A study on assessing the toxic effects of ethyl paraben on rohu (Labeo rohita) using different biomarkers; hemato-biochemical assays, histology, oxidant and antioxidant activity and genotoxicity.

Parabens are being used as preservatives due to their antifungal and antimicrobial effects. They are emerging as aquatic pollutants due to their excessive use in many products. The purpose of this study was to determine the toxic effect of ethyl paraben (C9H10O3) on the hematobiochemical, histological, oxidative, and anti-oxidant enzymatic and non-enzymatic activity; the study also evaluates the potential of ethyl paraben to cause genotoxicity in Rohu Labeo rohita. A number of 15 fish with an average weight of 35.45±1.34g were placed in each group and exposed to ethyl paraben for 21 days. Three different concentrations of ethyl paraben, i.e., T1 (2000µg/L), T2 (4000 µg/L), andT3 (6000 µg/L) on which fish were exposed as compared to the control T0 (0.00 µg/L). Blood was used for hematobiochemical and comet assay. Gills, kidneys, and liver were removed for histological alterations. The results showed a significant rise in all hemato-biochemical parameters such as RBCs, WBCs, PLT count, blood sugar, albumin, globulin, and cholesterol. An increase in aspartate aminotransferase (AST) and alanine transaminase (ALT) levels directed the hepatocytic damage. Histological alterations in the liver, gills and kidneys of fish were found. Ethylparaben induces oxidative stress by suppressing antioxidant enzyme activity such as SOD, GSH, CAT and POD. Based on the comet assay, DNA damage was also observed in blood cells, resulting in genotoxicity. Findings from the present study indicate that ethyl paraben induces hemato-biochemical alterations, tissue damage, oxidative stress, and genotoxicity.

Evaluation of the toxicological properties of Himantura imbricata Venom using a zebrafish model (Danio rerio).

The stingrays of the genus Himantura imbricata are present in all of the world's oceans, but the toxicity of their venoms has not yet been thoroughly characterized. The zebrafish as a toxicology model can be used for general toxicity testing of drugs and the investigation of toxicological mechanisms. The aim of this study was to evaluate the effect of crude venom from the stingray H. imbricata on the zebrafish Danio rerio. Juvenile zebrafish were injected with different concentrations of venom from H. imbricata via subcutaneous injections. The venom's effects were established via histological examination and hemolytic activity in zebrafish. The histopathological analysis revealed significant tissue damage in the organs of the zebrafish injected with venom, including liver necrosis and kidney degeneration. A blood examination revealed echinocytes, hemolysis, and nuclear abnormalities. Bodyweight estimations and histopathological attributes of the gills, heart, muscle, liver, intestine, eye, and brain were determined. The histological staining studies of the gills, liver, and intestine were measurably higher in the venom groups compared with the other two groups. Aggregately, the result shows that zebrafish may act as a valuable biomarker for alterations impelled by H. imbricata venom. The work delivers a useful model with substantial pharmacological potential for new drugs and a better comprehension of research on stingray venom.

Mucosal barrier status in Atlantic salmon fed rapeseed oil and Schizochytrium oil partly or fully replacing fish oil through winter depression.

The study was designed to investigate the effects of replacing fish oil by algal oil and rapeseed oil on histomorphology indices of the intestine, skin and gill, mucosal barrier status and immune-related genes of mucin and antimicrobial peptide (AMP) genes in Atlantic salmon (Salmo salar). For these purposes, Atlantic salmon smolts were fed three different diets. The first was a control diet containing fish oil but no Schizochytrium oil. In the second diet, almost 50 % of the fish oil was replaced with algal oil, and in the third diet, fish oil was replaced entirely with algal oil. The algal oil contained mostly docosahexaenoic acid (DHA) and some eicosapentaenoic acid (EPA). The study lasted for 49 days in freshwater (FW), after which some fish from each diet group were transferred to seawater (SW) for a 48-h challenge test at 33 ppt to test their ability to tolerate high salinity. Samples of skin, gills, and mid intestine [both distal (DI) and anterior (AI) portions of the mid intestine] were collected after the feeding trial in FW and after the SW-challenge test to assess the effects of the diets on the structure and immune functions of the mucosal surfaces. The results showed that the 50 % VMO (Veramaris® algal oil) dietary group had improved intestinal, skin, and gill structures. Principal component analysis (PCA) of the histomorphological parameters demonstrated a significant effect of the algal oil on the intestine, skin, and gills. In particular, the mucosal barrier function of the intestine, skin, and gills was enhanced in the VMO 50 % dietary group after the SW challenge, as evidenced by increased mucous cell density. Immunolabelling of heat shock protein 70 (HSP70) in the intestine (both DI and AI) revealed downregulation of the protein expression in the 50 % VMO group and a corresponding upregulation in the 100 % VMO group compared to 0 % VMO. The reactivity of HSP70 in the epithelial cells was higher after the SW challenge compared to the FW phase. Immune-related genes related to mucosal defense, such as mucin genes [muc2, muc5ac1 (DI), muc5ac1 (AI), muc5ac2, muc5b (skin), and muc5ac1 (gills)], and antimicrobial peptide genes [def3 (DI), def3 (AI), and cath1 (skin)] were significantly upregulated in the 50 % VMO group. PCA of gene expression demonstrated the positive influences on gene regulation in the 50 % VMO dietary group. In conclusion, this study demonstrated the positive effect of substituting 50 % of fish oil with algal oil in the diets of Atlantic salmon. The findings of histomorphometry, mucosal mapping, immunohistochemistry, and immune-related genes connected to mucosal responses all support this conclusion.

Untargeted LC-MS metabolomics reveals the metabolic responses in the Eriocheir sinensis gills exposed to salinity and alkalinity stress.

In recent years, saline-alkaline aquaculture development has become an important measure for China to expand its fishery development space to ensure food safety. Previous studies have verified that salinity and alkalinity positively influence the quality of Chinese mitten crabs (Eriocheir sinensis). However, the regulatory mechanism of E. sinensis endures saline-alkaline stress which remains obscure. This study investigated the metabolic changes in puberty-molting E. sinensis gills exposed to freshwater (FW), sodium chloride salinity of 5 ppt (SW), and carbonate alkalinity 10.00 mmol/L (AW) for 50 days using untargeted liquid chromatography-mass spectrometry metabolomics (LC-MS). A total of 5802 (positive-ion mode) and 6520 (negative-ion mode) peaks were extracted by LC-MS, respectively. A total of 188 (50 upregulated and 138 downregulated), 141 (94 upregulated and 47 downregulated), and 130 (87 upregulated and 43 downregulated) significantly regulated metabolites (SRMs) were observed in the FW-SW, FW-AW, and SW-AW treatments, respectively, wherein 42 generic SRMs were also found by Venn diagram analysis. Seven of the top 10 SRMs with the highest (variable importance in projection) VIP values were similarly identified in FW-SW and SW-AW. Integrated analysis of key metabolic pathways revealed glycerophospholipid, choline in cancer, phenylalanine, and butanoate metabolism. Overall, significant differences were observed in the metabolites and key metabolic pathways of E. sinensis gill exposed to salinity and alkalinity stress. These results will be helpful in understanding the environmental adaptability of aquatic crustaceans to saline-alkaline water.

Toxic effects of aniline in liver, gills and kidney of freshwater fish Channa punctatus after acute exposure.

Aniline (C6H5NH2) is one of the hazardous aromatic amine where an amino group -NH2) is connected to phenyl ring (C6H5). Based on the evaluation of the 96-hour LC50 of aniline, two sublethal concentrations (4.19 mg/l and 8.39 mg/l) were selected for acute exposure tests in freshwater fish Channa punctatus. The liver, gills and kidney of fish being the principal sites of xenobiotic material accumulation, respiration, biotransformation, and excretion are the focus of the present study. Throughout the exposure time, the comet assay revealed increased tail length and tail DNA percentage indicating maximum damage to liver, gills and kidney of treated group after 96 h. After acute exposure, there was a significant (p ≤ 0.05) increase in the enzymatic activity of glutathione-S-transferase (GST) and acetylcholinesterase (AChE), whereas decline in superoxide dismutase (SOD) and catalase (CAT) activity was observed. Meanwhile, levels of malondialdehyde (MDA) increased over the exposure period for both concentrations. After 96 h of exposure, degree of tissue change (DTC) was evaluated in liver, gill and kidney of aniline exposed fish. Additionally, light microscopy revealed multiple abnormalities in liver, gills and kidney of all the treated groups. Significant changes were observed in the levels of biochemical markers viz., glucose, triglyceride, cholesterol, aspartate transaminase, alanine transaminase and urea following a 96-hour exposure to aniline. Studies using ATR-FTIR and transmission electron microscopy (TEM) revealed changes in biomolecules and structural abnormalities in several tissues of the aniline-exposed groups in comparison to the control group respectively.

Biological toxicity of sulfamethoxazole in aquatic ecosystem on adult zebrafish (Danio rerio).

This study evaluated the impacts of sulfamethoxazole (SMX) on antioxidant, immune, histopathological dynamic changes, and gut microbiota of zebrafish. SMX was carried out five groups: 0 (C), 3 mg/L (T3), 6 mg/L (T6), 12 mg/L (T12), and 24 mg/L (T24), with 5 replicates per group for an 8-weeks chronic toxicity test. It was found that SMX is considered to have low toxicity to adult zebrafish. SMX with the concentration not higher than 24 mg/L has no obvious inhibitory effect on the growth of fish. Under different concentrations of SMX stress, oxidative damage and immune system disorder were caused to the liver and gill, with the 12 and 24 mg/L concentration being the most significant. At the same time, it also causes varying degrees of pathological changes in both intestinal and liver tissues. As the concentration of SMX increases, the composition and abundance of the gut microbiota in zebrafish significantly decrease.

A comparative study on targeted gene expression in zebrafish and its gill cell line exposed to chlorpyrifos.

Chlorpyrifos (CPF) is an organophosphorus-based insecticide, which is known to pose a serious risk to aquatic animals. However, the mechanisms of CPF toxicity in animals still remain unclear. The present investigation aimed to compare the potential effects of CPF in zebrafish (Danio rerio) and its gill cell line (DrG cells). Based on the in vivo study, the LC50 was calculated as 18.03 µg/L and the chronic toxic effect of CPF was studied by exposing the fish to 1/10th (1.8 µg/L) and 1/5th (3.6 µg/L) of the LC50 value. Morphological changes were observed in fish and DrG cells which were exposed to sublethal concentrations of CPF. The results of MTT and NR assays showed significant decline in the survival of cells exposed to CPF at 96 h. The production of reactive oxygen species in DrG cells and expression levels of antioxidant markers, inflammatory response genes (cox2a and cox2b), cyp1a, proapoptotic genes (bax), antiapoptotic gene (bcl2), apoptotic genes (cas3 and p53), and neuroprotective gene (ache) were determined in vivo using zebrafish and in vitro using DrG cells after exposure to CPF. Significant changes were found in the ROS production (DrG cells) and in the expression of inflammatory, proapoptotic, and apoptotic genes. This study showed that DrG cells are potential alternative tools to replace the use of whole fish for toxicological studies.

Assessing the negative impact of chlorantraniliprole, isoxaflutole, and simazine pesticides on phospholipid membrane models and tilapia gill tissues.

The indiscriminate and, very often, incorrect use of pesticides in Brazil, as well as in other countries, results in severe levels of environmental pollution and intoxication of human life. Herein, we studied plasma membrane models (monolayer and bilayer) of the phospholipid Dioleoyl-sn-glycerol-3-phosphocholine (DOPC) using Langmuir films, and large (LUVs) and giant (GUVs) unilamellar vesicles, to determine the effect of the pesticides chlorantraniliprole (CLTP), isoxaflutole (ISF), and simazine (SMZ), used in sugarcane. CLTP affects the lipid organization of the bioinspired models of DOPC π-A isotherms, while ISF and SMZ pesticides significantly affect the LUVs and GUVs. Furthermore, the in vivo study of the gill tissue in fish in the presence of pesticides (2.0 × 10-10 mol/L for CLTP, 8.3 × 10-9 mol/L for ISF, and SMZ at 9.9 × 10-9 mol/L) was performed using optical and fluorescence images. This investigation was motivated by the gill lipid membranes, which are vital for regulating transporter activity through transmembrane proteins, crucial for maintaining ionic balance in fish gills. In this way, the presence of phospholipids in gills offers a model for understanding their effects on fish health. Histological results show that exposure to CLTP, ISF, and SMZ may interfere with vital gill functions, leading to respiratory disorders and osmoregulation dysfunction. The results indicate that exposure to pesticides caused severe morphological alterations in fish, which could be correlated with their impact on the bioinspired membrane models. Moreover, the effect does not depend on the exposure period (24h and 96h), showing that animals exposed to pesticides for a short period suffer irreparable damage to gill tissue. In summary, we can conclude that the harm caused by pesticides, both in membrane models and in fish gills, occurs due to contamination of the aquatic system with pesticides. Therefore, water quality is vital for the preservation of ecosystems.

Nano-titanium dioxide exacerbates the harmful effects of perfluorooctanoic acid on the health of mussels.

Exposing marine organisms to contemporary contaminants, such as perfluorooctanoic acid (PFOA) and nano-titanium dioxide (nano-TiO2), can induce multifaceted physiological consequences. Our investigation centered on the responses of the mussel, Mytilus coruscus, to these agents. We discerned pronounced disruptions in gill filament connections, pivotal structures for aquatic respiration, suggesting compromised oxygen uptake capabilities. Concurrently, the respiratory rate exhibited a marked decline, indicating a respiratory distress. Furthermore, the mussels' clearance rate, a metric of their filtration efficacy, diminished, suggesting the potential for bioaccumulation of deleterious substances. Notably, the co-exposure of PFOA and nano-TiO2 exhibits interactive effects on the physiological performance of the mussels. The mussels' digestive performance waned in the face of heightened PFOA and nano-TiO2 concentrations, possibly hampering nutrient assimilation and energy accrual. This was mirrored in the noticeable contraction of their energy budget, suggesting long-term growth repercussions. Additionally, the dysregulation of the gut microbiota and the reduction in its diversity further confirm alterations in intestinal homeostasis, subsequently impacting its physiological functions and health. Collectively, these findings underscore the perils posed by escalated PFOA and nano-TiO2 levels to marine mussels, accentuating the need for a deeper understanding of nanoparticle-pollutant synergies in marine ecosystems.

A realistic combined exposure scenario: effect of microplastics and atrazine on Piaractus mesopotamicus.

Microplastics, considered emerging environmental contaminants resulting from plastic degradation, are discovered in diverse aquatic ecosystems and can be unintentionally ingested by fish. Therefore, it is essential to characterize their interaction with other contaminants, such as agrochemicals, in aquatic environments. This study aimed to assess histological, enzymatic, and genotoxic biomarkers in juvenile pacú (Piaractus mesopotamicus) exposed to polyethylene (PE) microplastic particles and the herbicide atrazine, individually or combined, for 15 days. Four treatments were used: a negative control (CON), PE in the fish diet (0.1% w/w, FPE), atrazine through water (100 µg L-1, ATZ), and the mixture (ATZ+FPE). Results confirmed histological alterations in gills (edema and lamellar fusion) and liver (necrotic areas and congestion) of fish exposed to ATZ and ATZ+FPE. The number of goblet cells increased in the posterior intestine of fish under ATZ+FPE compared to CON and FPE. Enzyme activities (CAT, GST, AChE, and BChE) significantly increased in ATZ+FPE compared to CON. However, no genotoxic effect was demonstrated. These findings provide insights into the complex impacts of simultaneous exposure to atrazine and microplastics, emphasizing the need for continued research to guide effective environmental management strategies against these contaminants that represent a risk to aquatic organisms.

Assessment of the water quality and toxicity effects on zebrafish (Danio rerio) of a stream near a phosphorus chemical plant in Guizhou Province, southwestern China.

To reveal the influence of the phosphorus chemical industry (PCI) on regional water environmental quality and safety, the water quality and ecotoxicological effects of a stream near a phosphorus chemical plant (PCP) in Guizhou Province, southwestern China, were investigated based on water samples collected from the stream. The results showed that the average concentrations of NH3-N, TN, P, F-, Hg, Mn, and Ni were 3.14 mg/L, 30.09 mg/L, 3.34 mg/L, 1.18 mg/L, 1.06 µg/L, 45.82 µg/L, and 11.30 µg/L, respectively. The overall water quality of the stream was in the heavily polluted category, and NH3-N, TN, P, F-, and Hg were the main pollution factors. The degree of pollution was in the order of rainy period > transitional period > dry period, and the most polluted sample site was 1100 m from the PCP. After 28 days of exposure to stream water, there was no significant change in the growth parameters of zebrafish. The gills of zebrafish showed a small amount of epithelial cell detachment and a small amount of inflammatory cell infiltration, and the liver tissue displayed a large amount of hepatocyte degeneration with loose and lightly stained cytoplasm. Compared with the control group, the %DNA in tail, tail length, tail moment, and olive tail moment were significantly increased (p < 0.05), indicating that the water sample caused DNA damage in the peripheral blood erythrocytes of zebrafish. The stream water in the PCI area was found to be polluted and exhibited significant toxicity to zebrafish, which could pose a threat to regional ecological security.

Bisphenol A Induces Histopathological, Hematobiochemical Alterations, Oxidative Stress, and Genotoxicity in Common Carp (Cyprinus carpio L.).

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.

Effects of Cadmium Exposure on Gut Villi in Danio rerio.

In aquatic organisms, cadmium exposure occurs from ovum to death and the route of absorption is particularly wide, being represented by skin, gills and gastrointestinal tract, through which contaminated water and/or preys are ingested. It is known that cadmium interferes with the gut; however, less information is available on cadmium effects on an important component of the gut, namely goblet cells, specialized in mucus synthesis. In the present work, we studied the effects of two sublethal cadmium concentrations on the gut mucosa of Danio rerio. Particular attention was paid to changes in the distribution of glycan residues, and in metallothionein expression in intestinal cells. The results show that cadmium interferes with gut mucosa and goblet cells features. The effects are dose- and site-dependent, the anterior gut being more markedly affected than the midgut. Cadmium modifies the presence and/or distribution of glycans in the brush border and cytoplasm of enterocytes and in the goblet cells' cytoplasm and alters the metallothionein expression and localization. The results suggest a significant interference of cadmium with mucosal efficiency, representing a health risk for the organism in direct contact with contamination and indirectly for the trophic chain.

Oxidative stress responses after exposure to triclosan sublethal concentrations: an integrated biomarker approach with a native (Corydoras paleatus) and a model fish species (Danio rerio).

Triclosan (TCS) is a synthetic broad-spectrum antimicrobial agent commonly used world-wide in a range of personal care and sanitizing products detected frequently in aquatic ecosystems. The aim of this study was to examine biochemical markers responses triggered by TCS in Danio rerio and in a native South American fish species (Corydoras paleatus). Further, an integrated approach comparing both test fish species was undertaken. These fish organisms were exposed to 100 or 189 µg TCS/L for 48 h. The activities of catalase (CAT), glutathione-s-transferase (GST), superoxide dismutase (SOD), and lipid peroxidation levels (LPO) and total antioxidant capacity against peroxyl radicals (ACAP) were determined in liver, gills, and brain. Acetylcholinesterase activity (AChE) was measured in the brain. Multivariate analysis showed that the most sensitive hepatic parameters were activities of GST and SOD for C. paleatus while LPO levels were for D. rerio. In gills the same parameters were responsive for C. paleatus but CAT in D. rerio. ACAP and GST activity were responsive parameters in brain of both species. Integrated biomarker responses (IBR) index demonstrated similar trends in both species suggesting this parameter might serve as a useful tool for quantification of integrated responses induced by TCS.

Evaluation of cardiotoxicity in Amazonian fish Bryconops caudomaculatus by acute exposure to aluminium in an acidic environment.

Aluminium (Al) is soluble in acidic waters and may become toxic to organisms. In this study, the acute effects of two Al concentrations were evaluated in the Amazonian fish Bryconops caudomaculatus. Antioxidant responses and lipid damage were assessed in gills, liver and muscle, along with the electrocardiography (ECG) and characterization of cardiac complex and wave intervals. Fish were essayed as follows: two control groups at neutral and acidic pH and two exposure groups at acidic pH (0.3 mg/L and 3.0 mg/L Al). Water samples were collected at 0h, 24h and 48h, for chloride (Cl-), fluoride (F-) and sulphate (SO42-) ion analyses, while total Al was quantified in muscle. Concentrations of Cl- and SO42- were constant over time whereas F- was not detected. Total Al concentrations in water and muscle were concentration-dependent. Antioxidant responses, total antioxidant capacity against peroxyl radicals (ACAP) and glutathione S-transferase were not triggered in fish tissues exposed to 0.3 mg/L Al; however, fish exposed to 3.0 mg/L Al presented increased and reduced ACAP in gills and liver, respectively. No changes in lipoperoxidation levels occurred among groups. Fish exposed to 0.3 mg/L Al showed prolonged intervals in ECG as a reflection of low heart rate (HR), with sinus bradycardia. Moreover, there was a marked prolongation of the PQ interval (time between the atrial activity and the start of ventricular activity), indicating interference on the cardiac cell automaticity. Fish exposed to the highest concentration of Al showed reduced wave intervals as a consequence of increased HR, with sinus arrhythmia, while ECG tracings did not present P waves (atrial contraction), indicating an atrioventricular blockade. In conclusion, 48h exposure sufficed to cause cardiotoxicity in B. caudomaculatus at either Al concentration. However, as oxidative stress was not observed, such cardiac alterations seem to be reversible under the experimental conditions established herein.

Oxidative Stress and Toxico-Pathic Branchial Lesions in Cyprinus carpio Exposed to Malachite Green.

Gill is the frontier tissue to come in direct contact with aquatic toxicants. Malachite green (MG) commercial textile dye was assessed for its impact on the gill cytoarchitecture. Cyprinus carpio were exposed to 0.087 and 0.146 mg/L of MG for 60 days. The tissue was processed, and HE stained slides revealed histo-pathic lesions such as lamellar curling, edema, necrosis, telangiectasia, aneurysm, and vacuolization. Scanning electron microscopy reported aberrations in lamellae and microridges of the epithelium. At the cellular level, transmission electron microscopy exhibited nuclear alterations in form of pyknosis and mitochondrial swelling followed by cristolysis. Pillar cells displayed cytoplasmic vacuolization and leukocyte infiltration, and goblet cell containing varied shaped and density mucous globules. The biochemical analysis supported the ultrastructural alterations and showed a negative impact of MG on the antioxidative enzymes (CAT, SOD, GSH), while levels of MDA were found to be significantly elevated. Thereby, concluding MG induced branchial toxicity in the fish.

The effect of renin-angiotensin-aldosterone system inhibitors on organ-specific ace2 expression in zebrafish and its implications for COVID-19.

Among cases of SARS-CoV-2 infections that result in serious conditions or death, many have pre-existing conditions such as hypertension and are on renin-angiotensin-aldosterone system (RAAS) inhibitors. The angiotensin-converting-enzyme-2 (ACE2), a key protein of the RAAS pathway, also mediates cellular entry of SARS-CoV-2. RAAS inhibitors might affect the expression levels of ace2, which could impact patient susceptibility to SARS-CoV-2. However, multi-organ-specific information is currently lacking and no species other than rodents have been examined. To address this knowledge gap, we treated adult zebrafish with the RAAS inhibitors aliskiren, olmesartan, and captopril for 7 consecutive days and performed qRT-PCR analysis of major RAAS pathway genes in the brain, gill, heart, intestine, kidney, and liver. Both olmesartan and captopril significantly increased ace2 expression in the heart, gill, and kidney. Olmesartan also increased ace2 expression in the intestine. Conversely, aliskiren significantly decreased ace2 expression in the heart. Discontinuation of compound treatments for 7 days did not return ace2 expression to baseline levels. While potential risks or benefits of antihypertensive RAAS inhibitors to SARS-CoV-2 infections in humans remain uncertain, this study provides new insights regarding the impact of RAAS inhibitors on organ-specific ace2 expression in another vertebrate model, thereby providing comparative data and laying scientific groundwork for future clinical decisions of RAAS inhibitor use in the context of COVID-19.

Metaplasia of respiratory and digestive tissues in the eastern oyster Crassostrea virginica associated with the Deepwater Horizon oil spill.

Metaplasia is a well documented and deleterious effect of crude oil components on oysters. This reversible transformation of one cell type to another is a common response to petroleum-product exposure in molluscs. It has been shown experimentally in previous work that eastern oysters (Crassostrea virginica) exposed to petroleum products will exhibit metaplasia of digestive tissues. Here we document for the first time that wild adult oysters inhabiting coastal waters in the northern Gulf of Mexico during and in the aftermath of the Deepwater Horizon oil spill (2010) exhibited metaplasia in both ctenidial (respiratory and suspension feeding) and digestive tract tissues at significantly higher frequencies than geographic controls of C. virginica from Chesapeake Bay. Metaplasia included the loss of epithelial cilia, transformations of columnar epithelia, hyperplasia and reduction of ctenidial branches, and vacuolization of digestive tissues. Evidence for a reduction of metaplasia following the oil spill (2010-2013) is suggestive but equivocal.

Transcriptomic analyses of the acute aerial and ammonia stress response in the gill and liver of large-scale loach (Paramisgurnus dabryanus).

The large-scale loach (Paramisgurnus dabryanus) is one of the most commercially important cultured species. Ammonia nitrogen accumulation is one of the key issue which limited production and animal health in aquaculture, but few of information is available on the molecular mechanisms of ammonia detoxification. We performed transcriptomic analyses of the gill and liver of large-scale loach subjected to 48 h of aerial and ammonia exposure. We obtained 47,473,424 to 56,791,496 clean reads from the aerial exposure, ammonia exposure and control groups, assembled and clustered a total of 92,658 unigenes with an average length of 909 bp and N50 of 1787 bp. Totals of 489/145 and 424/140 differentially expressed genes (DEGs) were detected in gill/liver of large-scale loach after aerial and ammonia exposure through comparative transcriptome analyses, respectively. In addition, totals of 43 gene ontology (GO) terms and 266 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were identified. After aerial and ammonia exposure, amino acid metabolism pathways in liver of large-scale loach were significantly enriched, suggesting that large-scale loach responded to high exogenous and endogenous ammonia stress by enhancing amino acid metabolism. Besides, the expression of several ammonia transporters (i.e., Rhesus glycoproteins and Aquaporins) in gill of large-scale loach were markedly changed after 48 h of aerial exposure, suggesting that large-scale loach responded to high endogenous ammonia stress by regulating the expression of Rh glycoproteins and Aqps related genes in gill. The results provide valuable information on the molecular mechanism of ammonia detoxification of large-scale loach to endogenous and environmental ammonia loading, will facilitate the molecular assisted breeding of ammonia resistant varieties, and will offer beneficial efforts for establishing an environmental-friendly and sustainable aquaculture industry.

Bioremediation of hemotoxic and oxidative stress induced by polyethylene microplastic in Clarias gariepinus using lycopene, citric acid, and chlorella.

Despite extensive research on the toxic effects of microplastics (MPs), there is no obtainable data on the use of phytobioremediation against MPs toxicity in fish. This study aimed to investigate the protective role of lycopene, citric acid, and chlorella against the toxic effects of MPs in African catfish (Clarias gariepinus) using hematology, biochemical, antioxidants, erythron profiles (poikilocytosis and nuclear abnormalities) and the accumulation of MPs in tissues as biomarkers. Five groups of fish received: normal diet (control); MPs (500 mg/kg diet) (Group 2); MPs (500 mg/kg diet) + lycopene (500 mg/kg diet) (Group 3); MPs (500 mg/kg diet) + citric acid (30 g/kg diet) (Group 4); and MPs (500 mg/kg diet) + chlorella (50 g/kg diet) (Group 5) for 15 days. Group 2 had significantly higher amounts of MPs in the stomach, gills, and feces, electrolyte imbalances (HCO3, Fe, Na+, K+, Ca+2, Cl-, and anion gap, hematobiochemical alterations, and decreases in the activities of superoxide dismutase, catalase, total antioxidant capacity, and glutathione S-transferases compared to the control group. Additionally, Group 2 had significant increase in the percentage of poikilocytosis, and nuclear abnormalities in RBC's compared to the control group. The co-treatment of MPs-exposed fish with lycopene, citric acid, and chlorella-supplemented diets ameliorated the hematological, biochemical, and erythron profile alterations, but only slightly enhanced the antioxidant activity. Overall, lycopene, citric acid, and chlorella can be recommended as a feed supplement to improve hematobiochemical alterations and oxidative damage induced by MPs toxicity in the African catfish (C. gariepinus).