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. 2025 Sep 25;25:587. doi: 10.1186/s12866-025-04253-4

Faucicola mancuniensis isolated from human respiratory tract and its genomic characteristicstics

Tao Gu 1,#, Jian Zhou 1,2,#, Ying Liu 2, Qing Ma 2, Chunting Liu 2, Xiaoyu Wei 2, Yong Hu 1,✉, Shijun Li 1,2,✉
PMCID: PMC12465675  PMID: 40999358

Abstract

Background

Faucicola mancuniensis (F. mancuniensis), a Gram-negative bacterium within the Moraxellaceae family, has been isolated from the human respiratory tract. Genomic characteristics, antibiotic resistance profiles, and pathogenic potential of this rarely reported species remain largely unexplored. This study aimed to isolate F. mancuniensis from the human respiratory tract, characterize its genome, functional potential, and antimicrobial susceptibility, and assess its public health significance.

Methods

F. mancuniensis strain HZ006 was isolated from a human respiratory specimen in Guizhou, China. Taxonomic identification was confirmed using 16S rRNA gene sequencing. Morphology and physiological biochemistry were characterized. Whole-genome sequencing was performed using second- and third-generation technologies. Genomic functional annotation utilized GO, KEGG, VFDB, CARD, and four additional databases. Antimicrobial susceptibility against 29 antibiotics was determined phenotypically.

Results

16S rRNA sequencing confirmed strain HZ006 as F. mancuniensis (99.58% identity). HZ006 is an aerobic, Gram-negative coccobacillus (observed via electron microscopy), lacking flagella and spores. Optimal growth occurred at 18–37°C on blood agar, forming smooth, moist, circular colonies (~ 1.0 mm diameter). The complete genome is 2,740,046 bp with a G + C content of 40.38 mol%, encoding 2594 predicted genes. Annotation revealed 104 putative virulence factors (VFDB) and 27 antibiotic resistance genes (CARD), indicating potential pathogenicity and resistance mechanisms. Phenotypic testing, however, showed HZ006 was susceptible to all 29 antibiotics tested.

Conclusion

This study reports the first isolation and complete genome sequence of F. mancuniensis (HZ006) from a human specimen in China. Comprehensive genomic annotation and phenotypic characterization, including electron microscopy, provide foundational data on this species. While genomic analysis suggests potential pathogenicity and resistance mechanisms, the observed phenotypic susceptibility to all tested antibiotics informs potential treatment strategies. These findings significantly enhance our understanding of F. mancuniensis and its public health relevance.

Keywords: F. mancuniensis, 16S rRNA, Whole genome sequencing, Gene annotation, Electron microscope, Antibiotic sensitivity

Introduction

Faucicola mancuniensis (F. mancuniensis) is a Gram-negative, aerobic, non-motile coccus first isolated from human tonsils and taxonomically described in 2015 [1]. As the sole species within the genus Faucicola (family Moraxellaceae, class Gammaproteobacteria), it remains sparsely characterized since its discovery [2]. To date, genomic features, pathogenic potential, and antimicrobial resistance profiles of F. mancuniensis are largely unexplored, with no complete genome sequences available in public databases.

The strain examined in this study (HZ006) was isolated from throat swabs of adolescent close contacts (aged 13–14 years) of epidemic cerebrospinal meningitis (CSM) patients in Guizhou Province, China—historically a CSM-endemic region with seasonal incidence peaks [3]. Notably, these contacts were later confirmed negative for Neisseria meningitidis infection. Given that > 80% of CSM cases occur in children < 15 years and the absence of prior reports linking F. mancuniensis to meningococcal disease contacts, this isolate presents significant research value for understanding its ecological niche and clinical relevance [4, 5].

Although Humphreys et al. established the taxonomic identity of F. mancuniensis through 16S rRNA phylogeny and characterized basic physiological traits [1], comprehensive genomic and functional analyses are lacking. This study aims to: (i) provide the first complete genome sequence of F. mancuniensis using hybrid sequencing technologies, (ii) elucidate its genomic architecture and functional potential through multi-database annotation, (iii) characterize its morphological and biochemical features, and (iv) assess its antimicrobial susceptibility profile and virulence gene repertoire to evaluate public health significance.

Methods and materials

Ethics approval and informed consent

This study was conducted in accordance with China’s National Bacterial Disease Surveillance Program, involving continuous respiratory specimen collection from patients at sentinel hospitals. The Ethics Committee of Guizhou Provincial Center for Disease Control and Prevention reviewed the protocol and waived the requirement for formal ethical approval as the study constituted routine public health surveillance. All participants provided written informed consent prior to specimen collection and analysis.

Bacterial isolation and source

Strain HZ006 was isolated during an epidemic cerebrospinal meningitis (CSM) response in Guizhou Province. A total of 109 throat swabs were aseptically collected from meningitis cases, close contacts, and surrounding populations. Specimens were inoculated onto blood agar, Columbia blood agar, TSA (Soybean Casein Digest Agar), and BHI (Brain-heart infusion broth) (Guangdong Huankai Microbial Sci. & Tech. Co., Ltd, China) and incubated at 18°C, 28°C, and 37°C [2, 6]. Growth was monitored at 6 h, 12 h, 24 h, and 48 h. Distinct colonies were purified for subsequent analysis.

16S rRNA gene identification

Genomic DNA was extracted using heat lysis (95°C, 15 min). The near-full-length 16S rRNA gene was amplified with universal primers 27 F (5′-GAACCGCATGGTTCTTGGC-3′) and 1492R (5′-ACCCCACATCTCACGACACG-3′) using Premix Taq (Takara Bio, Shiga, Japan) under the following conditions: 94°C for 5 min; 35 cycles of 94°C for 15 s, 50°C for 5 s, 72°C for 1.5 min; final extension at 72°C for 5 min (25 µL reaction volume). Purified PCR products were subjected to bidirectional Sanger sequencing by Beijing Tianyi Huiyuan Co., Ltd. (China). Using the web-based BLASTn (https://blast.ncbi.nlm.nih.gov/Blast.cgi?PROGRAM=blastn&PAGETYPE=BlastSearch&LINKLOC=blasthome), the sequences were aligned with the targeted loci project database (16S rRNA) in the NCBI Reference Sequence Database (September 2024). Species identification required ≥ 98.7% similarity; values below this threshold suggested potential novel taxa [7–9]. Phylogenetic analysis was performed in MEGA11 [10] using neighbor-joining with 1000 bootstrap replicates.

Phenotypic characterization

A single colony of purified HZ006 strain was inoculated onto blood agar plates(Guangdong Huankai Microbial Sci. & Tech., Guangzhou, China) and cultured under optimal conditions (37°C for 24–48 h) until reaching the logarithmic growth phase. The bacterial morphology, including shape, color, texture, size, margin characteristics, and hemolytic activity, was observed. Gram staining was performed on strain HZ006 following standard protocols [6], with microscopic examination. Fresh single colonies obtained from purified HZ006 cultures were analyzed using the API 20NE strips (bioMérieux, France) test strips according to the manufacturer’s instructions. Logarithmic-phase HZ006 colonies were suspended and centrifuged to obtain bacterial pellets approximately the size of mung beans. After removing the supernatant, the pellets were fixed with Electron microscopy fixative at room temperature for 30 min in the dark. The fixed samples were then processed by Wuhan Servicebio Technology Co., Ltd for TEM and SEM analysis.

Whole-genome sequencing and assembly

High-molecular-weight DNA was extracted using the Magnetic Bacteria Genomic DNA Kit (RBJ1015) from Hangzhou Baiju Biotechnology Co., Ltd. Quantification was performed using Qubit 4.0 (Thermo Fisher Scientific, USA). Quality control was assessed by A260/A280 ratio > 1.8 and concentration > 40 ng/µL.

Qualified DNA samples, as confirmed by electrophoresis, were randomly fragmented to approximately 350 bp using a Covaris ultrasonicator. Using the NEBNext® Ultra™ DNA Library Prep Kit for Illumina (NEB, USA), the processed DNA fragments underwent end repair, A-tailing, adapter ligation, purification, and PCR amplification to complete library preparation. Following library construction, the library was initially quantified using Qubit 2.0 and diluted to 2 ng/µl. The insert fragment size of the library was then assessed using the Agilent 2100 Bioanalyzer. Upon confirmation that the insert size met the expected range, the library’s effective concentration was precisely quantified via Quantitative PCR (Q-PCR) to ensure its quality. Finally, sequencing was performed on the Illumina HiSeq.

The Multiple samples DNA Library Prep Kit for Ligation Sequencing (BK-AUX024) from Hangzhou BAIYITECH Co., Ltd. was used to construct libraries from quality-qualified DNA samples. Following steps of DNA repair and end-prep, Native barcode ligation, Adapter ligation, and clean-up, the library concentration (ng/µL) was precisely quantified using quantitative fluorometry (Qubit). Finally, the purified library was loaded onto an Oxford Nanopore Technologies (ONT) flow cell (FLO-MIN114). Sequencing was performed on a Nanopore GridION platform (Oxford Nanopore Technologies, UK) [11–13].

The final chromosomal assembly was generated using a hybrid approach with ABySS.Basecalling and de novo assembly were performed using MinKNOW v22.05 (Oxford Nanopore) [14]. The ABySS assembly was polished using Medaka v1.7.0 (https://github.com/nanoporetech/medaka) with default parameters, followed by secondary polishing with Pilon v1.22 incorporating Illumina sequencing data. Flye v2.9was solely employed for preliminary long-read assembly evaluation (not for final genome construction) [15–17].

Genome annotation and analysis

The whole genome assembly and annotation were performed using the Prokka v1.14.6 software [18], with default parameter settings adopted. To comprehensively characterize functional elements, predicted protein sequences were aligned against various functional databases using DIAMOND (e-value ≤ 1e-5). including NR(https://www.ncbi.nlm.nih.gov/refseq/about/nonredundantproteins/), COG (https://www.ncbi.nlm.nih.gov/research/cog/). Pathway analysis: KEGG (https://www.kegg.jp/). Functional classification: GO (http://geneontology.org/). Transporters: TCDB (http://www.tcdb.org/). Virulence factors and antibiotic resistance genes were identified using VFDB (https://www.mgc.ac.cn/VFs/) [19], CARD (https://card.mcmaster.ca/), and PHI-base (www.phi-base.org). For each sequence alignment, the top-scoring hit (default thresholds: identity ≥ 40%, coverage ≥ 40%) was selected for annotation. Circular genome visualization was generated with CGView Server (https://proksee.ca/).

Antimicrobial susceptibility testing

Susceptibility to 29 antibiotics was determined using broth microdilution (Thermo Fisher Sensititre™ CHN5FGNF and CHN6FGNF panels) according to CLSI guidelines. Briefly, HZ006 suspensions (0.5 McFarland standard) were diluted in CAMHB to ~ 5 × 10⁴ CFU/mL. Aliquots (50 µL) were dispensed into panels and incubated aerobically (35°C, 18–24 h). MIC values were interpreted using CLSI breakpoints (2024 edition) [20].

Results

Analysis of the 16S rRNA gene of the strain

Strain HZ006 was isolated and purified from 109 throat swab samples collected in Guizhou Province. DNA was extracted from strain HZ006, and the near-complete 16S rRNA gene sequence was amplified by PCR. The gene sequence was submitted to the NCBI database for BLAST comparison. BLAST results revealed that strain HZ006 showed the highest sequence similarity (99.58%) to the 16S rRNA gene of F. mancuniensis available in the database. This suggests that strain HZ006 and F. mancuniensis are conspecific. The 16S rRNA gene sequence of Aurantimonas litoralis LRZ36 was selected as the outgroup. Sequences from the NCBI database exhibiting similarity to the 16S rRNA gene sequence of strain HZ006 were selected for constructing the 16S rRNA gene phylogenetic tree. The results showed that the isolated strain HZ006 and F. mancuniensis clustered together on the same branch, indicating that the taxonomic position of strain HZ006 belongs to F. mancuniensis (Fig. 1).

Fig. 1.

Fig. 1

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The phylogenetic tree (Neighbour-joining) of the 16S rRNA gene sequence of strain HZ006

Physiological and biochemical characteristics

Experimental results showed that strain HZ006 was Gram-negative (Fig. 2B). The strain grew best on blood agar plates, with optimal growth achieved after 2-day incubation at 37°C. The growth temperature range was 18–37°C. After 24-hour culture on blood agar, circular colonies (≈ 1.0 mm diameter) with smooth, moist surfaces and entire margins were observed. No hemolysis was observed (Fig. 2A). HZ006 was observed under an electron microscope as a coccobacillus, without spores or flagella (Fig. 3A and B).

Fig. 2.

Fig. 2

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A Morphology of strain HZ006 B Gram staining of strain HZ006

Fig. 3.

Fig. 3

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A Morphology under Scanning Electron Microscope of strain HZ006 B Morphology under ransmission electron microscopic of strain HZ006

API 20NE results showed that strain HZ006 did not ferment glucose (GLU), arabinose (ARA), mannose (MNE), mannitol (MAN), N-acetylglucosamine (NAG), or maltose (MAL); and did not decompose potassium gluconate (GNT), capric acid (CAP), adipic acid (ADI), malic acid (MLT), sodium citrate (CIT), or phenylacetic acid (PAC); Oxidase test (OX), indole production (TRP), arginine dihydrolase (ADH), and urease (URE) tests were negative; It could reduce nitrate to nitrite but could not utilize ferric citrate.

Genome characteristics of strain HZ006

The complete genome of strain HZ006 is 2,740,046 bp in length with a G + C content of 40.38 mol%, containing 2594 predicted genes, including 12 rRNA, 47 tRNA and 2 ncRNA genes. The complete genome sequence of HZ006 has been deposited in NCBI under GenBank accession CP171363. The circular genome map comprehensively displays the genomic structure and features (Fig. 4). The innermost circle shows genomic coordinates, followed outward by ncRNA distribution, GC skew, GC content, and coding gene locations.

Fig. 4.

Fig. 4

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Whole genome circle of strain HZ006

Genomic function annotation of HZ006 gene

The GO database is divided into three categories: Cellular Component (CC), Molecular Function (MF), and Biological Process (BP). As shown in Fig. 5A, a total of 5,611 genes in strain HZ006 were annotated, with 860 genes annotated in CC, 1,895 in MF, and 2,856 in BP. The top three functional annotations in CC were cell anatomical entity (558), intracellular (207), and protein-containing complex (69); In MF, the top three were catalytic activity (855) reflecting robust energy metabolism. DNA binding (743), and transport activity (105); while in BP they were cellular process (890), metabolic process (872), and localization (291).

Fig. 5.

Fig. 5

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A GO analysis for the result of strain HZ006. B KEGG analysis for the result of strain HZ006. C NR analysis result of strain HZ006

The biological pathways in the KEGG database are classified into six major categories: including biological systems, genetic information processing, metabolism, cellular synthesis processes, environmental information processing, and human diseases. As shown in Fig. 5B, comparison of strain HZ006’s coding sequences with KEGG database resulted in annotation of 1,157 coding genes, among which metabolism-related genes were most abundant (623 genes), and organismal systems-related genes were least represented (41 genes). Human disease-related genes numbered 94, involving cancer, immune diseases, viral infections, parasitic infections, neurodegenerative diseases, bacterial infections, and drug resistance.

As shown in Fig. 5C of the NR database annotation results, 995 genes were annotated as Moraxella boevrei, accounting for the highest proportion, followed by 266 Moraxella osloensis (12.4%), and 196 Moraxella macacae (9.2%). The genus Moraxella had the most annotations.

As shown in Fig. 6A, the strain HZ006 has a total of 1,712 genes annotated into 24 COG functional categories. Among them, there are 201 gene functions related to translation, ribosome structure and biosynthesis; 130 genes related to amino acid transport and metabolism; 123 genes involved in cell wall/membrane/capsule biosynthesis; and 122 genes related to energy production and conversion.

Fig. 6.

Fig. 6

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A GOG analysis for the result of strain HZ006. B TCDB analysis for the result of strain HZ006. C PHI analysis result of strain HZ006

The annotation results of the TCDB database are shown in Fig. 6B. The strain HZ006 was annotated with a total of 192 genes. Among them, the primary active transport proteins were the most, with 93, followed by electrochemical potential-driven transport proteins with 49. TCDB transporters(93) suggest efficient nutrient scavenging-a key survival trait in nutrient-poor mucosal environments.

The PHI database annotation results are shown in Fig. 6C. Strain HZ006 contains 276 genes with PHI phenotype mutation types, including 1 gene for chemical resistance and 1 for chemical sensitivity, 2 factors determining phytotoxicity, 15 hypervirulence factors and 171 virulence attenuation factors.

As shown in Fig. 7, strain HZ006 was annotated with 27 resistance genes (identity > 40%) in the CARD database. HZ006 primarily exhibited four resistance mechanisms: antibiotic inactivation, antibiotic efflux, antibiotic target replacement, and antibiotic target alteration. These included 19 distinct resistance gene families, predominantly (RND) antibiotic efflux pumps, ribosomal RNA methyltransferases, and glycopeptide resistance gene clusters. The prediction of resistance genes in strain HZ006 suggests potential resistance to multiple antibiotics, including fluoroquinolones and cephalosporins.

Fig. 7.

Fig. 7

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CRAD analysis for the result of strain HZ006

By comparing the amino acid sequences of strain HZ006 with virulence factors in the VFDB database (Table 1). A total of 104 virulence genes were annotated in the VFDB database. distributed across 61 gene entries. This study identified virulence genes including lipB, hisF, pilU, lic2A, lgtF, bfmR, and wzm in the HZ006 genome. These genes encode products potentially harmful to host cells, including capsular polysaccharide modification proteins, N-acetylglucosamine deacetylase, capsular polysaccharide export ABC transporter transmembrane proteins, twitching motility proteins, imidazole glycerol phosphate synthase, and 1,4-glucosyltransferase.

Table 1.

Virulence gene analysis of VFDB

Virulence gene type VFDB Family VFDB factors VFDB gene
Regulation of virulence-associated genes Regulation Fur fur
BfmRS bfmR
Offensive virulence factors Toxin LOS lpxH 、 kdsA 、 galU 、 orfM 、 yhxB/manB 、 lsgF 、 lgtF 、 lic2A
Secretion system Capsule ABZJ_00085 、 lipB 、 ABZJ_00095 、 ABK1_0086 、 ACICU_00076 、 ACICU_00078 、 ACICU_00080 、 ABK1_0095 、 ABTW07_0090 、 cpsB
LPS kdtB 、 lpxC 、 hisF 、 acpXL 、 lpxB 、 lpxD 、 fabZ 、 lpxA 、 lpxL
PbpG pbpG
Adherence Hsp60 htpB
MOMP CT396
Tap type IV pili tapQ
Type IV pili pilX 、 pilE 、 pilG 、 pilU 、 pilT2
Defensive virulence factors Antiphagocytosis Capsule I wzt2 、 wzm 、 wcbD 、 wcbC
Stress protein ClpP clpP
SodB sodB
ClpE clpE
Nonspecific virulence factor Iron uptake system CcmC ccmC
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Antibiotic resistance test results

The antimicrobial susceptibility and minimum inhibitory concentration (MIC) test results of strain HZ006 are shown in Table 2. The strain was susceptible to ampicillin (AMP), ampicillin/sulbactam (AMS), cefoxitin (CFX), cefuroxime (CXM), cefepime (CPM), ceftazidime (CAZ), ceftazidime/clavulanic acid (CAZ/C), cefotaxime (CTX), cefotaxime/clavulanic acid (CTX/C), chloramphenicol (CHL), colistin (CT), ciprofloxacin (CIP), nalidixic acid (NAL), cefazolin (CFZ), ceftiofur (CEF), ceftazidime/avibactam (CZA), imipenem (IPM), ertapenem (ETP), amoxicillin/clavulanic acid (AMC), gentamicin (GEN), amikacin (AMK), streptomycin (STR), tetracycline (TET), meropenem (MEM), tigecycline (TIG), polymyxin B (PB), azithromycin (AZM), trimethoprim/sulfamethoxazole (SXT), and florfenicol (FFC).

Table 2.

Concentration of minimum growth inhibitory and antimicrobial susceptibility of strain HZ006 (Note S-Sensitive, I-Intermediary, R-Resistant)

Antibiotics MIC Interpretation Antibiotics MIC Interpretation
AMP <=1 S CEF <=0.5 S
AMS <=1 S CZA <=0.5 S
CFX <=1 S IPM <=0.25 S
CXM <=1 S ETP <=0.25 S
CPM <=1 S AMC <=1 S
CAZ <=0.5 S GEN <=1 S
CAZ/C <=0.25 S AMK <=4 S
CTX <=0.12 S STR <=4 S
CTX/C <=0.06 S TET <=1 S
CHL <=2 S MEM <=0.12 S
CT <=0.25 S TIG <=0.25 S
CIP <=0.015 S PB <=0.25 S
NAL <=2 S AZM <=2 S
CFZ <=1 S SXT <=0.25 S
CEF <=0.5 S FFC <=1 S
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Discussion

This study isolated and cultured F. mancuniensis (HZ006) from 109 throat swab samples. The samples were collected from the respiratory tract of close contacts of meningitis patients. This isolation site is consistent with where Gavin J. Humphreys first discovered F. mancuniensis. Gavin J. Humphreys et al. only performed biochemical characterization including fatty acids, respiratory quinones, oxidase, and catalase tests. This study represents the first identification of F. mancuniensis in China. We conducted whole genome sequencing and supplemented with relevant biochemical and antibiotic resistance phenotypic tests.

Currently, comparative genomics approaches combining whole genome sequencing with database alignment analysis are widely used in studies exploring bacterial gene functions [21]. This approach not only provides in-depth understanding of bacterial genetic information, but also enables identification of virulence and antimicrobial resistance genes through comparison with known functional gene genomes. In this study, traditional isolation and culture methods were employed to cultivate the samples for meningococcal surveillance from the population in Guizhou Province, China. The isolated and purified strains were identified using the 16S rRNA gene analysis technique to determine their identities. Phylogenetic analysis of 16S rRNA gene sequences confirmed the isolation of F. mancuniensis (HZ006) from pharyngeal swab specimens. Strain HZ006 showed closest homology to F. mancuniensis (GenBank accession KC688888) previously reported by Gavin J. Humphreys [1].

NR database species annotation showed that the genus Moraxella was the most abundant, which was consistent with phylogenetic tree analysis indicating the closest genetic relationship between Moraxella and F. mancuniensis. In COG, GO, and KEGG databases, we identified multiple genes and metabolic pathways related to metabolism, energy conversion, and substance transport, suggesting vigorous metabolic activity in strain HZ006. KEGG annotation revealed 94 human disease-related pathways, including those associated with neurodegenerative diseases, infectious diseases, and drug resistance, suggesting potential health risks posed by F. mancuniensis [22, 23]. TCDB annotation identified 192 genes in strain HZ006, predominantly 93 primary active transporters, followed by electrochemical potential-driven transporters, consistent with the Membrane transport pathway in KEGG’s environmental information processing. These transporters play crucial roles in biosynthesis and secretion, and are also involved in energy-releasing processes like photon absorption, substrate decarboxylation, and methyl transfer [24]. PHI analysis identified 276 phenotype mutation-associated genes in HZ006, including chemical resistance, phytotoxicity determinants, and hypervirulence factors, suggesting F. mancuniensis may possess pathogenic potential against human health [25].

The genome sequence of F. mancuniensis was analyzed against the VFDB database, annotating 104 virulence genes. The genome contains several virulence genes affecting host cells, including lpxC, lpxD, wzm, and lgtF. lpxC and lpxD have been demonstrated to trigger host inflammatory responses [26–28], wzm serves as a crucial virulence determinant [29, 30], it reduces bacterial phagocytosis by host cells through inhibiting C3b complement deposition on the bacterial surface [31, 32], lgtF contributes to immune evasion by binding to asialoglycoprotein receptors on host cells [33, 34]. These functional annotations and pathway predictions suggest F. mancuniensis may pose risks to humans. It also provides a direction for further in-depth research.

The biochemical test results were generally consistent with those reported by Gavin J. Humphreys [1]. Except that the oxidase test result was inconsistent with previous reports. Additionally, this study demonstrated that the strain cannot metabolize potassium gluconate (GNT), capric acid (CAP), adipic acid (ADI), malic acid (MLT), sodium citrate (CIT), or phenylacetic acid (PAC). Antimicrobial susceptibility testing revealed that strain HZ006 was sensitive to all 29 tested antibiotics including cefoxitin (CFX), ertapenem (ETP), and gentamicin (GEN). This provides guidance for future antibiotic treatment of human infections caused by this bacterium. Furthermore, the electron microscope images show that F. mancuniensis does not possess any flagella or cilia. This is significant for understanding its motility, adhesion, adaptability, and pathogenicity in humans [35, 36].

Conclusion

This study first obtained and published the complete genome sequence data of F. mancuniensis, and analyzed its morphological, physiological and biochemical characteristics. Genetic predictions were made regarding its pathogenicity and signaling pathways. Supplementing the biochemical experimental results of this bacterium. and describing its ultrastructural characteristics under electron microscopy. providing a foundation for subsequent research on this bacterium. While virulence genes were annotated, their expression and clinical relevance require functional validation in host-relevant models. Future work should validate pathogenic mechanisms under simulated host environmental conditions, clarify its role in the recovery capacity of respiratory tract microbial communities.

Acknowledgements

No.

Authors’ contributions

TG and JZ designed the study, conducted the study trial, and drafted the manuscript, YL and QM assisted in analyzing the data, XYW and CTL provided technical support, YH and SJL is the funding source, supervisor, and revised the manuscript.

Funding

1.This study was supported by ‘Project for Public Health Talent Cultivation of China. Grant No. Guo Jikong Zong Ren Han [2024]122’.

2.‘ZDSYS[2023]004(Key Laboratory of Microbiol and Infectious Disease prevention &Control in Guizhou Province. Grant No. Qiankehe Platform Talent-ZDSYS[2023]004)’.

3.‘The local science foundation of Guizhou Province guided by the Central Committee of China, No.Qiankehe [2025] 024’.

Data availability

The complete data set was submitted to the National Biotechnology Information Center (NCBI) database（Accession number：CP171363.1）. Data has been uploaded to the website https://www.ncbi.nlm.nih.gov/bioproject/PRJNA1172360. We have uploaded the raw sequencing data to the SRA database of NCBI. The accession numbers are: SRR34576495 (ONT) and SRR34576496 (Illumina). BioSample: SAMN50011251 (https://dataview.ncbi.nlm.nih.gov/object/59017646).

Declarations

Ethics approval and consent to participate

This study was conducted in accordance with China’s National Bacterial Disease Surveillance Program, involving continuous respiratory specimen collection from patients at sentinel hospitals. The Ethics Committee of Guizhou Provincial Center for Disease Control and Prevention reviewed the protocol and waived the requirement for formal ethical approval as the study constituted routine public health surveillance. All participants provided written informed consent prior to specimen collection and analysis. This research complies with the requirements of the Helsinki Declaration.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Footnotes

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Tao Gu and Jian Zhou joint first authors who contributed equally to this work.

Contributor Information

Yong Hu, Email: huyong1979@gmc.edu.cn.

Shijun Li, Email: zjumedjun@163.com.

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Associated Data

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

Data Availability Statement

The complete data set was submitted to the National Biotechnology Information Center (NCBI) database（Accession number：CP171363.1）. Data has been uploaded to the website https://www.ncbi.nlm.nih.gov/bioproject/PRJNA1172360. We have uploaded the raw sequencing data to the SRA database of NCBI. The accession numbers are: SRR34576495 (ONT) and SRR34576496 (Illumina). BioSample: SAMN50011251 (https://dataview.ncbi.nlm.nih.gov/object/59017646).

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