Antibiotic resistance is a pressing global crisis fueled by overuse in medicine, farming, and agriculture, producing superbugs like MRSA (Methicillin-resistant Staphylococcus aureus) and extended-spectrum-lactamase (ESBL) E. coli, causing prolonged and untreatable infections. Essential oils (EO) provide a natural alternative, offering broad-spectrum antibacterial potency with reduced risk of rapid resistance. Present study assessed the antimicrobial efficacy of 71 essential oils against 144 clinical bacterial isolates by determining their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) as compared with 14 standard antibiotics. Twelve EOs such as thyme, holy basil, lemon grass, cinnamon, oregano, rosewood, cassia, rose, clove, palmrosa, carom seeds and betel leaf showed remarkable potency. Out of 12 Eos, thyme, cinnamon, cassia, oregano and carom seeds were found to be most effective and categorized as strong while holy basil, lemongrass, rosewood, palmrosa, rose, clove and betel leaf were considered as moderate effective EOs. Notably, oregano achieved the lowest MIC (0.039 μL) against E. coli and the lowest MBC (0.156 μL) against Klebsiella pneumoniae. EOs demonstrates powerful antibacterial effects against multidrug-resistant pathogens, frequently surpassing conventional antibiotics in efficacy. Judicious use of these natural oils as feed additives can effectively contribute to the livestock health.
Background Human parvovirus B19 (HB19V) infection poses significant clinical challenges in resource-limited settings, particularly regarding blood safety surveillance and diagnosis of associated syndromes. Limited access to validated diagnostic assays restricts clinical management and transfusion safety protocols. Objective To develop and validate an in-house duplex real-time PCR assay for HB19V detection and evaluate its clinical utility in a resource-limited setting. Methods A duplex real-time PCR assay targeting the conserved NS1 gene of HB19V with RNase P as endogenous control was developed. Analytical validation assessed amplification efficiency, limit of detection (LOD), precision (intra- and inter-assay coefficients of variation), and cross-reactivity. Clinical validation compared qualitative agreement with a CE-certified commercial kit using Cohen's kappa statistic. Positive samples underwent Sanger sequencing for genotype characterization. Clinical specimens from suspected HB19V infections were tested and results correlated with patient presentations. Results The assay demonstrated 98.82% amplification efficiency with LOD of 6 copies per reaction. Precision metrics showed excellent reproducibility (CoV <1% intra-assay, <5% inter-assay) with no cross-reactivity. Qualitative agreement with the commercial reference was 100% (Cohen's kappa >0.95). Clinical application identified three significant cases: post-transfusion HB19V in neonatal bilirubin encephalopathy, HB19V consideration in severe anaemia with ventriculitis, and active infection in paediatric Ewing sarcoma with chemotherapy-induced cytopenias requiring IVIG therapy. Genotyping revealed genotypes 1a and 3, confirming regional viral diversity. Conclusion This validated in-house duplex real-time PCR assay provides a cost-effective, locally sustainable diagnostic tool for HB19V detection. It addresses critical gaps in blood safety surveillance and expands diagnostic capacity for HB19V-associated syndromes in resource-limited settings, while highlighting the necessity of contextual clinical interpretation for optimal patient management.
Lumpy skin disease (LSD) has emerged as a global threat to cattle health and production. Although India has been encountering regular episodes of LSD epidemics on the mainland since 2019, the Andaman and Nicobar Islands remained free of LSD until 2021. In this study, we investigated the first LSD outbreak on Great Nicobar Island in 2022 and examined the genetic characteristics of the LSDV strain associated with this outbreak. The morbidity and mortality rates in cattle were 38.29% and 1.89%, respectively. We screened 123 samples (whole blood, nasal swab, or skin tissue) from 111 cattle by LSDV real-time PCR and sera from 86 cattle by LSDV antibody ELISA, and the results confirmed LSDV infection. Subsequent nucleotide sequencing and phylogenetic analysis of five selected marker genes (GPCR, RPO30, P32, EEV, and B22R) revealed that the LSDV strain from Great Nicobar Island resembled cluster 2.5 LSDV recombinant strains from East and Southeast Asia but was distinct from wild-type LSDV strains (1.2.1, 1.2.2) circulating in mainland India, indicating an exotic source of introduction. Phylogenetic analysis using a concatenated sequence (GPCR-RPO30-P32-EEV-B22R) showed that both the wild-type and recombinant LSDV strains formed well-supported clusters, indicating that this type of analysis may be used as an alternative to whole-genome sequence analysis. We also found that determination of the nucleotide sequence of the C-terminal 717 bp of the B22R gene may be sufficient for reliable assignment of circulating LSDV isolates to a particular cluster. This is the first report of the detection of a recombinant LSDV strain in India, demonstrating the spread of cluster 2.5 recombinant LSDV further into South Asia. Our findings highlight the value of LSDV surveillance and genetic analysis for LSDV epidemiology, which may be helpful for developing effective control strategies.
As breed is known to influence metabolic traits, we compared the energy indices in the heifers of milch (Holstein Frisian, Jersey and their crosses) and draught (Amrut mahal, Hallikar, and Khillari) breeds (n = 6/breed) maintained under a subtropical production system. Intravenous insulin tolerance test and glucose tolerance test were done to the heifers and serial blood samples were collected to determine the concentration of glucose (glc), insulin, and surrogate indices of insulin sensitivity. Further, the relative expression of glucose transporters (GLUT) in the skeletal muscle, udder, and peripheral blood mononuclear cells were determined by qRT-PCR. The results revealed that the indigenous draught breeds had significantly higher glc and insulin concentrations, glycated hemoglobin, and plasma fructosamine. In contrast, exotic and crossbred dairy breeds had higher serum non-esterified fatty acid and β-hydroxybutyrate concentrations (P < 0.05). Further, the indigenous draught breeds showed a significantly higher slope and intercept of the glc curve, insulin stimulated higher blood glc reduction and elimination rate of glc than the pure and crossbred dairy breeds (P < 0.05). Insulin-stimulated glc uptake was higher in indigenous breeds, as evident from higher (P < 0.05) slope and intercept of the glc curve, glc clearance rate, and lower (P < 0.05) T1/2 of glc during intravenous glc tolerance tests. The expression of GLUT 4 was upregulated (P < 0.05) in the skeletal muscles of indigenous breeds, while GLUT 1 expression was upregulated (P < 0.05) in the udder of dairy breeds. The increased peripheral insulin sensitivity was partly due to upregulation of GLUT4 in the skeletal muscle of the indigenous draught breeds.
In January 2025, the highly pathogenic avian influenza A(H5N1) virus clade 2.3.2.1a infection was detected in domestic cats and whole-genome sequencing of two cat H5N1 isolates was performed using the Oxford Nanopore MinION sequencing platform. Phylogenetic analysis revealed the circulation of triple reassortant viruses in cats. Although cat viruses lacked classic mammalian adaptation markers they carried mutations associated with enhanced polymerase activity in mammalian cells and increased affinity for α2-6 sialic acid receptor suggesting their potential role in facilitating infection in cats. The identification of reassortant HPAI H5N1 clade 2.3.2.1a viruses in domestic cats in India highlights the urgent need for enhanced surveillance in domestic poultry, wild birds, and mammals, including humans, to track genomic diversity and molecular evolution of circulating strains. ### Competing Interest Statement The authors have declared no competing interest.
