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The present study aimed to elucidate the suitability of formalin and KMnO 4 as therapeutics for fish diseases in Indian major carp, Labeo rohita , while considering their impact on fish stress levels. Acute toxicity tests revealed that the 96-hour LC 50 values for formalin and KMnO 4 were 66.58 ppm and 2.89 ppm, respectively. Sub-lethal concentrations of formalin (6.65 ppm, 3.32 ppm, and 2.21 ppm) and KMnO 4 (0.289 ppm, 0.145 ppm, and 0.096 ppm), along with control groups, were administered to the fish for different exposure periods (24, 48, 72, and 96 hours) and different hematological, biochemical, and immunological parameters were analyzed. The findings demonstrated that formalin exposure resulted in a significant decrease ( p  < 0.05) in hematological parameters, immunological parameters, and serum protein levels. Conversely, formalin exposure led to significant increases ( p  < 0.05) in serum glucose, SGOT, SGPT, and ALP levels. In contrast, KMnO 4 exposure significantly decreased ( p  < 0.05) hematological parameters and serum protein levels, while significantly increasing ( p  < 0.05) immunological parameters. To evaluate curative efficacy, challenge studies were conducted using three sub-lethal concentrations of formalin and KMnO 4 against Aeromonas hydrophila (ATCC 7966) infection. Based on the aforementioned results, the recommended doses of formalin and KMnO 4 were found to be 6.65 ppm and 0.289 ppm, respectively.

This study aimed to assess the toxicity effects of chlorpyrifos and imidacloprid, alone and in combination, on oxidative biomarkers and blood biochemistry of Cyprinus carpio. A total of 324 common carp (Cyprinus carpio) were distributed among 27 tanks and exposed to concentrations of 0.0, 100, and 200 μg L -1 of chlorpyrifos and 0.0, 10.0, and 20.0 μg L -1 of imidacloprid for 28 days. Changes in enzyme activities in the plasma of fish exposed to chlorpyrifos depended on the dose. In contrast, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), creatine phosphokinase (CPK), gamma-glutamyl transferase (GGT) activities were significantly increased in fish exposed to imidacloprid, alone and in combination with chlorpyrifos. However, the activity of butyrylcholinesterase (BChE) was significantly decreased. Exposure to imidacloprid and chlorpyrifos, alone and in combination, increased glucose, urea, cholesterol, triglycerides, and creatinine levels, whereas total protein and albumin levels were significantly decreased. The activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S-transferase (GST), and catalase (CAT) was significantly increased, while glutathione reductase (GR) was significantly decreased. Additionally, although the total antioxidant capacity (TAN) was significantly decreased, malondialdehyde (MDA) levels increased after exposure to imidacloprid and chlorpyrifos, alone and in combination. In conclusion, exposure to imidacloprid and chlorpyrifos, alone and in combination, induced oxidative stress and altered blood biochemistry in carp fish. Moreover, imidacloprid and chlorpyrifos had synergistic effects on some oxidative and biochemical biomarkers., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

To investigate the variations in gene expression in grass carp under high-temperature stress, two libraries were constructed from a high-temperature treatment group (T33) and a control group (T27) and sequenced using Illumina sequencing technology. The results showed that sequencing generated a total of 279,398,348 raw reads, approximately 40.7-51.8 M clean reads were obtained from each library, and the percentage of uniquely mapped transcripts ranged from 80.13 to 84.58%. A total of 260 differentially expressed genes (DEGs) were identified under high-temperature stress, among which 84 genes were upregulated and 176 genes were downregulated. Ten DEGs were randomly selected for quantitative RT-PCR (qRT-PCR) analysis, and the results confirmed that the transcriptome analysis was reliable. Furthermore, the DEGs were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, and the results showed that most of the DEGs were involved in protein, lipid and carbohydrate metabolism. Moreover, plasma urea nitrogen (Urea) and triglyceride (TG) contents were significantly lower in the high-temperature treatment group than in the control group (P < 0.01). In summary, these results indicated that high-temperature stress could inhibit protein synthesis, decrease fatty acid synthesis, and weaken carbohydrate metabolism in juvenile grass carp., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Bisphenol A (BPA) is one of the environmental endocrine disrupting toxicants and is widely used in the industry involving plastics, polycarbonate, and epoxy resins. This study was designed to investigate the toxicological effects of BPA on hematology, serum biochemistry, and histopathology of different organs of common carp ( Cyprinus carpio ). A total of 60 fish were procured and haphazardly divided into four groups. Each experimental group contained 15 fish. The fish retained in group A was kept as the untreated control group. Three levels of BPA 3.0, 4.5, and 6 mg/L were given to groups B, C, and D for 30 days. Result indicated significant reduction in hemoglobin (Hb), lymphocytes, packed cell volume (PCV), red blood cells (RBC), and monocytes in a dose-dependent manner as compared to the control group. However, significantly higher values of leucocytes and neutrophils were observed in the treated groups ( P < 0.05). Results on serum biochemistry revealed that the quantity of glucose, cholesterol, triglycerides, urea, and creatinine levels was significantly high ( P < 0.05). Our study results showed significantly ( P < 0.05) increase level of oxidative stress parameters like reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS) and lower values of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) in treated groups (4.5 mg/L and 6 mg/L)) in the brain, liver, gills, and kidneys. Our study depicted significant changes in erythrocytes (pear shaped erythrocytes, leptocytes, microcytes, spherocytes, erythrocytes with broken, lobed, micronucleus, blabbed, vacuolated nucleus, and nuclear remnants) among treated groups (4.5 mg/L and 6 mg/L). Comet assay showed increased genotoxicity in different tissues including the brain, liver, gills, and kidneys in the treated fish group. Based on the results of our experiment, it can be concluded that the BPA exposure to aquatic environment is responsible for deterioration of fish health, performance leading to dysfunction of multiple vital organs., Competing Interests: All authors declare no potential conflict and no competing interest., (Copyright © 2022 Gulnaz Afzal et al.)

The present study was aimed to evaluate the toxic effects of a commonly used synthetic pyrethroid, λ cyhalothrin on the common carp, Cyprinus carpio L. The results depicted that 96 h LC 50 value of λ cyhalothrin to the fish was 1.48 μg l -1 . During 45 days of chronic exposure a significant reduction (p < 0.05) in the RBC, hemoglobin, and hematocrit value of fish was observed in λ cyhalothrin treated fish. Blood glucose, cholesterol and creatinine levels increased significantly, while total protein and albumin were significantly decreased (p < 0.05) in the exposed fish. Moreover, alanine aminotransferase and aspartate aminotransferase levels in the blood also increased significantly (p < 0.05) in the treated fish. In gills and liver, glutathione S-transferase (GST) and glutathione peroxidase (GPx) and in liver GST exhibited a significant initial augmentation followed by a subsequent reduction while catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA) level increased markedly with incrementing concentrations of λ cyhalothrin in both the organs. Acetylcholinesterase (AchE) activity in both gills and liver decreased in exposed fish upon addition λ cyhalothrin. However, the hazardous effects of λ cyhalothrin on C. carpio were characterized and portrayed by the development of integrated biomarker response (IBR), and biomarker response index (BRI). GUTS-SD and IT modeling were implied for a better interpretation of the toxicity. These results indicate that exposure to λ cyhalothrin alters the survivability at the acute level and the activity of hematological, plasma biochemical as well as enzymological and stress parameters (in gills and liver) at the sublethal level in C. carpio., (Copyright © 2021 Elsevier Inc. All rights reserved.)

This study assessed some important physiological biomarkers of freshwater edible fish Cyprinus carpio following exposure to 10 % (T1) and 20 % (T2) sublethal concentrations of Alpha-cypermethrin (A-cyp) over a total period of 45 days. Behavioral responses were noticed and Kaplan-Meier survival curves were prepared during acute toxicity study. Total serum protein concentration, total erythrocyte count, hemoglobin, packed cell volume, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and total leukocytes count were decreased significantly (p < 0.05), while the blood glucose, total serum lipid concentration, and clotting time were increased significantly (p < 0.05) over control. The most affected fish group and most significantly altered biomarker under toxic stress of A-cyp were identified using integrated biomarker response (IBR). The biomarker response index (BRI) values measured the overall health status of the treated fish and indicated that moderate adverse effects were exerted on the fish group exposed to T2 for 45 days., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Medicinal herbs are used for growth promotion, disease control and other health benefits in aquaculture industry. Here, we examined the effect of dietary laurel-leaf cistus (Cistus laurifolius) ethanolic extract on growth performance, digestive enzyme activity, haematological profile and nonspecific immune responses in common carp (Cyprinus carpio). In addition, resistance against Aeromonas hydrophila infection was examined. Common carp was fed diets containing 0 (Control), 0.1 (CL0.1), 0.5 (CL0.5) and 1 (CL1) g kg -1 laurel-leaf cistus extract for 45 days. After 30 days, superoxide anion production (SAP) increased in CL0.1 and CL0.5 fish groups and at the end of the study all experimental fish groups had higher SAP compared to that of the control (P ˂ 0.05). Lysozyme activity (LA) was elevated in CL0.5 and CL1 treated groups on 30th day (P < 0.05), and this increase was only observed in C0.1 fish group at the end of study compared to control (P ˂ 0.05). Myeloperoxidase activity was significantly increased in CL0.5 and CL1 fish groups at the end of study. IL-1βgene expression was significantly increased in treated fish in a dose-depended manner. Similar results were observed for transcription of IL-6 and IL-8 (P < 0.05). Anti-inflammatory cytokines, IL-10 and TGF-β were highly up-regulated in the intestine and head kidney of CL treated fish groups compared to control (P < 0.05). At the end of experiment, significantly higher final body weight, weight gain, and specific growth rate were obtained in CL0.1 treated fish group compared to control. However, growth was negatively affected in CL1 fish group (P < 0.05). CL1 fish group had also a significantly higher FCR. Amylase activity was significantly increased in all experimental fish groups compared to control (P ˂ 0.05). Trypsin activity was decreased in CL0.1 and CL1 fish groups (P ˂ 0.05). WBC and RBC were significantly increased (P ˂ 0.05) in CL0.5 and CL1 fish groups, whereas haemoglobin, haematocrit, mean cell, mean cell haemoglobin contents were no significantly changed among control and treatment groups. Result of challenge test with A. hydrophila exhibited that survival rate in all treatment groups was significantly higher than that of control. These findings demonstrated that laurel-leaf cistus at 0.1 g kg -1 can be a suitable candidate for growth promotion, immune system induction and infection control in fish., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Selenium (Se), an essential component of deiodinases (DIOs), regulates the contents of thyroid hormones and thus improves animal growth. To explore the influences of selenium supplementation on fish growth metabolism, a total of 270 healthy grass carp (Ctenopharyngodon idella) were divided into three groups and feed three graded dietary selenium (0.141, 0.562, and 1.044 mg Se/kg) levels. The results showed that after 60-day feeding, dietary selenium improved the final body weight and specific growth rate (SGR) of grass carp. The hepatic DIO activities in selenium-supplemented groups were higher than those in control group. A significant increase in triiodothyronine (T3), free triiodothyronine (FT3), and thyroid-stimulating hormone (TSH) levels was accompanied by a decrease in the contents of thyroxine (T4) and free thyroxine (FT4) in selenium-supplemented groups. The histopathological observation of thyroid suggested that selenium deficiency resulted in hypertrophy of follicular epithelial cells. Moreover, the gene relative expression levels of dio1, dio2, and dio3 showed an increasing trend with the rising concentration of dietary selenium. The transcription levels of HPT axis-related genes (crh, tsh-β, ttr, tr-s, tpo, nis) and GH/IGF1-related genes (gh, ghr, igf1, igf1r) were significantly upregulated in selenium-supplemented groups. No significant differences in the above indicators were observed between 0.562 and 1.044 mg Se/kg diet group except T3 content and dio1 relative expression ratio. These results indicate that dietary selenium supplementation improves the hepatic DIO activities and thyroid hormone metabolism and regulates the transcription levels of HPT and GH/IGF axis-related genes, which may be responsible for the growth promotion in grass carp., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Activation of the cAMP pathway by β-adrenergic stimulation and cGMP pathway by activation of guanylate cyclase substantially affects red blood cell (RBC) membrane properties in mammals. However, whether similar mechanisms are involved in RBC regulation of lower vertebrates, especially teleosts, is not elucidated yet. In this study, we evaluated the effects of adenylate cyclase activation by epinephrine and forskolin, guanylate cyclase activation by sodium nitroprusside, and the role of Na + /H + -exchanger in the changes of osmotic fragility and regulatory volume decrease (RVD) response in crucian carp RBCs. Western blot analysis of protein kinase A and protein kinase G substrate phosphorylation revealed that changes in osmotic fragility were regulated via the protein kinase A, but not protein kinase G signaling pathway. At the same time, the RVD response in crucian carp RBCs was not affected either by activation of adenylate or guanylate cyclase. Adenylate cyclase/protein kinase A activation significantly decreased RBC osmotic fragility, i.e., increased cell rigidity. Inhibition of Na + /H + -exchanger by amiloride had no effect on the epinephrine-mediated decrease of RBC osmotic fragility. NO donor SNP did not activate guanylate cyclase, however affected RBCs osmotic fragility by protein kinase G-independent mechanisms. Taken together, our data demonstrated that the cAMP/PKA signaling pathway and NO are involved in the regulation of crucian carp RBC osmotic fragility, but not in RVD response. The authors confirm that the study has no clinical trial., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Kinetoplastid parasites require transferrin (Tf), being the main source of iron, for growth and multiplication. This group of parasites developed a unique receptor-mediated system for acquiring host Tf which bears no structural homology with the host transferrin receptor. Trypanoplasma borreli, a blood parasite of common carp, probably uses a similar mechanism to sequester iron from host transferrin. In this study, we demonstrate a critical role of Tf for parasite growth. For in vitro studies we isolated and purified Tf from carp homozygous for the D or G allele of Tf. We obtained Tf-depleted serum using specific antibodies to carp Tf and studied gene expression in vivo during T. borreli infection with Real Time-quantitative PCR. We demonstrate that T. borreli cannot survive in medium supplemented with Tf-depleted serum while reconstitution with Tf restores normal growth. The critical role of Tf for parasite survival was shown in incomplete medium (medium without serum): addition of purified Tf significantly increased parasite survival. We also demonstrate that Tf polymorphism has a significant impact on T. borreli multiplication. Cultured parasites die more quickly in an environment containing D-typed Tf, as compared to medium with G-typed Tf. Gene expression during T. borreli infection in carp did not show an acute phase response. We could, however, observe an increased transcription of Tf in the head kidney, which may be associated with an immunological function of the Tf protein., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

This study investigated the effect of stocking density on growth performance, blood biochemical profiles, antioxidative capacity, and muscle quality of gibel carp (Carassius gibelio). Gibel carps (initial body weight 57.04 ± 1.89 g) were reared at high stocking density (HSD, 10.85 kg m -3 ), medium stocking density (MSD, 5.06 kg m -3 ), and low stocking density (LSD, 1.47 kg m -3 ) for 60 days. The LSD group exhibited the highest growth rate, while HSD significantly inhibited fish growth. The muscular compositions of crude fat, crude ash, and moisture were significantly changed by stocking density, but crude protein content did not differ significantly. The stocking density altered the muscular texture profiles of gibel carp. Compared to either the HSD group or the MSD group, the highest levels of resilience and springiness occurred in the LSD group. Significant differences were observed in the levels of plasma glucose, alanine aminotransferase, aspartate aminotransferase, cholesterol, and creatinine among three groups. The fish exhibited the highest level of plasma cortisol as well as the lowest levels of triiodothyronine and thyroxine in the HSD group. The fish stocked in the LSD group showed the highest activities of superoxide dismutase, glutathione peroxidase, and catalase as well as the highest content of glutathione in liver. The significant highest total antioxidant capacity occurred in the fish stocked in the LSD group. The results showed that HSD resulted in chronic crowding stress, and exerted negative impact on growth performance, muscle quality, and antioxidative capacity of gibel carp.

The natural antioxidants are well known for their antioxidative activity without side effects when compared to antibiotics. Hence, the present study aimed at evaluating p-Coumaric acid as an antioxidant additive on the blood and mRNA levels of antioxidant-related factors in common carp (Cyprinus carpio). Fish fed the basal diet supplemented with p-Coumaric at 0, 0.5, 1, and 1.5 g/kg for 56 days, then the serum, intestine, and liver samples were collected. The growth performance of fish fed with CA showed significantly (P < 0.05) improved FW, WG, and SGR compared to those of the control one. However, the feed conversion ratio was significantly (P < 0.05) reduced in fish fed 1 and 1.5 g/kg diet levels. SOD was not significantly differed among the groups fed with varied p-Coumaric acid (P > 0.05). Serum GPX and TAC were enhanced considerably by p-Coumaric acid regarding the control with the highest being in fish fed 1.5 g/kg diet (P < 0.05). Serum CAT was more elevated in fish provided p-Coumaric acid at 1 or 1.5 g/kg than the control while fish fed 0.5 g/kg did not display significant changes. MDA level significantly decreased by all p-Coumaric acid groups compared to the control one, and the lowest level was observed in 1.5 g/kg (P < 0.05). The mRNA level of CAT was significantly upregulated in the liver by p-Coumaric acid at 1 or 1.5 g/kg (P < 0.05), while the intestine CAT did not influence by p-Coumaric acid (P > 0.05). The measured SOD in the liver and intestine samples revealed no changes in common carp fed p-Coumaric acid (P > 0.05). GPX was significantly upregulated in the intestine by p-Coumaric acid at 1 or 1.5 g/kg (P < 0.05), whereas the liver GPX was upregulated by p-Coumaric acid at 1.5 g/kg. The mRNA level of the GST gene in the intestine of common carp was upregulated by p-Coumaric acid at 1.5 g/kg, whereas the liver displayed upregulated GST in fish fed 1 g/kg diet. The present study approved the application of p-Coumaric acid as a natural antioxidant for friendly, sustainable aquaculture.

This study aimed to examine the bioavailability (BA) and pharmacokinetic (PK) characteristics of sulfadiazine (SDZ) in grass carp (Ctenopharyngodon idellus) after oral and intravenous administrations. Blood samples were collected at predetermined time points of 0.083, 0.17, 0.5, 1, 2, 4, 8, 16, 24, 48, 72, and 96 hr (n = 6). The samples were extracted and purified by organic reagents and determined by the ultra-performance liquid chromatography. The software named 3P97 was used to calculate relevant PK parameters. The results demonstrated that the concentration-time profile of SDZ was best described by a one-compartmental open model with first-order absorption after a single oral dose. The main PK parameters of the absorption rate constant (K α ), the absorption half-life (t 1/2 Kα ), the elimination rate constant (K e ), the elimination half-life (t 1/2Ke ), and the area under concentration-time profile (AUC 0-∞ ) were 0.3 1/h, 2.29 hr, 0.039 1/h, 17.64 hr, and 855.78 mg.h/L, respectively. Following intravenous administration, the concentration-time curve fitted to a two-compartmental open model without absorption. The primary PK parameters of the distribution rate constant (α), the elimination rate constant (β), the distribution half-life (t 1/2α ), the elimination half-life (t 1/2β ), the apparent distribution volume (V SS ), the total clearance (CL), and AUC 0-∞ were 9.62 1/hr, 0.039 1/hr, 0.072 hr, 17.71 hr, 0.33 L/kg, 0.013 L h -1  kg -1 , and 386.23 mg.h/L, respectively. Finally, the BA was calculated to be 22.16%. Overall, this study will provide some fundamental information on PK properties in the development of a new formulation SDZ in the future and is partially beneficial for the appropriate usage of SDZ in aquaculture., (© 2020 John Wiley & Sons Ltd.)

The present study aimed to investigate the potential effects of dietary marjoram extract on growth performance, health, and disease resistance in common carp, Cyprinus carpio. To this purpose, the fish were assigned into four treatments and fed by diets supplemented with 0 (control), 100, 200, and 400 mg marjoram extract kg -1 over eight weeks and then challenged with Aeromonas hydrophila. According to the results, 200 mg kg -1 dietary marjoram extract inclusion showed the highest final weight, weight gain, and specific growth rate, and lowest feed conversion ratio (FCR). White blood cell number, Red blood cells, hematocrit, hemoglobin, mean corpuscular volume and mean corpuscular hemoglobin were markedly increased particularly at 200 mg kg -1 treatment. Marjoram extract significantly increased plasma superoxide dismutase activity and decreased malondialdehyde level compared to the control treatment. Plasma complement and lysozyme activities and total immunoglobulin levels, mucosal complement, lysozyme and alkaline phosphatase activities and immunoglobulin levels were significantly increased compared to the control group. The lowest post-challenge survival rate was observed in the control treatment, whereas the highest value was related to the 200 mg kg -1 marjoram treatment. In conclusion, the present study demonstrated that, marjoram extract is a suitable feed supplements for common carp, as it stimulates the fish growth, antioxidant, and immune systems, which suppress the fish mortality during Aeromonas septicemia. According to the results, 200 mg marjoram extract kg -1 is recommended for carp feed formulation., (Copyright © 2020. Published by Elsevier Ltd.)

Naked carp (Gymnocypris przewalskii), endemic to the saline-alkaline Lake Qinghai, have the capacity to tolerate combined hypoxia and high alkalinity. This study evaluated the effect of the interaction between carbonate alkalinity and hypoxia on ammonia excretion and blood gas variation in naked carp. Naked carp were subjected to normoxic, hypoxic and reoxygenation phases at two different carbonate alkalinity levels (CA0 = 0 mmol/L; CA32 = 32 mmol/L) for 4 days. The ammonia excretion rate (J Amm ) of the CA0 group rapidly decreased under hypoxia and recovered under normoxia for four consecutive days. The J Amm under CA32 also decreased under hypoxia and recovered to its previous level in the first 2 days. However, the J Amm under CA32 was lower than that under CA0. The blood pO 2 , sO 2 of CA0 and CA32 group was significantly reduced under hypoxia, after which both groups recovered. Blood pCO 2 of the CA32 group was lower than CA0 throughout the experiment. There were no changes in haematocrit of the naked carp exposed to carbonate alkalinity and hypoxia. The alkaline water increased the pH of the blood and contributed to increased haemoglobin O 2 affinity. Overall, the present findings reveal that naked carp is a tolerant species that can maintain main ionic homeostasis under severe alkalinity and hypoxia. The high alkaline water is beneficial for naked carp to adapt to hypoxic environment.

A 56-day research was performed to examine the influence of graded levels (0 (control), 0.5, 1 and 2%) of Fern (Adiantum capillus-veneris) leaves powder (FLP) in diet on immune competence and growth of common carp (Cyprinus carpio, initial weight = 20 g). The serum total immunoglobulins content and lysozyme activity in the 1 and 2% FLP groups remarkably increased compared to the other groups (P < 0.05). The skin mucosal lysozyme activity enhanced with increasing dietary FLP level in a dose-response manner. Fish fed on the FLP-supplemented diets had higher skin-mucosal superoxide dismutase activity than the control (P < 0.05). However, serum antioxidant enzymes were not affected by dietary fern (P > 0.05). The serum bactericidal activity against human and fish pathogens increased with enhancing the FLP level in diet against Staphylococcus aureus, Escherichia coli (EHEC ATCC 43895), Escherichia coli (CI), Pseudomonas aeruginosa, Klebsiella pneumonia and Aeromonas hydrophila. The serum antibacterial activity against Yersinia ruckeri in the 2% FLP group was higher than the other treatments. Furthermore, the serum bactericidal activity against P. aeruginosa (ATCC 27853) only observed in fish fed on the 1 and 2% FLP-supplemented diets. The skin mucosal bactericidal activity and inhibitory effects increased with enhancing the FLP level in diet against E. coli, K. pneumonia, Y. ruckeri and A. hydrophila in a dose response manner. Moreover, the skin mucosal bactericidal activity against S. aureus only observed in fish fed on 1 and 2% FLP-supplemented diets. The weight gain values in the 1 and 2% FLP groups were higher than the other treatments (P < 0.05). Feed conversion ratio (FCR) improved with increasing FLP level in diet in a dose-response manner (P < 0.05). By considering serum and mucosal bactericidal activities against different pathogenic bacteria, the supplementation of 2% FLP in diet is recommended for C. carpio during grow-out phase., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The study was designed to evaluate the possible protective roles of dietary Origanum vulgare essential oil (OVEO) against cypermethrin (CP)-induced serum biochemical changes and oxidative stress of common carp (Cyprinus carpio). Moreover, histopathological alterations, apoptosis, cell proliferation, and DNA damage in the gills and hepatic tissues were also assessed. Briefly, fish were allotted into six groups with three triplicates whereas a group fed on basal diet and did not exposed to CP and served as control (CTR), two groups were fed on diets supplemented with two levels of OVEO (0.5 % and 1.0 %), a group exposed to sub-lethal concentration of CP (1/10 of 96 h-LC 50 = 0.4134 μg/L), and two other groups exposed to the same concentration of CP and fed on diets supplemented with both levels of OVEO (CP + 0.5 % OVEO, and CP + 1.0 % OVEO), respectively, for 30 days. CP induced significant elevation of serum alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), urea, and creatinine levels indicating hepato-renal toxicity (P < 0.05). Besides, there was a significant decrease in serum catalase (CAT) and superoxide dismutase (SOD) activities (P < 0.05). Moreover, CP induced significant histopathologic alterations in gills, anterior kidneys, and hepatic tissues with activation of apoptosis (Caspase-3) and proliferating cell nuclear antigen (PCNA). Comet assay demonstrated significant DNA damage in gills and liver tissues of the CP-exposed group. Interestingly, a significant attenuation of serum ALT, AST, ALP, urea, creatinine, CAT, and SOD levels (P < 0.05) was noticed in CP-exposed fish and concurrently fed diets supplemented with either 0.5 % or 1.0 % OVEO. Moreover, histopathologic alterations and apoptosis were significantly reduced along with a concomitant significant decrease in DNA damage (P < 0.05) which indicated the mitigation of DNA damage. Conclusively, the study showed that OVEO is an effective counteractive treatment against CP-induced damage in exposed common carp and is recommended during the formulation of fish rations., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Increase in water temperature due to anthropogenic and climatic changes is expected to affect physiological functions of fish. In this study, we determined high temperature tolerance (CTmax) of a common aquacultured Indian major carp, rohu, Labeo rohita fingerlings (15.96 ± 0.72 g BW, 11.56 ± 0.42 cm TL) followed by acclimatization at three temperatures (30, 33, 36 °C). To determine the CTmax, we analyzed the major hemato-biochemical indices - hemoglobin (Hb), red blood cell (RBC), white blood cell (WBC), blood glucose levels, and erythrocytic nuclear abnormalities (ENAs) and erythrocytic cellular abnormalities (ECAs) of peripheral erythrocytes in the fish sampled at the start and end point at each acclimated temperature. Significantly decreased CTmax of the fish was found at 36 °C compared to 30 °C and 33 °C. The fish in the highest (36 °C) temperature were found with significantly lower Hb and RBC content and significantly higher WBC and blood glucose levels than that of the fishes in the lowest (30 °C) temperature both at the start and end points. The highest frequencies of ENAs and ECAs were found in the highest (36 °C) temperature group compared to the lowest (30 °C) temperature group at both the points. We also evaluated growth performance of the rohu fingerlings reared in the three temperatures for 60 days. The growth parameters - final weight gain, percent weight gain and specific growth rate were the highest at 33 °C and the lowest at 36 °C. The present study revealed that the highest temperature (36 °C) tested here may be hazardous to rohu and the temperature should be kept below 36 °C in the aquaculture setting to avoid physiological damage and growth and production loss to the fish., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Spring viremia of carp virus (SVCV) causes devastating disease in aquaculture, resulting in significant economic impact. To develop an effective means against SVCV infection, a Lactococcus lactis (L.lactis) based subunit vaccine (pNZ-UGA) was developed based on surface displaying of SVCV glycoprotein using anchoring motif of the cA (C terminus of the peptidoglyvsn-binding) domains of AcmA, a major autolysin from L.lactis. The surface expression of SVCV glycoprotein was verified by indirect immunofluorescence assay. The efficacy of the constructed vaccine was further evaluated in common carp. The results showed that the higher levels of specific IgM could be detected in fish vaccinated with pNZ-UGA, compared with that in PBS and L.lactis groups. Immune-related genes including TNF-α, IL-6b, IL-1β, Cxcr 1, Cxca, IFNg2b, I-IFN, and IgM expression in pNZ-UGA group were strongly up-regulated, revealing that robust innate immune response was induced. Notably, the lowest cumulative mortality (13.46%) was observed in fish vaccinated with pNZ-UGA vaccine after SVCV challenge, whereas the cumulative mortality were 100.00% and 92.31% in PBS and L.lactis groups, respectively. This study suggests the potential use of the recombinant L.lactis with surface displaying antigen proteins as effective vaccines against SVCV and other fish virus infection., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

This study was performed to determine the effects of pectin derived from orange peel (PDOP) on growth performance, antioxidant enzyme activity and serum and skin mucus immune response of common carp (Cyprinus carpio). Common Carp (16.94 ± 0.03 g) were distributed into 12 tanks representing four treatments repeated in triplicates. Four diets were prepared to contain four levels of PDOP as follows: 0 (control), 0.5, 1, and 2% PDOP. Growth and immunological parameters as skin mucus lysozyme activity (SMLA) and total immunoglobulin (SMTIg), serum total immunoglobulin (STIg), serum peroxidase activities (SPA), Catalyse activity (CAT), DPPH radical scavenging activity, specific growth rate (SGR), weight gain (WG), final weight (FW), and feed conversion ratio (FCR) were assessed. Fish fed diets supplemented with PDOP showed an improvement of SGR, WG, FW, and FCR (P < 0.05). In terms of skin mucus immunological parameters, dietary inclusion of pectin significantly (P < 0.05) increased SMTIg. Likewise, carps fed either 1 or 2% PDOP showed notable enhancement of SMLA. In the case of serum immune parameters and antioxidant defence, carps in 1% PDOP treatment showed significantly (P < 0.05) higher SPA and CAT compared to fish fed either control diet or 0.5% OPDP. Additionally, no significant change (P > 0.05) was found in SPA and CAT of fish fed either 1% PDOP or 2% PDOP. Also, no significant (P > 0.05) difference was noticed between treated groups and control in the case of STIg. Furthermore, no significant differences were observed in DPPH radical activity among treatments (P > 0.05). Overall, these results suggested that inclusion of PDOP in common carp diet can beneficially affect growth and immune response., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Cadmium (Cd), a substance with one of the most critical health hazard indices, can cause damage to both the blood and kidneys and accumulates in the body at last. The present work studied the toxicological effects of Cd and the therapeutic effects of metallothionein (MT) and vitamin E (VE) on the trunk kidney and blood of freshwater grass carp (Ctenopharyngodon idellus). Grass carp were divided into three groups: Cd + phosphate-buffered saline (PBS) group, Cd + VE group, and the Cd + MT group. Fish were injected with CdCl 2 on the first day and then VE, MT, or PBS was administered 4 days post-injection. Fish not injected with Cd were used as a negative control. The blood and trunk kidney amassed Cd and suffered severe damage in the forms of organ toxicity cytotoxicity, and immunotoxicity. However, the MT reduced the Cd content in the trunk kidney and blood and partially stabilized the damaged organs. Treatment with VE, however, only demonstrated weaker protection against on Cd-induced toxicity. The results indicate that exogenous MT may play an essential role in restoring homeostasis of the Cd-poisoned urinary and circulatory system and that it may help eliminate Cd in aquatic animals.

Demand for omega-3 long chain polyunsaturated fatty acids has become global challenge for aquaculture and different components have been used to increase nutritional value of fillets. The aim of this study was to evaluate influences of feeds on zootechnical parameters, biochemical plasma parameters, expression of lipid-dependent genes, hepatocyte histomorphologies, and fatty acid profiles in common carp fillets. We compared a control diet (CTRL), mimicking a commercial feed formulation for common carp, with three diets containing blends of vegetable oils and a DHA-rich alga (Schizochytrium sp.) included at 3.125% (CB1) or 1.563% (CB2), and 2.1% salmon oil (CB3). The study revealed no differences in final body weight of fish fed CB1-3 diets in comparison with significantly lower CTRL. Concentrations of all biochemical parameters in plasma increased gradually in fish fed CB1-3 diets when compared to CTRL diet, with exception of triacylglycerol levels. Expression of hepatic fas, elovl-5a and pparα genes increased significantly in fish fed CB1 and CB2. Additionally, eicosapentaenoic (EPA) and docosahexaenoic acid (DHA) accumulation in muscle tissue was directly proportional to the amounts supplied in the diets. Our study revealed that carp fillet profiles can be manipulated for DHA and EPA-contents using enriched diets, depending on the source of fat., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The initiation of eryptosis as a result of genotoxic action of Cr(VI), seen through micronucleus and comet assay in the peripheral erythrocytes of Ctenopharyngodon idellus was evaluated through RT-qPCR. For this, fish was exposed to sublethal concentration of hexavalent chromium (5.30 and 10.63 mg/L), and the blood was sampled on different endpoints (15, 30 and 45 days). Accumulation of chromium in the erythrocytes was also studied, which depicted a significant increase in toxicant concentration and time dependent manner. Both concentrations of hexavalent chromium induced DNA damage, visible in the form of comet tails. The presence of micronuclei in the erythrocytes was accompanied with occurrence of nuclear bud (NBu), lobed nucleus (Lb), notched nucleus (Nt), vacuolated nucleus (Vn), binucleated cell (Bn) as nuclear abnormalities; and acanthocytes (Ac), echinocytes (Ec), notched cells (Nc), microcytes (Mc) and vacuolated cytoplasm (Vc) as cytoplasmic abnormalities. The expression of genes related to intrinsic apoptotic pathway induced by Cr(VI) presented significant (p < 0.05) upregulation in the expression of p53, Bax, Apaf-1, caspase9 and caspase3, and downregulation of Bcl2; inferring the initiation of apoptotic pathway. The ration of Bax and Bcl2 also appended the apoptotic state of the erythrocytes. From the present investigation, it can be concluded that genotoxicity induced by hexavalent chromium lead to eryptosis in C. idellus., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Fish erythrocytes are very sensitive to environmental pollution because they exhibit pathological alterations before appearance of other external symptoms, may be used as reliable toxicity bioindicators. Microscopic analysis can be a useful tool in early detection of alterations in erythrocyte morphology due to different environmental stressors, and in predicting organism's possible response ways. This study aims to evaluate the toxicity of a polluted freshwater lake on the erythrocytes of crucian carp fish, Carassius carassius, using digital light microscopy (DLM). Scanning DLM micrographs revealed the presence of nuclear and cytoplasmic abnormalities like micronucleus, nuclear buds, lobed nucleus, deformed nucleus, vacuolated cytoplasm, and echinocyte in the erythrocytes of crucian carp fish from a polluted lake. The obtained results substantiate that computation of erythrocyte morphological damages as biomarkers of exposure, through digital light microscopy software, can be effectively used as a toxicological tool., (© 2019 Wiley Periodicals, Inc.)

The present study was designed to investigate the dietary effects of ginger extract (Zingiber officinale) on common carp (Cyprinus carpio). Three hundred and sixty fish weighing 10.9 ± 0.17 g were randomly divided into four experimental treatments in triplicates. Four experimental diets were designed containing different inclusion levels of ginger extract as 0% (control), 0.1%, 0.2%, and 0.4% named T1, T2, T3, and T4, respectively, and fish were fed 3% of body weight daily for 60 days. Feed conversion ratio, weight gain, and specific growth rate parameters were measured higher in the groups fed with ginger extract compared to the control, especially in T3 (P < 0.05). Also, fish treated with ginger extract showed lower carcass moisture and lipid, but higher protein contents compared to the control. The results showed significant increase in the number of erythrocytes and leucocytes, as well as the level of hematocrit and hemoglobin in fish fed with supplemented diets. In addition, significant enhancement was observed in cases of serum parameters, including total protein, albumin, globulin, lysozyme, superoxide dismutase, catalase, and total immunoglobulins in T3 and T4 compared to the control. The lowest serum cortisol level was noticed in T3. Furthermore, the T3 group showed the highest activities of skin mucus lysozyme, alkaline phosphatase, and protease (P < 0.05). In conclusion, the present results demonstrated that Zingiber officinale extract at a 0.2% level can effectively improve the growth and health status of Common Carp (Cyprinus carpio)., Competing Interests: Declaration of competing interest There are no conflicts to declare., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Anesthesia is commonly employed in aquatic medicine to facilitate physical exams, diagnostics, and surgical interventions. Tricaine methanesulfonate (MS-222) is the most commonly used anesthetic for fish and is currently the only anesthetic approved by the US Food and Drug Administration Center for Veterinary Medicine for food-producing fish. Despite the frequency of anesthetic procedures in fish, anesthetic monitoring remains rudimentary in many facilities. This study evaluated the impact on blood gases, acid-base balance, and electrolytes in koi ( Cyprinus carpio ) anesthetized at concentrations of 100 mg/L and 150 mg/L MS-222. Blood samples from 25 fish per treatment were collected at 5 and 20 min of anesthetic immersion. Forty-nine of 50 fish recovered uneventfully from anesthesia; one fish did not recover and was euthanatized. Results showed significant increases in partial pressure of carbon dioxide (pCO 2 ) ( P = 0.006) and hyperglycemia ( P = <0.0001) with increasing anesthetic concentration and time under anesthesia and a significant decrease in partial pressure of oxygen (pO 2 ) with increased anesthetic time ( P = 0.021). There were several electrolyte changes observed with both increasing anesthetic time and concentration. All electrolytes except potassium remained within published reference ranges for koi, while potassium showed a significant decrease in concentration associated with anesthetic time and concentration. The results of this study indicate that MS-222 at 100 mg/L and 150 mg/L represent safe anesthetic concentrations for koi undergoing minimally invasive diagnostics; however, koi anesthetized with MS-222 at a concentration of 150 mg/L experienced more significant changes in blood gases, acid-base balance, and electrolyte concentrations.

Reproduction is a major component of an animal's life history strategy. Species with plasticity in their reproductive biology are likely to be successful as an invasive species, as they can adapt their reproductive effort during various phases of a biological invasion. Silver carp (Hypophthalmicthys molitrix), an invasive cyprinid in North America, display wide variation in reproductive strategies across both their native and introduced ranges, though the specifics of silver carp reproduction in the Illinois River have not been established. We assessed reproductive status using histological and endocrinological methods in silver carp between April and October 2018, with additional histological data from August to October 2017. Here, we show that female silver carp are batch spawners with asynchronous, indeterminate oocyte recruitment, while male silver carp utilize a determinate pattern of spermatogenesis which ceases in the early summer. High plasma testosterone levels in females could be responsible for regulating oocyte development. Our results suggest that silver carp have high spawning activity in the early summer (May-June), but outside of the peak spawning period, female silver carp can maintain spawning-capable status by adjusting rates of gametogenesis and atresia in response to environmental conditions, while males regress their gonads as early as July. The results of this study are compared to reports of silver carp reproduction in other North American rivers as well as in Asia., (© The Author(s) 2019. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objective: Assessment of the effect of additional acute irradiation (AAI) in sub-lethal range on the peculiarities of formation of qualitative composition of red blood and adaptive possibilities of the hematopoietic system of fish in reservoirs within the Chernobyl Exclusion Zone (CEZ), contaminated with radionuclides., Materials and Methods: We studied the Prussian carp Carassius gibelio Bloch (2-3 years old) from Vershyna Lake (impacted reservoir in the CEZ) and Dibrova Lake (reference reservoir), where the average absorbed dose rate (ADR) was, respectively, 110.80 and 0.07 μGy/h. The fish were exposed by a single X-ray dose at the unit RUM-17 (ADR - 0.89 Gy/min) at dose of 2.5, 5.0 and 10.0 Gy. Morphological and cytogenetic studies of red blood cells were per- formed dynamically for the 1st, 7th and 30th day after irradiation. The blood was taken from the tail vein and stained by Pappenheim's method. Cytomorphological abnormalities of erythrocytes were analyzed for 3000 cells (‰)., Results: In blood of fish from the impacted reservoir (Vershyna Lake) on the 1st day after AAI, the average number of all types of deformations of erythrocytes cells increased in 2.1 times, and in fish of the reference reservoir (Dibrova Lake) - in 5.2 times. General spectrum of erythrocyte disturbances in fish of Vershyna Lake has increased from 4 to 6 types (in 1.5 times), and Dibrova Lake - from 2 to 5 types (in 2.5 times). However, further observations of the dynamics of morphological disorders of fish's blood cells from Vershyna Lake on the 7th and the 30th day after AAI showed a significant increase in pathological processes in comparison with the reference reservoir, namely - an increase in the number of gross damage of DNA molecules - amitosis (on average in 14.8 times), erythrocytes with septum in the nucleus (in 1.9 times), as well as structural disorders in the cells - protuberances (in 10.5 times), cells with vacuolate cytoplasm (in 8.6 times)., Conclusions: The red blood cells of the fish of the impacted reservoir show a greater vulnerability to AAI, which manifests itself in an increase in the number of erythrocytes with pathology of mitosis and may indicate a violation of the genetic structures of cells due to long-term radiation exposure., (N. A. Pomortseva, D. І. Gudkov.)

Rise of water temperature as a consequence of global warming is anticipated to affect the physiological activities of fish, especially in tropical regions. In the present experiment, we exposed the Indian major carp, rohu Labeo rohita to three different temperature regimes (30 °C as control and 33 °C and 36 °C) for 60 days and observed the effects of these temperature on: major hemato-biochemical indices (Hemoglobin; Hb, Red blood cell; RBC, White blood cell; WBC and blood glucose levels), erythrocytic nuclear abnormalities (ENA), and erythrocytic cellular abnormalities (ECA) of peripheral erythrocytes along with the formation of differential leucocytes in the blood. Fish were sacrificed at day 7, 15, 30 and 60 after the start of exposure to the temperature regimes. Hb decreased significantly on days 7 and 15 at 36 °C. Throughout the study period, the decrease of RBC and increase of WBC were significant at 36 °C. Blood glucose level increased significantly initially at day 7 but decreased significantly at day 60 at 36 °C. Frequencies of ENA (binucleated, nuclear bud, nuclear bridge, karyopyknosis and notched nuclei) and ECA (twin, fusion, echinocytic, spindle, tear drop and elongated shaped) were significantly increased at the highest temperature (36 °C) at almost all of the sampling days. In the case of differential leucocyte count, high temperature caused a significant increase in the number of neutrophils and a significant decrease in the number of lymphocytes. Overall, these results indicate that chronic exposure to high temperature (36 °C) induces a number of stress responses in rohu and that temperature should be kept below 36 °C in the aquaculture setting to avoid damage to the fish., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The aim of this work is to evaluate the effects of dietary inclusion of mixed plant protein (MP) (rapeseed meal:cottonseed meal:peanut meal = 1:1:1) on growth, body composition, blood biochemical parameters, growth hormone/insulin-like growth factor 1, and relative non-specific immune response in Yellow River carp Cyprinus carpio. Five isonitrogenous and isoenergetic trial diets were formulated to replace fish meal at 0 (MP0, control), 25% (MP25), 50% (MP50), 75% (MP75), and 100% (MP100) mixed plant protein, respectively. The 25% mixed plant protein did not affect the weight gain, specific growth rate, and protein efficiency ratio, whereas these parameters were depressed by 50% and above mixed plant protein. The whole body protein content gradually decreased with increasing dietary MP; meanwhile, the whole body lipid content is the opposite. The MP75 and MP100 diets adversely affected the glucose level, total cholesterol value, alanine transaminase, and aspartate transaminase activity of serum. Fish fed MP75 and MP100 diets showed higher growth hormone level than that of MP0 diet; however, the insulin-like growth factor 1 level got the opposite result. The 50% and above inclusion of MP decreased lysozyme activity and increased malondialdehyde content. In conclusion, no more than 50% of fish meal could be replaced by mixed plant protein in diet. However, 50% and above inclusion of mixed plant protein in diet could depress the growth, insulin-like growth factor 1 level, and non-specific immune response, and significantly affect the whole body composition and serum biochemical parameters in Yellow River carp.

The grass carp (Ctenopharyngodon idella), blunt snout bream (Megalobrama amblycephala) and yellow catfish (Pelteobagrus fulvidraco) are economically important fishes in China. Fish hematological features, especially the type and number of peripheral blood cells, are crucial for the evaluation of fish health and the diagnosis of fish diseases. Since the automatic blood cell count equipment for human is not suitable for fishes, the manual method is critical in the quantification of fish blood cells. To make sense of the comparison and interpretation of the blood cell count studies in different articles, the standardization of blood cell classification is necessary. In this study, erythrocytes (red blood cell, RBC), thrombocytes (TC) and leucocytes (i.e. white blood cells, WBC, including lymphocytes, neutrophils and monocytes) were well distinguished in blood smears with Giemsa staining and confirmed by transmission electron microscopy. RBC, TC and WBC were directly counted with an improved Neubauer counting chamber in a modified diluting solution. The differential leucocyte count (DLC) was carried out in blood smears. In view of the labeling characteristics of peroxidase (PO) positivity in neutrophils and non-specific esterase (α-ANAE) positivity in monocytes, PO positive cell percentage and α-ANAE positive cell percentage were also determined in cytochemistry staining smears. No difference was found for the percentages of neutrophils and monocytes between Giemsa staining and cytochemistry staining. The standardized classification, normal count ranges and sizes of the peripheral blood cells by the present systemic studies will provide useful references for monitoring the health status of grass carp, blunt snout bream and yellow catfish., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The present study investigated the effects of dietary olive leaf (Olea europea L.) extract (OLE) on some blood parameters and immune (TNF-α, IL-1ß and IL-8) related genes in different tissues (head kidney, liver and spleen) and resistance of common carp, Cyprinus carpio to Edwardsiella tarda. Five diets were prepared for fed fish (mean body weight 15.90 ± 0.93 g) with different rates of OLE (0.0%, 0.1%, 0.25%, 0.50% and 1.0%). A control diet was prepared non-supplemented with OLE. Increased haematocrit ratio, serum myeloperoxidase activity, immune response gene levels (IL-1β in head kidney tissue and TNF-α in spleen tissue) and survival rate against E. tarda particularly in the 0.1% OLE treatment group. In conclusion, results of the present study show that feeding common carp with a diet containing 1 g/kg OLE over a period of 60 days might be adequate to improve fish immune parameters, and survival rate against E. tarda. Therefore, OLE can be used as a dietary additive to prevent E. tarda in common carp., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Serine peptidases are involved in many physiological processes including digestion, haemostasis and complement cascade. Parasites regulate activities of host serine peptidases to their own benefit, employing various inhibitors, many of which belong to the Kunitz-type protein family. In this study, we confirmed the presence of potential anticoagulants in protein extracts of the haematophagous monogenean Eudiplozoon nipponicum which parasitizes the common carp. We then focused on a Kunitz protein (EnKT1) discovered in the E. nipponicum transcriptome, which structurally resembles textilinin-1, an antihemorrhagic snake venom factor from Pseudonaja textilis. The protein was recombinantly expressed, purified and biochemically characterised. The recombinant EnKT1 did inhibit in vitro activity of Factor Xa of the coagulation cascade, but exhibited a higher activity against plasmin and plasma kallikrein, which participate in fibrinolysis, production of kinins, and complement activation. Anti-coagulation properties of EnKT1 based on the inhibition of Factor Xa were confirmed by thromboelastography, but no effect on fibrinolysis was observed. Moreover, we discovered that EnKT1 significantly impairs the function of fish complement, possibly by inhibiting plasmin or Factor Xa which can act as a C3 and C5 convertase. We localised Enkt1 transcripts and protein within haematin digestive cells of the parasite by RNA in situ hybridisation and immunohistochemistry, respectively. Based on these results, we suggest that the secretory Kunitz protein of E. nipponicum has a dual function. In particular, it impairs both haemostasis and complement activation in vitro, and thus might facilitate digestion of a host's blood and protect a parasite's gastrodermis from damage by the complement. This study presents, to our knowledge, the first characterisation of a Kunitz protein from monogeneans and the first example of a parasite Kunitz inhibitor that impairs the function of the complement., (Copyright © 2019 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Interactions between trace metals, serum biochemical parameters, and oxidative status markers were observed. Freshwater fish Cyprinuscarpio blood samples (n = 38) were collected at the beginning of May (n = 19) and at the end of July (n = 19) of 2015. The concentrations of metals (As, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Se, Sr, and Zn) were analyzed in blood serum samples of fishes by inductively coupled plasma optical emission spectrometry (ICP-OES), and Hg was determined by cold-vapor atomic absorption spectroscopy (CV-AAS). The general scheme of descending concentrations of metals in blood serum samples was as follows: Zn > Fe > Cu > Sr > Cr > Ni > Mn > Pb > Se > As > Cd > Hg. Zn was the most accumulated element (4.42-119.64 mg/L) in both seasons. Overall, the trace element content was higher in spring season, except Hg, Ni, Se, and Sr. The seasonal effect was confirmed for Mn, Zn, Mg, Glu, AST, and Chol levels and for most oxidative status markers. The gender effect was confirmed for Sr, GPx, PC, Chol, and CK concentrations. Trace metals (especially Cd, Cr, Cu, Fe, Hg, Mn, Ni, Sr, Zn, As) significantly affected some blood serum chemistry parameters. The correlation analysis between oxidative status markers (ROS, TAC, MDA, SOD, GSH, UA, BHB, and Alb) and trace metal (Cd, Cu, Ni, Sr, Hg, Pb, Fe, Mn) content confirmed statistically significant interactions in both seasons. Obtained results indicate specific actions of trace metals.

In the present study, sub-lethal effects of chlorpyrifos, alone (25 and 50 μg L -1 ) and in combination with Polyethylene glycol (5 and 10 mg L -1 ), on common carp were investigated. Exposure to chlorpyrifos caused significant changes in all blood biochemical parameters compared to control groups. Total protein, triglyceride, glucose levels, as well as lactate dehydrogenase (LDH), gamma-glutamyl transferase (GGT), and creatine phosphokinase (CPK) activities in plasma changed after exposure to 10 mg L -1 polyethylene glycol (PEG), while fish exposed to PEG did not show any significant difference in aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and acetylcholinesterase (AChE) activities and albumin and globulins levels. Exposure to a combination of chlorpyrifos and PEG caused a significant change in AST, GGT, CPK, LDH and AChE activity, total protein and glucose level. The influence of chlorpyrifos and PEG combination on ALP, cholesterol, triglyceride, albumin, and globulin depends on the concentration of the insecticide and PEG. With a reduction in chlorpyrifos concentration, PEG can maintain ALP activity and albumin at a normal level. Although fish exposure to chlorpyrifos, alone or in combination with polyethylene glycol, significantly increased malondialdehyde (MDA) and catalase (CAT) level, it decreased total antioxidant level (TAN). Exposure to just polyethylene glycol had no impact on CAT activity, TAN and MDA level. According to the results, PEG can have an antagonistic effect on chlorpyrifos toxicity, depending on these two materials concentration. However, chlorpyrifos increased the toxicity of PEG., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Sialic acid is a family of N- and O-substitutions of neuraminic acid. Plasma or serum sialic acid has been established as a potential disease marker. For example, the presence of 9- O-acetyl on the sialic acid of some glycans and glycoconjugates (e.g., 9- O-acetyl GD3 ganglioside) could be related to cancer occurrence. A variety of assays are available to measure serum or plasma sialic acid; however, sample preparation and storage can alter the O-acetylation profile due to the loss of O-acetyl groups and/or the migration of O-acetyl groups. Herein, we report dried blood spot (DBS) sampling, in combination with diamino-4,5-methylenedioxybenzene derivatization, for profiling sialic acids in blood samples with minimal alteration in O-acetylation patterns. The feasibility of the method was first evaluated by analyzing sialic acids in crucian carp blood and comparing with traditional blood/plasma sample preparation procedures. A total of 19 different sialic acids were identified by using liquid chromatography-Orbitrap mass spectrometry, including four mono-O-acetylated N-acetylneuraminic acids, four mono-O-acetylated N-glycolylneuraminic acids, six di-O-acetylated N-acetylneuraminic acids, and three tri-O-acetylated N-acetylneuraminic acids. The long-term storage study indicated that DBS sampling could effectively preserve the O-acetylation information for at least 6 weeks. Thus, it is demonstrated that this method is a valuable tool for the study of sialic acid diversity, especially for the characterization of isomeric structures.

Phenylurea herbicides such as linuron are commonly applied in agriculture. Common carp juveniles were subjected to 31.5 µg/L of linuron for 14 days, and then to 30 days of purification. Peripheral blood was sampled after 1, 3, 7 and 14 days of exposure and 7, 14 and 30 days of purification and hematological parameters were evaluated: erythrocyte (RBCc) and leukocyte (WBCc) counts, hematocrit (Ht), hemoglobin concentration (Hb), mean cell volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) and differential leukocyte count. For evaluation of cortisol and catecholamine concentrations blood was sampled after 3, 6 and 12 h, after 1, 3 and 14 days of exposure, and after 30 days of purification. Linuron caused mainly transient increase in RBCc, Ht and MCV values and increase in WBCc and percentage of juvenile neutrophils. The herbicide caused persistant increase of cortisol and catecholamine concentrations. The results indicate that exposure to low concentration of linuron induced a stress response in common carp.

Fish are the most diverse species of all vertebrate groups, and their blood cells have shown variable characteristics in terms of morphology. Cytochemical staining for enzyme activity in blood leukocytes will help assess the immune function of fish. We characterize blood cells from crucian carp (Carassius auratus) and grass carp (Ctenopharyngodon idellus) by using a Diff-Quick stain as well as different cytochemical methods. Blood specimens obtained from crucian carp and grass carp were evaluated after cytochemical staining for acid phosphatase (ACP), alkaline phosphatase (ALP), naphthol AS chloroacetate esterase (AS-DNCE), naphthyl acetate esterase (NAE), α-naphthyl butyrate esterase (NBE), peroxidase (MPO) and periodic acid-Schiff's reaction (PAS) using commercial kits. Blood cell types were evaluated based on their morphological characteristics and the presence or absence of specific chromogen. The expression pattern of enzymes was similar between the two Cyprinidae and was also broadly consistent with other fish species. However, there were some interesting differences detected between crucian carp and grass carp, including naphthol AS chloroacetate esterase activity in monocytes, peroxidase activity and location in thrombocytes. The ACP, ALP and MPO expressions of different leukocytes of the two Cyprinidae were evaluated by Image Pro Plus and were analysed for statistical significant differences. This investigation provides basic haematology and enzyme activity analyses for crucian carp and grass carp and serves as an approach to evaluating the immune response of fish., (© 2018 Blackwell Verlag GmbH.)

In this study, the effects of some plant hydrosols (distilled plant waters) based upon some hematological parameters and Nitroblue Tetrazolium (NBT) activities in the common carp (Cyprinus carpio Linnaeus, 1758) infected with Yersinia ruckeri were investigated. In the trial, it was utilized totally 200 common carps with 54.3±6.7 g mean live weight and 15.7±1.8 cm mean total lenght. The 10% rate of the common yarrow (Achillea millefolium Linnaeus) hydrosol; 0.5% rate of the cinnamon (Cinnamomum zeylanicum Blume) hydrosol; and 5% rate of the rosemary (Rosemarinus officinalis Linnaeus) hydrosol were applied to fish as a bath treatment. The erythrocyte (RBC), leukocyte count (WBC), hematocrit value (HCT), haemoglobin amount (Hg), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), and activities of NBT in the blood samples taken from the caudal vena of the control and experimental fish groups were analyzed in the 7th, 14th, and 21st days of the exposure treatment. At the end of the research, HCT, Hg, RBC, WBC, MCH and MCV values decreased in the C-2 Group (the control group contain pathogen) compared to the C-1 Group (the control group no contain pathogen), except MCHC value. The NBT activities in the C-1 Groups increased at the 14th day, but decreased quite a few at the 21st day. It has been consequently reached the conclusion that the bath treatments of the some plant hydrosols might be beneficial in increasing of antibacterial properties and in strengthening of defense mechanisms of common carp against Y. ruckeri pathogen.

During the present research, C. idella and C. auratus fish were exposed to 2 ppm concentration of imidacloprid for 28 and 24 days, respectively, and the effect on biochemical and haematological parameters was investigated. During the study of biochemical parameters, there occurred significant increase (P<0.05) in the serum levels of ALT and creatinine of imidacloprid exposed groups of both species of fish. The level of serum albumin of imidacloprid exposed groups of both fish species was significantly lower as compared to control group (P<0.05). Serum globulin level in imidacloprid exposed group of C. idella was insignificantly lower as compared to control group, however the serum globulin level of C. auratus was significantly lower than the control group (P < 0.05). The level of total proteins in serum of imidacloprid exposed groups of both fish species was insignificantly lower as compared to control groups (P>0.05). During the study of haematological parameters, TLC of C. idella was insignificantly (P>0.05) higher than control group but the TLC of C. auratus was significantly (P<0.05) higher than control. There was also observed increasing trend in the percentage of neutrophils and lymphocytes of imidacloprid exposed group of each fish species. The platelets count of imidacloprid exposed group of each fish species was significantly (P<0.05) lower than control group. The haemoglobin concentration of imidacloprid exposed group of C. idella was significantly lower than control group (P<0.05). In case of C. auratus, the haemoglobin level of imidacloprid exposed group was insignificantly lower than control group (P>0.05). From the finding of the present research it was concluded that 28 days exposure of C. idella and 24 days exposure of C. auratus to 2 ppm concentration of imidacloprid does not cause mortality however the exposure causes alteration in the normal level of biochemical and haematological parameters.

Aeromonas veronii is a type of human-livestock-aquatic animal pathogen; it is widely found in nature and causes many deaths among aquatic animals. Extracellular products (ECPs) are secreted by the pathogen during growth and reproduction. These products are considered effective protective antigens that can induce the host to produce an immune response. In this study, the ECPs of A.veronii TH0426 were prepared by ultrafiltration, and then the pathogenicity and enzymatic activity of the ECPs were determined. All the groups were injected intraperitoneally, as follows: group one: ECP protein with an equal volume of Freund's adjuvant; group two: ECPs and formalin-killed cells (FKC) of A.veronii combined with an equal volume of Freund's adjuvant (FKC + ECPs); group three: formalin-killed cells (FKC) of A.veronii combined with an equal volume of Freund's adjuvant (FKC); and, group four: sterile PBS as the control group. The expression levels of IgM, IL-1β, and TNF-α and the lysozyme activity in blood were examined at 7, 14, and 21 days after the immunizations. The results show that the ECPs can produce protease, lipase, amylase and hemolyase, and there was no lecithinase, urease, or gelatinase activity. The results indicate that the ECPs were clearly pathogenic to koi fish, and the LD 50 dose was 391.6 μg/fish. Throughout this study, the RPS of the three experimental groups were 75%, 50%, and 70%. This study indicates that the ECPs of A.veronii can effectively enhance the ability of kio fish to resist bacterial invasion., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Crowding stress is one of the most common environmental stressors in intensive aquaculture. To investigate the influences of long-term crowding stress on nonspecific immune responses and apoptosis in fish, grass carp (Ctenopharyngodon idella) were cultured at low (0.9 kg m -2 ), medium (2.97 kg m -2 ) and high (5.9 kg m -2 ) stocking densities for 10 weeks in the present study. The results showed that elevation of stocking densities caused splenic tissue damages and inflammatory responses, which are characterized with the formation of melano-macrophage centers and the increase of granulocytes as well as significant upregulation of inflammatory cytokine genes (il1β and tnfα). The remarkable decline in the activities of serum lysozyme, acid phosphatase and alkaline phosphatase under high stocking density further confirmed that increased stocking density affected fish nonspecific immune response negatively. Moreover, the transcriptional levels of splenic apoptotic-related genes caspase-8, fasl and caspase-3 increased significantly while the mRNA levels of bax, bcl2, apaf1 and caspase-9 remained unchanged. This result showed that increased stocking density caused splenic cell apoptosis, which were closely associated with the FasL signaling pathway. Our findings revealed that crowding stress could influence fish nonspecific immune response negatively and increase inappropriate apoptosis of the spleen, which would make fish more susceptible to pathogens and ultimately impair fish survival. The breeding density utilized in this study also provides some reference values in intensive aquaculture systems from the perspective of fish health and welfare., (Copyright © 2018. Published by Elsevier Ltd.)

The aim of present study was to evaluate the effects of edible eryngii mushroom powder, Pleurotus eryngii (PE), for 63 days on haematological parameters, the serum immune responses, skin mucus, bactericidal activity, stress resistance, growth performance and digestive enzyme activities of Koi carp fingerlings (Cyprinus carpio koi). Fish were divided into five groups and each group was fed with dietary PE with five graded levels (0, 0.5, 1, 1.5 and 2%). The results showed a significant dose-dependent increase of Ht, Hb, MCV and MCH levels in fish fed dietary PE (P < 0.05). The highest levels of WBCs, lymphocytes and monocytes were measured in fish fed 1.5% and 2% of dietary PE (P < 0.05). The activities of total IG, lysozyme, Alternative haemolytic complement activity in serum of fish fed with 2% of dietary PE for 63 days as well as 5-min air exposure challenge test were significantly higher than other groups (P < 0.05). The most bactericidal activity was observed in skin mucus of fish fed with 1.5% of dietary PE against Streptococcus iniae (P < 0.05). The highest ratio of the lactobacillus count to the total viable count was observed in fish fed 2% of dietary PE. The α-amylase activity of fish fed with dietary PE (1, 1.5 and 2%) were significantly higher than control group (P < 0.05). Feeding fish 2% of dietary PE increased the trypsin and lipase activity compared to others groups (P < 0.05). The growth performance of fish fed 1.5% of dietary PE improved compared to control group (P < 0.05). The results revealed that feeding koi fish with dietary supplementation of PE (1.5 and 2%) improved the selected humoral innate immune responses, bactericidal activity of skin mucus and growth performance of koi fish., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The short term (96 h) toxicity of N-acetyl-p-aminophenol (0.58 mg/L - Treatment I and 0.29 mg/L - Treatment II) on certain health indicators (haematology, biochemical, and enzymology) of an Indian major carp Labeo rohita was studied. When compared to control, N-acetyl-p-aminophenol treated fish showed a significant (P < 0.05) decrease in haemoglobin (Hb), haematocrit (Hct), and erythrocyte (RBC) levels throughout the study period. Whereas, a significant (P < 0.05) increase were noted in leucocyte (WBC) counts (except 48 h in Treatment-I), mean corpuscular volume (MCV), and mean corpuscular haemoglobin (MCH) values (except 24 h in Treatment-I). Mean corpuscular haemoglobin concentration (MCHC) values were found to be decreased significantly (P < 0.05) in fish exposed to 0.58 mg/L of N-acetyl-p-aminophenol, whereas in 0.29 mg/L exposed fish the values were found to be increased significantly (P < 0.05) (except 72 h). A significant (P < 0.05) increase in plasma glucose levels was noticed in fish exposed to 0.58 mg/L of N-acetyl-p-aminophenol (except 96 h). However, a biphasic trend in plasma glucose level was observed at 0.29 mg/L of N-acetyl-p-aminophenol exposed fish. Protein levels were found to be increased in both the treatments except at the end of 48 and 96 h in 0.58 and 0.29 mg/L, respectively. In both the treatments fluctuations of enzyme (GOT, GPT, and LDH) activities in gill and liver were also noted. However, these enzyme activities found to be significantly (P < 0.05) decreased in kidney and plasma of fish. From the result we conclude that the drug N-acetyl-p-aminophenol upon short term exposure could pose a risk to fish and the alteration of these parameters can be used to ecological risk assessment of pharmaceuticals in aquatic organisms., (Copyright © 2018 Elsevier Inc. All rights reserved.)

An 8-week feeding trial was conducted to compare the effects of supplementing (0.7 mg kg -1 ) different dietary selenium (Se) sources including organic [selenomethionine (SeMet)], inorganic [sodium selenite (Na 2 SeO 3 )], and nanoparticulate Se (nano-Se) on physiological responses of common carp, Cyprinus carpio juveniles (9.7 ± 0.1 g). Basal diet without Se supplementation used as control. Fish fed nano-Se supplemented diet had the highest weight gain (97.2 ± 10.8%) and feed efficiency ratio (42.4 ± 0.8%). Intestinal villi height was significantly taller in fish fed nano-Se diet than in the control group in both foregut and midgut sections. Serum glutathione peroxidase and superoxide dismutase activities were significantly higher in nano-Se and SeMet groups than in control and sodium selenite groups. Fish fed Se-supplemented diets had greater red blood cell counts and hematocrit and hemoglobin values than the control group (P < 0.05). Nano-Se and SeMet groups showed a significant increase in white blood cell counts, neutrophil percentage, and serum lysozyme activity than the other groups. Fish fed nano-Se diet had the highest serum hemolytic activity, total immunoglobulin, and total protein and albumin contents, as well as the lowest serum total cholesterol and low density lipoprotein levels (P < 0.05). Overall, significant improvements in growth performance, feed utilization, intestinal morphology, and hemato-immunological and serum biochemical parameters of common carp juveniles suggest nano-Se as an efficient source for providing dietary Se in this species.

Trichlorfon is a moderately toxic organophosphate pesticide that is widely used in aquaculture. This study investigated the effects of trichlorfon on hematological parameters, biochemical factors, and stress reaction in Cyprinus carpio L. The fish were exposed to acute concentrations of trichlorfon (0, 0.5, 1.0, 2.0, and 4.0 mg L -1 ) at 25 °C and 15 °C for 1 and 2 weeks, after which several parameters were evaluated to assess the effects of the pesticide. Significant decreases were observed in red blood cell (RBC) Count, hemoglobin (Hb) level, hematocrit (Ht), and plasma protein levels after each exposure period. In contrast, notable increases in mean corpuscular volume (MCV), mean cell hemoglobin (MCH), calcium, and glucose levels were observed in the trichlorfon-treated groups. Additionally, there were significant increases in the plasma levels of glutamate-oxaloacetate transaminase (GOT), glutamate-pyruvate transaminase (GPT), and alkaline phosphatase (ALP) following the exposure to trichlorfon. Furthermore, the results showed a relationship between toxic stress and increment in HSP70 and cytochrome P450 1A (CYP1A) expression over time. Ht, MCV, MCH, and the value of other biochemical parameters were quite lower at 15 °C than their corresponding values were at 25 °C, which indicated the decreased physical activity at 15 °C. The results of the present work indicate that acute exposure to trichlorfon and thermal stimulus could damage erythropoietic tissue. Additionally, hepatocytes function and physiological mechanisms could be impaired. Ht, glucose, GOT, GPT, HSP70, and CYP1A levels might be useful biomarkers of trichlorfon toxicity in contaminated aquatic ecosystems., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The effects of Roundup (glyphosate-based herbicide) on peripheral blood parameters and hematopoietic tissue cellular composition and activity in common carp juveniles were evaluated. The fish were exposed for 7 days at 0.0, 0.1, 0.5, or 5.0 mg/L of glyphosate. In fish exposed to Roundup, hematological alterations were observed; however, most of them were not directly related to the herbicide concentration. An increase in Ht and MCV, and decrease in Hb, MCH, and MCHC compared to the control were observed. Fish exposed to Roundup showed also a reduction in WBC and oxidative metabolic activity of phagocytes (NBT) compared to the control. The fish exposed to 0.1 and 5.0 mg/L showed increased glucose values, whereas in those subjected to 0.5 mg/L blood glucose concentration declined compared to the control. Cholesterol significantly increased at 0.1 mg/L and decreased at 5.0 mg/L. Analysis of head kidney hematopoietic tissue revealed that Roundup at concentrations 0.5 and 5.0 mg/L caused a significant increase in the rate of cell proliferation accompanied by an increase in frequency of early blast cells. No significant differences occurred in percentages of most cell lineages but the frequency of monocytoid, eosinophilic, and basophilic lineage cells significantly increased in the herbicide-exposed fish compared to the control. The obtained results revealed that sublethal concentrations of Roundup that may occur in polluted natural waters caused a slight anemic and significant immunosuppressive response in common carp juveniles. On the other hand, they indicate a considerable compensatory potential of carp hematopoietic system.

The environmental temperature affects plasma biochemical indicators, antioxidant status and hematological and immunological parameters in fish. So far, only single blood proteins have been identified in response to temperature changes. The aim of this study was to compare the proteome of carp blood plasma from males acclimated to warm (30 °C) and cold (10 °C) temperatures by two-dimensional differential gel electrophoresis followed by MALDI-TOF/TOF mass spectrometry. A total of 47 spots were found to be differentially regulated by temperature (>1.2-fold change, p < 0.05): 25 protein spots were more abundant in warm-acclimated males and 22 were enriched in cold-acclimated males. The majority of differentially regulated proteins were associated with acute phase response signalling involved in: i) activation of the complement system (complement C3-H1), ii) neutralization of proteolytic enzymes (inter-alpha inhibitor H3, fetuin, serpinA1, antithrombin, alpha2-macroglobulin), iii) scavenging of free hemoglobin and radicals (haptoglobin, Wap65 kDa), iv) clot-formation (fibrinogen beta and alpha chain, T-kininogen) and v) the host's immune response modulation (ApoA1 and ApoA2). However, quite different sets of these proteins or proteoforms were involved in response to cold and warm temperatures. In addition, cold acclimation seems to be related to the proteins involved in lipid metabolism (apolipoproteins A and 14 kDa) and stress response (corticosteroid binding globulin). We discovered a strongly regulated protein Cap31 upon cold acclimation, which can serve as a potential blood biomarker of cold response in carp. These studies significantly extend our knowledge concerning mechanisms underlying thermal adaptation in poikilotherms., (Copyright © 2018. Published by Elsevier Ltd.)

The aim of this study was to determine protective effects of curcumin on some haematological values and oxidant/antioxidant status in Cyprinus carpio exposed to chlorpyrifos. The fish were exposed to two sublethal concentrations of chlorpyrifos (0.040 and 0.080mgL), and curcumin (100mg per kg of fish weight) was simultaneously administered for 14 days. Blood and tissue (liver, kidney, and gill) samples were collected at the end of the experiment and analysed to determine the haematological profile (red blood cell count, white blood cell count, haemoglobin concentration, and haematocrit level) and oxidant/antioxidant status (malondialdehyde level and superoxide dismutase, catalase, glutathione peroxidase, and glutathione-S-transferase activities) of the fish. There was a significant decrease in the red blood cell count, the haemoglobin concentration, and the haematocrit level and a increase in the white blood cell count of CPF-treated fish. The results revealed a significant increase in the malondialdehyde levels of the groups that were exposed to CPF. Conversely, the MDA levels were significantly decreased by curcumin. Also, CPF exposure caused a significant increase in the superoxide dismutase and glutathione-S-transferase activities and a significant decrease in the catalase and glutathione peroxidase activities. However, curcumin reversed the superoxide dismutase, catalase, glutathione peroxidase, and glutathione-S-transferase activities. In conclusion, this study demonstrated that CPF had a negative effect on the haematological values and the oxidant/antioxidant status of the fish. The simultaneous administration of curcumin was neutralised CPF-induced toxicity., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The negative impacts of residual antibiotics in the environment on ecosystem and human health are big concerns. However, little information is available on the antibiotic bioaccumulation in aquaculture farms. In this study, the bioaccumulative potentials of 21 antibiotics in the plasma, bile, liver and muscle of cultured fish from a typical aquaculture area were systematically investigated., Results: indicated that antibiotic distribution in the cultured fish was mainly influenced by species and specific substances. The mean values of log bioaccumulation factors (Log BAFs) for the detected antibiotics were in the range of 0.43-3.70, 0.36-4.75, -0.31-4.48, and 0.23-4.33 in the fish plasma, bile, liver and muscle tissues, respectively. For grass carp, both ciprofloxacin and enrofloxacin showed high transportability from the plasma to the muscle and liver. The correlations of various antibiotic concentrations between the plasma and the other three tissues indicated that the concentrations of ciprofloxacin and enrofloxacin in the fish tissues could be predicted by their concentrations in the plasma. Based on the calculated hazard quotients, human health risk evaluation of antibiotic exposure by fish consumption indicated that the consumption of these cultured fish posed low risks to human health., (Copyright © 2017 Elsevier B.V. All rights reserved.)