An experiment was conducted to isolate, identify, and characterize the soil bacteria capable of biodegrading untreated Low Density Polyethylene (LDPE) from selected locations of municipal corporation dumping stations in Thoothukudi, Southeast coast of Tamil Nadu, India. A total of 29 distinct bacteria were isolated using Tryptic Soya Agar (TSA) after enrichment of soil samples from the selected sites. Of which, two isolates produced maximum zone of clearance during screening and formed biofilm after 21 days of incubation. The 16 S rRNA sequencing confirmed that the isolates were identified as Paenibacillus timonensis (PX106822) and Cupriavidus nantongensis (PX106762) which resulted in the weight reductions of 3.15% and 1.83% respectively on untreated LDPE after 30 days of incubation. During degradation, the pH of the Mineral Salt Medium (MSM) increased from 6.73 to 8.06 and 8.11 respectively by the two isolates. In the present study, it was observed that the two isolates had increased viable cell count of 7.07 × 108 and 3.83 × 107 CFU/ml respectively with the initial attachment onto the LDPE films followed by biofilm formation, which favoured biodegradation of LDPE. Fourier Transform Infrared (FTIR) spectroscopy and Field Emission Scanning Electron Microscopy (FE-SEM) analysis also confirmed that the two isolates had adhered and attacked on the surface of LDPE film thus serving as evidence for bacterial degradation of LDPE. This study concluded that the isolates had more capability in LDPE biodegradation and will serve as a baseline data for plastic litter management.
Microorganisms play pivotal roles in maintaining host physiology and ecosystem balance, with fish-associated microbiomes offering unique insights due to the diverse habitats and feeding behaviours of their hosts. This review comprehensively explores the diversity, composition, and functional roles of gut and skin-associated microbial communities in fish across freshwater, brackish, and marine environments, with emphasis on recent advancements in metagenomic methodologies. Culture-independent techniques, particularly high-throughput and third-generation sequencing technologies, have revolutionized our ability to uncover microbial diversity, gene functions, and interspecies interactions. The fish gut microbiome, heavily influenced by factors such as diet, habitat, and host species, contributes significantly to nutrient metabolism, immune modulation, and physiological adaptation. Similarly, the skin microbiota provides a critical first line of defence, offering protection through competitive exclusion and antimicrobial activity. Functional metagenomics reveals microbial contributions to host metabolism, energy homeostasis, xenobiotic degradation, and environmental adaptation via the gut-brain axis and metabolic pathways. Emerging evidence highlights the bidirectional relationships between microbiota and host phenotypic plasticity. This review underscores the importance of integrative metagenomic approaches to decode complex microbial functions and their ecological relevance in aquaculture, with implications for sustainable fish health management, disease prevention, and improved productivity.
Diatoms, a unique class of microalgae, play a pivotal role in global ecosystems due to their photosynthetic capabilities and adaptability to diverse aquatic environments. Composed of opaline silica, they serve as sensitive indicators for water quality monitoring. This review covers various diatom research techniques, including sampling and isolation. It highlights methods for different substrata, such as epiphyton, epipelon, and epipsammon, as well as traditional isolation approaches like agar plate methods and serial dilution. Automation techniques, such as flow cytometry (FACS), are also discussed. The review emphasizes the potential of diatoms for pollutant removal, particularly heavy metals, and presents various diatom-based indices for water quality assessment. Future research is encouraged to refine isolation techniques, enhance the effectiveness of indices, and explore biochemical pathways that could improve diatoms’ bioremediation capabilities, thereby fully leveraging their potential for environmental monitoring and remediation.
Food-borne bacterial pathogens remain a major public health concern, causing extensive illness and economic losses worldwide. Conventional detection methods are often slow and insufficient for identifying viable but non-culturable pathogens. Recent microbiological, biotechnological and bioinformatic advances have markedly improved food safety monitoring. Rapid molecular assays (PCR, qPCR, microarrays), next-generation sequencing, metagenomics, and emerging CRISPR-based diagnostics enable faster and more accurate pathogen detection and outbreak tracing. Bioinformatic tools—including genomic databases, phylogenetics, and machine-learning models—support predictive risk assessment and real-time surveillance. Preventive innovations such as bacteriophages, probiotics, antimicrobial peptides, nanotechnology-based interventions, and engineered microbes provide sustainable alternatives to chemical preservatives. Key challenges include variability across food matrices, biosafety considerations, and limited integration of multi-omics approaches into routine workflows. Overall, these emerging strategies offer improved precision and responsiveness for detecting and preventing food-borne bacterial pathogens.
The emergence of CRISPR/Cas9 technology has transformed the landscape of gene editing, allowing for precise alterations in DNA that hold great promise for research and potential therapies. However, a significant concern is the occurrence of off-target effects, which can lead to unintended genetic modifications with potentially harmful consequences. This paper explores the nature of off-target effects in CRISPR/Cas9, discussing how they arise and their implications for the reliability of gene editing. We identify the challenges faced in detecting and predicting these off-target interactions, including limitations in current detection techniques and the complexities of cellular biology. We present strategies aimed at minimizing off-target effects, such as careful design of guide RNAs, the use of computational tools for prediction, and improved delivery methods. Through a review of case studies, we highlight successful cases where off-target activity has been significantly reduced, offering insights into best practices for enhancing the accuracy of CRISPR/Cas9 applications. Moreover, we provide a comparative overview of Cas9, Cas12, and Cas13 systems, emphasizing their distinct target specificities, mechanisms of action, and off-target profiles. This comparison offers a broader understanding of how alternative CRISPR effectors may be leveraged to improve genome and transcriptome editing precision. This study underscores the importance of continued research to address the challenges of off-target effects, ultimately supporting the development of safer and more effective gene editing methods for clinical use.
The rise of multidrug-resistant (MDR) bacteria, especially the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.), is one of the most significant issues in modern medicine. Berberine, a quaternary ammonium isoquinoline alkaloid derived from a number of plant species, has been shown to be an effective therapeutic agent against resistant pathogens. This review provides a detailed overview of the chemical structure, pharmacology, and mechanism of action for berberine against ESKAPE and other MDR bacteria. The literature suggests that berberine displays antimicrobial activity through several mechanisms, including damaging the membrane, inhibiting DNA gyrase, inhibiting protein synthesis, and inhibiting efflux pumps. Berberine shows considerable synergy when combined with standard antibiotics, which could reverse antibiotic resistance. Notably, berberine demonstrates synergistic effects with β-lactam antibiotics, reducing fractional inhibitory concentration (FIC) indices to 0.25–0.5, thereby enhancing antibacterial efficacy against multidrug-resistant strains. Although berberine exhibits remarkable in vitro antimicrobial activity, its very poor systemic bioavailability (< 1%) results in a more than 1000-fold PK–PD gap between achievable plasma levels and effective MIC values. The clinical and preclinical studies show a good safety profile for use with minimal toxicity at therapeutic concentrations. Therefore, the current clinical use of berberine is limited to adjuvant, topical, or gastrointestinal applications rather than systemic monotherapy. The routes of delivery, pharmacokinetic characteristics, and standardization of treatment remain obstacles for wider application. This review collates the current evidence that supports berberine as an alternative or adjunct therapy to combat the worldwide issue of antibiotic resistance and also indicates areas where further research is required to bring therapies to the clinical level.
Leptospirosis is a globally significant zoonotic disease caused by the pathogenic Leptospira spp. This study aimed to develop a recombinant Loa22 based lateral flow assay for detecting leptospirosis in dogs and cattle. The loa22 gene from Leptospira interrogans was cloned, expressed in E. coli, and purified using Ni–NTA affinity chromatography. The immunoreactivity of the recombinant protein was confirmed by western blotting, and hyperimmune sera against the protein was raised in mice. Gold nanoparticles (AuNP) were conjugated with rLoa22 under optimized conditions and characterized by UV–Vis spectrophotometry, agarose gel electrophoresis, and transmission electron microscopy. The lateral flow assay (LFA) strips were assembled with AuNP–rLoa22 on the conjugate pad and rLoa22 and hyperimmune sera on the test and control lines, respectively. Diagnostic performance of the assay was evaluated using sera from 100 dogs and 102 cattle suspected for leptospirosis and compared to the reference standard microscopic agglutination test (MAT). Six dog and cattle samples each were tested positive by MAT. The developed LFA further tested 06 dog and 07 cattle samples as positive in addition to the MAT positive samples, representing false positives. In dogs, the LFA showed 100% sensitivity and 93.62% specificity, while in cattle, it showed 100% sensitivity (Se) and 92.71% specificity (Sp) with respect to MAT. The n/N for Se and Sp were, Sedogs= 6/6; Spdogs= 88/94; Secattle= 6/6; Spcattle= 89/96. The high observed sensitivity could be attributed to the smaller number of MAT+/LFA + samples that resulted in a wide 95% confidence interval (CI). The test was further validated on another 300 canine and 98 cattle serum samples suspected for leptospirosis (prevalence screens, not a part of the accuracy cohorts). The results underscore the potential of the rLoa22–based LFA as a reliable, rapid diagnostic tool for leptospirosis screening in veterinary settings. Further validation of the study is warranted using samples pre-screened for leptospirosis through other commercially available techniques, including qPCR and ELISA.
Emerging contaminants are molecules, either novel or previously recognized, that persist in the environment and may pose risks to ecosystems and human health. Their increasing occurrence, particularly in pharmaceuticals, personal care products, and industrial processes, has intensified research on their detection, monitoring, and ecological impact. Advances in analytical technologies now enable the identification of these compounds at trace concentrations, yet their long-term effects remain uncertain. This review compiles recent findings on microbial ecotoxicology, focusing on representative contaminants of high concern such as penicillin, parabens, caffeine, and microplastics. Microorganisms (bacteria or fungi) are highlighted both as sensitive bioindicators of environmental pollution and as active agents in biodegradation processes. Their ability to metabolize, transform, or neutralize contaminants underscores their potential as sustainable tools for remediation. By integrating evidence from multiple studies, we emphasize microbial-based strategies as promising tools for environmental monitoring and mitigation.
This study investigates the interactions between Pestalotiopsis diospyri and three phytopathogenic fungi—Fusarium guttiforme, Colletotrichum horii, and C. gloeosporioides—isolated from papaya and pineapple. A comparative analysis of metabolite production using 1H NMR was conducted under axenic and co-culture conditions with rice and potato dextrose broth (PDB) media. Principal Component Analysis (PCA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) of the 1H NMR data revealed metabolic trends between axenic and co-culture conditions, while compound-specific PCA demonstrated distinct species-based metabolite clustering, supporting the observed metabolic changes during interspecies interactions. The results revealed various classes of metabolites in the axenic cultures; however, in the tri-cultures, α-pyrones synthesized exclusively by P. diospyri were differentially expressed, with compounds 11 and 12 showing up to 2-fold increases in yield. The diverse survival strategies exhibited by these fungi, including antagonistic and parasitic behaviors, notably influenced their interactions, leading to the up-regulation and down-regulation of specific metabolites, particularly α-pyrones. Comparative ¹H NMR profiling indicated that the metabolic response to co-culture was dominated by known α-pyrones, with no detectable emergence of uncharacterized metabolites that might signal the production of potent toxins. Furthermore, our findings indicate that the rice medium enhances substrate-fungi interactions, thereby increasing both chemo-diversity and metabolite yield. Preliminary antifungal assays revealed that the α-pyrones exhibited weak to moderate, strain-specific activity against the phytopathogenic fungi, suggesting their ecological role in interspecies competition rather than as potent antimicrobial agents.
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