Bioaccumulation of Potentially Toxic Elements in Commercially Important Food Fish Species from Lower Gangetic Stretch: Food Security and Human Health Risk Assessment.

Ganga river is the inhabitant of more than 190 fish species and important river system of India. Potentially toxic elements (PTEs) in the Gangetic riverine ecosystem are a hot environmental issue. A detailed evaluation of PTEs bioaccumulation in Gangetic fishes is required to safeguard human health. The present study investigated the bioaccumulation of PTEs (Cd, Co, Cr, Cu, Li, Ni, Pb, Se, Zn, and Mn) within 12 economic fish species (n = 72) collected from the lower Gangetic stretch. The mean concentrations of PTEs followed the order Zn > Cu > Mn > Ni > Se > Cr > Pb > Co ~ Li > Cd. Li and Se bioaccumulation were studied first time from Gangetic fishes. Results demonstrated that all the selected PTEs were below the maximum permissible limit recommended by reference standards except for Zn in L. catla and L. rohita. For all PTEs, the metal pollution index (MPI), hazard quotient (THQ), and hazard index (HI) were < 1, indicating that these PTEs do not pose a health risk to the public through the dietary intake of fish in this study area. All studied fish were acceptable in terms of carcinogenic risk (CR) from exposure to Cd, Cr, and Pb. Multivariate statistical analysis suggests that inter-correlated metals have similar dispersion properties and bioaccumulation homology within the body. This study provides a scientific basis for food safety assessment and continuous monitoring of PTEs in Gangetic fishes is suggested in the future to safeguard human health.

Riverine connectivity influences the phytoplankton ecology in the open floodplain wetland of the lower river Ganga.

The river Ganga has several floodplain wetlands that support its ecology and ecosystem. Phytoplankton is an important component of the aquatic ecosystem, which plays an important role as a bioindicator for the assessment of aquatic health. The present study was conducted between 2018 and 2019 to understand the seasonal variation in the phytoplankton diversity of the Charaganga wetland and, parallelly, in the river Ganga in Nabadweep, India. The study explains how riverine connectivity affects the structure of the algal community in the wetland ecosystem. In the study, it has been observed that in the wetland, maximum mean phytoplankton density was noticed during pre-monsoon, i.e., 4079 unit l-1 followed by post-monsoon 3812 unit l-1 and monsoon 550 unit l-1, respectively. In the river system, the phytoplankton density varied from 78 unit l-1 to 653 unit l-1 seasonally, i.e., highest during monsoon and lowest during pre-monsoon. In both the ecosystems, i.e., wetland and river, the supreme influential group was Cyanophyceae followed by diatoms. One-way ANOVA showed a significant variation (p > 0.05) of three algal groups of phytoplankton (Bacillariophyceae, Coscinodiscophyceae, Chlorophyceae) in the river, while in the wetland, no significant variation (p > 0.05) was found among the other algal groups. The observed higher Shannon and Margalef's species richness value in the wetland was observed than in the river defines the significance and importance of the wetland ecosystem, which may support the growth and conservation of various aquatic organisms as well. The study highlighted that the influencing abiotic factors like water temperature, dissolved oxygen, pH, and nutrients have affected the phytoplankton community in both the water bodies, i.e., wetland and river. We concluded that river connectivity is required to restore the biotic flora of the wetland ecosystem.

Receptor model-based source apportionment and ecological risk assessment of metals in sediment of river Ganga, India.

Ganga river surface sediment was sampled from 11 locations, which revealed average concentrations (mg/kg) of metals in the order Mn (296.93) > Zn (61.94) > Cr (54.82) > Cu (30.19) > Pb (24.42) > Cd (0.36). Sediment quality guidelines showed metals rarely to occasionally exhibit adverse biological effects. Indices like potential ecological risk, contamination security index, hazard quotients, multiple probable effect concentrations quality, mean probable effects level quotients, mean effects range median quotient suggest nil to a very low level of pollution with low ecological risk. Contamination factor, geo accumulation index, enrichment factor, quantification of contamination revealed that Pb and Cd originated from anthropogenic activities. APCS-MLR model revealed that metals contributed from natural sources (Zn, Mn, Cr; 20.18 %), industrial-agricultural (Cd; 21.35 %); and discharge of paints, Pb batteries, fossil fuel (Pb; 8.49 %). Present findings will serve as an effective guideline for managing and ameliorating pollution in the river system.

Microbial inoculums improve growth and health of Heteropneustes fossilis via biofloc-driven aquaculture.

Biofloc technology aims to maximize fish farming productivity by effectively breaking down ammonia and nitrite, promoting healthy flocculation, and enhancing the growth and immunity of cultured animals. However, a major limitation in this field is the suitable starter microbial culture and narrow number of fish species that have been tested with the biofloc system. Here, we investigated various microbial inoculum containing beneficial microbes with probiotics, immunostimulatory and flocs development and bioremediation properties would lead to the development of ideal biofloc development. Three treatment groups with different microbial combinations, viz., group 1 [Bacillus subtilis (AN1) + Pseudomonas putida (PB3) + Saccharomyces cerevisiae (ATCC-2601)], group 2 [B. subtilis (AN2) + P. fluorescens (PC3) + S. cerevisiae (ATCC-2601)] and group 3 [B. subtilis (AN3) + P. aeruginosa (PA2) + S. cerevisiae (ATCC-2601)] were used and compared with the positive control (pond water without microbial inoculums) and negative control (clear water: without microbial inoculums and carbon sources) on biofloc development and its characteristic features to improve the water quality and growth of fish. We demonstrated that microbial inoculums, especially group 2, significantly improve the water quality and microbiota of flocs and gut of the test animal, Heteropneustes fossilis. The study further demonstrates that biofloc system supplemented with microbial inoculums positively regulates gut histomorphology and growth performance, as evidenced by improved villous morphology, amylase, protease and lipase activity, weight gain, FCR, T3, T4 and IGF1 levels. The inoculums induced an antioxidative response marked by significantly higher values of catalase (CAT) and superoxide dismutase (SOD) activity. Furthermore, the supplementation of microbial inoculums enhances both specific and non-specific immune responses and significantly elevated levels of immune genes (transferrin, interleukin-1ß and C3), and IgM was recorded. This study provides a proof-of-concept approach for assessing microbial inoculums on fish species that can be further utilized to develop biofloc technology for use in sustainable aquaculture.

Microbiome analysis reveals potential for modulation of gut microbiota through polysaccharide-based prebiotic feeding in Oreochromis niloticus (Linnaeus, 1758).

Characterization and functional profiling of the gut microbiota are essential for guiding nutritional interventions in fish and achieving favorable host-microbe interactions. Thus, we conducted a 30 days study to explore and document the gut microbial community of O. niloticus, as well as to evaluate the effects of a polysaccharide-based prebiotics with 0.5% and 0.75% Aloe vera extract on the gut microbiome through genomic analysis. The V3-V4 region of 16S rRNA was amplified and sequenced using Illumina HiSeq 2500, resulting in 1,000,199 reads for operational taxonomic unit (OTU) identification. Out of 8,894 OTUs, 1,181 were selected for further analysis. Our results revealed that Planctomycetes, Firmicutes, Proteobacteria, Verrucomicrobia, Actinobacteria, and Fusobacteria were the dominant phyla in both control and treatment samples. Higher doses of prebiotics were found to improve Planctomycetes and Firmicutes while decreasing Proteobacteria and Verrucomicrobia. We observed increasing trends in the abundance of Bacilli, Bacillaceae, and Bacillus bacteria at the class, family, and genus levels, respectively, in a dose-dependent manner. These findings were consistent with the conventional colony count data, which showed a higher prevalence of Bacillus in prebiotic-supplemented groups. Moreover, predicted functional analysis using PICRUSt indicated a dose-dependent upregulation in glycolysis V, superpathway of glycol metabolism and degradation, glucose and xylose degradation, glycolysis II, and sulfoglycolysis pathways. Most of the energy, protein, and amino acid synthesis pathways were upregulated only at lower doses of prebiotic treatment. Our findings suggest that the gut microbiome of O. niloticus can be optimized through nutritional interventions with plant-based polysaccharides for improved growth performance in commercial fish.

Aeromonas veronii Is a Lethal Pathogen Isolated from Gut of Infected Labeo rohita: Molecular Insight to Understand the Bacterial Virulence and Its Induced Host Immunity.

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.

Hemato-biochemical alteration in the bronze featherback Notopterus notopterus (Pallas, 1769) as a biomonitoring tool to assess riverine pollution and ecology: a case study from the middle and lower stretch of river Ganga.

Fishes are poikilothermic animals and are rapid responders to any sort of ecological alteration. The responses in the fish can be easily assessed from their hematological and biochemical responses. To study the variation in the hemato-biochemical parameters in retort to ecological alteration and ecological regime, a study was conducted at six different sampling stations of the middle and lower stretches of river Ganga. Various hematological and biochemical responses of fishes were also monitored in response to multiple ecological alterations. For the assessment of ecological alteration, various indices were calculated such as the water pollution index (WPI), National Sanitation Foundation-water quality index (NSF-WQI), and Nemerow's pollution index (NPI) has been calculated based on various water quality parameters such as dissolved oxygen (DO), pH, total dissolved solids (TDS), total alkalinity (TA), total hardness (TH), electrical conductivity (EC), biochemical oxygen demand (BOD), chlorinity (CL), total nitrogen (TN), and total phosphorus (TP). The hematological parameters such as WBC, RBC, platelet, hemoglobin, and hematocrit were monitored. The serum biochemical parameters such as SGPT, SGOT, ALP, amylase, bilirubin, glucose, triglyceride (TRIG), and cholesterol (CHOL) were investigated. The study revealed that NSF-WQI varied from 45.08 at Buxar to 110.63 at Rejinagar and showed a significantly positive correlation with SGPT, SGOT, ALP, TRIG, CHOL, and WBC, whereas a significantly negative correlation was observed between TRIG and RBC. WPI varied from 19 to 23 and showed a significant positive correlation with SGOT and a negative correlation was observed with total nitrogen. The PCA analysis illustrated the significance of both natural as well as anthropogenic factors on riverine ecology. Strong positive loading was observed with SGPT, SGOT, ALP, and platelet. The study signified the need for monitoring the hemato-biochemical responses of fishes in response to alterations in the ecological regime.

Immunomodulatory Potency of Eclipta alba (Bhringaraj) Leaf Extract in Heteropneustes fossilis against Oomycete Pathogen, Aphanomyces invadans.

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.

Integrated biomarker approach strongly explaining in vivo sub-lethal acute toxicity of butachlor on Labeo rohita.

Butachlor herbicide belongs to the family of chloroacetanilide group, widely used for control of grass and broadleaf weeds in paddy fields however, its repeated application may result in aquatic pollution. Butachlor residue has been detected in aquatic environments, which may produce toxic effects on non-target organisms including fish. Keeping this in mind, the present study was designed to estimate the LC50 of butachlor (Shaktiman®), and to evaluate the sub-lethal toxicity at two concentrations (12.42 µg L-1 and 62.10 µg L-1) in Labeo rohita for a period of 24, 48, and 72 h. Fish exposed to butachlor reduced the counts of red blood cells (RBC), haemoglobin (HGB), hematocrit (HCT), and white blood cells (WBC). A significant (p < 0.05) increase in the antioxidant enzyme (superoxide dismutase-SOD, glutathione-s-transferase-GST), and hepatic enzyme (glutamate-oxaloacetate transaminase-GOT, glutamate-pyruvate transaminase-GPT) were noticed in butachlor exposed fish. Heat shock protein 70 (HSP70) and HSP90 in gill; cortisol, protein, albumin, globulin, and triglyceride in serum were increased upon exposure of butachlor. On the contrary, complement 3 (C3) and immunoglobulin (IgM) in serum was found to be decreased compared to control fish. The findings thus suggest that the fish upon exposure to butachlor disrupts the biomarkers which ultimately leads to growth retardation in fish.

Multidecadal assessment of environmental variables in the river Ganga for pollution monitoring and sustainable management.

The Ganga River is the major source of drinking water for humans over the decades. It is also the ecological niche for millions of relict species, i.e., for a variety of planktons, benthic organisms, fish, and various other aquatic organisms. The blasting population resulted in an enhanced rate of pollution in the river system emanating from various anthropogenic activities and industrialization in the bank of river Ganga. The study was made in the middle and lower stretch of the river to monitor the decadal changes in the water quality of river Ganga from 1960 to 2019 at six different study sites. In the present study, various water quality parameters such as dissolved oxygen, pH, free carbon dioxide, total alkalinity, conductivity, total dissolved solids (TDS), hardness, chloride, and nitrate have been studied during 2015-2019. The data for 1960 to 2006 were taken from ICAR-CIFRI publications. Based on the studied parameters, National Sanitation Foundation (NSF)-water quality index (WQI) was calculated. In the present study, it was found that the calculated NSF-WQI was 69.24 in 1960-1961 which increased up to 113.39 during 2001-2006. But, with the implementation of various rejuvenating strategies, the WQI of the river got reduced to 106.48 during 2015-2019. This reflected the positive changes in the riverine system. Different water quality parameters such as dissolved oxygen, pH, and hardness were observed mostly within the permissible range as based on the drinking water guidelines for humans and survival of the aquatic organisms as well, except a few location-specific observations.

Water quality assessment in the ecologically stressed lower and estuarine stretches of river Ganga using multivariate statistical tool.

Water quality of the Ganga River system is changing day by day due to multifold increase in population, especially near the banks of river Ganga, and associated exponential amplification of anthropogenic activities also played a remarkable role in it. The ecologically important lower and estuarine stretch of river Ganga comprising 7 different sampling stations, i.e., Jangipur, Berhampore, Balagarh, Tribeni, Godakhali, Diamond Harbour and Fraserganj, were selected for the study as the stretch is enriched with the vast number of floral and faunal diversity. The study was conducted for a period of 5 years, i.e., from 2016 to 2020. In the study, various analytical tools and techniques were used for the assessment of riverine water quality, i.e., for calculation of water quality index (WQI); The National Sanitation Foundation Water Quality Index (NSF-WQI) and the Canadian Council of Ministers of the Environment Water Quality Index (CCME-WQI) were used for the assessment. Along with WQI various statistical univariate as well as multivariate analytical tools like principal component analysis, correlation, ANOVA, and cluster analysis were also used to achieve the desired outputs. In the study, it has been observed that NSF-WQI varied from 61 to 2552, in which the higher value of NSF-WQI denoted the unsuitability of the water quality concerning the drinking water standards and vice versa. The CCME-WQI represented a similar trend as that of NSF-WQI, as it varied from 18 to 92 in which the lower value denoted degradation in the drinking water quality and vice versa. The study revealed that the Diamond Harbour-Fraserganj stretch is having an undesired level of water quality which were analyzed based on the drinking water guideline values of the Bureau of Indian Standards and that of NSF-WQI and CCME-WQI.

Stocking density mediated stress modulates growth attributes in cage reared Labeo rohita (Hamilton) using multifarious biomarker approach.

The present study was conducted for 240 days to evaluate the effects of stocking density based on growth attributes, digestive enzymes, muscular composition, biochemical and physiological responses of Labeo rohita fingerlings in tropical inland open water cages. L. rohita (30.35 ± 1.08 g) were randomly distributed into three treatments, namely low stocking density, LSD (10 m-3), medium stocking density, MSD (20 m-3) and high stocking density, HSD (30 m-3) in triplicates. Fish were fed twice daily with CIFRI CAGEGROW® floating feed (crude protein-28%, crude fat-4%). Fish growth and feed efficiency were higher (p < 0.05) in LSD, however, MSD registered a higher yield. Amylase and protease activity reduced whereas lipase activity increased with increasing stocking density. Muscle crude protein and crude fat formed an inverse correlation. The fillet quality deteriorated at higher stocking densities based on Muscle pH, drip loss and frozen leakage rate. The stress biomarkers level (glucose, cortisol, superoxide dismutase and catalase) increased in serum under crowding conditions. Glutamate oxaloacetate transaminase and glutamate pyruvate transaminase in serum were significantly increased in HSD. Serum protein levels decreased with the increase in stocking densities. Body ionic imbalance (Na+, Cl- and K+) was observed under crowding stress. Based on growth attributes and multiple biomarker responses, L. rohita @ 10 m-3 was found to be the optimum density for inland open water cage culture.

Assessment of the effect of sub-lethal acute toxicity of Emamectin benzoate in Labeo rohita using multiple biomarker approach.

Emamectin benzoate (EMB) is a potent neurotoxin agent, widely used for ectoparasites control in aquaculture, but their detailed toxicological implications in Labeo rohita are unknown. Thus, this study was conceptualized to determine the LC50 and to investigate the effects of two sub-lethal concentrations 1/50th of 96 h LC50 (1.82 µgL-1) and 1/10thof 96 h LC50 (9.1 µgL-1) on hemato-immunological and biochemical responses in L. rohita (mean weight 25.54 ± 2.3 g and length 10.35 ± 2.4 cm) for a period of 24 h, 48 h, and 72 h. LC50 of EMB were 163 µgL-1, 112 µgL-1, 99 µgL-1 and 91 µgL-1 at 24 h, 48 h, 72 h, and 96 h respectively. The safe limit at 96 h LC50 of EMB was 2.30 µgL-1. In EMB treated fish, red blood cells, white blood cells, hemoglobin, and hematocrit counts were reduced (p < 0.05) significantly. Superoxide dismutase (SOD) activity in the liver and kidney declined (p < 0.05) at 72 h while in gill and muscle the activity increased significantly. Glutathione-s-transferase (GST) activity in the liver, gill, and kidney increased (p < 0.05) while muscle decreased significantly. Catalase (CAT) activity in liver, gill, and muscle decreased while in kidney increases. Glutamic-oxaloacetic acid transaminase (GOT) activity and Glutamate pyruvate transaminase (GPT) activity were increased in liver, kidney, and muscle tissue. The change in serum triglycerides, serum protein level was noticed. The level of cortisol, heat shock protein 70 (HSP70), and HSP90 increased (p < 0.05) while the immunological responses like immunoglobulin M (IgM) and complement 3(C3) activity decreased (p < 0.05) in EMB exposed fish. Thus, EMB exposure at two sub-lethal concentrations in L. rohita induces several hemato-immuno, and biochemical alterations in blood, serum, and different organs. The overall result of the present study indicated that EMB is toxic to fish even for a short-term exposure and low doses, and therefore utmost caution should be taken to prevent their drainage into water bodies.

Variation of Aulacoseira granulata as an eco-pollution indicator in subtropical large river Ganga in India: a multivariate analytical approach.

Aulacoseira granulata (Ehrenberg) Simonsen 1979 are considered as the eco-variable species which varies in density and diversity along with their morphological traits with the interference of environmental changes, so it is considered as one of the major ecological indicators of the water quality of lotic as well as lentic aquatic ecosystems. To assess major environmental factors which contribute to A. granulata bloom in the riverine system, a study was carried out from 2018 to 2019 comprising four different seasons at 11 sampling sites of river Ganga in the middle and lower stretch of river Ganga comprising freshwater and estuarine zones. For the analysis, different univariate, as well as multivariate, analytical tools such as principal component analysis (PCA) and water pollution index (WPI) were used. In the finding, it was observed that the average abundance of A. granulata was found maximum during the winter season. Among all the studied sites, the maximum average abundance was at Balagarh (71,576 cell l-1) and minimum at Diamond Harbour (68 cell l-1). The environmental factors such as dissolved oxygen, depth, and altitude showed a significant influence on the growth of A. granulata, while the water temperature negatively influenced the growth rate of A. granulata. The WPI showed a significantly negative correlation with cell length. Finally, the study concludes that the blooming of A. granulata is highly influenced by varied environmental conditions along the river Ganga, suggesting possible eutrophication. Therefore, a certain minimum flow and depth especially during the lean season have to be maintained for the sustenance of planktonic biota in the river Ganga.

Biofloc Microbiome With Bioremediation and Health Benefits.

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.

Elucidating the molecular interaction of Zebrafish (Danio rerio) peptidoglycan recognition protein 2 with diaminopimelic acid and lysine type peptidoglycans using in silico approaches.

Peptidoglycan recognition proteins (PGRPs) belong to the family of pattern recognition receptor, represent the major constituent of innate immunity. Although PGRPs are structurally conserved through evolution, their involvement in innate immunity is different in vertebrates and invertebrates. They are highly specific towards recognition of ligands and can hydrolyze bacterial peptidoglycans (PGNs). Zebrafish PGRPs (zPGRPs) have both peptidoglycans lytic amidase activity and broad-spectrum bactericidal activity, but far less is known about how these receptors recognize these microbial ligands. Such studies are hindered due to lack of structural and functional configuration of zPGRPs. Therefore, in this study, we predicted the three-dimensional structure of zPGRP2 through theoretical modeling, investigated the conformational and dynamic properties through molecular dynamics simulations. Molecular docking study revealed the microbial ligands, that is, muramyl pentapeptide-DAP , muramyl pentapeptide-LYS, muramyl tripeptide-DAP, muramyl tripeptide-Lys, muramyl tetrapeptide-DAP, muramyl tetrapeptide-LYS and tracheal cytotoxin interacts with the conserved amino acids of the ligand recognition site comprised of ß1, α2, α4, ß4 and loops connecting ß1 - α2, α2 - ß2, ß3 - ß4 and α4 - α5. Conserved His31, His32, Ala34, Ile35, Pro36, Lys38, Asp60, Trp61, Trp63, Ala89, His90, Asp106, His143 and Arg144 are predicted to essential for binding and provides stability to these zPGRP-PGN complexes. Our study provides basic molecular information for further research on the immune mechanisms of PGRP's in Zebrafish. The plasticity of the zPGRP's binding site revealed by these microbial ligands suggests an intrinsic capacity of the innate immune system to rapidly evolve specificities to meet new microbial challenges in the future.Communicated by Ramaswamy H. Sarma.