Application of a new approach method to assess the hazard of complex legacy contaminated groundwater mixtures on fathead minnows in outdoor mesocosms.

Assessing the environmental risks of contaminated groundwater presents significant challenges due to its often-complex chemical composition and to dynamic processes affecting exposure of organisms in receiving surface waters. The objective of this study was to characterize the effects of groundwater collected from a legacy contaminated industrial site, in fish under environmentally relevant conditions. A 21-day fish short-term reproduction assay was conducted in outdoor wetland mesocosms by exposing adult fathead minnows (Pimephales promelas) to graded concentrations of groundwater (1 %, 3 %, and 6 %). Offspring were held in mesocosms up to four days post-hatch to apply a new approach method (NAM), the EcoToxChip™, to explore whether traditional apical endpoints could be predicted using an alternative mechanistic approach. None of the groundwater concentrations used in this study were lethal to fish. There was greater cumulative number of eggs produced at the highest concentration of exposure. However, no abnormal histological appearance was observed in the liver and gonads of fish and no significant effect was observed in the relative expression of genes, tubercle counts, and erythrocyte micronuclei counts compared to the negative control. Food availability in the mesocosms was also assessed and the abundance of zooplankton increased in all groundwater-treated mesocosms. Fathead minnow findings are in contrast to those obtained from previous controlled laboratory studies that revealed significant genotoxicity, hepatotoxicity, and reprotoxicity of the same mixtures. Several factors could explain these observations, including the aging of groundwater in mesocosms before fish addition resulting in photo- and biodegradation and binding to sediments of toxic components. Our static exposure scenario likely underestimated realistic exposure scenarios where groundwater inflow to surface water is generally semi-continuous. Nevertheless, focused transcriptome analysis using EcoToxChips also observed greater toxicity during previous laboratory tests compared to mesocosm scenarios, and thus, our results support the use of this NAM in the ecological risk assessment of contaminated groundwater.

Assessment of sub-lethal effects of Celcron on Java barb through erythrocyte morphology and acetylcholinesterase activity: Implications for environmental health in aquatic ecosystems.

Industrialization and the extensive use of chemicals have raised significant concerns about their environmental impacts, particularly on aquatic ecosystems. This study evaluated the sub-lethal effects of Celcron (Cec), an organophosphate insecticide, on the Java barb (Barbonymus gonionotus) through erythrocyte morphology and acetylcholinesterase (AChE) activity, aiming to refine biomarkers for environmental health assessments. We hypothesized that sub-lethal Cec exposure would induce significant erythrocyte abnormalities and decrease AChE activity in Java barb, with variable recovery rates between gill and kidney tissues. To test this, we exposed the juvenile Java barbs to two sub-lethal Cec concentrations - 0.01 ppm (10 % of the LC50) and 0.05 ppm (50 % of the LC50) -for 60 days. After the exposure period, the fish were placed in pesticide-free water to allow for recovery. Results indicated a significant decline in AChE activity in both liver and kidney tissues, with activity levels showing gradual recovery over time. Erythrocyte abnormalities, including nuclear and cellular changes, were significantly elevated in response to Cec exposure. The frequency of nuclear abnormalities such as micronuclei and binucleation increased in a concentration- and duration-dependent manner, with the gill blood exhibiting higher sensitivity and slower recovery compared to kidney blood. Cellular abnormalities such as twin, teardrop and spindle-shaped cells were also more prevalent in Cec-treated fish. Recovery from these abnormalities was observed but varied between gill and kidney blood, with gill blood showing higher sensitivity and slower recovery compared to kidney blood. This study underscores the utility of AChE activity and erythrocyte abnormalities as biomarkers for assessing pesticide impacts on aquatic organisms. The findings highlight the sensitivity of fish erythrocytes to environmental contaminants and emphasize the need for continued research to better understand the long-term effects of pesticide exposure on aquatic health and ecosystem stability.

Impact of continuous Triazophos exposure on Labeo rohita: Physiological, biochemical, and histological alterations and IBRv2 index assessment.

Pesticides are commonly used in agriculture and aquaculture. Triazophos, an organophosphate-based pesticide, is widely used in agriculture to control many insect pests. Due to its high photochemical stability and mode of action, Triazophos could persist in the aquatic ecosystem and cause toxic effects on non-target organisms. We have studied the potential toxic effects of Triazophos on L. rohita. Primarily, we determined the median lethal concentration (LC50) of Triazophos for 24 and 96 h. Next, we studied acute (96 h, LC50-96 h) toxicity. Then, we studied chronic (35 days, 1/10th LC50-24 h Treatment I: 0.609 mg/L, 1/5th LC50-96 h Treatment II: 1.044 mg/L) toxicity. We analyzed blood biomarkers such as hematology (Hb, Hct, RBC, WBC, MCV, MCH and MCHC), prolactin, cortisol, glucose and protein levels. Concurrently, we analyzed tissue biomarkers such as glycogen, GOT, GPT, LDH and histopathology. IBRv2 index assessment method was also to evaluate the Triazophos toxicity. Studied hematological, hormonal, biochemical and enzymological biomarkers were affected in Triazophos treated groups when compare to the control group. The changes in these biomarkers were statistically significant at the 0.05 alpha level. Triazophos exposed fish shown a severe degenerated primary and secondary lamellae, lamellar fusion, hypertrophy and telangiectasia in the gills. In the hepatic tissue, it caused moderate necrosis, blood congestion, distended sinusoids with minor vacuolation, prominent pyknotic nuclei, hypertrophy, cloudy swelling of cells, lipid accumulation and fibrotic lesions. In the renal tissue, Triazophos caused thickening of Bowman's capsule, hyaline droplets degeneration, irregular renal corpuscle, congestion, cellular swelling, degeneration of tubular epithelium, necrosis, shrunken glomerulus, vacuolated glomerulus, hypertrophy, exudate and edema. IBRv2 analysis suggested that tissue biomarkers are highly sensitive to Triazophos toxicity and prolonged exposure could cause serious health effects like acute toxicity in fish. Triazophos could cause multiorgan toxicity at studied concentrations.

Assessment of fatty acid composition and amino acid profiles of three important fresh water fish species native to river Indus Pakistan.

This study was carried out to analyze the fatty acid and amino acid compositions of three economically important freshwater fish species, Mali (Wallago attu), Raho (Labeo rohita), and Mahseer (Tor putitora), indigenous to Indus River, Pakistan.. Amino acid profiling was done by high-performance liquid chromatography (HPLC) while gas chromatography (GC) was used for fatty acid analysis. Glutamic acid, aspartic acid, arginine, alanine, leucine, lysine and isoleucine were the most predominant amino acid while palmitic acid (C16:0), oleic acid (C18:1c), palmitoliec acid (C16:1c), linolenic acids (C18:2c) and docosahexaenoic acids (DHA C22:6) were the notable fatty acids present in these species. Our results indicated that all species have comparable nutritional composition and are good source of healthy human diet. Our findings will help the people to make informed choice while selecting fish for consumption and will pave the way for future researchers in planning new strategies to enhance the growth and production of commercial fish species. It will also be helpful for theFrom the current finding it was concluded that all fish species under discussion are rich in amino acids and fatty acids. These species contain essential amino acids and important fatty acid such as omega3 and omega 6, thus raising the nutritional quality of these species.

Bioaccumulation of Deltamethrin and Piperonyl butoxide in Labeo rohita fish.

Deltamethrin (DLM), in combination with the synergist piperonyl butoxide (PBO), is extensively used in pest control programs due to its potent pesticidal properties and appreciable safety margin. However, various research studies report their adverse effects on non-target organisms. In this study, we investigated the toxicity of DLM, PBO, and a DLM-PBO (3:1) combination on Labeo rohita (L. rohita) fish fingerlings. Fish behavior and mortality rates were recorded at different time intervals up to 96 h for concentrations of 0.003, 0.007, 0.015, 0.031, and 0.062 µg/mL, respectively. Biochemical, hematological, and histopathological studies were carried out. High-performance liquid chromatography (HPLC) was used to detect and quantify residues in fish samples. The LC50 values after 48 h for DLM, PBO, and DLM-PBO exposed fish fingerlings were found to be 0.028, 0.066, and 0.007 µg/mL, respectively. At a concentration of 0.003 µg/mL of DLM, PBO, and DLM-PBO, the treated fish fingerlings exhibited similar behavior to the control group. Hematological parameters, such as red blood cell (RBC) and white blood cell (WBC) counts, were reduced in the treated groups compared to the control. Biochemical parameters showed increased levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), while total serum protein levels decreased in DLM, PBO, and DLM-PBO treated fingerlings. Histopathological examination of liver, gill, and heart tissues revealed lesions with hydropic degeneration in the liver and fusions of gill lamellae in the treated tissues. Fish fingerlings exposed to the DLM-PBO combination appeared highly prone to toxicity compared to those treated with DLM and PBO separately.

Nano-selenium supplementation: improving growth, digestibility and mineral absorption in freshwater fish, Catla catla.

BACKGROUND: This research investigated the impact of selenium-nanoparticles (Se-NPs) supplemented in sunflower meal (SM)-based diets on digestibility, mineral absorption, and growth performance of Catla catla. Se-NPs were added to seven test meals in varying amounts (0, 0.5, 1, 1.5, 2, 2.5 and 3 mgkg- 1). A total of 315 fish were evenly allocated three replicate tanks, with 15 fish per tank, and given diets equivalent to (5%) of their wet biomass daily for a period of 90 days. RESULTS: The test diet (T-D) supplemented with 1.5 mgkg- 1 Se-NPs yielded remarkable results, including a significant increase in mean weight gain (14 g), an impressive average weight gain percentage (208%), a 100% survival rate, with no mortality observed and the highest specific growth rate (1.25). Moreover, 1.5 mgkg- 1 Se-NPs (T-D-IV) demonstrated superior nutritional digestibility (CP, 76%; CF, 79% and GE, 74%). Additionally, the diet 1.5 mgkg- 1 Se-NPs showed enhanced mineral absorption (K, P, Ca, Na, Zn, Cu and Fe) than other diets. CONCLUSION: It was practically proven that the growth indices, nutrient absorption, and mineral status of C. catla were significantly improved by Se-NPs supplemented SM-based diets, with the optimum level of supplementation being 1.5 mgkg- 1.

New insights into the phylogeny of Carasobarbus Karaman, 1971 (Actinopterygii, Cyprinidae) with the description of three new species.

Fishes from the genus Carasobarbus, widely distributed throughout the river systems of North Africa and West Asia, are commonly referred to as Himris. In the Persian Gulf basin, they are widespread and are also found in fast-flowing rivers or the deeper regions of lakes. In this region, representation of these fishes in scientific collections is scarce, and except for C. luteus, the other species are very poorly documented and frequently misidentified due to their similarities. In this study we analysed the relationships among Carasobarbus species using mitochondrial genes (Cyt b, COI) and present morphological characters based on examinations. Our results revealed three new species which we describe here. Carasobarbus doadrioi, new species, is distinguished by 40-44 scales on the lateral line and a prominent black blotch on end of caudal peduncle in specimens < 85 mm SL. Carasobarbus hajhosseini, new species is distinguished by 32-34 scales on the lateral line and long head length (20-24% SL). Carasobarbus saadatii, new species, is distinguished by 38-40 scales on the lateral line and short head length (19-20% HL). In the Persian Gulf basin, Carasobarbus species exhibit uncorrected genetic distances of 1.6 to 5.5% in the COI barcode region and 2.6% to 9.9% in the Cyt b gene. This study highlights the importance of investigating the unexplored diversity that exists within poorly sampled and understudied freshwater fish group. Such investigations are essential for developing a comprehensive understanding of the true extent of biodiversity, which is critical for informing effective conservation and protection strategies.

Immune response to Aeromonas hydrophila and molecular characterization of polymeric immunoglobulin receptor in juvenile Megalobrama amblycephala.

Polymeric immunoglobulin receptor (pIgR) is an important immune factor in the mucosal immune system of fish, which plays a key role in mediating the secretion and transport of immunoglobulin into mucus. In this study, the full-length cDNA sequence of Megalobrama amblycephala pIgR gene was firstly cloned and the immune response to Aeromonas hydrophila was detected. After being challenged by Aeromonas hydrophila at 3 d, significantly pathological features were observed in intestine, head kidney, spleen, liver and gill of Megalobrama amblycephala. The content of lysozyme (Lys) and the activities of acid phosphatase (ACP) and alkaline phosphatase (AKP) increased significantly at 1 d and reached the peak at 3 d, and the activities of total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-PX) and catalase (CAT) in serum reached the peak at 5 d and 7 d after infection, respectively. The expression level of IL-1ß gene reached the peak at 3 d in intestine, 5 d in gill and spleen, 7 d in head kidney and liver of Megalobrama amblycephala after infected by Aeromonas hydrophila, respectively. The TNF-α gene expression reached the peak at 3 d in intestine and gill, 5 d in head kidney and spleen, 7 d in liver after infection, respectively. The experimental results showed that the infection of Aeromonas hydrophila caused the pathological changes of immune-related tissues and triggered the inflammation responses. The full-length cDNA sequence of Megalobrama amblycephala pIgR was 1828 bp, and its open reading frame (ORF) was 1023 bp, encoding 340 amino acids. The pIgR of Megalobrama amblycephala has a signal peptide sequence, followed by extracellular region, transmembrane region and intracellular region. The extracellular region includes two Ig-like domains (ILDs), and its tertiary structure is twisted "L". The phylogenetic tree was constructed using the adjacency method, and the pIgR genes of Megalobrama amblycephala and cyprinidae fish were clustered into a single branch. Quantitative real-time PCR (qRT-PCR) was used to detect the expression of pIgR gene in different tissues of Megalobrama amblycephala. The expression level of pIgR gene was the highest in liver, followed by intestine, head kidney, skin, middle kidney and spleen, lower in heart, gill and brain, and the lowest in muscle. After being infected by Aeromonas hydrophila, the expression level of Megalobrama amblycephala pIgR gene in intestine, head kidney, spleen, liver and gill showed a trend of increasing first and then decreasing within 28 d. The pIgR gene expression reached the peak in mucosal immune-related tissues (gill and intestine) was earlier than that in systemic immune-related tissues (head kidney and spleen), and the relative expression level of pIgR gene at peak in intestine (12.3 fold) was higher than that in head kidney (3.73 fold) and spleen (7.84 fold). These results suggested that Megalobrama amblycephala pIgR might play an important role in the mucosal immune system to against Aeromonas hydrophila infection.

Molecular cloning and functional characterization of mitochondrial RNA splicing 2 in fish Megalobrama amblycephala, and its potential roles in magnesium homeostasis and mitochondrial function.

Mitochondrial function can be regulated by ion channels. Mitochondrial RNA splicing 2 (Mrs2) is a magnesium ion (Mg2+) channel located in the inner mitochondrial membrane, thereby mediating the Mg2+ influx into the mitochondrial matrix. However, its potential role in regulating the Mg homeostasis and mitochondrial function in aquatic species is still unclear. This study molecularly characterizes the gene encoding Mrs2 in fish M. amblycephala with its functions in maintaining the Mg homeostasis and mitochondrial function verified. The mrs2 gene is 2133 bp long incorporating a 1269 bp open reading frame, which encodes 422 amino acids. The Mrs2 protein includes two transmembrane domains and a conserved tripeptide Gly-Met-Asn, and has a high homology (65.92-97.64%) with those of most vertebrates. The transcript of mrs2 was relatively high in the white muscle, liver and kidney. The inhibition of mrs2 reduces the expressions of Mg2+ influx/efflux-related proteins, mitochondrial Mg content, and the activities of mitochondrial complex I and V in hepatocytes. However, the over-expression of mrs2 increases the expressions of Mg2+ influx/efflux-related proteins, mitochondrial Mg content, and the complex V activity, but decreases the activities of mitochondrial complex III and IV and citrate synthase in hepatocytes. Collectively, Mrs2 is highly conserved among different species, and is prerequisite for maintaining Mg homeostasis and mitochondrial function in fish.

Megalobrama amblycephala IL-22 attenuates Aeromonas hydrophila induced inflammation, apoptosis and tissue injury by regulating the ROS/NLRP3 inflammasome axis.

Mammalian interleukin-22 (IL-22) attenuates organismal injury by inhibiting reactive oxygen species (ROS) and impeding the NLRP3 inflammasome activation. However, the role of fish IL-22 in this process remains unclear. We characterized MaIL-22, an IL-22 homolog in blunt snout bream (Megalobrama amblycephala). Despite its low sequence identity, it shares conserved structures and close evolutionary relationships with other teleost IL-22s. Furthermore, Aeromonas hydrophila (A. hydrophila) infection leads to tissue injury in M. amblycephala immune organs and concomitantly altered Mail-22 mRNA expression, suggesting that MaIL-22 was involved in the antimicrobial immune response. To explore MaIL-22's biological functions, we produced recombinant MaIL-22 (rMaIL-22) protein and demonstrated it significantly enhanced the survival of M. amblycephala post-A. hydrophila infection. To unravel its protective mechanisms, we explored the ROS/NLRP3 inflammasome axis and its downstream signaling responses. The results showed that rMaIL-22 treatment significantly elevated antioxidant enzyme (T-SOD, CAT and GSH-PX) activities to inhibit MDA activity and scavenge ROS in visceral tissues. Meanwhile, rMaIL-22 impeded the activation of NLRP3 inflammasome by suppressing NLRP3 protein and mRNA expression. This indicated that rMaIL-22 contributed to inhibit A. hydrophila-induced activation of the ROS/NLRP3 inflammasome axis. Consistent with these findings, rMaIL-22 treatment attenuated the expression of proinflammatory cytokines (il-1ß, tnf-α and il-6) and proapoptotic genes (caspase-3 and caspase-8) while promoting antiapoptotic genes (bcl-2b and mcl-1a) expression, ultimately mitigating tissue injury in visceral tissues. In conclusion, our research underscores MaIL-22's key role in microbial immune regulation, offering insights for developing IL-22-targeted therapies and breeding programs.

New Insights into the Mechanism by Which the Pituitary Gland Copes with Hypoxia Stress Based on a Transcriptomic Analysis of Megalobrama amblycephala.

Hypoxia is a common environmental stressor in aquatic ecosystems, and during the cultivation process, Megalobrama amblycephala is prone to death because it is hypoxia-intolerant, which brings huge economic losses to farmers. The pituitary gland is a crucial endocrine gland in fish, and it is mainly involved in the secretion, storage, and regulation of hormones. In the present study, we compared the transcriptional responses to serious hypoxia in the pituitary gland among hypoxia-sensitive (HS) and hypoxia-tolerant (HT) M. amblycephala and a control group that received a normal oxygen supply (C0). The fish were categorized according to the time required to lose balance during a hypoxia treatment. A total of 129,251,170 raw reads were obtained. After raw sequence filtering, 43,461,745, 42,609,567, and 42,730,282 clean reads were obtained for the HS, HT, and C0 groups, respectively. A transcriptomic comparison revealed 1234 genes that were differentially expressed in C0 vs. HS, while 1646 differentially expressed genes were obtained for C0 vs. HT. In addition, the results for HS vs. HT showed that 367 upregulated and 41 downregulated differentially expressed genes were obtained for a total of 408 differentially expressed genes. A KEGG analysis of C0 vs. HS, C0 vs. HT, and HS vs. HT identified 315, 322, and 219 enriched pathways, respectively. Similar hypoxia-induced transcription patterns suggested that the downregulated DEGs and enriched pathways were related to pathways of neurodegeneration in multiple diseases, pathways in cancer, thermogenesis, microRNAs in cancer, diabetic cardiomyopathy, and renin secretion. However, in the upregulated DEGs, the PI3K-Akt signaling pathway (C0 vs. HS), microRNAs in cancer (C0 vs. HT), and HIF-1 signaling pathway (HS vs. HT) were significantly enriched. There is a lack of clarity regarding the role of the pituitary gland in hypoxic stress. These results not only provide new insights into the mechanism by which pituitary tissue copes with hypoxia stress in M. amblycephala but also offer a basis for breeding M. amblycephala with hypoxia-resistant traits.

Can Puntius sophore breed artificially under controlled conditions? Tracing the life cycle of Puntius sophore through artificial captive breeding.

The development of new strategies for breeding indigenous fish species is of utmost importance in the wake of unfavorable weather events, as a result of climate change. Therefore, an attempt has been made to achieve artificial breeding of an indigenous barb, Puntius sophore. Two groups of juvenile fish, collected from the wild, were reared till sexual maturation. One group was reared under a natural photothermal regime and the other was reared under strictly controlled conditions with photothermal stimulation till sexual maturation and subsequently, hormonal stimulation with OVAFISH was also done for inducement of spawning. The spawning efficiencies were analyzed and the results in terms of latency period (6.74 Hrs), ovulation rate (92.2 %), fertilization rate (90.6 %), hatching rate (89.9 %), and spawning efficiency coefficient (Se) (0.828) were found better in Puntius sophore reared under the indoor controlled condition with photothermal manipulation and hormone administration compared to the group of fish which was reared under a natural photothermal with a hormonal stimulation. The results of this study demonstrate the captive artificial breeding of Puntius sophore spawners reared under a natural photothermal regime and controlled photothermal regime in indoor conditions. The outcome of the present study can be used for developing key strategies for a climate smart aquaculture for fish farmers.

Comparative Acute Toxicity Studies of Chlorpyrifos Technical Grade with its Emulsifiable Concentrate (20% EC) on Labeo rohita, a Freshwater Major Carp, and Mystus vittatus, a Freshwater Catfish.

Chlorpyrifos is widely used across the world as an organophosphate insecticide and frequently contaminates freshwater bodies through runoff from agricultural fields. In the laboratory, static bioassays were undertaken to examine differences in acute toxicity caused by exposure to the technical grade (94% a.i.) and an emulsifiable concentrate (20% EC) of chlorpyrifos to two species of freshwater fish, Labeo rohita and Mystus vittatus. The recovery of actual chlorpyrifos concentrations varied from 83% (technical grade, T) to 89% (emulsifiable concentrate, F) after two hours in water. The susceptibilities of the two fish species to the two types of chlorpyrifos varied. The 96-h LC50 values for T and F chlorpyrifos in L. rohita were 68 and 36 µg/L, respectively, and 120 and 62 µg/L in M. vittatus, respectively. As the exposure period was extended, the LC50 values gradually decreased. LC50 values between the technical grade and formulation were compared following the criteria of Mayer et al. (1986), Schmuck et al. (1994), APHA (1995), and Demetrio et al. (2014). It was concluded from the study that the emulsifiable concentrate (20% EC) of chlorpyrifos was more toxic than technical-grade chlorpyrifos.

A high-quality chromosome-level genome assembly of the topmouth culter (Culter alburnus Basilewsky, 1855).

Culter alburnus is extensively distributed in various rivers and lakes across China. As a widely adaptive fish species, it has significant economic values and special ecological roles. To meet research demands and provide better genomic resources, in this research, a chromosome-level genome assembly was constructed using HiFi long-reads and Hi-C sequencing data. Compared with the published versions, our genome assembly is of higher quality with only 31 gaps and closer to its true structure and sequence. The genome size was 1.052 Gb, with a contig N50 of 32.92 Mb and a scaffold N50 of 43.09 Mb. 55 contigs were anchored to 24 chromosomes on the basis of Hi-C data. A total of 598.23 Mb of repetitive sequences were annotated and 28,228 protein-coding genes were predicted. Additionally, BUSCO assessment indicated assembly and annotation scores of 98.3% and 99.2%, respectively. This high-quality genome will provide scientific support for excavating the species characteristics of C. alburnus and exploring its molecular mechanisms in response to environmental changes and stress.

Effects of triclosan on lipid metabolism and underlying mechanisms in the cyprinid fish Squalidus argentatus.

The ubiquitous presence of the disinfectant triclosan (TCS) has raised global concerns regarding its potential threat to aquatic organisms. However, the effects of TCS on lipid metabolism in fish and its underlying mechanisms remain unclear. This study investigated the effect of environmentally relevant levels of TCS on the lipid metabolism in the cyprinid fish Squalidus argentatus. Our results showed that the lipid metabolism in the cyprinid fish S. argentatus was perturbed by 28-day exposure to TCS, as evidenced by higher levels of lipid accumulation in both the liver and blood. To elucidate the mechanisms underlying toxicity, we evaluated oxidative stress, inflammatory status, and lipase activity in the liver. Our findings indicated increased ROS-specific fluorescence intensity, superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content in the livers of S. argentatus exposed to TCS, suggesting oxidative damage. Additionally, TCS treatment induced the production of proinflammatory cytokines in the liver of S. argentatus exposed to TCS, which suppressed hepatic lipase activity. Intestinal tissue morphology, inflammation, and blood lipopolysaccharide (LPS) levels were also examined. Significant increases in goblet cell count and MDA levels were observed in the intestinal tract. After 28 days of TCS exposure, the serum LPS levels were significantly elevated. 16S rRNA sequencing was conducted to analyze the effects of TCS on the diversity and composition of the intestinal microbiota. Transcriptomic analysis was performed to reveal global molecular alterations following TCS exposure. In conclusion, our results indicate that TCS may disrupt the lipid metabolism in S. argentatus by (i) inducing hepatic oxidative stress and inflammation, which suppress lipoprotein lipase activity, (ii) affecting the production of beneficial metabolites and endotoxins by dysregulating gut microbiota composition, and (iii) altering the expression levels of lipid metabolism-related pathways.

Assessing spirlin Alburnoides bipunctatus (Bloch, 1782) as an early indicator of climate change and anthropogenic stressors using ecological modeling and machine learning.

Combining single-species ecological modeling with advanced machine learning to investigate the long-term population dynamics of the rheophilic fish spirlin offers a powerful approach to understanding environmental changes and climate shifts in aquatic ecosystems. A new ESHIPPOClim model was developed by integrating climate change assessment into the ESHIPPO model. The model identifies spirlin as a potential early indicator of environmental changes, highlighting the interactive effects of climate change and anthropogenic stressors on fish populations and freshwater ecosystems. The ESHIPPOClim model reveals that 28.57 % of the spirlin's data indicates high resilience and ecological responsiveness, with 34.92 % showing medium-high adaptability, suggesting its substantial ability to withstand environmental stressors. With 36.51 % of the data in medium level and no data in the low category, spirlin may serve as a sentinel species, providing early warnings of environmental stressors before they severely impact other species or ecosystems. The results of uniform manifold approximation and projection (UMAP) and a decision tree show that pollution has the highest impact on the population dynamics of spirlin, followed by annual water temperature, overexploitation, and invasive species. Despite the obtained key drivers, higher abundance, dominance, and frequency values were detected in habitats with higher HIPPO stressors and climate change effects. Integrating state-of-the-art machine learning models has enhanced the predictive power of the ESHIPPOClim model, achieving approximately 90 % accuracy in identifying spirlin as an early indicator of climate change and anthropogenic stressors. The ESHIPPOClim model offers a holistic approach with broad practical applications using a simplified three-point scale, adaptable to various fish species, communities, and regions. The ecological modeling supported with advanced machine learning could serve as a foundation for rapid and cost-effective management of aquatic ecosystems, revealing the adaptability potential of fish species, which is crucial in rapidly changing environments.

Assessment and bioaccumulation of heavy metal contaminants in Golden Mahseer (Tor putitora Hamilton, 1822).

Unpolluted freshwater is a crucial component for maintaining the health of humans. This study aimed to investigate the bioaccumulation and potential health hazards of heavy metal contaminants (Fe, Cd, Cr, Cu, Pb) in water, sediments, and tissues of the golden mahseer fish (Tor putitora) from Zhob River to assess their suitability for human consumption. Samples (soil, water, and fish) were collected from the Zhob River, and Flame Atomic Absorption Spectrometry (FAAS) was employed to measure the concentration of these metals found in soil, water, and various fish body tissues (muscles, skin, gills, and liver). The overall results revealed that water quality parameters, i.e., temperature and pH were found within tolerable ranges, while electrical conductivity and turbidity exceeded the permissible limits of FAO/WHO for fish. Furthermore, this study also identified elevated concentrations of Pb in water and soil, as well as Fe and Cd in soil beyond the standards set by the World Health Organization (WHO) and the United States Environmental Protection Agency (USEPA). In contrast, the concentrations of other targeted metals examined in fish body tissues were found below the permissible limits set by the Food and Agriculture Organization (FAO) and the World Health Organization (WHO), indicating the suitability of this fish species for human consumption. The estimated daily intake (EDI) of these targeted metals in various fish body tissues was found to be within the recommended dietary allowance (RDA), suggesting no associated health risks for the local population. Furthermore, both the Target Hazard Quotient (THQ) and Total Target Hazard Quotient (TTHQ) values measured in this study were less than one, indicating the absence of potential non-carcinogenic health risks related to the consumption of this riverine mahseer fish, but combined metal intake may pose potential health risks. Carcinogenic risk assessment for some metals like Cd, Cr, and Pb revealed no cancer risk for consumers. Moreover, our present research observed comparatively high bioaccumulation (BAF) of each targeted metal in the fish liver from both Zhob River water and soil as compared to other body tissues. Multivariate analysis, including the correlation matrix, revealed strong and significant correlations (P < 0.05) among heavy metal pairs (Fe/Cr, Fe/Pb, Cr/Fe, Cr/Pb, Pb/Fe, Pb/Cr). Hierarchical cluster analysis and Principal Component Analysis (PCA) were utilized to trace the origins of these metals, attributing their presence to nearby rock weathering, mining, as well as municipal and agricultural activities. These factors were recognized as potential sources of heavy metal bioaccumulation in riverine fish. Thus, our current study concluded that the Zhob River was contaminated with these heavy metals and emphasized the need to prevent domestic and industrial sewage inflow. The monitoring of these metals in the food chain was also underscored as crucial for reducing all kinds of associated health risks. This study provides the first report on heavy metal distribution in highly abundant and edible mahseer fishes of the Zhob River.

Comparative mitochondrial genomics analysis of selected species of Schizothoracinae sub family to explore the differences at mitochondrial DNA level.

A comprehensive analysis of the whole mitochondrial genomes of the Schizothoracinae subfamily of the family Cyprinidae has been revealed for the first time. The species analyzed include Schizothorax niger, Schizothorax esocinus, Schizothorax labiatus and Schizothorax plagoistomus. The total mitochondrial DNA (mtDNA) length was determined to be 16585 bp, 16583 bp, 16582 bp and 16576 bp, respectively with 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and 2 non-coding area genes. The combined mean base compositions of the four species were as follows: A: 29.91 % T: 25.47 % G: 17.65 % C 27.01 %. The range of the GC content is 45-44 %, respectively. All protein coding genes (PCGs) commenced with the typical ATG codon, except for the cytochrome c oxidase subunit 1 (COX1) gene with GTG. The analysis of vital amino acid biosynthesis genes (COX1, ATPase 6, ATPase 8) in four different species revealed no significant differences. All 13 PCGs had Ka/Ks ratios that were all lesser than one, demonstrating purifying selection on those molecules. These tRNA genes were predicted to fold into the typical cloverleaf secondary structures with normal base pairing and ranged in size from 66 to 75 nucleotides. Additionally, the phylogenetic tree analysis revealed that S. esocinus species that was most alike to S. labiatus. This study provides critical data for phylogenetic analysis of the Schizothoracinae subfamily, which will help to resolve taxonomic difficulties and identify evolutionary links. Detailed mtDNA data are an invaluable resource for studying genetic diversity, population structure, and gene flow. Understanding genetic makeup can help inform conservation plans, identify unique populations, and track genetic variation to ensure effective preservation.

Physio-metabolic alterations in Labeo rohita (Hamilton, 1822) and native predator Chitala chitala (Hamilton, 1822) in presence of an invasive species Piractus brachypomus (G. Cuvier, 1818).

A 60 days study was conducted to evaluate the physiological response of indigenous species Labeo rohita (LR) and indigenous predator Chitala chitala (CC) in presence of an invasive species Piaractus brachypomus (PB). Two treatment groups as LR + PB (T1) and LR + PB + CC (T2) with individual control groups as T0LR, T0PB and T0CC were designed in triplicates. Fingerlings of LR, PB and CC were randomly distributed into 15 circular tanks with a stocking ratio of 1:1 and 1:1:0.3 in T1 and T2 group, respectively and 10 nos. each of LR, PB and CC in respective control groups. At first 15 min of the experiment, cortisol level was found significantly (P < 0.05) higher in all three experimental fishes in T1 and T2 groups. With the experimental duration, the level of stress hormone (cortisol), oxidative stress enzymes (superoxide dismutase, catalase, and glutathione peroxidase), tissue metabolic enzymes (lactate dehydrogenase and malate dehydrogenase), serum metabolic enzymes (transaminase enzymes) and blood glucose level were significantly (P < 0.05) increased in T1 and T2 groups for LR and CC whereas, no variation (P > 0.05) were observed for PB in both T1 and T2 groups. The total antioxidant capacity (TAC), liver glycogen, total protein, albumin and globulin were found to be significantly (P < 0.05) decreased in LR in the presence of PB and CC. The present study provides a preliminary insight into the biological interaction between native and invasive species and their physiological responses in the presence of native predator with higher trophic index. Thus, the results of the study suggest the superior traits of invasive P. brachypomus try to dominate the other two native species by negatively influencing the native fauna even with a higher trophic index (C. chitala).

Aquatic toxicity and chemical fate of diluted bitumen spills in freshwater under natural weathering.

Increasing global demands for oils are fueling the production of diluted bitumen (DB) from Canada's oil sands region. More weathered than conventional crude (CC) oils, Alberta bitumen is often diluted with lighter petroleum oils to reduce density and viscosity to meet pipeline specifications for transportation. Being a heavy oil product that is transported in large volumes across Canada and the USA, there has been interest to compare its behavior and toxicity characteristics when spilled to those of CC. To determine the influence of environmental weathering upon DB following a freshwater spill, we conducted separate controlled spills of Cold Lake Blend DB and Mixed Sweet Blend light CC oil in a mesocosm spill-tank system at 24 °C with wave-action for 56 days. DB-contaminated waters remained acutely lethal for a period of 14 days to early life stage fathead minnows (Pimephales promelas) exposed during embryologic development, while CC was lethal for 1 day. However, concentrations of mono- and polycyclic aromatic compounds, often claimed to be principally responsible for the acute and chronic toxicity of crude oils, were consistently higher in CC water compared to DB. Elevated aromatic concentrations in CC water correlated with higher prevalences of developmental malformations, reduced heart and growth rates, and impacts on the aryl hydrocarbon receptor pathway. Organic acids were measured over the course of the studies and O2 containing naphthenic acids were present at greater relative abundances in DB- compared to CC-contaminated water, with their attenuation correlating with reduced acute and sublethal toxicity. Furthermore, organic acid degradation products accumulated with time and likely contributed to the consistently sublethal toxicity of the weathered oils throughout the experiment. Improved characterization of the fractions including organic acids and those organic compounds found within the unresolved complex mixture of fresh and weathered crude oils is necessary to adequately understand and prepare for the risks that accidental petroleum spills pose to aquatic resources.