Biomarkers In Fish Research Articles

Identifying Universal Fish Biomarker Genes in Response to PCB126 Exposure by Comparative Transcriptomic Analyses.

Water pollution remains a major environmental concern, with increased toxic by-products being released into water bodies. Many of these chemical contaminants persist in the environment and bio-accumulate in aquatic organisms. At present, toxicological tests are mostly based on laboratory tests, and effective methods for monitoring wild aquatic environments remain lacking. In the present study, we used a well-characterized toxic chemical, 3,3',4,4',5-polychlorinated biphenyl (PCB126), as an example to try to identify common biomarker genes to be used for predictive toxicity of this toxic substance. First, we used two laboratory fish models, the zebrafish (Danio rerio) and medaka (Oryzias latipes), to expose PCB126 to obtain liver transcriptomic data by RNA-seq. Comparative transcriptomic analyses indicated generally conserved and concerted changes from the two species, thus validating the transcriptomic data for biomarker gene selection. Based on the common up- and downregulated genes in the two species, we selected nine biomarker genes to further test in other fish species. The first validation experiment was carried out using the third fish species, Mozambique tilapia (Oreochromis mossambicus), and essentially, all these biomarker genes were validated for consistent responses with the two laboratory fish models. Finally, to develop universal PCR primers suitable for potentially all teleost fish species, we designed degenerate primers and tested them in the three fish species as well as in another fish species without a genomic sequence available: guppy (Poecilia reticulata). We found all the biomarker genes showed consistent response to PCB126 exposure in at least 50% of the species. Thus, our study provides a promising strategy to identify common biomarker genes to be used for teleost fish analyses. By using degenerate PCR primers and analyzing multiple biomarker genes, it is possible to develop diagnostic PCR arrays to predict water contamination from any wild fish species sampled in different water bodies.

Environmental health in the upper-middle Luján River basin from a multi-biomarker approach

The objectives of this study were to evaluate the ecophysiological state of the biota using a set of biomarkers in the upper-middle Luján River. To this aim, we collected adult Cnesterodon decemmaculatus fish, biofilm and water at three sampling sites in the upper-middle Luján River (S1: rural area, S2: Luján City and S3: urban area after passing Lujan City). For each site we determined physicochemical variables, heavy metal concentration in water, 19 biomarkers in fish (morphometric, histological, genotoxic, oxidative stress, metabolic and neurotoxic) and six biomarkers in biofilm (oxidative stress and extracellular enzyme). Additionally, we compared the responses of fish and biofilm with those of laboratory controls obtained from outdoor cultures. Our results indicated increased heavy metal concentration at all sites, mainly As and Cd, and decreased dissolved oxygen at S1 and S3. In fish, genotoxic biomarkers showed significant differences with respect to the control. The comet assay indicated damage in fish at the urbanized sites (S2 and S3) and an increased frequency of erythrocytes with nuclear aberrations at all sites. The CEA index (cellular energy allocation), calculated from the metabolic biomarkers and lipid concentration were significantly increased at S1. The gill damage evaluated histologically and with three indices indicated severe damage at all sites. Gills showed thickened primary and secondary lamellae and fusion of filaments at all sites, but a significant increase in mucous cells was only found at S1 and S3. Biofilm showed increased values of extracellular enzymes (β-glucosidase and alkaline phosphatase, lipid peroxidation and oxidative stress enzymes (i.e., catalase) at S3. These results are novel in that they incorporated laboratory controls allowing for comparisons with fish and biofilm from the field. They provided information on the status of a fish population and biofilm community, indicating the negative effect of river water deterioration on the tested organisms. Moreover, results showed what biomarkers were most sensitive for each biological sample.

Metal ion-complexed DNA probe coupled with CRISPR/Cas12a amplification and AuNPs for sensitive colorimetric assay of metallothionein in fish

The accurate and sensitive detection of metallothionein (MT) is of great significance in the fields of biomedical, toxicological and environmental sciences. In this work, based on the high affinity interaction between MT and the heavy metal ions of Hg2+ and the significant signal amplification capability of Cas12a/crRNA enzyme as well, we report a simple and highly sensitive method for visual detection of MT, a biomarker in fish for heavy metal ion-induced water bio-pollution. The target MT molecules bind Hg2+ in the Hg2+- complexed hairpin DNA probes to unfold the hairpin structure into ssDNAs, which hybridize with the partial dsDNA duplexes via strand displacement to yield specific sequence-containing dsDNAs. Cas12a/crRNA recognizes these specific sequences to activate its enzyme activity to cyclically cleave the ssDNA linkers in the blue colored gold nanoparticle aggregates to transit their color into red to realize visual detection of MT. Owing to the signal amplification by Cas12a/crRNA, as low as 25 nM of MT can be visually detected with naked eye. In addition, our colorimetric detection method has high selectivity for MT against other interference proteins and can detect MT in the livers and kidneys of crucian carps bought from a local supermarket. Moreover, the developed assay overcomes the limitations of conventional MT detection methods in terms of complexity, high cost and low sensitivity and can therefore offer new methods for monitoring water bio-pollutions.

Calcium reduces the effects of cadmium on the responses of biomarkers in freshwater fish (Oreochromis niloticus)

ABSTRACT The hardness of freshwaters varies depending on geographical locations and calcium is the most important element determining water hardness. Thus, the present study focused on responses of oxidative stress biomarkers in the liver and ATPase activities in the gill of freshwater fish (Oreochromis niloticus) after acute and chronic cadmium exposures in differing calcium levels (30, 60 and 120 mg Ca/L). Fish were exposed to Cd for acute (25 µM, 3 days) and chronic (5 µM, 30 days) durations. There were significant (p < 0.05) alterations in the oxidative stress parameters after cadmium exposures. In the liver, activities of catalase and superoxide dismutase increased in both durations, though activities of glutathione peroxidase, glutathione reductase and glutathione S-transferase decreased. Likewise, levels of glutathione and malondialdehyde increased significantly in both durations. In the gill, there were significant decreases in Na,K-ATPase activity and increases in Mg-ATPase activity. However, calcium alone did not cause any alteration (p > 0.05) in the responses of biomarkers. Data also showed that alterations in biomarkers were highest at the lowest calcium levels. Present data emphasised that calcium protected the fish against the toxic effects of cadmium and suggested that metal toxicity varies depending on the chemical quality of waters from different geographical areas.

Multispecies and multibiomarker assessment of fish health from Iguaçu River reservoir, Southern Brazil.

This study investigated the impact of micropollutants on fish health from Segredo hydroelectric reservoir (HRS) along the Iguaçu River, Southern Brazil, contaminated by urban, industrial, and agricultural activities. This is the first comprehensive study assessment in the river after the severe drought in the 2020s in three fish species from different trophic levels Astyanax spp. (water column depth/omnivorous), Hypostomus commersoni (demersal/herbivorous), and Pimelodus maculatus (demersal/omnivorous). Animals, water, and sediment samples were collected from three distinct sites within the reservoir: Floresta (upstream), Iratim (middle), and Station (downstream). The chemical analysis revealed elevated concentrations of metals (Al, Cu, Fe) and the metalloid As in water, or Cu, Zn, and As in sediment, surpassing Brazilian regulatory limits, while the organic pollutants as DDT, PAHs, PCBs, and PBDEs were found under the Brazilian regulatory limits. The metal bioaccumulation was higher in gills with no significant differences among sites. The species Astyanax spp. and H. commersoni displayed variations in hepatosomatic index (HSI) and P. maculatus in the condition factor index (K) between sites, while adverse effects due to micropollutants bioaccumulation were observed by biochemical, genotoxic, and histopathological biomarkers. The principal component analysis and integrated biomarker response highlighted the upstream site Floresta as particularly inhospitable for biota, with distinctions based on trophic level. Consequently, this multifaceted approach, encompassing both fish biomarkers and chemical analyses, furnishes valuable insights into the potential toxic repercussions of micropollutant exposure. These findings offer crucial data for guiding management and conservation endeavors in the Iguaçu River.

Organic contamination and multi-biomarker assessment in watersheds of the southern Brazil: an integrated approach using fish from the Astyanax genus.

We aimed to examine the responses of pollution biomarkers in feral fish from Astyanax genus collected at three hydrographic regions in southern Brazil and the capacity of these tools to differentiate between various levels of contamination. To achieve this, levels of organochlorine pesticides (liver), as well as the biomarkers AChE (muscle and brain), TBARS (liver), and EROD (liver) were assessed. Collections were conducted in four municipalities (Alegrete, Caraá, Lavras, and Santa Vitória) during 1year, encompassing winter and summer. Fish from Alegrete were the most contaminated overall, but animals sampled in Caraá, and Lavras also displayed elevated levels of current-use pesticides. Elevated levels of endosulfans, DDTs, HCHs, and current-use pesticides were accompanied by elevated levels of TBARS in the liver. Conversely, fish from Santa Vitória exhibited the highest levels of PAHs, accompanied by elevated levels of EROD in the liver and reduced levels of AChE in muscle and brain. TBARS proved to be a reliable biomarker for assessing impacts arising from pesticide accumulation, while EROD and AChE served as valuable indicators of impacts resulting from PAHs accumulation. Ultimately, the results obtained in this study demonstrate the reliable use of the proposed biomarkers for tracking biological impacts stemming from aquatic pollution using feral Astyanax as biomonitoring species.

Assessing the water quality in a World Heritage Site using biomarkers in top fish predators

The use of biomarkers in fish for biomonitoring is a valuable approach to reveal effects of human impacts on biota health. Top predator fish are effective models for monitoring human activities' impacts on aquatic ecosystems. The Guaraguaçu River is the largest river-system on coastal region of South Brazil and a World Heritage site. The river receives contaminants from disorderly urban growth, including discharges of domestic sewage and small fishery boats, particularly during the tourist season. Our study aimed to assess impact of anthropogenic activities on water quality in the Guaraguaçu River by analyzing environmental contamination biomarkers in the top fish predator Hoplias malabaricus. Fish were collected using a fyke net trap across sectors representing a gradient of anthropic impact: sector 1 - pristine; sector 2 – impacted; and sector 3 - less impacted. Water samples were collected to analyze the presence of trace elements and pesticide. Biomarkers of the antioxidant system, histopathology, genotoxicity, neurotoxicity, and concentration of trace elements were analyzed in fish tissues. In water samples Al, Fe and Mn were detected, but no pesticides were found. In fish muscle, zinc and iron were detected. Brain acetylcholinesterase activity decreased in impacted sectors, indicating neurotoxic effects. The antioxidant system increased activity in gills and liver, and damage from lipoperoxidation was observed, particularly in sector 2 when compared to sector 1, suggesting oxidative stress. Histopathological biomarkers revealed lesions in the liver and gills of fish in impacted sectors. Micronuclei, a genotoxicity biomarker, were observed in organisms from all sectors. Our results demonstrate detrimental effects of poor water quality on biota health, even when contaminants are not detected in water.

Toxicological assessment of bromoxynil and 2-methyl-4-chlorophenoxyacetic acid herbicide in combination on Cirrhinus mrigala using multiple biomarker approach

The widespread application of herbicides raises concerns about their impact on non-target aquatic organisms. This study aimed to evaluate the toxicity of a commercially available herbicide formulation containing Bromoxynil+MCPA (2-Methyl-4-chlorophenoxyacetic acid) on Cirrhinus mrigala (economically significant fish). A total of 210 juvenile fish were subjected to a triplicate experimental setup, with 70 fish allocated to each replicate, exposed to seven different concentrations of herbicide: 0 mg/L, 0.133 mg/L, 0.266 mg/L, 0.4 mg/L, 0.5 mg/L, 0.66 mg/L, and 0.8 mg/L, respectively, for a duration of 96 h. The median lethal concentration (LC50) was determined to be 0.4 mg/L. Significant hematological alterations were observed, including decreases in RBC counts, hemoglobin, hematocrit, and lymphocyte counts, along with an increase in erythrocyte indices. Biochemical analysis revealed elevated levels of neutrophils, WBCs, bilirubin, urea, creatinine, ALT, AST, ALP, and glucose in treated groups. Morphological abnormalities in erythrocytes and histopathological changes in gills, liver, and kidneys were noted. Pathological alterations in gills, liver and kidneys including epithelial cell uplifting, lamellar fusion, hepatolysis, and renal tubule degeneration were observed. Oxidative stress biomarkers such as TBARS (Thiobarbituric Acid Reactive Substance), ROS (Reactive Oxygen Species), and POD (Peroxides) activity increased, while antioxidant enzymatic activities decreased as toxicant doses increased from low to high concentrations. The study reveals that Bromoxynil+MCPA significantly disrupts physiological and hematobiochemical parameters in Cirrhinus mrigala, which highlights the substantial aquatic risks. In conclusion, the herbicide formulation induced significant alterations in various fish biomarkers, emphasizing their pivotal role in assessing the environmental impact of toxicity. This multi-biomarker approach offers valuable insights regarding the toxicological effects, thereby contributing substantially to the comprehensive evaluation of environmental hazards.

Metabolomic and Lipidomic Tools for Tracing Fish Escapes from Aquaculture Facilities.

During adverse atmospheric events, enormous damage can occur at marine aquaculture facilities, as was the case during Storm Gloria in the southeastern Spanish Mediterranean in January 2020, with massive fish escapes. Fishes that escape were caught by professional fishermen. The objective of this study was to identify biomarkers in fish that enable differentiation among wild fish, escaped farm-raised fish, and farm-raised fish kept in aquaculture facilities until their slaughter. We focused on gilthead sea bream (Sparus aurata). We used nuclear magnetic resonance to search for possible biomarkers. We found that wild gilthead sea bream showed higher levels of taurine and trimethylamine-N-oxide (TMAO) in their muscle and higher levels of ω-3 fatty acids, whereas farm-escaped and farmed gilthead sea bream raised until slaughter exhibit higher levels of ω-6 fatty acids. From choline, carnitine, creatinine, betaine, or lecithin, trimethylamine (TMA) is synthesized in the intestine by the action of bacterial microflora. In the liver, TMA is oxidized to TMAO and transported to muscle cells. The identified biomarkers will improve the traceability of gilthead sea bream by distinguishing wild specimens from those raised in aquaculture.

Analysis of Heavy Metals and Gene Expression of African Arowana Fish (Heterotis niloticus) obtained from Igbalegbe River, Ughelli, Delta State, Nigeria

Heavy metal pollution in aquatic ecosystems can be assessed with suitable biomarkers in fish. This study was carried out to determine the concentration of heavy metals and gene expression of African Arowana Fish (Heterotis niloticus) obtained from Igbalegbe River, Ughelli, Delta State, Nigeria using Atomic Absorption Spectrophotometer (Varian 220 Fast sequential) for the metals and Polymerase chain reaction (PCR) for the gene expression study. Results of the study showed that fishes in the downstream sectionof the river and the effluent discharge point recorded higher metal pollution Index (MPI) compared to the upstream station. The concentration of heavy metals in the fish tissue were generally within the limits of Food and Agricultural Organisation (FAO)/World health Organisation (WHO) except for that of Cadmium and Lead in the discharge station. The selected genes investigated were biomarkers for general stress (HSP70), xenobiotic metabolism (CYPA1), antioxidative defence (SOD), Insulin growth factor-1 (IGF-1) and hypoxia inducible factor (HIF) respectively. The expression of the genes revealed significant variations (P&lt;0.05) in the fishes obtained from the different stations of the river. Some of the genes were down-regulated and suppressed while others were upregulated to enable the fish cope with stress while adapting to environmental pollution. Changes in biomarkers can therefore be considered as early signals of stress in the selected fish species.

Chilled Pacu (Piaractus mesopotamicus) fillets: Modeling Pseudomonas spp. and psychrotrophic bacteria growth and monitoring spoilage indicators by 1H NMR and GC–MS during storage

This study aimed to assess the growth of Pseudomonas spp. and psychrotrophic bacteria in chilled Pacu (Piaractus mesopotamicus), a native South American fish, stored under chilling conditions (0 to 10 °C) through the use of predictive models under isothermal and non-isothermal conditions. Growth kinetic parameters, maximum growth rate (μmax, 1/h), lag time (tLag, h), and (Nmax, Log10 CFU/g) were estimated using the Baranyi and Roberts microbial growth model. Both kinetic parameters, growth rate and lag time, were significantly influenced by temperature (P < 0.05). The square root secondary model was used to describe the bacteria growth as a function of temperature. Secondary models, √μ = 0.016 (T + 10.13) and √μ =0.017 (T + 9.91) presented a linear correlation with R2 values >0.97 and were further validated under non-isothermal conditions. The model's performance was considered acceptable to predict the growth of Pseudomonas spp. and psychrotrophic bacteria in refrigerated Pacu fillets with bias and accuracy factors between 1.24 and 1.49 (fail-safe) and 1.45–1.49, respectively. Fish biomarkers and spoilage indicators were assessed during storage at 0, 4, and 10 °C. Volatile organic compounds, VOCs (1-hexanol, nonanal, octenol, and indicators 2-ethyl-1-hexanol) showed different behavior with storage time (P > 0.05). 1H NMR analysis confirmed increased enzymatic and microbial activity in Pacu fillets stored at 10 °C compared to 0 °C. The developed and validated models obtained in this study can be used as a tool for decision-making on the shelf-life and quality of refrigerated Pacu fillets stored under dynamic conditions from 0 to 10 °C.

Serum Metabolomic Markers of Protein-Rich Foods and Incident CKD: Results From the Atherosclerosis Risk in Communities Study

Rationale & ObjectiveWhile urine excretion of nitrogen estimates total protein intake, biomarkers of specific dietary protein sources have been sparsely studied. Using untargeted metabolomics, this study aimed to identify serum metabolomic markers of six protein-rich foods and to examine whether dietary protein-related metabolites are associated with incident chronic kidney disease (CKD). Study DesignProspective cohort study. Setting & Participants3,726 participants from the Atherosclerosis Risk in Communities (ARIC) study without CKD at baseline. ExposureDietary intake of six protein-rich foods (fish, nuts, legumes, red and processed meat, eggs, poultry), serum metabolites. OutcomesIncident CKD [eGFR <60 mL/min/1.73 m2 with ≥25% eGFR decline relative to visit 1, hospitalization or death related to CKD, or end-stage kidney disease. Analytical ApproachMultivariable linear regression models estimated cross-sectional associations between protein-rich foods and serum metabolites. C-statistics assessed the metabolites’ ability to improve discrimination of highest versus lower three quartiles of intake of protein-rich foods beyond covariates (demographics, clinical factors, health behaviors, and intake of nonprotein food groups). Cox regression models identified prospective associations between protein-related metabolites and incident chronic kidney disease (CKD). ResultsThirty significant associations were identified between protein-rich foods and serum metabolites (fish, n=8; nuts, n=5; legumes, n=0; red and processed meat, n=5; eggs, n=3; poultry, n=9). Metabolites collectively significantly improved discrimination of high intake of protein-rich foods compared to covariates alone (difference in C-statistics=0.033, 0.051, 0.003, 0.024, and 0.025 for fish, nuts, red and processed meat, eggs, and poultry-related metabolites, respectively; p<1.00 x 10-16 for all). Dietary intake of fish was positively associated with 1-docosahexaenoylglycerophosphocholine (22:6n3), which was inversely associated with incident CKD (HR 0.82, 95% CI 0.75-0.89, p=7.81×10-6). LimitationsResidual confounding and sample storage duration. ConclusionsWe identified candidate biomarkers of fish, nuts, red and processed meat, eggs, and poultry. A fish-related metabolite, 1-docosahexaenoylglycerophosphocholine (22:6n3), was associated with lower risk of CKD.

Enzymatic Modifications, as Biomarker for Heavy Metal Toxicity in Catla catla a Fresh Water Fish

Over the last couple of decades, pollution of the environment has grown to be a significant problem, attracting the curiosity of countless experts from developed as well as underdeveloped countries. Sewage dumping into waterways has a detrimental effect on the aquatic ecosystem and biota since it is the principal sink. As a result of their ability to bioaccumulate and intensify as they ascend food chains, toxic metals are not completely eliminated from the environment. For instance, heavy metals have a tendency to accumulate in the muscular tissues of aquatic species; hence, tissues with elevated levels of toxic metals may be detrimental to the well-being of humans and other animals. Fish have been used as indicator organisms to track contamination in aquatic environments. The fish known as catla catla, an Indian Major carp, is edible and extremely susceptible to even the slightest amount of stress. Fish biomarkers are now often employed to assess the internal and exterior health conditions brought on by toxins.&#x0D; The purpose of our investigation was to clarify how biomarkers may be used to track and assess the levels of toxic heavy metals in freshwater Catla catla fish.&#x0D; Results: SOD can be used as a bioindicator for heavy metal pollution, according to the current study's findings. These investigations ultimately aim to quantify pollution concentrations that can cause aquatic environments to undergo permanent biological alterations. Though still mild, the amount of toxicity was approaching to danger. To preserve a healthy and pristine environment, public and government-sponsored initiatives can be undertaken to restrict those activities that discharge contaminants into the ecosystem in an unscientific way.&#x0D; Conclusion: The study comes to the conclusion that in order to evaluate and track the biological state of the aquatic environment, a multiparameter assessment is required.

The use of integrative tools and multiple models for aquatic environmental quality assessment: a case study of the Mirim Lagoon, Southern Brazil.

This study performed toxicity assays with microalgae, microcrustaceans, and fish as well as evaluated biochemical and behavioral biomarkers in fish and microcrustaceans to assess the quality of the surface water of Mirim Lagoon, which belongs to one of the largest hydrographic basins in the world, located in southern Brazil. Three distinct sampling periods were chosen (January, March, and June 2022) based on the rice plantation dynamics which is the main activity surrounding the lagoon. In January, the plantation is irrigated; in March, the water is drained into the Mirim Lagoon, and July is the off-season. Concerning toxicity tests, there was significant inhibition in microalgae growth when exposed to water collected in March, but no mortality was observed for Ceriodaphia dubia, Daphnia magna, and Danio rerio. Regarding biomarkers, behavioral variables contributed more to the higher values of the Integrated Biomarker Response (IBR) index for both D. magna and D. rerio, in March. The Redundancy Analysis (RDA) indicated a correlation between the biomarkers for both organisms and abiotic parameters, mainly nutrients (total phosphorus and total nitrogen), thermotolerant coliforms, total solids, and turbidity. Spatially, there was no difference during monitoring, but the most significant ecotoxicological effects were observed in March. Multivariate analysis and the IBR index proved to be useful tools for monitoring of water bodies such as Mirim Lagoon.

Ecotoxicological Assessment of Ponds Contaminated by Wastewater of Livestock Enterprises by Fish Biomarkers

The morphometric characteristics of internal organs, the morphological composition of blood, and indicators of metabolic processes in the blood plasma and hepatopancreas of the two-year-old carp (&lt;i&gt;Cyprinus carpio&lt;/i&gt; L.) from the conditionally clean pond and contaminated with sewage from a livestock enterprise pond were examined. It was established that the considered contamination caused increase of the spleen mass and its index respectively 1.63 and 1.65 times, but did not affect live weight, body length and height, body structure index, hepatopancreas mass and its index. The hemoglobin concentration, number of erythrocytes, thrombocytes and monocytes in blood of carps from the contaminated pond was lower than in control, whereas number of the segmented neutrophils and ESR were higher, and the number of leukocytes, eosinophils, lymphocytes and rod-nucleated neutrophils did not differ. Wastewater from the livestock enterprise affected metabolic processes in carp, particularly caused increase of the glucose and total protein concentration, the lactate dehydrogenase, aspartate aminotransferase and creatine kinase activity in blood plasma and hepatopancreas. At the same time, there were no differences between specimens from the conditionally clean and contaminated pond in terms of content of triglycerols, cholesterol, urea, creatinine, calcium, inorganic phosphorus, AlAT and AsAt activity in blood plasma and hepatopancreas and total protein content and LHD activity in hepatopancreas, content of total protein, glucose, creatinine, urea, the glucose content in hepatopancreas. The established changes in internal organs, morphological composition of blood and metabolic parameters, particularly concentration of hemoglobin and the erythrocytes number in blood, glucose and total protein content, and activity of lactate dehydrogenase and creatine kinase in carp enabled to propose to use these parameters as supplementary over assessment of the ecotoxicological state of ponds contaminated by wastewater from livestock enterprises.

Accelerated eutrophication alters fish and aquatic health: a quantitative assessment by using integrative multimarker, hydrochemical, and GIS modelling method in an urban lake.

The ramifications of anthropogenic activities on the environment and the welfare of aquatic life in lakes worldwide are becoming increasingly alarming. There is a lack of research in the Indian Himalayas on fish biomarker responses to stressful aquatic conditions and the use of environmetric modelling in GIS. Our research evaluates the environmental health of urban lakes in multiple basins using multi-biomarker endpoints (13 features) in Schizothorax niger and hydrochemical characterization (9 features) of water. The study covers 31 grids, each at a distance of 1 km2. This study demonstrated a statistically significant (P = 0.001) increase in white blood cells (WBC), mean cell size (MCH), helminth infection, and health assessment index score (HAIS) score in fish from a highly eutrophic cluster or basin compared to a reference cluster, which is indicative of environmental stress in fish. Based on hydrochemical similarities, the lake water datasets were divided into three categories using hierarchical cluster analysis (HCA). In the PCA analysis, the first three principal components were responsible for 78.1% of the data's variance. The first principal component (PC1) accounted for 57.4% of the variance and had a strong positive loading from ammonia, total phosphate, pH, nitrates, and total alkalinity for water quality parameters. Additionally, PC1 had a favourable loading from WBC, helminth infection (%), and the health assessment index score (HAIS) for biological endpoints. These findings are in alignment with the results of the multivariate analysis. The trophic state index (TSI) showed a significant (P < 0.05) increase in Cluster 1, which includes the peripheral areas of Hazratbal and Gagribal side (> 70), compared to the reference cluster. The multiple regression model indicates that ammonia, phosphate, and nitrate significantly impact the general health of fish (R2 > 0.7). A novel methodology for monitoring water quality fluctuations across different basins and clusters is presented in this study. By integrating fish health biomarkers and GIS technology, we have developed a comprehensive approach to evaluate the overall well-being of aquatic habitat. This technique may prove beneficial in the management of urban lentic water bodies in the Kashmir Himalayas and other comparable water systems around the globe, while also supporting sustainable practices.

Specific Blood Plasma Circulating miRs Are Associated with the Physiological Impact of Total Fish Meal Replacement with Soybean Meal in Diets for Rainbow Trout (Oncorhynchus mykiss).

High dietary SBM content is known to induce important physiological alterations, hampering its use as a major FM alternative. Rainbow trout (Oncorhynchus mykiss) juveniles were fed two experimental diets during 9 weeks: (i) a FM diet containing 12% FM; and (ii) a vegetable meal (VM) diet totally devoid of FM and based on SBM (26%). Fish fed the VM diet did not show reduced growth performance when compared with fish fed the FM diet. Nevertheless, fish fed the VM diet had an increased viscerosomatic index, lower apparent fat digestibility, higher aminopeptidase enzyme activity and number of villi fusions, and lower α-amylase enzyme activity and brush border integrity. Small RNA-Seq analysis identified six miRs (omy-miR-730a-5p, omy-miR-135c-5p, omy-miR-93a-3p, omy-miR-152-5p, omy-miR-133a-5p, and omy-miR-196a-3p) with higher expression in blood plasma from fish fed the VM diet. Bioinformatic prediction of target mRNAs identified several overrepresented biological processes known to be associated with high dietary SBM content (e.g., lipid metabolism, epithelial integrity disruption, and bile acid status). The present research work increases our understanding of how SBM dietary content has a physiological impact in farmed fish and suggests circulating miRs might be suitable, integrative, and less invasive biomarkers in fish.

Sex-biased response of pollution biomarkers in fish: Insights from the killifish Poecilia vivipara

Copper ions (Cu) are one of the most frequent trace-contaminants found in Brazilian waters and, although considered as an essential element, in high concentrations can accumulate and induce toxicity. Biomarkers are important tools that can be used to assess these impacts, but to be considered trustworthy, they have to be previously tested in target organisms through laboratory studies under controlled conditions. However, many of these experiments are conducted using only males, as it is believed that the hormonal variation of females can bias the results, increasing data variability. Notwithstanding, few studies have actually tested this hypothesis, highlighting the importance of considering and measuring the role of sex in ecotoxicological studies. The aim this study was to evaluate the influence of sex on biomarkers classically used in environmental monitoring programs using the fish Poecilia vivipara as model. For this, females and males were exposed for 96 h to two Cu concentrations (9 and 20 μg/L) and a control group. In liver and gills, Cu accumulation, total antioxidant capacity (TAC) and lipid peroxidation (LPO) were evaluated. In addition, samples of peripheral blood were used for neutrophil to lymphocyte ratio determination, a measure of the onset of secondary stress. Results show that Cu hepatic accumulation did not differ between females and males, but higher levels of this metal were observed in exposed animals compared to control fish. Additionally, interactive effects were observed for hepatic LPO, as males showed elevated oxidative damage in comparison to females. Moreover, Cu exposure elevated hepatic LPO relative to control only in males, but this increase in oxidative damage was not accompanied by changes in liver TAC. On the other hand, differences in branchial Cu accumulation and LPO were not observed. Conversely, control females showed elevated TAC in comparison to control males, but Cu exposure eliminated this difference. Cu exposure also induced an increase in the N:L ratio, indicating the presence of a secondary stress response unrelated to sex. Ultimately, the findings of this study demonstrate that sex can influence the response of biomarkers that are typically used in ecotoxicological investigations in a multifaceted manner. As a result, using animals from a singular sex in such studies may result in consequential outcomes, potentially leading to underestimation or overestimation of results.

Acute exposure to realistic concentrations of Bisphenol-A trigger health damage in fish: Blood parameters, gene expression, oxidative stress

Despite much information regarding BPA toxicity in fish and other aquatic organisms, data is still misleading as most studies have utilized concentrations several orders of magnitude higher than those typically found in the environment. As an illustration, eight of the ten studies investigating the impact of BPA on the biochemical and hematological parameters of fish have employed concentrations on the order of mg/L. Therefore, the results may not accurately represent the effects observed in the natural environment. Considering the information above, our study aimed to 1) determine whether or not realistic concentrations of BPA might alter the biochemical and blood parameters of Danio rerio and trigger an inflammatory response in the fish liver, brain, gills, and gut and 2) determine which organ could be more affected after exposure to this chemical. Findings pinpoint that realistic concentrations of BPA prompted a substantial increase in antioxidant and oxidant biomarkers in fish, triggering an oxidative stress response in all organs. Likewise, the expression of different genes related to inflammation and apoptosis response was significantly augmented in all organs. Our Pearson correlation shows gene expression was closely associated with the oxidative stress response. Regarding blood parameters, acute exposure to BPA generated biochemical and hematological parameters increased concentration-dependent. Thus, it can be concluded that BPA, at environmentally relevant concentrations, threatens aquatic species, as it prompts polychromasia and liver dysfunction in fish after acute exposure.

Revealing the environmental pollution of two estuaries through histopathological biomarkers in five fishes from different trophic guilds of northeastern Brazil

Estuaries in Brazil are mostly anthropically affected due to the discharge of industrial and domestic effluents. In two of them, the Santa Cruz Channel Estuary (ITAP) and Sirinhaém River Estuary (SIR), historically affected by mercury pollution and sugarcane industry in Northeast Brazil, we assessed environmental pollution using liver and gill histopathological biomarkers in fish from different trophic levels. Liver samples exhibited serious damages such as hepatic steatosis, necrosis, and infiltration. The gills showed moderate to severe changes, such as lifting of epithelial cells, lamellar aneurysm, and rupture of lamellar epithelium. Most of the changes in the liver and gills were reported for species Centropomus undecimalis and the Gobionellus stomatus, which were considered as good sentinels of pollution. The combination of biomarker methodologies was efficient in diagnosing the serious damage to the species, reinforcing the need for monitoring the health of the ecosystems evaluated.