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. 2026 Feb 24;12:101362. doi: 10.1016/j.crfs.2026.101362

First co-emergence of plasmid-borne tigecycline resistance genes tet(X4) and tmexCD1-toprJ1 in Raoultella ornithinolytica isolated from retail pork: insights from global genomic analysis

Muhammad Shoaib a,1, Asim Munir a,1, Muhammad Fazal Hameed a, Shah Zeb a, Zhichao Li a, Kai Peng a, Nishant Shah a,b, Patrick Butaye c,d, Zhiqiang Wang a,⁎, Ruichao Li a,⁎⁎
PMCID: PMC12969439  PMID: 41810423

Abstract

The emergence and spread of resistance to last-resort antibiotics such as tigecycline is of great concern globally which can pose a potential threat to public health due to limited treatment options. In this study, we first identified and characterized the genomic features of a food-acquired Raoultella ornithinolytica strain co-harboring tet(X4) and tmexCD1-toprJ1 genes through antibiotic susceptibility testing and whole genome sequencing, followed by global comparative analysis of Raoultella ornithinolytica genomes submitted in the National Center for Biotechnology Information (NCBI) database. Raoultella ornithinolytica LYG_6_1B exhibited high-level tigecycline resistance and multi-drug resistance. The genomic analysis revealed that LYG_6_1B harbored 32 antibiotic resistance genes (e.g., tetracycline, sulfonamides, trimethoprim, aminoglycosides, fluoroquinolone, phenicol, β-lactam, macrolides, and rifampicin), 14 virulence genes (e.g., ecpA, ompA, entB, irp1, irp2, ybtAEPQSTUX, fyuA, entB), and three plasmids. The tet(X4) was found to be located on 79 Kb IncFII(pCRY) plasmid and tmexCD1-toprJ1 on 290 Kb hybrid plasmid [IncFIB(pNDM-Mar) + IncHI1B(pNDM-Mar)]. Both of the genes, tet(X4) and tmexCD1-toprJ1 were found to be located on separate conjugative plasmids via the conjugation assay. This indicates the expanding host range and independent horizontal transfer potential of these critical resistance genes across bacterial species or genera in One Health settings. Therefore, the current study recommends the continuous genomic surveillance of bacterial strains in food supply chain and other One Health settings to monitor and prevent their further dissemination. Moreover, the emergence of critical resistance genes in food-origin bacteria can pose a direct threat to human health.

Keywords: Raoultella ornithinolytica, Retail pork, tet(X4), tmexCD1-toprJ1, Genomic analysis

Graphical abstract

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Highlights

  • •

    This study first identified and provides the genomic characteristics of tet(X4) and tmexCD1-toprJ1 in R. ornithinolytica strain from a retail pork.

  • •

    The tet(X4) and tmexCD1-toprJ1 harboring plasmids were transferable and may have the potential to spread globally among One Health components.

  • •

    Global genomic analysis revealed the potential risks to animal, human, and environmental health.

1. Introduction

Raoultella (R.) ornithinolytica is a reclassified (formerly known as Klebsiella ornithinolytica) encapsulated Gram-negative bacterium belonging to Enterobacteriaceae family (Drancourt et al., 2001). Nowadays, R. ornithinolytica has been recognized as an emerging opportunistic pathogen in humans. Its clinical significance has grown with increased reporting of infections, particularly in the healthcare settings and among immunocompromised patients (Büyükcam et al., 2019; Seng et al., 2016). Previously, multiple cases of urinary tract infections, bacteremia, and biliary tract infections caused by R. ornithinolytica have been reported (Seng et al., 2016). R. ornithinolytica infections can be under looked because of difficult identification using phenotypic methods, however, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technology has overcome this issue (de Jong et al., 2013).

Moreover, the growing antimicrobial resistance (AMR) in One Health sectors poses a significant threat to the public health. The interconnection between different reservoirs such as food, animal, human, and environment can play a significant role in the emergence and spread of AMR through diverse bacterial species (Martak et al., 2024; Wang et al., 2026). The recent emergence of multi-drug resistant (MDR) Enterobacterales such as carbapenem-resistant, colistin resistant, and extended-spectrum beta-lactamase producing bacteria has significantly compromised the efficacy of carbapenems, colistin, and cephalosporins, respectively (Cai et al., 2026; Shoaib et al., 2024; Wu et al., 2024). Moreover, the prevalence of carbapenem-resistance in R. ornithinolytica has also been reported increasingly with fewer reports of colistin resistance (Li et al., 2024; Wang et al., 2019).

Tigecycline is a last-resort third-generation antibiotic belong to tetracycline class exhibits an extended-spectrum activity to treat MDR bacterial infections including caused by carbapenem- and colistin-resistant bacteria (Ortega-Balleza et al., 2026). Worryingly, the recent emergence of plasmid-mediated tigecycline resistance tmexCD-toprJ cluster in R. ornithinolytica from animal, environment, and clinical sources has raised the potential concerns (Li et al., 2024; Wang et al., 2021). Moreover, the plasmid-mediated tet(X) variants have also been circulating in diverse bacteria species (Rueda Furlan et al., 2023). The plasmid-mediated tigecycline resistance genes can pose a potential threat and critical challenge to public health by inactivating the last-resort antibiotic and dissemination globally across bacterial species or genera through horizontal gene transfer (Yao et al., 2025). The major threat is co-existence with carbapenemase and colistin resistance genes within same bacterium leading to the emergence of pan-resistant “Superbugs” (Mataseje Laura et al., 2025; Wang et al., 2025).

Such genes have been reported in diverse bacteria but there are fewer or no reports in R. ornithinolytica. In this study, we have identified and characterized a R. ornithinolytica strain LYG_6_1B co-carrying tet(X4) and tmexCD1-toprJ1 genes from retail pork in Jiangsu, China. We have also performed the comparative analysis with global R. ornithinolytica strains submitted in the National Center for Biotechnology Information (NCBI) (accessed on April 1, 2025) database. Based on the analysis, we have first identified tet(X4) in R. ornithinolytica of food-origin. However, the tmexCD1-toprJ1 has been reported in R. ornithinolytica from other sources but we are reporting it for the first time from food-source.

2. Materials and methods

2.1. Bacterial isolation and identification

Between September 2024 to February 2025, seventy-two pork samples were collected from retail markets from different cities in Jiangsu province, China. The samples were processed following the earlier study protocol (Li et al., 2023). Briefly, the 5g of pork sample was enriched in 100 mL Luria-Bertani (LB) broth containing 4 μg/mL tigecycline for 6 h at 37°C. Then, the enriched broth was inoculated on LB agar supplemented with tigecycline (4 μg/mL) to only select tigecycline-resistant isolates. The 2-3 representative isolated colonies were picked and further inoculated until purified. The purified colony was used for species identification by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (Bruker, Massachusetts, USA).

2.2. Antimicrobial susceptibility testing

The antimicrobial susceptibility was done by the broth microdilution assay to determine the minimum inhibitory concentration (MIC) of eight antimicrobial agents including tigecycline (TGC), meropenem (MEM), colistin sulfate (CS), ciprofloxacin (CIP), florfenicol (FFC), fosfomycin (FOS), gentamicin (GEN), and ceftriaxone (CFX) following the Clinical and Laboratory Standards Institute (CLSI) guidelines (CLSI, 2024). Briefly, the bacterial culture was grown overnight. The culture was then adjusted to meet the 0.5 McFarland standard and then diluted to achieve the final concentration of approximately 5 × 105 CFU/mL in 96-well plates containing fresh cation-adjusted Mueller-Hinton (MH) broth and serially diluted antibiotics. The plates were incubated at 37°C for 16-20 h and the MIC results were noted visually. The results were interpreted according to the breakpoints established by CLSI except for tigecycline, which was interpreted according to the European Committee on Antimicrobial Susceptibility Testing (v.14.0) guidelines (EUCAST, 2024).

2.3. Whole genome sequencing, data extraction, and bioinformatics analysis

The genomic DNA of R. ornithinolytica strain LYG_6_1B was extracted using the Vazyme Bacterial DNA Kit (Vazyme, Beijing, China) and quantified using the Nandrop technology (Thermofisher Scientific, USA). The whole genome sequencing (WGS) was performed on the NovaSeq 6000 (Illumina, CA, USA) for short-reads and MinION (Nanopore, Oxford, UK) for long-reads. The short-read raw data obtained were trimmed for low quality-reads and adaptor removal using the Trimmomatic v.0.39 (Bolger et al., 2014) with parameters: ILLUMINACLIP:TruSeq3-PE.fa:2:30:10 LEADING:3 TRAILING:3 SLIDINGWINDOW:4:15 MINLEN:36 and further quality check by FastQC v.0.11.8 (Wingett and Andrews, 2018) with default parameters. The de novo hybrid assembly of short-reads and long-reads were generated using the Unicycler v0.5.1 with default parameters. Moreover, all R. ornithinolytica genome sequences and their metadata were downloaded from NCBI using the datasets command-line tool (accessed on April 1, 2025). The genome data were processed for contamination quality check using the CheckM2 v1.0.2 (https://github.com/chklovski/CheckM2) with default parameters. The assembled genome of this study strain and other from public database were processed to identify resistome, plasmids, and virulome using the NCBI AMRFinderPlus, PlasmidFinder, and Virulence Factor Database (VFDB) in the ABRicate package v.1.0.1 (https://github.com/tseemann/abricate) using the default parameters (minimum identity 80%, minimum coverage 80%). The genome annotation was done by Prokka v.1.14.6 (Seemann, 2014) using the default parameters (e-value 1e-06, minimum coverage 80%, kingdom Bacteria) and the alignment was constructed using the Roary v.3.13.0 (Page et al., 2015) with default parameters (BLASTp identity 98%, core definition 99%) based on core-genome single nucelotide polymorphisms (cgSNPs). The final phylogenetic tree was constructed using the FastTree v.2.1.11 (Price et al., 2010) with default parameters (Jukes-Cantor nucleotide evolution model, CAT approximation for rate heterogeneity).

2.4. Plasmid mobility testing

The mobility of plasmids was first predicted in-silico by Mob-Suite online server (https://www.solugenomics.com/tools/mob-suite) by simply uploading the genome fasta file. Further, in-vitro conjugation assay was performed to assess the transferability of the plasmids harbouring tet(X4) and tmexCD1-toprJ1 in LYG_6_1B donor to the recipient E. coli J53 (sodium azide resistance) following the earlier protocol (Zhang et al., 2025). Briefly, the donor and recipient bacteria were grown up-to mid-log phase (an optical density of 0.8 at 600 nm) and then mixed at a 1:1 ratio. The mixed cultures were incubated overnight at 37°C and then 10-fold serially diluted followed by plating on selective LB agar plates (Oxoid, UK) supplemented with tigecycline (4 μg/mL) and sodium azide (200 μg/mL) for transconjugants and plates containing only sodium azide (200 μg/mL) for recipient cells. The number of transconjugants and recipient cells were determined by counting the colonies grown on selective plates. The conjugation frequency was determined by the ratio of transconjugants to recipient cells. Transconjugants were confirmed by conventional PCR and Sanger sequencing for the presence of tet(X4) and tmexCD1-toprJ1 genes.

2.5. Data visualization

The phylogenetic tree, metadata, resistome, virulome, plasmid types, and bar graphs visualization were done on the Interactive Tree of Life (iTOL) web-based server (https://itol.embl.de/, accessed on August 20, 2025). The circular comparison of plasmids from this study and other plasmids from public database were done using the BLAST Ring Image Generator (BRIG) version 0.95 (Alikhan et al., 2011). The genome mapping was done using the gbdraw v0.8.0 (https://github.com/satoshikawato/gbdraw, accessed on October 15, 2025) online.

3. Results and discussion

3.1. Global phylogeny and epidemiological analysis of R. ornithinolytica

In this study, we isolated one tigecycline resistant R. ornithinolytica strain LYG_6_1B (MIC = 128 μg/mL) from retail pork. The observed MIC of tigecycline found to be noted much higher than the levels reported individually in previous studies (Cui et al., 2022; He et al., 2019). The higher MIC exhibited by current study strain might be due to synergistic interaction between two co-localized determinants. The enzymatic inhibition of tigecycline by tet(X4) in combination with active extrusion of antibiotic by TmexCD1-ToprJ1 efflux pump likely resulted in the reduced intracellular drug accumulation, therefore exceeding the additive impact of individual genes (Su et al., 2024). However, further studies are required to confirm this phenomenon. The MIC level can also far exceed due to the effect of sub-inhibitory concentrations of tigecycline and can lead to persistence of tigecycline resistance after selection (Liu et al., 2023). Further, global genomic analysis of 568 R. ornithinolytica strains from NCBI database revealed only 12/568 (2.1%) R. ornithinolytica strains carrying tmexCD-toprJ gene cluster while none of them carried the tet(X) gene. Global phylogeny of 569 R. ornithinolytica strains revealed that our study strain showed close similarity with two environmental strains, 2SHCen5mer (GCF_046602315.1) and R. ornithinolytica strain 18 (GCA_001723565.1) from Czech Republic and China in 2025 and 2016, respectively. Surprisingly, the other similar strains in this clade were all recovered from Homo sapiens in Germany, Russia, and Ecuador and none of the strain carried tigecycline resistance gene (Fig. 1).

Fig. 1.

Fig. 1

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Global phylogeny of R. ornithinolytica strains submitted in the NCBI database (accessed on April 1, 2025) and from this study (LYG_6_1B). Phylogenetic tree was constructed based on core-genome single nucleotide polymorphisms (cgSNPs) alignment using the Roary v.3.13.0. Different branch colors indicate different major clades while the year and isolation source are presented as text labels. Country and host information is presented as color strips while the tet(X) and tmexCD-toprJ absence and presence is presented as filled and hollow circles.

3.2. Genomic analysis of tet(X)- or tmexCD-toprJ-carrying R. ornithinolytica strains

The hybrid assembly of LYG_6_1B strain showed that the complete genome consisted of a chromosome of 5,643,340 bp and three plasmids designated as pLYG_6_1B_111 Kb, pLYG_6_1B_290 Kb, and pLYG_6_1B_79 Kb (Fig. 2a-d). Further, we performed the phylogeny, epidemiological, and resistome analysis of 12 public strains and LYG_6_1B identified in this study (Fig. 3A). The comprehensive resistome analysis showed that LYG_6_1B strain harbored 32 antibiotic resistance genes (ARGs) including tigecycline [tet(X4), tmexCD1-toprJ1], tetracycline [tet(A)], sulfonamides (sul1, sul2, sul3), trimethoprim (dfrA12, dfrA27), aminoglycosides [aac(3)-IVa, aph(3′)-Ia, aph(3″)-Ib, aph(4)-Ia, aph(6)-Id, aadA1, aadA2, aadA16], fluoroquinolone (qnrS1, qnrB4, oqxA10, oqxB9), phenicol (cmlA1, floR), β-lactam (blaORN-3, blaDHA-1), fosfomycin (fosA), macrolides [estT, mph(E), msr(E), armA], and rifampicin (arr-3). Similarly, other strains were also carrying higher number of ARGs (≥21) except NC189 (GCA_016618175.1) isolated from clinical settings in China (Fig. 3A). LYG_6_1B was also found to possess multiple plasimds and virulence genes (VGs) similar to other tmexCD-toprJ-positive strains (Fig. 4). The identification of 32 ARGs confer resistance to ≥3 antimicrobial classes in a single strain highlight its potential impact for multi-drug-resistance. Moreover, the identification of 14 VGs are of high clinical significance due to the bacterial adhesion/colonization (e.g., ecpA, ompA, entB) with host cells and iron acquisition (e.g., irp1, irp2, ybt cluster, fyuA, entB) through them. The ecpA gene is a component of ecpRABCDE operon which encode for the thin-structures called as pilli or fimbriae that allow the bacterium to attach to host epithelial cells (Rendón et al., 2007). ompA is an outer membrane protein A that performs multiple functions including the integrity of bacterial cell membrane, adhesion and invading the host cells, and evading host immune defenses (Confer and Ayalew, 2013). entB gene is primarily a part of enterobactin system, a major siderophore for iron acquisition but can also acts as signaling molecule for biofilm formation (Amiri et al., 2025). Moreover, in the iron acquisition system, irp1 and irp2 act as iron regulatory proteins throughsynthesis of iron-scavenging molecule called as yersiniabactin. The ybtAEPQSTUX cluster facilitates the yersiniabactin synthesis, regulation, and transport. The fyuA gene encodes for yersiniabactin receptor that recognizes and imports the iron-acquired yersiniabactin molecule back into the bacterial cell (Gravey et al., 2024). The co-existence of potent virulence arsenal with multi-drug resistance in a food-origin bacterial strain is a serious clinical concern.

Fig. 2.

Fig. 2

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Complete genome map of LYG_6_1B strain. (a) Chromosome, the scale indicates the ribosomal RNA (rRNA), transfer RNA (tRNA), coding sequences (CDS), non-coding (ncRNA), transfer-messenger RNA (tmRNA), repeat regions, GC content, and GC Skew. (b) Plasmid pLYG_6_1B_79 Kb, the scale indicates the coding sequences (CDS), GC content, and GC Skew. (c) Plasmid pLYG_6_1B_290 Kb, the scale indicates the coding sequences (CDS), GC content, and GC Skew. (d) Plasmid pLYG_6_1B_111 Kb, the scale indicates the coding sequences (CDS), GC content, and GC Skew.

Fig. 3.

Fig. 3

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(A) Phylogeny, epidemiological, and antibiotic resistance genes (ARGs) distribution among the tet(X)- or tmexCD-toprJ cluster-carrying R. ornithinolytica strains. The blue scale bar graph represents the No. of ARGs acquired by each strain. (B) Circular comparison between tmexCD1-toprJ1-bearing plasmid (pLYG_6_1B_290 Kb) found in this study and all other tmexCD-toprJ gene cluster carrying R. ornithinolytica plasmids. The outermost red circle represents the annotation features and reference plasmid (pLYG_6_1B_290 Kb). The plasmid pM27-NDM-363K (CP130154.1, 363,911 bp) carrying tmexCD2-toprJ2 cluster from R. ornithinolytica strain RoM27LC23 showed highest identity and coverage. (C) Circular comparison between tet(X4)-bearing plasmid (pLYG_6_1B_79 Kb) found in this study and other closely-related plasmids in the NCBI database. The outermost green circle represents the annotation features and reference plasmid pLYG_6_1B_79 Kb. The two plasmids pL3995-3 (CP135167.1, 78,154 bp) and pNTT31XS-tetX4 (CP077430.1, 78,159 bp) identified in K. pneumoniae and K. aerogenes from human stool and pig intestinal content in China showed 100% nucleotide identity and 99% coverage with pLYG_6_1B_79 Kb plasmid from this study, respectively.

Fig. 4.

Fig. 4

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Phylogeny, distribution of plasmid replicon types (PRTs), and virulence genes (VGs) across the tet(X)- or tmexCD-toprJ-harboring R. ornithinolytica strains. Red label strain in the phylogenetic tree identified in this study. The color strip indicates the year of reporting. Purple and dark yellow binary map represents the presence and absence of plasmid types and VGs, respectively. The purple and green bar graphs represent the No. of plasmids and No. of VGs carried by each strain, respectively.

3.3. Plasmid analysis of tet(X)- or tmexCD-toprJ-bearing plasmids

The plasmid analysis revealed that tmexCD1-toprJ1 resistance cluster along with other MDR genes were found to be located on pLYG_6_1B_290 Kb hybrid plasmid [IncFIB(pNDM-Mar) + IncHI1B(pNDM-Mar)] while tet(X4) with estT, catB, and marR were carried by pLYG_6_1B_79 Kb belonging to IncFII(pCRY) type (Fig. 3A & B). Third plasmid, pLYG_6_1B_111 Kb was found to carry terB and dfr genes which confer resistance to tellurite and trimethoprim, respectively (Fig. S1).

The complete plasmids carrying tmexCD-toprJ gene cluster in R. ornithinolytica were downloaded from the NCBI and compared in this study. The plasmid analysis revealed that pLYG_6_1B_290 Kb showed highest identity and coverage with pM27-NDM-363K (CP130154.1, 363,911 bp) mega-plasmid carrying tmexCD2-toprJ2 cluster which was identified in R. ornithinolytica strain RoM27LC23 from garbage bins in Shandong, China (Li et al., 2024). The genetic context of tmexCD1-toprJ1 cluster in the pLYG_6_1B_290 Kb (IS26-aph(3′)-Ia-IS26-tmexC1-tmexD1-toprJ1-aph(3″)-Ib-△IS903-IS26) was also found different than pM27-NDM-363K and other plasmids. All other plasmids lack the insertion of aph(3′)-Ia while pM27-NDM-363K lacks the insertion of aph(3″)-Ib-△IS903 in their backbone (Fig. 3B). The insertion of mobile element (e.g., IS903) and ARG (e.g., aph(3′)-Ia) into the plasmid backbones near the key resistance genes is a classic sign of ongoing modular evolution and adaptation, increasing their stability and co-selection potential (Khedkar et al., 2022; Partridge et al., 2021).

BLASTn search of tet(X4)-carrying plasmid revealed only four plasmids with ≥99% identity and ≥91% query coverage. All other similar plasmids in NCBI exhibited ≤87% coverage and 99.99% identity predominately identified in Klebsiella spp (Table S1). The two plasmids pL3995-3 (CP135167.1, 78,154 bp) and pNTT31XS-tetX4 (CP077430.1, 78,159 bp) identified in K. pneumoniae and K. aerogenes from human stool and pig intestinal content in China showed 100% nucleotide identity and 99% coverage with pLYG_6_1B_79 Kb plasmid from this study, respectively. The genetic context of the tet(X4) was found similar in both plasmids but the insertion of IS903 was lacking in all other similar plasmids. The pLYG_6_1B_79 Kb and pYZ22PK101_3 (CP114027.1, 78,159 bp) showed 99.99% identity and 99% coverage with similar genetic environment around tet(X4) gene. The pYZ22PK101_3 plasmid was identified in K. quasipneumoniae strain YZ22PK101 from pork in Yangzhou, China. Another plasmid, pYZ22PK094_1 (CP114017.1, 110,961 bp) also showed 100% identity and 91% coverage but surprisingly lacking the tet(X4) and other (estT, catB, marR) genes while the ISCR2 sequences lies in its backbone (Fig. 3C).

3.4. Plasmid mobility analysis

The mobility of plasmids was first predicted by Mob-Suite (https://www.solugenomics.com/tools/mob-suite) and final confirmation by conjugation assay. The Mob-Suite results showed that pLYG_6_1B_290 Kb and pLYG_6_1B_79 Kb are both conjugative carrying mobH and mobP relaxase genes, respectively. The mobilization (MOB) genes play an important role in the plasmid typing and elucidate their transmission mechanism. The identification of different MOB types indicates their distinct transmission mechanism, emergence of resistance traits, adaptation and evolution of bacteria (Garcillán-Barcia et al., 2009, 2011). Further, mate-pair forming (MPF) typing revealed that pLYG_6_1B_290 Kb plasmid carried the MPF_F protein while pLYG_6_1B_79 Kb plasmid carried the MPF_I protein. These proteins mediates the contact and DNA exchange between donor and recipient strains during conjugation (Shintani et al., 2015). The final confirmation by conjugation assay revealed the transfer of tet(X4) and tmexCD1-toprJ1 into the recipient E. coli J53 with a conjugation efficiency of 5.42 ± 1.56 × 10−4 and 2.79 ± 2.14 × 10−6, respectively. The higher efficiency of tet(X4)-bearing plasmid might be due to small size and single plasmid type while the presence of tmexCD1-toprJ1 on hybrid-type and large plasmid can lead to lower conjugation efficiency. Moreover, the identification of tmexCD1-toprJ1 and tet(X4) genes on separate conjugative plasmids allows independent dissemination and potential re-assortment in bacterial populations, creating “super-resistant” strains (Li et al., 2024; Yao et al., 2024).

4. Conclusion

In conclusion, this study, for the first time, reported the tet(X4) and tmexCD1-toprJ1 in a food-acquired R. ornithinolytica strain in China, evidenced by global R. ornithinolytica genomic analysis. Further, the identification of tet(X4) and tmexCD1-toprJ1 genes on separate conjugative plasmids in R. ornithinolytica indicates their expanding host range and independent horizontal transfer potential across bacterial species or genera which can pose a major threat and further dissemination in One Health components. Moreover, the convergence of complete yersiniabactin virulence apparatus in the current study strain suggests a significant threat to public health. We acknowledge that our study has some limitations such as small sample size, targeting one province, and selective tigecycline resistance isolation. Therefore, the present study recommends the need for large-scale continuous and vigilant genomic surveillance of bacterial strains carrying all critical plasmid-borne resistance genes in food supply chains to monitor and prevent their further dissemination in One Health settings.

CRediT authorship contribution statement

Muhammad Shoaib: Writing – original draft, Conceptualization, Writing – review & editing, Resources, Methodology, Investigation, Funding acquisition. Asim Munir: Writing – review & editing, Software, Formal analysis. Muhammad Fazal Hameed: Writing – review & editing, Software, Investigation. Shah Zeb: Data curation, Visualization, Writing – review & editing. Zhichao Li: Methodology, Writing – review & editing, Validation. Kai Peng: Writing – review & editing, Formal analysis, Validation. Nishant Shah: Writing – review & editing, Methodology, Investigation, Formal analysis. Patrick Butaye: Writing – review & editing, Investigation, Formal analysis. Zhiqiang Wang: Writing – review & editing, Resources, Funding acquisition. Ruichao Li: Project administration, Funding acquisition, Writing – review & editing.

Data availability statement

The complete assembled genome sequence data generated along with this study has been submitted in the NCBI under the BioProject ID: PRJNA1288727.

Funding

This work was supported by the Outstanding Youth Foundation of Jiangsu Province of China (BK20231524), National Natural Science Foundation of China (No. 32473095, 12411530085 and 32373061), National Natural Science Foundation of China for International Young Scientists (Grant No: 201012443) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Handling Editor: Professor Alejandro G.Marangoni

Footnotes

Appendix A

Supplementary data to this article can be found online at https://doi.org/10.1016/j.crfs.2026.101362.

Contributor Information

Zhiqiang Wang, Email: zqwang@yzu.edu.cn.

Ruichao Li, Email: rchl88@yzu.edu.cn.

Appendix A. Supplementary data

The following is the Supplementary data to this article:

Multimedia component 1
mmc1.docx (226.4KB, docx)

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Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

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Data Availability Statement

The complete assembled genome sequence data generated along with this study has been submitted in the NCBI under the BioProject ID: PRJNA1288727.

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