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Mystus cavasius, known as Gangetic mystus, is a freshwater indigenous catfish. The present study represents the first-ever complete mitochondrial genome sequencing of M. cavasius. The whole mitochondrial genome size is 16,554 bp (GenBank accession number OR018997). The mitogenome organization consists of total 37 genes, with 13 protein-coding genes, two rRNA, 22 tRNAs, and a D-loop regulatory region. Among these, 28 genes are coded on the heavy strand, while eight tRNA genes and ND6 are encoded separately. Phylogenetic analysis reveals that M. cavasius clusters with other Mystus species and bagrid catfishes. This work contributes valuable insights into structural characterization and phylogenetic relationships.

Aquaculture is one of the major economic activities in India, providing livelihoods and nutritional security to millions of people. In recent times, fish diseases have come to the limelight resulting in significant economic losses. We aimed to identify pathogenicity and virulence profiling of thirty-six pathogenic bacterial strains isolated from diseased Labeo rohita in the district of Hooghly, West Bengal, India. The bacterial strains were characterized through a comprehensive approach involving the examination of morphological features, biochemical properties, amplification, and sequencing of the 16S rRNA, species-specific genes, and virulence genes. Considering the prevalence frequency, virulence potential, and statistical significance Aeromonas hydrophila and Pseudomonas aeruginosa were selected for a survival assay followed by the examination of histopathological features to elucidate their effects. The identified bacterial isolates were arranged based on their predominance frequency, i.e., Aeromonas hydrophila (25%), Aeromonas veronii (22%), Pseudomonas aeruginosa (22%), Enterococcus faecalis (14%), Klebsiella pneumoniae (6%), Staphylococcus aureus (6%) and Escherichia coli (5%). Sixteen virulence-associated genes related to pathogenicity were amplified across the thirty-six isolates; aer, alt, lip and hlyA for A. hydrophila; exoS, lasB, toxA, oprL and phzM for P. aeruginosa; entB, fimH and uge in K. pneumoniae; aer in A. veronii; hlyA in E. coli; hlb in S. aureus and gelE for E. faecalis. The log-probit analysis revealed that A. hydrophila was notably more pathogenic than P. aeruginosa, as indicated by its lower lethal dose of 1.5×104 CFU/mL. Additionally, histological examination revealed notable pathological changes, including tissue degeneration, inflammatory cell infiltration and vacuolation observed in the liver, kidney, gill and intestine of the challenged fish. We highlighted several potent aquatic microbial pathogens in order to manage and prevent such aquacultural maladies.

Abstract The Sundarbans mangrove, located at the mouth of the Ganges and Brahmaputra Rivers, is the world’s largest tidal mangrove forest. These mangroves are also one of the most striking sources of microbial diversity, essential in productivity, conservation, nutrient cycling, and rehabilitation. Hence, the main objective of this study was to use metagenome analysis and provide detailed insight into microbial communities and their functional roles in the Sundarbans mangrove ecosystem. A comparative analysis was also done with a non-mangrove region of the Sundarbans ecosystem to assess the capability of the environmental parameters to explain the variation in microbial community composition. The study found several dominant bacteria, viz., Alphaproteobacteria, Actinomycetota, Bacilli, Clostridia, Desulfobacterota, Gammaproteobacteria, and Nitrospira, from the mangrove region. The mangrove sampling site reports several salt-tolerant bacteria like Alkalibacillus haloalkaliphilus, Halomonas anticariensis, and Salinivibrio socompensis. We found some probiotic species, viz., Bacillus clausii, Lactobacillus curvatus, Vibrio mediterranei and Vibrio fluvialis, from the Sundarbans mangrove. Nitrifying bacteria in Sundarbans soils were Nitrococcus mobilis, Nitrosococcus oceani, Nitrosomonas halophila, Nitrospirade fluvii, and others. Methanogenic archaea, viz., Methanoculleus marisnigri, Methanobrevibacter gottschalkii, and Methanolacinia petrolearia, were highly abundant in the mangroves as compared to the non-mangrove soils. The identified methanotrophic bacterial species, viz., Methylobacter tundripaludum, Methylococcus capsulatus, Methylophaga thiooxydans, and Methylosarcina lacus are expected to play a significant role in the degradation of methane in mangrove soil. Among the bioremediation bacterial species identified, Pseudomonas alcaligenes, Pseudomonas mendocina, Paracoccus denitrificans, and Shewanella putrefaciens play a significant role in the remediation of environmental pollution. Overall, our study shows for the first time that the Sundarbans, the largest mangrove ecosystem in the world, has a wide range of methanogenic archaea, methanotrophs, pathogenic, salt-tolerant, probiotic, nitrifying, and bioremediation bacteria.

IntroductionThe Brahmaputra, a major transboundary river of the Himalayas flowing predominantly through Northeast India, particularly Assam, is increasingly endangered by contamination due to rapid urbanization and anthropogenic pressures. These environmental changes pose significant risks at the microbial level, affecting nutrient cycling and productivity, and thereby impacting river ecosystem health. The next-generation sequencing technology using a metagenomics approach has revolutionized our understanding of the microbiome and its critical role in various aquatic environments.MethodsThe present study aimed to investigate the structure of the bacterial community and its functional potentials within the sediments of the Brahmaputra River, India, using high-throughput shotgun metagenomics. Additionally, this study sought to explore the presence of antimicrobial resistance genes in the river’s sediment.Results and discussionShotgun metagenomics revealed a diverse bacterial community comprising 31 phyla, 52 classes, 291 families, 1,016 genera, and 3,630 species. Dominant phyla included Pseudomonadota (62.47–83.48%), Actinobacteria (11.10–24.89%), Bacteroidetes (0.97–3.82%), Firmicutes (0.54–3.94%), Cyanobacteria (0.14–1.70%), and Planctomycetes (0.30–0.78%). Functional profiling highlighted significant involvement in energy metabolism, amino acid and central carbon metabolism, stress response, and degradation pathways, emphasizing the microbial community’s role in ecosystem functioning and resilience. Notably, 50 types of antibiotic resistance genes (ARGs) were detected, with resistance profiles spanning multidrug, aminoglycoside, β-lactam, fluoroquinolone, rifampicin, sulfonamide, and tetracycline classes. Network analysis underscored the intricate relationships among ARG subtypes, suggesting potential mechanisms of resistance propagation. Furthermore, plasmid-related genes and 185 virulence factor genes (VFGs) were identified, indicating additional layers of microbial adaptation and potential pathogenicity within the river sediments. This comprehensive microbial and functional profiling of the Brahmaputra’s sediment metagenome provides crucial insights into microbial diversity, resistance potential, and ecological functions, offering a foundation for informed management and mitigation strategies to preserve river health and mitigate pollution impacts.

Abstract Background Shallots are infected by various viruses like Onion yellow dwarf virus (OYDV), Leek yellow stripe virus (LYSV), Shallot latent virus (SLV) and Shallot virus X (ShVX). In India, they have been found to be persistently infected by ShVX. ShVX also infects onion and garlic in combination with other carlaviruses and potyviruses. ShVX is a member of genus Allexivirus of family Alphaflexiviridae. ShVX has a monopartite genome, which is represented by positive sense single-stranded RNA. Globally, only six complete and 3 nearly complete genome sequences of ShV X are reported to date. This number is insufficient to measure a taxon’s true molecular diversity. Moreover, the complete genome sequence of ShVX from Asia has not been reported as yet. Therefore, this study was undertaken to generate a complete genome sequence of ShVX from India. Results Shallot virus X (ShVX) is one of the significant threats to Allium crop production. In this study, we report the first complete genome sequence of the ShVX from India through Next-generation sequencing (NGS). The complete genome of the ShVX (Accession No. OK104171), from this study comprised 8911 nucleotides. In-silico analysis of the sequence revealed variability between this isolate and isolates from other countries. The dissimilarities are spread all over the genome specifically some non-coding intergenic regions. Statistical analysis of individual genes for site-specific selection indicates a positive selection in NABP region. The presence of a recombination event was detected in coat protein region. The sequence similarity percentage and phylogenetic analysis indicate ShVX Indian isolate is a distinctly different isolate. Recombination and site-specific selection may have a function in the evolution of this isolate. This is the first detailed study of the ShVX complete genome sequence from Southeast Asia. Conclusion This study presents the first report of the entire genome sequence of an Indian isolate of ShVX along with an in-depth exploration of its evolutionary traits. The findings highlight the Indian variant as a naturally occurring recombinant, emphasizing the substantial role of recombination in the evolution of this viral species. This insight into the molecular diversity of strains within a specific geographical region holds immense significance for comprehending and forecasting potential epidemics. Consequently, the insights garnered from this research hold practical value for shaping ShVX management strategies and providing a foundation for forthcoming studies delving into its evolutionary trajectory.

Despite major advances in biosensing, quick, dependable, and effective on-site detection of bacterial infections remains a serious issue, owing to a lack of acceptable or appropriate diagnostic platforms. To address this gap, we presented a new colorimetric gold NanoZyme aptasensor for rapid sensing of Aeromonas veronii, an infectious bacterial disease in fish. The A. veronii-specific aptamer (AVS01) was developed through Cell-SELEX. The sensing mechanism involves inhibition of AuNPs induced peroxidase-mimic catalytic activity through surface adsorption by AVS01 which in the presence of the A. veronii desorb from the AuNPs allowing recovery of the catalytic activity leading to colorimetric response, whereas the sensor is insesnsitive to other nontarget bacterial cells. This method is very specific and sensitive, allowing for the quick and visible sensing of A. veronii with a detection limit of 1281 CFU mL−1 within 15 min. The method has great potential for rapid diagnosis of bacterial infection in fish caused by A. veronii.

Abstract Background Chickpea (Cicer arietinum L.) production is affected by many biotic factors, among them Fusarium wilt caused by Fusarium oxysporum f. sp. ciceri and Botrytis gray mold caused by Botrytis cinerea led to severe losses. As fungicide application is not advisable, biological management is the best alternative for plant protection. The rhizosphere-dwelling antagonistic bacteria are one of the important successful alternative strategy to manage these diseases of chickpea. Rhizosphere dwelling bacteria serve as biocontrol agents by different mechanisms like producing antibiotics, different enzymes, siderophores against pathogens and thereby reducing the growth of pathogens. Results The present study aimed to isolate rhizospheric bacteria from the soils of different chickpea fields to evaluate biocontrol efficacy of the isolated bacteria to manage Fusarium wilt and Botrytis gray mold in chickpea. A total of 67 bacteria were isolated from chickpea rhizosphere from Bundelkhand region of India. Study revealed the isolated bacteria could reduce the Fusarium oxysporum f. sp. ciceris and Botrytis cinerea infection in chickpea between 17.29 and 75.29%. After screening of all the bacteria for their biocontrol efficacy, 13 most promising bacterial isolates were considered for further study out of which, three bacterial isolates (15d, 9c and 14a) have shown the maximum in vitro antagonistic effects against Fusarium oxysporum f. sp. ciceri and Botrytis cinerea comparable to in vivo effects. However, Isolate (15d) showed highest 87.5% and 82.69% reduction in disease against Fusarium wilt and Botrytis gray mold respectively, under pot condition. Three most potential isolates were characterized at molecular level using 16S rRNA gene and found to be Priestia megaterium (9c and 14a) and Serratia marcescens (15d). Conclusion This study identified two native biocontrol agents Priestia megaterium and Serratia marcescens from the rhizospheric soils of Bundelkhand region of India for control of Fusarium wilt, Botrytis gray mold. In future, efforts should be made to further validate the biocontrol agents in conjugation with nanomaterials for enhancing the synergistic effects in managing the fungal diseases in chickpea. This study will definitely enhance our understanding of these bioagents, and to increase their performance by developing effective formulations, application methods, and integrated strategies.

Fusarium stalk rot (FSR), caused by the Fusarium species complex, is an economic threat to maize cultivation all over the world. We investigated the population structure and genetic diversity of Fusarium species obtained from five major maize-growing regions in India. The Tef-1α locus was used for phylogenetic analysis of geographically distinct isolates of Fusarium verticillioides, F. andiyazi, F. proliferatum, F. nygamai, and F. acutatum causing FSR. A phylogenetic tree showed monophyletic, polyphyletic, and paraphyletic groupings reflecting the complex evolutionary history and genetic diversity within the genus. Monophyletic groupings depicting strong bootstrap support were shown to have a single common ancestor and genetic coherence with limited genetic divergence among sequences. Polyphyletic groupings also presented significant genetic differentiation within the F. verticillioides sequences from diverse ecological niches. Nucleotide diversity of moderate level 0.02471 reflected genetic variations within populations that were attributed to factors such as mutation, genetic drift, or varying selection pressures. The Fst value of 0.98205 is particularly indicative of high genetic differentiation, implying that most of the genetic variance is due to differences between populations rather than within them. F. verticillioides, with 57 sequences, showed low genetic diversity with three segregating sites and a low haplotype diversity of 0.19486, suggesting the founder effect, where a reduced population expands from a limited genetic pool. The total data estimates across all populations for haplotype analysis showed 72 sequences, 44 segregating sites, and 9 haplotypes with a haplotype diversity of 0.48513. The evolutionary dynamics showed genetic differentiations among Fusarium species causing FSR. AMOVA indicated high within-population variations, depicting a substantial genetic diversity within individual populations. The results offer a comprehensive framework for discussing the implications of genetic diversity in pathogen management and the evolutionary dynamics of the Fusarium species causing FSR in maize in the Indian subcontinent.

In this study, we evaluated gamete quality parameters of mature male koi carp (Cyprinus carpio) exposed to three different concentrations (1, 10, and 100 µg/L) of di-(2-ethylhexyl) phthalate (DEHP). After 60 days of exposure, there was a significant decrease in the gonadosomatic index (GSI) of males exposed to 10 and 100 µg/L of DEHP. Histological analysis of the testes revealed impaired histoarchitecture, including inflammatory cells, intratubular vacuoles, and swollen seminiferous tubules in treatment groups. Gamete quality parameters like sperm production, motility, spermatocrit, and sperm density values were significantly decreased at the 10 and 100 µg/L concentrations. Biochemical compositions, including glucose, cholesterol, and total protein levels, were significantly changed in the treatment groups. Similarly, the ionic compositions of seminal fluid (Na, K, Ca, and Mg) also varied in the treatment groups. Furthermore, the 11-ketotestosterone levels were decreased, and the 17-β estradiol levels were increased in the DEHP-treated groups. The mRNA expression levels of reproduction-related genes, including Fshr, Lhr, Ar, Erα, and Erβ, were significantly changed in the DEHP-treated males in a dose-dependent manner. In conclusion, the findings of this study confirmed that environmentally relevant exposure to DEHP may contribute to a decline in the gamete quality of male fishes.

India is the habitat of many freshwater fish species that make a significant contribution to endemism. However, the native species population is declining at a very fast rate due to pollution, habitat loss or degradation, new species introduction, hydrologic alterations, and overfishing. The minor carp, Labeo gonius, is one of the important fish species in the Indian river systems and holds enormous potential in culture fisheries. Therefore, the precise status of population structure is a prerequisite for the management of the species. In the present study, we assessed the potential of mtDNA ATPase 6/8 and D-loop regions as markers to determine the genetic diversity and phylogeography of L. gonius from Indian rivers. We have selected five major Indian rivers for investigation which could represent geographically distant populations of the studied fish. A total of 210 ATPase 6/8 sequences (842 bp) and 203 D-loop region sequences (683 bp) of L. gonius from the five major Indian rivers were submitted to NCBI. Results revealed that ATPase 6/8 has 17 haplotypes and 70 polymorphic sites with an overall haplotype diversity of 0.560 ± 0.036 and nucleotide diversity of 0.00347 ± 0.00. Furthermore, D-loop analysis revealed 58 haplotypes, 67 polymorphic sites, 0.954 ± 0.006 overall haplotype diversity, and 0.01914 ± 0.00 nucleotide diversity. The hierarchical AMOVA analysis indicated that a high proportion of the genetic variation (76.46% and 70.22% for the D-loop and ATPase 6/8 regions, respectively) was attributed to among-population variation under gene pool-1 (considering all the stocks from Ganga, Brahmaputra, Teesta, Narmada, and Mahanadi as a single pool). Furthermore, in gene pool-2, samples were divided into two groups: group 1 (Himalayan rivers)—Ganga, Brahmaputra, and Teesta; and group 2 (peninsular rivers)—Narmada and Mahanadi. Genetic variations in the D-loop and ATPase 6/8 regions were found among populations within groups at 70.65% and 53.48%, respectively. Haplotype network analysis revealed that there were significant differences in distributions and frequencies. The observed unique haplotypes might be useful for differentiating the stocks. In brief, the present study suggested that the D-loop and ATPase 6/8 regions are potential markers and represent a significant contribution to the genetic characterization of the minor carp L. gonius from major Indian rivers. Moreover, the L. gonius population along the east coast especially in the Himalayan rivers (Ganga, Brahmaputra, and Teesta river basin) is found to be genetically more diverse. The results would provide a scientific basis for the assessment of fisheries resources and management of this important species.

Spermiation, an act of sperm release, depends on several molecular factors. Despite hormonal administration, spermiation failure is a primary concern in certain fishes. In this study, the molecular mechanisms of spermiation have been analyzed in Cyprinus carpio by comparative transcriptomics. Unigenes for C. carpio control (CCC), which were injected with PBS (Phosphate-buffered saline), and C. carpio treated (CCT), which were injected with ovatide, were 107,616 and 133,435, respectively. A total of 93 genes were identified as involved in the spermiation process, including those related to gonadal steroidogenesis, cell growth, cell adhesion, and cytoplasmic matrix formation. The cd63, CENPS, rasa1a, and genes for gonad steroidogenesis, cell growth, cell adhesion, and cytoplasmic matrix formation were analyzed. Gene expression analysis revealed tubulobulbar complexes mediated disengagement of spermatozoa and JAK2 signaling regulated cyst breakage in teleost for the first time. Analysis was done from the changes at the molecular level to the final act of spermiation. Tissue histology analysis was conducted in accordance with the molecular study, which showed structural changes. Induced breeding in fish plays a key role in seed production in aquaculture sector. However, there are several constraints the sector is still facing due to lack of extensive knowledge regarding the mechanisms of spermiation and species-specific response to hormonal dosage. This study is relevant to understand the molecular mechanisms involved in spermiation and the stages which mark as critical point of sperm release after administrating the inducing agent. This study also lays the groundwork for further exploration of species-specific responses to hormonal treatments, aiding sustainable seed production in the fisheries sector.

Plastic waste has become a significant global ecological concern due to its substantial harmful impacts. They are eventually colonized and transformed by diverse microbial communities known as the ‘‘plastisphere”. Only a limited group of microbes have been identified so far due to the prevalence of non-culturable strains that inhabit polymer substrates. We investigated the community diversity and predictive functionality of the plastisphere microbiota using a high-throughput next-generation sequencing (NGS) approach to unveil their plastic degrading potential. Samples were collected from four different plastic-enriched disposal sites adjacent to Kolkata, West Bengal, India, followed by processing, gDNA extraction, library preparation, and sequencing. The most prevalent phyla were Proteobacteria, followed by Bacteroidetes, Actinobacteria, Cyanobacteria, Firmicutes, and Verrucomicrobia. The functional profiling revealed that genes associated with metabolism, cellular processes, and signalling were most prevalent, followed by poorly characterized information storage and processing. In this study, we predicted the existence of a beneficial microbiome associated with bioremediation and plastic degradation, which suggests the potential utilization of plastics as primary carbon sources. Our findings also highlighted the existence of promising microbial enzymes associated with the biodegradation of several plastic substrates.Furthermore, our research unveiled the enriched distribution of beneficial microbiome in the studied metagenome, which offers a diverse prospect. This investigation establishes a connection between the structure of microbial communities and diverse genes actively engaged in the biodegradation of plastic waste within plastic disposal sites. These beneficial degraders can be investigated further for broad-spectrum applications in plastic bioremediation. Subsequent explorations of their plastic degrading enzymes will provide profound contributions to plastic pollution mitigation.

Hexavalent chromium or Cr(VI) is highly toxic for humans as it causes high oxidative reactions inside cells, leading to diseases like chronic ulcers and damage to the kidneys, mucous membranes, throat, skin, and respiratory tract. Due to rapid urbanization, Cr(VI) comes into our food chain through unmonitored and uncontrolled application in agriculture fields, refineries, mills, the tanning industry, automobiles, road works, etc. Presently, the standard Cr(VI) detection is done using conventional processes, which, though accurate, has severe drawbacks in on-the-spot rapid detection in the field. Here, we have represented a handheld potentiostat towards trace-level Cr(VI) detection in aquatic environments. A Cr(VI) specific DNA aptamer immobilized screen printed electrode (SPE) has been used as the main biosensor. The device operates on an electronic peak current dumping event through mass deposition in the presence of an aptamer coupled with Cr(VI) onto the working electrode. The working range of the developed prototype is in the range of 0–1000 ppb Cr(VI), where the maximum linearity has been observed in the range of 0–500 ppb with a limit of Detection (LOD) as low as 10 ppb. The device has exhibited an excellent correlation with commercially available electrochemical workstations with a coefficient of 0.972. Moreover, the applicability of the developed device has been validated for 7 different types of water samples. To our knowledge, this is the first-ever reported simplistic resource-limited on-spot aptasensing device for Cr(VI) detection in an aquatic environment.

The river Ganges in India has faced considerable issues due to water quality degradation caused by various anthropogenic activities. This study employs metagenomic analysis to comprehensively characterize bacterial communities, explore functional genomics, and investigate the prevalence of virulence factors and antibiotic resistant genes (ARGs) within the sediment environment of the river Ganges. Taxonomic profiling revealed that Proteobacteria were the most dominating phyla found in all samples, whereas the abundance of Pseudomonas at the genus level was the highest in all the samples. Functional annotation and pathway analysis uncover the genomic potential of sediment associated bacteria, shedding light on metabolic pathways, biogeochemical processes, human diseases and adaptive mechanisms within the riverine ecosystem. Moreover, identifying the highest number of genes related to virulence factors was observed in K1 samples (3), and the highest number of genes related to ARGs found in K3 (25) samples emphasizes the need to understand potential pathogenicity in these environments. Characterization of ARGs provides crucial insights into the prevalence of resistance determinants, their genetic contexts, and potential sources of antibiotic resistance in this vital aquatic ecosystem. Overall metagenomics analysis in different sampling sites of river Ganga observed nearly the same OTUs at microbial communities at the taxonomic level but not at a functional level. This research can be a critical foundation for assessing the ecological implications of microbial communities, functional genomics, and the resistome in the river Ganges sediments. The findings underscore the importance of metagenomic approaches in elucidating the intricate microbial ecology and the prevalence of genetic elements relevant to environmental health and antimicrobial resistance in aquatic ecosystems. Further investigation would be required to understand the underlying cause behind the restoration of microbial functional profiles, including ARGs and VFs, to unravel the rejuvenation aspects of this unique ecosystem.

To delineate the response mechanism of Nile tilapia (Oreochromis niloticus) to the riverine pollution of river Ganga, a comparative transcriptomic analysis was performed utilizing fresh liver through RNA-Seq technology. A total of 51.39 million and 32.8 million reads were obtained after excluding low quality sequences from non-polluted (Barrackpore) and polluted (Kanpur) sites of Nile tilapia. About 81.4 % and 95.3% reads were perfectly mapped with the reference sequence of O. niloticus. Transcriptional analysis generated 363 differential expressed genes (DEGs) including 131 up-regulated and 232 down-regulated genes. Gene Ontology (GO) enrichment analysis revealed that significant DEGs were associated with ribosome biogenesis, alpha-amino acid metabolic process, translational initiation etc. as biological process (BP); unfolded protein binding, vitamin binding, carboxylic acid binding and etc. as molecular function (MF); ribosome, and ribosomal subunit as cellular component (CC). The KEGG analysis indicated that these DEGs were highly involved in ribosome, Lysine degradation and RNA transport pathways. Additionally, ten hub genes participated in Translation, Ribonucleoprotein complex biogenesis as BP, Ribosome, Eukaryotic translation initiation factor 3 complex as CC and Structural constituent of ribosome, RNA binding as MF were affected in riverine pollution. Overall, this transcriptome investigation provided an extensive overview of pollution triggered transcriptional mechanisms in Tilapia liver and would be highly significant for further exploration of the molecular processes in response to pollution.

The current study aimed at assessing the immunostimulatory properties of silver nanoparticles (AgNPs) on Labeo rohita, and understanding how it affects the growth, cellular ultrastructure, the expression level of immune genes, and infection risk from Aeromonas hydrophila. Fish (avg wt: 30.1±3.26 g) were fed diets with four separate AgNP inclusion levels (0 µgKg−1 [basic diet, T1], 10 µgKg−1 [T2], 15 µgKg−1 [T3], and 20 µgKg−1 [T4]) for 56 days. After the feeding trial, growth, histological, immunological parameters, and protective immune response against A. hydrophila were assessed. The fish in the treatment groups including T1(control), the T3 growth indices, such as specific growth rate (7.56±0.26) and percent weight gain (231.05±3.21), was statistically higher (P < 0.05). In the immunological and oxidative parameters, levels of SOD and catalase decreased in correlation with a rise in the inclusion doses of AgNP in the liver, and a reduction in catalase values was recorded in the gill. With the addition of AgNP, the NBT value was decreased in the gill, and T3 had a considerably larger (P

Wetlands are recognized as one of the most productive aquatic ecosystems. This high productivity supports its enormous biodiversity, aiding a scope for the small scale fisheries within it. Basically, these wetlands act as platforms for culture-based fisheries (CBF) i.e practice of stocking fish seeds and harvesting the grow out fishes at later. Often in tropical wetlands, the high nutrient load and high temperature enable the luxurious growth of macrophytes. Moreover, this macrophyte infestation is considered as a bottleneck for wetland managers. Facing a similar situation our studied Beledanga wetland is a macrophyte-infested wetland and the invasive Eichhornia crassipes is the most dominant macrophyte species. Owing to the macrophyte problem, Grass carp have been stocked in it as a biocontrol agent, as grass carp is a known biocontrol agent for the macrophyte infestation situation and simultaneously it will raise the fish production. To ensure maximum sustainable production from the wetland by harnessing with the available resources, it is necessary to acquire knowledge about the system’s ecological status and its ecological networks. Thus, the Ecopath based mass-balanced model is one of the best suitable tools to know about the ecological organization of this Beledanga wetland. Considering the available data (observational and literature data), this study has constructed a mass-balanced Ecopath model with 14 functional groups including all fish groups, birds, plankton, benthos and detritus. It has been found that the zooplankton and benthos are the most utilized group and the birds are the keystone group in this wetland system. This wetland possesses a lower transfer efficiency as well as a low Finn’s cycling index (0.224%). The system is found as an immature system with moderate ascendancy and system reserve (overhead). Moreover, concerning the ecologically sustainable production from this wetland more virtual mass-balanced models were developed (i) to evaluate the optimum grass carp stocking to manage the macrophyte population and (ii) to assess the impact of the Eichhornia on the system functioning. The results show that the system can harbour 1.5 times more grass carp biomass than the present without hampering sustainability. Concerning the impact of Eichhornia, it has been noticed that this macrophyte affects the maturity and transfer process of the system. Moreover, its huge biomass only accumulates more detritus in the ecosystem.

A nanozyme is a nanoscale material having enzyme-like properties. It exhibits several superior properties, including low preparation cost, robust catalytic activity, and long-term storage at ambient temperatures. Moreover, high stability enables repetitive use in multiple catalytic reactions. Hence, it is considered a potential replacement for natural enzymes. Enormous research interest in nanozymes in the past two decades has made it imperative to look for better enzyme-mimicking materials for biomedical applications. Given this, research on metal–organic frameworks (MOFs) as a potential nanozyme material has gained momentum. MOFs are advanced hybrid materials made of inorganic metal ions and organic ligands. Their distinct composition, adaptable pore size, structural diversity, and ease in the tunability of physicochemical properties enable MOFs to mimic enzyme-like activities and act as promising nanozyme candidates. This review aims to discuss recent advances in the development of MOF-based nanozymes (MOF-NZs) and highlight their applications in the field of biomedicine. Firstly, different enzyme-mimetic activities exhibited by MOFs are discussed, and insights are given into various strategies to achieve them. Modification and functionalization strategies are deliberated to obtain MOF-NZs with enhanced catalytic activity. Subsequently, applications of MOF-NZs in the biosensing and therapeutics domain are discussed. Finally, the review is concluded by giving insights into the challenges encountered with MOF-NZs and possible directions to overcome them in the future. With this review, we aim to encourage consolidated efforts across enzyme engineering, nanotechnology, materials science, and biomedicine disciplines to inspire exciting innovations in this emerging yet promising field.

Abstract It is undeniable that plastics are ubiquitous and a threat to global ecosystems. Plastic waste is transformed into microplastics (MPs) through physical and chemical disruption processes within the aquatic environment. MPs are detected in almost every environment due to their worldwide transportability through ocean currents or wind, which allows them to reach even the most remote regions of our planet. MPs colonized by biofilm-forming microbial communities are known as the ‘‘plastisphere”. The revelation that this unique substrate can aid microbial dispersal has piqued interest in the ground of microbial ecology. MPs have synergetic effects on the development, transportation, persistence, and ecology of microorganisms. This review summarizes the studies of plastisphere in recent years and the microbial community assemblage (viz. autotrophs, heterotrophs, predators, and pathogens). We also discussed plastic-microbe interactions and the potential sources of plastic degrading microorganisms. Finally, it also focuses on current technologies used to characterize those microbial inhabitants and recommendations for further research.

The global rise in antibiotic resistance, fueled by indiscriminate antibiotic usage in medicine, aquaculture, agriculture, and the food industry, presents a significant public health challenge. Urban wastewater and sewage treatment plants have become key sources of antibiotic resistance proliferation. The present study focuses on the river Ganges in India, which is heavily impacted by human activities and serves as a potential hotspot for the spread of antibiotic resistance. We conducted a metagenomic analysis of sediment samples from six distinct locations along the river to assess the prevalence and diversity of antibiotic resistance genes (ARGs) within the microbial ecosystem. The metagenomic analysis revealed the predominance of Proteobacteria across regions of the river Ganges. The antimicrobial resistance (AMR) genes and virulence factors were determined by various databases. In addition to this, KEGG and COG analysis revealed important pathways related to AMR. The outcomes highlight noticeable regional differences in the prevalence of AMR genes. The findings suggest that enhancing health and sanitation infrastructure could play a crucial role in mitigating the global impact of AMR. This research contributes vital insights into the environmental aspects of antibiotic resistance, highlighting the importance of targeted public health interventions in the fight against AMR.

Edwardsiella tarda is one of the serious bacterial pathogens infecting both cultured and wild catfish urging an immediate need for effective protection strategies. This study assessed the effects of dietary supplementation of Pseudomonas aeruginosa FARP72 at 108 cells/g feed (PA diet) for 30 days on the innate immunity parameters, viz., respiratory oxidative burst (ROB) activity, lysozyme, ceruloplasmin, myeloperoxidase, in-vitro nitric oxide (NO) production in addition to the expression of immune genes encoding interleukin-1β, C3 and transferrin in yellowtail catfish Pangasius pangasius and their resistance to Edwardsiella tarda challenge at a sub-lethal dose of 1.50 × 107 cells/fish. A significant increase in the innate immunity parameters was noted in PA diet-fed catfish on 30 dpf compared to the control. Post E. tarda challenge, the levels of immune parameters increased significantly and peaked at 5 dpi irrespective of feeding to confer protection against E. tarda. Their levels, however, decreased on and from 10 dpi. The results on the expression of immune genes encoding interleukin-1β, C3 and transferrin in the kidney and liver tissue samples of PA diet-fed P. pangasius upon challenge with E. tarda further confirmed the ability of P. aeruginosa to stimulate primary immune organs at the gene level. The effects of feeding P. aeruginosa FARP72 on the immune functions of catfish as examined by the functional immune assays, thus, demonstrating the innate immune responses of catfish that are differentially stimulated by the PA diet. The findings of our study would help evolve management strategies to confer protection against E. tarda infection in commercial catfish aquaculture.

Fisheries and aquaculture are the fastest-growing food-producing sector and rapidly becoming an important element for the global food security since they are the primary source of seafood and high animal protein in the human diet. Genome editing offers new possibilities such as the clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein (Cas9) technology, which has the potential to accelerate the sustainable genetic improvement in fisheries and aquaculture. The CRISPR/Cas9 system has four key components, namely, target DNA, Cas9, the protospacer adjacent motif sequence, and the guide RNA or single-guide RNA. CRISPR/Cas is cheaper, easier, and more precise than the other genome editing technologies and can be used as a new breeding technology in fisheries and aquaculture to solve the far-reaching challenges. The attributes like high fecundity, external fertilization, short generation interval, the established method of breeding, and the larval rearing of most aquaculture species have advantages for CRISPR/Cas9 genome editing applications. CRISPR/Cas9 has recently been applied to the traits valued in some aquaculture species (almost >20 species), targeting the main traits of traditional genetic improvement initiatives like growth, disease resistance, reproduction, sterility, and pigmentation. Genome editing can fast forward the breeding process with precision where changes occur in the targeted genes. The probability of desired changes occurring and passing the trait in the next generation is high, so it takes 1-3 generations to establish a breed. Moreover, CRISPR/Cas genome editing rapidly introduces favorable changes by disrupting genes with targeted minor changes, in contrast to transgenesis, which introduces foreign genes into the host genome and thereby alleviates major public concerns on safety. Although the CRISPR/Cas technology has a tremendous potential, there are several technical challenges and regulatory and public issues concerning the applications in fisheries and the aquaculture breeding sector. Nonetheless, the exciting point in the CRISPR/Cas9 genome editing is that two CRISPR-edited fish, namely, red sea bream and tiger puffer developed by the Kyoto-based startup got approval and are now on the market for sale, and another fish, FLT-01 Nile tilapia developed by the AquaBounty, is not classified under genetically modified organism regulatory. However, there is still a way to go before it revolutionizes and becomes viable in commercial aquaculture as the new breeding technology for aquaculture-important traits and species.

A case of severe mortality in farmed Labeo rohita was investigated to characterize the causative agent. We identified the bacterial strain as Aeromonas veronii isolated from the gut of infected L. rohita by biochemical assay, scanning electron microscopy and 16S rRNA gene sequence analysis. The in vivo challenge experiment showed that the LD50 of A. veronii was 2.2 × 104 CFU/fish. Virulence gene investigation revealed that the isolated A. veronii possesses Aerolysin, Cytotoxic enterotoxin, Serine protease, Dnase and Type III secretion system genes. The isolated strain was resistant to two antibiotics (ampicillin and dicloxacillin) while susceptible to 22 other antibiotics. The study further revealed that A. veronii induced both stresses along with non-specific and specific immune responses marked by elevated cortisol HSP70, HSP90 and IgM levels in the treated L. rohita fingerlings. Although the bacterial pathogen enhances the immune response, the negative effect on fish, including stress, and high mortality, create concern and a need for A. veronii management in L. rohita farms. The knowledge gained from this study would facilitate future research aimed at assessing the pathogenicity of A. veronii, with an emphasis on microbial disease management in other farmed fish species.

Acetylcholinesterase (AChE), a widely used enzyme for inhibition-based biosensors in pesticide residues detection, lags due to multiple-step operation, time-consuming incubation and reactivation/regeneration steps. Herein, this endeavour reports the development of Organophosphate Hydrolase (OPH), which has functional superiority over the AChE and explored in on-spot biosensing device for organophosphate pesticide residue detection in fruits and vegetables. The organophosphate degrading enzyme OPH is expressed from the ‘opd’ gene through biotechnological tools. The OPH exhibited its best activity at pH 8.0 and subsequently thermal inactivation over 37 °C. The activity of the purified OPH enzyme was found 2.75 U mL−1 at λmax 410 nm. Furthermore, the developed OPH is integrated into 96 well plate format with our previously reported UIISScan 1.1, an advanced imaging array technology based field-portable high-throughput sensory system. The developed biosensor revealed a linear range from 100 ng mL−1 to 0.1 ng mL−1 for detection of organophosphate pesticide residues with a negative slope i.e. y = 235.678x (ng mL−1) – 62.8725 with R2 = 0.99991 and n = 23. Moreover, the applicability of the developed biosensor was tested for market available fruits and vegetables. This is the first-ever reported OPH mediated on-spot biosensing device for pesticide residue detection in fruits and vegetables to the best of our knowledge.

Aphanomyces invadans is an aquatic oomycete pathogen and the causative agent of epizootic ulcerative syndrome (EUS) in fresh and brackish water fish, which is responsible for severe mortalities and economic losses in aquaculture. Therefore, there is an urgent need to develop anti-infective strategies to control EUS. An Oomycetes, a fungus-like eukaryotic microorganism, and a susceptible species, i.e., Heteropneustes fossilis, are used to establish whether an Eclipta alba leaf extract is effective against the EUS-causing A. invadans. We found that treatment with methanolic leaf extract, at concentrations between 50–100 ppm (T4–T6), protects the H. fossilis fingerlings against A. invadans infection. These optimum concentrations induced anti-stress and antioxidative response in fish, marked by a significant decrease in cortisol and elevated levels of superoxide dismutase (SOD) and catalase (CAT) levels in treated animals, as compared with the controls. We further demonstrated that the A. invadans-protective effect of methanolic leaf extract was caused by its immunomodulatory effect and is linked to the enhanced survival of fingerlings. The analysis of non-specific and specific immune factors confirms that methanolic leaf extract-induced HSP70, HSP90 and IgM levels mediate the survival of H. fossilis fingerlings against A. invadans infection. Taken together, our study provides evidence that the generation of anti-stress and antioxidative responses, as well as humoral immunity, may play a role in protecting H. fossilis fingerlings against A. invadans infection. It is probable that E. alba methanolic leaf extract treatment might become part of a holistic strategy to control EUS in fish species.

The biofloc system has recently attracted great attention as a cost-effective, sustainable, and environmentally friendly technology and expected to contribute toward human food security (Zero Hunger SDG 2). It is also expected that this endeavor can be adopted widely because of its characteristics of zero water exchange and reduced artificial feeding features. In the biofloc system, the flocs which are generally formed by aggregation of heterotrophic microorganisms, serve as natural bioremediation candidates. These microbes effectively maintain water quality by utilizing the nutrient wastes, mostly originated from digested, unconsumed, and metabolic processes of feed. Additionally, the flocs are important sources of nutrients, mainly a protein source, and when these are consumed by aquaculture animals they improve the growth performance, immunity, and disease tolerance of host against pathogenic microbial infection. Here in this review, we focus on recent advances that could provide a mechanistic insight on how the microbial community developed in the biofloc system helps in the bioremediation process and enhances the overall health of the host. We have also tried to address the possible role of these microbial communities against growth and virulence of pathogenic microbes.

Heat shock proteins (Hsps) are a family of ubiquitously expressed stress proteins and extrinsic chaperones that are required for viability and cell growth in all living organisms. These proteins are highly conserved and produced in all cellular organisms when exposed to stress. Hsps play a significant role in protein synthesis and homeostasis, as well as in the maintenance of overall health in crustaceans against various internal and external environmental stresses. Recent reports have suggested that enhancing in vivo Hsp levels via non-lethal heat shock, exogenous Hsps, or plant-based compounds, could be a promising strategy used to develop protective immunity in crustaceans against both abiotic and biotic stresses. Hence, Hsps as the agent of being an immune booster and increasing disease resistance will present a significant advancement in reducing stressful conditions in the aquaculture system.

The toll-like receptor 5 (TLR5) is the most conserved important pattern recognition receptors (PRRs) often stimulated by bacterial flagellins and plays a major role in the first-line defense against invading pathogenic bacteria and in immune homeostasis. Experimental crystallographic studies have shown that the extracellular domain (ECD) of TLR5 recognizes flagellin of bacteria and functions as a homodimer in model organism zebrafish. However, no structural information is available on TLR5 functionality in the major carp Cirrhinus mrigala (mrigala) and its interaction with bacterial flagellins. Therefore, the present study was undertaken to unravel the structural basis of TLR5-flagellin recognition in mrigala using structural homodimeric TLR5-flagellin complex of zebrafish as reference. Integrative structural modeling and molecular dynamics simulations were employed to explore the structural and mechanistic details of TLR5 recognition. Results from structural snapshots of MD simulation revealed that TLR5 consistently formed close interactions with the three helices of the D1 domain in flagellin on its lateral side mediated by several conserved amino acids. Results from the intermolecular contact analysis perfectly substantiate with the findings of per residue-free energy decomposition analysis. The differential recognition mediated by flagellin to TLR5 in mrigala involves charged residues at the interface of binding as compared to the zebrafish complex. Overall our results shows TLR5 of mrigala involved in innate immunity specifically recognized a conserved site on flagellin which advocates the scientific community to explore host-specific differences in receptor activation.

In this study, we report the presence of a microbial community of bioremediation potential in terms of relative abundance and taxonomic biodiversity in sediment samples of river Ganga and Yamuna, India at nine different sites. Metagenomic libraries were constructed using TruSeq Nano DNA Library Prep Kit and sequenced on NextSeq 500 by Illumina Next Generation Sequencing (NGS) technology. Bioremediation bacteria belong to 45 genera with 92 species and fungi belong to 13 genera with 24 species have been classified using Kaiju taxonomical classification. The study revealed that Proteobacteria was the most dominant bacterial flora, followed by Actinobacteria, Firmicutes, and Deinococcus–Thermus. PCA analysis revealed that bioremediation bacteria viz. Streptomyces bikiniensis, Rhodococcus qingshengii, Bacillus aerophilus, Pseudomonas veronii, etc., were more dominant in highly polluted river stretch as compared to less polluted river stretch. Similarly, the relative abundance of bioremediation fungi viz. Phanerochaete chrysosporium and Rhizopus oryzae, etc., were significantly correlated with the polluted Kanpur stretch of river Ganga. Several protein domains, which play a pivotal role in bioremediation in the polluted environments, including urea ABC transporter, UrtA, UrtD, UrtE, zinc/cadmium/mercury/lead-transporting ATPase, etc., were identified using protein domain analysis. The protein domains involved in pesticide biodegradation viz. P450, short-chain dehydrogenases/reductases (SDR), etc., were also discovered in river sediment metagenomics data. This is the first report on the richness of bioremediation microbial communities in the Ganga and Yamuna riverine ecosystems, highlighting their importance in aquatic pollution management.

In the present study, we identified salt stress tolerant genes from the marine bacterium Staphylococcus sp. strain P-TSB-70 through transcriptome sequencing. In favour of whole-genome transcriptome profiling of Staphylococcus sp. strain P-TSB-70 (GenBank Accn. No. KP117091) which tolerated upto 20% NaCl stress, the strain was cultured in the laboratory condition with 20% NaCl stress. Transcriptome analyses were performed by SOLiD4.0 sequencing technology from which 10280 and 9612 transcripts for control and treated, respectively, were obtained. The coverage per base (CPB) statistics were analyzed for both the samples. Gene ontology (GO) analysis has been categorized at varied graph levels based on three primary ontology studies viz. cellular components, biological processes, and molecular functions. The KEGG analysis of the assembled transcripts using KAAS showed presumed components of metabolic pathways which perhaps implicated in diverse metabolic pathways responsible for salt tolerance viz. glycolysis/gluconeogenesis, oxidative phosphorylation, glutathione metabolism, etc. further involving in salt tolerance. Overall, 90 salt stress tolerant genes were identified as of 186 salt-related transcripts. Several genes have been found executing normally in the TCA cycle pathway, integral membrane proteins, generation of the osmoprotectants, enzymatic pathway associated with salt tolerance. Recognized genes fit diverse groups of salt stress genes viz. abc transporter, betaine, sodium antiporter, sodium symporter, trehalose, ectoine, and choline, that belong to different families of genes involved in the pathway of salt stress. The control sample of the bacterium showed elevated high proportion of transcript contigs (29%) while upto 20% salt stress treated sample of the bacterium showed a higher percentage of transcript contigs (31.28%). A total of 1,288 and 1,133 transcript contigs were measured entirely as novel transcript contigs in both control and treated samples, respectively. The structure and function of 10 significant salt stress tolerant genes of Staphylococcus sp. have been analyzed in this study. The information acquired in the present study possibly used to recognize and clone the salt stress tolerant genes and support in developing the salt stress-tolerant plant varieties to expand the agricultural productivity in the saline system.

Beneficial microbes are all around us and it remains to be seen, whether all diseases and disorders can be prevented or treated with beneficial microbes. In this study, the presence of various beneficial bacteria were identified from the sediments of Indian major Rivers Ganga and Yamuna from nine different sites using a metagenomic approach. The metagenome sequence analysis using the Kaiju Web server revealed the presence of 69 beneficial bacteria. Phylogenetic analysis among these bacterial species revealed that they were highly diverse. Relative abundance analysis of these bacterial species is highly correlated with different pollution levels among the sampling sites. The PCA analysis revealed that Lactobacillus spp. group of beneficial bacteria are more associated with sediment sampling sites, KAN-2 and ND-3; whereas Bacillus spp. are more associated with sites, FAR-2 and ND-2. This is the first report revealing the richness of beneficial bacteria in the Indian rivers, Ganga and Yamuna. The study might be useful in isolating different important beneficial microorganisms from these river sediments, for possible industrial applications.

Shrimp, as a high-protein animal food commodity, are one of the fastest growing food producing sectors in the world. It has emerged as a highly traded seafood product, currently exceeding 8 MT of high value. However, disease outbreaks, which are considered as the primary cause of production loss in shrimp farming, have moved to the forefront in recent years and brought socio-economic and environmental unsustainability to the shrimp aquaculture industry. Acute hepatopancreatic necrosis disease (AHPND), caused by Vibrio spp., is a relatively new farmed penaeid shrimp bacterial disease. The shrimp production in AHPND affected regions has dropped to ~60%, and the disease has caused a global loss of USD 43 billion to the shrimp farming industry. The conventional approaches, such as antibiotics and disinfectants, often applied for the mitigation or cure of AHPND, have had limited success. Additionally, their usage has been associated with alteration of host gut microbiota and immunity and development of antibiotic resistance in bacterial pathogens. For example, the Mexico AHPND-causing V. parahaemolyticus strain (13-306D/4 and 13-511/A1) were reported to carry tetB gene coding for tetracycline resistance gene, and V. campbellii from China was found to carry multiple antibiotic resistance genes. As a consequence, there is an urgent need to thoroughly understand the virulence mechanism of AHPND-causing Vibrio spp. and develop novel management strategies to control AHPND in shrimp aquaculture, that will be crucially important to ensure food security in the future and offer economic stability to farmers. In this review, the most important findings of AHPND are highlighted, discussed and put in perspective, and some directions for future research are presented.

Soil salinization has become a severe constraint for crop production world-wide which necessitated development or induced enhancement of salt stress tolerance in plant life to sustain production in saline lands. Recognition and prospecting of valuable stress tolerant genes from natural microbial resources of saline habitat is obscure to date. Therefore, the investigation was towards isolation and characterization of marine salt stress tolerant microbes along the East coast of India for revelation of effective salt stress tolerant genes. Salt stress tolerance was assessed from 98 bacterial isolates obtained from 28 water and soil samples. Among them, 35 isolates which failed to grow beyond 4% salt were discarded and remainder 63 isolates were selected for further functional analysis and only seven isolates recorded ≥8% NaCl stress tolerance. Phylogeny revealed that four isolates belong to Firmicutes and three isolates were members of Proteobacteria. Ribosomal Database Project Release-11 and SILVA SSU database based genotyping and taxonomic identity analysis confirmed that the higher (20%) salt stress tolerant bacteria were Staphylococcus sp., Enterococcus sp., Enterobacter sp. and Proteus sp. To investigate candidate, as well as, novel salt stress tolerant genes, the seven bacterial isolates would provide new horizon to focus on the recent developments of salinity stress tolerance. In addition, the findings evidently point out the diversity of salt stress tolerant marine bacteria in coastal Odisha and West Bengal, India.

The complete mitogenome of Osteobrama belangeri is described using Ion Torrent (PGM sequencer), which was 16,609 bp in size comprising 13 mRNAs, teo rRNA genes, 22 tRNAs, and 926 bp as D-Loop control region, in addition to gene order and organization, being similar to most of the other related Cypriniformes fish mitogenome of NCBI databases. The all 22 tRNAs were packed into a typical clover-leaf structure. In the present study, the mitogenome has 99% similarity to the complete mitogenome sequence of O. belangeri mitogenome details previously and also would be helpful in understanding the phylogenetics, population genetics, and evolution of family Cyprinidae fishes.

Nucleotide-binding oligomerization domain-containing protein 1 (NOD1) and NOD2 are cytosolic pattern recognition receptors playing pivotal roles in innate immune signaling. NOD1 and NOD2 recognize bacterial peptidoglycan derivatives iE-DAP and MDP, respectively and undergoes conformational alternation and ATP-dependent self-oligomerization of NACHT domain followed by downstream signaling. Lack of structural adequacy of NACHT domain confines our understanding about the NOD-mediated signaling mechanism. Here, we predicted the structure of NACHT domain of both NOD1 and NOD2 from model organism zebrafish (Danio rerio) using computational methods. Our study highlighted the differential ATP binding modes in NOD1 and NOD2. In NOD1, γ-phosphate of ATP faced toward the central nucleotide binding cavity like NLRC4, whereas in NOD2 the cavity was occupied by adenine moiety. The conserved 'Lysine' at Walker A formed hydrogen bonds (H-bonds) and Aspartic acid (Walker B) formed electrostatic interaction with ATP. At Sensor 1, Arg328 of NOD1 exhibited an H-bond with ATP, whereas corresponding Arg404 of NOD2 did not. 'Proline' of GxP motif (Pro386 of NOD1 and Pro464 of NOD2) interacted with adenine moiety and His511 at Sensor 2 of NOD1 interacted with γ-phosphate group of ATP. In contrast, His579 of NOD2 interacted with the adenine moiety having a relatively inverted orientation. Our findings are well supplemented with the molecular interaction of ATP with NLRC4, and consistent with mutagenesis data reported for human, which indicates evolutionary shared NOD signaling mechanism. Together, this study provides novel insights into ATP binding mechanism, and highlights the differential ATP binding modes in zebrafish NOD1 and NOD2.