52's close relationship to Streptomyces musisoli TBRC 9950T, despite phenotypical differences. Bioassay-directed isolation against L. minor revealed 3-hydroxybenzoic acid and albocycline as phytotoxins, with minimum inhibitory concentrations of 50.00 and 3.12 μg mL-1, respectively. Albocycline analogues were also detected and exhibited moderate phytotoxicity in L. minor. In addition, albocycline effectively inhibited the seed germination of C. canadensis with a minimum inhibitory concentration of 6.25 μg mL-1, marking the first report of albocycline's phytotoxic activity. Direct use of the fermented broth selectively inhibited dicot weeds, offering a sustainable and solvent-free weed management strategy.

The discovery of Streptomyces sp. Caat 7-52 and its metabolites, combined with the direct application of fermented broth, represents a significant advancement in sustainable weed control. This bioherbicidal approach offers an environmentally friendly alternative for managing resistant weeds like C. canadensis and supports the broader use of microbial metabolites in integrated pest management programs. © 2025 Society of Chemical Industry.

AWGoat, a lytic bacteriophage that infects Arthrobacter globiformis, was isolated from a goat pen in Charlotte, NC. Its genome is 65,496 bp long, with a GC content of 67.1%. Based on gene content similarity, AWGoat is assigned to actinobacteriophage cluster AP. It encodes a putative toxin from a broadly distributed toxin/antitoxin pair and an unusually large minor tail protein.

Bacteriophages Biscayne, Bush and GreenIvy were isolated from soil samples in Miami, FL using Microbacterium foliorum NRRL B-24224 as host. Transmission electron microscopy shows siphoviral morphologies for all three phages. Based on gene content similarity to other actinobacteriophages, they are assigned to the EE, GA and EA5 clusters, respectively.

Bacteriophage WestPM is a siphoviral-like phage infecting Microbacterium foliorum isolated from environmental samples collected on Pensacola Beach, FL. The genome of this phage is 39,693 bp long and contains 59 predicted protein-coding genes and zero tRNA genes. Based on gene content similarity, WestPM is grouped in the actinobacteriophage EA11 subcluster.

Invasive alien species have the potential to introduce pathogens of economic and health importance in new environments. In Brazil, parasites from the non-native European brown hare can be a threat to humans, domestic animals, and wildlife. Therefore, we aimed to describe the helminth fauna of the invasive European brown hare in three Brazilian states (São Paulo, Paraná, and Rio Grande do Sul). For this, 90 brown hares were collected and examined for helminths. Helminth specimens recovered were morphologically identified and genetically characterized based on the DNA of male specimens using three genetic regions (28S rDNA, ITS-2, and cox-1 mtDNA). Descriptors of infection were calculated, and statistical analysis was performed. Parasites were found only in the small intestine of 14.4% (13/90) of brown hares and low parasite loads per animal were recorded (range = 1-530). The obtained specimens were morphologically identified as Trichostrongylus colubriformis and Bunostomum trigonocephalum. There was a high level of agreement between phylogenetic analysis and morphology for T. colubriformis. The geographical region was the only significant factor for infection; the State of Rio Grande do Sul had a higher general prevalence, higher T. colubriformis specific prevalence, and higher mean abundance than the other states evaluated. All hares were in a good body condition. To our knowledge, this is a new host record for B. trigonocephalum. This is the first study on the helminthological diversity of European brown hares in Brazil, and our results indicate that their helminth fauna is represented by parasites of domestic ruminants with zoonotic potential.

Uptake hydrogenase (Hup) recycles H2 formed by nitrogenase during nitrogen fixation, thereby preserving energy. Among root nodule bacteria, most rhizobial strains examined are Hup-, while only one Hup-  Frankia inoculum had been identified. Previous analyses had led to the identification of two different [NiFe] hydrogenase syntons. We analysed the distribution of different types of [NiFe] hydrogenase in the genomes of different Frankia species. Our results show that Frankia strains can contain four different [NiFe] hydrogenase syntons representing groups 1f, 1h, 2a, and 3b according to Søndergaard, Pedersen, and Greening (HydDB: a web tool for hydrogenase classification and analysis. Sci Rep 2016;6:34212. https://doi.org/10.1038/srep34212.); no more than three types were found in any individual genome. The phylogeny of the structural proteins of groups 1f, 1h, and 2a follows Frankia phylogeny; the phylogeny of the accessory proteins does not consistently. An analysis of different [NiFe] hydrogenase types in Actinomycetia shows that under the most parsimonious assumption, all four types were present in the ancestral Frankia strain. Based on Hup activities analysed and the losses of syntons in different lineages of genome reduction, we can conclude that groups 1f and 2a are involved in recycling H2 formed by nitrogenase while group 1 h and group 3b are not.

Mycobacteriophages FireRed, MISSy, MPhalcon, Murica, Sassay, Terminus, Willez, YassJohnny, and Youngblood were isolated from soil using Mycobacterium smegmatis as a host. Genome sequencing and annotation revealed that they belong to Actinobacteriophage Cluster E. Here, we describe the features of their genomes and discuss similarities within these Cluster E phages.

Bacteriophage GiJojo is a myovirus isolated from soil that infects Streptomyces mirabilis NRRL B-2400, with a genome length of 115,161 bp containing 180 genes and 29 tRNAs. Of those genes, 59 have been assigned functions. GiJojo is a member of the BS cluster of actinobacteriophages.

Xenia2 is a DV cluster actinobacteriophage that infects Gordonia rubripertincta NRRL B-16540. The genome is 68,135bp, has a GC content of 57.9% and 98 predicted protein-coding genes, 33 of which have a predicted function. Xenia2 has a lysis cassette with an endolysin (lysin A) and four different holin-like transmembrane proteins.

We present the bacteriophages GoblinVoyage and Doxi13, siphoviruses isolated on Streptomyces scabiei RL-34. They belong to the BI2 cluster and have genomes consisting of 60.9% GC content with identical 3' end sticky overhangs. The genome lengths of GoblinVoyage and Doxi13 are 43,540 bp and 43,696 bp, respectively.

Bacteriophage Curie is a podovirus that infects Microbacterium foliorum. The Curie genome spans 16,810 bp, has 90 bp terminal inverted repeats, and includes 23 protein-coding genes. Its genome architecture resembles phage PineapplePizza and other phi29-like phages. Together, Curie and PineapplePizza form a new actinobacteriophage Cluster GI.

Senecavirus A (SVA) is a picornavirus that is endemic in swine, causing a vesicular disease clinically indistinguishable from other vesicular diseases, like foot-and-mouth disease. The widespread viral circulation, constant evolution, and economic losses caused to the swine industry emphasize the need for measures to control the agent. In this study, we evaluated the immunogenicity of a whole-virus-inactivated vaccine using a representative contemporary Brazilian SVA strain in Balb/ByJ mice. The animals were vaccinated with two doses by an intramuscular route. The humoral response induced by the vaccination was evaluated by an in-house ELISA assay for IgG detection. The cellular response was assessed by flow cytometry after in vitro SVA stimulation in splenocyte cultures from vaccinated and non-vaccinated groups. Protection against SVA was assessed in the experimental groups following an oral challenge with the homologous virus. The vaccination induced high levels of IgG antibodies and the proliferation of CD45R/B220+sIgM+, CD3e+CD69+, and CD3e+CD4+CD44+CD62L- cells. These results indicate the immunogenicity and safety of the vaccine formulation in a murine model and the induction of humoral and cellular response against SVA.

Four rod-shaped, non-motile, non-spore-forming, facultative anaerobic, Gram-stain-positive lactic acid bacteria, designated as EB0058T, SCR0080, LD0937T and SCR0063T, were isolated from different corn and grass silage samples. The isolated strains were characterized using a polyphasic approach and EB0058T and SCR0080 were identified as Lacticaseibacillus zeae by 16S rRNA gene sequence analysis. Based on whole-genome sequence-based characterization, EB0058T and SCR0080 were separated into a distinct clade from Lacticaseibacillus zeae DSM 20178T, together with CECT9104 and UD2202, whose genomic sequences are available from NCBI GenBank. The average nucleotide identity (ANI) values within the new subgroup are 99.9 % and the digital DNA-DNA hybridization (dDDH) values are 99.3-99.9 %, respectively. In contrast, comparison of the new subgroup with publicly available genomic sequences of L. zeae strains, including the type strain DSM 20178T, revealed dDDH values of 70.2-72.5 % and ANI values of 96.2-96.6 %. Based on their chemotaxonomic, phenotypic and phylogenetic characteristics, EB0058T and SCR0080 represent a new subspecies of L. zeae. The name Lacticaseibacillus zeae subsp. silagei subsp. nov. is proposed with the type strain EB0058T (=DSM 116376T=NCIMB 15474T). According to the results of 16S rRNA gene sequencing, LD0937T and SCR0063T are members of the Lacticaseibacillus group. The dDDH value between the isolates LD0937T and SCR0063T was 67.6 %, which is below the species threshold of 70 %, clearly showing that these two isolates belong to different species. For both strains, whole genome-sequencing revealed that the closest relatives within the Lacticaseibacillus group were Lacticaseibacillus huelsenbergensis DSM 115425 (dDDH 66.5 and 65.9 %) and Lacticaseibacillus casei DSM 20011T (dDDH 64.1 and 64.9 %). Based on the genomic, chemotaxonomic and morphological data obtained in this study, two novel species, Lacticaseibacillus parahuelsenbergensis sp. nov. and Lacticaseibacillus styriensis sp. nov. are proposed and the type strains are LD0937T (=DSM 116105T=NCIMB 15471T) and SCR0063T (=DSM 116297T=NCIMB 15473T), respectively.

Amabiko is a lytic subcluster BE2 bacteriophage that infects Streptomyces scabiei-a bacterium causing common scab in potatoes. Its 131,414 bp genome has a GC content of 49.5% and contains 245 putative protein-coding genes, 45 tRNAs, and one tmRNA. Amabiko is closely related to Streptomyces bacteriophage MindFlayer (gene content similarity: 86.5%).

Here, we present bacteriophage SoJo, a siphovirus infecting Streptomyces mirabilis, with a circularly permuted genome of 39 kbp and GC content of 71.5%. Its genome length and content are similar to that of other phages in the Actinobacteriophage Database BC cluster. SoJo was isolated from soil in Columbia, MD, USA.

The Gram-positive bacteria Streptomyces davaonensis and Streptomyces cinnabarinus have been the only organisms known to produce roseoflavin, a riboflavin (vitamin B2) derived red antibiotic. Using a selective growth medium and a phenotypic screening, we were able to isolate a novel roseoflavin producer from a German soil sample. The isolation procedure was repeated twice, that is, the same strain could be isolated from the same location in Berlin 6 months and 12 months after its first isolation. Whole genome sequencing of the novel roseoflavin producer revealed an unusual chromosomal arrangement and the deposited genome sequence of the new isolate (G + C content of 71.47%) contains 897 genes per inverted terminal repeat, 6190 genes in the core and 107 genes located on an illegitimate terminal end. We identified the roseoflavin biosynthetic genes rosA, rosB and rosC and an unusually high number of riboflavin biosynthetic genes. Overexpression of rosA, rosB and rosC in Escherichia coli and enzyme assays confirmed their predicted functions in roseoflavin biosynthesis. A full taxonomic analysis revealed that the isolate represents a previously unknown Streptomyces species and we propose the name Streptomyces berlinensis sp. nov. for this roseoflavin producer.

Belonging to the Actinobacteria phylum, members of the Rhodococcus genus thrive in soil, water, and even intracellularly. While most species are non-pathogenic, several cause respiratory disease in animals and, more rarely, in humans. Over 100 phages that infect Rhodococcus species have been isolated but despite their importance for Rhodococcus ecology and biotechnology applications, little is known regarding the molecular genetic interactions between phage and host during infection. To address this need, we report RNA-Seq analysis of a novel Rhodococcus erythopolis phage, WC1, analyzing both the phage and host transcriptome at various stages throughout the infection process.

By five minutes post-infection WC1 showed upregulation of a CAS-4 family exonuclease, putative immunity repressor, an anti-restriction protein, while the host showed strong upregulation of DNA replication, SOS repair, and ribosomal protein genes. By 30 min post-infection, WC1 DNA synthesis genes were strongly upregulated while the host showed increased expression of transcriptional and translational machinery and downregulation of genes involved in carbon, energy, and lipid metabolism pathways. By 60 min WC1 strongly upregulated structural genes while the host showed a dramatic disruption of metal ion homeostasis. There was significant expression of both host and phage non-coding genes at all time points. While host gene expression declined over the course of infection, our results indicate that phage may exert more selective control, preserving the host's regulatory mechanisms to create an environment conducive for virion production.

The Rhodococcus genus is well recognized for its ability to synthesize valuable compounds, particularly steroids, as well as its capacity to degrade a wide range of harmful environmental pollutants. A detailed understanding of these phage-host interactions and gene expression is not only essential for understanding the ecology of this important genus, but will also facilitate development of phage-mediated strategies for bioremediation as well as biocontrol in industrial processes and biomedical applications. Given the current lack of detailed global gene expression studies on any Rhodococcus species, our study addresses a pressing need to identify tools and genes, such as F6 and rpf, that can enhance the capacity of Rhodococcus species for bioremediation, biosynthesis and pathogen control.

PSonyx is a newly isolated phage that infects Corynebacterium glutamicum. This siphovirus was isolated from a French pond in the south of Paris by students from Paris-Saclay University. Its 80,277-bp singleton genome carries 136 protein-coding genes and 5 tRNAs.

We report the discovery and genome sequence of CandC, a lytic bacteriophage with siphovirus morphology. CandC was isolated from a soil sample from Plattsburgh, NY, USA (Fall 2021). It has a genome size of 62,344 bp with 106 predicted protein-encoding genes, 30 of which are assigned putative functions.

Phage Culver, with a siphovirus morphology, was isolated using Gordonia terrae CAG3. Culver is assigned to phage cluster CQ1 based on gene content similarity to actinobacteriophages. Notably, Culver is predicted to encode eight tRNAs, lysin A by two adjacent genes, and, unlike other CQ1 phages, two putative integrase genes.

Fungi colonizing plants are gaining attention because of their ability to promote plant growth and suppress pathogens. While most studies focus on endosymbionts from grasses and legumes, the large and diverse group of ericaceous plants has been much neglected. We recently described one of the very few fungal endophytes promoting the growth of the Ericaceae Vaccinium macrocarpon (American cranberry), notably the Codinaeella isolate EC4. Here, we show that EC4 also suppresses fungal pathogens, which makes it a promising endophyte for sustainable cranberry cultivation. By dual-culture assays on agar plates, we tested the potential growth suppression (or biocontrol) of EC4 on other microbes, notably 12 pathogenic fungi and one oomycete reported to infect not only cranberry but also blueberry, strawberry, tomato plants, rose bushes and olive trees. Under greenhouse conditions, EC4 protects cranberry plantlets infected with one of the most notorious cranberry-plant pathogens, Diaporthe vaccinii, known to cause upright dieback and berry rot. The nuclear genome sequence of EC4 revealed a large arsenal of genes potentially involved in biocontrol. About ∼60 distinct clusters of genes are homologs of secondary metabolite gene clusters, some of which were shown in other fungi to synthesize nonribosomal peptides and polyketides, but in most cases, the exact compounds these clusters may produce are unknown. The EC4 genome also encodes numerous homologs of hydrolytic enzymes known to degrade fungal cell walls. About half of the nearly 250 distinct glucanases and chitinases are likely involved in biocontrol because they are predicted to be secreted outside the cell. Transcriptome analysis shows that the expression of about a quarter of the predicted secondary-metabolite gene clusters and glucan and chitin-degrading genes of EC4 is stimulated when it is co-cultured with D. vaccinii. Some of the differentially expressed EC4 genes are alternatively spliced exclusively in the presence of the pathogen, altering the proteins' domain content and subcellular localization signal, thus adding a second level of proteome adaptation in response to habitat competition. To our knowledge, this is the first report of Diaporthe-induced alternative splicing of biocontrol genes.

We report the genome sequences of 31 mycobacteriophages isolated on Mycobacterium smegmatis mc2155 at room temperature. The genomes add to the diversity of Clusters A, B, C, G, and K. Collectively, the genomes include 70 novel protein-coding genes that have no close relatives among the actinobacteriophages.