Collective movement of schooling fish reduces the costs of locomotion in turbulent conditions.

The ecological and evolutionary benefits of energy-saving in collective behaviors are rooted in the physical principles and physiological mechanisms underpinning animal locomotion. We propose a turbulence sheltering hypothesis that collective movements of fish schools in turbulent flow can reduce the total energetic cost of locomotion by shielding individuals from the perturbation of chaotic turbulent eddies. We test this hypothesis by quantifying energetics and kinematics in schools of giant danio (Devario aequipinnatus) and compared that to solitary individuals swimming under laminar and turbulent conditions over a wide speed range. We discovered that, when swimming at high speeds and high turbulence levels, fish schools reduced their total energy expenditure (TEE, both aerobic and anaerobic energy) by 63% to 79% compared to solitary fish (e.g., 228 versus 48 kj kg-1). Solitary individuals spend approximately 22% more kinematic effort (tail beat amplitude•frequency: 1.7 versus 1.4 BL s-1) to swim in turbulence at higher speeds than in laminar conditions. Fish schools swimming in turbulence reduced their three-dimensional group volume by 41% to 68% (at higher speeds, approximately 103 versus 33 cm3) and did not alter their kinematic effort compared to laminar conditions. This substantial energy saving highlights that schooling behaviors can mitigate turbulent disturbances by sheltering fish (within schools) from the eddies of sufficient kinetic energy that can disrupt locomotor gaits. Therefore, providing a more desirable internal hydrodynamic environment could be one of the ecological drivers underlying collective behaviors in a dense fluid environment.

Assessment of trace and macroelement accumulation in cyprinid juveniles as bioindicators of aquatic pollution: effects of diets and habitat preferences.

Juveniles of three cyprinids with various diets and habitat preferences were collected from the Szamos River (Hungary) during a period of pollution in November 2013: the herbivorous, benthic nase (Chondrostoma nasus), the benthivorous, benthic barbel (Barbus barbus), and the omnivorous, pelagic chub (Squalius cephalus). Our study aimed to assess the accumulation of these elements across species with varying diets and habitat preferences, as well as their potential role in biomonitoring efforts. The Ca, K, Mg, Na, Cd, Cr, Cu, Fe, Mn, Pb, Sr, and Zn concentration was analyzed in muscle, gills, and liver using MP-AES. The muscle and gill concentrations of Cr, Cu, Fe, and Zn increased with trophic level. At the same time, several differences were found among the trace element patterns related to habitat preferences. The trace elements, including Cd, Pb, and Zn, which exceeded threshold concentrations in the water, exhibited higher accumulations mainly in the muscle and gills of the pelagic chub. Furthermore, the elevated concentrations of trace elements in sediments (Cr, Cu, Mn) demonstrated higher accumulation in the benthic nase and barbel. Our findings show habitat preference as a key factor in juvenile bioindicator capability, advocating for the simultaneous use of pelagic and benthic juveniles to assess water and sediment pollution status.

Bioaccumulation of arsenic in fish (Labeo rohita) in presence of periphyton: ameliorative effect on oxidative stress, physiological condition, immune response and risk assessment.

The present study explores the use of periphyton to ameliorate toxic properties of arsenic (As) to Labeo rohita and also assesses the human food safety aspects. Fish were introduced to arsenite [As(III)] contaminated water (0.3 and 3 mg/L) along with periphyton. Biochemical, physiological and immunological parameters, including gene expression, were assessed after 30 days of exposure. Periphyton incorporation significantly improved (p < 0.05) the adverse effects of As on respiration, NH3 excretion and brain AChE activity by reducing oxidative stress and As bioaccumulation. The presence of periphyton in As(III) exposed fish (3 mg/L) increased the immune response (Immunoglobulin M and Complement C3) in the serum and the regulation of the respective immune genes in the anterior kidney was found to be similar to the control. A speciation study using LC-ICP-MS confirmed the high accumulation of As by periphyton (5.0-31.9 µg/g) as arsenate [As (V)], resulting in a lower amount of As in fish muscle. The calculated human health risk indices, Target Hazard Quotient (THQ) and Target Cancer risk (TCR) indicate that fish grown in periphyton-treated water may lower the human health risks associated with As. The study signifies the importance of periphyton-based aquaculture systems in As contaminated regions for safe fish production with enhanced yield.

Antibiotic residues in commercial freshwater fish from southeast China: distribution and human health risk assessment.

We investigated 14 antibiotic residues in 8 marketed freshwater fish species from southeast China and estimated the associated health risks to local consumers. The antibiotic residues were determined by UPLC-MS/MS. Our findings revealed widespread distribution of quinolones (QNs), tetracyclines (TCs), and chloramphenicols (CAPs) in the freshwater fish. Notably, the average concentrations of enrofloxacin and ciprofloxacin reached levels as high as 62.5 µg/kg wet weight (ww) and 11.7 µg/kg ww, respectively, and detection frequencies were 68.7% for enrofloxacin and 31.6% for ciprofloxacin. Additionally, we detected chloramphenicol, a prohibited antibiotic, in samples with a detection frequency of 0.76%. Among the fish species, the mean concentration of total antibiotic residues was highest in bluntnose black bream (263.3 µg/kg), followed by English perch (52.4 µg/kg), crucian carp (46.3 µg/kg), black carp (28.6 µg/kg), yellowcheek carp (21.0 µg/kg), grass carp (15.3 µg/kg), bighead carp (3.78 µg/kg), and mandarin fish (3.69 µg/kg). We estimated the daily intake values of these antibiotic residues which were lower than the acceptable daily intake values and hazard indexes were much less than 1. It indicates that there is very low direct health risk to consumers. Despite that, investigation on the chronic impact, such as antibiotic-resistant bacteria, gut microbiota disruption, and allergic reactions, is urgently needed.

Assessment of microplastic contamination in some commercial fishes of the southern Caspian Sea and its potential risks.

This study examined the occurrence of microplastics (MPs) in the gastrointestinal tract (GIT) of 384 fishes classified into four species from 11 sites in 2022 from the southern part of the Caspian Sea. GITs of fishes were collected and digested in H2O2 and KOH at 45 °C for 72 h. After filtration, extracted MPs were observed under a stereomicroscope, and selected MP particles were identified using FTIR. Presence of MPs was 68.98% in the GIT of the investigated fish. The mean abundance of MPs was 5.9 ± 0.9 MPs/GIT in Rutilus kutum, 9.2 ± 1.2 MPs/GIT in Chelon auratus, 3.6 ± 0.7 MPs/GIT in Alosa braschnikowi, and 2.7 ± 0.5 MPs/GIT in Vimba vimba. The predominant form of MPs was fiber (58.21%), followed by fragment (34.77%). Black (34.4%), white (19.07%), and blue (14.58%) were the most frequently detected colors of MPs. Overall, 6 MP polymers were identified, dominantly polypropylene (42.86%), polystyrene (17.86%), and cellophane (14.28%). The western part of the Caspian Sea (mostly tourist spots and urban areas) showed more MP pollution in fish compared to the eastern part. Polymer hazard index (PHI) revealed alarming microplastic contamination in the southern Caspian Sea. The PHI value of the present study showed that PES (PHI = 8403.78) and PS (PHI = 535.80) were "Extreme danger" and "Danger" risk categories, respectively.

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.

Integrating fish swimming abilities into rapid road crossing barrier assessment: Case studies in the southeastern United States.

Many aquatic networks are fragmented by road crossing structures; remediating these barriers to allow fish passage is critical to restoring connectivity. Maximizing connectivity requires effective barrier identification and prioritization, but many barrier prioritization efforts do not consider swimming capabilities of target species. Given the many potential barriers within watersheds, inventory efforts integrating species-specific swimming speeds into rapid assessment protocols may allow for more accurate barrier removal prioritization. In this study, we demonstrate an approach for integrating fish swimming speeds into rapid barrier assessment and illustrate its utility via two case studies. We measured critical swimming speeds (Ucrit) of two stream-resident fish species with very different swimming modes: Yoknapatawpha Darter (Etheostoma faulkneri), an at-risk species whose current distribution is restricted to highly degraded habitat, and Bluehead Chub (Nocomis leptocephalus), an important host species for the federally endangered Carolina Heelsplitter mussel (Lasmigona decorata). We assessed potential barriers for Yoknapatawpha Darters in the Mississippi-Yocona River watershed, and Bluehead Chubs in the Stevens Creek watershed, South Carolina, USA. We integrated Ucrit into the Southeast Aquatic Resources Partnership (SARP) barrier assessment protocol by estimating the proportion of individuals per species swimming at least as fast as the current through the assessed structures. Integrating Ucrit estimates into the SARP protocol considerably increased barrier severity estimates and rankings only for Yoknapatawpha Darters in the Yocona River watershed. These results indicate the importance of including species-specific swimming abilities in rapid barrier assessments and the importance of species-watershed contexts in estimating where swimming speed information might be most important. Our method has broad application for those working to identify barriers more realistically to improve species-specific fish passage. This work represents a next step in improving rapid barrier assessments and could be improved by investigating how results change with different measurements of swimming abilities and structure characteristics.

Trace elements in the muscle and liver tissues of Garra shamal from the freshwater ecosystem of Oman: an exposure risk assessment.

Anthropogenic activities lead to environmental contamination with foreign substances such as heavy metals. This work was aimed to monitor trace elements (total arsenic (As), cadmium (Cd), chrome (Cr), cobalt (Co), copper (Cu), lead (Pb), manganese (Mn), mercury (Hg), nickel (Ni), and zinc (Zn)) contamination levels (dry weight base) in three natural freshwater reservoirs of Oman including Al Khawd and Al Amarat (Muscat Governorate) and Surur area (Ad Dakhiliyah Governorate as control area) using a native benthic inland fish (Garra shamal; Cyprinidae) for the first time. The muscle and liver of a hundred and twenty G. shamal were collected to assess the degree of metal contamination. Atomic absorption spectrometry was used as an analytical technique. From the spectrum of analyzed elements, we found Zn as a major element in monitored areas. The statistically significant (P < 0.05) highest concentrations of Zn liver (0.275 ± 0.065 µg/g) were in Al Amarat compared to the other areas. The concentrations of monitored elements in the fish muscle were lower than the liver samples. Furthermore, the fish length was significantly correlated with the accumulation of Hg and Co in both muscle and liver samples. In all analyzed fish from Oman inland water, the concentrations of elements were below the permissible limits; however, additional research is needed.

Assessment of imidacloprid induced genotoxicity in Pethia conchonius (Rosy barb), a common freshwater fish of India.

Imidacloprid is one of the highly efficient, globally used neonicotinoid groups of insecticides. The indiscriminate use of imidacloprid is contaminating large water bodies affecting not only the target organisms but also non-target organisms including fish. The present study aimed to assess the extent of nuclear DNA damage by imidacloprid in Pethia conchonius a freshwater fish in India using comet and micronucleus assays. The LC50 value of imidacloprid was estimated to be 227.33 mg L-1. Based on the LC50-96 h value, three sub-lethal concentrations of imidacloprid, SLC I -18.94 mg L-1, SLC II -28.41 mg L-1 and SLC III -56.83 mg L-1 were used to detect its genotoxic effect at DNA and cellular level. The imidacloprid exposed fishes exhibited higher DNA damage and nuclear abnormalities (p < 0.05) than the control. The %head DNA, %tail DNA, tail length and the frequency of micronuclei with other nuclear abnormalities like blebbed and notched nuclei were significantly higher than the control in a time and concentration-dependent manner. The DNA damage parameters such as %head DNA (29.107 ± 1.843), %tail DNA (70.893 ± 1.843), tail length (361.431 ± 8.455) micronucleus (1.300 ± 0.019), notched (0.844 ± 0.011) and blebbed (0.811 ± 0.011) nuclei were found to be highest for SLC III (56.83 mg L-1) at 96 h. The findings indicate that IMI is highly genotoxic in fish and other vertebrates leading to mutagenic/clastogenic effects. The study will be helpful in optimization of the imidacloprid use.

Assessment and Bioaccumulation of Heavy Metals in Water, Fish (wild and Farmed) and Associated Human Health Risk.

Worldwide, anthropogenic activities are causing the natural environment and aquaculture systems to become heavily contaminated with heavy metals, which can lead to consumer's health problems. In the current study, wild and farmed fish (n = 30) and water samples (n = 6) have been collected from the Chashma barrage and fish farm to assess the heavy metals concentration, i.e., Cu, Cd, Pb, Zn and Cr, in the water and some important organs (gills, liver, muscle, brain and bones) of wild and farmed fish (Labeo rohita) using Graphite furnace Atomic absorption spectrometry. Bioaccumulation factor and human health risk assessment were calculated to measure the health status of both fish and humans. Results show that in wild and farm fish's gills, muscles and bones, the trend of the heavy metals was Zn > Pb > Cu > Cd > Cr. On the other hand, the brain and liver show Zn > Cu > Pb > Cd > Cr trend. Comparatively, the heavy metals concentration was mostly higher (P < 0.05) in wild fish. Further; in both fish habitats (water) the heavy metals (Cd and Pb) concentration was higher than the WHO standard level, while in the body, Cd was higher (P < 0.05) in all studied organs except the muscle, Cr was only lower (P > 0.05) in muscle and brain. Pb was higher (P < 0.05) in all studied organs of both fish. Bioaccumulation of heavy metals was mostly higher (P < 0.05) in wild fish than in farmed. EDI and THQ were higher in wild fish, but the HI value was lower than 1 for both fish. Moreover, the PCA analysis suggests a positive correlation between heavy metals concentration in fish organs and the water of both fish (wild and farmed). Results indicated that farmed fish showed less potential risk to humans than wild fish.

Heavy metal concentrations in various tissues of two fish species from Damsa Dam Lake (Turkey) and associated health risk assessment.

PURPOSE OF REVIEW AND METHOD: In this study, indicate the average or range of 10 heavy metal concentrations in muscle, gills and liver of two fish species (Squalius pursakensis and Cyprinus carpio) living in the Damsa Dam Lake (Turkey) was examined in relation to fish gender and size. Inductively coupled plasma-tandem mass spectrometry was used to evaluate the concentrations of heavy metals. The average contents of the metals in S. pursakensis muscle samples were 0.877, 9.285, 0.614, 0.488, 8.190, 0.155, 0.015, 0.010, 0.218, 0.009 mg/kg for Cu, Fe, Mn, Ni, Zn, As, Cd, Co, Cr and Pb respectively. On the other hand, the average contents of the metals in C. carpio muscle samples were 0.673, 8.001, 0.367, 0.435, 6.107, 0.188, 0.007, 0.007, 0.371, 0.012 mg/kg for Cu, Fe, Mn, Ni, Zn, As, Cd, Co, Cr and Pb respectively. In addition, human health risks were evaluated considering the estimated daily intake of heavy metals in adults. The target hazard quotient, hazard index and carcinogenic risk values were determined based on the frequency of fish consumption for the population living in the region. RESULT AND CONCLUSION: The highest quantities of all heavy metals were determined in the gills and liver of the fish species. The levels of most heavy metals in tissues did not differ significantly between both fish species. Only a few heavy metals in the tissues of both fish species demonstrated significant associations with fish length or weight. In terms of the levels of most of the heavy metals in the tissues, there weren't any significant differences between the male and the female fish. The estimated daily intakes of all of the heavy metals were lower than the reference doses. The values of the target hazard quotient and the hazard index were found to be below 1 and the values regarding carcinogenic risk were assessed to be within or lower than the permissible levels. Cr, Cu, Zn, Cd, Pb and inorganic As values assessed in the muscle tissues of fish were found below the values acceptable by national and international institutions. These results proved that the examined fish species can be consumed safely.

Hazard assessment of complex legacy-contaminated groundwater mixtures using a novel approach method in adult fathead minnows.

Traditional risk assessment methods face challenges in the determination of drivers of toxicity for complex mixtures such as those present at legacy-contaminated sites. Bioassay-driven analysis across several levels of biological organization represents an approach to address these obstacles. This study aimed to apply a novel transcriptomics tool, the EcoToxChip, to characterize the effects of complex mixtures of contaminants in adult fathead minnows (FHMs) and to compare molecular response patterns to higher-level biological responses. Adult FHMs were exposed for 4 and 21 days to groundwater mixtures collected from a legacy-contaminated site. Adult FHM showed significant induction of micronuclei in erythrocytes, decrease in reproductive capacities, and some abnormal appearance of liver histology. Parallel EcoToxChip analyses showed a high proportion of upregulated genes and a few downregulated genes characteristic of compensatory responses. The three most enriched pathways included thyroid endocrine processes, transcription and translation cellular processes, and xenobiotics and reactive oxygen species metabolism. Several of the most differentially regulated genes involved in these biological pathways could be linked to the apical outcomes observed in FHMs. We concluded that molecular responses as determined by EcoToxChip analysis show promise for informing of apical outcomes and could support risk assessments of complex contaminated sites.

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.

Fishery, growth, mortality, and stock assessment of endangered Tor putitora from Tehri dam reservoir, Uttarakhand, Himalayan foothills of India in relation to environmental variables.

The current study determined Tor putitora (Hamilton, 1822) fishery, growth, mortality, and population characteristics using length-frequency data assembled monthly from the Tehri dam reservoir in Uttarakhand from January to December 2022. The estimation data was separated into 40-mm class intervals, and population parameters were investigated and computed using the FiSAT-II software tool. W = 0.0101 L2.996, where a = 0.0101 and b = 2.99, were determined as the length-weight relationships, and the growth performance index (ϕ) was computed to be 5.40. Tor putitora commercial catches in the Tehri dam reservoir were dominated by length groups of 360-399 and 320-359 mm. Different growth parameters were estimated using length-frequency data as L∞ = 987.00 mm, K = 0.26 yr-1, and t0 = -0.0003 years. Z, M, and F mortality coefficients were estimated to be 1.01, 0.27, and 0.73, respectively. At the end of the first, second, third, fourth, fifth, sixth, and seventh years, the fish measured 226, 400, 535, 638, 718, and 780, and 827, respectively. The estimated value of the exploitation rate (E) was 0.73 using the length-converted catch curve approach, which was determined to be somewhat higher than the optimum value (0.50). Tor putitora recruitment patterns from the Tehri dam reservoir reveal that the species only has one recruitment pattern every year, and that solely occurs from June to September. The current exploitation level (0.73) has already exceeded the maximum fishing pressure (Emax = 0.508), indicating that there is a decline in the catch at the current fishing pressure, and a further increase in fishing efforts may lead to a decline in the stock, which may be detrimental to the sustainable fishery of Tor putitora in the Tehri dam reservoir, Uttarakhand, India.

Expanding non-invasive approaches for fish-health monitoring: A survey of the epidermal mucous metabolomes of phylogenetically diverse freshwater fish species.

There is a pressing need for more-holistic approaches to fisheries assessments along with growing demand to reduce the health impacts of sample collections. Metabolomic tools enable the use of sample matrices that can be collected with minimal impact on the organism (e.g., blood, urine, and mucus) and provide high-throughput, untargeted biochemical information without the requirement of a sequenced genome. These qualities make metabolomics ideal for monitoring a wide range of fish species, particularly those under protected status. In the current study, we surveyed the relative abundances of 120 endogenous metabolites in epidermal mucus across eight freshwater fish species belonging to seven phylogenetic orders. Principal component analysis was used to provide an overview of the data set, revealing strong interspecies relationships in the epidermal mucous metabolome. Normalized relative abundances of individual endogenous metabolites were then used to identify commonalities across multiple species, as well as those metabolites that showed notable species specificity. For example, taurine was measured in high relative abundance in the epidermal mucus of common carp (Cyprinus carpio), northern pike (Esox lucius), golden shiner (Notemigonus crysoleucas), rainbow trout (Oncorhynchus mykiss), and rainbow smelt (Osmerus mordax), whereas γ-amino butyric acid (GABA) exhibited a uniquely high relative abundance in flathead catfish (Pylodictis olivaris). Finally, hierarchical cluster analysis was used to evaluate species relatedness as characterized by both the epidermal mucous metabolome (phenotype) and genetic phylogeny (genotype). This comparison revealed species for which relatedness in the epidermal mucous metabolome composition closely aligns with phylogenetic relatedness (e.g., N. crysoleucas and C. carpio), as well as species for which these two measures are not well aligned (e.g., P. olivaris and Polyodon spathula). These, and other findings reported here, highlight novel areas for future research with fish, including development of epidermal mucous-based markers for non-invasive health monitoring, sex determination, and hypoxia tolerance.

Development and Initial Testing of EcoToxChip, a Novel Toxicogenomics Tool for Environmental Management and Chemical Risk Assessment.

New approach methods (NAMs) are increasingly important to help accelerate the pace of ecological risk assessment and offer more ethical, affordable, and efficient alternatives to traditional toxicity tests. In the present study, we describe the development, technical characterization, and initial testing of a toxicogenomics tool, EcoToxChip (384-well quantitative polymerase chain reaction [qPCR] array), to support chemical management and environmental monitoring for three laboratory model species-fathead minnow (Pimephales promelas), African clawed frog (Xenopus laevis), and Japanese quail (Coturnix japonica). Chip design, including gene selection, was informed by a diverse end-user group and quality control metrics (e.g., primer assay, reverse transcription, and PCR efficiency) performed well based on a priori established criteria. Correlation with RNA sequencing (seq) data provided additional confidence in this novel toxicogenomics tool. Although the present study represents an initial testing of only 24 EcoToxChips for each of the model species, the results provide increased confidence in the robustness/reproducibility of EcoToxChips for evaluating perturbations in gene expression associated with chemical exposure and thus, this NAM, combined with early-life stage toxicity testing, could augment current efforts for chemical prioritization and environmental management. Environ Toxicol Chem 2023;42:1763-1771. © 2023 SETAC.

Assessment of ichthyofaunal diversity of family Cyprinidae in River Panjkora Dir, Khyber Pakhtunkhwa, Pakistan.

The study explores the Ichthyofaunal diversity of freshwater Cyprinidae fish in the Panjkora River of Dir, Khyber Pakhtunkhwa, Pakistan. A total of 911 fish samples were collected from 5 different sites along the river and identified using standard identification keys. Out of the 911 fish samples, 204 were related to family Cyprinidae, having 4 sub families, i.e Cyprininae representing 3 species i.e Carassius auratus, Cyprinus carpio, Cyprinus watsoni, Danioninae representing 6 species i.e Barilius modestus, Barilius vagra, Barilius pakistanicus, Amblypharangodon mola, Salmophasia punjabensis, Rasbora daniconius, Labeoninae representing 5 species i.e Tor putitora, Crossocheilus diplocheilus, Gara gotyla, Labeo rohita, Labeo diplocheilus, and Barbinae representing 6 species i.e Puntius ticto, Puntius sophore, Puntius chonchonius, Schizothorax esocinus, Schizothorax labiatus and Schizothorax plagiostomus. This study adds 6 new species i.e Amblypharangodon mola, Salmophasia punjabensis, Labeo rohita, Labeo diplocheilus, Puntius chonchonius and Rasbora daniconius for the 1st time which are not reported earlier. EXCEL 2019 and XLSTAT were used for principal components analysis to study richness and correlation of fish diversity. Eigenvalues obtained from Sheringal to Zulam are 3.72, 0.511, 0.37, 0.29 and 0.09 respectively. The higher eigenvalue of Sheringal site shows higher diversity. Water quality study shows that the mean values of water temperature was 16.8°C, pH 7.08, total hardness134.6 mg/L, alkalinity 89.3 mg/L, Nitrate 5.93 mg/L, Chlorine 0.87 mg/L, Nitrite0.02 mg/L and Copper 0.04 mg/L, Iron 0.07 mg/L, Lead 0.002 mg/L. The physicochemical parameters of river Panjkora during the study time remained within the permissible limits as prescribed by the WHO (2005). Gut analysis revealed the presence of algae, nymphs insect's larvae, macroinvertebrates and protozoans.

Mitochondrial DNA reveals shallow population genetic structure in economically important Cyprinid fish Labeo rohita (Hamilton, 1822) from South and Southeast Asia.

BACKGROUND: Assessment of genetic diversity and population genetic structure is important for species that are economically important, threatened, and are at global conservation priority. Analysis of mitochondrial DNA is broadly used in species identification and population genetics studies due to the availability of sufficient reference data and better evolutionary dynamics for phylogeographic investigation. Labeo rohita (Rohu) is an economically important species cultured under carp polyculture systems in Asia. The present study explores the genetic diversity, phylogeography, and population structure of L. rohita from different countries using cytochrome oxidase subunit I (COI) gene. METHODS AND RESULTS: A total of 17 L. rohita specimens were sampled from River Beas, India. For the genetic study, we amplified and sequenced COI mitochondrial DNA region. The obtained genetic data was combined with 268 COI records available in the NCBI and BOLD databases originating from multiple populations/countries across South and Southeast Asia. As a result, 33 haplotypes were identified that displayed low nucleotide (π = 0.0233) and moderate haplotype diversity (Hd = 0.523). Tajima (D) was found to be negative (P > 0.05), whereas Fu's Fs showed a positive value (P > 0.05). The overall FST value between studied populations was 0.481 (P < 0.05). CONCLUSION: AMOVA analysis indicated higher variation within than among the population examined. The neutrality tests suggested the presence of rare haplotypes and stable demography within studied populations of L. rohita. The Bayesian skyline plot indicated steady population growth until 1 Mya followed by population decline, whereas FST values indicated significant genetic differentiation. High heterogeneity was observed in the Pakistan population which could be indicative of long-term isolation and excessive culturing to meet market demands. The present results are the first global comparative analysis of L. rohita and pave the way forward for detailed genomic and ecological studies aimed at the development of improved stock and effective conservation plans. The study also makes recommendations to conserve the genetic integrity of wild species from aquaculture-reared fishes.

Ecotoxicological QSAR study of fused/non-fused polycyclic aromatic hydrocarbons (FNFPAHs): Assessment and priority ranking of the acute toxicity to Pimephales promelas by QSAR and consensus modeling methods.

Fused/non-fused polycyclic aromatic hydrocarbons (FNFPAHs) have a variety of toxic effects on ecosystems and human body, but the acquisition of their toxicity data is greatly limited by the limited resources available. Here, we followed the EU REACH regulation and used Pimephales promelas as a model organism to investigate the quantitative structure-activity relationship (QSAR) between the FNFPAHs and their toxicity for the aquatic environment for the first time. We developed a single QSAR model (SM1) containing five simple and interpretable 2D molecular descriptors, which met the validation of OECD QSAR-related principles, and analyzed their mechanistic relationships with toxicity in detail. The model had good degree of fitting and robustness, and had better external prediction performance (MAEtest = 0.4219) than ECOSAR model (MAEtest = 0.5614). To further enhance its prediction accuracy, the three qualified single models (SMs) were used for constructing consensus models (CMs), the best one CM2 (MAEtest = 0.3954) had a significantly higher prediction accuracy for test compounds than SM1, and also outperformed the T.E.S.T. consensus model (MAEtest = 0.4233). Subsequently, the toxicity of 252 true external FNFPAHs from Pesticide Properties Database (PPDB) was predicted by SM1, the prediction results showed that 94.84 % compounds were reliably predicted within the model's application domain (AD). We also applied the best CM2 to predict the untested 252 FNFPAHs. Furthermore, we provided a mechanistic analysis and explanation for pesticides ranked as top 10 most toxic FNFPAHs. In summary, all developed QSAR and consensus models can be used as efficient tools for predicting the acute toxicity of unknown FNFPAHs to Pimephales promelas, thus being important for the risk assessment and regulation of FNFPAHs contamination in aquatic environment.

Accumulation and human health risk assessment of trace elements in two fish species, Cirrhinus mrigala and Oreochromis niloticus, at Tarukri Drain, District Rahimyar Khan, Punjab, Pakistan.

The toxic discharge of heavy metals into the water affects the aquatic ecosystem as well as the human population interacting with it because of their toxicity, bioaccumulation, long persistence, and transfer through the food chain. Thus, it is very important to conduct studies to determine the level of heavy metal pollution in order to better control, manage, and preserve the pollution of aquatic ecosystems. This study assessed heavy metal contamination in fish and its associated health risk to the population around the Tarukri Drain, Punjab, Pakistan. Two fish species (Oreochromis niloticus and Cirrhinus mrigala) were collected from three different sites in two different seasons. Collected fish were analyzed for cadmium (Cd), iron (Fe), lead (Pb), nickel (Ni), and zinc (Zn) accumulation using atomic absorption spectrometer (AAS). Furthermore, the human health risks associated with the consumption of affected fish were also assessed. Target hazardous quotient for seasonal consumers was between 0.12, - 1.58 × 10-4, and 0.54 - 3.28 × 10-4 (mg/kg) in O. niloticus and C. mrigala, respectively. While for regular consumers it was between 0.28-3.71 × 10-4 and 1.27-7.68 × 10-4 (mg/kg) in O. niloticus and C. mrigala respectively for the studied heavy metals. Fish sampled from Sadiqabad contained the highest concentration of heavy metals. The analysis of fish organs (kidney, liver, and muscles) showed heavy metal accumulation in the order of kidney > liver > muscles (p < 0.00). The obtained results showed that heavy metal contaminations in both fish species were within the permissible limits recommended by the World Health Organization (WHO). Both sampling seasons (i.e., summer and winter) showed a non-significant difference in heavy metal concentration. The calculated total target hazardous quotient across all heavy metals remained < 1 with only one exception. The carcinogenic risk assessment of heavy metals showed a non-significant effect in both fish species.