Complete Genome Research Articles

Complete genome sequences of Pantoea vagans strains GM1 and GM2, isolated from the leaves of garlic mustard.

We report the complete genomes of two Pantoea vagans strains, GM1 and GM2, isolated from garlic mustard plants. P. vagans strain GM1 was found to carry a HiVir pantaphos biosynthetic gene cluster, which causes onion necrosis.

Complete genomes of common bacterial hosts of RNA bacteriophage Φ6.

Bacterial plant pathogens Pseudomonas savastanoi pv. phaseolicola, P. syringae pv. tomato, P. syringae pv. atrofaciens, and P. oleovorans East River isolate A serve as common laboratory hosts for bacteriophage Φ6, a widely used model organism for enveloped viruses. Here, we report complete genome sequences for strains of these bacterial hosts.

Complete Genome Sequence and In Vitro Probiotic Assessment of Bacillus subtilis DC-11 Isolated from Traditionally Fermented Idli Batter.

In this study, we reported in vitro probiotic assessment and complete genome sequence of Bacillus subtilis DC-11 isolated from traditionally fermented Idli Batter. The strain was evaluated for probiotic properties, biofilm formation, and antimicrobial compound production. The phenotypic safety was determined by accessing the strain's ability to produce enterotoxins, degrade mucin, and antibiotic sensitivity. Whole genome sequencing (WGS) was performed to identify the strain and determine genetic safety by analyzing the presence of plasmids, antibiotic resistance genes, and virulence factors. In the results, B. subtilis DC-11 showed 88.98% viability in gastric juice, and 98.60% viability in intestinal juice. It showed 18.33 ± 0.44% autoaggregation, 32.53 ± 3.11% adhesion to xylene, 0.98 ± 0.05 OD unit's adhesion to mucin (crystal violet equivalence at 550nm), 21.2 ± 2.3% adhesion to Caco-2 cells, and - 22.3 ± 0.65mV zeta potential. The highest co-aggregation was recorded with Escherichia coli (23.62 ± 0.70%). The strain was found negative for enterotoxin production, mucin degradation, and antibiotic resistance to the commonly used therapeutic antibiotics. It formed a good biofilm and capable of producing antimicrobial peptide subtilosin A with a molecular mass of 3400Da. The peptide has inhibited the growth of methicillin-resistant Staphylococcus aureus (18.6 ± 0.58mm). In genetic safety, no plasmids, antibiotic-resistant genes, and virulence factors were detected. Moreover, the strain showed close similarity with B. subtilis ATCC 6051 and proteins involved in probiotic attributes. In conclusion, B. subtilis DC-11 is safe potential probiotic candidate.

Complete genome sequence of a marine Pseudoalteromonas bacterial strain.

Pseudoalteromonas is an abundant bacterial genera, found ubiquitously, including in extreme environments. Its broad metabolic capacity enables unique associations with various organisms. Using PacBio sequencing, we generated the complete genome sequence of a marine Pseudoalteromonas, revealing two circular chromosomes and one putative plasmid. The genome data are accessible at https://BacBrowse.univ-nantes.fr.

Complete genome sequence of a human influenza a virus (H1N1) detected in Kazakhstan in the fall of 2022.

An Orthomyxoviridae Alphainfluenza virus Influenza A virus strain, A/Kazakhstan/Flu-H11-6/2022(H1N1), was isolated in Karaganda, Central Kazakhstan during a study of acute respiratory infections among hospital inpatients in 2022. Here, we present the complete genome sequence of this strain.

Comparative study of two Rift Valley fever virus field strains originating from Mauritania.

Rift Valley fever (RVF) is one of the major viral arthropod-borne diseases in Africa. In recent decades, RVF virus (RVFV), the causative agent of RVF, has been responsible for multiple outbreaks in West Africa with important consequences on human and animal health. In particular, an outbreak occurred in 2010 after heavy rains in the desertic region of Adrar, Mauritania. It was characterized by the appearance of severe clinical signs among dromedary camels. Another one occurred in 2013-2014 across Senegal and the southern part of Mauritania. In this study, we characterized two RVFV field strains isolated during these two outbreaks. The first strain, MRU25010-30, was isolated from a camel (2010) while the second, MRU2687-3, was isolated from a goat (2013). By deep-sequencing and rapid amplification of cDNA-ends by polymerase chain reaction, we successfully sequenced the complete genome of these two RVFV strains as well as the reference laboratory strain ZH548. Phylogenetic analysis showed that the two field viruses belong to two different RVFV genetic lineages. Moreover, we showed that MRU25010-30 replicates more efficiently in various in vitro cell culture models than MRU2687-3 and ZH548. In vivo, MRU25010-30 caused rapid death of BALB/c mice and proved to be more virulent than MRU2687-3, regardless of the route of inoculation (subcutaneous or intranasal). The virulence of MRU25010-30 is associated with a high viral load in the liver and serum of infected mice, while the death of mice infected with MRU2687-3 and ZH548 correlated with a high viral load in the brain. Altogether, the data presented in this study provide new avenues to unveil the molecular viral determinants that modulate RVFV virulence and replication capacity.

Genetic Diversity of Cotton Leafroll Dwarf Virus from the Southwestern United States and Its Implications for the Multi-Introduction Event Hypothesis and Future Evolution.

Cotton leafroll dwarf virus (CLRDV) is a viral agent recently identified in the United States in 2017 in Alabama. Since its identification, CLRDV has spread to every cotton-growing state east of New Mexico. Oklahoma, Kansas, and Texas make up the westernmost border of reported CLRDV incidence, making monitoring of these states vital for proper control. Additionally, as the virus evolves, mutations that alter symptomology, such as mutations in the F-box-like motif in ORF0/P0, may occur and need to be monitored thoroughly during the growing seasons. Using high-throughput sequencing and PCR-derived Sanger sequencing, 4 CLRDV genomes and 21 P0 gene isolates were sequenced from Oklahoma, Kansas, and Texas from 2019 to 2021 to determine the genetic diversity among CLRDV isolates. Phylogenetic analyses of the complete genomes revealed seven clades, whereas ORF0 gene analyses resulted in large polytomic clusters. BEAST analyses of the 114 total P0 sequences from GenBank, downloaded before 2024, revealed a lower mean substitution rate than previously reported as well as an earlier root year (1914). In addition, using all available CLRDV genome sequences, 11 likely recombination events were determined. Examination of the P0 amino acid sequences revealed 13 mutations unique to the isolates collected in this study. Based on the phylogenetic and amino acid analyses, the CLRDV isolates from Texas (TX clade) may represent evidence for the multi-introduction event hypothesis into the United States. Additionally, based on our analyses in this study, we propose the Asian CLRDV isolates should be constituted as a potentially separate strain of CLRDV.

Characterization of the Mitochondrial Genome of Cambaroides schrenckii (Astacidea: Cambaridae) and Its Phylogenetic Implications

Background: Cambaroides schrenckii is an endangered freshwater crayfish in China, belonging to the genus Cambaroides, that can act as a complementary host for paragonimus. The objective of this study was to examine the complete mitochondrial genome characteristics and their evolutionary relationships within the Astacidea. Methods: The analysis of gene rearrangements and evolutionary relationships was conducted through the sequencing of the mitochondrial genome of C. schrenckii. Results: C. schrenckii mitochondrial genome length was 15,572, comprising thirteen PCGs, two rRNAs, 22 tRNAs, and one d-loop region of C. schrenckii. The mitochondrial genome of C. schrenckii exhibits an A + T content of 69.61% and a G + C content of 30.39%. Among the thirteen PCGs, cytb, nad3, and nad6 have a start codon of ATT, while the other ten PCGs have ATC, ATA, and ATG start codons. All 22 tRNA genes displayed a typical cloverleaf secondary structure. Gene rearrangement analysis showed that seven gene arrangements were identified based on PCGs in the infraorder Astacidea, with type I being the most common. Conclusions: The relationship between the American Cambaridae is closer to Astacidae than the Asian Cambaridae. The present study provides a theoretical basis for further discussions of developmental relationships in the infraorder Astacidea.

Discovery of two novel foamy viruses in sea lions and dolphins provides insight into their evolutionary history.

The prevalence and evolution of foamy viruses (FVs) have become the focus of research because of the risk of new zoonotic diseases. FVs have been isolated from various mammals and exhibit long-term co-speciation with their hosts. They also appear to be mild and nonpathogenic to their hosts. However, they may increase the risk of infection by other pathogens or exacerbate the symptoms of other diseases. Based on the data obtained using next-generation sequencing (NGS), we amplified and obtained the complete genomes of the two new FVs discovered in the bottlenose dolphin (Tursiops truncatus) and the South American sea lion (Otaria byronia) at the Qingdao Polar Haichang Ocean Park. Analysis and prediction of the novel FV's genomic structure revealed that it was consistent with that of the known mammalian FVs. The polmerase (pol) genes of the novel OFVoby_1 and DFVttr_1 showed less than 61.87 % and 61.83 % amino acid identity, respectively, with other known FVs belonging to the Retroviridae family. The host was likely to carry the FV for a considerable amount of time, as evidenced by the different times DFVttr_1 was discovered. The phylogenetic analysis revealed that the pol of OFVoby_1 and DFVttr_1 closely clustered with the FVs of Simiispumavirus and Felispumavirus, respectively. However, they both displayed distinct branches. According to the international committee on taxonomy of viruses (ICTV) FV classification criteria, FVs carried by dolphins and sea lions belong to two new genera within the Spumaretrovirinae subfamily. Using Bayesian analysis to simultaneously determine divergence dates and phylogenetic relationships revealed unique FVs with a divergence date of approximately 60 million years. This study helps us understand the FVs evolution and provides a scientific basis for future investigations into animal-borne infectious diseases.

Complete genome sequence of Valeriana jatamansi cryptic virus 1: a novel member of the genus Alphapartitivirus infecting Valeriana jatamansi Jones.

A new double-stranded RNA (dsRNA) virus, tentatively named "Valeriana jatamansi cryptic virus 1" (VJCV1, GenBank accession nos. PP482519 and PP482520), was isolated from diseased Valeriana jatamansi Jones plants exhibiting vein-banding in Yunnan. Its complete genome sequence was determined using metatranscriptomic and Sanger sequencing. The genome of VJCV1 consists of two dsRNA of different size, namely dsRNA1 (2,026 bp) and dsRNA2 (1,754 bp), which are predicted to encode an RNA-dependent RNA polymerase (RdRp, 616 aa) with molecular weight of 72.6 kDa and coat protein (CP, 491 aa) with molecular weight of 55.8 kDa, respectively. The non-coding region of dsRNA in VJCV1 is predicted to have a stem-loop structure and a poly(A) tail that are unique to the members of the genus Alphapartitivirus. Multiple sequence alignments showed that the RdRp and CP of VJCV1 shared the highest amino acid sequence identity (86.2% and 56.1%, respectively) with red clover cryptic virus 1 (RCCV1). These values are below the threshold for creating new species within the genus Alphapartitivirus. Phylogenetic analysis based on RdRp and CP sequences showed that VJCV1 clustered independently from members of the genus Alphapartitivirus, with RCCV1 being the closest relative. It is therefore suggested that VJCV1 should be considered a member of a new species of the genus Alphapartitivirus in the family Partitiviridae. This is the first report of a member of the genus Alphapartitivirus infecting a plant of the genus Valeriana.

The complete chloroplast genome of a centennial olive tree (Olea europaea, Oleaceae) from the southern Peruvian coast

Olive (Olea europaea Linaeus 1753) is one of the valuable fruit trees and very important edible oil plant in the world. The chloroplast (cp) genome of an olive tree (Olea europaea) from the southern Peruvian arid coast was obtained for the first time. Genomic DNA of high quality was used to generate librarieswith Illumina Hiseq paired-end methods. The cp genome is 155,886 pb in length and contains a large single-copy region (LSC) of 86,610 pb and a small single copy region (SSC) of 17,790 pb separated by two inverted repeat (IR) regions (25,741bp). The cp genome of olive contains 124 genes that consists of 80 protein-coding genes, 36 tRNA, eight rRNA. Phylogenetic analysis showed this olive tree is sister to O. europaea subsp. maroccana (Oleeae tribe). This study presents the first overview of the chloroplast genome organization and phylogenetics of O. europaea, offering valuable insights for genetic and evolutionary research in the genus Olea.

Locust Pathogen Aspergillus oryzae XJ1 Is Different from Aspergillus oryzae and Aspergillus flavus Based on Genomics Comparisons

Fungi play an increasingly important role in the biological control of insect pests. Aspergillus oryzae XJ1 is highly virulent to locust adults and nymphs, which are a destructive economic pest worldwide. Because of its host association with locusts, which is unique in Aspergillus, in this study, we examined the genetic relationships of A. oryzae XJ1 within Aspergillus. We sequenced the genome of A. oryzae XJ1 and compared it with the genomes of other Aspergillus species. The complete genome of A. oryzae XJ1 is 37.9 Mb, comprising 11,720 putative genes, assembled into eight chromosomes. The genome size is similar to that of other A. oryzae strains. Phylogenomic analysis indicated that A. oryzae XJ1 was most closely related to A. flavus NRRL3357, not A. oryzae RIB40. Core/pan-genome analysis of A. oryzae XJ1 and other Aspergillus species revealed that A. oryzae XJ1 had 704 strain-specific genes, whereas A. flavus NRRL3357, A. oryzae KDG 21, and A. parasiticus NRRL 2999 had 646, 955, and 779 unique genes, respectively. The A. oryzae XJ1 genome showed structural differences compared with the genomes of A. oryzae RIB40 and A. flavus NRRL3357 in genomic synteny analysis. These results indicate that A. oryzae XJ1 is genetically distinct at the genome level from other Aspergillus species, including A. oryzae and A. flavus, and may be as a distinct species. This will provide new insight into the classification of Aspergillus based on genomics.

Severe fever with thrombocytopenia syndrome virus was found in Northern Jiangxi Province, China

IntroductionSevere fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease discovered in China in 2009. SFTS monitoring has been carried out since 2010 in mainland China. In recent years, human infection with SFTS virus (SFTSV) has frequently been detected in Jiujiang of Jiangxi Province, Central China.MethodsSera of SFTS surveillance cases and samples collected from humans, animals and ticks surrounding the cases were used to detect SFTSV RNA by real-time RT-PCR. SFTSV-positive samples were further subjected to sequencing and analysis of the S, M, and L segments of SFTSV.ResultsFour patients were positive for SFTSV infection. However, the subjects like humans, animals and ticks around the cases were all detected as negative for the virus infection. Phylogenetic analysis of the partial segments revealed that SFTSVs in three patients from Jiujiang were clustered with genotype C4 and C5 on the L and M segment-established phylogenetic tree; however, phylogenetic analysis of the S segment showed that the strains were grouped into genotype J1. These results suggested that the S, M, and L segments of these strains underwent segmental reassortment, which was later supported by recombination signal detection. The reassortment event was detected by at least four methods with a significance level of p value <0.05. In addition, the RDP recombination consensus score (RDPRCS) was greater than 0.40. To avoid poor tree topology support with the partial S, M, and L segments, we further performed analysis of the complete genome of SFTSV. The full-length L, M, and S segment sequences of the strains were consistently clustered into two genotypes, namely the genotype C5 and the genotype C4. The strains belonging to genotype C5 were detected for recombination signal with moderate confidence by all the six methods, with a significance level of p value <0.05 and an RDPRCS of 0.41. The recombination event might have occurred between the minor parent (2011YPQ11, C3/C3/C3) and major parent (SPL053A, J1/ J1/ J1). However, there was no genetic recombination detected in the strains belonging to genotype C4. The event was detected by only two methods with a significance level of p value <0.05.DiscussionTwo genotypes of SFTSVs were identified in Jiujiang City, Jiangxi Province of Central China, and they were genotype C5 and genotype C4. The genotype C5 underwent genetic recombination in this region, with the minor parent of the strain 2011YPQ11 and the major parent of the strain SPL053A.

Comparative and Adaptive Analyses of the Complete Chloroplast Genome Diversity in Sium serra

Background/Objectives: Sium serra is distributed in Korea, China, and Japan. It was first identified as the genus Pimpinella and then reclassified as Sium by Kitagawa. Some Sium species are used as herbal medicine and are often confused with the similar form Ligusticum sinense. In this study, we analyzed the cp genome of S. serra and conducted comparative analyses with the cp genomes of related taxa. Methods: We extracted gDNA from fresh leaves and sequenced it using Illumina HiSeq2500. For the chloroplast genome assembly, de novo assembly was performed using Velvet v1.2.07. For the annotation, GeSeq and NCBI BLASTN were used. Afterwards, related taxa were analyzed using programs such as DnaSP and MISA. Results: S. serra was excluded from the study on the chloroplast (cp) genome in Sium because it was classified as Pimpinella in China. Therefore, this study aimed to analyze the cp genome of S. serra for the first time and its location within the genus Sium. The complete cp genome of S. serra was 154,755 bp in length, including a pair of inverted repeats, each 26,255 bp, a large single-copy region of 84,581 bp, and a small single-copy region of 17,664 bp. The cp genome comprised 79 protein-coding, 30 tRNA, and 4 rRNA genes. Furthermore, six regions of high nucleotide diversity were identified in the genus Sium. In the genus Sium, 1630 repeats that can serve as markers were also identified. Eight protein-coding genes with high KA/KS values were under positive selection in the Sium. Our phylogenetic analyses suggest that S. serra was positioned with high bootstrap support within the Sium of the tribe Oenantheae, specifically in the southern Palearctic subclade. Conclusions: In this study, the S. serra chloroplast genome was sequenced and assembled. The genus Sium formed a monophyletic group; however, as not all the Sium species were included in this study, further research is necessary. This study can serve as foundational data not only for Sium but also for the tribe Oenantheae.

Mitochondrial Genomes of Mammals from the Brazilian Cerrado and Phylogenetic Considerations for the Orders Artiodactyla, Carnivora, and Chiroptera (Chordata: Mammalia)

We assembled and annotated the complete mitochondrial genomes of Lycalopex vetulus (hoary fox), Cerdocyon thous (bush dog), Tayassu pecari (white-lipped peccary), and Tadarida brasiliensis (Brazilian free-tailed bat). The mitogenomes exhibited typical vertebrate structures, containing 13 protein-coding genes, 22 tRNA genes, 2 ribosomal RNA genes, and a D-loop region. Phylogenetic reconstruction using the 13 protein-coding genes revealed robust relationships among species within Carnivora, Chiroptera, and Artiodactyla, corroborating previous studies. Secondary structure analysis of tRNAs and ribosomal genes showed slight variations among species of the same order. This research highlights the importance of mitochondrial genomics in understanding the evolutionary relationships and genetic diversity of Cerrado mammals, contributing to conservation efforts for this unique ecosystem.

Analysis of the mitochondrial genome to determine the origins and pathways of entry of Angiostrongylus cantonensis in continental Europe (Valencia, Spain)

Abstract Angiostrongylus cantonensis, the rat lungworm, is a zoonotic parasite mainly of rats which act as definitive hosts. If humans become accidentally infected, the nematode is capable of migrating to the brain causing meningoencephalitis. Intermediate hosts are snails and slugs. Although originating from mainland China, A. cantonensis has now spread to various countries and continents. The precise timing of its departure from mainland China remains uncertain although it is often associated with significant historical events or migratory movements. The exit of A. cantonensis from mainland China is believed to have occurred in a singular event, followed by its divergence into 2 distinct clades: clade I, originating from mainland China, and clade II, representing global spread. Angiostrongylus cantonensis was first identified in continental Europe in 2021, specifically in Valencia, Spain. Illumina genome sequencing of 7 individuals isolated from rats captured in 2 different districts in the city of Valencia was carried out. The complete mitochondrial genome was assembled and compared with published A. cantonensis mitochondrial genomes through Bayesian phylogenetic analysis, both for complete mitochondrial genomes and for the cytochrome c oxidase I gene, given its widespread use for identification of the species. The findings revealed the presence of 2 different A. cantonensis haplotypes in the rats studied in Valencia, both belonging to clade II. In 2 rats both clades were present.

A retrospective genomic characterisation of the 2022 mpox outbreak in Belgium, and in vitro assessment of three antiviral compounds

Since the beginning of May 2022, cases of mpox have been reported in several European and American countries where the disease is nonendemic. We performed a retrospective genomic characterisation of the 2022 mpox outbreak in Belgium, and assessed the invitro sensitivity of three antiviral compounds to a monkeypox virus (MPXV) strain from the 2022 outbreak. We sequenced the complete genomes of MPXV isolated from skin-, throat-, anorectal- and genital swab samples using the Oxford Nanopore Technologies (ONT) GridION. We subsequently analysed high-quality complete MPXV genomes and conducted a genomic analysis of MPXV complete genomes from this study with all other complete MPXV genomes available on GISAID up to October 28th, 2022. The invitro activity of tecovirimat, brincidofovir, and cidofovir was also tested in human and monkey cell lines. We produced 248 complete MPXV genomes. Phylogenetic analysis of the complete MPXV genomes revealed that they all belong to MPXV Clade IIb B.1. Surprisingly, through phylogeographic analysis we identified a minimum number of 79 introduction events into Belgium, along with sustained local transmission. We also demonstrated the superior invitro efficacy and selectivity of tecovirimat to the 2022 MPXV clinical strain. The number of sequences provides sufficient information about the MPXV lineages that were circulating in Belgium. The 2022 mpox outbreak, in Belgium, was mainly characterised by many introduction events that were promptly contained and resulted in limited human-to-human transmission of MPXV. The invitro efficacy of antivirals against a 2022 MPXV Belgian strain highlights the potent activity and specificity of tecovirimat and its ability to prevent the formation of the extracellular enveloped viruses. None.

The complete mitochondrial genome of Dendrodoris krusensternii (Gastropoda, Nudibranchia, Dendrodorididae) from South Korea

The complete mitochondrial genome sequence of Dendrodoris krusensternii (J. E. Gray, 1850) was determined using next-generation sequencing. The complete mitochondrial genome of D. krusensternii is 14,361 bp long, comprising 13 genes, including 13 protein-coding genes (PCGs), 22 transfer RNAs, and two ribosomal RNAs. The nucleotide composition was estimated: 28.7% A, 14.8% C, 19.6% G, and 36.9% T. Phylogenetic analysis was performed using the maximum likelihood method (including 13 PCGs). D. krusensternii is related to the Phyllidiidae (superfamily Phyllidioidea), suggesting a distinct phylogenetic placement within Nudibranchia. This study represents a genomic resource, contributing to molecular studies on the evolution of the Dendrodorididae.

The complete chloroplast genome sequence of Melothria scabra (Cucurbitaceae)

Melothria scabra has gradually become an economically important plant worldwide. The complete chloroplast genome of M. scabra has a length of 156,744 bp, contains a large single-copy (LSC) region (86,387 bp), a small single-copy (SSC) region (18,055 bp), and two inverted repeats (IRs) with the same length of 26,151 bp. In total, 126 genes were detected, including 83 protein-encoding genes, 35 transfer RNA (tRNA) genes, and eight ribosomal RNA (rRNA) genes. For phylogenetic analysis, M. scabra has a closer genetic relationship with Cucumis sativus and Citrullus lanatus. The complete chloroplast genome sequence of M. scabra would promote the germplasm exploration, phylogenetic relationships, and molecular biology researches in Melothria.

Diverse genomic and epidemiological landscapes of redundant pbp5 genes in Enterococcus spp.: Insights into plasmid mobilization, ampicillin susceptibility, and environmental interactions.

Genetic redundancy in bacteria plays a crucial role in enhancing adaptability and accelerating evolution in response to selective pressures, particularly those associated with rapid environmental changes. Aminopenicillins like ampicillin are important therapeutic options for Enterococcus infections in both humans and animals, with resistance mostly associated with pbp5 gene mutations or overexpression. While the occurrence of redundant pbp5 genes has been occasionally reported, the advantages for the host bacteria have not been explored in detail. During a whole-genome sequencing project of Enterococcus faecium from bacteremic patients, we identified an ST592 strain (Efm57) with redundant pbp5 genes. This presented an opportunity to investigate the prevalence and implications of multiple pbp5 acquisitions in diverse Enterococcus species across various sources, geographical regions, and timeframes. The analysis of 618 complete Enterococcus genomes from public databases revealed that 3.2% harbored redundant pbp5 genes, located on chromosomes or plasmids across different species from diverse epidemiological backgrounds. The proteins encoded by these genes showed homologies ranging from 51.1% to 97.5% compared to native copies. Phylogenetic analysis grouped redundant PBP5 amino acid sequences into three distinct clades, with insertion sequences (mostly IS6-like) facilitating their recent spread to diverse plasmids with varying genetic backbones. The presence of multiple antibiotic resistance genes on pbp5-plasmids, including those conferring resistance to linezolid, underscores their involvement in co-selection and recombination events with other clinically-relevant antibiotics. Conjugation experiments confirmed the transferability of a specific 24kb pbp5-plasmid from the Efm57 strain. This plasmid was associated with higher minimum inhibitory concentrations of ampicillin and conferred bacteria growth advantages at 22°C. In conclusion, the widespread distribution of redundant pbp5 genes among Enterococcus spp. highlights the complex interplay between genetic mobility, environmental factors, and multidrug resistance in overlapping ecosystems emphasizing the importance of understanding these dynamics to mitigate antibiotic resistance spread within the One Health framework.