The transmission pathways and risks of COVID-19-associated pulmonary aspergillosis (CAPA) remain unclear. This study investigated the genetic relationships of isolates from patients with and without COVID-19 and environmental air samples to suggest possible transmission patterns. We conducted a prospective study from March 2020 to December 2022, collecting clinical and environmental isolates from a tertiary hospital. Isolates from patients with and without COVID-19 were compared with those from air samples at four hospital locations. The genetic analysis included internal transcribed spacer and sequencing, with azole resistance assessed via gene analysis. Multiple locus variable-number tandem repeat analysis was performed to elucidate genetic relationships. A total of 155 isolates (19 from COVID-19 patients, 104 from non-COVID-19 patients, and 32 from environmental samples) were identified and genotyped, revealing 131 sequence types (Simpson Diversity Index 0.9972). Four CAPA clinical strains genetically related to environmental strains were isolated from the COVID-19 intensive care unit (ICU), while two CAPA clinical strains sharing multiple locus variable-number tandem repeat sequence types and azole-resistant mutations were isolated in the same COVID-19 ICU 4 months apart. All but one of these strains were isolated from patients requiring mechanical ventilation. The observed genetic similarities between strains from critically ill patients with COVID-19 and those from the environment, as well as within the same ICU, raise the possibility of nosocomial acquisition via contaminated air or environmental sources. These findings highlight the risks of CAPA associated with negative pressure rooms and the need for enhanced environmental infection control measures.IMPORTANCEThis study reveals genetic links between in patients with COVID-19 and environmental sources, suggesting nosocomial transmission and urging a reevaluation of universal negative pressure isolation practices in hospitals, especially for critically ill patients.

(MS) is an important pathogen in the poultry industry and has caused significant economic losses. Worldwide, the use of live attenuated vaccine for the MS-H strain has increased to prevent MS infection. However, there is no test available to discriminate the MS-H vaccine strains from the MS strains that are causing field infection. In this study, a TaqMan-MGB real-time PCR method (qPCR) was established, validated, and evaluated to discriminate between MS-H-live vaccine and field strains based on nucleotide differences in the gene. The validation was performed for sensitivity and reproducibility by constructing recombinant plasmids. The limits of detection were 1.07 × 10 copies/µL for the MS-H and 1.95 × 10 copies/µL for field strains, respectively. The intra- and inter-assay results were less than 2.5% based on the reproducibility test. No cross-amplification signals from other common chicken pathogens were detected. Thus, our data indicated that this qPCR is sensitive, specific, and reproducible. In addition, 709 chicken clinical samples were used to evaluate this qPCR test. The results showed that positive signals could be detected from the chicken choanal cleft swabs and are 100% in concordance with the PCR sequencing method. To the best of our knowledge, we found for the first time that both L- and C-type field MS were present in flocks immunized against the MS-H vaccine strain during the validation process. In addition, this is the first report of a field strain of C-type in China.IMPORTANCE (MS) is an important pathogen in the poultry industry and has caused significant economic losses. Worldwide, an increasing number of farms are using the live attenuated vaccine MS-H strain to prevent MS infections. In order to monitor vaccinated and naturally infected flocks and to continue the MS control and eradication program, a differentiation of infected from vaccinated animals (DIVA) test for MS is urgently needed. We developed a TaqMan-MGB real-time qPCR (qPCR) method with a pair of primers and two competitive TaqMan-MGB probes. We performed an evaluation that can discriminate between the MS-H-live vaccine and field MS strains based on nucleotide differences in the gene. It has great sensitivity and reproducibility, and greater specificity than other methods which were established by SNP sites of the gene and gene.

Dehong virus (DEHV) is an emerging filamentous virus of considerable interest. However, research involving DEHV remains limited, and no suitable models exist to investigate its pathogenicity or transmission. In this study, we developed an neutralization assay to detect DEHV-neutralizing antibodies, as well as an bioluminescent imaging mouse model based on a pseudovirus system. Our results confirmed that DEHV utilizes the Niemann-Pick disease, type C1 (NPC1) receptor for cellular entry. Additionally, the neutralization assay demonstrated that DEHV antiserum does not exhibit neutralizing activity against Mengla or Marburg viruses. This pseudovirus-based system provides a valuable platform for studying DEHV biology and evaluating therapeutic interventions.IMPORTANCEBats serve as natural reservoirs for diverse filoviruses across Africa, Europe, and East Asia; numerous strains circulate within these populations. Recently, Chinese researchers identified Dehong virus (DEHV), a novel filovirus carried by bats in China. However, the mechanisms underlying the pathogenicity and transmission of DEHV remain poorly understood. Similar to Ebola virus and Marburg virus (MARV), DEHV uses the Niemann-Pick disease, type C1 (NPC1) receptor for host cell invasion. In this study, we utilized a well-established in neutralization assay to confirm that DEHV antiserum lacks neutralizing activity against Mengla and MARV pseudoviruses. Furthermore, we developed an innovative in bioluminescent imaging mouse model using DEHV pseudovirus, which offers a visually intuitive and efficient platform for evaluating antiviral therapies and vaccine candidates. This model has considerable potential for advancing research into DEHV pathogenesis and treatment strategies.

African swine fever (ASF) is a highly infectious and lethal swine disease, leading to enormous losses in the pig industry. K205R, a non-structural protein of ASF virus (ASFV), is abundantly expressed at the early stages of viral infection and induces a strong immune response. In our previous study, five strains of K205R-specific nanobodies (Nbs) were screened through phage display technology, among which Nb1, Nb14, Nb35, and Nb82 exhibited good affinity. In the present study, the above four Nbs were successfully expressed in HEK293T cells and exhibited strong reactivity. Four Nbs recognized linear B-cell epitopes of K205R in both prokaryotic and eukaryotic expression systems. Besides, four Nbs specifically reacted with the K205R protein of ASFV-infected cells. Two epitopes MVEPR and RTQF were further identified, with highly conserved in different ASFV strains, and could interact with inactivated ASFV-positive sera, indicating that the two epitopes were natural linear B-cell epitopes. Moreover, structural analysis indicated that both epitopes were exposed on the surface of the K205R molecule. Notably, the identified epitope RTQF was first reported. Overall, these findings provide valuable insights for K205R as an effective diagnostic tool and vaccine development.IMPORTANCEAfrican swine fever (ASF) is the number one killer affecting the pig industry, and there are no effective strategies for prevention. The ASFV K205R protein is prominently expressed in the early stages of viral infection, triggering a robust immune response. The full understanding of K205R protein epitopes provides a theoretical basis for the development of vaccine-candidate proteins. Nanobodies exhibit superior capability in detecting concealed epitopes of antigens compared with traditional antibodies. Here, we identify two epitopes MVEPR and RTQF based on nanobodies as a tool. Notably, the epitopeRTQF is being reported for the first time. These epitopes are highly conserved in different ASFV strains and represent natural linear B-cell epitopes. This study opens up nanobodies as a new tool for the identification of epitopes and also provides a direct material basis for the development of ASFV vaccines.

Although the high-altitude limit for microbial survival in the Earth-atmosphere system has remained a scientific curiosity and topic of study, the ecological significance of long-distance microbial dispersal in the atmosphere has been perceived to have marginal relevance. Here, we report the characterization of novel plant pathogenic species of that were isolated from samples collected at altitudes ranging from 1.5 to 29 km above sea level. Whole genome-based phylogenies of three strains, paired with plant challenge assays, indicate that each is a previously unrecognized species and causes disease on beans comparable to . Isolates from the stratosphere (strain L6-1) and agricultural millet (G77) were identified to be the same species and designated as sp. nov. displays high levels of tolerance to desiccation and UV radiation, which are stresses that increase in intensity with altitude. Back trajectory air mass analysis implied that the phytopathogens may have had an intercontinental source, but regional origins in the continental US cannot be excluded. The environmentally robust phytopathogens we have documented in the upper atmosphere provide new perspective on the role that high-altitude transport may play in microbial dispersal, gene flow, and the epidemiology of aerially dispersed plant disease.

a diverse group of filamentous bacteria found predominantly in soil, play a crucial role in nutrient cycling and produce many valuable secondary metabolites for the pharmaceutical industry. In this pilot study, we collected 19 soil samples from 14 provinces in China to preliminarily investigate the biodiversity and genetic structure of in soils of China from different dimensions, using recently developed cost-efficient amplicon and whole-genome library preparation methods. Amplicon analysis showed that Actinobacteria were among the most abundant bacteria, with 0.3% of amplicon sequence variants (ASVs) belonging to . Meanwhile, we successfully isolated 136 natural strains and assembled their genomes, including 26 previously unreported species, underscoring the need for further exploration of soil in China. Population genetics analysis revealed that homologous recombination may primarily drive the extensive genetic diversity observed in , as well as a complex population structure. Complementing this, pan-genome analysis shed light on gene diversity within and led to the discovery of rare genes, further emphasizing the vast genetic diversity of this genus. Additionally, multiple metabolic gene clusters were found in these strains, as well as some potentially unique or uncommon ones were found. These findings not only highlight the biological and metabolic diversity of but also provide a technical framework for future studies on the global biodiversity and evolution of this genus.

Current treatments for chronic hepatitis B (CHB) virus involve nucleos(t)ide analogs and pegylated interferon-alpha (PEG-IFNα). This study compares the efficacy and safety of PEG-IFNα monotherapy with its combinations with entecavir (ETV) and tenofovir disoproxil fumarate (TDF) in managing CHB. We included 147 treatment-naïve patients divided into three groups: Group A (PEG-IFNα-2b with ETV), Group B (PEG-IFNα-2b with TDF), and Group C (PEG-IFNα-2b monotherapy). Evaluations occurred every 12 weeks up to 48 weeks. The Kaplan-Meier method showed no significant differences in cumulative HBsAg loss, but HBV DNA clearance rates were higher in the TDF group than in the ETV group ( = 0.01). Higher incidences of elevated alanine aminotransferase (ALT), aspartate aminotransferase, and thrombocytopenia were observed in the TDF group compared to other groups. After propensity score matching, the TDF group had a higher undetectable HBV DNA rate than the IFN group, but no significant differences in HBsAg clearance rates. Both TDF and ETV groups achieved more significant HBsAg reductions from baseline to week 48 than the IFN group ( < 0.05). ETV showed a lower HBeAg clearance rate (30.00% vs 87.50%, < 0.05) but higher ALT normalization (76.92% vs 45.45%, < 0.05). In the TDF group, patients with lower baseline HBsAg levels, high ALT levels, and lower aspartate aminotransferase-to-platelet ratio index (APRI) scores were more likely to achieve HBsAg loss. These findings suggest that TDF and ETV are effective for viral suppression, with TDF showing superior HBV DNA clearance but more adverse events.

Patients afflicted with Wilson's disease (WD) may encounter hepatic and extraneous manifestations due to the progressive accumulation of copper in the liver and other subsequent organs. Copper-chelating agents, such as D-penicillamine (DPA), are commonly utilized in the medical treatment of copper overload in WD. Manipulating the composition of gut microbiota appropriately can enhance drug efficacy and safety. This study aims to investigate how targeted intervention on gut microbiota influences the effectiveness of copper removal in a WD model during DPA treatment. First, following a 4-week treatment of DPA, the liver copper concentration and gut microbial composition were assessed in the WD mice model to identify potential candidates for targeted regulation of gut microbiota. Second, after 8 weeks of manipulating the gut microbiota during DPA treatment, various parameters including blood liver function indicators, tissue copper load, hepatic histopathological features, and gut microbiota were investigated in WD mice. The findings demonstrated that the presence of significantly enhances the efficacy of DPA, leading to a more efficient elimination of copper from tissues and a greater improvement in liver injury, liver dysfunction, and gut dysbiosis. In contrast, has an antagonistic effect. The results of gene function prediction analysis indicated that the altered gut microbial function by DPA and Akk is primarily linked to energy generation/utilization, amino acid, fatty acid, lipid, and nucleic acid metabolisms. To summarize, this study provides experimental evidence for the potential application of targeted regulation of gut microbiota in the adjunctive therapy of copper dysregulation disease.IMPORTANCECopper is an essential element in virtually all living organisms. Wilson's disease (WD) is a representative disorder caused by the disruption of copper homeostasis. Oral-chelating agents are the first-line treatment for copper-overloaded diseases, with D-penicillamine (DPA) being the prototypical drug. However, the efficacy and adverse effects of DPA remain challenging in its use for WD treatment. In our study, the supplementation of (Akk), a key gut microbe, along with DPA was demonstrated to enhance copper removal, ameliorate liver injury and dysfunction, and restore gut dysbiosis in a mouse model of WD. These findings highlight the significant potential applications of targeted modulation of gut microbiota as "pharmacomicrobiomics" in adjunctive therapy for disorders involving copper dysregulation.

We investigated the presence of viral DNA and RNA in cutaneous squamous cell carcinoma (cSCC) tumor and normal tissues from nine individuals with a history of hematopoietic stem cell transplantation (HCT). Microbiome quantification through DNA and RNA sequencing (RNA-seq) revealed the presence of 18 viruses in both tumor and normal tissues. DNA sequencing (DNA-seq) identified , , , , , , and others. RNA-seq revealed additional viruses such , , , , , , , and others. Notably, DNA-seq indicated that tumor samples exhibited low levels of in three out of nine subjects and elevated levels of in one subject, while normal samples frequently contained and occasionally . A comparative analysis using both DNA- and RNA-seq captured three common viruses: , , and . These findings were corroborated by an independent data set, supporting the reliability of the viral detection methods utilized. The study provides insights into the viral landscape in post-HCT patients, emphasizing the need for comprehensive viral monitoring in this vulnerable population.

The order consists of over 300 species of segmented, negative-strand RNA viruses. These viruses have a tri-segmented genome (S, M, and L segments) that encodes the nucleocapsid protein (N protein), glycoprotein precursor, and RNA-dependent RNA polymerase (RdRp), respectively. The RdRp is a large protein (~420 kDa) responsible for synthesizing viral mRNA and replicating the viral genome. Bunyaviruses initiate transcription of viral mRNA through a unique cap-snatching mechanism. During this process, the N-terminal endonuclease domain of the RdRp cleaves host cell mRNA at the 5' terminus and uses the resulting capped mRNA fragment as a primer. This endonuclease domain exhibits a highly conserved structural architecture and is essential for establishing viral infection in host cells. Therefore, the N-terminal endonuclease domain represents a promising target for therapeutic intervention against Bunyaviruses, particularly at the early stages of the virus replication cycle. In this study, we developed a highly sensitive fluorescence resonance energy transfer-based assay to quantitatively examine the activity of the bacterially expressed and purified endonuclease domain of hantavirus RdRp. A 20-nucleotide synthetic RNA, labeled with a 6-FAM fluorophore at the 5' end and an Iowa Black quencher at the 3' end, generated a significant dequenched fluorescence signal upon cleavage by the purified endonuclease domain. Kinetic analysis revealed a half-life () of approximately 3 min for the reaction, achieving a signal-to-background ratio of approximately 31.

Porcine circovirus type 2 (PCV2) serves as the key pathogen linked to porcine circovirus-associated disease, representing a considerable risk to the worldwide swine industry. A blocking enzyme-linked immunosorbent assay (ELISA) was established to identify antibodies specifically targeting PCV2, employing recombinant Cap protein as the antigen and a monoclonal antibody against PCV2 Cap protein as the detector antibody. Utilizing receiver operating characteristic curve analysis, a cutoff value of 33.8% was determined to distinguish between positive and negative serum samples. The sensitivity and specificity of this blocking ELISA method were reported at 94.7% and 96.1%, respectively. Notably, this approach exclusively identified antibodies for PCV2, showing no cross-reactivity with antibodies related to African swine fever virus (ASFV), Porcine epidemic diarrhea virus (PEDV), Porcine pseudorabies virus (PRV), and porcine reproductive and respiratory syndrome virus. Both intra-assay and inter-assay coefficients of variation were less than 10%. In a comparison involving 402 porcine serum samples, the agreement rate with a commercial indirect ELISA kit reached 98.76%, with a kappa value of 0.888, reflecting high concordance between the two testing methods. This study showcases the blocking ELISA method as an efficient and standardized approach for serological monitoring of PCV2 in swine populations and for assessing seroconversion in vaccinated pigs.

Focal duodenal necrosis (FDN) is an intestinal disease causing significant economic losses in the table-egg industry due to reduced egg production in laying hens. Its etiology and pathogenesis remain poorly understood. Between 2021 and 2023, 111 isolates were collected from FDN lesions and screened for the presence of virulence genes using PCR panels. Five strains-FDN-4, FDN-9, FDN-11, FDN-24, and FDN-50-were selected for whole-genome sequencing due to their high virulence gene content. Core-genome analyses found that the five FDN belong to different phylogroups and strain types (ST), but they all share multiple complete operons involved in key pathogenic functions, including host cell adhesion and invasion, iron acquisition, motility, biofilm formation, and acid resistance. Comparative genomic analyses identified FDN-4 as the most genetically distinct strain, closely resembling EC958, an O25b:H4 ST131 uropathogenic (UPEC) commonly associated with extended-spectrum beta-lactamase production. FDN-4 and EC958 share unique chromosomal virulence genes absent in the other FDN strains, all located within genomic islands. This study provides the first complete genomic characterization of isolated from FDN lesions and highlights FDN-4 as a genetically distinct strain with similarities to O25b:H4 ST131 UPEC.IMPORTANCEThis study presents the first complete genomic characterization of isolated from focal duodenal necrosis (FDN) lesions. Notably, FDN-4 is the first strain from a poultry disease (FDN) to show significant similarity to O25b:H4 ST131 strains, commonly classified as uropathogenic and often associated with extended-spectrum beta-lactamase production. However, caution is warranted when attributing direct transmission routes between poultry and humans.

The etiological spectrum of microbial keratitis exhibits significant regional variability. However, existing domestic and international resistance monitoring systems do not adequately address the specific needs of clinical practice. We analyzed a cohort of 3,691 patients diagnosed with microbial keratitis in the southern region of Zhejiang Province, China between 2016 and 2023. The patients' ages ranged from 2 to 93 years (979 men and 403 women). The microbial culture positivity rate is relatively low (38.72%). Trauma and foreign body entry emerged as significant risk factors. Mixed infections accounting for 4.85% posed challenges for diagnosis and treatment. Filamentous fungi, predominantly spp., dominated the microbial landscape. Prominent bacterial pathogens included , , , and was an important pathogen affecting the cornea in this region. The observed resistance patterns emphasized the urgency for alternative therapeutic strategies targeting resistant gram-positive (e.g., erythromycin, penicillin, oxacillin) and -negative (e.g., trimethoprim-sulfamethoxazole, tetracycline) bacteria as well as refractory fungi, such as (voriconazole-resistant) and (resistant to itraconazole and amphotericin B). Resistance to ceftazidime, meropenem, and erythromycin exhibited a slight upward trend, diverging from the overall bacterial resistance trend observed in China. High isolation rates of methicillin-resistant and macrolide-resistant underscored the need for enhanced infection control measures and targeted interventions against these resistant pathogens. This study elucidated the evolving patterns of antibiotic resistance among ocular isolates in the region, providing a critical foundation for the effective application of antimicrobial therapies in clinical practice.IMPORTANCECurrently, our region does not possess extensive monitoring data regarding antibiotic resistance trends in ocular isolates, especially those derived from corneal infections. This study addresses a critical component of ophthalmic microbiology by analyzing long-term data to identify trends in etiological agents and their clinical implications. It aimed to fill the void in epidemiological data on ocular isolates within our region, providing scientific insights essential for the comprehensive monitoring of ocular microbial drug resistance.

Predicting vancomycin exposure is essential for optimizing dosing regimens in sepsis patients. While population pharmacokinetic (PPK) models are commonly used, their performance is limited. Machine learning (ML) models offer advantages over PPK models, but it remains unclear which model-PPK, Bayesian, ML, or hybrid PPK-ML-is best for predicting vancomycin exposure across different clinical scenarios in sepsis patients. This study compares the performance of these models in predicting the 24 hour area under the blood concentration curve (AUC) to support precision dosing in sepsis care. Data from sepsis patients treated with intravenous vancomycin were sourced from the MIMIC-IV database. The data set was split into training and testing sets, and four models-PPK, Bayesian, ML, and hybrid-were developed. In the testing set, AUC was predicted using all models, and performance was evaluated using mean absolute error, mean squared error, root mean squared error, mean absolute percentage error (MAPE), and R². A total of 4,059 patients were included. In the absence of vancomycin concentration data, the hybrid model outperformed both PPK and Bayesian models, with MAPE improvements of 58% and 17%, respectively. When vancomycin concentration data were available, the Bayesian model demonstrated the best performance (MAPE: 13.37% vs 68.17%, 34.17%, and 28.52% for PPK, Random Forest, and hybrid models). The hybrid model is recommended to predict AUC when concentration data were unavailable, while the Bayesian model should be used when concentrations were available, offering robust strategies for precise vancomycin dosing in sepsis patients.

is an opportunistic pathogen that can cause sinus infections and pneumonia in cystic fibrosis (CF) patients. Bacteriophage therapy is being investigated as a treatment for antibiotic-resistant infections. Although virulent bacteriophages have shown promise in treating infections, the development of bacteriophage-insensitive mutants (BIMs) in the presence of bacteriophages has been described. The aim of this study was to examine the genetic changes associated with the BIM phenotype. Biofilms of three genetically distinct strains, including PAO1 (ATCC 15692), and two clinical respiratory isolates (one CF and one non-CF) were grown for 7 days and treated with either a cocktail of four bacteriophages or a vehicle control for 7 consecutive days. BIMs isolated from the biofilms were detected by streak assays, and resistance to the phage cocktail was confirmed using spot test assays. Comparison of whole genome sequencing between the recovered BIMs and their respective vehicle control-treated phage-sensitive isolates revealed structural variants in two strains, and several small variants in all three strains. These variations involved a TonB-dependent outer membrane receptor in one strain, and mutations in lipopolysaccharide synthesis genes in two strains. Prophage deletion and induction were also noted in two strains, as well as mutations in several genes associated with virulence factors. Mutations in genes involved in susceptibility to conventional antibiotics were also identified in BIMs, with both decreased and increased antibiotic sensitivity to various antibiotics being observed. These findings may have implications for future applications of lytic phage therapy.IMPORTANCELytic bacteriophages are viruses that infect and kill bacteria and can be used to treat difficult-to-treat bacterial infections, including biofilm-associated infections and multidrug-resistant bacteria. is a bacterium that can cause life-threatening infections. Lytic bacteriophage therapy has been trialed in the treatment of infections; however, sometimes bacteria develop resistance to the bacteriophages. This study sheds light on the genetic mechanisms of such resistance, and how this might be harnessed to restore the sensitivity of multidrug-resistant to conventional antibiotics.

Subclinical tuberculosis (TB) challenges public health interventions. This retrospective multicenter study aimed to evaluate the diagnostic efficacy of smear microscopy, mycobacterial culture, and GeneXpert MTB/RIF assay (Xpert) for subclinical TB and identify its proportion and risk factors in China. Among 560 participants diagnosed with TB, 309 (55.18%) patients had active TB, and 251 (44.82%) had subclinical TB. For subclinical TB diagnosis, smear microscopy, mycobacterial culture, and Xpert had detection rates of 11.5%, 40.6%, and 40.6%, respectively, which were significantly lower than those for active TB (25.2%, 59.2%, and 63.7% respectively). Conversely, interferon-γ release assays (IGRA) showed similar detection rates for both subclinical (85.7%) and active TB (84.1%). Age groups of 40-60 years and under 40 years, as well as employed individuals had higher risks of subclinical TB. To conclude, about 45% of TB patients were subclinical in our study. Microbiologic tests had lower diagnostic sensitivities for subclinical TB than for active TB, while IGRA positivity rates were comparable between asymptomatic and symptomatic TB patients, highlighting the potential of antigen-specific immune response in the diagnosis of these asymptomatic individuals. Further clinical trials are warranted to investigate the diagnostic flow chart in individuals with subclinical TB.

Mice are colonized by diverse gut fungi, known as the mycobiota, which have received much less attention than bacterial microbiota. Here, we studied the diversities and structures of cecal fungal communities in wild (, and ) vs laboratory C57BL/6J mice to disentangle the contributions of gut fungi to the adaptation of mice to genetic diversity. Using ITS1 gene sequencing, we obtained 2,912 amplicon sequence variants (ASVs) and characterized the composition and diversity of cecal mycobiota in mice. There were significant differences in the composition of cecal fungal communities between wild and C57BL/6J mice, with more species diversity and richness of fungi in wild mice than C57BL/6J mice. We cultured 428 fungal strains from the cecal mycobiota, sequenced the whole genome of 48 selected strains, and identified 500,849 genes. Functional annotation analysis revealed multiple pathways related to energy metabolism, carbohydrate metabolism, fatty acid metabolism, and enzymes involved in the degradation of polysaccharides, lipids, and proteins, and secondary metabolite biosynthesis. The functions and abundance of and , which included the majority of the crucial metabolic pathways, were significantly higher in wild mice than in C57BL/6J mice. The results suggest that variations in the fungal community composition may relate to the adaptability of mice to their environmental habitats.

Invasive candidiasis has emerged as a significant healthcare challenge, with a rising incidence rate attributed to the widespread use of organ transplantation, chemotherapy, immunosuppressants, and broad-spectrum antibiotics. The increasing prevalence of drug-resistant strains, particularly among , has necessitated the exploration of novel therapeutic strategies. Our study investigated the synergistic effects of minocycline (MIN) combined with fluconazole (FLC) against FLC-resistant , both and . The synergistic activity of MIN and FLC was evaluated using checkerboard titration and time-kill assays. The larvae and mouse model were employed to assess efficacy, with histopathological examination and fungal burden quantification. Whole-genome and RNA sequencing elucidated the synergistic mechanisms observed. The FLC/MIN combination significantly lowered the minimum inhibitory concentration (MIC) and improved fungicidal activity, as evidenced by enhanced survival rates and reduced fungal burden in larvae and mouse models. Histopathological analysis confirmed less tissue damage and fungal load with combination therapy. RNA sequencing analysis suggested that the impact of MIN on amino acid metabolism contributes to the synergistic effects. This approach holds promise for treating FLC-resistant by increasing antifungal efficacy and reducing drug resistance risks, warranting further clinical exploration.