The study was conducted with the aim to investigate the VRSA isolates in terms of their susceptibility to routinely used biocides influenced by the co-occurrence of biocide resistant gene (BRGs) and efflux pumps genes. Frequently touched surfaces within intensive care unit (ICU) of cardiac hospital were classified into three primary sites i.e., structure, machines and miscellaneous. Over a period of six months (January 2021 to July 2021) twenty three swabs samples were collected from these sites. Subsequently, these samples underwent both phenotypic and molecular methods for VRSA isolation and identification. Susceptibility and efficacy testing of biocides (benzalkonium chloride (BAC), cetrimide (CET) and chlorhexidine gluconate (CHG)) were evaluated using microdilution broth and suspension method. Furthermore, specific primers were used for singleplex PCR targeting BRGs (cepA, qacA, and qacE) and efflux pump (norA, norB, norC, sepA, mepA and mdeA) associated genes. We found that 72.2 % S. aureus demonstrate the presence of vanA or vanB genes with no significant difference among three sites (p > 0.05). cepA is the most dominant BRGs followed by qacA and qacE from structure site as compared to other sites (p < 0.05). BAC showed reduced biocide susceptibility and MIC50. There was no significant difference between presence or absence of BRGs and high MIC values of VRSA isolates from all three sites. However, efflux pump genes (EFPGs) particularly norA and norA + sepA had a significant association with BRGs and reduced biocide. BAC is the most effective disinfectant against VRSA. Proper and controlled use of BAC is required to overcome the VRSA contamination. We recommend continuous monitoring of the BRGs prevalence for better prevention of microorganism dissemination and infection control in hospitals.

Antimicrobial resistance (AMR) is a growing global health challenge, particularly in specialized tertiary care settings. Despite its significance, data on the effects of seasonal variations, demographics, and healthcare settings on AMR in Pakistan are scarce. This study aimed to evaluate the prevalence and resistance patterns of AMR and identify the key contributing factors at a cardiac hospital in Faisalabad from 2012 to 2019 to inform targeted infection control strategies. This retrospective cross-sectional analysis of 3,035 patient records adhered to STROBE guidelines. AMR profiles, including the multiple antibiotic resistance (MAR) index and antibiotic resistance genes (ARGs), were examined in relation to seasonality, hospital settings, and patient demographics. This study identified Staphylococcus aureus, Klebsiella pneumoniae, and Escherichia coli as the most prevalent pathogens, with S. aureus and K. pneumoniae classified among the ESKAPE group (Enterococcus faecium, S. aureus, K. pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.). Isolation rates were higher in inpatients (IPs) than outpatients (OPs), with peak occurrences in autumn and winter among IPs and in spring among OPs. Age and sex significantly influenced pathogen isolation rates. The multiple antibiotic resistance (MAR) index was highest for S. aureus isolates between 2017 and 2019, which showed complete resistance to vancomycin and oxacillin. Key resistance genes mecA, vanA, tetM, and aph(3') were frequently co-detected in S. aureus. Polymyxin B and colistin remained the most effective antibiotics against multidrug-resistant (MDR) strains. These findings highlight the critical need for year-round AMR surveillance, with an enhanced focus during seasonal peaks, particularly for high-risk IPs in winter and autumn. Implementing localized antimicrobial stewardship programs (ASPs) and robust infection control measures is essential to reduce the AMR burden and curb its spread in inpatient settings, especially in resource-constrained healthcare systems, such as Pakistan.

The rise of biocide- and antibiotic-resistant microbes in hospital settings demands urgent and innovative strategies to curb the spread of antimicrobial resistance (AMR). This study presents an innovative disinfectant strategy that leverages antimicrobial peptides (APep) extracted from antimicrobial-producing (AMP) strains isolated from hospital environments under intense antibiotic pressure. Unlike traditional probiotic disinfectants that rely on live bacterial strains prone to AMR, this approach directly utilizes APep from non-resistant strains with a high antioxidant potential to combat AMR and oxidative stress. Candidate strains were selected based on antimicrobial susceptibility profiling, oxidative stress assays, and screening for antimicrobial activity against hospital-acquired pathogens. The active compounds were characterized using fourier-transform infrared (FTIR) spectroscopy, high-performance liquid chromatography (HPLC), liquid chromatography–tandem mass spectrometry (LC-MS/MS), and whole-genome sequencing. Biosynthetic pathways were explored using in silico analyses, including biosynthetic gene clusters (BGCs) mapping, APep prediction, and gene interaction network analyses. Bacillus paralicheniformis UAB33 was identified as a potent producer of bacitracin B1 (UB1), exhibiting strong activity against biocides and vancomycin-resistant Staphylococcus aureus (VRSA). Genomic analysis revealed 14 BGCs, including key non-ribosomal peptides (NRPs) such as bacitracin, lichenysin, and bacillibactin. A unique pathway involving bacA, bacB, and bacC genes, along with adjacent post-translationally modified peptides (RiPP) clusters, was elucidated to enhance bacitracin synthesis. UB1-infused disinfectant wipes significantly reduce microbial loads on hospital surfaces in vitro, demonstrating a promising strategy for mitigating resistant pathogens. This strategy presents a promising approach for curbing the spread of antibiotic resistance in healthcare settings and offers a scalable and innovative solution for infection control.

Hepatitis C virus (HCV) and bacterial vaginosis (BV) coinfection generate sustained inflammation with bulk production of reactive oxygen species. They have the potency to cause hepatocellular carcinoma, vaginal apoptosis, disturb pregnancy, and influence drug treatment and follow-up. This case-control study aimed to compare the redox status in HCV and BV coinfection with respect to BV mono-infection among pregnant females (PFs). Blood samples and vaginal secretions were drawn from 75 PFs divided into three groups: coinfection (n = 25), monoinfection (n = 25), and control PFs (n = 25) who are presumed healthy subjects. Blood samples were analyzed for HCV detection based on conserved 5’ untranslated region via real-time polymerase chain reaction and hematological parameters. Markers of oxidative stress (malondialdehyde and peroxidase) and antioxidants (catalase and superoxide dismutase) were checked in plasma as well as vaginal secretions of patients among all three groups. Hematological analysis reveals that hemoglobin levels, platelets, and lymphocytes decreased significantly (p < 0.050) among the coinfection followed by mono-infection group compared to the control group. Moreover, the higher isolation frequency of pathogenic bacteria (Acinetobacter spp.) and Nugent score trend was observed among the coinfection group. Antioxidant levels were significantly lower (p < 0.050) among the vaginal secretions and blood plasma of patients having coinfection with respect to the mono-infection and control groups. While oxidative stress marker was significantly highest (p < 0.050) among vaginal secretions and blood plasma of coinfection followed by mono-infection and control group. These results validate that overall redox severity was more among the coinfection compared to the mono-infection and control groups. Redox indexes should be considered in early diagnosis and treatment of HCV and BV coinfection which may also facilitate the better treatment of hepatocellular carcinoma and vaginal apoptosis.

Post-acute sequelae of COVID-19 (PASC) affect millions of people worldwide and are increasingly recognized as a disorder of failed innate immune resolution rather than a persistent viral infection. Emerging evidence shows that residual SARS-CoV-2 antigens, host-derived alarmins, reactivated latent viruses, and mucosal microbiome-derived products from oral-nasopharyngeal and gut reservoirs sustain the chronic activation of pattern-recognition receptors, inflammasomes, and complement pathways. In parallel, deficits in specialized pro-resolving mediators, impaired efferocytosis, and persistent tissue injury prevent physiological termination of inflammation. These unresolved cues drive long-lasting epigenetic and metabolic reprogramming of hematopoietic stem cells and myeloid lineages, creating maladaptive trained immunity states characterized by hyper-responsiveness or exhaustion of these cells. Thromboinflammatory processes, including aberrant NETosis and sustained interface signalingling, further reinforce self-perpetuating inflammatory circuits. Together, these pathways give rise to reproducible molecular endotypes, including thromboinflammatory, interferon-driven, and neuroinflammatory phenotypes, which explain clinical heterogeneity. Framing PASC as a disorder of impaired immune resolution within a mucosal microbial viral context provides a unifying mechanistic scaffold for biomarker identification and host-directed therapies. This review proposes that restoring active resolution programs, rebalancing metabolic-epigenetic networks, and dismantling pathogenic innate feedback loops are promising strategies for reversing the chronic immune imprint of PASC.